The potential of live attenuated vaccines against Cutaneous Leishmaniasis.

Cutaneous Leishmaniasis is a serious public health problem, typically affecting poor populations with limited access to health care. Control is largely dependent on chemotherapies that are inefficient, costly and challenging to deliver. Vaccination is an attractive and feasible alternative because long-term protection is typical in patients who recover from the disease. No human vaccine is yet approved for use, but several candidates are at various stages of testing. Live attenuated parasites, which stimulate long-term immune protection, have potential as effective vaccines, and their challenges relating to safety, formulation and delivery can be overcome. Here we review current data on the potential of live attenuated Leishmania vaccines and discuss possible routes to regulatory approval.

Enhanced sonocatalytic performance of ZnTi nano-layered double hydroxide by substitution of Cu (II) cations.

In this research, a series of CuZnTi-LDHs with different Cu2+/Zn2+ molar ratio were synthesized by co-precipitation method with the purpose of improving the sonocatalytic performance of ZnTi-LDH. All the LDH samples were synthesized by a facile co-precipitation process. The as-prepared LDHs were characterized by Powder X-ray diffraction (XRD), Field emission-scanning electron microscopy (FESEM), Transition electron microscopy (TEM), Brunauer-Emmelt-Teller (BET) analysis, and UV-visible diffuse reflectance spectroscopy (DRS) analysis. The results showed that Cu2+ substitution can significantly enhance the sonocatalytic properties of ZnTi-LDH. The Methylene blue degradation percentage over ZnTi-LDH reached 30% in 90 min, whilst this percentage reaches 71% over CuZnTi-LDH (1:1). The role of the Cu2+ incorporation on the observed enhancement in sonocatalytic performance was revealed by investigating the effect of radical scavengers on degradation efficiency and DRS spectra of ZnTi-LDH and CuZnTi-LDH (1:1). Benzoquinone (BQ), ammonium oxalate and tert-Bu lead to 22.5%, 53.5% and 74.6% decrease in degradation percentage by CuZnTi-LDH (1:1). However, the degradation efficiency showed 16.6%, 3.3% and 63.3% reduction in the presence of BQ, ammonium oxalate and tert-Bu respectively, in dye degradation by ZnTi-LDH. DRS spectra demonstrated that the band gap of the LDH decreases by Cu2+ substitution. The effect of operational parameters on sonodegradation was investigated as well. The kinetics of sonodegradation reaction obeyed the first order reaction kinetics with R2 of 0.95.

TLR3 and its roles in the pathogenesis of type 2 diabetes.

Type 2 diabetes (T2D) is the most prevalent non—infectious disease and leads to several complications including nephropathy and retinopathy. The mechanisms and signaling molecules responsible for the development and progression of T2D, as well as its associated complications are yet to be identified. It would appear that genetic backgrounds and immunological parameters of people susceptible to T2D may play important roles in induction of T2D. TLRs participate in several cellular pathways which can induce activation of proliferation. However, in contradiction, these pathways can also be associated with apoptosis. The multiple roles of TLRs and their signaling molecules associated with T2D pathways makes them candidates for the induction of immune—regulated diseases like T2D. TLR3 has been identified as an intracellular ligand and subsequently activates signaling molecules via the TRIF pathway. Therefore, the alteration of expression of TLR3 and their functions may lead to inappropriate induction of immune system functions that are related to T2D disease. The aim of this review was to collect recent data regarding the roles of TLR3 in the progression and pathogenesis of T2D.

Circulating levels of interleukin-35 in patients with multiple sclerosis: evaluation of the influences of FOXP3 gene polymorphism and treatment program.

The regulatory T (Treg) cells play a major role in the control of the autoimmunity and inflammation, and IL-35 has been described as an immunosuppressive cytokine that is mainly produced by CD4(+)FOXP3(+) Treg cells. The aim of this study was to evaluate the serum levels of IL-35 and a single nucleotide polymorphism (SNP), rs3761548, in FOXP3 gene in patients with multiple sclerosis. The blood samples were collected from 140 multiple sclerosis (MS) patients (including 51 untreated and 89 treated patients) and 140 healthy subjects as a control group. The serum levels of IL-35 were measured by ELISA. The DNA was analyzed for SNP rs3761548 in FOXP3 gene using SSP-PCR. There was no significant difference between untreated MS patients and control group regarding the mean serum levels of IL-35, although this parameter was higher in untreated patients. However, the mean serum level of IL-35 in treated MS patients was significantly higher than that in the control group (P < 0.008). The mean serum levels of IL-35 in patients who were treated with interferon-ß, methylprednisolone, or with the both interferon-ß and methylprednisolone were significantly higher than that in the healthy group (P < 0.01, P < 0.01, and P < 0.2, respectively). The frequencies of AA and AC genotypes at rs3761548 in the FOXP3 gene were significantly higher in MS group as compared with healthy subjects (P < 0.05). The frequency of CC genotype at rs3761548 was significantly lower in the MS group in comparison with healthy control subjects (P < 0.001). Moreover, the frequency of A allele was significantly higher whereas the frequency of C allele was significantly lower in MS patients in comparison to healthy subjects (P < 0.001). The mean serum level of IL-35 was significantly lower in MS patients or healthy subjects with AA genotype as compared with those with CC genotype at rs3761548 in FOXP3 gene (P < 0.01 and P < 0.001, respectively). These results showed higher serum levels of IL-35 in treated MS patients representing that the benefit effects of treatment may in part performed through the upregulation of the IL-35 production. The SNP rs3761548 may influence the susceptibility to MS disease and the serum levels of IL-35.

Dogs vaccinated with gentamicin-attenuated Leishmania infantum or infected with wild-type parasite can be distinguished by Western blotting.

An attenuated line of Leishmania infantum (the H-line), developed through exposure to gentamicin, has been shown to protect dogs against canine visceral leishmaniasis. A specific diagnostic test to differentiate dogs vaccinated with the attenuated line from dogs infected with L. infantum wild-type (L. infantum WT) could be a valuable tool in evaluating the effectiveness of canine vaccination. In this study, 28 healthy dogs were allocated into four groups. In Group I and Group II (eight dogs per group), dogs were immunized subcutaneously (s.c.) with L. infantum H-line, and the dogs of Group II challenged s.c. with L. infantum WT, at 2 months post-immunization. In Group III, eight animals were challenged s.c. with L. infantum WT, and four dogs of Group IV were injected s.c. with PBS. We found that sera from vaccinated dogs recognize a 21 kDa antigen of promastigotes of L. infantum H-line but not of L. infantum WT, whereas sera from unvaccinated dogs challenged with L. infantum WT, recognized a 21 kDa antigen of promastigotes of L. infantum WT but not of L. infantum H-line. Sera from dogs challenged with L. infantum WT with prior vaccination with L. infantum H-line, recognized a 21 kDa antigen of both L. infantum WT and L. infantum H-line. These results suggest that the Western blot analysis of antibodies against 21 kDa antigens of L. infantum H-line and WT may be a useful technique for distinguishing between dogs vaccinated with L. infantum H-line and dogs naturally infected with L. infantum WT.

Higher circulating levels of chemokine CCL20 in patients with multiple sclerosis: evaluation of the influences of chemokine gene polymorphism, gender, treatment and disease pattern.

Chemokines play an important role in the autoimmune diseases. The aim of this study was to investigate the levels of CCL20 and a polymorphism [-786C > T (rs6749704)] in the chemokine gene in patients with multiple sclerosis (MS). The blood samples were collected from 135 MS patients and 135 healthy subjects as a control group. The patients have relapsing-remitting (RRMS; n = 65), primary progressive (PPMS; n = 47), secondary progressive (SPMS; n = 35) or progressive relapsing (PRMS; n = 14) patterns. The serum levels of CCL20 were measured by ELISA. The DNA was analyzed for CCL20 polymorphism using PCR-RLFP. The mean serum levels of CCL20 in the MS group were significantly higher than in the healthy group (P < 0.001). In patients with a SPMS pattern, the frequency of CT genotype at rs6749704 (24.3 %) was significantly lower as compared to patients with other patterns (42.8 %; P < 0.04). No significant differences were observed between subjects with different genotypes in rs6749704 regarding the CCL20 levels. The mean serum levels of CCL20 in both newly diagnosed and previously diagnosed patients was significantly higher than in the healthy group (P < 0.05 and 0.001, respectively). The mean serum levels of CCL20 in patients with RRMS, SPMS and PPMS patterns were significantly higher than in the healthy group (P < 0.004, P < 0.04, and 0.05, respectively). The levels of CCL20 in untreated patients and in patients who received interferon-ß, methylprednisolone or the combination of interferon-ß plus methylprednisolone were higher as compared to the control group (P < 0.05, P < 0.03, P < 0.005, and P < 0.05, respectively). These results showed higher levels of CCL20 in patients that represent that the chemokine may play an important role in the pathogenesis of MS. The rs6749704 polymorphism was an associated SPMS pattern. The levels of CCL20 were not influenced by gender, disease pattern and treatment.

Lower serum levels of Th2-related chemokine CCL22 in women patients with multiple sclerosis: a comparison between patients and healthy women.

Chemokines play a major role in autoimmune diseases such as multiple sclerosis (MS). Gender also affects the susceptibility and course of MS. The aim of this study was to investigate the serum levels of the macrophage-derived chemokine (CCL22) in women and men patients with MS. Blood samples were collected from 135 healthy subjects (35 men and 100 women) and 135 MS patients (29 men and 136 women; 47 newly diagnosed and 88 treated patients and have relapsing-remitting (RRMS; n = 65), secondary progressive (SPMS; n = 37), primary progressive (PPMS; n = 19), or progressive relapsing (PRMS; n = 14) patterns). The serum levels of CCL22 were measured by ELISA. The difference of the mean serum levels of CCL22 between the newly diagnosed MS men and healthy men was not significant, but in newly diagnosed MS women, the mean serum levels of CCL22 were significantly lower than those in treated MS women and healthy women (P < 0.006 and P < 0.0001, respectively). The differences of the mean CCL22 levels between men patients with different treatment programs were not significant, but the mean CCL22 levels were significantly higher in women treated with interferon-ß or the combination of interferon-ß plus methylprednisolone as compared to untreated women patients (P < 0.01 and P < 0.05, respectively). The CCL22 levels were also significantly higher in women with RRMS and PRMS patterns in comparison to healthy women (P < 0.05 and P < 0.01, respectively). These results showed lower levels of CCL22 in women patients which represents that the reduction in CCL22 levels may play an important role in the pathogenesis of the disease in women. In women patients, the levels of CCL22 were influenced by disease pattern and treatment.

Association of PTPN22 rs2476601 and EGFR rs17337023 Gene polymorphisms and rheumatoid arthritis in Zahedan, Southeast Iran.

In this study we aimed to evaluate the possible association of PTPN22 rs2476601 as well as epidermal growth factor receptor (EGFR) rs17337023 gene polymorphism and rheumatoid arthritis (RA) in a sample of Iranian population. This case-control study was performed on 120 patients with RA and 120 healthy subjects. Genomic DNA was extracted from whole blood and PTPN22 rs2476601 and EGFR rs17337023 polymorphisms were determined using tetra amplification refractory mutation system-polymerase chain reaction (T-ARMS-PCR). The results showed that PTPN22 rs2476601 CT genotype as well as rs2476601 T allele was a risk factor for susceptibility to RA (OR=5.89 95%CI = 1.78-19.48, P = 0.004 and OR = 4.78, 95%CI = 1.59-14.35, P = 0.003, respectively). We also found that EGFR rs17337023 AT and rs17337023 TT genotypes were risk factor for susceptibility to RA (OR = 9.94 95%CI = 3.65-26.73, P < 0.001 and OR = 3.66, 95%CI = 1.46-9.15, P = 0.005, respectively). In addition the EGFR rs17337023 T allele was a risk for predisposition to RA (OR = 1.56, 95%CI=1.06-2.30, P = 0.030). In conclusion, we found an association between PTPN22 rs2476601 and EGFR rs17337023 polymorphisms and the risk of RA in a sample of Iranian population.

Gentamicin-attenuated Leishmania infantum: cellular immunity production and protection of dogs against experimental canine leishmaniasis.

An attenuated line of Leishmania infantum (L. infantum H-line) has been established by culturing promastigotes in vitro under gentamicin pressure. Here, we show that L. infantum H-line induced significantly higher levels of IFN-γ and lower levels of IL-10 compared with those in dogs infected with L. infantum wild type (WT). Anti-Leishmania-specific total IgG, IgG1, and IgG2 antibodies were present in the serum of all infected dogs, with levels of IgG2 subclass highest in the sera of dogs inoculated with L. infantum H-line. Relatively high levels of IgG1 were found in the sera of dogs infected with L. infantum WT. Six of seven dogs immunized intradermally (i.d.) with the attenuated line later showed a positive skin test to leishmanin, whereas the dogs infected with L. infantum WT did not. No clinical abnormalities were observed, and no parasites found in the visceral organs of the dogs inoculated intravenously (i.v.) with L. infantum H-line over 24 months post-inoculation. Dogs which had been immunized with L. infantum H-line i.d. 12 months previously were protected against challenge with L. infantum WT. These data suggest that the L. infantum H-line was safe and induced a protection which is correlated with cellular immunity in dogs.

Collagen characteristics and organization during the progression of cholesterol-induced atherosclerosis in Japanese quail.

This study reports the concentration of collagen and its hydroxypyridinoline crosslinks, collagen fibril organization in the dorsal aortas, and systolic blood pressure during the progression of atherosclerosis in Japanese quail selected for cholesterol-induced atherosclerosis. The quail were placed on either a control or 0.5% cholesterol-added diet at approximately 16 weeks of age. The concentration of total collagen did not change in the control arteries during the course of the study, whereas at 5 and 10 weeks of cholesterol feeding, collagen levels decreased in the cholesterol-fed birds. Hydroxypyridinoline concentration increased during the duration of the study in the cholesterol-fed birds and by 15 and 20 weeks of cholesterol feeding, levels were significantly increased over those observed in the control arteries. Transmission electron microscopy showed changes in the organization of collagen fibrils. Increased systolic blood pressure was noted beginning at 10 weeks of cholesterol feeding, which is suggestive of other systemic changes induced by hypercholesterolemia. These results demonstrated remodeling of the collagen component of the dorsal aorta extracellular matrix during the progression of atherosclerosis and are suggestive of other systemic cardiovascular system changes.

Testosterone increases blood pressure and cardiovascular and renal pathology in spontaneously hypertensive rats.

The objective of this paper was to test the hypothesis that testosterone (T) raises blood pressure (BP), which is associated with increased coronary adventitial collagen, whereas the hemodynamic force of BP increases the coronary media:lumen ratio. Five treatment groups of spontaneously hypertensive rat (SHR) were established (n = 8-10 per group): controls; hydralazine (HYZ); castration; castration + HYZ; and castration + HYZ + T + captopril. At 12 weeks of age, the castrate + HYZ group was divided so that the mean BP was the same in both groups (162 mmHg). Both groups continued to receive HYZ treatment; however one group received T implants. Also, at 12 weeks of age the castrate + HYZ + T + captopril group received T implants. BP in the HYZ group was reduced compared with controls (192 mmHg vs 218 mmHg, p < 0.01). Castration lowered BP to 170 mmHg (p < 0.01) compared with controls. However, T implants increased BP by 15 mmHg (p < 0.02) in the castrate + HYZ group and by 44 mmHg in the castrate + HYZ + captopril group (p < 0.01). Captopril in combination with HYZ significantly reduced BP compared with controls but T replacement increased BP and coronary collagen deposition in spite of HYZ and captopril treatment.

Nd:YAG laser corneal disruption as adjuvant treatment for infectious crystalline keratopathy.

PURPOSE: To report on the use of the Nd:YAG laser corneal disruption in the treatment of infectious crystalline keratopathy. METHOD: Case report. A 52-year-old woman with infectious crystalline keratopathy unresponsive to topical antibiotics was treated with an Nd:YAG laser to the intrastromal crystals. RESULTS: After Nd:YAG laser treatment, the infiltrate completely cleared within 4 weeks. CONCLUSIONS: Nd:YAG laser treatment may be effective in disrupting the protective glycocalyx matrix within the intrastromal crystals, rendering the bacteria susceptible to topical antibiotics. This treatment should be considered for patients with infectious crystalline keratopathy clinically resistant to topical antibiotics.

Symptomatic and asymptomatic visual loss in patients taking vigabatrin.

PURPOSE: To investigate the clinical, perimetric, and electrophysiologic findings in patients with visual field loss on long-term treatment with the antiepileptic medication vigabatrin. DESIGN: Consecutive observational case series. PARTICIPANTS: Forty-one consecutive subjects taking vigabatrin referred for screening ophthalmologic assessment were studied. Twelve subjects with evidence of peripheral visual field constriction are presented. METHODS: Twelve subjects with evidence of peripheral visual field constriction on 60-4 perimetry underwent central 30-2 and blue-on-yellow (B/Y) perimetry, as well as electroretinography (ERG), electro-oculography (EOG), and visual-evoked potential (VEP) testing. MAIN OUTCOME MEASURES: Visual acuity; fundus abnormalities; visual field loss; and ERG, EOG, or VEP abnormalities were the main outcome measures. RESULTS: Eight of the 12 subjects with constricted visual fields were asymptomatic. The central 30-2 perimetry demonstrated bilateral visual field constriction in 9 of 12 patients and the B/Y perimetry in 8 of 9 patients tested. Of the ten patients tested electrophysiologically, four had abnormal ERGs, five had abnormal EOGs, and three had delayed VEPs. CONCLUSIONS: The incidence of visual field constriction in patients taking vigabatrin may be higher, and asymptomatic visual field loss more common, than reported previously. The authors postulate a possible Muller cell dysfunction in the peripheral retina. Patients taking vigabatrin should have regular peripheral visual field examinations.

Neonatal sympathectomy reduces adult blood pressure and cardiovascular pathology in Y chromosome consomic rats.

The hypothesis was tested that the sympathetic nervous system (SNS) developmentally influences circulating testosterone (T), systolic blood pressure (SBP) and cardio-renal pathology in SHR/y animals. A sympathoplegic drug, guanethidine, and an antibody to nerve growth factor were administered to WKY and borderline hypertensive SHR/y male rats (n = 20/group) for the first 3 weeks of life; control groups (n = 20/group) received saline. SBP, serum T and luteinizing hormone (LH) were measured. SBP in the WKY and SHR/y sympathectomy (sympx) groups decreased 10mmHg (p < 0.001) and 50mmHg (p < 0.001), respectively, when compared to their control groups. Serum T levels in the sympx WKY group were lower (p < 0.01) than those in controls, and the rise of T typically observed in SHR/y from weeks 6-8 was delayed in the sympx SHR/y group, similar to the pattern in WKY. Serum LH levels were increased in the sympx WKY group, but not in the SHR/y group. Sympx caused a greater reduction in renal glomerular changes (p < 0.01), coronary artery collagen deposition (p < 0.01) and myocardial fibrosis (p < 0.01) in SHR/y than WKY rats. In conclusion, the SHR Y chromosome has a locus that enhances SNS activity, which can raise SBP and result in renal and cardiovascular tissue damage.

Management of submacular hemorrhage with intravitreal sulfur hexafluoride: a pilot study.

BACKGROUND: Some success has been reported with the intravitreal use of tissue plasminogen activator (tPA) and perfluoropropane gas in the management of large submacular hemorrhages. However, the dosage of tPA that has been used (100 micrograms) has a narrow margin of safety, and it remains to be shown that intravitreal tPA can cross the retina and effect subretinal clot lysis. We carried out a pilot study to evaluate the efficacy of intravitreally administered sulfur hexafluoride (SF6) gas alone in the management of large submacular hemorrhages secondary to age-related macular degeneration (AMD). METHODS: Three patients with large submacular hemorrhages secondary to AMD seen at a university-affiliated teaching hospital in Ottawa were treated with an intravitreal injection of 0.6 mL of SF6 gas. They were instructed to assume a prone position for 7 to 10 days. The patients were followed 3, 7, 14 and 28 days after the procedure and monthly thereafter for at least 6 months. Colour photography and fluorescein and indocyanine green angiography were performed immediately before and 2 weeks after the procedure and, thereafter, at the discretion of the treating ophthalmologist. RESULTS: In all three cases significant inferior displacement of the submacular blood was observed. Two patients showed an improvement of vision from counting fingers to 20/70 and to 20/200. In one case the submacular blood was displaced such that laser photocoagulation of a juxtafoveal choroidal neovascular membrane became possible. INTERPRETATION: The results suggest that intravitreally administered SF6 alone may have a role in the management of selected cases of neovascular AMD complicated by significant submacular hemorrhage. These results call into question the utility of adjunctive intravitreal tPA in such cases.

Spontaneously hypertensive rat Y chromosome increases indexes of sympathetic nervous system activity.

Previous studies from our laboratory have demonstrated that the Y chromosome from the spontaneously hypertensive rat (SHR) is responsible for a significant portion of the elevated blood pressure and also produces an earlier pubertal rise in plasma testosterone. We performed the following studies to determine whether the SHR Y chromosome raises blood pressure by sympathetic nervous system responses as measured by adrenal chromogranin A and plasma and tissue catecholamines. Male SHR from the University of Akron colony were studied from 5 to 20 weeks of age. Blood pressure was measured by tail-cuff, tail artery cannulation, and aortic telemetry (Data Sciences); acute (air stress) and chronic (territorial colony) social stressors were compared; blood was collected for determination of plasma catecholamines; and adrenal glands were analyzed at 15 weeks for catecholamines. Rats with the SHR Y chromosome had higher blood pressure and plasma norepinephrine than those with the normotensive Wistar-Kyoto (WKY) Y chromosome. However, the SHR Y chromosome did not significantly change responsiveness to acute or chronic stressors. Phentolamine and clonidine prevented the stress responses. Adrenal chromogranin A levels were elevated 37% and 40% and adrenal norepinephrine content 29% and 100% at 4 and 10 weeks of age, respectively, in rats with an SHR Y chromosome compared with WKY. Chemical sympathectomy normalized blood pressure in all strains and significantly reduced norepinephrine (36% to 41%) in all strains except in WKY, which already had a normal blood pressure. In conclusion, the SHR Y chromosome appears to increase the chronic sympathetic nervous system. A potential mechanism could be a Y locus that influences chronic sympathetic nervous system activity, which may reinforce neurohumoral factors and structural components of the vessel wall, accelerating the development of hypertension.

γ-Glutamyl semialdehyde and 2-amino-adipic semialdehyde: biomarkers of oxidative damage to proteins.

Reactive oxygen species are formed in the body by several natural processes and by induced oxidative stress. The reactive oxygen species may react with the various biomolecules of the body, including proteins. In order to assess the impact of oxidative damage to proteins, we have tried to identify oxidized amino acids in blood proteins which might serve as biomarkers of oxidative damage. When oxidative damage is induced into bovine serum albumin by metal-catalysed oxidation systems, the aldehyde groups formed can be derivatized by fluoresceinamine (FINH2). Following acid hydrolysis of FINH2-derivatized protein, two major oxidation products, γ-glutamyl semialdehyde (GGS) and 2-amino-adipic semialdehyde (AAS), were found and identified by HPLC and MS. Isolation and identification of oxidized amino acids from homopolymers (poly-Arg,-Pro,-Lys,-Trp or -Leu) confirmed that GGS can originate from Arg or Pro, while AAS is an oxidation product of Lys. When oxidative stress was induced in rats by treatments with t-butyl hydroperoxide or acrolein, rat plasma protein levels of GGS and AAS were found to be significantly higher compared with control rats. The AAS-content in serum albumin or in total plasma proteins collected from eight different mammalian species was found to be inversely proportional to their maximum lifespan potential. The content of AAS in plasma proteins of untreated adult rats showed a positive correlation with the age of the rat. In young rats a negative correlation with age was found for both GGS and AAS. We conclude that GGS or AAS may be useful novel biomarkers of oxidative damage to proteins in vivo.