Nina Pastukh

Association of Streptococcus bovis presence in colonic content with advanced colonic lesion

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 Students 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.

The antimicrobial susceptibility of Helicobacter pylori in strains isolated from children in Israel

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.

Peripartum maternal transmission of extended-spectrum β-lactamase organism to newborn infants

The aim of this study was to determine whether the route of extended-spectrum β-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.

Antimicrobial susceptibility of Clostridium difficile isolates in Israel

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.

In Vitro Antileishmanial Activity of a Black Morel, Morchella importuna (Ascomycetes)

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.

Comparison of sputum microbiome of legionellosis-associated patients and other pneumonia patients: indications for polybacterial infections

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.

GenoType® HelicoDR test in comparison with histology and culture for Helicobacter pylori detection and identification of resistance mutations to clarithromycin and fluoroquinolones

There are several methods for Helicobacter pylori infection diagnosis.

Colony Forming Unites - Endothelial Progenitor Cells (CFU-EPCs) A surrogate marker for diabetic retinopathy and high cardiovascular mortality rate

Purpose: Diabetic retinopathy is a risk factor for increased cardiovascular death. Our purpose was to find a significant difference in levels of endothelial progenitor cells (EPCs) in the peripheral blood of patients at different stages of diabetic retinopathy. Design: A prospective study. Colony forming units of endothelial progenitor cells (CFU-EPCs) in peripheral blood were counted. 40 subjects were enrolled (10 healthy [41±8 y], 10 type 2 diabetes mellitus (T2DM) [64±12 y] without retinopathy, 10 T2DM patients [62±26 y] with non-proliferative retinopathy (NPDR), 10 T2DM patients [66±9 y] with proliferative retinopathy (PDR)). The study was approevd by the ethics committee of the hospital and every subject signed a soncent form before enrollment. Methods: Growing CFU-EPCs was by the Hills EPCs protocol. Blood was drawn early in the morning and was processed within 1 hour. Mononuclear cells were separated and cultured on fibronectin-coated plates with EndoCult medium (StemCell technologies, Vancouver BC Canada) for 5 days. CFU-EPCs were counted on day 5 (an average of 8 wells). Results: Healthy subjects had 36±8 CFU-EPCs, patients without retinopathy had 13±12 CFU-EPCs (p<0.01), patients with NPDR 22±26 CFU-EPCs (p=NS), and 2±2 CFU-EPCs in patients with PDR (p<0.01). A significant difference was found between patients with PDR and with NPDR (p<0.05). Conclusions: CFU-EPCs are inhibited in T2DM patients with DPR. Levels of CFU-EPCs may be used as a surrogate biologic marker for severity of diabetic retinopathy and for cumulative vascular risk.Purpose: Representing the algorithm of periorbital area rejuvenation Materials: As part of the research, 1216 eyes of 608 patients were included. According to patients’ complaints and evaluating their situation, treatments were carried out as medical and surgical. Treatment was not applied to 75 patients that their request and the cure were not compromise. Patients were followed approximately 11,3 months (1-32 months). The study is evaluated according to patients’ reason of request, their general characteristics and the treatments that are applied. Findings: Age average of 608 patients who are applied to my clinic with the intent of periorbital area rejuvenation and enhancing eyelid deformity between the dates of March 2014-January 2017 is 38. 81,25% patients were female and 18,75% was male. 538 patients (88,5%) were applied medical treatment. 260 patients (42,8%) got surgical treatment and to 31,3% patients, combined treatment was applied. The treatment that I applied and the complaints of 89% patients harmonized with each other. 11% of patients were not treated since their complaints and their treatment request were not corresponded to each other. Result: The most important phase is pre-treatment to become successful in the treatment. First thing to do is listening and understanding the patient carefully, because the problem we observe and patient’s complaint may not be the same. In this case, no matter how successful the treatment is, we can not satisfy the patient. Sometimes, patient has unrealistic expectations. When such a patient like this is encountered, it must be kept away from him or her and the treatment should not be applied. Volume 2 | Issue 2 | 1 of 4 Keyword: Periorbital Area, Rejuvenation, Surgical, Medical Aesthetics, Eyelid Blepharoplasty. Introduction Periorbital area is the most frequently-consulted and watched out area in bilateral relations so it has gained much more importance in recent years. It is possible to be looked healthier and younger with lots of practice applied around eye. With this reason, more and more patients apply to clinics to look better everyday. As an ophthalmologist, I would like to mention about the treatments that I provide to make my patients look better, younger and healthier. Most people want to look better with the improvement of technology, increasement of using social media and importance of visual quality. With this aim in their minds, patients consult health institutions. In my clinic, consultation request is generally for periorbital area. The mistreatments in the periorbital area which has an important function as seeing cause vision loss. This situation could arise from unrealistic expectations of the patient or insufficiency in the health experience of health center. Also, the misunderstanding between the patient and the doctor may cause this problem [1]. I think that ophthalmologists should actively participate in the treatments of periorbital area. After examining the patient in detail, the treatment should be explained loud and clear. It can be repeated until he or she understands the approach clearly. While the treatment is thought as surgical and medical, it can be also combined. It should be decided according to the request of patient and its satisfying outcome. In the patient who complains about upper droopy eyelid, contexts of the eyelid and whether they include additional pathology or not should be examined. Eyelid, eyebrow and eyelash ptosis should be evaluated. Whether excised area includes fat package or not without skin and orbicularis should be reviewed. In the patient with the request of low eyelid blepharoplasty, the trace of entropion and ectropion should be looked. With the snap back test, the laxity of low eyelid should be examined. In this part, the existence of scleral show helps us. Additionally, cheek looseness and laxity in the malar pad should be evaluated [2]. In the patient, medical treatment can be thought not surgical or in addition to that surgery, medical treatment can be planned. In this part, the treatments that help us are botox in eyebrow lid andL Eye” syndrome or amblyopia is a common phenomenon scattered throughout the human population. It is estimated that around 5% of the human population suffers from this syndrome along with diplopia, and refractive squint present in many or rather most of the cases. It is quite possible that other animal species that depend on stereoscopic vision for their survival are also prone to this condition in the same frequency as the human population. During prehistoric and ancient times, this condition did not interfere with human survival and this is the reason of its prevalence among the human population even in the present times. As it is present among 5% of the human population, it is no doubt a genetic condition. It has presented itself in individuals that have a genetic history of this condition in the maternal or paternal lineages. This condition is a major liability for the sufferer in the modern world. During ancient times, when man started as a hunter-gatherer and until the human world did not depend on the written word, this condition was a benign syndrome, possibly without any competitive disadvantage. That is the reason this condition has survived into the modern world. In the modern world, the major disadvantage is in field of academics. Children suffering from this condition are at a disadvantage due to their inability to study like other normal children. In the amblyopic patients, having diplopia and refractive squint near stereoscopic vision is disrupted during reading and other near vision activities. Two different images that are created in the visual cortices do not overlap. The glasses force the visual cortices to fuse the two different images into one which otherwise is impossible and makes amblyopia comorbid with refractive squint a difficult malady to correct with present treatment options. These glasses are based on the neurobiology of vision. How stereoscopic vision is processed by the brain and the understanding of underlying cause that produces this deficit in the brain of individuals afflicted with this disorder is also conceptualized in this treatment option. Why 3D optics is helpful in correcting this disorder in the brain is also hypothesized in this paper. These glasses should only be used for near vision activities like reading through a book but not for reading through a computer screen or watching television, as no diplopia is present when words or visuals are seen on an illuminated background, although amblyopic patients who suffer headache while using a computer shall benefit immensely from this procedure. This method will correct the visual deficit and convergence insufficiency with 100% positive results and shall improve the academic capabilities of children suffering from amblyopia and refractive squint who complain of diplopia and headache while reading. These glasses will also correct the binocular acuity of individuals suffering from amblyopia. This can be done by breaking the suppression of visual signals sent by the amblyopic eye to the brain. Prolonged use of these glasses will train the brain to make use of signals and images created in both the visual cortices into one. Hence, the brain will be forced to utilize visual signals from both the eyes due to its neurophysiology.Purpose: To evaluate the results of patients with complaint of under eye hollowness and dark circles and application of under eye hyaluronic acid filler, comparison of the cannula and injection techniques. Method: Under eye hyaluronic acid filler was applied to the patients who consult to the clinic with complaint of hollows and dark circles under eye between March 2014 and December 2016. Application method was evaluated according to the success of results, satisfaction and complications. Findings: 356 eyes of 178 patients were included to the research. There were 148 (83%) female and 30 (17%) male patients. Average age was 33 (age range 19 60). It was defined that the most frequently complaint was under eye hollowness with 84% and followed by dark circles with 74%. It is seen that in over 40 year old patients, the most common complaint was dark circles with 80%. The filler was applied to 140 patients with cannula and to 38 patients with injection. The hyaluronic acid filler was implemented 0,83ml (0,4 1,25ml) total for each eye approximately. Patients were followed averagely 14 month (1 32 month). While the most frequent complication was ecchymosis in 14 patients (8%), the second one was edema which lasts over two weeks in 8 patients. Emboli was not seen in any patient. When the cannula and injection techniques were compared, a distinct difference was seen in terms of ecchymosis risk. The risk of ecchymosis was 3% with usage of cannula, whereas with needle, it was 26%. Besides, while ecchymosis caused by injection was seen in more extensive area, the area caused by cannula was more limited. As satisfaction evaluation, 93% patients graded the filler application as good. 5% patients thought that it was average and 2% of them scored it as bad. 4 patients who evaluated the application bad were the ones that their filler was applied with injection and occurred ecchymosis in their under eye. Result: The procedure of hyaluronic acid filler is an alternative treatment in the patients who have dark circles and hollows under eye. The procedure with cannula technique is safer for the patience satisfaction and risks of application, but this technique should be used with experienced doctor.

Improvement and optimization of the classical gastric biopsy culture technique for Helicobacter pylori diagnosis using trypsin.

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.

Comparison of Fluorescence Microscopy and Different Growth Media Culture Methods for Acanthamoeba Keratitis Diagnosis

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.