Can Seminal IL-8 Level Be Used as a Marker of Leukocytospermia and Does It Have Any Correlation with Semen Parameters in Infertile Couples?

OBJECTIVES: Infection of male genital tract leads to leukocytospermia which may have a detrimental effect on semen quality. This study was conducted to evaluate whether seminal IL-8 level can be used as a marker of leukocytospermia and does it have any correlation with semen parameters in infertile couples? METHODS: This cross-sectional study was conducted in an infertility clinic of a tertiary care hospital including 150 male partners of infertile couples who underwent semen analysis (WHO laboratory manual for the examination and processing of human semen, 5th edn, World Health Organization, Geneva, p 271, 2010), semen culture sensitivity and seminal IL-8 levels. Independent t-test, Mann-Whitney U test and Chi-square test were applied for analysis. RESULTS: Mean seminal plasma IL-8 level of patients with leukocytospermia was significantly higher than patients without leukocytospermia (1143.67 ± 887.03 vs. 267.174 ± 242.29, p value < 0.001). Strong positive correlation was found between seminal plasma IL-8 levels and pus cells in the semen (r = 0.950, p < 0.001); AUC for seminal plasma IL-8 was 0.985 (CI 0.972-0.988), and a cutoff value of 399 pg/ml was determined to diagnose leukocytospermia. This value had high sensitivity (91.8%), specificity (94.5%), positive predictive value (94.4%) and diagnostic accuracy (93.2%) for detecting leukocytospermia. Seminal IL-8 levels correlated negatively with sperm motility (r = - 0.29, p < 0.001) and morphology (r = - 0.230, p < 0.01). CONCLUSION: Seminal plasma IL-8 levels were found to be almost five times higher in male partners with leukocytospermia than in non-leukocytospermia group, and it appears to be a promising tool to detect leukocytospermia. Seminal IL-8 level correlated negatively with semen parameters including sperm motility and morphology.

Distribution, antimicrobial resistance and predictors of mortality in neonatal sepsis.

OBJECTIVE: The aim of this study is to investigate etiological agents, patterns of antimicrobial resistance and predictors of mortality in culture proven neonatal sepsis. METHOD: This is a twenty-four month retrospective cohort study of infants with culture proven sepsis. Demographic data, type of isolates and its sensitivity pattern were recorded. Multidrug resistant gram-negative isolates were defined as resistance to any three of five antibiotic classes: extended-spectrum cephalosporins, carbapenems, aminoglycosides, fluoroquinolones and piperacillin-tazobactam. RESULT: A total of 183 case with culture positive sepsis were identified. Early onset sepsis occurred in 59% of cases. The majority of isolates (56.2%) were gram-positive but the most common individual isolates were klebsiella spp. (31.1%), Staphylococcus aureus (24.5%) and coagulase-negative staphylococci (CONS) (22.9%). The pathogen mix in early-onset did not differ from late-onset sepsis. High rates of multidrug resistance were observed in klebsiella spp. (49.1%), Escherichia coli (50%), citrobacter spp (50%), acinetobacter spp. (28.5%), pseudomonas spp. (100%) isolates. Methicillin resistance prevailed in 16.6% of coagulase-negative staphylococci, 24.4% of Staphylococcus aureus and 62.5% of enterococcus spp. Multivariate analysis revealed invasive ventilation and early onset sepsis to be independently associated with increased risk of mortality in contrast to breast milk feeding which is associated with decreased risk of mortality. CONCLUSION: A high degree of antimicrobial resistance underscores the need to understand the pathogenesis of resistance, curtail the irrational prescription of antibiotics in neonates and the requirement for measures to prevent it in low-income and middle-income countries.

Role of prophylactic antibiotics in neonates born through meconium-stained amniotic fluid (MSAF)--a randomized controlled trial.

UNLABELLED: The objective of the study was to evaluate the effect of administering prophylactic antibiotics on the development of neonatal sepsis in term neonates born through meconium-stained amniotic fluid (MSAF). Two hundred and fifty eligible neonates were randomized to study group (Antibiotic group-receiving first-line antibiotics for 3 days) and control group (No Antibiotic group). Both groups were evaluated clinically and by laboratory parameters (sepsis screen and blood cultures) for development of sepsis. All neonates were monitored for respiratory, neurological, and other systemic complications and received supportive treatment according to standard management protocol of the unit. One hundred and twenty one neonates were randomized to 'Antibiotic' group and 129 to 'No Antibiotic' group. The overall incidence of suspect sepsis was 9.6 % in the study population with no significant difference between 'No Antibiotic' and 'Antibiotic' groups (10.8 vs. 8.2 %, p = 0.48, odds ratio (OR) 0.74, 95 % confidence interval (CI) 0.32-1.73). Incidence of culture-proven sepsis was also not significantly different between the two groups (5.42 vs. 4.13 %, p = 0.63, OR 0.75, 95 % CI 0.23-2.43). The incidence of mortality, meconium aspiration syndrome, and other complications was comparable amongst the two groups. CONCLUSION: Routine antibiotic prophylaxis in neonates born through MSAF did not reduce the incidence of sepsis in this study population. (Clinicaltrials.gov no. - NCT01290003).

Incidence of bla NDM-1 gene in Escherichia coli isolates at a tertiary care referral hospital in Northeast India.

PURPOSE: Increasing reports on New Delhi metallo-ß-lactamase-1 (NDM-1) producing Escherichia coli constitute a serious threat to global health since it is found to be highly resistant to most of the currently available antibiotics including carbapenems. This study has been performed to find out the incidence blaNDM-1 in E. coli isolates recovered from the various clinical samples at a tertiary care referral hospital in Northeast India. MATERIALS AND METHODS: A total of 270 non-duplicated E. coli isolates were recovered from the various clinical samples at a tertiary care referral hospital in Northeast India. All isolates with reduced susceptibility to meropenem or ertapenem (diameter of zones of inhibition, ≤ 21 mm) were further phenotypically confirmed for carbapenemase production by modified Hodge test. All screened isolates were also subjected to the polymerase chain reaction detection of blaNDM-1 gene and additional bla genes coding for transmission electron microscopy, SHV, CTX-M, and AmpC. RESULTS: Out of 270 E. coli isolates, 14 were screened for carbapenemase production on the basis of their reduced susceptibility to meropenem or ertapenem. All screened isolates were found to be positive for blaNDM-1 . Each of the blaNDM-1 possessing isolate was also positive for two or more additional bla genes, such as blaTEM , blaCTX-M and blaAmpC . Phylogenetic analysis showed very less variation in blaNDM-1 gene with respect to blaNDM-1 possessing E. coli isolates from other parts of India and abroad. CONCLUSIONS: Our findings highlight the incidence of blaNDM-1 in E. coli isolates with a reduced susceptibility to meropenem or ertapenem.

Association of bacterial vaginosis with adverse fetomaternal outcome in women with spontaneous preterm labor: a prospective cohort study.

OBJECTIVE: To compare the fetomaternal outcome in women with spontaneous preterm labor, with or without bacterial vaginosis (BV). METHODS: One hundred and fifty-two pregnant patients presenting with spontaneous preterm labor between 28 and 35 weeks of gestation were screened for BV using Amsel's criteria and Nugent score, and were divided into two groups of 30 patients each, based on the BV positive or negative screen. Both the groups were followed till puerperium, and the fetal-maternal outcome was studied. The data was analyzed using Chi-square test and Man-Whitney test. RESULTS: BV was detected in 37 out of 152 women with preterm labor (24.34%). There was a significant increase in the incidence of respiratory distress (14% vs. 6%), requirement of intermittent positive pressure ventilation (IPPV) (14% vs. 5%), admission in neonatal intensive care unit (NICU) (15% vs. 6%), and duration of NICU stay >2 days (15% vs. 6%) in patients with BV. No significant difference was found in the mean birth weight, Apgar score, incidence of neonatal sepsis, perinatal mortality, and postpartum fever between the two groups. CONCLUSIONS: BV is a risk factor for increased neonatal morbidity. More research is needed for designing appropriate screening and treatment guidelines for prevention of adverse outcomes associated with BV.

Paratyphoid sepsis.

A large for gestational age male baby was born to a healthy young primigravida, on L-thyroxime, at 40 weeks by caesarean delivery in a tertiary care hospital. The baby had episodes of hypoglycemia during his immediate four postnatal days in the nursery that were successfully managed with intravenous glucose administration. The baby became unwell on day 5 and had a positive sepsis-screening test. Blood culture revealed a multidrug susceptible S. Paratyphi A strain, which he probably acquired on the first or second postnatal day from the contaminated expressed breast milk or the formula feed.

Rare serotype non-typhoidal Salmonella sepsis.

An 11-month female with a poor socio-economic status presented to a tertiary care paediatric hospital with complaints of fever of 4-5 days and diarrhoea of 20 days duration. The patient didn't respond to the prescribed antimicrobials namely--Norfloxacin and metronidazole. On admission she was diagnosed as persistent diarrhea with PEM grade III with sepsis. Stool examination and culture were negative for any pathogens, however blood culture yielded growth of Salmonella Virchow which was susceptible to most common antimicrobial agents excepting Trimethoprim Sulfamethoxazole. Salmonella Virchow is a common non-typhoidal Salmonellae causing bacteremia in the west, however this is the first report of bacteremia by S. virchow from India.

Antimicrobial resistance of enterococcal blood isolates at a pediatric care hospital in India.

Enterococci are one of the leading causes of nosocomial infections. In recent years, enterococci have become increasingly resistant to a wide range of antimicrobial agents. From April to October 2001, a study was conducted to speciate and determine the antimicrobial susceptibility of 50 isolates of enterococci from bacteremic children. These isolates were tested for antimicrobial susceptibility to the commonly used antibiotics. Screening for vancomycin resistance was done by the agar screen method, and the results were confirmed by determining the minimum inhibitory concentration (MIC) using the agar dilution method. It was observed that 33 isolates were Enterococcus faecium, followed by E. faecalis (10), E. durans (4), and E. dispar (3). Seventy-two percent of strains were resistant to ampicillin, 46% to amoxicillin + clavulanic acid, 72% to ciprofloxacin, 54% to doxycyclin, and 74% to erythromycin. Sixty-six percent of isolates showed high-level gentamicin resistance and 42% showed high-level streptomycin resistance. Four strains showed raised MIC to vancomycin (8 microg/ml). It was concluded that multidrug resistant E. faecium is emerging as an important agent of bacteremia in children.

Microbiological profile of neonatal septicemia in a pediatric care hospital in Delhi.

Septicemia continues to be an important cause of morbidity and mortality in the neonatal units and periodic review of cases to assess any changing trends in the infecting organisms and their antimicrobial susceptibility is important. Over a period of one year (July 2000 to June 2001), 632 samples of blood cultures were submitted to the bacteriology laboratory Microbiology, Lady Hardinge Medical College. These samples were investigated for microbial etiology and the isolates obtained were tested for their susceptibility to the commonly used antibiotics. Twenty per cent (125) cases were culture positive. Gram-negative bacteria were the predominant isolates (62%), commonest being Klebsiella pneumoniae (34%) followed by E. coli (17%), Acinetobacter spp. (9%) and Enterobacter aerogenes (2%). Gram-positive cocci were isolated in 20% cases, of which coagulase negative staphylococcus was the predominant isolate (11%) followed by Enterococcus spp. (5%) and S. aureus (4%). Candida spp. was isolated from 18% of cases. Resistance to commonly used antibiotics was seen in more than 35% of isolates. An alarming observation was the very high incidence of resistance to amoxycillin+clavulanic acid and ceftriaxone (>80%). All isolates showed highest susceptibility to ciprofloxacin.

Aminoglycoside resistance in enterococci isolated from paediatric septicaemia in a tertiary care hospital in north India.

BACKGROUND & OBJECTIVES: Enterococci are important nosocomial agents and serious infections caused by them are often treated with a combination of cell wall inhibitor and aminoglycoside. However, the presence of high level aminoglycoside resistance in these isolates makes this treatment combination ineffective. The prevalence of such isolates in a tertiary care set up has important diagnostic and therapeutic implications. The present study was carried out to find out the occurrence of high level aminoglycoside resistant isolates of enterococci in paediatric septicaemia cases in a tertiary care set up in north India. METHODS: Blood of paediatric cases with a clinical diagnosis of septicaemia was cultured to isolate and identify enterococci. Agar screen method was used to detect high level streptomycin and gentamicin resistance in these isolates. Vancomycin susceptibility of these isolates were determined as per the National Committee for Clinical Laboratory Standards (NCCLS) recommendations. RESULTS: Fifty one enterococcal strains were isolated from 21 neonates, nine infants and 21 children with a clinical diagnosis of septicaemia. Sixty eight per cent of these isolates had high level gentamicin resistance and forty three per cent had high level streptomycin resistance. All the isolates with high level streptomycin resistance also had high level gentamicin resistance. More than ninety five per cent of these isolates were sensitive to vancomycin. INTERPRETATION & CONCLUSION: The occurrence of high level gentamicin and high level streptomycin resistance in enterococcal isolates in our set up was high. This would require routine testing of the enterococcal isolates for high level aminoglycoside resistance. Alternative treatment regimes need to be sought, besides prudent use of antibiotics.

An outbreak of candidemia due to Candida tropicalis in a neonatal intensive care unit.

An outbreak of candidemia due to Candida tropicalis involving 16 neonates (gestational age 28-36 weeks) is reported. All infants had received hyperalimentation and at least one course of antibiotics. The commonest clinical manifestations included episodes of acute respiratory distress and lack of response to antibacterial antibiotic therapy. Candida tropicalis was recovered from blood in all the 16 infants and urine cultures were positive in 14 infants. Environmental sampling yielded C. tropicalis from one each of the blankets and mattresses used for neonates. Four of five urinary tract isolates and both environmental isolates genotyped by arbitrarily primed-PCR with several random primers were shown to belong to the same genotype.

Changing trends of Candida species in neonatal septicaemia in a tertiary North Indian hospital.

Four hundred and fifty four blood samples of clinically diagnosed septicemic neonates were collected over a period of six months from the neonatal ICU of Kalawati Saran Children Hospital, New Delhi. 144 samples were culture positive; out of which 50 (34.7%) were Candida isolates. 92% isolates were Candida tropicalis, 4% were C. albicans and C. kefyr each. The study emphasises the changing pattern of Candida species and their importance in blood stream infections in neonates.

Differential effects of phosphatidylinositol 3-kinase inhibition on intracellular signals regulating GLUT4 translocation and glucose transport.

Phosphatidylinositol (PI) 3-kinase is required for insulin-stimulated translocation of GLUT4 to the surface of muscle and fat cells. Recent evidence suggests that the full stimulation of glucose uptake by insulin also requires activation of GLUT4, possibly via a p38 mitogen-activated protein kinase (p38 MAPK)-dependent pathway. Here we used L6 myotubes expressing Myc-tagged GLUT4 to examine at what level the signals regulating GLUT4 translocation and activation bifurcate. We compared the sensitivity of each process, as well as of signals leading to GLUT4 translocation (Akt and atypical protein kinase C) to PI 3-kinase inhibition. Wortmannin inhibited insulin-stimulated glucose uptake with an IC(50) of 3 nm. In contrast, GLUT4myc appearance at the cell surface was less sensitive to inhibition (IC(50) = 43 nm). This dissociation between insulin-stimulated glucose uptake and GLUT4myc translocation was not observed with LY294002 (IC(50) = 8 and 10 microm, respectively). The sensitivity of insulin-stimulated activation of PKC zeta/lambda, Akt1, Akt2, and Akt3 to wortmannin (IC(50) = 24, 30, 35, and 60 nm, respectively) correlated closely with inhibition of GLUT4 translocation. In contrast, insulin-dependent p38 MAPK phosphorylation was efficiently reduced in cells pretreated with wortmannin, with an IC(50) of 7 nm. Insulin-dependent p38 alpha and p38 beta MAPK activities were also markedly reduced by wortmannin (IC(50) = 6 and 2 nm, respectively). LY294002 or transient expression of a dominant inhibitory PI 3-kinase construct (Delta p85), however, did not affect p38 MAPK phosphorylation. These results uncover a striking correlation between PI 3-kinase, Akt, PKC zeta/lambda, and GLUT4 translocation on one hand and their segregation from glucose uptake and p38 MAPK activation on the other, based on their wortmannin sensitivity. We propose that a distinct, high affinity target of wortmannin, other than PI 3-kinase, may be necessary for activation of p38 MAPK and GLUT4 in response to insulin.

Insulin accelerates inter-endosomal GLUT4 traffic via phosphatidylinositol 3-kinase and protein kinase B.

Insulin enhances plasmalemmal-directed traffic of glucose transporter-4 (GLUT4), but it is unknown whether insulin regulates GLUT4 traffic through endosomal compartments. In L6 myoblasts expressing Myc-tagged GLUT4, insulin markedly stimulated the rate of GLUT4myc recycling. In myoblasts stimulated with insulin to maximize surface GLUT4myc levels, we followed the rates of surface-labeled GLUT4myc endocytosis and chased its intracellular distribution in space and time using confocal immunofluorescence microscopy. Surface-labeled GLUT4myc internalized rapidly (t(12) 3 min), reaching the early endosome by 2 min and the transferrin receptor-rich, perinuclear recycling endosome by 20 min. Upon re-addition of insulin, the t(12) of GLUT4 disappearance from the plasma membrane was unchanged (3 min), but strikingly, GLUT4myc reached the recycling endosome by 10 and left by 20 min. This effect of insulin was blocked by the phosphatidylinositol 3-kinase inhibitor LY294002 or by transiently transfected dominant-negative phosphatidylinositol 3-kinase and protein kinase B mutants. In contrast, insulin did not alter the rate of arrival of rhodamine-labeled transferrin at the recycling endosome. These results reveal a heretofore unknown effect of insulin to accelerate inter-endosomal travel rates of GLUT4 and identify the recycling endosome as an obligatory stage in insulin-dependent GLUT4 recycling.

Hyperosmolarity reduces GLUT4 endocytosis and increases its exocytosis from a VAMP2-independent pool in l6 muscle cells.

The intracellular traffic of the glucose transporter 4 (GLUT4) in muscle cells remains largely unexplored. Here we make use of L6 myoblasts stably expressing GLUT4 with an exofacially directed Myc-tag (GLUT4myc) to determine the exocytic and endocytic rates of the transporter. Insulin caused a rapid (t(12) = 4 min) gain, whereas hyperosmolarity (0.45 m sucrose) caused a slow (t(12) = 20 min) gain in surface GLUT4myc molecules. With prior insulin stimulation followed by addition of hypertonic sucrose, the increase in surface GLUT4myc was partly additive. Unlike the effect of insulin, the GLUT4myc gain caused by hyperosmolarity was insensitive to wortmannin or to tetanus toxin cleavage of VAMP2 and VAMP3. Disappearance of GLUT4myc from the cell surface was rapid (t(12) = 1.5 min). Insulin had no effect on the initial rate of GLUT4myc internalization. In contrast, hyperosmolarity almost completely abolished GLUT4myc internalization. Surface GLUT4myc accumulation in response to hyperosmolarity was only partially blocked by inhibition of tyrosine kinases with erbstatin analog (erbstatin A) and genistein. However, neither inhibitor interfered with the ability of hyperosmolarity to block GLUT4myc internalization. We propose that hyperosmolarity increases surface GLUT4myc by preventing GLUT4 endocytosis and stimulating its exocytosis via a pathway independent of phosphatidylinositol 3-kinase activity and of VAMP2 or VAMP3. A tetanus toxin-insensitive v-SNARE such as TI-VAMP detected in these cells, might mediate membrane fusion of the hyperosmolarity-sensitive pool.