Abstract Objective: Tinnitus can result from auditory system reorganization due to neural activity dysfunctions. Auditory stimulation can cause temporary or persistent tinnitus alleviation by altering the neural generators. The present study investigated the changes in Late-Latency Auditory Evoked Potentials (LLAEPs) after tinnitus suppression using auditory stimulation with short-term Residual Inhibition (RI) and long-term Tinnitus Masker (TM). Methods: The study included 40 participates with equal numbers in the Tinnitus Group (TG), including the chronic tinnitus subjects, and the Control Group (CG), including matched volunteers. The participants had normal hearing in conventional audiometry. All the participants underwent LLAEP recording pre-intervention and after a one-minute auditory stimulation (RI), as well as a pre-intervention Dichotic Digit Test (DDT) as a behavioral assessment of the selective attention. Moreover, TG underwent a 3-month course of TM, a third LLAEP recording post-TM, a second DDT post-TM, and the Tinnitus Handicap Inventory (THI) before and after the TM. Results: In the baseline LLAEP recording, the TG had significantly later N1, P3a, and P3b latencies, as well as significantly lower P3a and P3b amplitudes. The second LLAEP recording showed a significant intragroup increase in P3a amplitude and a significant decrease in P3a latency in the TG, while no significant intragroup difference was observed in the CG. In the third LLAEP recording performed on TG, the P3a amplitude and latency had significant changes compared to the second recording, while the N1 latency was significantly decreased. Moreover, the DDT and THI scores had significant improvement after the TM in TG. Conclusions: The present study showed the neurophysiological changes after short-term (RI) and long-term (TM) auditory stimulations in tinnitus subjects. The LLAEP changes suggest that these interventions lead to tinnitus suppression through common mechanisms. The electrophysiological observation was also confirmed using behavioral assessments. Level of evidence: This study type is a “comparative study” with the level of evidence “3”.

SUMMARY OBJECTIVE: Polycystic ovary syndrome is a hormonal disorder that normally affects women of reproductive age in the range of 18–44 years. This study aimed to investigate the allelic frequencies of two polymorphisms, IRS rs18012781 and INSR rs1799817, which are suspected to be involved in polycystic ovary syndrome. METHODS: The samples were obtained from the patients admitted to the Near East University Hospital, Department of Gynecology and Obstetrics. The samples were divided into two groups: control and polycystic ovary syndrome groups. Blood samples were collected from 55 women in the control group and 65 samples from the patient group. DNA from whole blood was obtained. The allelic frequencies of single-nucleotide polymorphisms were determined using real-time PCR. Results were presented as the heterozygous and homozygous state of the single-nucleotide polymorphisms. RESULTS: There were no significant differences in the allelic frequencies of the single-nucleotide polymorphisms between the patient and control groups. Further statistical analysis investigating the INSR Tm using the Mann-Whitney U test value revealed that there was no difference in the homozygous and heterozygous state of INSR rs1799817. The result of this study showed that there was no statistically significant difference between the allelic frequencies of IRS1 rs1801278 and INSR rs1799817 between the patient and control groups. CONCLUSION: These single-nucleotide polymorphisms do not seem to modify the risk of polycystic ovary syndrome, and they cannot be used as a marker in clinical circumstances to evaluate the possible occurrence of polycystic ovary syndrome.

ABSTRACT The objective of this study was to elucidate the optimum protocol timing of thermal manipulation (TM) during embryogenesis, which underline genetic improvement of muscle thermotolerance acquisition. For the present study, 1,440 fertile eggs were divided randomly and equally into control (37.8 °C with 56% relative humidity) and four thermally manipulated groups (TM1, TM2, TM3, and TM4) subjected to 39 °C for 18 h with 65% relative humidity daily during different embryonic periods. Then, at day 35 post-hatch, all groups were subjected to thermal challenge at 43 °C for 6 h to identify the level of thermotolerance acquisition differences between them. Hsp70 mRNA expression was evaluated by using a relative quantitatively RT-qPCR. Single nucleotide polymorphisms sequence of the Hsp70 gene was evaluated by Sanger's sequencing method. Pectoral and thigh muscles samples were subjected to immunohistochemistry to detect Hsp70. Among TM conditions that were investigated, TM1 (39 °C for 18 h during embryonic days (ED) 7–11) induced a significant improvement in thermotolerance parameters (body temperature and T3 levels) during thermal challenge combined with an increase in the levels of Hsp70 mRNA and its protein with a high stability of nucleotide sequences in both pectoral and thigh muscles. The partial DNA sequence of Hsp70 gene in TM1 was reported, and nucleotide sequences were deposited in NCBI GenBank database with the accession numbers (MK852579) and (MK852580). Thigh muscle thermotolerance acquisition was higher than pectoral muscle during thermal challenge at 43 °C for 6 h. Thus, TM during ED7–11 may improve thermotolerance acquisition without adversely affecting performance.

ABSTRACT Incubating temperature and timing or duration is critical to determine the optimum protocol of thermal manipulation (TM), which underlines muscle growth improvement. Therefore, the aim of the present study is to determine the optimum period of embryonic TM that may result in the improvement of pectoral and thigh muscle myogenesis. This is done by investigating the level of mRNA expression of creatine kinase (CK) and lactate dehydrogenase (LDH). An additional goal is measuring the blood levels of CK and LDH as a biomarker of muscle injury due to the experimental thermal challenge on post-hatch day 35. The study was conducted on 1,440 fertile eggs (Ross broilers) that were divided randomly and equally into a control group and four treatment groups (TM1, TM2, TM3, and TM4). The treatment groups were daily subjected to TM at 39 ºC for 18h with 65% relative humidity (RH) during embryonic days (EDs) 7-11, 11-15, 15-18, and 7-18, respectively. Among the thermally manipulated groups that were investigated, TM1 (ED 7-11) resulted in significant improvement of mRNA expression and enzymatic concentration of CK and LDH in muscle during embryogenesis, as compared to the control. Six hours of TC showed the highest significant CK and LDH expression and concentration levels in the control as compared to TM groups. Thus, the results of this study indicate that TM during ED 7-11 improves pectoral and thigh muscles’ response to heat stress without adversely affecting their performance. This finding could be used by commercial breeders to enhance local broiler production.

ABSTRACT The aim of this study was to determine the optimum timing of thermal manipulation during early, mid, late and long lasting embryogenesis that may result in improvement of body performance and myofiber development (fiber diameter and thickness of perimysium) of pectoral and thigh muscles by a comparative morphometrical investigation on post-hatch day 35. 1440 fertile Ross broiler eggs were divided randomly and equally into 5 treatment groups: control (no TM), TM1, TM2 TM3 and TM4 were thermally subjected to 39ºC for 18h with 65% relative humidity daily during embryonic days ED 7-11, ED 11-15, ED 15-18 and ED 7-18 respectively. Out of TM conditions that were investigated, TM1 resulted in a significant improvement of hatchability rate (95.14%)and body performance when compared to the control. On post-hatch day 35, there were significant differences in BW, skinned carcass, breast, thigh and gut weight for all groups when compared to each other with a higher significant in TM1 and TM2 when compared to those of the control. FCR in this study was lower in TM groups when compared to the control group. In TM3 and TM4 the chicks show a significantly lower value of body performance when compared with those of the control group, TM1 and TM2.The histological evaluation of Pectoral muscle revealed myofibers of different diameter (38.39-40.27 µm) and perimysial thickening (17-30.5 µm), while myofibers diameter of thigh muscle ranged between (29-32.9 µm) and the perimysial thickening ranged between (14.36-22.15 µm). Thus, these results indicate that TM duringED7-11 improved muscle growth and body performance and this finding may be applied by commercial breeders to produce more enhanced broiler chickens.

ABSTRACT The objectives of current study were to evaluate the effect of botanical extracts as antioxidants on amino - and fatty acids profile of broiler meat. In total, 200 one-day-old broiler chicks were divided according to a completely randomized design into eight dietary treatments with five replicates of five chicks each. Dietary treatments were: A - positive control (PC) with antibiotics and synthetic antioxidants, B - negative control (NC), C - NC+Trigonellafoenum-graecum, D -NC+Nigella sativa seed, E -NC+Violaodorata, F -NC+Trachyspermumammi, G -PC+Trachyspermumammi+Trigonellafoenum-graecum+Violaodorata, and H -NC+ Trachyspermumammi, Trigonellafoenum-graecum and Viola odorata. Meat amino - and fatty acid contents were analyzed after slaughter at 35 days of age. Data were submitted to one-way analysis of variance and means were compared by Duncan’s Multiple Range test. The levels of histidine, isoleucine, leucine, lysine and threonine were significantly higher (p≤0.05) in the H, F, E, H and F groups respectively. Arginine, tryptophan, valine, methionine, and phenylalanine were significantly higher (p≤0.05) in the C, F, E, G and H groups. Total essential amino acid levels were highest (40.36%) in C group (p≤0.05). The meat of H broilers presented significantly higher (p≤0.05) unsaturated fatty acids (54.8%) and omega-3 contents (2.77%) and the lowest saturated fatty acid content (44.8%). Group F present the highest linoleic acid and omega-6 levels and group H the highest (p≤0.05) arachidonic and eicosapentaenoic acid (EPA) contents. It was concluded that botanical extracts aid health-beneficial meat production and the simultaneous inclusion of Trachyspermumammi, Trigonellafoenum-graecum, and Viola odorata in broiler diets is a more suitable option than the other dietary regimes.

A new analytical approach was developed involving cloud point extraction (CPE) and spectrofluorimetric determination of triamterene (TM) in biological fluids. A urine or plasma sample was prepared and adjusted to pH 7, then TM was quickly extracted using CPE, using 0.05% (w/v) of Triton X-114 as the extractant. The main factors that affected the extraction efficiency (the pH of the sample, the Triton X-114 concentration, the addition of salt, the extraction time and temperature, and the centrifugation time and speed) were studied and optimized. The method gave calibration curves for TM with good linearities and correlation coefficients (r) higher than 0.99. The method showed good precision and accuracy, with intra- and inter-assay precisions of less than 8.50% at all concentrations. Standard addition recovery tests were carried out, and the recoveries ranged from 94.7% to 114%. The limits of detection and quantification were 3.90 and 11.7 µg L-1, respectively, for urine and 5.80 and 18.0 µg L-1, respectively, for plasma. The newly developed, environmentally friendly method was successfully used to extract and determine TM in human urine samples.

BACKGROUND: Microorganisms growing in a biofilm are associated with chronic and recurrent human infections and are highly resistant to antimicrobial agents. There are various methods to detect biofilm production like Tissue Culture Plate (TCP), Tube method (TM), Congo Red Agar method (CRA), bioluminescent assay, piezoelectric sensors, and fluorescent microscopic examination. OBJECTIVE: This study was conducted to compare three methods for the detection of biofilms. METHOD: The study was carried out at the Department of Microbiology, Army Medical College, National University of Sciences and Technology, Pakistan, from January 2010 to June 2010. A total of 110 clinical isolates were subjected to biofilm detection methods. Isolates were identified by standard microbiological procedures. Biofilm detection was tested by TCP, TM and CRA. Antibiotic susceptibility test of biofilm producing bacteria was performed by using the Kirby-Bauer disc diffusion technique according to CLSI guidelines. RESULTS: The TCP method was considered to be superior to TM and CRA. From the total of 110 clinical isolates, TCP method detected 22.7% as high, 41% moderate and 36.3% as weak or non-biofilm producers. We have observed higher antibiotic resistance in biofilm producing bacteria than non-biofilm producers. CONCLUSION: We can conclude from our study that the TCP method is a more quantitative and reliable method for the detection of biofilm forming microorganisms as compared to TM and CRA methods, and it can be recommended as a general screening method for detection of biofilm producing bacteria in laboratories.

Estudos termodinâmicos relativos ao efeito da temperatura (de 27 a 47 ºC) sobre a estrutura da b-lactoglobulina-A bovina (BLG-A), em solução de cloreto de sódio 50 mmol L-1, na presença e ausência de íons Cr(III), usando espectroscopia de absorção no UV-Visível, dicroísmo circular (DC), próximo e distante e, espectroscopia de fluorescência, foram realizados. Os estudos com CD UV-distante, a diferentes temperaturas e na presença e ausência de íons Cr(III) não mostraram alteração significativa na estrutura secundária da proteína. Ao contrário, os estudos empregando DC UV-próximo evidenciaram alterações na estrutura terciária da BLG natural com o aumento da temperatura, indicada pela exposição de fragmentos de tirosina (Tyr). Estudos de espectroscopia de fluorescência em BLG-A natural, na ausência e presença de íons Cr(III) apresentam considerável alteração na estrutura terciária da proteína, devido ao aumento da temperatura. Com a estabilização da proteína na presença de íons Cr(III), a estrutura terciária da BLG revela considerável alterações a 37 e 47 °C as quais são concordantes com o aumento nos valores de Tm nestas temperaturas.

Thermodynamic studies of the effect of temperature (27-47 ºC) on the structure of bovine b-lactoglobulin-A (BLG-A) in the absence and presence of Cr(III) containing 50 mmol L-1 sodium chloride have been carried out using UV-Visible absorption spectroscopy, far and near circular dichroism (CD) and fluorescence spectroscopy. The far-UV CD studies do not show any significant change in the secondary structure of the protein at different temperatures in the absence and presence of Cr(III). On the contrary, the near-UV CD studies show change in the tertiary structure of the native BLG on increasing the temperature, indicating the exposure of Tyr residues. Fluorescence spectroscopic studies on the native BLG-A in the absence and presence of Cr(III) represent considerable change in the tertiary structure of the protein due to the increase in temperature. Due to protein stabilization in the presence of Cr(III) ions, the tertiary structure of BLG represents considerable alterations at 37 and 47 ο C that is in agreement with increasing of Tm values at these temperatures.

Um modelo isolado de coração de porco perfundido com sangue foi adaptado para o uso de fósforo 31 ressonância nuclear magnética (31 pRNM) em estudos de espectroscopia do metabolismo cardíaco durante cardioplegia sangüínea contínua e normotérmica (CSCN). O experimento foi dividido em dois grupos: Grupo I (n=5): os corações foram submetidos a 1 hora de CSCN. Grupo II (n=5): um período de 20 minutos de isquemia normotérmica durante o período de 1 hora de CSCN 17. A função do ventrículo esquerdo (VE) foi avaliada, com o coração batendo, utilizando um balão intraventricular, antes do período de parada cardioplégica e a seguir, quando o coração foi novamente perfundido com sangue normokalêmico. Durante todo o protocolo, análise espectroscópica do metabolismo cardíaco foi obtida utilizando-se 4.7 T/30cm Bruker TM Biospec 31 p RNM com uma resolução de 2 minutos para cada resultado. Ao final dos experimentos biópsias miocárdicas foram obtidas para análise de ATP e fosfocreatina (PCr) utilizando cromatografia liqüida de alta "performance" (HPLC). Não houve perda significante de ATP e PCr durante o período de parada cardioplégica com CSCN (Grupo I). Contudo, no Grupo II, a análise espectroscópica demonstrou perda completa de PCr após 14 ± 2 minutos durante a isquemia normotérmica acompanhada de aumento de fosfato inorgânico (Pi) e diminuição do pH intracelular. Quando reperfundido com CSCN, PCr, pH e Pi retornaram aos valores normais em 3 minutos. A função do VE avaliada através da elastância sistólica final foi mantida em 100 ± 10% dos valores obtidos antes da parada cardioplégica no Grupo I. No Grupo II, a função do VE foi de 88 ± 7% (p<0,05) dos valores precedentes à parada cardioplégica. Os resultados das biópsias do miocárdio demonstraram manutenção dos níveis normais de ATP (24 ± 3 µmol/g dry weight) e PCr (55 ± 14 µmol/g dry weight) no Grupo I, porém no Grupo II, apesar dos níveis normais de PCr após a reperfusão, os níveis de ATP diminuíram para 21 ± 3 µmol/g dry weight ao final do experimento. Este experimento demonstrou que o metabolismo cardíaco deteriorou após 14 minutos de isquemia normotérmica com conseqüente diminuição da função do VE (Grupo II). Porém, quando a cardioplegia sangüínea contínua e normotérmica foi utilizada sem interrupções (Grupo I) o metabolismo cardíaco e a função de VE foram preservadas, sugerindo que essa é a maneira ideal para se proteger o coração durante cirurgia cardíaca.

An isolated, blood perfused pig heart model was adapted for metabolic studies using 31 P NMR spectroscopy during continuous normothermic blood cardioplegia (CNBC). The experiment comprised two groups: In Group I (n=5) the hearts were subjected to 1 hour of CNBC. In Group II (n=5) a 20 min period of ischemia was introduced in the middle of the period of CNBC. Left ventricular function was assessed, in the beating heart, prior to the period of CNBC and after 60 min of cardioplegia, using an intra-ventricular balloon. During the entire period of the experiment, spectra were obtained using a 4.7 T/30 cm bore BrukerTM Biospec 31P NMR spectrometer with 2 min of resolution. At the end of the experiments myocardial biopsies were taken for ATP and phosphocreatine (PCr) analyses using high performance liquid chromatography (HPLC). ATP and PCr were maintained after the period of cardioplegic arrest (Group I). However the Group II, complete loss of PCr was shown after 14 ± 2 min of normothermic ischemia, followed by an increase of inorganic phophate (Pi) and decrease of intracellular pH. Following reperfusion after CNBC, PCr, pH and Pi returned normal values in 3 min. Left ventricular function, assessed by end-systolic elastance, was maintained at 100±10% of control in Group I. In the Group II, left ventricular function was 88 ± 7% (p<0.05) of the control. HPLC analyses of the myocardial biopsies showed normal values for Group I, ATP (24 ± 3 µmol/g dry weight) and PCr (55 ± 14 µmol/g dry weight), but in Group II, despite normal levels of PCr after reperfusion, the ATP levels decreased to 21 ± 3 µmol/g dry weight at the end of the experiments. These results showed that cardiac metabolism was seriously compromised after 14 min of normothermic ischemia and left ventricular function had decreased (Group II). However in Group I, in which CNBC was used without interruptions, function and metabolism were preserved, suggesting this is the ideal technique to protect the heart during cardiac surgery.