Tenacibaculum maritimum CRISPR loci analysis and evaluation of isolate spoligotyping.

AIMS: We performed in silico analysis of CRISPRcas loci from Tenacibaculum maritimum, evaluated spoligotyping as a subtyping method and genotyped uncharacterized Turkish isolates from European sea bass by multilocus sequence typing (MLST). METHODS AND RESULTS: Spoligotyping was performed with primers designed to allow amplification and sequencing of whole CRISPR-arrays from 23 T. maritimum isolates. Twenty-three completed/draft genomes were also downloaded from the NCBI database and analysed. MLST of Turkish isolates was achieved with a well-established 7-gene scheme. Tenacibaculum maritimum genomes carry a structurally complete but partially defective class II CRISPRcas locus due to known amino acid substitutions in encoded Cas9 proteins. Our spacer identification suggests that the host range of bacteriophage P2559Y and Vibrio phage nt-1 include T. maritimum and that the most recurrent infection recorded by isolates has been with Tenacibaculum phage PTm5. Thirty-eight isolates with this CRISPRcas locus belonged to 25 spoligotypes and to 24 sequence types by MLST, respectively. According to MLST, T. maritimum isolates from Turkey are most related to previously defined sequence types ST3, ST40 and ST41 isolates from Spain, Malta and France. CONCLUSIONS: The evaluated spoligotyping offers discriminatory power comparable to MLST. SIGNIFICANCE AND IMPACT OF THE STUDY: Spoligotyping has potential as a quick, easy and cheap tool for subtyping of T. maritimum isolates.

Mechanic ileus due to retroperitoneal migration of total hip prosthesis; A case report.

Intrapelvic migration of total hip prosthesis is a rare but severe complication of total hip arthroplasty that can cause severe outcomes for elderly patients. A 78-year-old female patient was referred to our hospital with the complaint of no gas-stool excretion for 3-4 days, abdominal distension, nausea, vomiting, and a preliminary diagnosis of ileus. Computed tomography showed the migration of the left total hip prosthesis to the pelvis, causing a hematoma around the prosthesis and mechanical ileus due to the compression of the hematoma. To our knowledge, this case report is the only reported mechanic ileus due to migration of total hip prosthesis. Although postop paralytic ileus is one of the complications of total hip arthroplasty, mechanical ileus has not been described before. This case report shows that mechanical ileus might be an unreported complication of total hip arthroplasty. It should be kept in mind that mechanical ileus complications may also occur after hip arthroplasty.

Environmental biofilm communities associated with early-stage common dentex (Dentex dentex) culture.

AIMS: To describe the biofilm microbiota associated with various feeding phases during larval common dentex (Dentex dentex) culture. METHODS AND RESULTS: A targeted metagenomic (metagenetic) study was performed by means of 16S rRNA gene-based PCR and NextGen pyrosequencing. The resulting dataset was scrutinized with microbial community analysis software (r packages) using r/Rstudio. While median observed and estimated alpha-diversities were 171 ± 38 and 207 ± 27 taxa, respectively, 72·1-85·8% of individual biofilm communities comprised only 27-46 taxa. Members of the genus Methylobacterium and family Rhodobacteraceae dominated biofilms formed during all feeding phases while genera Nannochloropsis and Tetraselmis microalgae were major constituents of biofilms during rotifer live feeding. Both potential fish pathogenic genera, for example, Vibrio and putatively probiotic taxa, for example, Phaeobacter gallaeciensis were identified. CONCLUSIONS: Relatively stable biofilm communities were identified during each feeding phase but varied significantly between feeding phases, most likely in response to the introduction of live feed/microalgae-associated bacteria into rearing tanks. SIGNIFICANCE AND IMPACT OF THE STUDY: The structure of the bacterial communities identified represents a 'template' for successful larval dentex culture and provides a foundation for future investigations into failed production cycles.

Human eyelash characterization.

BACKGROUND: Few biological data on human eyelash follicles have been reported in the literature. OBJECTIVES: To characterize eyelash follicle growth, cycle and morphology, and further investigate the biological mechanisms that determine eyelash length, curl and pigmentation, compared with scalp hair follicle. METHODS: Twenty-nine caucasian female volunteers aged between 26 and 60 years were enrolled in the study to provide eyelashes. Four of these volunteers were followed weekly for 9 months to characterize their eyelash cycle. Eyelash length and time of renewal were measured using a high-resolution camera and image analysis. Immunohistological study of the bulbs were performed on eyelid biopsies from 17 patients requiring block excision for ectropion repair. RESULTS: The calculated durations of anagen phase and complete cycle of the eyelashes were 34 + or - 9 and 90 + or - 5 days, respectively. Eyelash follicle growth rate was quite variable, with an average rate of 0.12 + or - 0.05 mm daily. Eyelash follicle morphology was very close to that of the scalp hair follicle, but some remarkable differences were noticed. For example, the K19-positive epithelial stem cell population was spread all along the follicle and not split into two reservoirs as seen in scalp hair follicles. Some asymmetry was detected in HSPG and CSPG, as well as K38 (formerly Ha8) and K82 (formerly Hb2) distribution, similar to that observed in curly hair. Finally, dopachrome tautomerase was found expressed in eyelash follicle melanocytes, while it was strikingly absent in scalp hair follicle melanocytes. CONCLUSIONS: The eyelash is structurally very close to curly hair but some biological processes related to follicle cycle and pigmentation differ markedly.

Real time PCR detection of Piscirickettsia salmonis from formalin-fixed paraffin-embedded tissues.

Piscirickettsia salmonis is the causative agent of piscirickettsiosis, a transmissible disease of salmonid fish. Diagnosis of piscirickettsiosis has traditionally been based upon identification of typical pathological changes by histological investigation, with confirmation by immunohistochemistry on formalin-fixed, paraffin-embedded tissues. However, implementation of more rapid confirmatory techniques, preferably with higher levels of sensitivity and possibilities for quantification, is desirable. A real-time polymerase chain reaction (PCR) assay was designed for specific detection of P. salmonis and tested on samples extracted from formalin-fixed paraffin-embedded material. Construction of a PCR-target mimic allowed determination of detection limits, linearity of the real-time PCR and quantitative detection of P. salmonis. The present study demonstrates the capability of the described real time PCR assay for detection of P. salmonis from paraffin-embedded material with a high degree of sensitivity and specificity. Implementation of this assay constitutes an important development for a rapid and secure diagnosis of piscirickettsiosis.

Evaluation of leptin, adiponectin, and ghrelin levels in patients with acne vulgaris.

BACKGROUND: The research evaluating adipokines are very few in patients with acne vulgaris. The hypothesis that hyperinsulinemic and high glycemic index diet plays a role in the pathogenesis of acne is still controversial. In this study, we aimed to evaluate adipokines such as leptin (L), adiponectin (A), ghrelin and A levels, and A/L rates that indicate insulin resistance in nonobese patients with severe acne vulgaris. MATERIAL AND METHOD: Thirty patients who are nonobese with moderate acne vulgaris, aged 18 to 25 years, and 15 age-sex compatible controls were included in our study. The acne lesions were assessed using the Global Acne Grading Scale (GAGS). All participants were evaluated for the parameters that may affect the metabolism of serum L, A, and ghrelin levels in blood, and their body mass index were calculated. The significance level was determined as p ≤ 0.05. RESULTS: Of the 30 patients, 17 were women and 13 were men. The mean age was 20.60 years and the mean duration of the disease were 2.8 years. All of patients had moderate acne vulgaris (GAGS 19-30). Of the 15 controls, 11 were women and 4 were men. The mean age was 21.20 years. There were not a statistically significant difference in L, ghrelin, A levels, and A/L ratio between the two groups. CONCLUSIONS: Adipokines may have a role in the pathogenesis of acne vulgaris. L, A, ghrelin, and insulin resistance may not participate in the responsible mechanisms in nonobese patients with moderate acne vulgaris.