Ovine PAPPA2 gene coding variants are linked to decreased fecal egg shedding in native Turkish sheep naturally infected with gastrointestinal nematodes.

In this study, the association between PAPPA2 coding variants and gastrointestinal (GI) nematode fecal egg count (FEC) score in adult Turkish sheep was investigated. For this purpose, the FEC score was determined in adult sheep from six breeds: Karacabey Merino (n = 137), Kivircik (n = 116), Cine capari (n = 109), Karakacan (n = 102), Imroz (n = 73), and Chios (n = 50). Sheep were classified as shedders or non-shedders within breeds and flocks. The first group was the fecal egg shedders (> 50 per gram of feces), and the second group was the no fecal egg shedders (≤ 50 per gram of feces). The exon 1, exon 2, exon 5, exon 7, and a part of 5'UTR of the ovine PAPPA2 gene were genotyped by Sanger sequencing of these two groups. Fourteen synonymous and three non-synonymous single-nucleotide polymorphisms (SNPs) were found. The non-synonymous SNPs, D109N, D391H, and L409R variants, are reported for the first time. Two haplotype blocks were constructed on exon 2 and exon 7. The specific haplotype, C391G424G449T473C515A542 on the exon 2 that carries the 391H variant, was tested against four other common haplotypes. Our results indicate that C391G424G449T473C515A542 haplotype was significantly associated with fecal egg shedding status in adult Turkish sheep (p-value, 0.044).

The tetraspanin Tspan15 is an essential subunit of an ADAM10 scissor complex.

A disintegrin and metalloprotease 10 (ADAM10) is a transmembrane protein essential for embryonic development, and its dysregulation underlies disorders such as cancer, Alzheimer's disease, and inflammation. ADAM10 is a "molecular scissor" that proteolytically cleaves the extracellular region from >100 substrates, including Notch, amyloid precursor protein, cadherins, growth factors, and chemokines. ADAM10 has been recently proposed to function as six distinct scissors with different substrates, depending on its association with one of six regulatory tetraspanins, termed TspanC8s. However, it remains unclear to what degree ADAM10 function critically depends on a TspanC8 partner, and a lack of monoclonal antibodies specific for most TspanC8s has hindered investigation of this question. To address this knowledge gap, here we designed an immunogen to generate the first monoclonal antibodies targeting Tspan15, a model TspanC8. The immunogen was created in an ADAM10-knockout mouse cell line stably overexpressing human Tspan15, because we hypothesized that expression in this cell line would expose epitopes that are normally blocked by ADAM10. Following immunization of mice, this immunogen strategy generated four Tspan15 antibodies. Using these antibodies, we show that endogenous Tspan15 and ADAM10 co-localize on the cell surface, that ADAM10 is the principal Tspan15-interacting protein, that endogenous Tspan15 expression requires ADAM10 in cell lines and primary cells, and that a synthetic ADAM10/Tspan15 fusion protein is a functional scissor. Furthermore, two of the four antibodies impaired ADAM10/Tspan15 activity. These findings suggest that Tspan15 directly interacts with ADAM10 in a functional scissor complex.

The use of preanalytical quality indicators: a Turkish preliminary survey study.

OBJECTIVES: The utilization of reliable quality indicators (QIs) proven to be suitable for monitoring and improvement tools is one of the best choices to minimize of the risk of errors in all laboratory processes called as total testing process (TTP). In 2008, a Working Group "Laboratory Errors and Patient Safety" (WG-LEPS) established by International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) developed the Model of Quality Indicators (MQI) complying with requirements of the ISO 15189:2012 standard for laboratory accreditation. They have also been dealing with harmonizing the QIs in most laboratories worldwide since then. The present study was set out to investigate the frequency of using IFCC WG-LEPS' pre-QIs by Turkish laboratories and to assess the conformity of them, by taking into account Turkey's conditions. METHODS: A survey consisting nine questions was applied in 81 laboratories using SurveyMonkey. RESULTS: According to the survey results, most of the laboratories reported they have used pre-QIs in the quality standards of health prepared by Turkish Ministry of Health (MOH). A part of IFCC WG-LEPS' pre-QIs were being utilized by more than 80% of the laboratories, the rest of which only used by 10% of laboratories. CONCLUSIONS: The majority of the medical laboratories have been using the pre-QIs included in the guidelines of Quality Standards prepared by the MOH. The pre-QIs are partially compatible with IFCC WG-LEPS' pre-QIs. The definitions of IFCC WG-LEPS' pre-QIs may also be revised to make them more clear and understandable by IFCC WG-LEPS. The insufficiency of Health Information Management Systems (HIMS) limits the use of pre-QIs proposed by IFCC WG-LEPS. Finally, the education of relevant personnel about the use of HIMS and pre-QIs is very crucial to harmonize and to extend the use of IFCC WG-LEPS' pre-QIs in Turkish medical biochemistry laboratories.

Documentation of extensive genetic diversity in the Ovar-DRB1 gene in native Turkish sheep.

Indigenous breeds have a high level of genetic diversity that might contribute to develop animal breeds with desired traits such as disease resistance and high productivity. Major histocompatibility complex (MHC) is a key component of adaptive immune system and consists of highly polymorphic genes that take part in adaptive immune response and disease resistance. Exploring and understanding the effect of polymorphisms in MHC could be beneficial to future animal breeding strategies. In this study, we sequenced the highly polymorphic Exon2 of the ovine DRB1 gene using Sanger sequencing to explore the diversity of this gene in six indigenous Turkish sheep breeds and two crossbreeds. In total, 894 haplotypes from 447 sheep were investigated, and 69 different haplotypes including 27 novel ones were identified. Among the identified haplotypes there were common and breed specific haplotypes. There was a relatively high diversity of the alleles within indigenous breeds. Allelic diversity patterns were mostly associated with geographical differences. The results of this study highlight the genetic variation within indigenous breeds which has important implications for biodiversity and the adaptability of breeds to specific environments. There is value to further studies which include other genomic regions and traits, and these could guide breeding strategies.

Association between ovine Toll-like receptor 4 (TLR4) gene coding variants and presence of Eimeria spp. in naturally infected adult Turkish native sheep.

Coccidiosis caused by Eimeria spp. is a protozoan disease prevalent in farm animals, and it is responsible for serious economic losses especially in young animals. It has been popular to breed disease-resistant animals due to the concern about food safety, animal welfare, and public health. Toll-like receptor (TLR) gene family plays a key role in the innate immune system participating in host-antigen interaction, therefore, they are candidate genes for breeding disease-resistant animals. In the present study, possible genetic associations between TLR4 gene coding variants and the presence of Eimeria spp. in adult Turkish sheep were investigated. For this purpose, the presence of Eimeria spp. in fecal samples from six native Turkish sheep were determined, and approximately 1450 bp region in the 3rd exon of the ovine TLR4 gene was sequenced. Ten nonsynonymous and four synonymous single nucleotide polymorphisms (SNPs) were detected in the targeted region. Statistical analyses revealed that the SNP at the codon at 356th position encoding Leucine instead of Phenylalanine (F356L) was significantly associated with the presence of Eimeria spp. It was found that the individuals carrying at least one Leucine amino acid sequence at this position have 2.3-fold more risk for the presence of Eimeria spp.

Evaluation of CD109, PCP4 and SEMA3D genes for their association with Ovine Johne's disease in Turkish sheep.

Johne's disease is a chronic, contagious, zoonotic disease that affects numerous species including livestock and sometimes humans. The disease is globally distributed in sheep populations and caused by Mycobacterium avium Subsp. paratuberculosis (MAP). A previous genome-wide association study identified single nucleotide polymorphism (SNP) markers associated with OJD serostatus in CD109, PCP4, and SEMA3D genes. Our aim was to evaluate the same markers for association with OJD seroprevalence in Turkish sheep in a retrospective matched case-control study. The serological status for OJD in 1801 sheep was determined for four native and four composite breeds from three research flocks. One hundred eleven matched case-control pairs were constructed according to breed type and age from 1750 comingled ewes reared in the same environment. A Single Nucleotide Primer Extension (SNuPE) assay was designed to genotype PCP4-Intron 1, PCP4-3'UTR, SEMA3D, CD109-intron 2 and CD109-intron 8 markers and a McNemar's test was performed on the matched pairs. An association with these five markers was not detected with the OJD serostatus in Turkish sheep (power of detection, 0.95; odds ratio >3; McNemar's p < .05). Thus, a wider search may be needed to identify any major underlying genetic risk factors for OJD in Turkish sheep.

Detecting fecal egg count (FEC) for gastrointestinal nematodes of adult Turkish sheep with different scrapie related PRNP haplotypes.

Scrapie is a transmissible spongiform encephalopathy caused by prions and leads to neurodegeneration in the Central Nervous System (CNS) of sheep and goats. Genetic resistance/susceptibility to scrapie is well studied and it is known that the variations of 136th, 154th and 171st codons at the ovine PRNP gene have a major effect on the development of the disease. Many studies demonstrated that selection for PRNP genotypes has not influenced other performance traits, nevertheless, there is a knowledge gap about the possible link between the PRNP gene and the status of the other important diseases that affect the sheep population worldwide. In the present study, we tested whether there is an association between scrapie-related PRNP genotypes and fecal egg count (FEC) of gastrointestinal nematodes in seven adult Turkish sheep breeds. For this purpose, FEC scores of studied sheep (n = 253) were determined and the same animals were genotyped for the PRNP gene. Finally, an association analysis was performed for scrapie resistant (ARR), susceptible (VRQ), and wild-type (ARQ) haplotypes. Based on our statistical analysis, it is concluded that PRNP genotypes have no positive or negative effect on the FEC scores of adult sheep.

Capsule endoscopy in a network cooperation: assessment of the experience in 822 patients.

INTRODUCTION: Capsule endoscopy (CE) is firmly established as a standard procedure in the diagnostic algorithm of occult or obscure gastrointestinal bleeding and Crohn's disease. Despite its excellent diagnostic yield, missing expertise, reading time and financial expenditure limit an area-wide availability. A multicentric cooperation might compensate these disadvantages. METHODS: CE device was bought by a central hospital (CH). Requested equipment is transported to the network partner (NP) and the procedure performed at the spot in personal responsibility. Video reading is exclusively done in the CH. RESULTS: Within 10 years, 822 CE (438 m., 384 f.; 63 ± 17 (13-92) years) were performed by 18 cooperating gastroenterological departments. 587/822 (71%) CE were done at NP, 235/822 (29%) in the CH. Between 2002 (n = 39) and 2011 (n = 123) the annual number of CE increased threefold. 95% of the capital investment in each cooperating hospital could be avoided by sharing one workstation within the network. Leading indication for CE was suspected mid-GI-bleeding (80%). Mean latencies between requested date and actual examination were 0 and between equipment's return and report 2 days. 45/191(24%) flexible enteroscopies performed in the CH followed CE findings from NP. DISCUSSION: Our 10 years experience show that mobile use of CE is feasible providing quality parameters similar to a single center solution, increases the number of CE investigations, therefore, improves reading expertise and enables both an area-wide and economic offer for this technique. Additionally, patients with the need for invasive enteroscopy are identified and attracted to that NP who provides an invasive SB endocopy device.

Evaluation of the clinical chemistry tests analytical performance with Sigma Metric by using different quality specifications - Comparison of analyser actual performance with manufacturer data.

INTRODUCTION: The interest in quality management tools/methodologies is gradually increasing to ensure quality and accurate results in line with international standards in clinical laboratories. Six Sigma stands apart from other methodologies with its total quality management system approach. However, the lack of standardization in tolerance limits restricts the advantages for the process. Our study aimed both to evaluate the applicability of analytical quality goals with Roche Cobas c 702 analyser and to determine achievable goals specific to the analyser used. MATERIALS AND METHODS: The study examined under two main headings as Sigmalaboratory and Sigmaanalyser. Sigmalaboratory was calculated using internal and external quality control data by using Roche Cobas c 702 analyser for 21 routine biochemistry parameters and, Sigmaanalyser calculation was based on the manufacturer data presented in the package inserts of the reagents used in our laboratory during the study. Sigma values were calculated with the six sigma formula. RESULTS: Considering the total number of targets achieved, Sigmaanalyser performed best by meeting all CLIA goals, while Sigmalaboratory showed the lowest performance relative to biological variation (BV) desirable goals. CONCLUSIONS: The balance between the applicability and analytical assurance of "goal-setting models" should be well established. Even if the package insert data provided by the manufacturer were used in our study, it was observed that almost a quarter of the evaluated analytes failed to achieve even "acceptable" level performance according to BV-based goals. Therefore, "state-of-the-art" goals for the Six Sigma methodology are considered to be more reasonable, achievable, and compatible with today's technologies.

A novel 2 bp deletion variant in Ovine-DRB1 gene is associated with increased Visna/maedi susceptibility in Turkish sheep.

Visna/maedi (VM) is a multisystemic lentivirus infection of sheep that affecting sheep industry across the globe. TMEM154 gene has been identified to be a major VM-associated host gene, nevertheless, a recent study showed that the frequency of the VM-resistant TMEM154 haplotypes was very low or absent in indigenous sheep. Thus, the present study was designed to determine other possible co-receptors associated with VM. For this purpose, DRB1 gene, which is renowned for its role in host immune response against various diseases was targeted. A total number of 151 case-control matched pairs were constructed from 2266 serologically tested sheep. A broad range of DRB1 haplotype diversity was detected by sequence-based genotyping. Moreover, a novel 2 bp deletion (del) in the DRB1 intron 1 was identified. For the final statistic, the sheep carrying VM-resistant TMEM154 diplotypes were removed and a McNemar's test with a matched pairs experimental design was conducted. Consequently, it was identified for the first time that the 2 bp del variant is a genetic risk factor for VM (p value 0.002; chi-square 8.31; odds ratio 2.9; statistical power 0.90) in the dominant model. Thus, negative selection for 2 bp del variant could decrease VM infection risk in Turkish sheep.

Association of TLR2 haplotypes encoding Q650 with reduced susceptibility to ovine Johne's disease in Turkish sheep.

Ovine Johne's disease (OJD) is caused by Mycobacterium avium subsp. paratuberculosis (MAP) and carries a potential zoonotic risk for humans. Selective breeding strategies for reduced OJD susceptibility would be welcome tools in disease eradication efforts, if available. The Toll-like receptor 2 gene (TLR2) plays an important signaling role in immune response to MAP, and missense variants are associated with mycobacterial infections in mammals. Our aim was to identify and evaluate ovine TLR2 missense variants for association with OJD in Turkish sheep. Eleven TLR2 missense variants and 17 haplotype configurations were identified in genomic sequences of 221 sheep from 61 globally-distributed breeds. The five most frequent haplotypes were tested for OJD association in 102 matched pairs of infected and uninfected ewes identified in 2257 Turkish sheep. Ewes with one or two copies of TLR2 haplotypes encoding glutamine (Q) at position 650 (Q650) in the Tir domain were 6.6-fold more likely to be uninfected compared to ewes with arginine (R650) at that position (CI95 = 2.6 to 16.9, p-value = 3.7 × 10-6). The protective TLR2 Q650 allele was present in at least 25% of breeds tested and thus may facilitate selective breeding for sheep with reduced susceptibility to OJD.

[Completion pneumonectomy: indications, complications, and results]. / Tamamlayici pnömonektomiler: endikasyonlari, komplikasyonlari ve sonuçlari.

Completion pneumonectomy is reported to be associated with high morbidity and mortality, especially when performed in patients with benign diseases. In our study we aimed to evaluate all patients underwent completion pneumonectomy in our clinic and to compare indications, complications and postoperative results with the literatures. Between January 1987 and December 2001, 27 consecutive patients who underwent completion pneumonectomy in our clinic were retrospectively reviewed. Postoperative morbidity and mortality rates were calculated according to indications and the results were compared to the standard pneumonectomies. There were 27 patients, 13 (48.1%) women and 14 (51.9%) men, with a median age of 26 (range, 10 to 62 years). Completion pneumonectomy was performed for benign diseases in 23 (85.2%) patients and for malign diseases in 4 (14.8%). Malign indications included 2 second primary tumors and 2 local recurrences. In the group with benign diseases; completion pneumonectomy was performed for tuberculosis in 5, bronchiectasis in 14, bronchopleural fistula in 2 and necrosis of lung in 2. Hospital mortality was 7.4% including 1 intraoperative and 1 postoperative deaths and both of them had undergone completion pneumonectomy for benign diseases. Complications occurred in 9 (33.3 %) patients, bronchopleural fistula + empyema were seen in 6 patients, cardiac rhythm disorders in 2 and wound infection in 1. All complications occurred in the patients operated for benign indications (39.1%). Completion pneumonectomy can be performed with an acceptable morbidity and mortality (similar to standard pneumonectomy) in selected cases. But the complication risk is higher in benign diseases, especially in tuberculosis. Surgical technique is important to avoid serious complications such as bronchopleural fistula and empyema.

Clinicopathologic demonstration of complex bronchopulmonary foregut malformation.

Bronchopulmonary foregut malformations are rare and often prove to be extremely difficult to diagnose. The presented case is a 44-year-old man complaining about cough and hemoptysis. These symptoms continued despite medical treatment. A computed thorax tomographic scan depicted a paravertebral cystic lesion. The patient underwent surgical exploration, and communication between the cyst and the esophagus was found. A left lower lobectomy was performed. The pathologic result confirmed a duplication cyst of mixed bronchogenic and esophageal type with bronchiectasis of the lower lobe. We present this unusual malformation complex especially in an adult with the review of the literature.