Pattern of infection and antibiotic activity among Streptococcus agalactiae isolates from adults in Mashhad,Iran

Background:

One of the main causes of sexually transmitted diseases is group B β- hemolytic streptococci (GBS) multiplying in the genital tracts. Penicillin is the most common drug for the treatment of infections caused by these bacteria, but in patients suffering from Penicillin allergy, Erythromycin and Clindamycin are used as alternative therapeutic drugs against GBS. Recently, resistance to these drugs has been reported more often. In this study, efforts have been made to determine the prevalence and antibiotic resistance of GBS.

Methods:

Modified Christie Atkins Munch-Petersen (CAMP) test was conducted on over 2400 samples of urine and discharge taken from vagina, urethra and prostate. The drug sensitivity was performed by double disk sensitivity tests to Bacitracin, Trimethoprim, and Sulfamethoxazole and then the resistant samples were investigated by E-test to determine the minimal inhibitory concentrations (MICs) value.

Results:

Twenty-three vaginal and 10 urethral discharge, 27urine and 6 prostatic secretion samples were GBS positive. The most symbiotic microorganisms with GBS were strains of Enterococci (90%), Staphylococcus saprophyticus (25%) and Candida albicans (6%). The disk diffusion method showed 18 cases with Penicillin resistance (MIC: 1.5 mg/ml).

Conclusion:

Taken together, GBS carriers’ rate in this study was found 20.65% (8.24% men and 12.4% women). Furthermore, findings showed high-level resistance to Erythromycin and Clindamycin.Key Words: Antibiotic resistance, Genitourinary system, Streptococcus agalactiae, minimal inhibitory concentration (MIC)     

Health risk assessment of fluoride in water distribution network of Mashhad,Iran

High and low levels of fluoride in drinking waters have been considered as a major public health issue in recent years. This cross-sectional study was conducted over five consecutive years (from 2012 to 2016) in the water distribution network of Mashhad city, Iran with the objectives of determining levels of fluoride and to perform health risk assessment among residents in the study area. Water samples were taken from 30 stations and were analyzed using UV-visible spectrophotometer. The mean annual concentrations of fluoride in all stations during five years of consecutive study were lower than the respective maximum permissible limits (1.5 mg/L) in water as regulated by the WHO. The human health risk assessment was performed by calculating the chronic daily intake and hazard quotient (HQ) of fluoride through human oral intake for adults (men and women) and children for each year during a five-year study. Health risk analysis in this study presented that the non-carcinogenic risk associated with fluoride exposure through drinking water in Mashhad was mostly acceptable because the mean HQ values of fluoride were lower than 1.     

Prevalence of oral mucosal lesions in institutionalized elderly people in Mashhad, Northeast Iran

doi: 10.1111/j.1741‐2358.2011.00588.x
Prevalence of oral mucosal lesions in institutionalized elderly people in Mashhad, Northeast Iran Objective: The aim of this study was to determine the prevalence of oral mucosal lesions in institutionalized elderly people in Mashhad, northeast of Iran. Background: This study was conducted to assess the prevalence of oral lesions in the institutionalized elderly. Methods: In this survey, we studied all of the nursing homes in Mashhad; a total of five encompassing 237 residents. The subjects were examined and lesions recorded in a designed checklist. T‐test, chi‐square and Fisher’s exact tests were used for the analysis. Results: The mean age of the patients was 79.59 ± 8.88. Ninety‐eight per cent of cases had at least one oral mucosal lesion. The most common lesions were fissured tongue (66.5%), atrophic glossitis (48.8%), sublingual varicosity (42%) and xerostomia (38%). There was no significant difference in the prevalence of denture‐related lesions (DRLs) between men and women (p > 0.05). Xerostomia was more prevalent in 70–79‐year‐old than in 60–69‐year‐old subjects. No case of oral malignant lesion was detected. Discussion: These findings revealed a higher prevalence of oral mucosal lesions in Mashhad city compared with other studies conducted in other major cities in Iran and abroad. The results emphasise the necessity of national programmes towards oral health promotion.     

An abattoir-based study on the prevalence of hydatidosis in livestock in Mashhad, Iran

A 6-year (2004-2010) retrospective study was carried out to determine the prevalence of hydatidosis in livestock slaughtered in Mashhad abattoir, Mashhad, in north-eastern Iran and the capital of Khorasan province. Between 20 March 2004 and 19 March 2010, 5,131,485 animals (411,163 cattle; 4,547,618 sheep; 172,704 goats) were slaughtered in the study area and 388,399 (7.5%) livers and 1,139,529 (22.2%) lungs were discarded. Hydatidosis was responsible for 4% and 6.5% of total livers and lungs inspected, respectively. Of the cattle livers and lungs inspected over the survey period, 5.5% and 7.9% were condemned, respectively, because they held hydatid cysts. The corresponding values for livers and lungs of sheep (2%, 4%) and of goats (4.5%, 7.8%), respectively, were also condemned due to hydatidosis. Data showed a prominent seasonal pattern for hydatidosis. Liver condemnations due to hydatidosis were higher in winter and autumn for cattle and sheep, respectively, whereas lung condemnations were higher in summer for sheep and cattle. In goats, liver and lung condemnations were higher in winter. This could be attributed to various factors, such as sources of slaughtered animals, changes in management practice and ecological factors. The present survey provides baseline data for the future monitoring of this potentially important parasitic disease in the region.     

An abattoir-based study of hydatidosis in the dromedary (Camelus dromedarius) in Mashhad, Iran

A 6-year retrospective study based on abattoir records was carried out to determine the prevalence of hydatidosis in dromedaries in Khorasan province in north-eastern Iran. Between 20 March 2004 and 19 March 2010, 25,255 dromedaries were slaughtered in the study area and the livers of 2791 (11.1%) and the lungs of 3289 dromedaries (13.2%) were discarded due to hydatidosis. The annual prevalence of liver condemnations due to hydatidosis decreased from 24.1% in 2004-2005 to 13.3% in 2009, and finally to 6.8% in 2010. The corresponding features for lung condemnation due to hydatidosis were relatively higher than liver, declining from 28.7% in 2004-2005 to 14.9% in 2009, and finally to 7.1% in 2010. Liver and lung condemnations due to hydatidosis were significantly higher in the spring. This could be attributed to various factors such as sources of slaughtered animals, changes in management practice and ecological factors. The present survey provides baseline data for the future monitoring of this potentially important parasitic disease in the region.     

Microtubule organization in lymphoid malignancies.

The abnormal organization of actin-containing microfilaments and vimentin-containing intermediate filaments in neoplastic lymphocytes of T and B cell origin has been described. We investigated microtubules of pathologic cells from 34 lymphoid malignancies, by immunofluorescence microscopy, using monoclonal tubulin antibody. In most cases, apart from two cases of lymphoma, one T cell lymphoma and one B cell lymphoma, interphase leukemia cells, lymphoma cells, and myeloma cells were shown to contain well-organized microtubules which were associated with a microtubule organization center at one end. In the cells of a patient with T cell lymphoma, although microtubules were not visible in the lymphoma cells from lymph nodes, they became visible after 72 hours in culture with concanavalin A (Con A) and interferon alpha. Cap formation was observed with antitubulin monoclonal antibody in the peripheral blood lymphocytes from a chronic lymphocytic leukemia patient, but well-developed microtubules were observed on other occasions in the same patient. There were no obvious structural differences between microtubules in T and B cell lymphoid malignancies, but leukemia cells and lymphoma cells with irregularly shaped nuclei, such as adult T cell leukemia cells and B cell lymphoma cells with cleaved nuclei, had complicated microtubules surrounding their irregular nuclei. In general, after blastogenic stimuli with phytohemagglutinin-P (PHA-P), Con A, and pokeweed mitogen (PWM), the development of the microtubules was proportional to the incorporation of 3H thymidine (3H-TDR). In most cases, after incubation with granulocyte colony-stimulating factor (G-CSF) and interferon alpha, the number of intact cells decreased and the number of degenerated cells increased, but the intact cells had intact microtubules.     

Fecal and Molecular Survey of Neospora caninum in Farm and Household Dogs in Mashhad Area, Khorasan Province, Iran

Neospora caninum is an important cause of abortion in dairy cattle worldwide. Dog is the definitive host for N. caninum and can infect dairy cattle. The aim of this study is to determine the prevalence of Neospora oocysts in feces of dogs from dairy farms. A total of 174 fecal samples was collected from 89 farm dogs and 85 household dogs during 2006 and 2008. Fecal samples of dogs were microscopically examined for detecting Hammondia Neospora-like oocysts (HNLO) by Mini Parasep®SF fecal parasite concentrator. HNLO were microscopically detected in 4 fecal samples (2.2%). The fecal samples with HNLO were examined by N. caninum-specific PCR. Two of the samples were positive for N. caninum. The 2 positive fecal samples were selected for inoculation to calves. Two inoculated calves were seronegative by ELISA for 4 months post-infection. This is the first report of finding N. caninum DNA in feces of farm dogs in Mashhad area, Iran.     

Surveillance of Dermatophytosis in Northeast of Iran (Mashhad) and Review of Published Studies

Dermatophytoses are considered to be one of the major public health problems in the world and are among the most commonly diagnosed skin diseases in Iran. In spite of improved personal hygiene and living environment, dermatophytosis continues to spread and persist. To determine the prevalence of dermatophytosis and their etiologic agents in Mashhad (Iran), five hundred and sixty patients suspected to have fungal infection were studied. Subjects who participated in this study were 330 males and 230 females ranged in age from 4 months to 70 years with a mean age of about 25.5 years. Clinical materials including skin scraping, hair and scalp sample, nail clipping and subungual debris were collected. All of the specimens were assessed by direct examination and culture. Of 560 patients, 166 (29.6 %) had dermatophytosis. The types of tinea according to anatomical areas were as follows: tinea corporis (33.1 %), tinea capitis (32.5 %), tinea manuum (17.5 %), tinea cruris (10.2 %), tinea pedis (5.4 %), tinea unguium (0.6 %) and tinea barbae (0.6 %). Trichophyton verrucosum was the most prevalent species followed by Trichophyton violaceum and Trichophyton mentagrophytes. The prevalence of dermatophytosis in males was higher than in females. Based on a review of published studies from different parts of Iran, there are regional differences in the incidence of dermatophytosis. Epidermophyton floccosum has been the most prevalent species, and Microsporum canis has been isolated less than from the other countries.     

Monoclonal antibodies in the treatment of lymphoid malignancies

Rituximab (Rit) was the first monoclonal antibody approved for therapeutic use in cancer patients. Rit is a chimeric mouse/human monoclonal antibody, consisting of the human IgG1 and k constant Fc region, and a mouse variable Fab region specific against the B-cell antigen CD20. Rit exerts its antilymphoma activity through many different mechanisms. Binding of antibody to CD20 antigen, provokes apoptosis through downstream signals that lead to caspase-3 activation. Complement activation by the Fc portion of the antibody results in complement-dependent cytotoxicity. However, the most effective mechanism of action seems to be antigen-dependent cellular cytotoxicity. Effector cytotoxic cells such as natural killer cells (NK) are activated after binding to the Fc portion of the anti-CD20 molecule. Activated NK cells kill the coated lymphoma cells with the use of granzyme-perforin system. More recently, pre-clinical data support the concept that Rituximab can provoke a vaccination-like effect. Finally in-vitro experiments and clinical trials have shown that co-administration of the antibody with cytotoxics confers a strong synergistic effect. The relative contribution of these mechanisms in vivo and in different lymphoma subtypes is not well known and remains to be further evaluated.

Among the different histological groups, follicular lymphoma (FL) has been proven to be the most sensitive to Rit when used as a single agent, with overall response rates of 80% and 50% in untreated and previously treated patients, respectively. Moreover, Rit in combination with chemotherapy is superior to chemotherapy alone in terms of response rate and event-free survival, while early data indicate a significant prolongation in overall survival as well. Similarly, the addition of Rit to standard chemotherapy improves the disease-free and overall survival of patients with diffuse large B-cell lymphoma. There is no doubt that Rit represents one of the greatest achievements of biotechnology engineering. However, we need to understand better the mechanisms of its action as well as the mechanisms of resistance to Rit, in order to design more effective treatment modalities.     

TNFR-associated factor family protein expression in normal tissues and lymphoid malignancies

TNFR-associated factors (TRAFs) constitute a family of adapter proteins that associate with particular TNF family receptors. Humans and mice contain six TRAF genes, but little is known about their in vivo expression at the single cell level. The in vivo locations of TRAF1, TRAF2, TRAF5, and TRAF6 were determined in human and mouse tissues by immunohistochemistry. Striking diversity was observed in the patterns of immunostaining obtained for each TRAF family protein, suggesting their expression is independently regulated in a cell type-specific manner. Dynamic regulation of TRAFs was observed in cultured PBLs, where anti-CD3 Abs, mitogenic lectins, and ILs induced marked increases in the steady-state levels of TRAF1, TRAF2, TRAF5, and TRAF6. TRAF1 was also highly inducible by CD40 ligand in cultured germinal center B cells, whereas TRAF2, TRAF3, TRAF5, and TRAF6 were relatively unchanged. Analysis of 83 established human tumor cell lines by semiquantitative immunoblotting methods revealed tendencies of certain cancer types to express particular TRAFs. For example, expression of TRAF1 was highly restricted, with B cell lymphomas consistently expressing this TRAF family member. Consistent with results from tumor cell lines, immunohistochemical analysis of 232 non-Hodgkin lymphomas revealed TRAF1 overexpression in 112 (48%) cases. TRAF1 protein levels were also elevated in circulating B cell chronic lymphocytic leukemia specimens (n = 49) compared with normal peripheral blood B cells (p = 0.01), as determined by immunoblotting. These findings contribute to an improved understanding of the cell-specific roles of TRAFs in normal tissues and provide evidence of altered TRAF1 expression in lymphoid malignancies.     

Dendritic cells and T lymphocyte interactions in patients with lymphoid malignancies

Dendritic cell (DC) vaccination is an attractive approach to the treatment of patients with lymphoid tumors. To evaluate its feasibility, we have tested the functional properties of DC and T-lymphocytes in patients with treated and untreated chronic lymphocytic leukemia (CLL) and follicular lymphoma (FL). Healthy volunteers were used both as controls and as a source of cells for allogeneic mixed leukocyte reaction (MLR). In these reactions, dendritic cells from both untreated and treated patients were comparable to dendritic cells from healthy volunteers. In all the untreated patients studied, autologous dendritic cells promoted the survival and proliferation of both CD4 and CD8 lymphocytes (though the proliferation response was much better in the CD4 subset), whereas only 3 out of 5 treated patients were able to mount this response with CD4 lymphocytes and 4 out of 5 with CD8 lymphocytes. In 3 out of 5 untreated patients, pulsing of DCs with tetanus toxoid promoted a better CD4 response than was achieved with unpulsed DCs, while none of 5 treated patients had an additional response after pulsing with tetanus toxoid. None of patients studied, either treated or untreated, had a better CD8 response to pulsed DCs than to unpulsed ones. During CD4 lymphocyte proliferation, more CD4(+)CD25(hi) lymphocytes were generated in both treated and untreated patients than in healthy controls. Poor proliferation of cytotoxic cells and preferential proliferation of CD4(+)CD25(hi) T-regulatory cells in response to self and/or foreign antigens might be one of the mechanisms responsible for immunosuppression and impaired tumor surveillance in patients with lymphoid malignancies.     

Distributions of carbon in calcareous soils under different land uses in western Iran

Concentrations of Natural stable and unstable carbon in ecosystems have been used extensively to help to understand a wide range of soil processes and functions. This study was conducted to explore the effects of land use changes on different carbon fractions (F₁, F₂, F₃ and F₄), permanganate oxidizable carbon (POXC), soil organic carbon (SOC) and total organic carbon (TOC) associated with soils in calcareous soils of western Iran. Four popular land uses in the selected site including natural forest, range land, dryland farming and irrigated farming systems were employed as the basis of soil sampling. The results showed a strong relationship between land use conversion and SOC stocks changes. The greatest mean values for carbon content and the least mean values of CaCO₃ in bulk topsoil (0–15 cm) in the forest land were observed. Dryland farming had the least both active and passive pools of C in comparison with the other land uses. The positive and significant correlations was observed between SOC, Total C and POXC contents and different C fractions. Taking C and POXC pools into account, a more definitive picture of the soil C is obtained than when only total C is measured. The influence of land use changes on overall soil carbon stocks could be helpful for making management decision for farmers and policy makers in the future, for enhancing the potential of C sequestration in western Iran.     

Quantitative monitoring of lymphoid malignancies. Application and findings in chronic lymphocytic leukemia

The present study investigated the surface immunoglobulin (Ig) phenotypic pattern in 64 cases of chronic lymphocytic leukemia (CLL). The fluorescence activated cell sorter techniques were modified to provide sensitive and highly reproducible detection and quantification of the otherwise faint surface immunoglobulins on the cells in CLL. In over 98% of the cases of CLL, light chain of the surface Ig could be identified and measured. Serial measurements were shown to be highly reproducible. The phenotypic pattern and density identified on the surface of the circulating lymphocytes precisely reflected the findings in any given patient when other lymphoid tissue (bone marrow, lymph node or spleen) was sampled. Intraclonal heterogeneity detected by surface Ig was seen in some cases of CLL in spite of a relatively uniform morphology by other classical techniques. Three patterns of cell-to-cell variation were seen: 1) that of a non-Gaussian distribution of surface light Ig staining intensity 2) that of the presence of increased surface light Ig chain on a portion of the cells with a subpopulation of CLL cells showing an additional class of heavy chain, and 3) that of anisotropy where the surface Ig quantitatively did not correlate with cell size. Immunoglobulin phenotypic characterization of the cases of CLL was correlated with their clinical stage of disease activity. The distribution of surface light chain phenotype did not relate to any pattern of clinical stage of activity of the disease. By contrast, cases where the cells had a predominance of surface IgM were associated with a more advanced stage of CLL and a poorer clinical prognosis. When surface IgG was predominant, a lesser degree of clinical activity of disease was identified. The phenotypic pattern of the surface Ig on the lymphocytes in CLL mirrors the pattern of differentiation in the murine model of B-cell differentiation, and clinical aggressiveness appears to correlate with the character and degree of B-cell differentiation.     

Microscopic anatomy of small-intestinal lymphoid patches in children and adults

Longitudinal histological sections of lymphoid patches (LP) in the small intestine, stained after van Gieson and with hematoxylin--eosin, obtained from 105 corpses of persons at the age of 0 up to 90 years have been studied. The LP parenchyma is presented as accumulation of lymphoid nodules++ with a germinative center, or without it and diffuse lymphoid tissue. The lymphoid nodules++ situate both in the mucous membrane and in the submucous tela of the small intestine. The area of the LP components in the newborns is the greatest--84% from the whole area of the LP section. During the human life it decreases and by the old age it makes 47%. The lymphoid nodules++ area with the germinative center in the newborns makes 21% from the area of the LP section, in children from 1 to 3 years of age it increases up to 50%, then it gradually decreases and in the elderly age it makes only 3.8%. The diffuse lymphoid tissue in the LP composition is revealed in all age periods. Size of the lymphoid nodules++ patches, situating in the jejunum and ileum walls in the newborns is the same. In children at the age of 10 days--12 years the LP nodules++ in the ileum are larger than those in the sejunum. The size of the LP noduli increase up to the period of the second childhood then decreases. The height of the lymphoid noduli in persons of mature, elderly and old ages decreases twice in comparison with the children of 3-12 years of age.     

Cooperative miRNA-dependent PTEN regulation drives resistance to BTK inhibition in B-cell lymphoid malignancies

Aberrant microRNA (miR) expression plays an important role in pathogenesis of different types of cancers, including B-cell lymphoid malignancies and in the development of chemo-sensitivity or -resistance in chronic lymphocytic leukemia (CLL) as well as diffuse large B-cell lymphoma (DLBCL). Ibrutinib is a first-in class, oral, covalent Bruton’s tyrosine kinase (BTK) inhibitor (BTKi) that has shown impressive clinical activity, yet many ibrutinib-treated patients relapse or develop resistance over time. We have reported that acquired resistance to ibrutinib is associated with downregulation of tumor suppressor protein PTEN and activation of the PI3K/AKT pathway. Yet how PTEN mediates chemoresistance in B-cell malignancies is not clear. We now show that the BTKi ibrutinib and a second-generation compound, acalabrutinib downregulate miRNAs located in the 14q32 miRNA cluster region, including miR-494, miR-495, and miR-543. BTKi-resistant CLL and DLBCL cells had striking overexpression of miR-494, miR-495, miR-543, and reduced PTEN expression, indicating further regulation of the PI3K/AKT/mTOR pathway in acquired BTKi resistance. Additionally, unlike ibrutinib-sensitive CLL patient samples, those with resistance to ibrutinib treatment, demonstrated upregulation of 14q32 cluster miRNAs, including miR-494, miR-495, and miR-543 and decreased pten mRNA expression. Luciferase reporter gene assay showed that miR-494 directly targeted and suppressed PTEN expression by recognizing two conserved binding sites in the PTEN 3′-UTR, and subsequently activated AKT^Ser473. Importantly, overexpression of a miR-494 mimic abrogated both PTEN mRNA and protein levels, further indicating regulation of apoptosis by PTEN/AKT/mTOR. Conversely, overexpression of a miR-494 inhibitor in BTKi-resistant cells restored PTEN mRNA and protein levels, thereby sensitizing cells to BTKi-induced apoptosis. Inhibition of miR-494 and miR-495 sensitized cells by cooperative targeting of pten, with additional miRNAs in the 14q32 cluster that target pten able to contribute to its regulation. Therefore, targeting 14q32 cluster miRNAs may have therapeutic value in acquired BTK-resistant patients via regulation of the PTEN/AKT/mTOR signaling axis.Subject terms: Cancer, Chronic lymphocytic leukaemia     

Bone marrow as a major lymphoid organ in Rana.

The bone marrow of adult frogs (Rana pipiens and R. catesbeiana) contains leukocytes equivalent in number to those present in the spleen. As in the spleen, a high percentage of these cells are morphologically identified as lymphocytes, and the bulk of these can be categorized as small or medium lymphocytes. In young frogs (R. catesbeiana), the ratio of marrow to spleen leukocytes is considerably greater than in adults, and in limited comparisons a high percentage of lymphocytes are found. A comparison of adult marrow and spleen anti-TNP (trinitrophenyl) plaque-forming cell responses at various days early in the response indicates that in the majority of animals marrow normally produces a larger total primary response.