The Study of Uropathogenicity Factors for Escherichia Coli Strains.

Urinary tract infection (UTI) is the most common form of extraintestinal Escherichia Coli infection (E.coli), and E. coli is the most common cause of UTI.The aim of this paper is to study the uropathogenicity factors for some strains of E.coli involved in the etiology of UTI and the affiliationof urinary E.coli strains to the serogroups involved in the UTI.We studied 208 strains of E. coli from urine samples sterilely collected from patients with clinical suspicion of urinary tract infection.The study was conducted in Emergency County Hospital Craiova between 2012-2014.Out of the 208 strains of E. coli submitted to the study, 60 strains (28.84%) - MRHA with human red cells, 28 strains (13.50%) - MRHA human red cells and blood red cells MSHA with guinea pigs, and 44 strains (21.12%) - MSHA with guinea pig red blood cells; 76 strains (36.54%) - no hemagglutination. Regarding our study, 42,34% of E.coli strains presented human MRHA putting forward their potential to cause pyelonephritits. The 68 hemolytic strains (37,20%) of urinary E. coli were tested for the production of the cytotoxin, thus obtaining characteristic cytotoxic effect for 26 strains (38.20%) whereas its absence was registered in 42 strains (61.80%). E. coli O6 strains isolated from hospitalized adults are more frequently hemolytic than those isolated from the other groups and MRHA was more common in hemolytic strains of the same group O6). Mannose-resistant hemagglutination is more frequent in strains that develop HLy but do not produce CNF(Cytotoxic Necrotizing Factor), than in strains producing CNF.

Incidence and Antifungal Susceptibility of Candida Albicans Infections.

INTRODUCTION: Candida albicans is the most common inhabitant of the skin, mouth, vagina and gastro intestinal tract of human beings. One of the major reasons for the increase in Candida infection is the development of its resistant strains due to drugs used in the treatment of candidiasis. MATERIALS AND METHODS We studied 4027 samples collected from patients in various wards of the Emergency County Hospital Craiova, Romania between 2014-2015. The specimens were: pharyngeal exsudates, sputum, tracheal secretions, skin secretions, stools, ear secretions, urine, vaginal secretions. All the specimens were transported to the microbiology laboratory and cultured within 3 to 4 h of collection. Among the 4027 samples, 652 showed culture characteristics similar to Candida albicans.The samples were inoculated under sterile conditions using Sabouraud culture media, a medium designed to inhibit bacterial growth and allow the development of fungi. Antifungal Susceptibility Testing was performed by disc diffusion according to CLSI 2014 guidelines using: clotrimazole, ketoconazole, miconazole, econazole, amphotericine B, fluorocytozine, nistatin. RESULTS AND DISCUSSION In our study group the urocultures and dermatological products have a high infection rate, between 100% to 70%, in contrast, we find evidence of secretion ear (3.13%) and the throat swab (9.33%). Various resistant levels were detected against antifungal drugs but, complete resistance to 5 - Fluorocitozina (100%), and the organisms showed highly sensitive to Cotrimazol si Ketoconazol (100%). In the case of Miconazol 256 (39, 26%), Econazol, 215 (32,98%), Amphotericinei B, 230(35,28%). Nystatin 329 (50,46%). CONCLUSIONSElucidating these mechanisms may provide new foundations for antifungal chemotherapy and can present an exciting challenge for the future investigations. Candida albicans infections are present and diverse clinical pathology.

[Subconjunctival nodule with Dirofilaria repens]. / Nodul subconjunctival cu Dirofilaria repens.

Dirofilaria repens infection, a zoonotic illness, is rarely seen in humans; it is more frequently met over the endemic areas of the Southern Europe and Sri Lanka. The authors report a case of dirofilaria conjunctivae in a 27 year old woman from Craiova; such infections are hardly to be found in our country. Our patient presented a mobile, bulbar, subconjunctival tumoral formation which was surgically removed; immature Dirofilaria repens was revealed into it. Case particularity was that the patient had not visited any of the endemic areas. Environmental changes with global warming can lead to some new unspecific diseases in our country.

Current laboratory assays and in vitro intracellular Th1 and Th2 cytokine synthesis in monitoring antiretroviral therapy of pediatric HIV infection.

In 42 HIV-infected children, 8-10 years old, belonging to the category A state of infection, combined antiretroviral therapy (cART) was applied, consisting of AZT, ddC and Saquinavir. At 6 and 12 weeks following the start of cART, the efficacy of treatment was assessed, both by means of current parameters (blood CD4+ cell levels, virus load) and by measuring the intracellular synthesis of some Th1 (interleukin (IL)-2, interferon-gamma) and Th2 (IL-4, IL-10) cytokines, in CD4+ lymphocytes, respectively. Before cART, low values of blood CD4+ cell counts and a mean of about 8000 virus RNA copies ml(-1) of serum were detected, and in addition decreased levels of production of both intracellular Th1 cytokines, associated with increased levels of one of the Th2 cytokines (IL-10), but not of the other (IL-4), were noticed. After cART, earlier improvement of intracellular IL-2, interferon-gamma and IL-10 synthesis in CD4+ cells occurred compared to CD4+ counts and virus load. The usefulness of scoring the rates at which CD4+ lymphocytes are able to synthesize intracellular Th1 or Th2 cytokines, as an additional immune parameter during combined antiretroviral therapy monitoring in pediatric AIDS, is discussed.

Expression of activation surface markers, interleukin-2 synthesis and apoptosis rate in fresh or cultured lymphocytes from HIV-infected children.

In 2 groups of HIV-infected children aged from 7 to 10 years (ARC and AIDS, respectively) the following immune markers: % expression of CD69, CD25, HLA-DR activation surface determinants, IL-2 synthesis, rate of apoptosis were tested in non stimulated, PHA-stimulated or PHA and IL-12-stimulated T cell cultures. In all HIV-originated cells a decrease of CD69 expression and an increase of CD25 and HLA-DR expression were found. A strong correlation could be noticed between the clinical stage of the AIDS infection and the in vitro IL-2 production and the percentages of apoptotic cultured cells. IL-12 supplementation of PHA-stimulated cell samples restored the IL-2 synthesis and reduced the apoptosis rates only in the ARC-group, but not in the AIDS-group. The significance of the present data in the clinical and therapeutic monitoring of HIV infection among low-aged people are discussed.

Multi-way flow cytometric immune monitoring of antiretroviral therapy in HIV-infected children.

In 40 HIV-infected children, 8-10 years old, belonging to the category A state of infection, the following flow-cytometric parameters were scored: percentage levels of different blood lymphocytes; surface expression of some activation and memory markers in CD4+ cells; switch to Th1 or Th2 of in vitro -stimulated CD4+ cell, tested by intracellular production of interleukin-2 or interleukin-4. Each investigation was carried out both before and 3 months after antiretroviral therapy (AZT and ddC). Some post-therapy changes concerning blood lymphocyte percentages were noticed, not only within CD4+ subpopulation, but also within CD8+, HLA-DR+/CD3 (T-activated) and CD16+CD56+ cells, respectively. On the other hand, following antiretroviral treatment, in HIV- originated CD4+ fresh cells, an improvement of pre-therapy increased values of surface activation (CD69, CD25) markers on memory (CD45RO+) cells, as well as of pre-therapy reduced rate of switching to Th1, revealed by intracellular interleukin-2 synthesis, was found. The significance of data obtained in the multi-way immune monitoring of antiretroviral therapy, in pediatric AIDS, as an additional investigation panel, is discussed.

Different patterns of some systemic immunological cell markers in HIV only, and HIV/hepatitis C-infected children.

In three groups of children, aged 4-6 years (i.e., human immunodeficiency virus [HIV]-negative controls, HIV-seropositive, and dually HIV/hepatitis C virus (HCV)-seropositive), two types of immunological investigations in blood cells were performed: (a) numerical assays, consisting of flow cytometric measurement of different lymphocyte sets or subsets, as follows: CD3+, CD19+, CD4+, CD16+/CD56+; (b) functional assays, consisting of interleukin-1 (IL-1) levels as well as natural killer (NK)-cell dependent cytotoxicity, in CD14+, or CD16+/CD56+ sorted cells, respectively. Results revealed, in addition to the classic markers (i.e., lower numbers of CD4+ cells and a decreased CD4+/CD8+ ratio in both infected groups of subjects) other findings, as follows: increased numbers of CD8+ cells in dually infected children, accompanied by a lower CD4+/CD8+ ratio, as compared to HIV-infected alone; diminished numbers of CD16+/CD56+ cells in both groups of infected patients were correlated with a lower NK-cell cytotoxicity rate; a reduced capacity for IL-1 synthesis of sorted macrophages both in HIV-only and in HIV/HCV-seropositive subjects, but significantly more marked in dually infected children. The importance of the present data in the immune monitoring of AIDS disease in a pediatric population is discussed.

Blood amounts and suppressive in vitro activity of CD8+11b(+)-lymphocytes from HIV-infected children, related to hepatitis B or C virus association.

In HIV only, as well as in HIV/hepatitis B- and HIV/hepatitis C-infected children (6 to 8 years of age), with moderate altered clinical and immune cell markers, the total amounts of CD8(+)-set and of CD8+11b(+)-subset (T-suppressor) blood lymphocytes, by means of flow cytometry, were determined. On the other hand, within several mixtures of autologous sorted CD3+/CD8+11b(+)-cells, the per cent reduction of HLA-DR+ expression on T-cells, at different effector/responder cell ratios, was appreciated. Significant higher levels of CD8+11b(+)-cells, especially in HIV/hepatitis B virus groups, were found, that correlated with a stronger suppressive activity. The strongest alteration of immune markers, within HIV-seropositive, HBs+, HBe+ patients, was noticed. A possible usefulness of these data in HIV only, and in HIV-associated hepatitis B or C virus contamination, during pediatric AIDS monitoring, was commented.

Changes of blood CD16/CD56 (NK) and HLA-DR/CD3-positive lymphocyte amounts in HIV-infected children, as related to clinical progression and p24-antigen/p24-antibody presence.

This study describes a series of immunological investigations carried out on a group of 37 HIV-seropositive children, aged 3-4 years, in two different stages of disease defined according to the CDC classification; the Primary stage, an asymptomatic one, showing abnormal immune function (P1-Class, B-Subclass) and the Secondary stage, 6-8 months later, in which patients exhibited non-specific findings, i.e., loss of weight, persistent generalized lymphadenopathy and hepatosplenomegaly, associated with abnormal immune function (P2-Class, A-Subclass). In both stages, immune function was considered 'abnormal' when lymphopenia and a decrease of the CD4/CD8-cell ratio were found. The phenotypes CD16+/56+ (NK) and HLA-DR+/CD3+ (T-activated?)-positive cells, were assessed by flow cytometry, and the following supplementary systemic humoral markers were investigated in homologus serum samples; total HIV(gp)-antibody, HIV(p24)-antibody and p24-antigen presence. If at the primary stage, no significant difference from to the reference values corresponding to the age was noticed, at the Secondary stage the obtained data is presented separately in two subgroups, namely the A-subgroup characterized by the presence of total HIV(gp)-antibody, the presence of HIV(p24)-antibody and the absence of p24-antigenaemia, and the B-subgroup, where total HIV(gp)-antibody was present, HIV(p24)-antibody absent and p24-antigenaemia present. A significant decrease of CD16+/56+ (NK)-cells was found within the two subgroups. As far as HLA-DR+ from CD(3+)-cells was concerned, only those within the B-subgroup showed a high percentage level, compared to the reference values. The importance of the present findings, linked to immune monitoring of HIV infection among children, is discussed.

Comparative study on some systemic humoral and cellular immune markers in only HIV or HIV and hepatitis B infected children.

Three different groups of asymptomatic children, aged from 12 to 24 months (30 subjects per each group), i.e. controls, only HIV, or HIV/hepatitis B virus (HBV) double infected, were studied, as concerned the following systemic immune parameters: immunoglobulin (IgG, IgM, IgA, IgD) levels; absolute numbers of blood CD+4, CD+8, CD+16 and CD+19 cells; phytohaemagglutinin (PHA)-blast responsiveness of T lymphocytes; natural killer (NK) cell activity--as tested by means of cytotoxicity assays; per cent suppression of PHA-dependent T cell blastogenesis in the presence of concanavalin A (Con A) selected T suppressor (Ts) cells. On the other hand, in 15 ARC-shifting cases belonging to HIV, and HIV/HBV groups, respectively, a second serum sample was collected and searched comparatively with the corresponding first serum sample, as regarded: presence of total and anti-p24 HIV antibodies, patterns of Western Blot (WB), as well as amounts of free p24-HIV antigen. In asymptomatic double HIV/HBV infected subjects, some immune disorders occurred, at a more significant degree, as compared to only HIV-infected. Once the shift toward ARC being installed, in both infected groups a decrease of anti-p24 HIV antibody presence, disappearance of corresponding band in WB confirmation test, as well as presence of free p24 antigen in serum, were noticed. However, greater amounts of p24 antigen in HIV/HBV infected, as compared to only HIV infected patients, were found. Some considerations about diagnostic and predictive value of presented data are discussed.

Comparative study on some systemic humoral and cellular immune markers in only HIV or HIV and hepatitis B infected children.

Three different groups of asymptomatic children, aged from 12 to 24 months (30 subjects per each group), i.e. controls, only HIV, or HIV/hepatitis B virus (HBV) double infected, were studied, as concerned the following systemic immune parameters: immunoglobulin (IgG, IgM, IgA, IgD) levels; absolute numbers of blood CD+4, CD+8, CD+16 and CD+19 cells; phytohaemagglutinin (PHA)-blast responsiveness of T lymphocytes; natural killer (NK) cell activity--as tested by means of cytotoxicity assays; per cent suppression of PHA-dependent T cell blastogenesis in the presence of concanavalin A (Con A) selected T suppressor (Ts) cells. On the other hand, in 15 ARC-shifting cases belonging to HIV, and HIV/HBV groups, respectively, a second serum sample was collected and searched comparatively with the corresponding first serum sample, as regarded: presence of total and anti-p24 HIV antibodies, patterns of Western Blot (WB), as well as amounts of free p24-HIV antigen. In asymptomatic double HIV/HBV infected subjects, some immune disorders occurred, at a more significant degree, as compared to only HIV-infected. Once the shift toward ARC being installed, in both infected groups a decrease of anti-p24 HIV antibody presence, disappearance of corresponding band in WB confirmation test, as well as presence of free p24 antigen in serum, were noticed. However, greater amounts of p24 antigen in HIV/HBV infected, as compared to only HIV infected patients, were found. Some considerations about diagnostic and predictive value of presented data are discussed.

Some seroepidemiological data on HIV-antibody and HBs-antigen prevalence in children and infants of Craiova (Romania).

By testing 2362 children and infants from Craiova (Romania), for both HIV-antibody and HBs-antigen presence in the blood, a high rate of positivity was noticed in orphanages and in dystrophia units. A rate of "signal" (i.e. about 4% for HIV-antibodies, and 2.5% for HIV-antibody/HBs antigen), in hospitalized patients (pediatric service), was found as well. Four reasons support the horizontal way of virus transmission as the main route: most of seropositive subjects belong to 1-to-3-year age range, suggesting a virtual contamination before introduction of disposable syringes in Romania care units and hospitals; many HIV-seropositive cases have received frequent parenteral treatments, during repeated hospitalizations for acute respiratory disease; only few of the mothers of seropositive infants exhibited HIV-antibody presence; a relatively high rate of "double" seropositivity, i.e. HIV-antibody/HBs antigen, within tested serum samples was noticed.

HIV-antibody detection in children by competitive and direct micro-ELISA techniques.

A comparative study was carried out on 110 sera from children or infants, suspected of HIV-antibody presence following several micro-ELISA assays, using four direct micro-ELISA (Wellcozyme HIV 1 + 2, Rapid Elavia Mixt, Ortho Diagnostics, RECVIH) and a competitive system--Wellcozyme-Recombinant. In three of the four direct systems, as well as in the competitive system, significantly higher mean values of sample/cut off, and cut off/sample ratios, respectively, as compared to the direct systems RECVIH, were present. High optimal levels of sensitivity and specificity (%), as related to Western Blot results, were found with Wellcozyme direct and competitive kits, as well as with Rapid Elavia Mixt kit, as compared to lower levels exhibited by the other two direct system kits (Ortho Diagnostics an especially RECVIH). As regards three Western Blot undetermined results, obtained in patients with a severe clinical state and evolution to exitus, by comparing some serological markers of HIV infection in two serum samples belonging to the same case (second sample collected 4 weeks after collection of the first homologous sample), the disappearance of gag-encoded-p24 band in Western Blot, associated with negativation of HIV-p24-antibody and with the presence of free virus antigen in all three second serum samples occurred, that would reflect a probable fall of immune anti-HIV "barriers" during final stages of illness. Although Western Blot confirmation cannot be excluded, it seems to be useful to assay comparatively HIV-antibody presence by means of direct and competitive micro-ELISA systems, in the same serum sample.