Motorists whose eyesight falls below the legal limit are ignoring advise and continuing to drive, a report has found.
Almost half of optometrists in the UK have seen a patient in the last month who continues to drive despite being told their vision is not good enough, according to a survey by the Association of Optometrists (AOP).
Currently, an estimated 2,900 injuries on our roads each year are caused by drivers with poor vision, according to analysis by RSA Insurance Group.
The law in the UK states that drivers must take a sight examination, which involves reading a number plate from 20 metres away, when they first take their driving test.
From then on motorists self-declare that their eyesight is good enough when they renew their license, and there is no legal requirement to have further eye tests.
The AOP says these laws are too lax, and is calling on the Government to introduce mandatory eye tests for drivers at least every ten years.
Gillian Jones, whose father Ambrose Skingle was killed in 2012 by a motorist who lied about their eyesight when renewing their driving licence, backed the calls to make eye tests a legal requirement.
Mr Skingle, 86, was hit by a motorist who had been told not to drive by an optometrist.
Ms Jones said: “I have two sons and my father was a big part of their lives. Dad taught them how to ride bikes and play golf.
“We had a family dinner together every Sunday. Life has never been the same. It was as if centre of our lives had been ripped out.
“I know some people don’t want to have a sight test because they don’t want the bad news that they have to stop driving.
“I’d like them to think of the consequences, both to themselves and to others. I think most people would feel awful knowing they were responsible for taking a life.
“People have got to look at the bigger picture.”
The AOP has now launched a campaign called ‘Don’t Swerve a Sight Test‘ to call for a change in the law, and to encourage drivers to test their vision regularly.
Solicitor Michael Pace, expert in motoring law, said: “I have a long-held belief that people shouldn’t be able to self-certify, because as we get older I think we all reach a point where we shouldn’t be driving.
“It’s clear that some people are self-certifying who shouldn’t be.
“I’ve dealt with an increasing amount of older drivers who are being stopped by police for careless driving, sometimes because of a lack of confidence and sometimes because of poor eyesight.
“It’s time that we move towards some sort of compulsory eye test.”


Published every weekday, the Switchboard highlights five tech policy stories you need to read.
The FCC is accusing this company of cheating the deaf. "The Federal Communications Commission announced Friday that it will issue a $11.9 million fine against Purple Communications, for allegedly improperly billing a government fund dedicated to providing assistive communication services to the deaf, hearing-impaired and those with speech impairments," writes the Switch's Hayley Tsukayama.
White House seeks legal immunity for firms that hand over customer data. "The White House has asked legislators crafting competing reforms of the National Security Agency to provide legal immunity for telecommunications firms that provide the government with customer data," Spencer Ackerman at The Guardian reports.
When hitting ‘Find My iPhone’ takes you to a thief’s doorstep. "With smartphone theft rampant, apps like Find My iPhone offer a new option for those desperate to recover their devices, allowing victims like Ms. Maguire to act when the police will not," writes Ian Lovett for the New York Times. "But the emergence of this kind of do-it-yourself justice — an unintended result of the proliferation of GPS tracking apps — has stirred worries among law enforcement officials that people are putting themselves in danger, taking disproportionate risks for the sake of an easily replaced item."
Drones banned at Yosemite National Park. "The National Park Service is warning visitors to Yosemite National Park that drones 'are prohibited within park boundaries,'" reports David Kravets at Ars Technica. "The service announced the decision Friday and cited a federal law that says 'delivering or retrieving a person or object by parachute, helicopter, or other airborne means, except in emergencies involving public safety or serious property loss, or pursuant to the terms and conditions of a permit' is illegal."
How Biden helped Hollywood unlock China. "During a luncheon two years ago in Los Angeles, Vice President Biden convinced China's vice president to agree to a deal that would unlock new fortunes for Hollywood," reports the Post's Cecilia Kang. "Biden asked Xi Jinping to relax China's quota of allowing only 20 foreign films to be shown at a time and to increase distribution fees for Hollywood firms."


text:
Bosnia & Herzegovina – This week saw the news that BHRT, Bosnia’s state television station, will temporarily stop broadcasting from 30th June due to its financial situation. The EBU has today said that it is deeply concerned and that this would be the first time a public service broadcaster in Europe would be forced to stop broadcasting because of financial difficulties.
The financial situation of BHRT has been deteriorating over many years with an insufficient level of funding caused by the progressive erosion of the licence fee, a weak collection system and substantial payment arrears from regional partner broadcasters Radio-televizije Federacije BiH and Radio-televizije Republike Srpske. Unfortunately, the necessary funding reforms for public service media in Bosnia and Herzegovina have been repeatedly deferred.
The EBU statement revealed that it has been actively supporting BHRT to find secure funding over recent years. It has met on several occasions with government representatives to try and come up with a long-term solution for the licence fee and has called upon the European Parliament and the European Commission to support the implementation of EU standards in the media of the country.
The EBU has revealed that BHRT owes over CHF 6 million. The BHRT board of directors cited the debts that Bosnia’s two entities’ television networks, RTRS (Radio-Television Republika Srpska) and FTV (Radio-Television of the Federation of Bosnia and Herzegovina), have built up over the years, because they didn’t transfer to BHRT the revenues from the some of the taxes they collect and commercials they sell, despite being legally obliged to do so.
“The total debt that the two entities’ networks have with BHRT is 12.77 million euros: Radio-Television FBiH 8.4 million euros, and Radio-Television RS 4.49 million euros,” the board of directors said.
BHRT was unable to fulfill its financial obligations including payment for the rights to the forthcoming European Football Championships. Several unsuccessful attempts have been made to establish a payment plan for the debt. The initial deadline of 31st May has been extended until 8th June with the hope that the country’s government will take action. BHRT has repeatedly asked Bosnian Council of Ministers in recent months to approve a reform of the law on the public radio-television system, in order to solve the long-standing problems of its financing.
However, so far the Bosnian Parliament was only able to approve a temporary extension of the current system until June 30.
The situation in Bosnia mirrors that in Romania, which resulted in Ovidiu Anton being barred from performing in Stockholm due to TVR’s unpaid debts to the EBU.
BHRT has previously missed Eurovision Song Contest due to financial problems between 2013 to 2015. This year’s entry, Dalal & Deen feat. Ana Rucner and Jala, was paid for by the entry’s own production company and several sponsors, but did not make it out of the semi-final.

domain: news


text:
In the United States, one of the companies, which work together with the government on healthcare plans, is Innovacare, Inc. Innovacare is an innovative healthcare organization whose corporate office is in New Jersey. The company has two other subsidiaries namely MMM Healthcare, Inc. and PMC Medicare Choice, Inc., which operate in Puerto Rico. Through these companies, Innovacare, Inc. has made a tremendous effort to improve healthcare in Puerto Rico. Since the destruction of Puerto Rico by Hurricane Maria, the island has been in need of extensive medical care, and that is why InnovaCare has worked round the clock on the same.
Through the Government’s Health Plan that involves innovative medical technology and therapeutic services to patients, InnovaCare Health together with other providers has made the plan successful. Regarding this, InnovaCare Health rated the highest provider under the Medicaid and Medicare programs. Additionally, Puerto Rico government awarded the company with the NCQA accreditation. Innovacare is the only health care provider to achieve such an award. The company has reached this level through the leadership of Mr. Richard Shinto who is the Chief Executive Officer and the President of the company and Ms. Penelope Kokkinides, the Chief Administrative Officer. The two are well-recognized people with high experience in managerial and medical fields.
Having worked under the Medicaid and Medicare programs, Ms. Kokkinides has boosted the collaboration of Innovacare Health and the government through her experience hence the success of the health plan, reveals Bloomberg. Ms. Kokkinides has worked at Centerlight Health Care as the Vice President and Touchstones Health as the Chief Operating Officer. At Americhoice’s Care and Disease Management docket, she has worked as the Corporate Vice President. Dr. Rich Shinto, who is a highly respected person in medical care docket, on the other hand, is not new in the healthcare field.
At Aveta, Inc., he worked as the CEO of the MMM Healthcare Plan. Other areas he has worked include Cal Optima Health Plan, Medical Pathways Management Company and Med Partners. He has reformed the InnonavaCare Health’s payment system that has shifted from quantity to quality. The combination of managerial experience of Ms. Penelope Kokkinides and Dr. Rich Shinto has indeed helped Innovacare, Inc. to offer the desired medical help to residents of Puerto Rico. Dr. Richard Shinto and Ms. Penelope Kokkinides lead Innovacare Health to improve Puerto Rico’s healthcare program. Their input will never be forgotten and will have a long lasting impact.
Check Penelope Kokkinides out profile on linkedin.com.


text:
The Community Safety Partnership is a multi-agency group comprised of representatives from the police, council, fire service, probation and voluntary sector. Together with members of the community they all work together to make Eastbourne an even safer place.
The Eastbourne Community Safety Partnership, which includes the police, the council and registered housing providers, works together to tackle anti-social behaviour and hate incidents.
Eastbourne’s Prevent programme sets out to achieve this through a multi-agency approach. Working effectively alongside communities to provide interventions and safeguarding measures. Our programme includes safeguarding, staff training and community partnership work.
Prevent is a key part of the Governments counter-terrorism strategy. It aims to divert vulnerable people away from supporting or being drawn into terrorist activity.
Oliver Jones (Strategy and Partnership Lead (Housing and Community)
(Bill & Warwick organise the meetings)

domain: sociology


text:
How Long Do Desert Roses Bloom?
No one looks her best in bad lighting, and the desert rose (Adenium obesum) is no exception. Without enough sunlight during the growing season, this desert dwelling succulent will flower half-heartedly. Grow desert rose in a container outdoors in summer and indoors in the winter, or if you live in U.S. Department of Agriculture plant hardiness zones 11 through 12, you can plant desert roses out in the garden according to University of Florida IFAS Extension.
Adenium Flowering Season
The Adenium flowering season begins with a few flowers in late winter followed by new foliage and continued flowering through summer and fall. The 2- to 3-inch-diameter flowers bloom in white, pink, purple and red which last about seven days. The frequency and profusion of Adenium obesum flowers varies greatly between cultivars and individual plants according to Arizona-Sonora Desert Museum. Desert rose benefits from a dormancy period in winter to allow the plant to rest and recharge before blooming again.
Put desert rose outdoors in the spring when the night temperatures stay above 55 F. If any chance of a late freeze exists, bring desert rose back indoors because this frost-sensitive plant suffers damage below 35 F. To continue blooming, desert rose needs a minimum of six hours of bright sun, preferably filtered in the afternoon through a dappled canopy or shade cloth. You can grow desert rose indoors through the summer in a bright room or on a sun porch but note these important Adenium flowering tips: desert rose will not flower in low light or low temperature conditions.
While desert rose does not tolerate cold at all, it can take the heat. The ideal temperature for growth and blooming falls between 75 and 95 F, but temperatures in triple digits don't cause any harm. Your desert rose can survive outdoors up to 120 F, but summer temperatures above 95 F cause a short dormancy during which the plant will stop blooming. Expect the Adenium flowering season to resume when temperatures return to the optimum temperature range.
Adenium Care Tips
Feed desert rose twice a month with balanced fertilizer but avoid getting the leaves or flowers wet. Start fertilizing in late winter when you see the first buds and continue to fertilize during the summer to support blooming. Rootbound plants grow slowly so repot to encourage growth. You can cut out the top of the stem to encourage branching but take care not to touch or ingest the milky white sap that broken branches excrete as the Adenium obesum latex is toxic.
Water when the potting soil dries out, once a week or more in hot weather. Water occasionally through the winter but allow the potting soil to dry out in between watering and stop watering if the plant goes completely dormant. Use a soil mix that drains easily to avoid root rot. While generally resistant to pests or disease, Adenium care includes checking for fungal leaf spot, mealybugs, scale and aphids.
Move the container into a low-light area and pause your fertilizing schedule in late fall. Keep desert rose between 35 F and 55 F to allow the natural dormancy period to set in. While allowing a dormancy period is recommended, you can prevent dormancy and encourage foliage and flowers through the winter by keeping desert rose in a bright area, maintaining a minimum temperature of 75 F and continuing the regular fertilizer and watering schedule. Disrupting the normal cycle of growth and dormancy, however, may shorten the Adenium lifespan.

domain: agronomy


Victory in Tyra Hunter case 12/11/98
GLAA to Corporation Counsel: Stop Undermining Human Rights Act
[Note: The following memo was hand-delivered to DC Corporation Counsel Robert Rigsby at a meeting he held with representatives of GLAA on Wednesday, June 7. The meeting, one of a series of meetings that Rigsby has held with GLAA, was cordial and productive. This memo cites evidence that Corporation Counsel attorneys have repeatedly argued in court that citizens do not have the right to sue the DC Government for discrimination under the DC Human Rights Act. Rigsby immediately pledged to investigate the matter and report back to GLAA. This problem came to the attention of GLAA as a result of the Tyra Hunter case, in which the government has made the same argument an argument which has been repeatedly rejected by judges.]
Wednesday, June 7, 2000
|Public Safety Chair
|Gay and Lesbian Activists Alliance of Washington, DC
|The right of individuals to sue the DC Government
|under the Human Rights Act
After the Hunter case, which still has not been settled, your office has continued to argue in court that individuals who claim that they have been victims of discrimination by the government are prohibited from suing the District or any DC agency or employee under the DC Human Rights Act. This has occurred in two separate cases, which I will discuss separately.
1. In Jefferson v. DC, et al., in which a Fire Department EMT referred to a 6-year-old asthmatic struggling for breath as a "nigger child," refused to touch the child and delayed emergency transport to Children's Hospital, Corporation Counsel attorneys made this outrageous and discredited argument not once but twice before Judge Jose Lopez. They lost once when the Judge ruled against their motion to dismiss. Unfazed, they questioned the Judge's decision and reargued their position. In rejecting their renewed argument, Judge Lopez wrote the following in his January 11, 2000 ruling on the motion:
Defendant's [District of Columbia] first argument, as was made in their motion to dismiss, is that Plaintiff has failed to exhaust her administrative remedies as required under section 1-2543 of the D.C. Code. Defendant DC believes that under this statutory provision, the Mayor "has authority to render decisions under [the] HRA for both District employees and private citizens." After a thorough search, this court was unable to locate any case law that clearly defines the scope of this section, with respect to whether it is applicable to both private individuals and employees of the District of Columbia. There are, however, Court of Appeals decisions that are very instructive on the scope of this statutory requirement.
In Williams v. District of Columbia, an employee of the University of the District of Columbia initially filed a complaint with the Office of Human Rights claiming employment discrimination on the basis of sex and race. 467 A.2d 140 (D.C. 1983). Unable to settle the dispute, the Plaintiff voluntarily withdrew her complaint and filed suit in the Superior Court. Id. at 141. In affirming the dismissal of Plaintiff's complaint, the Court of Appeals found that "the administrative remedies provided by D.C. Code section 1-2543 and its predecessors are the exclusive remedies available to a District of Columbia government employee claiming [employment] discrimination." Id. at 142. The court went on to hold that "the private right of action established in D.C. Code section 1-2556 and its predecessor, is the only available to non-government employees." Id. The decision in Williams was later re-affirmed by the Court of Appeals in Kennedy v. District of Columbia, 654 A.2d 847 (D.C. 1995). The Court in Kennedy held that "[i]n Williams we clearly stated that D.C. government employees, unlike non-government employees, are required to exhaust the administrative remedies available to them under the D.C. Human Rights Act." Id. at 863. See also Newman v. District of Columbia, 518 A.2d 689, 701 (D.C. 1986) (stating that "[i]n Williams, we identified the administrative exhaustion requirement for government employees in D.C. code section 1-2543").
From these decisions, it is evident that section 1-2543 of the D.C. Code, which requires the exhaustion of administrative remedies, only applies to claims made by District of Columbia employees, not by private individuals. In contrast, section 1-2556 applies to all other individuals who seek redress under the D.C. Human Rights Act. There has been no indication by the Court of Appeals, as Defendant asks this court to conclude, that all individuals filing claims under the D.C. Human Rights Act need to exhaust internal administrative remedies before proceeding to the Superior Court. Only, as Plaintiff correctly points out, those individuals employed by the District of Columbia at the time of the incident need comply with the requirement of exhausting all administrative remedies first. As such, it seems clear to this court that there would be no need for Plaintiff in the instant matter to seek the decision of the Mayor with respect to what steps needed to be satisfied to pursue a discrimination claim. Plaintiff has properly complied with the statutory requirements with respect to her Human Rights Act claim and this court, therefore, has jurisdiction over the instant case.
Jefferson v. District of Columbia et al. , slip. op. at 3-4 (January 11, 2000).
2. On March 27, 2000, in a memorandum opinion in the case of Freddy Ramirez et al. v. District of Columbia, U.S. District Judge Thomas F. Hogan expressly rejected the Corporation Counsel's exhaustion-of-administrative-remedies defense to the Plaintiff's DCHRA claim. The case was brought by Freddy Ramirez, a disabled D.C. elementary school student whose wheelchair didn't fit in the bathroom door, and who as a result had to drag himself into the bathroom in order to relieve himself. The Corporation Counsel, as in Hunter and Jefferson, argued that private citizens cannot sue the D.C. Government. Judge Hogan ruled that the restriction on direct suits applies only to D.C. Government employees. Judge Hogan wrote the following:
DCPS [District of Columbia Public Schools] argues that Freddy [Ramirez] is not entitled to relief under the DCHRA, because he (1) failed to exhaust his administrative remedies before filing suit and (2) failed to prove discriminatory intent as part of his cause of action. However, the statutory exhaustion requirement does not apply to private litigants, such as Freddy, and discriminatory intent is not required if the practice has a discriminatory effect. Therefore, because Freddy has established that DCPS is liable for discrimination, he is entitled to relief under the DCHRA.
The DCHRA provides that a private right of action "in any court of competent jurisdiction" is afforded "any person claiming to be aggrieved by an unlawful discriminatory practice.
In general, the DCHRA does not require a plaintiff to exhaust his administrative remedies, but there is a statutory exhaustion requirement for D.C. government employees. See D.C. Code § 1-2543; See also Hunt v. District of Columbia Dept. of Corr.'s, 41 F. Supp. 2d 31, 37 (D.D.C. 1999) (citing Newman v. District of Columbia, 518 A.2d 698, 700 (D.C. 1986)). However, because Freddy is not an employee of the District, § 1-2543 is inapplicable, and he is entitled to the cause of action set forth in § 1-2556.
Ramirez et al. v. District of Columbia, Memorandum Opinion, at 7 (March 28, 2000).
In summary: Your office continues to make these legally erroneous arguments in order to deny individuals their day in court to determine whether they have a legitimate claim under D.C.'s comprehensive Human Rights Act. We are not suggesting that the government should simply concede every case in which an individual sues the government for discrimination. Of course we expect the government to defend itself. However, it is an abuse not only of the Corporation Counsel's responsibility to follow the law, but of the people who elect this government, to argue that no one in the District may bring suit against the government under the D.C. Human Rights Act. As the largest service provider in the city, the District government has both the power and the greatest potential to violate the civil rights of a citizen of Washington. This was understood by the framers of the DC Human Rights Act, and has been understood by every judge who has ruled on the issue. It is time for the Corporation Counsel to stop trying to hold the government above the Human Rights Act. Please order your staff attorneys to desist from making this argument, and to withdraw the argument in cases where they have made it.
Thank you for your attention to this.


text:
Acetyl-l-carnitine (ALCAR) improves the symptoms of peripheral neuropathy and promotes nerve regeneration, according to a British study published in the July 23rd edition of AIDS.
Distal symmetrical polyneuropathy (DSP), a type of peripheral neuropathy, is a side effect of certain nucleoside analogue reverse transcriptase inhibitors (NRTIs), especially the dideoxynucleotide drugs d4T (stavudine or Zerit), ddC (zalcitabine or Hivid), and ddI (didanosine or Videx). Toxic neuropathy affects 11-66% of patients taking NRTIs. Given the lack of effective therapies for DSP, discontinuation of NRTIs is often necessary to relieve neuropathic pain.
NRTI-associated neuropathy is thought to result from the disruption of mitochondrial DNA synthesis in nerve cells. When this happens, mitochondrial metabolism in the neurones is decreased and the cells’ long peripheral axons die back, leading to reduced innervation (presence of nerve fibres) in the skin and causing pain, tingling, and numbness in the feet and hands.
Noting that ALCAR is vital for normal mitochondrial metabolism and that serum ALCAR levels are decreased in patients with NRTI-associated neuropathy, Dr. Mike Youle, from the Royal Free Centre for HIV Medicine and colleagues conducted a study of the efficacy of ALCAR supplements in improving neuropathic symptoms. Past research has shown that ALCAR promotes the action of nerve growth factor (NGF), stimulates peripheral nerve regeneration, alleviates pain in patients with diabetic neuropathy, and provides short-term relief of NRTI-associated neuropathy symptoms.
The present study included 21 participants with NRTI-associated DSP who received 1500 mg ALCAR twice daily; the trial also included five HIV-negative control subjects without neuropathy. At study entry the median age was 40 years. About 40% had an HIV viral load below 400 copies/mL and the median CD4 count was 286 cells/mm3. Four subjects had asymptomatic HIV disease, eight had symptomatic disease, and eight had frank AIDS. Eight subjects started the study with grade 1 neuropathy (requiring no pain medication), 10 had grade 2 (requiring non-opioid analgesics), and three had grade 3 (requiring opiates).
Skin biopsies were taken from the lower leg before the start of ALCAR therapy and every 6-12 months thereafter (control subjects had only one biopsy). Samples were “immunostained” using various antibodies and examined under a fluorescent microscope to assess the type and quantity (total area) of nerve fibres in the epidermis (outer skin layer), dermis (inner skin layer), and sweat glands.
Subjects were followed for a mean of 14 months (range 5-33 months). In the pre-treatment biopsies, the DSP patients had reduced nerve fibres in the epidermis, the subdermal plexus, and around the sweat glands compared to healthy control subjects. After six months of ALCAR therapy, nerve fibre morphology normalized.
Looking at all nerve fibre types (identified by protein gene product [PGP] antibody staining), epidermis innervation increased by 34% in the DSP patients after six months of ALCAR therapy, and by 101% after 12 months. Dermis innervation increased by 65% after six months, and exceeded that seen in the control subjects by 24 months. Sweat gland innervation increased by 75% after six months and thereafter remained stable (p
Overall, after six months of ALCAR, nerve fibre density in the DSP patients reached the range found in normal skin (92% of the level of epidermis innervation seen in the control subjects, 80% of the dermis innervation, and 69% for sweat glands). After 24 months of ALCAR, improvement continued (epidermis and dermis) or stabilised (sweat glands).
In addition, the grade of neuropathic pain improved in 76% of ALCAR-treated patients and remained unchanged in 19%. Seven patients improved from grade 1 to grade 0 (asymptomatic), four went from grade 2 to grade 0, and one went from grade 3 to grade 0. Those who stopped ALCAR experienced a worsening of neuropathy symptoms. ALCAR was safe and well tolerated in this study, and was not associated with any adverse side effects.
The mechanism by which ALCAR counteracts NRTI toxicity is not known, but it may reduce mitochondrial DNA damage by a direct antioxidant effect, or it may promote the metabolism of fatty acids and glucose. The researchers speculated that ALCAR may offer benefits besides improving peripheral neuropathy, since mitochondrial toxicity also appears to play a role in lipodystrophy syndrome associated with antiretroviral therapy.
“ALCAR treatment improves symptoms [and] causes peripheral nerve regeneration,” the authors concluded. “ALCAR may now offer an effective, pathogenesis-based management approach, allowing patients to remain on NRTI therapy.”
Hart AM et al. Acetyl-l-carnitine: a pathogenesis based treatment for HIV-associated antiretroviral toxic neuropathy. AIDS 18: 1549-1560, 2004.

domain: medical


text:
Drinking it in
Pat Dowding and Sarah Gaunt, Pegram Walters, analyses some of the factors that separate the girls from the boys in the bar, and suggest some improvements to drinks marketing practice
In today's market, women play a strong role in both the purchase and consumption of alcohol, and female brand approval for the drinks consumed by men can be a driving factor in a brand's success.
While men and women have extended their repertoire of drinking in and out of the home, the alcohol market overall has been relatively static in total volume terms. However, the last ten years have seen some significant shifts in volume and some major changes in consumer profile and behaviour in certain sectors, largely driven by social change. It is a fascinating market which is still coming to terms with the power women exert.


text:
VIEWS: 4 PAGES: 11 POSTED ON: 10/15/2012
Cytokines, key players to turn on/off the anti-Trypanosoma cruzi innate defense mechanisms Eugenio A. Carrera-Silva1, Susana Gea2 and Natalia Guiñazú3* 1 Department of immunobiology, School of Medicine, Yale University. New Haven, USA. 2 CIBICI-CONICET, Universidad Nacional de Córdoba. Haya de la Torre y Medina Allende s/n, 5000. Córdoba, Argentina. 3* IDEPA-CONICET, Universidad Nacional del Comahue. Buenos Aires 1400, 8300. Neuquén, Argentina. The early host resistance against Trypanosoma cruzi infection depends on a complex interplay among cytokines, chemical mediators and cells. The major innate immune mechanism against intracellular parasites in phagocytes relies on the production of reactive oxygen species (ROS) and reactive nitrogen species (RNS). The initial step for ROS production is the generation of superoxide anion catalyzed by the enzyme NADPH oxidase. The phagocyte´s NADPH oxidase is a multiprotein complex, which exists in the dissociated state in resting cells and assembles into the functional complex upon stimulation. Additionally, the high amount production of the RNS nitric oxide (NO) depends on the enzyme nitric oxide synthase induction by cytokines. The combination of superoxide anion and NO yields peroxynitrite, the most parasite- harmful reactive species. The anti-inflammatory and pro-inflammatory cytokine balance modulates the activation and induction of both enzymes. Here we discuss the cellular processes involved in macrophage-mediated host defense against Trypanosoma cruzi, and the implications of ROS, RNS and cytokine regulation in host resistance. Keywords Cytokines, NADPH oxidase, Nitric oxide synthase, Phagocyte, Trypanosoma cruzi. 1. Activation of innate immune response and microbicidal mechanisms The adequate activation of innate immune response is essential to control the invading pathogens and to maintain homeostasis. In the past decade, there have been important advances in our understanding of how the inflammatory response to pathogens is initiated. Thus, a typical inflammatory response consists of four components: inflammatory inducers, which initiate the inflammatory response and are detected by sensors. Sensors of innate immunity, such as Toll-like receptors (TLRs), are expressed on specialized sentinel cells including tissue-resident macrophages, dendritic cells, and mast cells. When innate immune receptors are triggered, they induce the production of mediators, including cytokines, chemokines, bioactive amines, eicosanoids, and products of proteolytic cascades, such as bradykinin. These inflammatory mediators act on various target tissues to elicit changes in their functional states that optimize adaptation to the noxious condition. The acute inflammatory response is normally terminated once the triggering insult is eliminated, the infection is cleared, and damaged tissue is repaired . The type of response induced under a given condition depends on the nature of the inflammatory trigger. For example, bacterial pathogens are detected by tissue-resident macrophages and induce the production of inflammatory cytokines (e.g., TNF, IL-1, IL-6) and chemokines (e.g., CCL2 and CXCL8), as well as prostaglandins. Viral infections induce the production of type-I interferons (IFNα, IFNβ) by infected cells and the activation of cytotoxic lymphocytes, whereas infections with parasitic worms lead to the production of histamine, IL-4, IL-5, and IL-13 by mast cells and basophils. Cytokines are fundamental to induce microbicidal mechanisms against pathogens and to drive the adaptive immune response as well [1- 3]. Macrophage role is pleiotropic and include antigen presentation, target cell killing, regulation of inflammatory response, removal of cell debris and promotion tissue remodeling. These functional responses are dependent on micro- environmental signals. A key immunological function of phagocytes is to control invading microorganisms. This property lies on the capacity to generate high amounts of reactive oxygen species (ROS) and reactive nitrogen species (RNS). ROS are oxygen-derived small molecules, such as superoxide anion (O2-), hydroxyl radical and anion (OH and OH-), hydrogen peroxide (H2O2) and hypochlorous acid (HOCl), while RNS are nitrogen-containing oxidants such as nitric oxide (NO) and peroxynitrite anion (ONOO-). ROS and RNS are unstable and rapidly interact with a large number of target molecules including proteins, lipids, carbohydrates, and nucleic acids. ROS and RNS through such interactions may irreversibly destroy or alter the function of the targeted molecule . This chapter will discuss the activation mechanisms of some enzymes responsible of ROS and RNS generation, the contribution of anti- and pro-inflammatory cytokines. The main microbicidal mechanisms involved in the control of Trypanosoma cruzi acute infection will be reviewed. 1.1 NADPH oxidase Phagocytes such as polymorphonuclear neutrophils (PMN), eosinophils, monocytes and macrophages constitute one of the most powerful means of host defense against microbes . Upon infection and inflammation, leukocytes migrate towards the infection site attracted by chemoattractants such as the complement fraction C5a, the N-formyl-methionyl- leucylphenylalanine (fMLF) peptide, IL-8, platelet activating factor (PAF) or leukotriene B4 (LTB4). At the infection site, phagocytes recognize and engulf the microbe. After internalization, phagosomes subsequently fuse with intracellular granules to form the phagolysosome, within which microbial killing is achieved by a combination of non- oxidative and oxidative mechanisms. Potent non-oxidative killing mechanisms include antimicrobial peptides (cationic proteins, defensins, lysozyme, etc). The oxygen-dependent killing mechanism involves the generation of ROS. Together these different mediators contribute to the elimination of the internalized microorganism . ROS are produced by phagocytes in a potent "oxidative burst", characterized by a rapid increase in oxygen uptake, an increase in glucose consumption and abrupt ROS release. The enzyme responsible for superoxide anion (O 2-) production is the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. NADPH oxidase enzyme uses electrons derived from intracellular NADPH to generate O 2-. Much of the O2- produced is dismuted by superoxide dismutase to produce oxygen and hydrogen peroxide (H2O2) (Figure 1), which within neutrophils, is converted by myeloperoxidase to hypochlorous acid and chloramines inside the phagosome. All the products of the oxidative burst are part of the strong microbicidal system of phagocytes . e H2O2 xid e ero as Fig. 1 Schematic representation of ROS formation from up mut S is Hydrogen D peroxide superoxide anion. NADPH oxidase complex reduces oxygen (O2) and generate superoxide anion (O2-) using NADPH as an NADPH electron donor. Much of the O2-- is subsequently transformed oxidase Iron O2 O2- OH- + OH into other ROS. By superoxide dismutase to produce hydrogen peroxide (H2O2), in the presence of iron H2O2 is Oxygen Superoxide Hydroxyl anion transformed to hydroxyl anion and radical (OH- and OH) via anion Ni and radical tric the Fenton reaction and the Haber-Weiss reaction. oxi de OONO- Superoxide anion in the presence of nitric oxide (NO) reacts to yield peroxynitrite anion (OONO-). Nitric oxide Peroxynitrite synthase On the other hand, it has been reported that O2- is also produced in a NADPH-dependent manner in non-phagocytes cells. Because gp91phox (NOX2) is not expressed in these cells, the presence of non-phagocyte-type NOX2 homologs were found in virtually all tissues and have been implicated in many areas of biology, including cell growth, apoptosis, and cancer, innate immunity and inﬂammation, angiogenesis and blood pressure regulation, cell signaling and motility, and hormone synthesis. To date, five NOX isoforms (NOX1, NOX2, NOX3, NOX4, and NOX5) and two related enzymes, DUOX1 and DUOX2, were reported. The NOX family members are now recognized as producers of low levels of ROS that play critical roles in maintaining normal physiologic processes and in the etiology of multiple diseases . NADPH oxidase is a multicomponent enzyme that in leukocytes is comprised by the cell-membrane-bound gp91phox (NOX2) and p22phox proteins (which together constitute the flavocytochrome b558), and the cytoplasmic proteins p47phox, p67phox, p40phox, and Rac. NADPH oxidase is dormant in resting cells but becomes active when cells are stimulated (Figure 2). 2O2 2O2- Extracellular space Extracellular space H+ or phagosome or phagosome gp91 p22 gp91 p22 Intracellular Intracellular Rac GTP p47 space space p67 Rac GDP p40 Rho GDI p47 NADPH NADP+ p67 p40 Fig. 2 Schematic representation of resting and activated phagocyte NADPH oxidase. The proteins gp91phox and p22phox are associated at the plasma membrane and stabilize each other. NADPH oxidase activation is regulated by the organizer p47phox and activator p67phox subunits, and requires GTP-bound Rac. The cytosolic subunits p47phox, p67phox and p40phox are preassembled in the cytosol and upon activation translocate to the membrane, where gp91phox and p22phox are located. In resting cells, Rac is found in a GDP-bound state stabilized by RhoGDI. With stimulation, Rac translocates to the membrane independently of p47phox and p67phox. GTP-bound Rac interacts with p67phox. After the phagocyte cell is activated through the action of soluble chemoattractants and chemokines, or phagozitable particles, the stimulated NADPH oxidase enzyme catalyzes the one-electron reduction of oxygen to produce superoxide anion by using NADPH as substrate. This enzyme uses electrons derived from intracellular NADPH to generate superoxide anion, which subsequently is transformed into other ROS. In the cytoplasm of resting cells are two protein complexes, one composed of p47phox, p67phox and p40phox and the other containing Rac and RhoGDI, the GDP dissociation inhibitor for Rho. Almost 100% of p47phox is located in the cytosol alone or in a complex containing equimolar of p67phox and p40phox [9, 10]. Upon activation, p47phox is heavily phosphorylated, enabling it to co-localize with the flavocytochrome b558 at the membrane . The p47phox protein is believed to be responsible for the transport of the cytosolic subunits to the membrane and is thus considered to be the organizing adaptor protein [12, 13]. On the other hand, p67phox represents an essential activating cofactor, possessing a domain that regulates the reduction of FAD by NADPH . During activation, p67phox and p40phox also become phosphorylated . The significance of p40phox phosphorylation is unknown, although it has been reported to promote negative regulation of NADPH oxidase function by p40phox . The translocation of Rac to the plasma membrane NADPH oxidase in human neutrophils occurs through an independent mechanism from, but in a coordinated manner with, translocation of the p47phox/p67phox complex . NADPH oxidase physiological activator stimuli include phagozitable particles such as bacteria and yeast, certain molecules that induce chemotaxis in these cells, certain bioactive lipids, complement proteins and antibodies. The stimuli of highest importance are phagozytable particles. Coating the surface of a pathogen, by antibodies or complement proteins, to enhance phagocytosis is called opsonization. Antibodies do this in either two ways. In the first, bound antibodies coating the pathogen are recognized by Fc receptors on phagocytic cells that bind to the antibody C- region. Alternatively, antibodies binding to the surface of a pathogen can activate the complement system. Complement proteins binding to the pathogen surface; opsonize the pathogen by binding complement receptors on phagocytes. Phagocytosis is an active process, in which the bound pathogen is internalized in a membrane-bounded vesicle, known as phagosome or endocytic vacuole. The lysosome liberates its content into the phagosome and generates the pahgolysosome. NADPH oxidase enzyme assembles at the phagosomal membrane. Once activated, O 2- production occurs inside of the phagosome. The isolation of superoxide release into the phagosome is important since ROS are able to damage host tissues too. Other non-physiological molecules also activate NADPH oxidase complex and have been important in understanding the regulation of the phagocyte NADPH oxidase. In this sense, PMA and calcium ionophores induce superoxide production, which does not have to be initiated through membrane receptors (PMA activates PKC, calcium ionophores increase the intracellular calcium concentration). 1.2 Nitric oxide synthase Nitric oxide (NO) is a messenger molecule functioning in several processes, such as neurotransmission, vascular regulation and host immune defense, among others. NO is produced by the oxidation of L-arginine by the enzyme NO synthase (NOS). Three NOS isoforms have been reported, neuronal NOS (nNOS), endothelial NOS (eNOS), and inducible NOS (iNOS). Both nNOS and eNOS are constitutive and Ca2+-dependent, while iNOS isoform is calcium- independent and inducible. NOS isoforms not only produce NO, but also a number of species resulting from oxidation, reduction, or adduction of NO in physiological milieus, thereby generating various nitrogen oxides, S-nitrosothiols, peroxynitrite (ONOO-), and transition metal adducts . An important protective role for RNS has been established in macrophage killing of intracellular protozoa [18- 21], bacteria , fungus  and viruses . A similar role has been demonstrated in neutrophil killing of Candida albicans [25, 26] and Staphilococcus aureus . The aminoacid L-arginine is the common substrate of two enzymes NOS and arginase. Arginase enzyme metabolizes L- arginine to produce urea and L-ornithine. In subsequent steps, L-ornithine is metabolized by ornithine decarboxilase (ODC) to produce polyamines (putrescine, spermidine, spermine) and proline, which promote cell growth and collagen production. Arginase exists in 2 isoforms, arginase I enzyme is located in the cytosol and expressed in the liver as one of the enzymes of the urea cycle which detoxiﬁes ammonia in mammals, and arginase II protein is expressed as a mitochondrial protein in a variety of peripheral mammalian tissues, most prominently in kidney, prostate, small intestine and the lactating mammary gland . The reciprocal regulation of iNOS and arginase has been reported in various inﬂammatory pathologies. Arginase activity or expression induction was demonstrated in murine inﬂammatory cell inﬁltrates in schistosomiasis , american trypanosomiasis [30, 32], leishmaniasis [33, 34], viral [35, 36] and bacterial [37, 38] infections. Since the availability of intracellular L-arginine is a rate-limiting factor in NO production, arginase activity may down-regulate NO production by competing with NOS for L-arginine. While in macrophages stimulated by inflammatory Th1 cytokines TNF-α and IFNγ, iNOS is induced, arginase expression in murine macrophages is induced by Th2 cytokines IL-4, IL-10 and IL-13 . In murine disease models that follow the Th1/Th2 paradigm with regard to disease susceptibility or resistance iNOS is induced in the context of a Th1-dominated resistant phenotype while macrophage arginase is up-regulated during Th2-mediated disease progression. In contrast to its involvement in host- detrimental immunopathology, myeloid cell arginase can also serve a crucial host-protective function by down- regulating excessive Th1-induced inﬂammation . 2. Regulation of microbicidal mechanisms by cytokines In the course of infectious diseases, a permanent challenge is imposed on the immune system: to eliminate, or at least to control, the infectious agent and to minimize the destruction of tissue architecture. The extent of the immune response and the action of potentially harmful effector cells should be tightly controlled, either by regulatory cytokines or by the elimination of lymphocytes by apoptosis. Anti-inflammatory response, mediated by IL-10 and TGF, counteract the effects of inflammatory cytokines such as IL-12, TNF and IFN . In this sense, macrophages are innate immune cells with well-established roles in the primary response to pathogens, but also in tissue homeostasis, coordination of the adaptive immune response, inflammation, resolution, and repair. They can function as control switches of the immune system, securing the balance between pro- and anti-inflammatory reactions. For this purpose and depending on the activating stimuli, these cells can develop into different subsets: classically (M1) or alternatively (M2) activated [41- 43]. The classical activation profile occurs in a type I cytokine environment (IFNγ, TNF) or upon recognition of pathogen-associated molecular pattern (PAMPs) such as LPS, lipoproteins, dsRNA, lipoteichoic acid, etc, and endogenous danger signals such as heat shock proteins, etc. As such, they play an important role in protection against intracellular pathogens, and under certain conditions also cancer cells. Classically activated macrophages (M1) typically produce high levels of IL-12 – and in humans also IL-23  – and low levels of IL-10 and are consequently strong promoters of Th1 immune responses. In addition, these cells exert anti-proliferative and cytotoxic activities, resulting partly from their ability to secrete RNS and ROS and inflammatory cytokines (TNF, IL-1, IL-6) [45- 46]. Alternative pathway of macrophage activation may arise in a Th2 cell-biased environment with a stereotypic signature of expressed genes induced by IL-4 and IL-13. As opposed to M1, M2 promote Th2 responses, resulting in effector functions such as parasite killing and encapsulation, but also exacerbation of Th2 cytokine-associated pathologies, such as allergies. These macrophages secrete copious amounts of anti-inflammatory molecules like IL-10 and TGF, thereby down-regulating inflammatory processes initiated by M1. Additionally, M2 exerts selective immunosuppressive functions in T lymphocyte cell proliferation [47, 48]. Their presence in healthy individuals in the placenta, lungs and immune privileged sites, as well as in chronic inflammatory diseases like rheumatoid arthritis, atherosclerosis and psoriasis, further suggests that M2 protect organs and surrounding tissues against detrimental immune responses. Moreover, they increase the fibrinogenic activity of human fibroblasts , promote angiogenesis  and wound repair during the healing phase of acute inflammatory reactions and in chronic inflammatory diseases . The Arginase 1 enzyme is a signature marker intimately associated to M2 and there has been extensive investigation of its possible role in alternative versus classical activation [51- 53]. The cross-regulation of the iNOS-arginase balance by Th1 mediators, such as IFN and LPS, and by Th2 cytokines, such as IL-4 and IL-13, suggests that the measure of NO level and arginase activity in distinct macrophage populations reflects their activation state, classical or alternative. However, the distinction between M1 and M2 solely based on the iNOS-arginase balance could be oversimplified. Other well reported differences between M1 and M2 are situated at the level of cytokine and chemokine secretion, both in human and mouse [54, 55]. NADPH oxidases (NOXs) are activated in response to stimulation of associated receptors, such as those for insulin, platelet-derived growth factor, nerve growth factor, fibroblast growth factor, TNF, the growth factor GM-CSF, and angiotensin. In phagocytes they are activated by engagement of receptors for the antibody Fc chain (Fcγ) or for complement during phagocytosis of opsonized particles. NOX2 in neutrophils is also activated by engagement of receptors for integrins simultaneously with engagement of receptors for soluble agonists such as TNF, formyl peptides, complement C5a, the growth factors G-CSF and GM-CSF, or macrophage inflammatory protein. NOXs can be regulated by phosphorylation of p47phox (or TKS5), by flavinloading of gp91phox, by the GTPase activity of Rac1 or Rac2, and by assembly of their components (gp91phox; p22phox; p47phox (or TKS5); p40phox (or NoxO1); p67phox (or NoxA1); and Rac1 or Rac2) . The priming of NADPH oxidase is a phenomenon caused by certain molecules that induce a weak oxidative response or do not directly activate the NADPH oxidase by themselves, but strongly augment superoxide production triggered by a second stimulus. The priming agents LPS and IFNγ increase the expression level of gp91phox in human monocyte- derived macrophages and neutrophils . Other cytokines such as G-CSF, GM-CSF and TNFα are able to prime NADPH oxidase activity in human PMN. The shared mechanism by which GM-CSF and TNFα prime the PMN respiratory burst involves partial phosphorylation of p47phox . The enhanced phosphorylation of cytosolic components, decreased inhibition of the autoinhibitory region of p47phox, and assembly of p40phox, p47phox and p67phox into complexes in the cytosol before translocation to the membrane have been proposed as possible priming mechanisms . Thus, pro-inflammatory cytokines such as TNF which do not activate NADPH oxidase but prime its activation in response to a secondary stimulus such as fMLF , induce partial phosphorylation of p47phox on Ser345 by ERK1/2 or p38MAPK and promote NADPH oxidase assembly [58, 61]. Although substantial advances have been made to understand the process leading to NADPH oxidase activation, the molecular mechanisms involved in NADPH oxidase inactivation are elusive. Deactivation of NADPH oxidase is critical for timely resolution of inflammation, before bystander tissue damage occurs. Rapid deactivation of NADPH oxidase could be due to several processes, including receptor desensitization, uncoupling of transduction pathways, and modulation of the phosphorylation of NADPH oxidase components such as p47phox and p67phox . Important mediators of this deactivation are also some anti-inflammatory cytokines such as IL-10 and TGF. It has been shown that IL-10 down-regulates PMN ROS production under certain experimental conditions [62-64]. IL-10 is a pleiotropic cytokine that plays a critical role in the regulation of inflammatory responses and immune reactions, acting on both hematopoietic and non-hematopoietic cells . It has been postulated that the inhibition of NADPH oxidase by IL-10, in microglia cells, is mediated through the inhibition of the JAK1 signaling pathway, which in turn prevents the translocation of cytosolic subunit p47phox , in PMN cells it has been reported that IL-10 inhibits p47phox phosphorylation through a decrease in ERK1/2 activity . It has been postulated that the deactivating NADPH oxidase effect of IL-10 could contribute to the tight regulation of the oxidative burst at the inflammatory site. TGF- β1cytokine one of the most potent endogenous immune modulators of inflammation, has also been implicated in NADPH deactivation in microglial cells. The molecular mechanism of its anti-inflammatory effect on the activation of is through the inhibition of NADPH oxidase activity by preventing the ERK-dependent phosphorylation of on p47phox in microglia to reduce oxidase activities induced by LPS . 3. Trypanosoma cruzi infection control by microbicidal mechanisms Trypanosoma cruzi is a flagellated parasite that reproduces asexually by binary fission. Like all other trypanosomes, T. cruzi reside, some stages of their lives, in the blood and/or tissues of vertebrate hosts and during other stages they live in the digestive tracts of invertebrate vectors. In the mammalian host, in the peripheral blood, the trypomastigotes stage is found. Inside the cytoplasm of the nucleated T. cruzi replicates as an amastigote . T. cruzi parasite is the causative agent of Chagas’ disease, and it has been estimated that 10 million people are infected worldwide, mostly in Latin America where Chagas’ disease is endemic. More than 25 million people are at risk of the disease. It is estimated that in 2008 Chagas’ disease killed more than 10 000 people . After acute infection, most patients remain asymptomatic for decades; while a significant fraction of the affected individuals (30%) develop principally heart and gastrointestinal clinical manifestations during chronic Chagas’ disease. It has been postulated that T. cruzi persistence together with the immune response induced to multiple myocardial antigens may participate in the heart damage . T. cruzi parasite actively invades a variety of cell types such as ﬁbroblasts, muscle cells and macrophages, thereby triggering a diversity of molecular interactions that mobilize the host immune response. A broad panel of innate immune cells is the first defense line against T. cruzi, these include neutrophils, tissue macrophages, monocytes, dendritic cells, eosinophils and natural killer cells [71, 72]. Activated phagocytes play a central role in the control of infection through the production of ROS and RNS [73, 74]. It has been well recognized that T. cruzi infection augments the oxidative metabolism of several cell types such as murine spleen cells , murine hepatocytes , blood monocytes and macrophages [77, 78], erythrocytes , and cardiac cells [80, 81]. It was recently demonstrated that the derived parasite antigen cruzipain (Cz- is a parasite glycoprotein member of the papain superfamily, expressed as in all parasite developmental stages) induced ROS production in splenocytes from non-immune and Cz-immune C57BL/6 mice and in a Raw 264.7 macrophage cell line. Cz stimulation favored the production of several ROS, mainly superoxide anion, and also triggered NADPH oxidase activation , this work was the first report unraveling parasite antigens that might be involved in oxidative stress induced by T. cruzi. Besides ROS, it has been reported that T. cruzi infection via inflammatory stimulation induce M1 activated macrophages that produce NO via the activation of iNOS [20, 21]. Experimental evidence certainly implicates RNS as important mediators of parasite killing in mice experimentally infected with T. cruzi [73, 74]. NO has been one of the RNS more frequently studied in the context of T. cruzi infection. In this sense, T. cruzi infection results in elevated NO levels in plasma [83, 84]. Infected mice which were treated with iNOS inhibitors developed high parasitemia and mortality [83, 85, 86]. On the other hand, it has been, reported that iNOS deﬁcient mice infected with the Brazil or Tulahuen strain of T. cruzi are as resistant to infection as wild-type mice. In the absence of iNOS, the infection resistance would be probably mediated by the enhancement of other immune effector mechanisms. Thus, this work pointed out to the importance of other microbicidal mechanism than NO alone . In this sense, Alvarez et al (2011) , have shown that internalization of T. cruzi trypomastigotes by macrophages triggers the assembly of the NADPH oxidase complex to yield superoxide anion. Moreover they demonstrate that the main mechanism responsible of infection control occurred via the production of NO and superoxide anion simultaneously, which generates intraphagosomal peroxynitrites. Taking into account that T. cruzi has a complex life cycle, several parasite derived-antigens might participate in microbicidal mechanisms induction. Glycophosphatidylinositol-anchored mucin-like glycoproteins (GPI)  and Tc52-released antigen  synergize with IFNγ to stimulate NO production by macrophages. Through mice immunization models it has been demonstrated that several parasite derived antigens are able to induce the production of pro-inflammatory cytokines and NO [90- 93]. Currently it remains unclear the host innate receptor that interacts with some parasite derived antigens proven to trigger an inflammatory response. This is not the case for the Tc52- released antigen  and GPI-anchors of mucin like protein  that signal via TLR2. Mice lacking TLR2 display slightly enhanced susceptibility to T. cruzi in vivo and macrophages show impaired pro-inflammatory cytokine production when exposed to the parasite in vitro . In addition, there is evidence demonstrating that Cz in vitro up-regulates TLR2 surface expression on immune total spleen and F4/80+ cells. However, it has not been demonstrated whether Cz is able to interact with TLR2, with further studies required to clarify this issue. Nevertheless, it is probable that Cz modulates functional responses to the T. cruzi ligands that bind TLR2 . Moreover, it has been also demonstrated that Cz is able to modulate the iNOS/arginase balance. The immunization of resistant C57BL/6 mice with Cz primed spleen cells for classical macrophage activation. Cz induced a high level of NO associated with an up-regulation of iNOS protein and messenger expressions in immune cells. These cells also produced high levels of IL-12 and IFNγ, probably as a consequence of this pro-inflammatory milieu spleen cell also demonstrated an amplified NADPH oxidase activation [82, 92]. Altogether these findings indicate that the inflammatory environment induced by Cz in C57BL/6 background may be participating in the priming of immune cells to a higher production of RNS and ROS. By contrast, the immunization of susceptible BALB/c mice with Cz resulted in an enhanced anti- inflammatory (IL-4, IL-5 and IL-10) cytokine secretion, associated with the induction of a CD11b+GR1+ spleen immature myeloid population that exhibited arginase, but not iNOS activity . Furthermore, Cz-stimulated naive macrophages secreted IL-10 and TGFβ and displayed enhanced arginase activity which favored T. cruzi growth [98, 99]. In this sense it has been demonstrated that the induction of ODC activity, an enzyme involved in the synthesis of polyamines, is an essential factors for intracellular parasite replication. These reports indicate the importance of parasite derived antigens, the innate receptor triggered, the cytokine milieu induced as well as the genetic background in the balance parasite resistance/persistence during T. cruzi infection. 4. The cytokine environment favors Trypanosoma cruzi persistence and immune evasion The coordinated interplay between macrophages, neutrophils and monocytes is crucial for the effective elimination of noxious agents and the restoration of tissue homeostasis after injury or infection. Initial phagocyte interactions promote mutual recruitment and antimicrobial activities, but later crosstalk between phagocyte populations dampens pro- inflammatory responses and promotes resolution of the inflammatory response. Although the release of TGFβ, IL-10 and PGE2 can contribute to restoration of homeostasis after tissue injury, these anti-inflammatory mediators also dampen antimicrobial mechanisms. In this context, it has been shown that macrophage-mediated clearance of apoptotic cells promotes intracellular parasite persistence, taken advantage of an immunosuppressive effect. Interestingly, it was reported during parasitic infections by T. cruzi , T. gondii , and L. major , both short range and systemic apoptosis can be induced in lymphoid cells, with impact on cytokine production and parasite evasion from immune responses. Macrophages ingesting apoptotic leukocytes become deactivated, as secretion of the pro-inflammatory cytokine TNF is suppressed by autocrine and paracrine secretion of PGE2 and TGF . Therefore, the phagocytic burial of apoptotic cells could play a detrimental role by deactivating host macrophages, thus facilitating intracellular replication of parasites. Moreover, injection of apoptotic cells increases the parasitemia of T. cruzi infected mice. Freire-de-Lima et al.,  have identified a biochemical pathway initiated in macrophages by contact with apoptotic cells and engagement of V3 integrin. This interaction stimulates PGE2 and TGF production, followed by increased ODC activity, and increased synthesis of the polyamine putrescine [104, 105]. Previous studies demonstrated that the intracellular pathogens are able to inﬂuence the production of various effectors molecules by disabling intracellular signaling pathways required for their synthesis [106- 108]. During T. cruzi infection spleen macrophages from BALB/c mice secreted lower concentration of nitrites than C57BL/6 mouse cells, indicating that impaired function of myeloid cells in susceptible mice could contribute to the persistence of infection . In contrast to the Type 2 response seen in BALB/c, when C57BL/6 mice are immunized with Cz, the induced cytokine profile was predominantly type 1  indicating that the immunization of C57BL/6 mice with Cz primed the spleen cells for classical macrophage activation. Thus, it seems clear that classical activation of macrophages by inflammatory cytokines plays a central role in the control of parasitemia and clearance during acute phase of T. cruzi infection; however the alternative activation favors the replication, persistence and dissemination of the parasite. Recently, in a comparative study during acute T. cruzi infection, it has been demonstrated that hepatic leukocytes from infected B6 mice produced higher amounts of inflammatory cytokines than susceptible BALB/c mice, whereas the anti-inflammatory cytokines IL-10 and TGFβ were only released by hepatic leukocytes from BALB/c. Strikingly, a higher expression of TLR2 and TLR4 was observed in hepatocytes of infected BALB/c mice. However, in infected B6 mice, the strong pro-inflammatory response was associated with a high and sustained expression of TLR9 and iNOS in leukocytes and hepatic tissue respectively. Additionally, in infected B6 mice, co-expression of gp91- and p47- phox NADPH oxidase subunits were detected in liver tissue of infected B6 mice. Notably, the pre-treatment previous to infection with Pam3CSK4, TLR2-agonist, induced a significant reduction of transaminase activity levels and inflammatory foci number in livers of infected B6 mice. Thus, an altered TLR2, TLR4 and TLR9 signaling and exacerbate inflammatory cytokine profile could be responsible of the fatal hepatic damage observed in infected B6. These results demonstrate the importance of adequate TLR signaling on immune and non- immune cells to attenuate the tissue damage during Trypanosoma cruzi acute infection [76, 111]. It has been postulated that T. cruzi molecules, such as glycophosphatidylinositol anchors, may directly initiate IL-12 and NO production, through TLR2 activation, thus promoting host resistance during infection [95, 96]. However, it was also demonstrated that Glycoinositolphospholipids from T. cruzi (GIPL) interfere with macrophages and dendritic cell responses by down regulation of pro-inflammatory cytokines and co-stimulatory markers. Brodskyn C. et al.; 2002 showed that the ceramide portion of GIPL was responsible for most of the activity exhibited by the whole molecule. Considering the important role of the immune response in determining the fate of the host-parasite relationship, the immunoregulatory activities of T. cruzi GIPL are potentially important for parasite evasion and then pathogenesis of infection with protozoan parasites . Innate immunity plays a crucial role to efficiently develop the adaptive immune response. In this context it was freshly reported that insufficient TLR activation contributes to the slow development of CD8+ T cell responses in T. cruzi infection. The authors propose that T. cruzi is a stealth invader, largely avoiding recognition by components of the innate immune system until the infection is well established. This conclusion is supported by the ability to accelerate the induction of T cell responses to T. cruzi by administration of ligands for TLR2 and TLR9 at the time of infection. These studies highlight a previously unappreciated mechanism of immune evasion, the surreptitious establishment of infection, by the protozoan T. cruzi . 5. Concluding remarks The presented data highlight the fact that ROS and RNS are involved in the control of T. cruzi infection. However, the impact of the microbicidal status achieved by phagocytes is not independent of other immune events, such as the levels of inflammatory cytokines as well as second activation signals as PAMPs. Thus, the protective immune response against microorganisms is multifactorial, and NADPH oxidase, ROS, iNOS, NO, and RNS have been significantly implicated. It is noteworthy that during the very early stage of T. cruzi infection the interaction between host macrophage and parasite itself or parasite derived antigens results in the induction of intracellular signaling pathways which also lead to activation or deactivation of the cellular microbicidal mechanisms , which may either inhibit parasite replication or provide a favorable environment to its dissemination. For this reason is particularly important to elucidate the mechanism involved in the defense against pathogens as well as the evasive counterpart, in order to develop successful strategies against infectious diseases. Acknowledgements Financial support received from Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET), Agencia Nacional de Investigaciones Científicas y Técnicas, and Universidad Nacional del Comahue is gratefully acknowledged. S. Gea and N Guiñazú are members of Research Career from CONICET. References  Medzhitov R. Inflammation 2010: new adventures of an old flame. Cell. 2010; 140:771-6  Nathan C., Ding A. Nonresolving Inflammation. Cell. 2010; 140:871-882.  Soehnlein O., Lindbom L. Phagocyte partnership during the onset and resolution of inflammation. Nat Rev Immunol. 2010; 10:427-439  Bedard K, Krause KH. The NOX family of ROS-generating NADPH oxidases: physiology and pathophysiology. Physiol Rev. 2007; 87:245-313.  Nathan C. Neutrophils and immunity: challenges and opportunities. Nat Rev Immunol. 2006; 6:173-82.  Plüddemann A, Mukhopadhyay S, Gordon S. Innate immunity to intracellular pathogens: macrophage receptors and responses to microbial entry. Immunol Rev. 2011; 240:11-24.  Babior BM. NADPH oxidase. Curr Opin Immunol. 2004; 16:42-57.  Bokoch GM. NADPH oxidases in innate immunity. J Innate Immun. 2009 Oct;1(6):507-8.  Park JW, Benna JE, Scott KE, Christensen BL, Chanock SJ, Babior BM. Isolation of a complex of respiratory burst oxidase components from resting neutrophil cytosol. Biochemistry. 1994; 33:2907-11.  Lapouge K, Smith SJ, Groemping Y, Rittinger K. Architecture of the p40-p47-p67phox complex in the resting state of the NADPH oxidase. A central role for p67phox. J Biol Chem. 2002; 277:10121-8.  Ago T, Kuribayashi F, Hiroaki H, Takeya R, Ito T, Kohda D, Sumimoto H. Phosphorylation of p47phox directs phox homology domain from SH3 domain toward phosphoinositides, leading to phagocyte NADPH oxidase activation. Proc Natl Acad Sci U S A. 2003; 100:4474-9.  Dusi S, Donini M, Rossi F. Mechanisms of NADPH oxidase activation: translocation of p40phox, Rac1 and Rac2 from the cytosol to the membranes in human neutrophils lacking p47phox or p67phox. Biochem J. 1996; 314:409-12.  El-Benna J, Dang PM, Gougerot-Pocidalo MA, Marie JC, Braut-Boucher F. p47phox, the phagocyte NADPH oxidase/NOX2 organizer: structure, phosphorylation and implication in diseases. Exp Mol Med. 2009; 41:217-25.  Nisimoto Y, Motalebi S, Han CH, Lambeth JD. The p67(phox) activation domain regulates electron flow from NADPH to flavin in flavocytochrome b(558). J Biol Chem. 1999; 274:22999-3005.  Lopes LR, Dagher MC, Gutierrez A, Young B, Bouin AP, Fuchs A, Babior BM. Phosphorylated p40PHOX as a negative regulator of NADPH oxidase. Biochemistry. 2004; 43:3723-30.  Bokoch GM, Diebold BA. Current molecular models for NADPH oxidase regulation by Rac GTPase. Blood. 2002; 100:2692- 6.  Weigert A, Brüne B. Nitric oxide, apoptosis and macrophage polarization during tumor progression. Nitric Oxide. 2008; 19:95-102.  Liew FY, Li Y, Millott S. Tumor necrosis factor-alpha synergizes with IFN-gamma in mediating killing of Leishmania major through the induction of nitric oxide. J Immunol. 1990; 145:4306-10.  Vincendeau P, Daulouède S. Macrophage cytostatic effect on Trypanosoma musculi involves an L-arginine-dependent mechanism. J Immunol. 1991; 146:4338-43.  Gazzinelli RT, Oswald IP, Hieny S, James SL, Sher A. The microbicidal activity of interferon-gamma-treated macrophages against Trypanosoma cruzi involves an L-arginine-dependent, nitrogen oxide-mediated mechanism inhibitable by interleukin- 10 and transforming growth factor-beta. Eur J Immunol. 1992; 22:2501-6.  Rodrigues MM, Ribeirão M, Boscardin SB. CD4 Th1 but not Th2 clones efficiently activate macrophages to eliminate Trypanosoma cruzi through a nitric oxide dependent mechanism. Immunol Lett. 2000; 73:43-50.  Chan J, Xing Y, Magliozzo RS, Bloom BR. Killing of virulent Mycobacterium tuberculosis by reactive nitrogen intermediates produced by activated murine macrophages. J Exp Med. 1992; 175:1111-22.  Moreira AP, Dias-Melicio LA, Peraçoli MT, Calvi SA, Victoriano de Campos Soares AM. Killing of Paracoccidioides brasiliensis yeast cells by IFN-gamma and TNF-alpha activated murine peritoneal macrophages: evidence of H(2)O (2) and NO effector mechanisms. Mycopathologia. 2008; 166:17-23.  Jarasch N, Martin U, Kamphausen E, Zell R, Wutzler P, Henke A. Interferon-gamma-induced activation of nitric oxide- mediated antiviral activity of macrophages caused by a recombinant coxsackievirus B3. Viral Immunol. 2005;18:355-64.  Fierro IM, Barja-Fidalgo C, Cunha FQ, Ferreira SH. The involvement of nitric oxide in the anti-Candida albicans activity of rat neutrophils. Immunology. 1996; 89:295-300.  Fierro IM, Nascimento-DaSilva V, Arruda MA, Freitas MS, Plotkowski MC, Cunha FQ, Barja-Fidalgo C. Induction of NOS in rat blood PMN in vivo and in vitro: modulation by tyrosine kinase and involvement in bactericidal activity. J Leukoc Biol. 1999; 65:508-14.  Malawista SE, Montgomery RR, van Blaricom G. Evidence for reactive nitrogen intermediates in killing of staphylococci by human neutrophil cytoplasts. A new microbicidal pathway for polymorphonuclear leukocytes. J Clin Invest. 1992; 90:631-6.  Munder M. Arginase: an emerging key player in the mammalian immune system. Br J Pharmacol. 2009; 158:638-51.  Abdallahi OM, Bensalem H, Augier R, Diagana M, De Reggi M, Gharib B. Arginase expression in peritoneal macrophages and increase in circulating polyamine levels in mice infected with Schistosoma mansoni. Cell Mol Life Sci. 2001; 58:1350-7.  Aoki MP, Guiñazú NL, Pellegrini AV, Gotoh T, Masih DT, Gea S. Cruzipain, a major Trypanosoma cruzi antigen, promotes arginase-2 expression and survival of neonatal mouse cardiomyocytes. Am J Physiol Cell Physiol. 2004; 286:C206-12.  Cuervo H, Pineda MA, Aoki MP, Gea S, Fresno M, Gironès N. Inducible nitric oxide synthase and arginase expression in heart tissue during acute Trypanosoma cruzi infection in mice: arginase I is expressed in infiltrating CD68+ macrophages. J Infect Dis. 2008; 197:1772-82.  Stempin C, Giordanengo L, Gea S, Cerbán F. Alternative activation and increase of Trypanosoma cruzi survival in murine macrophages stimulated by cruzipain, a parasite antigen. J Leukoc Biol. 2002; 72:727-34.  Iniesta V, Gómez-Nieto LC, Corraliza I. The inhibition of arginase by N(omega)-hydroxy-l-arginine controls the growth of Leishmania inside macrophages. J Exp Med. 2001; 193:777-84.  Kropf P, Fuentes JM, Fähnrich E, Arpa L, Herath S, Weber V, Soler G, Celada A, Modolell M, Müller I. Arginase and polyamine synthesis are key factors in the regulation of experimental leishmaniasis in vivo. FASEB J. 2005; 8:1000-2.  Mistry SK, Zheng M, Rouse BT, Morris SM Jr. Induction of arginases I and II in cornea during herpes simplex virus infection. Virus Res. 2001; 73:177-82.  Wang HC, Kao YC, Chang TJ, Wong ML. Inhibition of lytic infection of pseudorabies virus by arginine depletion. Biochem Biophys Res Commun. 2005; 334:631-7.  Lewis ND, Asim M, Barry DP, Singh K, de Sablet T, Boucher JL, Gobert AP, Chaturvedi R, Wilson KT. Arginase II restricts host defense to Helicobacter pylori by attenuating inducible nitric oxide synthase translation in macrophages. J Immunol. 2010; 184:2572-82.  Bussière FI, Chaturvedi R, Cheng Y, Gobert AP, Asim M, Blumberg DR, Xu H, Kim PY, Hacker A, Casero RA Jr, Wilson KT. Spermine causes loss of innate immune response to Helicobacter pylori by inhibition of inducible nitric-oxide synthase translation. J Biol Chem. 2005; 280:2409-12.  Peluffo G, Piacenza L, Irigoín F, Alvarez MN, Radi R. L-arginine metabolism during interaction of Trypanosoma cruzi with host cells. Trends Parasitol. 2004; 20:363-9.  Lopes MF, Freire-de-Lima CG, DosReis GA. The macrophage haunted by cell ghosts: a pathogen grows. Immunol Today. 2000; 21:489-94.  Van Ginderachter JA, Movahedi K, Hassanzadeh Ghassabeh G, Meerschaut S, Beschin A, Raes G, De Baetselier P. Classical and alternative activation of mononuclear phagocytes: picking the best of both worlds for tumor promotion. Immunobiology. 2006; 211:487-501.  Gordon S. and Martinez F. Alternative activation of macrophages: mechanism and functions. Immunity. 2010; 32:593-604.  Martinez FO, Helming L, Gordon S. Alternative activation of macrophages: an immunologic functional perspective. Annu Rev Immunol. 2009; 27:451-83.  Verreck FA, de Boer T, Langenberg DM, Hoeve MA, Kramer M, Vaisberg E, Kastelein R, Kolk A, de Waal-Malefyt R, Ottenhoff TH. Human IL-23-producing type 1 macrophages promote but IL-10-producing type 2 macrophages subvert immunity to (myco)bacteria. Proc Natl Acad Sci U S A. 2004; 101:4560-5.  Bonnotte, B., Larmonier, N., Favre, N., Fromentin, A., Moutet, M., Martin, M., Gurbuxani, S., Solary, E., Chauffert, B., Martin, F., 2001. Identification of tumor- infiltrating macrophages as the killers of tumor cells after immunization in a rat model system. J. Immunol. 2001; 167:5077–5083.  Mytar, B., Siedlar, M., Woloszyn, M., Ruggiero, I., Pryjma, J., Zembala, M. Induction of reactive oxygen inter- mediates in human monocytes by tumour cells and their role in spontaneous monocyte cytotoxicity. Br. J. Cancer. 1999; 79:737–743.  Brys, L., Beschin, A., Raes, G., HassanzadBrombacher, F., De Baetselier, P. Reactive oxygen species and 12/15-lipoxygenase contribute to the antiproliferative capacity of alternatively activated myeloid cells elicited during helminth infection. J. Immunol. 2005; 174:6095–6104.  Loke, P., MacDonald, A.S., Robb, A., Maizels, R.M., Allen, J.E. Alternatively activated macrophages induced by nematode infection inhibit proliferation via cell-to-cell contact. Eur. J. Immunol. 2000; 30:2669–2678.  Song, E., Ouyang, N., Horbelt, M., Antus, B., Wang, M., Exton, M.S. Influence of alternatively and classically activated macrophages on fibrogenic activities of human fibroblasts. Cell. Immunol. 2000; 204:19–28.  Kodelja, V., Muller, C., Tenorio, S., Schebesch, C., Orfanos, C.E., Goerdt, S. Differences in angiogenic potential of classically vs. alternatively activated macrophages. Immunobiology. 1997; 197:478–493.  El Kasmi KC, Qualls JE, Pesce JT, Smith AM, Thompson RW, Henao-Tamayo M, Basaraba RJ, König T, Schleicher U, Koo MS, Kaplan G, Fitzgerald KA, Tuomanen EI, Orme IM, Kanneganti TD, Bogdan C, Wynn TA, Murray PJ. Toll-like receptor- induced arginase 1 in macrophages thwarts effective immunity against intracellular pathogens. Nat Immunol. 2008; 9:1399-406.  Pesce JT, Ramalingam TR, Mentink-Kane MM, Wilson MS, El Kasmi KC, Smith AM, Thompson RW, Cheever AW, Murray PJ, Wynn TA Arginase-1-expressing macrophages suppress Th2 cytokine-driven inflammation and fibrosis. PLoS Pathog. 2009; 5(4):e1000371.  Herbert DR, Orekov T, Roloson A, Ilies M, Perkins C, O'Brien W, Cederbaum S, Christianson DW, Zimmermann N, Rothenberg ME, Finkelman FD. Arginase I suppresses IL-12/IL-23p40-driven intestinal inflammation during acute schistosomiasis. J Immunol. 2010; 184:6438-46.  Varin A, Mukhopadhyay S, Herbein G, Gordon S. Alternative activation of macrophages by IL-4 impairs phagocytosis of pathogens but potentiates microbial-induced signalling and cytokine secretion. Blood. 2010; 115:353-62.  Varin A, Gordon S. Alternative activation of macrophages: immune function and cellular biology. Immunobiology. 2009; 214:630-41.  Nathan C, Ding A. SnapShot: Reactive Oxygen Intermediates (ROI). Cell. 2010; 140:951-951.  Cassatella MA, Bazzoni F, Flynn RM, Dusi S, Trinchieri G, Rossi F. Molecular basis of interferon-gamma and lipopolysaccharide enhancement of phagocyte respiratory burst capability. Studies on the gene expression of several NADPH oxidase components. J Biol Chem. 1990; 265:20241-6.  Dewas C, Dang PM, Gougerot-Pocidalo MA, El-Benna J. TNF-alpha induces phosphorylation of p47(phox) in human neutrophils: partial phosphorylation of p47phox is a common event of priming of human neutrophils by TNF-alpha and granulocyte-macrophage colony-stimulating factor. J Immunol. 2003; 171:4392-8.  Decoursey TE, Ligeti E. Regulation and termination of NADPH oxidase activity. Cell Mol Life Sci. 2005; 62:2173-93.  El-Benna J, Dang PM, Gougerot-Pocidalo MA. Priming of the neutrophil NADPH oxidase activation: role of p47phox phosphorylation and NOX2 mobilization to the plasma membrane. Semin Immunopathol. 2008; 30:279-89.  Dang PM, Stensballe A, Boussetta T, Raad H, Dewas C, Kroviarski Y, Hayem G, Jensen ON, Gougerot-Pocidalo MA, El- Benna J. A specific p47phox –serine phosphorylated by convergent MAPKs mediates neutrophil NADPH oxidase priming at inflammatory sites. J Clin Invest. 2006; 116:2033-43.  Dang PM, Elbim C, Marie JC, Chiandotto M, Gougerot-Pocidalo MA, El-Benna J. Anti-inflammatory effect of interleukin-10 on human neutrophil respiratory burst involves inhibition of GM-CSF-induced p47PHOX phosphorylation through a decrease in ERK1/2 activity. FASEB J. 2006; 20:1504-6.  Bussolati B, Mariano F, Montrucchio G, Piccoli G, Camussi G. Modulatory effect of interleukin-10 on the production of platelet-activating factor and superoxide anions by human leukocytes. Immunology. 1997; 90:440-7.  Capsoni F, Minonzio F, Ongari AM, Carbonelli V, Galli A, Zanussi C. Interleukin-10 down-regulates oxidative metabolism and antibody-dependent cellular cytotoxicity of human neutrophils. Scand J Immunol. 1997; 45:269-75.  Kemlev SG, Palmer C. Interleukin-10 inhibits endotoxin-induced pro-inflammatory cytokines in microglial cell cultures. J Neuroimmunol. 2005; 162:71-80.  Qian L, Block ML, Wei SJ, Lin CF, Reece J, Pang H, Wilson B, Hong JS, Flood PM. Interleukin-10 protects lipopolysaccharide-induced neurotoxicity in primary midbrain cultures by inhibiting the function of NADPH oxidase. J Pharmacol Exp Ther. 2006; 319:44-52.  Qian L, Wei SJ, Zhang D, Hu X, Xu Z, Wilson B, El-Benna J, Hong JS, Flood PM. Potent anti-inflammatory and neuroprotective effects of TGF-beta1 are mediated through the inhibition of ERK and p47phox-Ser345 phosphorylation and translocation in microglia. J Immunol. 2008; 181:660-8.  Tyler KM, Engman DM. The life cycle of Trypanosoma cruzi revisited. Int J Parasitol. 2001; 31:472-81.  WHO 2010, Chagas disease (American trypanosomiasis) Fact sheet N°340, June 2010. http://www.who.int/mediacentre/factsheets/fs340/en/index.html.  Gironès N, Cuervo H, Fresno M. Trypanosoma cruzi-induced molecular mimicry and Chagas' disease. Curr Top Microbiol Immunol. 2005; 296:89-123  Savino W, Villa-Verde DM, Mendes-da-Cruz DA, Silva-Monteiro E, Perez AR, Aoki Mdel P, Bottasso O, Guiñazú N, Silva- Barbosa SD, Gea S. Cytokines and cell adhesion receptors in the regulation of immunity to Trypanosoma cruzi. Cytokine Growth Factor Rev. 2007; 18:107-24.  Tarleton RL. Immune system recognition of Trypanosoma cruzi. Curr Opin Immunol. 2007; 19:430-4.  Alvarez MN, Piacenza L, Irigoín F, Peluffo G, Radi R. Macrophage-derived peroxynitrite diffusion and toxicity to Trypanosoma cruzi. Arch Biochem Biophys. 2004; 432:222-32.  Alvarez MN, Peluffo G, Piacenza L, Radi R. Intraphagosomal peroxynitrite as a macrophage-derived cytotoxin against internalized Trypanosoma cruzi: consequences for oxidative killing and role of microbial peroxiredoxins in infectivity. J Biol Chem. 2011; 286:6627-40.  Cardoni RL, Antunez MI, Morales C, Nantes IR. Release of reactive oxygen species by phagocytic cells in response to live parasites in mice infected with Trypanosoma cruzi. Am J Trop Med Hyg. 1997; 56:329-34.  Carrera-Silva EA, Guiñazu N, Pellegrini A, Cano RC, Arocena A, Aoki MP, Gea S. Importance of TLR2 on hepatic immune and non-immune cells to attenuate the strong inflammatory liver response during Trypanosoma cruzi acute infection. PLoS Negl Trop Dis. 2010; 4(11):e863.  Melo R.C., Fabrino D.L., D’Avila H., Teixeira H.C. Ferreira A. P. “Production of hydrogen peroxide by peripheral blood monocytes and speciﬁc macrophages during experimental infection with Trypanosoma cruzi in vivo.” Cell Biology International. 2003; 27:853–861.  Celentano AM, González Cappa SM. Induction of macrophage activation and opsonizing antibodies by Trypanosoma cruzi subpopulations. Parasite Immunol. 1992; 14:155-67.  Hideko Tatakihara VL, Cecchini R, Borges CL, Malvezi AD, Graça-de Souza VK, Yamada-Ogatta SF, Rizzo LV, Pinge-Filho P. Effects of cyclooxygenase inhibitors on parasite burden, anemia and oxidative stress in murine Trypanosoma cruzi infection. FEMS Immunol Med Microbiol. 2008; 52:47-58.  Wen JJ, Vyatkina G, Garg N. Oxidative damage during chagasic cardiomyopathy development: role of mitochondrial oxidant release and inefficient antioxidant defense. Free Radic Biol Med. 2004; 37:1821-33.  Wen JJ, Garg NJ. Mitochondrial generation of reactive oxygen species is enhanced at the Q(o) site of the complex III in the myocardium of Trypanosoma cruzi-infected mice: beneficial effects of an antioxidant. J Bioenerg Biomembr. 2008; 40:587-98.  Guiñazú N, Carrera-Silva EA, Becerra MC, Pellegrini A, Albesa I, Gea S. Induction of NADPH oxidase activity and reactive oxygen species production by a single Trypanosoma cruzi antigen. Int J Parasitol. 2010; 40:1531-8.  Vespa GN, Cunha FQ, Silva JS. Nitric oxide is involved in control of Trypanosoma cruzi-induced parasitemia and directly kills the parasite in vitro. Infect Immun. 1994; 62:5177-82.  Francisco AF, de Abreu Vieira PM, Arantes JM, Silva M, Pedrosa ML, Elói-Santos SM, Martins-Filho OA, Teixeira-Carvalho A, Araújo MS, Tafuri WL, Carneiro CM. Increase of reactive oxygen species by desferrioxamine during experimental Chagas' disease. Redox Rep. 2010; 15:185-90  Petray P, Rottenberg ME, Grinstein S, Orn A. Release of nitric oxide during the experimental infection with Trypanosoma cruzi. Parasite Immunol. 1994; 16:193-9.  Malvezi AD, Cecchini R, de Souza F, Tadokoro CE, Rizzo LV, Pinge-Filho P. Involvement of nitric oxide (NO) and TNF- alpha in the oxidative stress associated with anemia in experimental Trypanosoma cruzi infection. FEMS Immunol Med Microbiol. 2004; 41:69-77.  Cummings KL, Tarleton RL. Inducible nitric oxide synthase is not essential for control of Trypanosoma cruzi infection in mice. Infect Immun. 2004; 72:4081-9.  Camargo MM, Andrade AC, Almeida IC, Travassos LR, Gazzinelli RT.Glycoconjugates isolated from Trypanosoma cruzi but not from Leishmania species membranes trigger nitric oxide synthesis as well as microbicidal activity in IFN-gamma-primed macrophages. J Immunol. 1997 Dec 15;159(12):6131-9.  Fernandez-Gomez R, Esteban S, Gomez-Corvera R, Zoulika K, Ouaissi A. Trypanosoma cruzi: Tc52 released protein-induced increased expression of nitric oxide synthase and nitric oxide production by macrophages. J Immunol. 1998; 160:3471-9.  Quanquin NM, Galaviz C, Fouts DL, Wrightsman RA, Manning JE. Immunization of mice with a TolA-like surface protein of Trypanosoma cruzi generates CD4(+)T-cell-dependent parasiticidal activity. Infect Immun. 1999; 67:4603-12  Ramos-Ligonio A, López-Monteon A, Talamás-Rohana P, Rosales-Encina JL. Recombinant SSP4 protein from Trypanosoma cruzi amastigotes regulates nitric oxide production by macrophages. Parasite Immunol. 2004; 26:409-18.  Guiñazú N, Pellegrini A, Carrera-Silva EA, Aoki MP, Cabanillas AM, Gìronés N, Fresno M, Cano R, Gea S. Immunisation with a major Trypanosoma cruzi antigen promotes pro-inflammatory cytokines, nitric oxide production and increases TLR2 expression. Int J Parasitol. 2007; 37:1243-54.  Araújo AF, de Alencar BC, Vasconcelos JR, Hiyane MI, Marinho CR, Penido ML, Boscardin SB, Hoft DF, Gazzinelli RT, Rodrigues MM. CD8+-T-cell-dependent control of Trypanosoma cruzi infection in a highly susceptible mouse strain after immunization with recombinant proteins based on amastigote surface protein 2. Infect Immun. 2005; 73:6017-25.  Ouaissi A, Guilvard E, Delneste Y, Caron G, Magistrelli G, Herbault N, Thieblemont N, Jeannin P. The Trypanosoma cruzi Tc52-released protein induces human dendritic cell maturation, signals via Toll-like receptor 2, and confers protection against lethal infection. J Immunol. 2002; 168:6366-74.  Campos MA, Almeida IC, Takeuchi O, Akira S, Valente EP, Procópio DO, Travassos LR, Smith JA, Golenbock DT, Gazzinelli RT. Activation of Toll-like receptor-2 by glycosylphosphatidylinositol anchors from a protozoan parasite. J Immunol. 2001  Campos MA, Closel M, Valente EP, Cardoso JE, Akira S, Alvarez-Leite JI, Ropert C, Gazzinelli RT. Impaired production of proinflammatory cytokines and host resistance to acute infection with Trypanosoma cruzi in mice lacking functional myeloid differentiation factor 88. J Immunol. 2004; 172:1711-8.  Giordanengo L, Guiñazú N, Stempin C, Fretes R, Cerbán F, Gea S. Cruzipain, a major Trypanosoma cruzi antigen, conditions the host immune response in favor of parasite. Eur J Immunol. 2002; 32:1003-11.  Stempin C, Giordanengo L, Gea S, Cerbán F. Alternative activation and increase of Trypanosoma cruzi survival in murine macrophages stimulated by cruzipain, a parasite antigen. J Leukoc Biol. 2002; 72:727-34.  Stempin CC, Tanos TB, Coso OA, Cerbán FM. Arginase induction promotes Trypanosoma cruzi intracellular replication in Cruzipain-treated J774 cells through the activation of multiple signaling pathways. Eur J Immunol. 2004; 34:200-9.  Chen L, Watanabe T, Watanabe H, Sendo F. Neutrophil depletion exacerbates experimental Chagas’ disease in BALB/c, but protects C57BL/6 mice through modulating the Th1/Th2 dichotomy in differ- ent directions. Eur J Immunol 2001;31:265–75.  Denkers EY, Butcher BA, Del Rio L, Bennouna S, Neutrophils. dendritic cells and Toxoplasma. Int J Parasitol 2004;34:411–21.  Tacchini-Cottier F, Zweifel C, Belkaid Y, Mukankundiye C, Vasei M, Launois P, et al. An immunomodulatory function for neutrophils during the induction of a CD4+Th2 response in BALB/c mice infected with Leishmania major. J Immuno. 2000; 165:2628–36.  Fadok VA, Bratton DL, Konowal A, Freed PW, Westcott JY, Henson PM. Macrophages that have ingested apoptotic cells in vitro inhibit proinflammatory cytokine production through autocrine/paracrine mechanisms involving TGF-beta, PGE2, and PAF. J Clin Invest. 1998; 101:890–8.  Freire-de-Lima CG, Nascimento DO, Soares MB, Bozza PT, Castro- Faria-Neto HC, de Mello FG, et al. Uptake of apoptotic cells drives the growth of a pathogenic trypanosome in macrophages. Nature. 2000; 403:199–203.  DosReis GA, Ribeiro-Gomes FL, Guillermo LV, Lopes MF. Cross-talk between apoptosis and cytokines in the regulation of parasitic infection. Cytokine Growth Factor Rev. 2007; 18:97-105.  Carrera L, Gazzinelli RT, Badolato R, Hieny S, Muller W, Kuhn R, Sacks DS. Leishmania promastigotes selectively inhibit interleukin 12 induction in bone marrow-derived macrophages from susceptible and resistant mice. J. Exp. Med. 1996; 183:515–526.  Denkers EY, Kim L, Butcher BA. In the belly of the beast: subversion of macrophage proinflammatory signalling cascades during Toxoplasma gondii infection. Cell Microbiol. 2003; 5:75–83.  Butcher BA, Kim L, Johnson PF, Denkers EY. Toxoplasma gondii tachyzoites inhibit proinflammatory cytokine induction in infected macrophages by preventing nuclear translocation of the transcription factor NF-kappa B. J. Immunol. 2001; 167:2193–2201.  Planelles L, Thomas M, Maranon C, Morell M, Lopez M. Differential CD86 and CD40 co-stimulatory molecules and cytokine expression pattern induced by Trypanosoma cruzi in APCs from resistant or susceptible mice. Clin Exp Immunol; 2003;131:41–7.  Guiñazú N, Pellegrini A, Giordanengo L, Aoki M, Rivarola H, Cano R, et al. Immune response to a major Trypanosoma cruzi antigen, cruzipain, is differentially modulated in C57BL/6 and BALB/c mice. Microbes Infect. 2004;6:1250–8.  Carrera-Silva EA, Cano RC, Guinazu N, Aoki MP, Pellegrini A, et al. (2008) TLR2, TLR4 and TLR9 are differentially modulated in liver lethally injured from BALB/c and C57BL/6 mice during Trypanosoma cruzi acute infection. Mol Immunol. 45:3580–3588.  Brodskyn C, Patricio J, Oliveira R, Lobo L, Arnholdt A, Mendonça-Previato L, Barral A, Barral-Netto M. Glycoinositolphospholipids from Trypanosoma cruzi interfere with macrophages and dendritic cell responses. Infect Immun. 2002; 70:3736-43.  Padilla AM, Simpson LJ, Tarleton RL. Insufficient TLR activation contributes to the slow development of CD8+ T cell responses in Trypanosoma cruzi infection. J Immunol. 2009; 183:1245–1252.
Pages to are hidden for
"129"Please download to view full document


UK Chancellor Jeremy Hunt will make major tax and spend decisions today as calls grow for help with household finances, an expanded levy on oil and gas, and a decision on major job-creating projects across the north and north-east.
The Autumn Statement presents an opportunity for the new chancellor to draw a line under the chaos of Liz Truss’ premiership.
Against a backdrop of financial uncertainty, here are some of the problems the Tories need to tackle.
1. Cost of living
Mr Hunt could choose to tie benefits and the state pension to inflation, as previously suggested.
The prime minister also pledged in May to raise benefits, when he was still serving as chancellor, but he has remained tight-lipped since then.
Further rises in gas and electricity bills sent UK inflation to its highest level for 41 years, according to official figures released on Wednesday.
The chancellor is expected to boost the national living wage by almost 10% from £9.50 to £10.40 an hour, helping more than 2.5 million Brits.
He is also said to be planning to announce another round of cost-of-living payments of up to £1,100 for the most vulnerable households.
But Mr Hunt has vowed to deliver a package of tax hikes and spending cuts in order to face the economic “storm”.
2. Long-awaited freeports decision
The UK Government has refused for weeks to reveal its long-awaited decision on which two Scottish bids will secure special tax status as “freeports”.
There is speculation the budget could pave the way for a decision being announced.
The announcement was expected in the summer – and we previously revealed a decision had even been taken behind closed doors more than a month ago.
It was also understood plans from former Prime Minister Liz Truss to give low-tax investment status to the losing bids had caused a hold-up.
Aberdeen South MP Stephen Flynn says the silence from the UK Government “has been deafening”.
He wrote to the chancellor this week urging him to commit to the North East Scotland bid as part of his Autumn Statement.
Four other bids were lodged by groups in Inverness and Cromarty, Orkney, Clyde and Forth.
3. Windfall tax
Oil and gas firms are bracing themselves for a surge in the windfall tax from 25% to 35% and an extension of the scheme from 2025 to 2028.
Mr Hunt is also anticipated to widen the Energy Price Levy to include electricity generators.
Demands for a deeper levy were reignited last month when Shell reported global profits of £8.2 billion – its second-highest quarterly profit on record.
There is a growing clamour for fossil fuel producers to stump up cash at a time when families are struggling to make ends meet.
However, oil and gas companies have issued public pleas to the chancellor urging him to rethink the much-expected changes.
They warn that taxing oil and gas firms in the UK for the second time in six months would risk “driving investment out of the UK altogether”.


text:
The Latta Town Council has voted to make its small South Carolina community "council strong/mayor weak" in response to Mayor Earl Bullard's decision to fire Police Chief Crystal Moore -- a 20-year veteran of the force who was also the town's first openly gay police chief. Many say the move had nothing to do with Moore's job performance, but was driven by Bullard's homophobia.
Bullard fired Moore last week after giving her seven reprimands in a single day, which another council member called "questionable.” According to local news station WBTW, the reprimands included running “unauthorized” background checks, “questioning authority of supervisor,” “questioning authority of mayor,” “failure to maintain order” and “contacting the news media.”
Moore, joined by members of the council and many in her community, believes that Bullard fired her because she is gay. Bullard was recently recorded by a fellow council member while making anti-LGBTQ remarks.
“I would much rather have ... and I will say this to anybody’s face … somebody who drank and drank too much taking care of my child than I had somebody whose lifestyle is questionable around children," Bullard said during the recording. "I’m not going to let 2 women stand up there and hold hands and let my child be aware of it. And I’m not going to see them do it with 2 men neither. I’m not going to do it. Because that ain’t the way the world works.”
Moore, who has served her community for two decades, is shocked by the loss of her position, telling WBTW, "I can't believe that we still have no equal rights. That's the biggest issue. I've been harassed, intimidated. This is the first time it's been this public. I'd tried living a quiet decent life and do what I'm supposed to."
The referendum to make Latta "council strong/mayor weak" will next be voted on by city residents, effectively stripping Bullard of some of his power.
But the question of whether Moore will be reinstated remains, though some members of the town council say they are meeting to get Moore back on the force.