Variations in pre-analytical FFPE sample processing and bioinformatics: challenges for next generation molecular diagnostic testing in clinical pathology

Advances in cellular pathology techniques will improve diagnostic medicine. However, such improvements have to overcome many challenges including variations in pre-analytical sample processing, bioinformatics data analysis and clinical interpretation of data. In order to resolve such challenges, bioinformatics needs to become more tightly coupled to the experimental methodology development

Complete Sequence of pSAM7, an IncX4 Plasmid Carrying a Novel bla[sub]CTX-M-14b Transposition Unit Isolated from ' Escherichia coli ' and ' Enterobacter cloacae ' from cattle

The same plasmid carrying blaCTX-M-14b was identified from an Escherichia coli isolate and an Enterobacter cloacae isolate collected from cattle in the United Kingdom by complete plasmid sequencing. This 35,341-bp plasmid, pSAM7, had an IncX4 backbone that is 99% identical to that of pJIE143 from a human isolate in Australia. PCR screening identified pSAM7-like plasmids in three other E. coli isolates of different multilocus sequence types isolated from cattle on different farms in the United Kingdom

Determination of Urea Content in Dairy Cattle Feed by High Performance Liquid Chromatography via Refractive Index Detector: Method Optimization, Validation and Comparison with Spectro-Colorimetric Method

The aim of this study was to analyze the urea content of Dairy Cattle Feed (DCF) using High Performance Liquid Chromatography via Refractive Index Detector (HPLC-RID), without derivatization, along with spectro-colorimetric method that is the official control method of European Union (EU). Towards that goal, the analysis procedure for the method was established, method optimization and validation were carried out by spiking urea at 2% level to DCF, which is the upper limit allowed for use in ruminant feeds in Türkiye. In method validation; R2: 0.9997 coefficient of determination at 25-100000 mg/kg linear range, 75 mg/kg limit of detection (LOD) and 250 mg/kg limit of quantification (LOQ), 1.08% repeatability RSD, 1.84% reproducibility RSD and 95.10% recovery were obtained. The applicability of the method was proven at 25-100000 mg/kg linear range with real samples by spiking 2% urea in Urea-Free Dairy Cattle Feed (UF-DCF), Low Urea Dairy Cattle Feed (LU-DCF), and High Urea Dairy Cattle Feed (HU-DCF). The presence of urea in UF-DCF was detected with the spectro-colorimetric (at 420 and 435 nm wavelength) method, while urea in UF-DCF was not detected by the in-house HPLC-RID method, (P<0.05). The amount of urea in LU-DCF without spiking urea was detected by the same level as HPLC-RID method and the SC-435 method (P>0.05). For HU-DCF blank samples, higher urea amounts were detected with the in-house HPLC-RID method as compared to the spectro-colorimetric methods used in the study (P<0.05). In DCF spiked with 2.0% urea, the urea amount found in the in-house HPLC-RID method were higher than that of the spectro-colorimetric method (P<0.05). As a result, the in-house HPLC-RID method that is developed in this study has shown great promise to be a potential, applicable and valid method for determining the urea amount in DCF

Influence of the Isothermal Transformation Temperature on the Structure and Properties of Low-Carbon Steel

Purpose. The study is aimed at evaluating the effect of the isothermal transformation temperature on the structure and properties of low-carbon steel. Methodology. The material for the study was a 3 mm diameter wire made of mild steel with the following chemical composition: 0.21% C, 0.47% Mn, 1.2% Si, 0.1% Cr, 0.03% S, 0.012% P. The 0.3 m long wire samples were subjected to austenitizing at 920 °C for 8...9 min, after which they were held isothermally for 11 min at temperatures of 650...200 °C, followed by cooling in air. The strength, plastic properties, and strain hardening coefficient were determined from the analysis of tensile curves. Findings. It was found that a decrease in the temperature of isothermal transformation, starting from 450...400 °C, increases the amount of Widmannstätten ferrite due to the disappearance of polyhedral ferrite grains. At the same time, the number of areas with locally located dispersed cementite particles similar to pearlite colonies increases, and bainite crystals appear. Against the background of a sharp decrease in the strain hardening coefficient in the range of 450...400 °C, the ability of the bainite phase to undergo plastic deformation should be considered one of the reasons for the delay in density reduction. Originality. The effect of steel hardening with a decrease in the pearlite transformation temperature is based on the grinding of ferrite grains, an increase in the amount of Widmannstätten ferrite, and the dispersion of pearlite colonies. The strengthening effect of steel with a bainite structure is based on an increase in the degree of supersaturation of the solid solution with carbon atoms and dispersion hardening by particles of the carbide phase. Practical value. The optimal structural state of steel intended for the manufacture of such critical elements as a support beam, railroad car bogie, etc. is a mixture of phase components with different dispersion and morphology, and their quantitative ratio is determined by the operating conditions of a particular product

Analysis of the effects of inhibitor and activator systems (smad's proteins) of TGF-βs on chick neural tube closure; [Tavuk Nöral Tüp Kapanmasinda TGF-βs Inhibitör ve Aktivatör Sisteminin (Smad's Proteinleri) Etkilerinin Araştirilmasi]

The families of TGF-βs and Smads proteins that controls its intracellular signaling pathways are known to play a role in early neurulation. The aim of this study is to demonstrate distribution of TGF-βs (1, 2, 3) and Smads (1/2/3, 6, 7) proteins as a system in different hours of neural tube development of chick embryos. The SPF eggs were incubated at 37.8±2°C for 24th, 30th, 48th, 72nd h. After that, embryos were examined using immunohistochemistry and western blotting techniques. To the results, TGF-βs immunoreactivities (particularly TGF-β3) at the 24th, 30th and 48th h of chick development (during neural tube closure) were determined and decreased at the 72nd h (after neural tube closure), but expressions of TGF-βs were detected in all stage of embryos in western blotting. While Smad 1/2/3 immunoreactivitiy and expression was less than that of the Smad 6 and 7 at the 24th, it was increased at the 30th h. Smads proteins immunoreactivities were decresead at the 72nd h. In conclusion, the members of TGF-βs are play a role in chick neural tube closure, the secretions of TGF-βs are controlled different Smad proteins. In addition, immunoblotting results showed that TGF-βs and Smads proteins were effective in the development of all tissues and organs of the embryos

Presence of Hysterothylacium gadi aduncum (Rudolphi, 1802) (Anisakidae) in cultured Rainbow trout (Oncorhynchus mykiss Walbaum, 1792) in fresh water farms from Turkey and its mortality

The presence of the anisakid nematode Hysterothylacium gadi aduncum (Rudolphi, 1802) is reported for the first time in cultured rainbow trout (Oncorhynchus mykiss Walbaum, 1792) as a new host in freshwater farms from Turkey. This marine parasitic nematode is thought to complete the life cycle in fresh water and rainbow trout could act as the definitive host for this parasite under fresh water culture conditions. Mortalities in the rainbow trout infected with H. gadi aduncum (Rudolphi, 1802), were seen after three to four months period of feeding program with marine fish offal (Sprattus sprattus, Engraulis encrasicolus) and pellet fish food. In the present study, the main cause of mortalities was detected as stomach obstruction formed by aggregation of the adult nematodes which developed from infective third stage larvae. Original measurements and figures are presented.Publishe