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1.
Isr Med Assoc J ; 21(2): 71-76, 2019 Feb.
Article in English | MEDLINE | ID: mdl-30772954

ABSTRACT

BACKGROUND: Endothelial progenitor cells may have a role in ongoing endothelial repair. Impaired mobilization or depletion of these cells may contribute to progression of vascular disease. Our hypothesis was that endothelial progenitor cells would be suppressed in patients with acute cerebrovascular event based on our previous study that found severe endothelial dysfunction in those patients. OBJECTIVES: To study the ability of patients with acute stroke to build colonies of endothelial progenitor cells. METHODS: We studied the number of colony-forming units of endothelial progenitor cells (CFU-EPCs) from the peripheral blood of 22 male patients with a first-time acute stroke (age 58.09 ± 9.8 years) and 13 healthy men (34 ± 6.7 years), 8 female patients with a first-time acute stroke (54.6 ± 10.3 years) and 6 healthy women (38.3 ± 11.6 years). Endothelium-dependent function was assessed by high-resolution ultrasonography of the brachial artery that measured the change in diameter of the artery by flow-mediated diameter percent change (FMD%). All patients had strokes demonstrated by a brain computed tomography (CT) scan done on admission. Peripheral blood was drawn soon after admission and was processed for endothelial progenitor cells in culture. RESULTS: Thirty patients without known cardiovascular risk factors and who did not take any medications were admitted with a first-time acute stroke. All demonstrated a strong correlation between CFU-EPCs grown in culture and endothelial dysfunction (r = 0.827, P < 0.01). Endothelial dysfunction with an FMD% of -2.2 ± 9.7% was noted in male patients vs. 17.5 ± 6.8% in healthy males (P = 0.0001), and -7.2 ± 10.1% in female patients vs. 25.1 ± 7.1% in healthy females (P = 0.0001). CFU-EPCs were 5.5 ± 6.3 in men with stroke vs. 23.75 ± 5.3 in healthy males (P = 0.0001), and 7.6 ± 4.9 in women with stroke vs. 22.25 ± 6.7 in healthy females (P = 0.0004).


Subject(s)
Brain Ischemia/blood , Endothelial Progenitor Cells/metabolism , Endothelium, Vascular/metabolism , Stroke/blood , Adult , Brain Ischemia/complications , Female , Humans , Male , Middle Aged , Prospective Studies , Stroke/complications
2.
BMC Infect Dis ; 19(1): 72, 2019 Jan 18.
Article in English | MEDLINE | ID: mdl-30658585

ABSTRACT

BACKGROUND: Rapid and accurate pathogen identification in blood cultures is very important for septic patients and has major consequences on morbidity and mortality rates. In recent years, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS)-based technology has become useful for highly specific and sensitive identification of bacteria and yeasts from clinical samples including sterile body fluids. Additional in-house methods enabled direct identification from blood cultures following various preparation protocols. METHODS: Blood culture (5 ml) was harvested from each positive bottle following growth identification by BACTEC™ FX system and transferred into a VACUETTE® Z Serum Sep Clot Activator tube containing an inert gel, which following centrifugation separates microorganisms from the blood cells. We used MALDI-TOF MS analysis for identification of microorganisms collected from the gel surface. RESULTS: Positive blood culture bottles (186) were collected. In comparison with the routine method, 99% (184/186) and 90% (168/186) of the isolates were correctly identified by the SepsiTyper kit and the in-house method, respectively. We found high concordance (Pearson coefficient = 0.7, p <  0.0001) between our in-house method and the SepsiTyper kit. Additionally, high correlation was found in sub-groups of identified bacteria, with Pearson coefficients of 0.77 (p <  0.0001), 0.67 (p <  0.0001), and 0.73 (p <  0.007) for Gram negative, Gram positive, and anaerobic bacteria, respectively. CONCLUSIONS: Our in-house method was found to be in good agreement with the SepsiTyper kit. Considering the low costs and the rapid and easy implementation of this procedure, we propose our in-house method for the direct identification of bacteria from blood cultures.


Subject(s)
Bacteremia/microbiology , Fungemia/microbiology , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Bacteria/isolation & purification , Bacteriological Techniques/instrumentation , Bacteriological Techniques/methods , Blood Culture , Fungi/isolation & purification , Humans
3.
Clin Exp Med ; 19(2): 255-260, 2019 May.
Article in English | MEDLINE | ID: mdl-30673918

ABSTRACT

Diabetic retinopathy is the most severe ocular complication of diabetes and may lead to visual disability and blindness. Proliferative diabetic retinopathy (PDR) is characterized by ischemia-induced neovascularization with associated complications. An association was established between the presence of PDR, cardiovascular disease, and mortality among patients with type 1 diabetes mellitus and type 2 diabetes mellitus in epidemiological studies. However, the mechanism underlying increased cardiovascular risk in patients with PDR is still unknown. In recent years, a group of miRNAs has been linked to the pathology of diabetes mellitus. Besides, miRNAs in biofluids such as serum have been suggested as potential minimally invasive biomarkers of diabetes and vascular complications. This was a prospective study that recruited 40 human subjects: 10 healthy subjects, 10 with diabetes but without retinopathy (NDR), 10 with diabetic non-proliferative retinopathy (NPDR), and 10 with proliferative diabetic retinopathy (PDR). To examine whether serum miRNAs show altered levels at different stages of diabetic retinopathy, seven specific miRNA candidates (miR-126-3p, miR-130a-3p, miR-21-1, let-7f-5p, miR-122, miR-30c and miR-451a) were measured by qRT-PCR in RNA isolated from sera of all subjects. miR-122 levels increased in parallel with retinopathy severity: from healthy controls to NDR and from NDR to NPDR. However, when the disease progressed to PDR a marked decrease in miR-122 level was noted. This decrease was significant both compared to NPDR samples (p = 0.016) and to all non-PDR samples (p = 0.0002). Additionally, a positive trend was observed comparing miR-122 levels and the number of endothelial progenitor cells in the sera of all subjects. A significant increase in miR-122 was found in patients with diabetic retinopathy that may be related to its role in preventing angiogenesis and proliferation. The dramatic decline in patients with PDR may represent an inhibition or exhaustion of the anti-angiogenic anti-proliferative defense system. Further studies are needed to understand whether miRNA-122 has a role in the pathogenesis of diabetic retinopathy.


Subject(s)
Diabetic Retinopathy/pathology , MicroRNAs/blood , Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Prospective Studies , Real-Time Polymerase Chain Reaction , Serum/chemistry
4.
Int J Med Mushrooms ; 20(1): 71-80, 2018.
Article in English | MEDLINE | ID: mdl-29604914

ABSTRACT

We studied the anti-Leishmania activity of a fractionated extract from the mushroom Morchella importuna in an in vitro system. Leishmaniasis is an important infectious disease caused by a range of Leishmania species, which are multihost protozoa parasites transmitted to humans by the sand fly and infecting macrophages. Leishmaniasis is an increasing worldwide health problem, including in the Mediterranean basin. Current chemotherapy treatments are limited by their toxic effects, the need for long-term treatment, and the increasing development of resistance by the parasite cells. Thus, alternative therapies are being considered, including herbal and mushroom products. We studied the effect of extracts from M. importuna on L. tropica promastigote cell proliferation and survival, and on their toxicity against human macrophages. The aqueous mushroom extract was compared with 3 successive extracted fractions: an 80% ethanol fraction, a water-soluble polysaccharide fraction, and a polyphenolic fraction. All 4 extracts showed anti-Leishmania activity; the aqueous extract was most active. The inhibition activity was dose dependent in killing Leishmania. No cell recovery was recorded after exposure to the mushroom extract. Microscopic observation showed morphological changes and the loss of flagella on the parasites. No cytotoxic activity was recorded against human macrophages at the same extract concentrations. The findings suggest the potential use of extracts of an edible Morchella mushroom against the Leishmania parasite in humans.


Subject(s)
Antiprotozoal Agents/pharmacology , Ascomycota/chemistry , Leishmania/drug effects , Antiprotozoal Agents/isolation & purification , Drug Discovery , Flagella/drug effects , Humans , L-Lactate Dehydrogenase/metabolism , Leishmania/ultrastructure , Leishmaniasis/drug therapy , Leishmaniasis/parasitology , Macrophages/drug effects , Water
5.
J Glob Antimicrob Resist ; 12: 175-178, 2018 03.
Article in English | MEDLINE | ID: mdl-29033302

ABSTRACT

OBJECTIVES: Helicobacter pylori is a bacterial pathogen causing inflammation of the gastric mucosa that may lead to peptic ulcer, perforation or malignancy. Children are at risk of contracting H. pylori and developing subsequent morbidity. Diagnosis and management in children are difficult and merit a different approach compared with adults. This study aimed to describe the antimicrobial resistance rates of H. pylori to amoxicillin, tetracycline, clarithromycin, metronidazole, levofloxacin and rifampicin. METHODS: Biopsies (n=154) collected during endoscopic examinations were cultivated for 10days using a growth medium selective for H. pylori, of which 89 were H. pylori-positive. Antimicrobial resistance of the strains was assessed by Etest to establish minimum inhibitory concentrations (MICs) according to British Society for Antimicrobial Chemotherapy guidelines. RESULTS: Resistance rates were most notable for amoxicillin and clarithromycin at 12% and 35% with MICs of 0.74µg/mL and 2.51µg/mL, respectively. Resistance rates to tetracycline and levofloxacin were 8% and 2% with MICs of 2.57µg/mL and 2.0µg/mL, respectively. Resistance rates to rifampicin and metronidazole were 3% and 8% with MICs of 2.0µg/mL and 9.71µg/mL, respectively. CONCLUSION: Current rising antibiotic resistance rates for H. pylori are of concern. Performance of culture enables determination of the susceptibility profile, which may lead to a better choice of, and perhaps narrower spectrum, antibiotic agent. In light of these findings, we suggest that optimising the choice of antibiotic agent in children with H. pylori infection remains a challenge for clinicians and thus requires further investigation in randomised clinical trials.


Subject(s)
Anti-Bacterial Agents/pharmacology , Helicobacter Infections/microbiology , Helicobacter pylori/drug effects , Adolescent , Amoxicillin/pharmacology , Biopsy , Child , Child, Preschool , Clarithromycin/pharmacology , Drug Resistance, Multiple, Bacterial , Female , Helicobacter pylori/genetics , Helicobacter pylori/isolation & purification , Humans , Israel , Levofloxacin/pharmacology , Male , Metronidazole/pharmacology , Microbial Sensitivity Tests , Tetracycline/pharmacology
6.
Cytokine ; 106: 76-79, 2018 06.
Article in English | MEDLINE | ID: mdl-29133026

ABSTRACT

BACKGROUND: Proliferative diabetic retinopathy is a devastating complication of diabetes mellitus, developing within 15 years in 50% of patients with type 1 diabetes mellitus (DM) and in 10% of patients with type 2 DM. The correlation between levels of inflammatory markers in the peripheral blood and retinopathy staging has not been studied yet, and the purpose of this prospective study was to find a possible association between inflammation and staging of diabetic retinopathy. METHODS: A prospective (pilot) study that measured level of adhesion molecules in the peripheral blood of 10 healthy subjects and 30 patients with type 2 diabetes mellitus. Patients were grouped by the degree of retinopathy: 10 without retinopathy, 10 with non-proliferative retinopathy [NPDR] and 10 with proliferative retinopathy [PDR]. After signing the consent form, an ophthalmologic examination was performed, and 10 mL of blood was drawn. In order to assess adhesion molecules' level serum samples were collected, frozen, and stored at a temperature of -80 °C until analysis was performed as one batch. RESULTS: 10 healthy volunteers and 30 patients were enrolled. Healthy volunteers were younger (36.6 ±â€¯7.9 years) compared to patients (no retinopathy 64.5 ±â€¯10.8 years, NPDR 71.4 ±â€¯8.9 years, and PDR 63.3 ±â€¯11.6 years) (p = .0003 for all groups of patients in comparison with the healthy subjects). VCAM-1 levels were increased by retinopathy staging - starting from 81.86 ±â€¯3.80 ng/ml (healthy), 105.55 ±â€¯1.37 ng/ml (no retinopathy), 111.78 ±â€¯4.14 ng/ml (NPDR), and 123.45 ±â€¯3.99 ng/ml (PDR), with a significant difference between healthy and patients without retinopathy (p = .03), between no retinopathy and NPDR (p = .001), and between NPDR and PDR (p < .0001). E selectin was increased in correlation with severity of the retinopathy, with a significant difference between groups of patients (p = .03 between healthy subjects and T2DM patients without retinopathy, p = .001 between patients with T2DM no retinopathy and NPDR, p < .0001 between NPDR and PDR). CONCLUSIONS: We found a significant increase in levels of adhesion molecules (VCAM-1) and selectins (E-selectin) in parallel with increased severity of diabetic retinopathy, with a significant difference of inflammatory markers between stages of retinopathy.


Subject(s)
Diabetic Angiopathies/blood , Diabetic Angiopathies/complications , Diabetic Retinopathy/blood , Diabetic Retinopathy/complications , E-Selectin/blood , Microvessels/pathology , Vascular Cell Adhesion Molecule-1/blood , Adult , Aged , Biomarkers/blood , Female , Humans , Male , Middle Aged , Selectins/blood
7.
Helicobacter ; 22(6)2017 Dec.
Article in English | MEDLINE | ID: mdl-29058343

ABSTRACT

BACKGROUND: There are several methods for Helicobacter pylori infection diagnosis. AIM: The efficacies of three methods for H. pylori identification directly from a biopsy were compared: histology, culture, and molecular GenoType® HelicoDR test. MATERIALS & METHODS: Eighty-five triplicates of stomach antrum biopsies were obtained during gastroscopy procedures for culture, histology, and molecular assay. In addition, we performed molecular identification of genes encoding resistance to clarithromycin and fluoroquinolones. RESULTS: The results have shown that the most specific method with the highest number of positive specimens was by molecular kit, compared to culture and histology (94.3%, 77.1%, and 71.4%, respectively). There was a higher rate of resistance mutations to clarithromycin than to fluoroquinolones (68.26% vs 20%). The most common mutations for clarithromycin and fluoroquinolones resistance were found in alleles A2143G and N87K, respectively. The highest rate of positive specimens was identified by the molecular. DISCUSSION: GenoType HelicoDR kit (94.3%), which has several advantages: direct identification, strain resistance characterization, mixture of genotypes detection, and no transport or storage limitations; thus, it is an excellent epidemiological screening tool. This work has demonstrated a lower resistance rate to fluoroquinolones; it is possible that in the investigated geographic area treatment with fluoroquinolones may be preferable to clarithromycin. GenoType® HelicoDR test eliminates the need for culture performance and susceptibility tests for several common antibiotic agents and enables optimal and specific antibiotic treatment adjustment. CONCLUSION: We recommend a combination of PCR assay and bacterial culture for a quick method of screening and more efficient identification of H. pylori strains and resistance patterns.


Subject(s)
Anti-Bacterial Agents/pharmacology , Bacteriological Techniques/methods , Drug Resistance, Bacterial , Helicobacter Infections/diagnosis , Helicobacter pylori/isolation & purification , Histocytochemistry/methods , Molecular Diagnostic Techniques/methods , Adult , Child , Clarithromycin/pharmacology , Female , Fluoroquinolones/pharmacology , Gastric Mucosa/microbiology , Gastric Mucosa/pathology , Helicobacter Infections/microbiology , Helicobacter Infections/pathology , Helicobacter pylori/drug effects , Helicobacter pylori/genetics , Humans , Male , Mass Screening/methods
8.
Harefuah ; 156(10): 631-634, 2017 Oct.
Article in Hebrew | MEDLINE | ID: mdl-29072380

ABSTRACT

INTRODUCTION: Among all infectious agents that cause gastrointestinal infection in children, the most common is the Campylobacter bacterium. The bacterium has multiple virulence factors such as motility, adhesion and invasion of the human intestinal lining, and enzyme secretion. In recent years, there has been a worldwide increase in Campylobacter resistance to antibiotics. AIMS: To examine the frequency of Campylobacter among children who were hospitalized at the Poriya Medical Center during 2012-2014 and suffered from an intestinal infection caused by Campylobacter; to compare the demographic, clinical, and laboratory data of Jewish and Arab children; to examine the resistance rate of the bacterium to antibiotics. METHODS: The data on Campylobacter frequency in children who suffered from an intestinal infection was extracted from the medical records: age, sex, hospitalization duration, hemoglobin and leukocyte values in blood chemistry, the residential environment, and antibiotic treatment during hospitalization. In addition, antibiotic susceptibility tests were performed for Erythromycin and Ciprofloxacin for all Campylobacter cultures that were isolated from patients' stool samples and kept frozen. RESULTS: Campylobacter is the most prevalent bacterial factor among children who were hospitalized following enteritis. There are differences in the bacterium frequency among Jewish children in comparison to frequency in Arab children in the following aspects: Campylobacter is more frequent in Arab children, more common among children living in rural areas, and especially those of Arab origin. Arab children were hospitalized for longer durations than Jewish children. The mean age of Jewish children who suffered from infection caused by Campylobacter was higher compared to the mean age of Arab children. No difference was found in leukocyte values in the cell count. Hemoglobin values were lower among Jewish children compared to Arab children. There was a high percentage of children treated with antibiotics due to intestinal infection caused by Campylobacter, especially among Arab children. Resistance to Erythromycin was not found; however the rate of resistance to Ciprofloxacin was 10.7%. CONCLUSIONS: There are significant differences in intestinal infection caused by Campylobacter among Jewish and Arab children in parameters such as: mean age, hospitalization duration, and residential area. The antibiotic resistance rate that was found is low; however, presently, it still exists.


Subject(s)
Anti-Bacterial Agents/therapeutic use , Campylobacter Infections/ethnology , Campylobacter/drug effects , Drug Resistance, Bacterial , Arabs , Campylobacter Infections/drug therapy , Campylobacter Infections/epidemiology , Child , Humans , Israel/ethnology , Judaism , Microbial Sensitivity Tests , Prevalence
9.
Harefuah ; 156(10): 642-644, 2017 Oct.
Article in Hebrew | MEDLINE | ID: mdl-29072383

ABSTRACT

INTRODUCTION: Helicobacter Pylori (H. Pylori) is a spiral shaped Gram-negative bacterium which is known to cause chronic gastric inflammation (gastritis) that could develop into a gastric or duodenal ulcer. The standard first line therapy for H. Pylori infection is a 7-14 days period of "triple therapy" consisting of proton pump inhibitors (PPI) and the antibiotics clarithromycin and amoxicillin or metronidazole. Recently there has been an increase in H. Pylori resistance to antibiotic treatment. Throughout the years 1999, 2002, 2010, 2013 and 2014 studies have been conducted in Israel that examined H. Pylori resistance rates for commonly used antibiotics. These studies included 40-138 participants who were diagnosed with infection caused by H. Pylori. Based on information derived from these studies, there is a clear increase in H. Pylori resistance to antibiotics, particularly to tetracycline, amoxicillin and clarithromycin.


Subject(s)
Anti-Bacterial Agents/therapeutic use , Drug Resistance, Bacterial , Helicobacter Infections/drug therapy , Helicobacter pylori/drug effects , Clarithromycin , Colony Count, Microbial , Drug Therapy, Combination , Humans , Israel , Microbial Sensitivity Tests
10.
J Glob Antimicrob Resist ; 10: 161-164, 2017 09.
Article in English | MEDLINE | ID: mdl-28729205

ABSTRACT

OBJECTIVES: An increase of Clostridium difficile isolates with reduced susceptibility to various antimicrobial agents has been observed, including isolates that are non-susceptible to antibiotics that are routinely used for treatment of C. difficile, such as vancomycin and metronidazole. We determined the susceptibility rates of C. difficile isolates from hospitals in northern Israel to various antibiotics including tigecycline, which was not previously reported from Israel. METHODS: A total of 81 stool samples were collected from three hospitals in northern Israel from patients with C. difficile infection. Specimens were screened for BI/NAP1/027 ribotype, cultured, and sensitivity tests were performed for vancomycin, metronidazole, moxifloxacin, and tigecycline. Statistical tests were applied for analysing the differences in distribution of resistance between the different antibiotics and between BI/NAP1/027 and resistance of antibiotics. RESULTS: Reduced susceptibility was found among 6/81 isolates for vancomycin, 4/81 for metronidazole, and 17/81 for moxifloxacin. Only 1 isolate had reduced susceptibility to tigecycline, with a mean MIC of 0.05µg/mL. Reduced susceptibility to moxifloxacin was significantly associated with reduced susceptibility to vancomycin (p=0.016) and to metronidazole (p=0.0276), and reduced susceptibility to metronidazole was associated with reduced susceptibility to vancomycin (p=0.0259). Eight of 81 isolates (9.9%) were positive for BI/NAP1/027 ribotype and had significantly higher non-susceptibility rates to moxifloxacin and vancomycin compared with BI/NAP1/027 negative isolates (p<0.0001 and p=0.0113, respectively). CONCLUSIONS: We found higher non-susceptibility rates to vancomycin and metronidazole than most previous studies, while tigecycline resistance rates are very low in northern Israel, rendering it a potential agent for treating CDI.


Subject(s)
Anti-Infective Agents/pharmacology , Clostridioides difficile/drug effects , Clostridioides difficile/isolation & purification , Clostridium Infections/microbiology , Microbial Sensitivity Tests , Anti-Bacterial Agents/pharmacology , Clostridioides difficile/pathogenicity , Drug Combinations , Feces/microbiology , Hospitals , Humans , Israel , Metronidazole/pharmacology , Ribotyping , Tigecycline/pharmacology , Vancomycin/pharmacology
11.
Sci Rep ; 7: 40114, 2017 01 06.
Article in English | MEDLINE | ID: mdl-28059171

ABSTRACT

Bacteria of the genus Legionella cause water-based infections resulting in severe pneumonia. Here we analyze and compare the bacterial microbiome of sputum samples from pneumonia patients in relation to the presence and abundance of the genus Legionella. The prevalence of Legionella species was determined by culture, PCR, and Next Generation Sequencing (NGS). Nine sputum samples out of the 133 analyzed were PCR-positive using Legionella genus-specific primers. Only one sample was positive by culture. Illumina MiSeq 16S rRNA gene sequencing analyses of Legionella-positive and Legionella-negative sputum samples, confirmed that indeed, Legionella was present in the PCR-positive sputum samples. This approach allowed the identification of the sputum microbiome at the genus level, and for Legionella genus at the species and sub-species level. 42% of the sputum samples were dominated by Streptococcus. Legionella was never the dominating genus and was always accompanied by other respiratory pathogens. Interestingly, sputum samples that were Legionella positive were inhabited by aquatic bacteria that have been observed in an association with amoeba, indicating that amoeba might have transferred Legionella from the drinking water together with its microbiome. This is the first study that demonstrates the sputum major bacterial commensals and pathogens profiles with regard to Legionella presence.


Subject(s)
Legionella/isolation & purification , Legionellosis/microbiology , Microbiota , Pneumonia/microbiology , Sputum/microbiology , Aged , Female , Humans , Legionella/genetics , Legionella pneumophila/genetics , Legionella pneumophila/isolation & purification , Legionellosis/complications , Male , Middle Aged , Pneumonia/complications
12.
Diagn Microbiol Infect Dis ; 87(2): 168-171, 2017 Feb.
Article in English | MEDLINE | ID: mdl-27889252

ABSTRACT

The aim of this study was to determine whether the route of extended-spectrum ß-lactamase (ESBL) transmission to hospitalized newborns was from their mothers during delivery. Neonatal intensive care unit (NICU) hospitalized newborns were sampled for ESBL presence by stool cultures on the first and fourth days of life. Mothers of ESBL-positive newborns were sampled for possible correlation detection. Bacteria isolates were molecularly identified and susceptibility tests for antibiotic agents were performed. Of the 225 newborns, 14 (6.2%) were ESBL positive, 10 (4.4%) were Escherichia coli positive, and 4 (1.7%) were Klebsiella pneumoniae positive. Among the 14 mothers of positive newborns, 13 (92.8%) were found ESBL positive and one mother of a newborn with E. coli carriage was found ESBL negative. Genes encoding for ESBL resistance were identified. Antibiotic sensitivity and resistance were tested. This study demonstrated that ESBL bacteria carrier neonates hospitalized in NICU may be a result of transmission from mother to baby during delivery.


Subject(s)
Carrier State/transmission , Enterobacteriaceae Infections/transmission , Escherichia coli/enzymology , Infectious Disease Transmission, Vertical , Klebsiella pneumoniae/enzymology , Peripartum Period , beta-Lactamases/metabolism , Adolescent , Adult , Carrier State/microbiology , Enterobacteriaceae Infections/microbiology , Escherichia coli/isolation & purification , Feces/microbiology , Female , Genotype , Humans , Infant, Newborn , Klebsiella pneumoniae/isolation & purification , Male , Microbial Sensitivity Tests , Molecular Typing , Phenotype , Young Adult
13.
BMC Infect Dis ; 16: 309, 2016 06 22.
Article in English | MEDLINE | ID: mdl-27334992

ABSTRACT

BACKGROUND: Clostridium difficile is the most common infectious etiology of nosocomial diarrhea. Fecal calprotectin (fc) is a sensitive marker of intestinal inflammation, found to be associated with enteric bacterial infections and inflammatory bowel disease. METHODS: We evaluated fc levels using a Chemiluminescent immunoassay method, in hospitalized patients with C. difficile infection (CDI) diagnosed by molecular stool examination and assessed correlation with virulent ribotype 027 strain infection, antibiotic susceptibility by gradient Etest strip performed on C. difficile colonies and clinical and laboratory measures of disease severity. Statistical analysis was performed for correlation of fc levels with clinical and laboratory parameters, disease severity and patient outcomes. RESULTS: Overall 29 patients with CDI were admitted at the Poria medical center in northern Israel, during June 2014-May 2015. Resistance to metronidazole was found in 3 (10.3 %) isolates and to vancomycin in 5 (17.2 %) isolates. Regarding patient outcomes, within 30 days of CDI diagnosis, recurrence of disease occurred in 10 (34.5 %) patients and 2 patients (6.9 %) died. Seven (24.1 %) isolates were C. difficile ribotype 027. Mean fc level was 331.4 µg/g (21-932). Higher fc levels were found in patients with C. difficile ribotype 027 (p < 0.0005). Fc levels were also correlated with elevated peripheral blood white cell count (p = 0.0007). A trend for higher fc levels was found in patients with a higher clostridium severity score index (p = 0.0633). No correlation was found between fecal calprotectin levels and age, sex, functional status, community versus hospital acquired CDI, antibiotic susceptibility, fever, and creatinine levels. CONCLUSIONS: Our study highlights the fact that fc has a potential role as a biomarker of disease severity and binary toxin producing ribotype associated disease.


Subject(s)
Clostridioides difficile/genetics , Clostridium Infections/microbiology , Feces/chemistry , Leukocyte L1 Antigen Complex/analysis , Adolescent , Adult , Aged , Aged, 80 and over , Biomarkers/analysis , Clostridioides difficile/isolation & purification , Clostridioides difficile/physiology , Clostridium Infections/metabolism , Clostridium Infections/physiopathology , Cross Infection , Drug Resistance, Bacterial , Female , Hospitalization , Hospitals , Humans , Immunoassay , Inflammation , Israel , Leukocytosis/blood , Male , Metronidazole , Middle Aged , Recurrence , Ribotyping , Severity of Illness Index , Vancomycin , Young Adult
15.
J Clin Lab Anal ; 30(5): 563-6, 2016 Sep.
Article in English | MEDLINE | ID: mdl-26666427

ABSTRACT

BACKGROUND: Identification of carbapenem-resistant Enterobacteriaceae (CRE) is complex and a major laboratory challenge; clinical cultures may diagnose only some of the CRE carriers among patients, thus it is crucial to perform asymptomatic carriage screening. MATERIALS AND METHODS:  We compare the efficacy of a rectal sample culture prior to enrichment with BHI (Brain Heart Infusion) Broth and following 18-24 h. All rectal samples were applied on CHROMagar KPC selective growth media and then seeded on MacConkey agar selective growth media with an applied disk of Imipenem antibiotic on top of the media, then inserted into enrichment BHI Broth. After 18-24 h incubation with enrichment media, all samples were applied again on this media. RESULTS: From the 2,245 rectal samples, CRE colonies were found in 96 (4.3%). Following enrichment with BHI Broth, CRE colonies were found in 111 (4.9%) CHROMagar KPC plates and 106 (4.7%) MacConkey agar. CONCLUSION: We were able to demonstrate that the number of CRE-positive results increased due to use of additional enrichment with BHI Broth. Therefore, we recommend applying this method of addition of liquid enrichment media as part of a culture protocol routine for CRE screening.


Subject(s)
Anti-Bacterial Agents/adverse effects , Carbapenems/adverse effects , Culture Media/pharmacology , Enterobacteriaceae Infections/drug therapy , Enterobacteriaceae/drug effects , Enterobacteriaceae/pathogenicity , Humans , Microbial Sensitivity Tests , Sensitivity and Specificity
17.
Am J Trop Med Hyg ; 93(2): 316-8, 2015 Aug.
Article in English | MEDLINE | ID: mdl-25962772

ABSTRACT

Acanthamoeba keratitis (AK), a potentially blinding infection of the cornea, is caused by a free-living protozoan. Culture and microscopic examination of corneal scraping tissue material is the conventional method for identifying Acanthamoeba. In this article, we compared several methods for AK diagnosis of 32 patients: microscopic examination using fluorescent dye, specific culture on growth media-non-nutrient agar (NNA), culture on liquid growth media-peptone yeast glucose (PYG), and TYI-S-33. AK was found in 14 patients. Thirteen of the specimens were found AK positive by fluorescence microscopic examination, 11 specimens were found AK positive on PYG growth media, and 9 specimens were found AK positive on TYI-S-33 growth media. Only five specimens were found AK positive on NNA growth media. Therefore, we recommend using fluorescence microscopy technique and culture method, especially PYG liquid media.


Subject(s)
Acanthamoeba Keratitis/diagnosis , Acanthamoeba/isolation & purification , Culture Media/chemistry , Culture Techniques/methods , Microscopy, Fluorescence/methods , Acanthamoeba/growth & development , Adult , Cornea/parasitology , Female , Humans , Male , Specimen Handling
18.
World J Gastroenterol ; 21(18): 5663-7, 2015 May 14.
Article in English | MEDLINE | ID: mdl-25987793

ABSTRACT

AIM: To prospectively examine the association between presence of Streptococcus bovis (S. bovis) in colonic suction fluid and the endoscopic findings on colonoscopy. METHODS: From May 2012 to March 2013, 203 consecutive patients who underwent colonoscopy for any reason were enrolled in the study. Exclusion criteria included: antibiotic use in the previous month, age younger than 18 years, and inadequate preparation for colonoscopy. The colonoscopy was performed for the total length of the colon or to the occluding tumor. The endoscopic findings were registered. Samples were obtained proximal to the colonoscopic part of the suction tube from each patient and sent to the clinical microbiology laboratory for isolation and identification of S. bovis. Samples were incubated in enrichment media with addition of antibiotic disks for inhibition of growth of Gram-negative rods. The samples were seeded on differential growth media; suspected positive colonies were isolated and identified with Gram staining, catalase, and pyrrolidonyl arylamidase tests, and further identified using a VITEK2 system. Statistical analyses were performed using the Student's t and χ(2) tests. RESULTS: Of the 203 patients recruited, 49 (24%) patients were found to be S. bovis carriers; of them, the endoscopic findings included: 17 (34.7%) cases with malignant tumors, 11 (22.4%) with large polyps, 5 (10.2%) with medium-sized polyps, 6 (12.2%) with small polyps, 4 (8.1%) with colitis, and 6 (12.2%) normal colonoscopies. Of 154 patients found negative for S. bovis, the endoscopic findings included: none with malignant tumors, 9 (5.8%) cases with large polyps, 11 (7.1%) with medium-sized polyps, 26 (16.9%) with small polyps, 7 (4.5%) with colitis, and 101 (65.6%) normal colonoscopies. S. bovis (Gram-positive coccus) is considered part of the normal intestinal flora. There is an association between S. bovis bacteremia and colonic neoplasia. It is not well understood whether the bacterium has a pathogenetic role in the development of neoplasia or constitutes an epiphenomenon of colorectal neoplasms. There was a clear relationship between positivity for S. bovis in colonic suction fluid and findings of malignant tumors and large polyps in the colon. CONCLUSION: There is an association between S. bovis bacteremia and malignant colonic lesions; this should prompt for development of a reliable screening method for advanced colonic lesions.


Subject(s)
Colon/microbiology , Colonic Neoplasms/microbiology , Colonic Polyps/microbiology , Streptococcal Infections/microbiology , Streptococcus bovis/isolation & purification , Adult , Aged , Aged, 80 and over , Chi-Square Distribution , Colon/pathology , Colonic Neoplasms/pathology , Colonic Polyps/pathology , Colonoscopy , Disk Diffusion Antimicrobial Tests , Female , Humans , Male , Middle Aged , Predictive Value of Tests , Prospective Studies , Streptococcal Infections/diagnosis , Tumor Burden , Young Adult
19.
Am J Infect Control ; 43(7): e19-22, 2015 Jul 01.
Article in English | MEDLINE | ID: mdl-25920705

ABSTRACT

BACKGROUND: Catheter-associated candiduria is a common clinical finding in hospitalized patients, especially in the intensive care unit. The objective of this study was to obtain demographic and clinical data regarding the prevalence of Candida spp in catheterized in-patients and the medical interventions provided to these patients in a northern Israeli hospital between 2011 and 2013. METHODS: Isolation and identification of microorganisms were performed on 1,408 urine culture samples 48 hours after catheter insertion. Antifungal Etest susceptibility tests were carried out on every Candida-positive urine sample. Demographic and clinical data were gathered to determine risk factors and medical interventions. RESULTS: Candiduria was detected in 146 catheterized in-patients out of the 1,408 patients included in this study. C albicans was detected in most cases (69.1%). Fever was observed in 52 (35.61%) patients, and leukocyturia was observed in 48 cases (32.87%). Diabetes mellitus was associated with C albicans candiduria. There were 93 patients (63.69%) who did not receive any medical intervention for their candiduria. CONCLUSION: Candida is the second leading pathogen causing catheter-associated urinary tract infection or asymptomatic colonization, whereas previous studies showed Candida as the third leading pathogen. Clinical signs and symptoms, such as fever and laboratory tests, cannot distinguish between asymptomatic colonization and infection. Because the management of catheter-associated candiduria is still controversial, additional studies should be carried out.


Subject(s)
Antifungal Agents/therapeutic use , Candida/drug effects , Candidiasis/drug therapy , Catheter-Related Infections/drug therapy , Urinary Tract Infections/drug therapy , Antifungal Agents/pharmacology , Candida/isolation & purification , Candidiasis/epidemiology , Candidiasis/microbiology , Catheter-Related Infections/epidemiology , Catheter-Related Infections/microbiology , Disk Diffusion Antimicrobial Tests , Female , Hospitals , Humans , Israel/epidemiology , Male , Middle Aged , Prevalence , Risk Factors , Urinary Tract Infections/epidemiology , Urinary Tract Infections/microbiology
20.
J Med Microbiol ; 64(6): 642-645, 2015 Jun.
Article in English | MEDLINE | ID: mdl-25878170

ABSTRACT

Helicobacter pylori infection represents a key factor in the aetiology of various gastrointestinal diseases. H. pylori infection diagnosis is generally achieved using both invasive (e.g. biopsy of the gastric epithelium) and non-invasive methods. Therefore, cultivation on a growth medium becomes complex. Trypsin is a proteinase enzyme that plays a role in an early stage of tissue digestion. In this study, we used trypsin in order to improve the diagnostic sensitivity of the H. pylori cultivation technique. We used 46 duplicate antrum biopsy specimens, divided into trypsin-treated and non-treated groups. The tissues were seeded on a selective H. pylori growth agar medium. We demonstrated that the classic H. pylori culture technique misses the growth of a large number of H. pylori colonies. Significantly more colonies were found in the trypsin-treated specimens group.


Subject(s)
Bacteriological Techniques/methods , Helicobacter pylori/isolation & purification , Specimen Handling/methods , Trypsin/metabolism , Adolescent , Adult , Biopsy , Gastric Mucosa/microbiology , Helicobacter Infections/diagnosis , Humans , Sensitivity and Specificity
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