Antimicrobial-resistant Klebsiella species isolated from free-range chicken samples in an informal settlement

Introduction

Sub-therapeutic doses of antimicrobial agents are administered routinely to poultry to aid growth and to prevent disease, with prolonged exposure often resulting in bacterial resistance. Crossover of antibiotic resistant bacteria from poultry to humans poses a risk to human health.

Material and methods

In this study, 17 chicken samples collected from a vendor operating in an informal settlement in the Cape Town Metropolitan area, South Africa were screened for antimicrobial-resistant Gram-negative bacilli using the Kirby Bauer disk diffusion assay.

Results

In total, six antibiotics were screened: ampicillin, ciprofloxacin, gentamicin, nalidixic acid, tetracycline and trimethoprim. Surprisingly, Klebsiella ozaenae was identified in 96 and K. rhinoscleromatis in 6 (n=102) of the samples tested. Interestingly, ∼40% of the isolated Klebsiella spp. showed multiple resistance to at least three of the six antibiotics tested.

Conclusions

Klebsiella ozaenae and K. rhinoscleromatis cause clinical chronic rhinitis and are almost exclusively associated with people living in areas of poor hygiene.     

Procalcitonin levels in salmonella infection

Aim:

Procalcitonin (PCT) as a diagnostic marker for bacteremia and sepsis has been extensively studied. We aimed to study PCT levels in Salmonella infections whether they would serve as marker for early diagnosis in endemic areas to start empiric treatment while awaiting blood culture report.

Materials and Methods:

BACTEC blood culture was used to isolate Salmonella in suspected enteric fever patients. Serum PCT levels were estimated before starting treatment.

Results:

In 60 proven enteric fever patients, median value of serum PCT levels was 0.22 ng/ml, values ranging between 0.05 and 4 ng/ml. 95% of patients had near normal or mild increase (<0.5 ng/ml), only 5% of patients showed elevated levels. Notably, high PCT levels were found only in severe sepsis.

Conclusion:

PCT levels in Salmonella infections are near normal or minimally increased which differentiates it from other systemic Gram-negative infections. PCT cannot be used as a specific diagnostic marker of typhoid.     

Carbapenem-resistant Acinetobacter ventilator-associated pneumonia: Clinical characteristics and outcome

Objective:

To study the clinical characteristics and 28-days mortality in patients with ventilator-associated pneumonia (VAP) due to carbapenem-resistant Acinetobacter (CRA).

Design:

Retrospective, observational, cohort study.

Setting:

Intensive care unit (ICU) of a university hospital.

Materials and Methods:

Microbiologically confirmed VAP due to CRA infection.

Intervention:

None.

Results:

Out of 87 patients with VAP due to CRA, 60 (69%) were male; whose median age was 51 years; 73 (84%) patients were medical; 26 (30%) had history of hospitalization in last 3 months; median acute physiology and chronic health evaluation (APACHE) II was 15 and median SOFA 9 at admission; primary reason for ICU admission was respiratory failure (34%); 46 (53%) patients had more than 2 organ failure at ICU admission; median length of ICU stay was 19 days; 66 (76%) patients need vasoactive agents during ICU stay, whereas 55 (63%) patients had renal failure; median duration of mechanical ventilation was 17 days; 22 (25%) patients had acute respiratory distress syndrome (ARDS) during ICU stay; 72 (83%) patients had exposure to carbapenem before inclusion in the study; 33 (38%) patients had same organism at other sites. In the follow-up, 47 (54%) patient survived at 28 days after having VAP; whereas only 40 (46%) patients were discharged from the hospital.

Conclusions:

CRA-VAP has high crude mortality. Advanced age; severity of illness and presence of pneumonia at ICU admission; and presence of shock, ARDS and renal failure have impact on outcome in these patients.     

Carbapenem resistant Gram-negative bacteremia in an Indian intensive care unit: A review of the clinical profile and treatment outcome of 50 patients

Background:

Growing antimicrobial resistance and limited therapeutic options to treat carbapenem-resistant bacteremia prompted us to evaluate the clinical outcomes associated with healthcare-associated bacteremia.

Methods:

This was a retrospective observational study of carbapenem-resistant Gram-negative bacteremia performed at a tertiary care facility in Chennai, India between May 2011 and May 2012.

Results:

In our study, patients had mean 11.76 days of intensive care unit (ICU) care and mean time to onset of bacteremia was 6.4 days after admission. The commonest organism was Klebsiella pneumoniae (44%). Patients with combination treatment had lower mortality (44.8%) compared with colistin monotherapy (66.6%); (P = 0.35).

Conclusion:

Carbapenem resistant bacteremia is a late onset infection in patients with antibiotic exposure in the ICU and carries a 30 days mortality of 60%; K. pneumoniae is the most common organism at our center. Two drug combinations appear to carry a lower mortality compared with monotherapy.     

Extract from Aronia melanocarpa fruits potentiates the inhibition of platelet aggregation in the presence of endothelial cells

Introduction

Some polyphenolic compounds extracted from Aronia melanocarpa fruits (AM) have been reported to be cardioprotective agents. In this study we evaluated the ability of AM extract to increase the efficacy of human umbilical vein endothelial cells (HUVECs) to inhibit platelet functions in vitro.

Material and methods

This study encompasses two models of monitoring platelet reactivity: optical aggregation and platelet degranulation (monitored as the surface CD62P expression) in PRP upon the stimulation with ADP.

Results

We observed that only at low concentrations (5 µg/ml) did AM extract significantly improve antiplatelet action of HUVECs towards ADP-activated platelets in the aggregation test.

Conclusions

It is concluded that the potentiating effect of AM extract on the endothelial cell-mediated inhibition of platelet aggregation clearly depends on the used concentrations of Aronia-derived active compounds. Therefore, despite these encouraging preliminary outcomes on the beneficial effects of AM extract polyphenols, more profound dose-effect studies should certainly be considered before the implementation of Aronia-originating compounds in antiplatelet therapy and the prevention of cardiovascular diseases.     

<Emphasis Type="Italic">Celastrus orbiculatus</Emphasis> extracts induce apoptosis in mTOR-overexpressed human hepatocellular carcinoma HepG2 cells

Background

Celastrus orbiculatus (Celastraceae) are used as traditional Chinese medicine to treat inflammation and cancer. This study aims to evaluate the effect of Celastrus orbiculatus extract (COE) on the apoptosis in human hepatic carcinoma HepG2 cells with mTOR overexpression.

Methods

The stable expression of mTOR in HepG2 cells (HepG2/mTOR⁺) were established by lipofectin transfection of GV238-mTOR recombinant plasmids and further antibiotic selection. Human hepatic carcinoma HepG2/mTOR⁺ cells were treated with different concentrations (20, 40, 80, 160, and 320?μg/mL) of COE for 24?h. The cell proliferation upon COE treatment was detected by MTT. Apoptosis was measured by Flow Cytometry. The activity of mTOR signaling pathway was detected by Western Blotting.

Results

COE significantly inhibited the proliferation of HepG2/mTOR⁺ cells. The expression levels of Bax and Caspase-3 protein were increased in the HepG2/mTOR⁺ cells in a dose-dependent manner. The proteins expression of Bcl2, Bcl-2?L12, mTOR, phospho-mTOR, 4EBP1, phospho-4EBP1, P70S6k, and phospho-P70S6k in HepG2/mTOR⁺ cells were reduced in dose-dependent manners. Furthermore, COE and mTOR inhibitor rapamycin (RAPA) synergistically induced apoptosis in HepG2/mTOR⁺ cells by regulating apoptosis-related proteins and inhibiting mTOR signaling pathways.

Conclusion

COE could inhibit the proliferation of HepG2/mTOR⁺ cells, and induce the cell apoptosis. The mechanisms may be related to the regulation of the expression of Bcl-2, Bcl-2?L12, and mTOR signaling pathways. These data suggest that COE may be a potential treatment for human hepatocellular carcinoma.

     

Nosocomial candiduria in chronic liver disease patients at a hepatobilliary center

Background:

Nosocomial urinary tract infections (UTIs) are common in catheterized patients. Fungal UTI has become an important nosocomial problem over the past decade. The microbiology of candiduria is rapidly evolving and new trends are being reported.

Aims:

To study the microbiological trends and antifungal resistance profile of Candida in urine of catheterized chronic liver disease (CLD) patients at a super specialty hepatobiliary tertiary-care center.

Materials and Methods:

urine samples were collected by sterile technique, processed by semi-quantitative method as per the standard protocols. Direct microscopic examination of urine sample was also done to look for the presence of pus cells, red blood cells, casts, crystals or any bacterial or fungal element.

Result:

A total of 337 yeast isolates were obtained from catheterized patients, non-albicans Candida spp. emerged as the predominant pathogen and was responsible for 67.06% of nosocomial fungal UTI. Candida tropicalis accounted for 34.71% of the cases, whereas Candida albicans grew in 32.93%, Candida glabrata 16.32%, rare Candida spp. Nearly 11.5% (Candida hemolunii to be confirmed by molecular methods). Antifungal sensitivity varied non-albicans species except C. tropicalis, Candida parapsilosis were more often resistant to antifungal drugs.

Conclusion:

Nosocomial Candida UTIs in CLD patients is common, due to the cumulative pressure of contributing factors such as urinary instrumentation and prolonged use of broad-spectrum antibiotics. Non-albicans Candida were found to outnumber C. albicans in catherized CLD patients. Risk of strain persistence is also higher with non-albicans Candida. Thus, species identification and susceptibility testing is a must for appropriate management of such patients.     

A novel ropivacaine-loaded in situ forming implant prolongs the effect of local analgesia in rats

Introduction

Prolonged postoperative analgesia cannot be achieved by a single injection of local anesthetic solution. The objective of this study was to optimize the formulation of a ropivacaine hydrochloride (Ropi-HCl) loaded in situ forming implant (ISI) by addition of different co-solvents, and evaluate the in vitro release of Ropi-HCl, and the analgesic effect and toxicity of the optimized formulation in rats.

Material and methods

Triacetin (TA), benzyl benzoate (BB) and polyethylene glycol 400 (PEG 400) were used as additives and added to the solvent of N-methyl-2-pyrrolidone (NMP). Drug release to the surface and inner structural properties of the formed implant were evaluated by scanning electron microscopy (SEM). The analgesic effect was determined by injection near the rat sciatic nerve.

Results

The solvent system added with TA or BB significantly decreased the burst release, whereas PEG 400 increased the Ropi-HCl burst release from the formulation. Over 70% of the incorporated Ropi-HCl was released from all formulations in 14 days in the in vitro assay. The SEM showed that the surface of NMP-BB formulation was less porous and more homogeneous, compared with the other formulations. Compared with Ropi-HCl injection, the optimized formulation (NMP-BB) significantly prolonged the analgesic effect in 48 h (p < 0.05), with a mild degree of motor block from 3 h to 12 h. Histological evaluation of the injection site revealed only mild inflammatory infiltration without obvious pathological nerve alterations.

Conclusions

The biodegradable Ropi-HCl-loaded ISI system with NMP-BB may prove to be an attractive and safe alternative for the delivery of parenteral local anesthetics to prolong pain relief.     

Prevalence and antimicrobial susceptibility pattern of methicillin-resistant Staphylococcus aureus in Assam

Aims:

Methicillin-resistant Staphylococcus aureus (MRSA) has become a serious problem in intensive care units, because of development of multiresistance, and also intrinsic resistance to β-lactam antibiotics. The present study was carried out to investigate the prevalence of MRSA and their rate of resistance to different antistaphylococcal antibiotics.

Materials and Methods:

Between January 2007 and February 2008, the clinical specimens submitted at the microbiology laboratory were processed and all S. aureus isolates were included in this study. All isolates were identified morphologically and biochemically by standard laboratory procedures and antibiotic susceptibility pattern was determined by modified Kirby Bauer disc diffusion method.

Results:

Methicillin resistance was observed in 34.78% of isolates, of which 37.5% were found to be resistant to all commonly used antibiotics. In MRSA isolates, 50% had constitutive resistance, 9.38% had inducible MLS_B resistance and 18.75% had MS phenotype.

Conclusions:

There is a progressive increase in MRSA prevalence in the country but the present rate is still low in comparison to values found in some other institutes. The rate of inducible MLS_B resistance was also lower in comparison with findings from other parts of the country.     

Clinical efficacy and safety of colistin treatment in patients with pulmonary infection caused by Pseudomonas aeruginosa or Acinetobacter baumannii: a meta-analysis

Introduction

The aim of this study was to evaluate the efficacy and safety of colistin treatment in patients with pulmonary infection caused by Pseudomonas aeruginosa or Acinetobacter baumannii.

Material and methods

The relevant studies were identified through a search of public databases including PubMed, MEDLINE and EMBASE up to December 2012. A meta-analysis was conducted to compare the clinical response, mortality and renal damage of colistin (colistin group) versus other effective antibiotics (control group). The odds ratio (OR) was chosen as the effect size.

Results

A total of 9 studies were eventually identified. The result of the meta-analysis showed that the pooled OR of clinical response was 1.24 (95% CI = 0.68–2.27, p > 0.05) for patients in the colistin group versus the control group, indicating no significant difference in efficacy between colistin and control groups. Similar results were obtained by the further subgroup meta-analyses by sample size, research year, ethnicity and study method. Treatment with colistin versus other agents did not affect hospital mortality (OR = 1.05, 95% CI = 0.58–1.89, p > 0.05) or renal damage (OR = 1.25, 95% CI = 0.78–2.00, p > 0.05). The combined estimate of our analysis was strong across multiple sensitivity analyses and without significant publication bias.

Conclusions

Our results suggest that colistin may be as efficacious and safe as standard antibiotics for the treatment of pulmonary infection.     

Examination of the FLT3 and NPM1 mutational status in patients with acute myeloid leukemia from southeastern Poland

Introduction

Acute myeloid leukemia (AML) is a genetically heterogeneous disease at both the cytogenetic and molecular levels. In AML cells many chromosomal aberrations are observed, some of them being characteristic of a particular subtype of patients, and others being less significant. Besides chromosomal abnormalities, the leukemic cells can have a variety of mutations involving individual genes. The aim of this work was to investigate the frequencies of molecular alterations with the focus on FLT3-ITD and NPM1 mutations in AML patients of different age groups living in a southeastern region of Poland.

Material and methods

The study group comprised 50 consecutive AML patients. We analyzed bone marrow samples by conventional cytogenetics. Cytogenetic evaluation in selected cases was complemented by the FISH technique. The internal tandem mutation in the FLT3 gene was identified using polymerase chain reaction (PCR), and the NPM1 mutation was assessed by direct nucleotide sequencing.

Results

The studies using classical cytogenetics showed chromosomal aberrations in 32 (64%) patients. In 18 cases no changes in the karyotype were found by conventional karyotyping. FLT3-ITD mutation was detected in 4 (8%) patients and mutation of NPM1 in 3 patients with AML (6%).

Conclusions

The incidence of both mutations in our study group was lower than described elsewhere. We have confirmed that FLT3-ITD occurred more commonly in older patients and it was associated with shorter overall survival. By contrast, mutation of exon 12 of the NPM1 gene seems to be a good prognostic factor in AML patients with normal karyotype.     

Comparison of the GenoType? MTBC Molecular Genetic Assay with culture methods in the diagnosis of tuberculosis

Introduction

Clinical samples from 433 patients pre-diagnosed with tuberculosis in Konya, Turkey, were investigated prospectively to compare the GenoType^® MTBC test (GenoType^® MTBC) with conventional “gold standard” culture methods.

Material and methods

Lowenstein Jensen (LJ) and Mycobacteria Growth Indicator Tube (MGIT)-960 culture methods and GenoType^® MTBC were performed together.

Results

Mycobacterium tuberculosis (M. tuberculosis) detection rates were 16.2% by culture methods, 15.4% by GenoType^® MTBC, and 6% by acid-fast bacilli microscopy. The LJ or MGIT-960 with GenoType^® MTBC detected M. tuberculosis in 12 samples each that were negative according to the other culture method alone. Among 70 M. tuberculosis-positive samples, detection rates were 37% (26/70) by microscopy and 82.8% (58/70) by LJ and MGIT-960, but 95.7% (67/70) by GenoType^® MTBC.

Conclusions

GenoType^® MTBC may be used as a beneficial adjunct test to culture methods for the detection of M. tuberculosis.     

Bioactivity-Guided Isolation of Anticancer Agents from Bauhinia Kockiana Korth

Background

Flowers of Bauhinia kockiana were investigated for their anticancer properties.

Methods

Gallic acid (1), and methyl gallate (2), were isolated via bioassay-directed isolation, and they exhibited anticancer properties towards several cancer cell lines, examined using MTT cell viability assay. Pyrogallol (3) was examined against the same cancer cell lines to deduce the bioactive functional group of the phenolic compounds.

Results

The results showed that the phenolic compounds could exhibit moderate to weak cytotoxicity towards certain cell lines (GI₅₀ 30 – 86 µM), but were inactive towards DU145 prostate cancer cell (GI₅₀ > 100 µM).

Conclusion

It was observed that pyrogallol moiety was one of the essential functional structures of the phenolic compounds in exhibiting anticancer activity. Also, the carboxyl group of compound 1 was also important in anticancer activity. Examination of the PC-3 cells treated with compound 1 using fluorescence microscopy showed that PC-3 cells were killed by apoptosis.     

The anti-tumor effect of a polypeptide extracted from Tegillarca granosa Linnaeus on renal metastatic tumor OS-RC-2 cells

Introduction

Renal cell carcinoma is a type of malignant tumor often diagnosed in the urinary system. The aim of this study is to evaluate the anti-tumor effects and mechanisms of a polypeptide, Haishengsu (TG-1), with different concentrations (125, 250, 500 mg/kg) on renal metastatic tumor OS-RC-2 cells.

Material and methods

We first established the renal metastatic tumor model. After being administered with TG-1, the weight of tumors was measured and the microstructural changes of renal carcinoma OS-RC-2 cells were compared using transmission electron microscopy before and after the therapy. The Ki67 expression in renal carcinoma OS-RC-2 cells was analyzed by RT-PCR and downstream signal molecule caspase-3 was measured by Western blot assay.

Results

After treatment with different doses of TG-1, the tumor weights in the positive control group and experimental groups were smaller than those in the untreated control group, suggesting that TG-1 could effectively inhibit tumor growth in mice. The transmission electron microscopy and flow cytometry results indicated that TG-1 induces tumor cell apoptosis (p < 0.05). The tumor cells exhibited polymorphism changes and chromatin edge clustering. TG-1 also inhibited Ki67 expression and promoted caspase-3 expression in the tumor significantly (p < 0.05).

Conclusions

TG-1 inhibits the growth of the tumor and induces apoptosis of the tumor cells by inducing caspase-3 expression. The results provide a basis for clinical application of TG-1 in the future.     

The Antibacterial Activity of Traditionally Used Salvadora Persica L. (Miswak) and Commiphora Gileadensis (Palsam) in Saudi Arabia

Background

Nowadays there is a need to find naturally occurring substances from plants with antimicrobial activity as an alternative to available used antibiotics.

Materials and Methods

Salvadora persica (miswak) and Commiphora gileadensis were collected, dried and extracted with either methanol or warm water and the obtained extracts were assessed for their antibacterial activity against 5 different genera of bacteria using agar well diffusion method. The tested bacteria included some human pathogens.

Results

The obtained extracts exhibited considerable inhibitory effects against all the tested bacteria with various degrees of growth inhibition. It was shown that methanol extract was more effective compared to water extracts. The minimum inhibitory concentrations (MIC) of the methanol extracts ranged from 50–100 °g/ml. No toxicity was found using Artimia salina as test organism and no antitumor activity against Ehrlich ascites carcinoma.

Conclusion

S. persica and C. gileadensis showed moderate to high inhibitory activity on pathogenic bacteria with no toxicity and can be used traditionally as alternative medicine     

Antimicrobial susceptibility of Helicobacter pylori isolates from Lower Silesia,Poland

Introduction

In recent years the failure of standard therapy for Helicobacter pylori infections has been observed, which results primarily from the increasing resistance of H. pylori strains to antibiotics. The aim of the study was to estimate the prevalence of antimicrobial resistance of H. pylori strains isolated from adult symptomatic patients with primary infection in the Lower Silesia Region in Poland.

Material and methods

One hundred and seventy-eight adults aged 19–89 years with dyspeptic symptoms suggesting gastroduodenal pathology were enrolled in the study. The study was performed in the years 2008–2011. Fifty H. pylori strains were isolated from gastric biopsy samples of examined patients. Antimicrobial susceptibility to 6 drugs (amoxicillin (AM), clarithromycin (CH), metronidazole (MZ), tetracycline (TC), levofloxacin (LEV), and rifabutin (RB)) was tested by the gradient-diffusion method (E-test method).

Results

The incidence of H. pylori infection among examined patients was 35%. From 50 isolated H. pylori strains, 24% showed resistance to CH, 42% to MZ and 8% to LEV alone. Multidrug resistance was detected in 26% of strains, whereas 20% of isolates were resistant to MZ and CH. Examined strains were fully susceptible to AM, TC and RB.

Conclusions

Resistance to clarithromycin strains isolated from adults of the Lower Silesia Region in Poland is high and is almost always associated with resistance to metronidazole (CH + MZ). It is necessary to continuously monitor H. pylori resistance to drugs used in therapy, especially to clarithromycin. Verification of the existing recommendations of eradication therapy is also needed.     

Effects of Ixeris Chinensis (Thunb.) Nakai Boiling Water Extract on Hepatitis B Viral Activity and Hepatocellular Carcinoma

Background

Hepatitis B virus (HBV) infection and hepatocellular carcinoma are major diseases that affect the Taiwanese population. Therefore, the development of an alternative herbal medicine that can effectively treat these diseases is a research target. In this study, we tested Ixeris Chinensis (Thunb.) Nakai boiling water extract (ICTN BWE) in vitro and analysed its effects on the HBV and liver cancer.

Materials and Methods

We used a human liver cancer cell line (Hep3B, a cell line that continuously secretes HBV particles into a medium) as an experimental model for the screening of various ICTN BWE concentrations and their effects on the HBV in vitro.

Results

Our results showed that 75 µg/mL ICTN BWE downregulated the relative expression of the hepatitis B virus surface antigens (HBsAg) to 77.1%. Using the human liver cancer cell lines HuH-7 and HepG2, and 3-(4,5-dimethylthiazol-zyl)-2,5-diphenyl tetrazolium bromide (MTT) and tumour clonogenic assays, we then showed that ICTN BWE inhibits hepatocellular carcinoma growth.

Conclusion

Fluorescent microscopy of DAPI(4′,6-Diamidino-2-phenylindole)-stained nuclei and DNA fragmentation assays confirmed the inhibitory effects of ICTN BWE on liver tumour cell growth through induction of apoptosis.     

Three-dimensional analysis of cervical spine segmental motion in rotation

Introduction

The movements of the cervical spine during head rotation are too complicated to measure using conventional radiography or computed tomography (CT) techniques. In this study, we measure three-dimensional segmental motion of cervical spine rotation in vivo using a non-invasive measurement technique.

Material and methods

Sixteen healthy volunteers underwent three-dimensional CT of the cervical spine during head rotation. Occiput (Oc) – T1 reconstructions were created of volunteers in each of 3 positions: supine and maximum left and right rotations of the head with respect to the bosom. Segmental motions were calculated using Euler angles and volume merge methods in three major planes.

Results

Mean maximum axial rotation of the cervical spine to one side was 1.6° to 38.5° at each level. Coupled lateral bending opposite to lateral bending was observed in the upper cervical levels, while in the subaxial cervical levels, it was observed in the same direction as axial rotation. Coupled extension was observed in the cervical levels of C5-T1, while coupled flexion was observed in the cervical levels of Oc-C5.

Conclusions

The three-dimensional cervical segmental motions in rotation were accurately measured with the non-invasive measure. These findings will be helpful as the basis for understanding cervical spine movement in rotation and abnormal conditions. The presented data also provide baseline segmental motions for the design of prostheses for the cervical spine.