COMMON NONCONFORMITIES DURING PROCEDURE FOR ACCREDITATION OF THE FOOD TESTING LABORATORIES IN THE R. MACEDONIA

During laboratory assessment in the frame of the accreditation procedure, they faced with number of nonconformities which are challenge for receiving the Certificate for accreditation and appropriate corrective measures shall be undertaken. Requirements that should be met by the food testing laboratories in the Republic of Macedonia, in order to gain accreditation certificate, contained in the standard МКС ENISO/IEC 17025:2006, ILAC (International Laboratory Accreditation Cooperation) document, ЕА (European Cooperation for Accreditation) document, Regulations and Procedures of the Institute for Accreditation of the Republic of Macedonia. In this paper the analysis is made and different proposals are given for the different ways of fulfilling of those requirements. The aim of this study is to emphasis the common nonconformities which are observed during realization of the Procedure of Accreditation of food testing laboratories and corrective measures undertaken. This investigation is of huge meaning for food testing laboratories which are already accredited and those which are planning to be accredited in the Republic of Macedonia. Furthermore, it is a very important for the Institute for accreditation of the Republic of Macedonia to come to conclusions for the weakest sides of the food testing laboratories and the assessment thereof. Such conclusions should initiate undertaking appropriate measures for improvement the Institute’s lead and technical assessors work towards nonconformities interpretation, identification and acceptance of the most suitable corrective measures

DIFFERENT APPROACHES IN ANALYZING CHYMOSIN PURITY

Chymosin is a specific proteolytic enzyme found in rennet, and is the key enzyme in cheese production classified in the aspartic endopeptidases (EC 3.4.23.4). The aim of this study was to determine the purity of different commercially available chymosins and its equivalents using electrophoretic and chromatographic techniques. Chymosins produced by the company Chr. Hansen, CHY-MAX 200 and CHY-MAX Plus, CHY-MAX PowderExtra NB, as well as Maxiren 1800 Granulate from the company DSM, Sirnik from SZR – Travnik, Kraljevo and Planika from Mikroprocessing, Bileca were used as materials for this study. The purity level of the commercially available enzymes was analyzed using electrophoretic (sodium dodecyl sulfate polyacrylamide gel electrophoresis or SDS-PAGE) and chromatographic (Rapid Resolution Liquid Chromatography or RRLC) techniques. Results showed no presence of undeclared protein fractions due to inappropriate purification process in the samples except for CHY-MAX М 200 which had two protein fractions, most likely as a result of a polymorphism. All the CHY-MAX and Maxiren samples have chymosin as the active component (36 kDa), except for Planika and Sirnik which have a natural protease from R. miehei. Chromatographic analysis showed that beside the active component (chymosin), the preservative sodium benzoate was present in varying concentrations in all but CHY-MAX PowderExtra NB

Homologous Seminal Plasma and Glutathione Promote Pre-capacitation Motility and Structural Stability of Cryopreserved Ram Spermatozoa

Reduced glutathione (GSH) and homologous ram seminal plasma (HSP), used as additives in cryopreserving (CP) media prior to freezing, showed conflicting results in retaining structural integrity and progressive motility in post-thawed ram spermatozoa. The aims of this research were (1) to assess the effect of GSH and/or HSP supplementation via soybean-lecithin CP extender on cryopreserved ram spermatozoa viability, morphology and motility pattern; and (2) to assess the effect of incubation in the context of the previous aim. Quantitatively and qualitatively, homogenized and pooled ram ejaculates (N=10) were extended with one of the following extenders Control (C)-tris-based, GSH and HSP-free, experimental-1 (E1)-C + GSH 5 mM, experimental-2 (E2)-C + HSP 20 % and experimental-3 (E3)-GSH 5 mM + HSP 20 %. Following thawing, samples were taken at 0-A nd 3-hours from each group (n=10) and were assessed for spermatozoa viability, morphology, and motility pattern. C-0h samples yielded a spermatozoa population with low viability, altered head morphology and highly deviated motility pattern. E3-3h samples yielded spermatozoa with unaffected viability, head morphology and high progressive motility. In conclusion, E3 extender added to cryopreserved-thawed ram spermatozoa is most efficient in obtaining high viability, unaltered head morphology, and progressive motility

Bcl-2 and β1-integrin predict survival in a tissue microarray of small cell lung cancer.

INTRODUCTION: Survival in small cell lung cancer (SCLC) is limited by the development of chemoresistance. Factors associated with chemoresistance in vitro have been difficult to validate in vivo. Both Bcl-2 and β(1)-integrin have been identified as in vitro chemoresistance factors in SCLC but their importance in patients remains uncertain. Tissue microarrays (TMAs) are useful to validate biomarkers but no large TMA exists for SCLC. We designed an SCLC TMA to study potential biomarkers of prognosis and then used it to clarify the role of both Bcl-2 and β(1)-integrin in SCLC. METHODS: A TMA was constructed consisting of 184 cases of SCLC and stained for expression of Bcl-2 and β(1)-integrin. The slides were scored and the role of the proteins in survival was determined using Cox regression analysis. A meta-analysis of the role of Bcl-2 expression in SCLC prognosis was performed based on published results. RESULTS: Both proteins were expressed at high levels in the SCLC cases. For Bcl-2 (n=140), the hazard ratio for death if the staining was weak in intensity was 0.55 (0.33-0.94, P=0.03) and for β(1)-integrin (n=151) was 0.60 (0.39-0.92, P=0.02). The meta-analysis showed an overall hazard ratio for low expression of Bcl-2 of 0.91(0.74-1.09). CONCLUSIONS: Both Bcl-2 and β(1)-integrin are independent prognostic factors in SCLC in this cohort although further validation is required to confirm their importance. A TMA of SCLC cases is feasible but challenging and an important tool for biomarker validation

A global analysis of Y-chromosomal haplotype diversity for 23 STR loci

In a worldwide collaborative effort, 19,630 Y-chromosomes were sampled from 129 different populations in 51 countries. These chromosomes were typed for 23 short-tandem repeat (STR) loci (DYS19, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS385ab, DYS437, DYS438, DYS439, DYS448, DYS456, DYS458, DYS635, GATAH4, DYS481, DYS533, DYS549, DYS570, DYS576, and DYS643) and using the PowerPlex Y23 System (PPY23, Promega Corporation, Madison, WI). Locus-specific allelic spectra of these markers were determined and a consistently high level of allelic diversity was observed. A considerable number of null, duplicate and off-ladder alleles were revealed. Standard single-locus and haplotype-based parameters were calculated and compared between subsets of Y-STR markers established for forensic casework. The PPY23 marker set provides substantially stronger discriminatory power than other available kits but at the same time reveals the same general patterns of population structure as other marker sets. A strong correlation was observed between the number of Y-STRs included in a marker set and some of the forensic parameters under study. Interestingly a weak but consistent trend toward smaller genetic distances resulting from larger numbers of markers became apparent

Effect of Intraoperative High Positive End-Expiratory Pressure (PEEP) with Recruitment Maneuvers vs Low PEEP on Postoperative Pulmonary Complications in Obese Patients: A Randomized Clinical Trial

Importance: An intraoperative higher level of positive end-expiratory positive pressure (PEEP) with alveolar recruitment maneuvers improves respiratory function in obese patients undergoing surgery, but the effect on clinical outcomes is uncertain. Objective: To determine whether a higher level of PEEP with alveolar recruitment maneuvers decreases postoperative pulmonary complications in obese patients undergoing surgery compared with a lower level of PEEP. Design, Setting, and Participants: Randomized clinical trial of 2013 adults with body mass indices of 35 or greater and substantial risk for postoperative pulmonary complications who were undergoing noncardiac, nonneurological surgery under general anesthesia. The trial was conducted at 77 sites in 23 countries from July 2014-February 2018; final follow-up: May 2018. Interventions: Patients were randomized to the high level of PEEP group (n = 989), consisting of a PEEP level of 12 cm H2O with alveolar recruitment maneuvers (a stepwise increase of tidal volume and eventually PEEP) or to the low level of PEEP group (n = 987), consisting of a PEEP level of 4 cm H2O. All patients received volume-controlled ventilation with a tidal volume of 7 mL/kg of predicted body weight. Main Outcomes and Measures: The primary outcome was a composite of pulmonary complications within the first 5 postoperative days, including respiratory failure, acute respiratory distress syndrome, bronchospasm, new pulmonary infiltrates, pulmonary infection, aspiration pneumonitis, pleural effusion, atelectasis, cardiopulmonary edema, and pneumothorax. Among the 9 prespecified secondary outcomes, 3 were intraoperative complications, including hypoxemia (oxygen desaturation with Spo2 ≤92% for >1 minute). Results: Among 2013 adults who were randomized, 1976 (98.2%) completed the trial (mean age, 48.8 years; 1381 [69.9%] women; 1778 [90.1%] underwent abdominal operations). In the intention-to-treat analysis, the primary outcome occurred in 211 of 989 patients (21.3%) in the high level of PEEP group compared with 233 of 987 patients (23.6%) in the low level of PEEP group (difference, -2.3% [95% CI, -5.9% to 1.4%]; risk ratio, 0.93 [95% CI, 0.83 to 1.04]; P =.23). Among the 9 prespecified secondary outcomes, 6 were not significantly different between the high and low level of PEEP groups, and 3 were significantly different, including fewer patients with hypoxemia (5.0% in the high level of PEEP group vs 13.6% in the low level of PEEP group; difference, -8.6% [95% CI, -11.1% to 6.1%]; P <.001). Conclusions and Relevance: Among obese patients undergoing surgery under general anesthesia, an intraoperative mechanical ventilation strategy with a higher level of PEEP and alveolar recruitment maneuvers, compared with a strategy with a lower level of PEEP, did not reduce postoperative pulmonary complications. Trial Registration: ClinicalTrials.gov Identifier: NCT02148692

Effect of Intraoperative High Positive End-Expiratory Pressure (PEEP) With Recruitment Maneuvers vs Low PEEP on Postoperative Pulmonary Complications in Obese Patients: A Randomized Clinical Trial.

IMPORTANCE An intraoperative higher level of positive end-expiratory positive pressure (PEEP) with alveolar recruitment maneuvers improves respiratory function in obese patients undergoing surgery, but the effect on clinical outcomes is uncertain