Livestock infected with Leishmania spp. in southern Iran.

BACKGROUND: Query ID="Q1" Text="Graphical abstract: As per journal requirements, graphical abstract is necessary. Kindly check and provide the same."The magnitude of the health problems caused by leishmaniasis has been a major driving factor behind the development and implementation of leishmaniasis control programs by the national authorities in Iran, with a priority for health and environmental management. Such programs are not achievable unless all of the factors leading to the infection, including the parasite's life-cycle, vectors and reservoirs, are recognized. So far in Iran, humans and rodents have been considered the principal reservoirs of Leishmania tropica and Leishmania major, respectively, both associated with cutaneous leishmaniasis (CL), with domestic dogs considered to be the main reservoir for Leishmania infantum, associated with visceral leishmaniasis (VL). The role of other mammals in maintaining the Leishmania parasite has remained unclear. This study aimed to investigate Leishmania infection among livestock in endemic areas of VL and CL in Fars province, southern Iran, using serological and molecular methods. METHODS: Blood samples from 181 clinically healthy livestock, including 49 sheep, 114 goats, 16 cattle and two donkeys, were screened to detect Leishmania DNA and anti-Leishmania antibodies using qPCR (quantitative PCR) and the direct agglutination test (DAT), respectively. Four qPCR-positive samples were amplified using the internal transcribed spacer one (ITS1) primers in conventional PCR and sent for directional sequencing. RESULTS: Of the 181 livestock tested, 51 (28.2%) were infected with Leishmania, using serological and molecular methods. Anti-Leishmania antibodies were detected in 70 (38.7%) (95% confidence interval [CI]: 31.5-46.2) and Leishmania DNA in 93 (51.4%) (95% CI: 43.9-58.9) livestock. The identified Leishmania spp. were L. infantum and L. major. CONCLUSIONS: The findings of the present study show a relatively high prevalence of Leishmania infection among livestock in endemic areas of the disease, in Fars province, southern Iran. Given the large population of this group of animals and the fact that they live in the vicinity of the main reservoirs of the disease and vectors, it seems that sand flies regularly bite these animals. Further studies are needed to determine the role of livestock in the parasite's life-cycle and the epidemiology of Leishmania infection.

An Update of Helicobacter pylori Prevalence and Associated Risk Factors in Southern Iran: A Population-Based Study.

BACKGROUND: The childhood period is considered to be the primary period for acquisition of the Helicobacter pylori. The high prevalence rates from developing countries are associated with gastric cancer. A decreasing trend of its prevalence has been reported from different parts of the world. Determining the prevalence rate could be important in choosing preventive strategies. This study aimed to determine the prevalence of H. pylori among a group of children from southern Iran to provide an update on the current status of the disease. METHODS: This is a cross-sectional population-based study conducted in Shiraz, southern Iran, from January 2014 to December 2015. Four groups including neonates, children aged 6 months to 3 years, 10- and 15-year-old children were included. Multi-monoclonal stool antibody test was used for diagnosis. RESULTS: Among 436 participants, 24.8% (95% CI: 20.8-29.1) had a positive test for H. pylori: 25% in neonates (95% CI: 16.2-36.1), 22% in children aged 6 months to 3 years (95% CI: 15.2-30.2), 19.5% in the 10-year-old (95% CI: 12.3-29.4), and 29.2% in 15-year-old children (95% CI: 21-39). Sex, age, number of siblings, owning a pet, parents' smoking status, parental education, residential area, birth weight, and feeding status were not found to be statistically significant predictors of H. pylori antigen positivity (P > 0.05). CONCLUSION: The prevalence of H. pylori was estimated to be low in southern Iran in comparison with previous reports or other developing countries. Preventive strategies with respect to low prevalence rates may be considered in the childhood period.

Significance of TC9 and TH9 in Helicobacter pylori-induced gastritis.

BACKGROUND: H pylori plays a critical role in the development of stomach cancer, especially in people affected by the bacteria at an early stage of life. Th9 cells and IL-9 play major roles in immune responses against various infections. IL-9 is influential in chronic or acute inflammation of the mucosa. AIM: This study seeks to investigate the possible functions of Tc9, Th9 cells, and IL-9 level in patients with inflammation due to H pylori infection. METHODS: Eighty-three patients with dyspepsia symptoms and twenty normal subjects with no sign and symptoms of dyspepsia were recruited. Frequencies of T-cell subsets were determined by flow cytometry. Levels of cytokines IL-9 family in the sera and supernatants of antigen-activated PBMCs patients were measured by ELISA and flow cytometry. RESULTS: The participants included 56 females and 47 males with a mean age of 39.2 ± 15.3 years. We assigned the infected group into peptic ulcer and gastritis (chronic active and chronic). Frequencies of Tc9, Th17, Tc17, Th17/9, and Tc17/9 increased significantly in the peptic ulcer, chronic active, and chronic gastritis, compared with the uninfected and healthy control groups. A significant increase was seen in IL-9, IL-4, and IL-23 in the chronic active gastritis. Further observed was a significant increase in IL-21 and a decrease in IL-10 in the infected groups. CONCLUSION: The results revealed that increased Tc9, Th17/9, and Tc17/9 cells appear to be influential in the progression and severity of H pylori infection. Also, increased IL-9 and IL-4 levels and Tc9, Tc17/9, and Th17/9 were seen in chronic active gastritis patients. These findings may provide useful information for a therapeutic targeting of chronic active H pylori infections.

Prevalence of Chlamydia trachomatis and Neisseria gonorrhoeae infections among pregnant women and eye colonization of their neonates at birth time, Shiraz, Southern Iran.

BACKGROUND: Chlamydia trachomatis and Neisseria gonorrhoeae are the two common transmissible pathogens from pregnant women to their neonates. Given the lack of routine screening and treatment of pregnant women in some areas, the possibility of transmission rises. This study seeks to determine the prevalence of C. trachomatis and N. gonorrhoeae in the pregnant women with no clinical symptoms and the vertical transmission rate to their neonates. METHODS: The study was conducted on endocervical and eye swab samples of 239 pregnant women and their neonates. Identification was based on PCR method. RESULTS: The prevalence rates of C.trachomatis in women and neonates were 37/239 (15.5%) and 28/239 (11.7%), and for N. gonorrhoeae 3/239 (1.3%), 1/239 (0.4%), respectively. The vertical transmission rates to the neonates were 28/37(75.6%) for C. trachomatis and 1/3 for N. gonorrhoeae. CONCLUSIONS: In the areas with a high prevalence of chlamydial or gonococcal infections, and in the absence of screening and treatment of the pregnant women, ocular prophylaxis with antibiotics is suggested as a part of routine neonatal care program for the prevention of chlamydial and gonococcal ophthalmia.

Analysis of hospital records of children with hydatid cyst in south of Iran.

The clinical manifestations of hydatidosis are various and related to anatomic location. Defining frequent symptoms and signs of the disease is imperative for early management of it. The aim of this report was to analyse the clinical features of infected children with hydatid cysts located in different organs. In this study, medical charts of 57 children between 3 and 16 years of age with hydatid cyst admitted to Pediatric Wards of Nemazee Hospital were evaluated over a 12 year period (from 2003 to 2014, prospectively). All the epidemiologic, clinical, paraclinical and therapeutic data were collected. The frequencies of hydatidosis in males and females were 42.1 and 56.1%, respectively. Hydatid cysts were found in the liver and lungs in 59.6 and 33.3% patients respectively and 2 patients had an asymptomatic cyst in the heart with concomitant liver and lung cysts. The right upper quadrant pain (100%) was the most common symptom in the liver cysts. Phlegm (78.9%), Dyspnea (57.9%), acute (47.4%) and chronic cough (47.4%) were mostly seen in lung hydatid cysts. Some symptoms such as fever (68.4%) and weakness (59.6%) were the most common presenting symptoms in both groups. All children were treated through surgical approaches plus medical treatment. In the present report, liver was the most common site of involvement in children. Liver hydatidosis should be considered in children with upper quadrant pain and pulmonary hydatidosis in children complaining of phlegm and dyspnoea.

Role of serum level and genetic variation of IL-28B in interferon responsiveness and advanced liver disease in chronic hepatitis C patients.

Interleukin-28B (IL-28B) is suspected to be associated with response to treatment and one of the basic immunological backgrounds in liver transplant candidate (LTC). We aimed to assess whether genotypes of IL-28B can play a role in therapeutic response or advanced stages of liver disease. A total of 364 subjects were genotyped for IL-28B rs12979860 and rs8099917 SNPs using PCR-RFLP assay. Moreover, IL-28 serum level, HCV loads, and genotype were performed. A significant increase was observed in the frequencies of unfavorable rs12979860 genotypes/CT + TT in the chronic hepatitis C (CHC) and LTC groups. In the case of rs8099917, CHC group had a significantly higher frequency of unfavorable genotypes/GT + GG compared to the healthy group. IL-28B serum level was also significantly higher in healthy group compared with the CHC and LTC groups. There were no differences in the distribution of the IL-28B genotypes and haplotypes between responder and non-responder patients. Our results suggest, for the first time, that unfavorable rs12979860 genotypes can be considered one of the important immunological backgrounds in the Iranian LTC population that was confirmed with the lower IL-28 serum level compared to healthy group. Besides, there was a possible association of favorable IL-28B genotypes with lower odds of susceptibility to CHC infection but no support for a positive association between analyzed SNPs and an outcome of therapy. Moreover, non-CT haplotypes may be regarded as a genetic risk factor that can increase the chance of infection with HCV and progression toward end-stage HCV-related liver disease.

Conjunctival leishmaniasis in a case of disseminated cutaneous leishmaniasis.

A 15-year-old female patient presented with numerous, small, papulonodular skin lesions, and hepatosplenomegaly 9 months after a treated biopsy proved cutaneous leishmaniasis. In ocular examination there were two yellowish, raised gelatinous conjunctival lesions in the left eye. The exisional conjunctival lesion biopsy revealed many Leishman bodies inside tissue histiocytes. The patient had no systemic immunologic problems (normal serum immunoglobulins, nitroblue-tetrazolium test, complement CH50 test and flow cytometry of leukocytes). The indirect immunofluorescent antibody test for Leishmania tropica (titre of 1:1024) and the leishmanin skin test were positive. DNA of L. tropica was detected by a specific polymerase chain reaction on whole blood, bone marrow and skin biopsy specimens. The skin and conjunctival lesions disappeared with miltefosine and no intraocular tissue penetration of organism happened. Conjunctival leishmaniasis should be considered in the differential diagnosis of raised conjunctival lesions in a disseminated cutaneous leishmaniasis patient and needs proper systemic therapy.

A 25-year surveillance of disseminated Bacillus Calmette-Guérin disease treatment in children in Southern Iran.

Disseminated Bacillus Calmette-Guérin (BCG) disease is one of the most serious complications of BCG vaccination, mainly among immunocompromised children with high morbidity and mortality.Currently, there is no any consensus with regard to the standard regimen of antituberculosis (anti-TB) agents and duration of treatment in healthy or immunocompromised host in children. The aim of this study is to investigate the effect of various combination treatment strategies for disseminated BCG disease in children.In this cross-sectional study, the outcome of 3 different combination protocols was investigated in 59 patients.All patients were younger than 6 years old. Both possible immunocompetent and proven immunodeficient children were included in a period of 25 years (1991-2014) in a Nemazee referral teaching hospital.The minimum age was 1 month and the maximum was 60 months. The average age of patients was 8 months (8.26 ±â€Š9.73). Out of 59 cases, 32 (54.2%) were female and 27 (45.8%) were male. Based on the primary work up, 52.5% of cases were classified as definite immunodeficient and 47.5% were classified as possible immunocompetent. Overall mortality rate was 50.8%. Mortality rate of disseminated BCG disease in immunocompetent and immunodeficient children was 28.6% and 71%, respectively. The mortality rate was not statistically different between patients treated with different treatment protocols. These results were not affected by immune status and the type of immunodeficiency.More than 2 anti-TB drugs combination will not change outcome of patient with disseminated BCG disease.

The importance of TH22 and TC22 cells in the pathogenesis of Helicobacter pylori-associated gastric diseases.

BACKGROUND: An association exists between Helicobacter pylori (H. pylori), peptic ulcers, gastritis, and sometimes gastric carcinomas. Th22 cells have protective and inflammatory roles in defense against microbes. AIM: We investigated the frequencies of Th22, Tc22, Th22/17, and Tc22/17 cells in addition to the changes in levels of cytokines IL-22, IL-6, IL-23, TNF-α, IL-1ß, and TGF-ß in sera from patients with H. pylori-associated gastritis, and peptic ulcer, and in uninfected patients. METHODS: A total of 76 patients with H. pylori-associated disorders formed the studied group. Frequencies of T-cell subsets were determined by flow cytometry. Levels of cytokines IL-22, IL-6, IL-23, TNF-α, IL-1ß, and TGF-ß in the sera and supernatants of patients were measured by ELISA and flow cytometry. RESULTS: The study participants included 32 males and 44 females with a mean age of 38.5±15.3 years. We divided the infected group into peptic ulcer and gastritis (mild, moderate, active chronic, and chronic). The frequencies of Th22, Tc22, and Tc22/17 increased significantly in the peptic ulcer, moderate, active chronic, and chronic gastritis groups compared to the uninfected group. Th22/17 only increased significantly in the chronic gastritis group. We observed significant increases in IL-22 in the moderate and active chronic gastritis, IL-23 in the active chronic and chronic gastritis, and TNF-α in the peptic ulcer and moderate gastritis groups. Following in vitro antigenic stimulation, we observed significantly higher levels of IL-1ß, IL-23, and IL-6 in the active chronic gastritis group, as well as IL-6 and IL-1ß in the chronic gastritis group compared to the uninfected group. CONCLUSION: Increased Th22, Tc22, and Tc22/17 cells and IL-22 levels appear to be influential in progression and severity of H. pylori infection. Th22/17 can be an interesting therapeutic target for chronic H. pylori infections where eradication is more difficult.

Effectiveness of Short-Course Meglumine Antimoniate (Glucantime®) for Treatment of Visceral Leishmaniasis: A 13-Year, Multistage, Non-Inferiority Study in Iran.

The World Health Organization's (WHO) recommendation is 28-day course of meglumine antimoniate (Glucantime®, Sanofi Aventis, France) for the treatment of visceral leishmaniasis (VL). The aim of this study was to evaluate the effectiveness of a shorter duration of treatment in regions with low level of resistance to Glucantime. During 13 years, this study was conducted in three phases on 392 patients. In the pilot first phase, we performed splenic punctures in seven patients to assess the correlation between the changes in the parasite load during treatment with Glucantime and defervescence. With defervescence, parasite density was dramatically dropped (P = 0.014), propounding defervescence as a marker of parasitological response. On the basis of the results, we conducted a randomized trial on 75 patients, comparing the efficacy of continuation of Glucantime therapy for 1, 2, or 3 weeks after defervescence. The treatment course of 1 week after defervescence (mean = 11.7 days) was non-inferior to that of 3 weeks (final cure rate, 96% versus 100%; P = 0.023). The third phase was a retrospective cohort study of 302 patients treated either with the WHO's regimen or for 7 days after defervescence (intervention group). Relapse was detected in 8.3% patients of the intervention group and in 5% patients following the WHO's regimen (P = 0.006 for non-inferiority). The final duration of treatment in intervention group was significantly shorter than standard course (13.3 ± 2.6 versus 28 days; P < 0.001). In summary, treatment of VL with Glucantime for 1 week after defervescence was non-inferior to and appears to be an acceptable alternative to the standard 28-day course for patients in Iran who show a response to antimonial therapy.

Visceral leishmaniasis in two patients with IL-12p40 and IL-12Rß1 deficiencies.

Mutations of the IL12B and IL12RB1 genes underlie the development of IL-12 p40 and IL-12Rß1 deficiencies, respectively, both of which cause predisposition to infection with weakly virulent mycobacteria and Salmonella. Infections with other intramacrophagic organisms have only been rarely observed. We identified two patients with visceral leishmaniasis who had autosomal recessive IL-12 p40 and IL-12Rß1 deficiencies, respectively. This finding demonstrates the importance of IFN-γ immunity in the control of leishmaniasis. We also searched the literature for similar reports in patients with these and other primary immunodeficiencies.

A diagnostic one-step real-time reverse transcription polymerase chain reaction method for accurate detection of influenza virus type A.

INTRODUCTION: Influenza A is known as a public health concern worldwide. In this study, a novel one-step real-time reverse transcription polymerase chain reaction (rtRT-PCR) assay was designed and optimized for the detection of influenza A viruses. MATERIAL AND METHODS: The primers and probe were designed based on the analysis of 90 matrix nucleotide sequence data of influenza type A subtypes from the GenBank database of the National Center for Biotechnology Information (NCBI). The influenza virus A/Tehran/5652/2010 (H1N1 pdm09) was used as a reference. The rtRT-PCR assay was optimized, compared with that of the World Health Organization (WHO), and its analytical sensitivity, specificity and reproducibility were evaluated. In total, 64 nasopharyngeal swabs from patients with influenza-like illness (ILI) and 41 samples without ILI symptoms were tested for the virus, using conventional cell culture, direct immunofluorescence antibody (DFA) methods, and one-step rtRT-PCR with the designed primer set and probe and the WHO's. RESULTS: The optimized assay results were similar to the WHO's. The optimized assay results were similar to WHO's, with non-significant differences for 10-103 copies of viral RNA/reaction (p > 0.05). It detected 10 copies of viral RNA/reaction with high reproducibility and no cross reactivity with other respiratory viruses. A specific cytopathic effect was observed in 6/64 (9.37%) of the ILI group using conventional culture and DFA staining methods; however, it was not seen in non-ILI. Also, the results of our assay and the WHO's were similar to those of viral isolation and DFA staining. CONCLUSIONS: Given the high specificity, sensitivity and reproducibility of this novel assay, it can serve as a reliable diagnostic tool for the detection of influenza A viruses in clinical specimens and lab experiments.

Immunization with a Recombinant Expression Vector Encoding NS3/NS4A of Hepatitis C Virus Genotype 3a Elicits Cell-Mediated Immune Responses in C57BL/6 Mice.

Today, hepatitis C virus (HCV) infection is considered as one of the most significant international health concerns. Although novel therapeutic regimens against the infection have shown satisfactory results, no approved vaccine exists yet. This study aimed to evaluate the immunogenicity of a DNA vaccine candidate for HCV-3a, based on nonstructural proteins NS3/NS4A, in C57BL/6 mice. Immunogenicity effect of pDisplay-NS3/NS4A was analyzed through immunization with 100 and 200 µg concentrations of the construct with complete Freund's adjuvant, monophosphoryl lipid A (MPL), or without adjuvant. The frequencies of different splenic mononuclear cells were measured using the Mouse Th1/Th2/Th17 Phenotyping Kit. Moreover, the number of T-CD8(+) cells was determined using conjugated anti-CD8a and anti-CD3e antibodies by flow cytometry. As observed, the frequencies of Th1, T-CD8(+), and Th2 cells increased in all the experimental groups, compared with the controls. The highest levels of the respective cells were seen in the group immunized with 200 µg of the construct with MPL. Also, there were positive correlations between the frequency of Th1 cells and those of Th2 and T-CD8(+) cells in all the immunized groups, but were significant in those receiving adjuvants. The frequency of Th17 cells did not statistically change among the groups. Taken together, our findings revealed that the constructed DNA vaccine encoding HCV-3a NS3/NS4A gene induces the cell-mediated immune responses significantly. However, its coadministration with adjuvants exhibits more efficient results than the recombinant plasmid alone. Further study is currently underway to evaluate the specific immune responses and recognize the responsible antigenic epitopes.

Interference of Vaccine Derived Polio Viruses with Diagnosis of Enteroviral Diseases in Neonatal Period.

INTRODUCTION: Enteroviruses (EV) are a common cause of neonatal sepsis especially at the junction of summer and fall. AIM: This study was planned to find the frequency of Enteroviral (EV) sepsis among neonates with clinical sepsis. MATERIALS AND METHODS: This is a prospective descriptive study. Rectal and pharyngeal swab samples were taken from all neonates with clinical sepsis and a control group of neonates with simple jaundice. EV was confirmed by both cell culture and RT-PCR. Anti polio antiserum was used to differentiate Polioviruses from Non Polio EVs (NPEV). RESULTS: We had 67 neonates with clinical sepsis and 31 cases of simple jaundice during 105 days. NPEVs were isolated from 2 cases (2.9%) of the sepsis arm and one neonate (3.2%) of the jaundice group. Polio virus was isolated from 16.2% and 15.3% of OPV recipients in the sepsis and jaundice group respectively. CONCLUSION: Enteroviruses were not a common cause for neonatal sepsis in Nemazi hospital at the time of this study. OPV vaccinated neonates commonly pass the vaccine virus in their pharynx and stool which can be mistaken with NPEV.

Nosocomial Infections: Multicenter surveillance of antimicrobial resistance profile of Staphylococcus aureus and Gram negative rods isolated from blood and other sterile body fluids in Iran.

BACKGROUND AND OBJECTIVE: Antibiotic resistance is increasing, especially in healthcare-associated infections causing significant public health concerns worldwide. National information is required to make appropriate policies, update list of essential drugs for treatment, and evaluate the effects of intervention strategies. A nationwide surveillance of antimicrobial resistant bacteria in nosocomial infections was established in Iran in 2008, so that the data obtained through the surveillance would enable us to construct a database. MATERIALS AND METHODS: Seven major teaching hospitals in Shiraz, Tabriz, Sari, Mashhad, Sanandaj, Ahwaz and Isfahan participated in this study. A total of 858 strains isolated from blood and other sterile body fluids were tested. Identification at the species level was performed with conventional biochemical methods and the API system. Susceptibility tests were done using disk diffusion method. The methicillin-resistance in S. aureus (MRSA) was determined by the oxacillin agar screen plate and respective MIC values were assessed using the E-test strips. The confirmatory disk diffusion methods were applied for phenotypic identification of extended-spectrum ß- lactamase (ESBL) production for E. coli and K. pneumoniae, according to CLSI guidelines. RESULTS: Cultivation and re-identification of the strains yielded 858 isolates, consisting of 224 S. aureus, 148 Klebsiella spp., 105 Serratia spp., 146 E. coli, 67 Acinetobacter spp., 38 Enterobacter spp., 95 Pseudomonas spp., 71 P.aeruginosa. 35 Stenotrophomonas sp., and 8 other organisms. MRSA was detected in 37.5% of the isolates. No vancomycin-resistant or vancomycin-intermediate resistant S. aureus was detected. With the exception of Acinetobacter and Stenotrophomonas, 85% of the Gram-negative isolates were found to be susceptible in vitro to imipenem. Overall, about 61% of K. pneumoniae and 35% of E. coli isolates were ESBL producing. CONCLUSION: Multidrug resistant isolates of Gram-negative organisms and methicillin-resistant strains of S. aureus have been detected in many hospitals in this study.

QuantiFERON-TB Gold and Tuberculin Skin Test for the Diagnosis of Latent Tuberculosis Infection in Children.

BACKGROUND: Appropriate diagnosis and treatment of latent tuberculosis infection (LTBI) play the most important role in the control of tuberculosis. This study aimed to determine the prevalence of LTBI among healthy tuberculosis unexposed children vaccinated with BCG using the tuberculin skin test (TST) and QuantiFERON TB Gold In-Tube (QFT-GIT) and comparing the agreement between the two tests. METHODS: A cross-sectional study was carried out between October 2009 and March 2010 in 24 schools and 11 daycare centers. A total of 967 children were divided into 15 age groups, with a minimum of 64 children per group. RESULTS: The prevalence rates of LTBI with TST were 3.8%, and 2.2% with QFT-GIT. One case was positive in TST and QFT-GIT, 20 cases were QFT-GIT positive, but TST negative and 36 cases were TST positive, but QFT-GIT negative, and finally, 910 cases were negative in both. There was poor agreement between TST and QFT-GIT (1.8%, 95%, CI: 0%-5.3%, k=0.007). The specificity of QFT-GIT in the BCG vaccinated, children aged 1-15 years old, was 97.8% (97.8%, 95% CI: 96.8%-98.8%). After three months, 2/17 (11.8%) of those initially QFT-GIT negative converted, and 10/15 (66%) of those initially QFT-GIT positive reverted. CONCLUSION: It seems that TST and QFT-GIT are not appropriate tests for the diagnosis of LTBI among healthy tuberculosis unexposed BCG vaccinated children. There was a low reproducibility rate of QFT-GIT. The cause of the the poor agreement requires further studies.

Alterations in Th17 and the Respective Cytokine Levels in Helicobacter pylori-Induced Stomach Diseases.

BACKGROUND: Infections by Helicobacter can cause the stimulation of sophisticated immune response in mucosal immunity. Among the different lymphocytes, Th17 plays an important role in the defense against H. pylori and may cause gastritis and peptic ulcer due to the increased activation of Th17 and cytokine changes. AIM: To find a relationship between Th17 and IL-17A, IL-21, IL-22, IL-23, TGF-ß in the patients with H. pylori infection having signs including gastritis and peptic ulcer. METHODS: A total of 36 samples from the patients [24 Hp+ and 12 Hp- cases] with dyspepsia symptoms were collected. The percentage of Th17 was measured by flow cytometry. The levels of Th17-associated cytokines in the sera and supernatants of peripheral blood mononuclear cells (PBMCs) which were stimulated with the H. pylori antigen, phytohemagglutinin (PHA), or Dynabeads were measured by ELISA. RESULTS: Patients were divided into two groups of having either H. pylori infected (peptic ulcer, gastritis (mild, moderate)) or being uninfected. The percentage of Th17 in the patients with peptic ulcer and gastritis was significantly higher than their uninfected counterparts (p ≤ .001). The serum levels of IL-17A, IL-23, and TGF-ß in the peptic ulcer and gastritis groups were significantly higher compared with the corresponding levels in the uninfected population (p < .05). A significant association of TGF-ß, IL-21, and Th17 was observed with low levels of IL-17A in the mild gastritis patients (p < .05). Significantly higher levels of IL-22, IL-17A, IL-23, and higher Th17 frequencies were detected in the moderate gastritis patients, as compared with the uninfected patients (p ≤ .001). CONCLUSION: It can be concluded that among the cytokines associated with Th17, the two cytokines of IL-21 and TGF-ß play a more critical role in peptic ulcer and gastritis in the individuals infected with H. pylori. Furthermore, inflammation varies depending on the type of the cytokine and its secreted level.

Post-Vaccination Disseminated Bacillus Calmette Guerin Infection Among Children in Southern Iran.

BACKGROUND: Disseminated bacillus calmette guerin (BCG) infection is a rare but life threatening complication of BCG vaccination. It has been mainly seen in severe immune deficiency. A precise and rapid diagnosis is crucial for prompt initiation of an aggressive anti-mycobacterial treatment. Polymerase chain reaction (PCR) is directly applicable to smear-positive clinical specimens, proven to be a rapid and specific diagnostic test. OBJECTIVES: The aim of this study was to investigate disseminated BCG infection among 34 children in southern Iran, mainly confirmed by PCR. PATIENTS AND METHODS: We included all the patients hospitalized with disseminated BCG infection at a referral teaching hospital in southern Iran between years 1990 and 2007. The clinical and laboratory data including the immunological workups were obtained through a review of the medical files. We recalled all pathology samples from pathology specimen banks and used an in-house PCR specific for Mycobacterium bovis BCG substrain to confirm the diagnosis. RESULTS: From the total of 34 children hospitalized with disseminated BCG infection, 21 were categorized as definite and 13 probable. Thirty-one patients (91%) were under two years of age and 41% were male. The most common clinical findings were fever in 31 (91.2%), axillary's lymphadenopathy in 26 (76.5%), hepatosplenomegaly in 25 (73.5%), stunted growth in 21 (61.8%), and distant lymphadenopathy in 16 (47.1%). Polymerase Chain Reaction positivity rate was 100% (9 of 9) in bone marrow smear slides and 84.2% (16 of 19) for formalin-fixed and paraffin-embedded tissue specimens. Immunodeficiency state was detected in 50% and the overall mortality rate was 58.8% (20 of 34). CONCLUSIONS: Disseminated BCG infection should be considered in the differential diagnosis of infants and young children with fever, hepatosplenomegaly, lymphadenopathy, and history of BCG vaccination. The PCR method has a high positivity rate and can serve as a useful tool for the rapid and specific identification of M. bovis BCG substrain infection.

Expression of NS3/NS4A Proteins of Hepatitis C Virus in Huh7 Cells Following Engineering Its Eukaryotic Expression Vector.

BACKGROUND: Although the development of novel therapeutic regimens to combat hepatitis C virus (HCV) infection have been speeded up with successful results, no efficient vaccines exist yet. OBJECTIVES: This study aimed to construct a eukaryotic expression vector encoding nonstructural proteins, NS3/NS4A, of HCV genotype 3a, and evaluate its expression on Huh7 cell surface. MATERIALS AND METHODS: The NS3/NS4A sequence was isolated from a patient with HCV-3a chronic infection, cloned into intermediate vector pTZ57R/T, and then used for engineering a mammalian expression vector, pDisplay, to direct the respective protein to the secretory pathway and anchor it to the plasma membrane. The expression of the protein in Huh7 cell, which was transiently transfected with the vector using Lipofectamine, was determined by immunocytochemical staining assay with fluorescein isothiocyanate (FITC)-conjugated antibodies to the HA/myc tags located besides the fusion fragment. RESULTS: The results showed that the fragment was successfully amplified and cloned into a eukaryotic expression vector. Sequencing and enzyme digestion analysis confirmed the cloned gene completion and its correct position in the pDisply-NS3/NS4A plasmid. Immunocytochemical staining revealed that the target protein was expressed as a membrane-anchored protein in the Huh7 cells. CONCLUSIONS: This study can serve as a fundamental experiment for the construction of a NS3/NS4A eukaryotic expression vector and its expression in mammalian cells. Further research is underway to evaluate the fragment immunogenicity in lab animal models.

Yeast colonization and drug susceptibility pattern in the pediatric patients with neutropenia.

BACKGROUND: Pediatric patients with neutropenia are vulnerable to invasive Candida infections. Candida is the primary cause of fungal infections, particularly in immunosuppressed patients. Candida albicans has been the most common etiologic agent of these infections, affecting 48% of patients. OBJECTIVES: The aim of this study was to identify Candida spp. isolated from children with neutropenia and determine the antifungal susceptibility pattern of the isolated yeasts. PATIENTS AND METHODS: In this study 188 children with neutropenia were recruited, fungal surveillance cultures were carried out on nose, oropharynx, stool, and urine samples. Identification of Candida strains was performed using germ tube and chlamydospore production tests on an API 20 C AUX system. Susceptibility testing on seven antifungal agents was performed using the agar-based E-test method. RESULTS: A total of 229 yeasts were isolated. Among those, C. albicans was the most common species followed by C. krusei, C. parapsilosis, C. glabrata, C. tropicalis, C. famata, C. dubliniensis, C. kefyr, and other Candida species. C. glabrata was the most resistant isolated yeasts, which was 70% resistant to fluconazole and 50% to itraconazole, 7.5% to amphotericin B and 14% to ketoconazole. All the tested species were mostly sensitive to caspofungin. CONCLUSIONS: Knowledge about the susceptibility patterns of colonized Candida spp. can be helpful for clinicians to manage pediatric patients with neutropenia. In this study, caspofungin was the most effective antifungal agent against the colonized Candida spp. followed by conventional amphotericin B.