The Route of Leishmania tropica Infection Determines Disease Outcome and Protection against Leishmania major in BALB/c Mice

Leishmania tropica is one of the causative agents of leishmaniasis in humans. Routes of infection have been reported to be an important variable for some species of Leishmania parasites. The role of this variable is not clear for L. tropica infection. The aim of this study was to explore the effects of route of L. tropica infection on the disease outcome and immunologic parameters in BALB/c mice. Two routes were used; subcutaneous in the footpad and intradermal in the ear. Mice were challenged by Leishmani major, after establishment of the L. tropica infection, to evaluate the level of protective immunity. Immune responses were assayed at week 1 and week 4 after challenge. The subcutaneous route in the footpad in comparison to the intradermal route in the ear induced significantly more protective immunity against L. major challenge, including higher delayed-type hypersensitivity responses, more rapid lesion resolution, lower parasite loads, and lower levels of IL-10. Our data showed that the route of infection in BALB/c model of L. tropica infection is an important variable and should be considered in developing an appropriate experimental model for L. tropica infections.     

Intra-erythrocyte Magnesium Is Associated with Gamma-Glutamyl Transferase in Obese Children and Adolescents

This study aims at determining the association between markers of hepatic injury and serum, urinary, and intra-erythrocyte magnesium concentrations and dietary magnesium intake in obese children and adolescents. In a case–control study, 42 obese children and adolescents (8–18 years) and 42 sex- and puberty-matched controls were studied. Serum, urinary, and intra-erythrocyte magnesium levels, indices of insulin sensitivity, and liver enzymes were measured. Dietary magnesium intake was assessed using a food frequency questionnaire. Obese children and adolescents exhibited insulin resistance as determined by a higher fasting insulin and the HOMA-IR (p < 0.001) and lower QUICKI indices (p = 0.001); in addition these subjects had significantly higher intra-erythrocyte magnesium (IEM) concentrations, than non-obese ones (3.99 ± 1.05 vs. 3.35 ± 1.26 mg/dL of packed cell; p = 0.015). Among liver enzymes, only gamma-glutamyl transferase (GGT) was significantly higher in obese than in non-obese subjects (22.7 ± 9.4 vs. 17.1 ± 7.9 U/l; p = 0.002). A positive association was found between GGT and IEM in both groups; however in multivariate analysis, in obese subjects, only GGT (p = 0.026) and, in non-obese subjects, only age (p = 0.006) remained as significant predictors of IEM. In conclusion, increased IEM concentration was seen in insulin-resistant obese children and adolescents; furthermore, serum GGT was associated with IEM, independently of body mass index and HOMA-IR.     

Cryopreservation of immature porcine testis tissue to maintain its developmental potential after xenografting into recipient mice

The purpose of this study was to develop effective strategies for cooling and cryopreservation of immature porcine testis tissue that maintain its developmental potential. Testes from 1-wk-old piglets (Sus domestica) were subjected to 1 of 12 cooling/cryopreservation protocols: as intact testes, cooling at 4 °C for 24, 48, or 72 h (Experiment 1); as fragments, programmed slow-freezing with dimethyl sulfoxide (DMSO), glycerol, or ethylene glycol (Experiment 2); or solid-surface vitrification using DMSO, glycerol, or ethylene glycol, each using 5-, 15-, or 30-min cryoprotectant exposure times (Experiment 3). For testis tissue xenografting, four immunodeficient recipient mice were assigned to each protocol, and each mouse received eight grafts. Recipient mice were killed 16 wk after grafting to assess the status of graft development. Based on morphology and in vitro assessment of cell viability, cooling of testis tissue for up to 72 h maintained structural integrity, cell viability, in vivo growth, and developmental potential up to complete spermatogenesis comparable with that of fresh tissue (control). In frozen-thawed testis tissues, higher numbers of viable cells were present after programmed slow-freezing using glycerol compared with that after DMSO or ethylene glycol (P < 0.001). Among the vitrified groups, exposure to DMSO for 5 min yielded numerically higher viable cell numbers than that of other groups. Cryopreserved tissue fragments recovered after xenografting had normal spermatogenesis; germ cells advanced to round and elongated spermatids after programmed slow-freezing using glycerol, as well as after vitrification using glycerol with 5- or 15-min exposures, or using DMSO for a 5-min exposure.     

Characterization of class I integrons among Salmonella enterica serovar Enteritidis isolated from humans and poultry

A total of 84 Salmonella enterica serovar Enteritidis (S. Enteritidis) isolates, 42 of human and 42 of poultry origin, were characterized for antimicrobial resistance patterns and class I integrons. Among them, 58 (69%) S. Enteritidis were multidrug-resistant (MDR) and showed resistance to two or more antibiotic classes. By PCR assays and DNA sequencing, 50 (59.5%) S. Enteritidis isolates were found to carry class I integrons. Amplification of internal variable regions of class I integrons revealed five different arrays (0.75 kb only, 1 kb only, 1.3 kb only, both 1 and 1.2 kb, and both 1 and 1.3 kb). The integrons were further sequenced and the dfrA25 (0.75 kb), aadA1 (1 kb), aadA2 (1 kb), bla(PSE1) (1.2 kb) aadA6-orfD (1.3 kb) gene cassette arrays were identified. Ciprofloxacin minimum inhibitory concentration (MIC) values for the three isolates that showed resistance or reduced susceptibility via the disc diffusion method were 0.5-4 μg mL(-1), although only three isolates exhibited resistance to cefteriaxone (MIC: 128-256 μg mL(-1)) and four isolates were resistant to florfenicol (MIC: 32-128 μg mL(-1)). In conclusion, the high rates of multidrug-resistance and class I integrons found among S. Enteritidis isolates in humans and poultry in Tehran suggest that efforts are needed to confine the prevalence of MDR Salmonella isolates.     

A case of amniotic membrane transplantation in non-healing Nocardia asteroides keratitis

A patient with severe non-healing microscopically proven Nocardia keratitis was treated with maximal topical amikacin followed by amniotic membrane transplantation (AMT) 2 months later (single-layer epithelial side-down). Epithelial healing was achieved, but neovascularization continued. This case report indicates that AMT combined with topical antibiotic provides pain relief and allows epithelial healing in severe Nocardia keratitis.     

Antioxidant and oxidative stress changes in experimental cor pulmonale

Although right heart failure (RHF) contributes to 20% of all cardiovascular complications, most of the information available on RHF in general is based on the experiences with left heart failure. This study on RHF investigates changes in antioxidants and oxidative stress which are suggested to play a role in the transition from hypertrophy to failure. RHF subsequent to pulmonary hypertension was produced in rats by a single injection of monocrotaline (MCT, 60 mg/kg, i.p.). Based on hemodynamic, clinical and histopathologic observations, the animals were grouped in three functional stages at 1-, 2- and 6-week post-injection periods. In the 1-week group, RV pressure overload and hypertrophy, and a mild increase in antioxidant enzymes was seen. In the 2-week group, compensated HF, a significant increase in antioxidant enzymes, an increase in septal (IVS) wall thickness and leftward displacement of IVS without change in LV free wall were seen. In the 6-week group, lung and liver congestion, RVF and dilation, a decrease in antioxidant enzyme activities, increase in lipid peroxidation and severe bulging of the IVS into the left ventricle were seen. These changes in the hemodynamic, biochemical and histopathologic characteristics suggest that in early stages of MCT-induced pulmonary hypertension at 1 and 2 weeks, RV hypertrophy was accompanied by sustained hemodynamic function and an increase in antioxidant reserve. In the later stage at 6 weeks, clinical RHF was associated with abnormalities of the right heart systolic and diastolic function along with a decrease in antioxidant reserve. These biphasic changes in RV antioxidant enzymes, i.e. an increase during hypertrophy and a decrease in failure may suggest a role of oxidative stress in the pathogenesis of right ventricular dysfunction.     

The Human Neutrophil Subsets Defined by the Presence or Absence of OLFM4 Both Transmigrate into Tissue In Vivo and Give Rise to Distinct NETs In Vitro

Neutrophil heterogeneity was described decades ago, but it could not be elucidated at the time whether the existence of different neutrophil subsets had any biological relevance. It has been corroborated in recent years that neutrophil subsets, defined by differential expression of various markers, are indeed present in human blood, calling for renewed attention to this question. The expression of the granule protein olfactomedin 4 (OLFM4) has been suggested to define two such neutrophil subsets. We confirm the simultaneous presence of one OLFM4-positive and one OLFM4-negative neutrophil subpopulation as well as the localization of the protein to specific granules. In vitro, these neutrophil subsets displayed equal tendency to undergo apoptosis and phagocytose bacteria. In addition, the subpopulations were recruited equally to inflammatory sites in vivo, and this was true both in an experimental model of acute inflammation and in naturally occurring pathological joint inflammation. In line with its subcellular localization, only limited OLFM4 release was seen upon in vivo transmigration, and release through conventional degranulation required strong secretagogues. However, extracellular release of OLFM4 could be achieved upon formation of neutrophil extracellular traps (NETs) where it was detected only in a subset of the NETs. Although we were unable to demonstrate any functional differences between the OLFM4-defined subsets, our data show that different neutrophil subsets are present in inflamed tissue in vivo. Furthermore, we demonstrate NETs characterized by different markers for the first time, and our results open up for functions of OLFM4 itself in the extracellular space through exposure in NETs.     

Antioxidant and oxidative stress changes in experimental cor pulmonale

Although right heart failure (RHF) contributes to 20% of all cardiovascular complications, most of the information available on RHF in general is based on the experiences with left heart failure. This study on RHF investigates changes in antioxidants and oxidative stress which are suggested to play a role in the transition from hypertrophy to failure. RHF subsequent to pulmonary hypertension was produced in rats by a single injection of monocrotaline (MCT, 60 mg/kg, i.p.). Based on hemodynamic, clinical and histopathologic observations, the animals were grouped in three functional stages at 1-, 2- and 6-week post-injection periods. In the 1-week group, RV pressure overload and hypertrophy, and a mild increase in antioxidant enzymes was seen. In the 2-week group, compensated HF, a significant increase in antioxidant enzymes, an increase in septal (IVS) wall thickness and leftward displacement of IVS without change in LV free wall were seen. In the 6-week group, lung and liver congestion, RVF and dilation, a decrease in antioxidant enzyme activities, increase in lipid peroxidation and severe bulging of the IVS into the left ventricle were seen. These changes in the hemodynamic, biochemical and histopathologic characteristics suggest that in early stages of MCT-induced pulmonary hypertension at 1 and 2 weeks, RV hypertrophy was accompanied by sustained hemodynamic function and an increase in antioxidant reserve. In the later stage at 6 weeks, clinical RHF was associated with abnormalities of the right heart systolic and diastolic function along with a decrease in antioxidant reserve. These biphasic changes in RV antioxidant enzymes, i.e. an increase during hypertrophy and a decrease in failure may suggest a role of oxidative stress in the pathogenesis of right ventricular dysfunction.