Engineering gain-of-function mutants of a WW domain by dynamics and structural analysis.

Proteins gain optimal fitness such as foldability and function through evolutionary selection. However, classical studies have found that evolutionarily designed protein sequences alone cannot guarantee foldability, or at least not without considering local contacts associated with the initial folding steps. We previously showed that foldability and function can be restored by removing frustration in the folding energy landscape of a model WW domain protein, CC16, which was designed based on Statistical Coupling Analysis (SCA). Substitutions ensuring the formation of five local contacts identified as "on-path" were selected using the closest homolog native folded sequence, N21. Surprisingly, the resulting sequence, CC16-N21, bound to Group I peptides, while N21 did not. Here, we identified single-point mutations that enable N21 to bind a Group I peptide ligand through structure and dynamic-based computational design. Comparison of the docked position of the CC16-N21/ligand complex with the N21 structure showed that residues at positions 9 and 19 are important for peptide binding, whereas the dynamic profiles identified position 10 as allosterically coupled to the binding site and exhibiting different dynamics between N21 and CC16-N21. We found that swapping these positions in N21 with matched residues from CC16-N21 recovers nature-like binding affinity to N21. This study validates the use of dynamic profiles as guiding principles for affecting the binding affinity of small proteins.

Effect of temperature manipulation during incubation on body weight, plasma parameters, muscle histology, and expression of myogenic genes in breast muscle of embryos and broiler chickens from two commercial strains.

1. This study evaluated the effect of a higher incubation temperature on body weight, plasma profile, histology and expression of myogenin (MYOG), insulin-like growth factor-I (IGF-I) and vascular endothelial growth factor A (VEGFA) genes in breast muscle of embryos and broilers from two commercial strains.2. A total of 784 eggs from Ross 308 and Cobb 500 broiler breeder flocks were used. Half of the eggs per strain were incubated at control temperature (37.8°C), whereas the other half were exposed to heat treatment (HT) of 38.8°C between embryonic day (ED) 10 and 14, for 6 h/day. Embryos and chicks were sampled on ED 19 and at hatch. A total of 480, one-day-old chicks per strain and incubation temperature were reared up to 42 d post-hatch.3. The HT increased hatch weight of Ross chicks and 42-d body weight of broilers from both strains. Lower plasma triacylglycerol levels were measured for HT embryos and broilers on ED 19 and 42 d post-hatch, respectively. HT reduced plasma T3 levels in Ross embryos and broilers for the same periods. Hepatic TBARS concentrations were elevated by HT compared to the control incubation.4. The HT reduced breast muscle VEGFA gene expression of Cobb embryos on ED 19, whereas expression was stimulated in day-old chicks. At 42 d post-hatch, fibre area was increased by HT regardless of strain. Compared to the control incubation, HT increased the breast yield of Ross broilers and leg yield of Cobb. Ross-HT broilers had a higher pH at 24 h after slaughter and better water holding capacity than Cobb-HT broilers.5. These results suggested that HT increased body weight, fibre area, IGF-I gene expression and lowered plasma triacylglycerol levels of broiler chickens from both strains at 42 d. However, HT influenced the expression of VEGF-A and MYOG genes and meat quality differently between the broiler strains.

Short communication: Identifying key parameters for modelling the impacts of livestock health conditions on greenhouse gas emissions.

Improved animal health can reduce greenhouse gas (GHG) emissions intensity in livestock systems while increasing productivity. Integrated modelling of disease impacts on farm-scale emissions is important in identifying effective health strategies to reduce emissions. However, it requires that modellers understand the pathways linking animal health to emissions and how these might be incorporated into models. A key barrier to meeting this need has been the lack of a framework to facilitate effective exchange of knowledge and data between animal health experts and emissions modellers. Here, these two communities engaged in workshops, online exchanges and a survey to i) identify a comprehensive list of disease-related model parameters and ii) test its application to evaluating models. Fifty-six parameters were identified and proved effective in assessing the potential of farm-scale models to characterise livestock disease impacts on GHG emissions. Easy wins for the emissions models surveyed include characterising disease impacts related to feeding.

Effects of 2100 MHz radio frequency radiation on ductus epididymis tissue in rats.

PURPOSE: The use of mobile phones is widespread since the beginning of 1990s. A great debate exists about the possible damage that the Radio Frequency - RF radiation from mobile phones exerts on different organs. The objective of this study was to investigate the possible histopathological effects of 2100 MHz RF radiation on rat ductus epididymis tissue using a light microscopy and immunohistochemical method after one or two month exposure. MATERIALS AND METHODS: The study was performed on 36 adult Wistar albino rats. 2100 MHz RF radiation was used with a specific absorption rate (SAR) of 0.36 W/kg for 30 min/day, 6 days per week for one or two months. There were 3 groups (n = 6 for each group): one month RF exposed group, two months RF exposed group, and the control group. RESULTS: At the end of the study, the structural changes in ductus epididymis tissue were evaluated. In both 2100 MHz RF exposed groups, the rat ductus epididymis sperm were not observed in some channels, a reduction in sperm density in some of the channels drew an attention. The loss of connective tissue and edematous areas were observed in cross channel interstitial connective tissue. In addition, it was observed that vascularization was highly increased with respect to the control group in cross-channel interstitial connective tissue. CONCLUSION: 2100 MHz RF exposure resulted in some structural changes in the male genital ducts of rats (Tab. 1, Fig. 5, Ref 20).

Adenine nucleotide translocase is acetylated in vivo in human muscle: Modeling predicts a decreased ADP affinity and altered control of oxidative phosphorylation.

Proteomics techniques have revealed that lysine acetylation is abundant in mitochondrial proteins. This study was undertaken (1) to determine the relationship between mitochondrial protein acetylation and insulin sensitivity in human skeletal muscle, identifying key acetylated proteins, and (2) to use molecular modeling techniques to understand the functional consequences of acetylation of adenine nucleotide translocase 1 (ANT1), which we found to be abundantly acetylated. Eight lean and eight obese nondiabetic subjects had euglycemic clamps and muscle biopsies for isolation of mitochondrial proteins and proteomics analysis. A number of acetylated mitochondrial proteins were identified in muscle biopsies. Overall, acetylation of mitochondrial proteins was correlated with insulin action (r = 0.60; P < 0.05). Of the acetylated proteins, ANT1, which catalyzes ADP-ATP exchange across the inner mitochondrial membrane, was acetylated at lysines 10, 23, and 92. The extent of acetylation of lysine 23 decreased following exercise, depending on insulin sensitivity. Molecular dynamics modeling and ensemble docking simulations predicted the ADP binding site of ANT1 to be a pocket of positively charged residues, including lysine 23. Calculated ADP-ANT1 binding affinities were physiologically relevant and predicted substantial reductions in affinity upon acetylation of lysine 23. Insertion of these derived binding affinities as parameters into a complete mathematical description of ANT1 kinetics predicted marked reductions in adenine nucleotide flux resulting from acetylation of lysine 23. Therefore, acetylation of ANT1 could have dramatic physiological effects on ADP-ATP exchange. Dysregulation of acetylation of mitochondrial proteins such as ANT1 therefore could be related to changes in mitochondrial function that are associated with insulin resistance.

BP-Dock: a flexible docking scheme for exploring protein-ligand interactions based on unbound structures.

Molecular docking serves as an important tool in modeling protein-ligand interactions. However, it is still challenging to incorporate overall receptor flexibility, especially backbone flexibility, in docking due to the large conformational space that needs to be sampled. To overcome this problem, we developed a novel flexible docking approach, BP-Dock (Backbone Perturbation-Dock) that can integrate both backbone and side chain conformational changes induced by ligand binding through a multi-scale approach. In the BP-Dock method, we mimic the nature of binding-induced events as a first-order approximation by perturbing the residues along the protein chain with a small Brownian kick one at a time. The response fluctuation profile of the chain upon these perturbations is computed using the perturbation response scanning method. These response fluctuation profiles are then used to generate binding-induced multiple receptor conformations for ensemble docking. To evaluate the performance of BP-Dock, we applied our approach on a large and diverse data set using unbound structures as receptors. We also compared the BP-Dock results with bound and unbound docking, where overall receptor flexibility was not taken into account. Our results highlight the importance of modeling backbone flexibility in docking for recapitulating the experimental binding affinities, especially when an unbound structure is used. With BP-Dock, we can generate a wide range of binding site conformations realized in nature even in the absence of a ligand that can help us to improve the accuracy of unbound docking. We expect that our fast and efficient flexible docking approach may further aid in our understanding of protein-ligand interactions as well as virtual screening of novel targets for rational drug design.

The antiviral lectin cyanovirin-N: probing multivalency and glycan recognition through experimental and computational approaches.

CVN (cyanovirin-N), a small lectin isolated from cyanobacteria, exemplifies a novel class of anti-HIV agents that act by binding to the highly glycosylated envelope protein gp120 (glycoprotein 120), resulting in inhibition of the crucial viral entry step. In the present review, we summarize recent work in our laboratory and others towards determining the crucial role of multivalency in the antiviral activity, and we discuss features that contribute to the high specificity and affinity for the glycan ligand observed in CVN. An integrated approach that encompasses structural determination, mutagenesis analysis and computational work holds particular promise to clarify aspects of the interactions between CVN and glycans.

Electrostatic effect of the ribosomal surface on nascent polypeptide dynamics.

The crucial molecular events accompanying protein folding in the cell are still largely unexplored. As nascent polypeptides emerge from the ribosomal exit tunnel, they come in close proximity with the highly negatively charged ribosomal surface. How is the nascent polypeptide influenced by the ribosomal surface? We address this question via the intrinsically disordered protein PIR and a number of its variably charged mutants. Two different populations are identified: one is highly spatially biased, and the other is highly dynamic. The more negatively charged nascent polypeptides emerging from the ribosome are richer in the extremely dynamic population. Hence, nascent proteins with a net negative charge are less likely to interact with the ribosome. Surprisingly, the amplitude of the local motions of the highly dynamic population is much wider than that of disordered polypeptides under physiological conditions, implying that proximity to the ribosomal surface enhances the molecular flexibility of a subpopulation of the nascent protein, much like a denaturing agent would. This effect could be important for a proper structural channeling of the nascent protein and the prevention of cotranslational kinetic trapping. Interestingly, a significant population of the highly spatially biased nascent chain, probably interacting extensively with the ribosome, is present even for very negatively charged nascent proteins. This "sticking" effect likely serves to protect nascent proteins (e.g., from cotranslational aggregation). In all, our results highlight the influence of the ribosome in nascent protein dynamics and show that the ribosome's function in protein biogenesis extends well beyond catalysis of peptide bond formation.

Adaptive support ventilation for faster weaning in COPD: a randomised controlled trial.

Adaptive support ventilation (ASV) is a closed-loop ventilation mode that can act both as pressure support ventilation (PSV) and pressure-controlled ventilation. Weaning with ASV shows promising results, mainly in post-cardiac surgery patients. The aim of the present randomised controlled study was to test the hypothesis that weaning with ASV could reduce the weaning duration in patients with chronic obstructive pulmonary disease (COPD) when compared with PSV. From among 435 COPD patients admitted to the intensive care unit (ICU) during a 20-month period, 97 were enrolled. Patients were assigned at random to either ASV or PSV as a weaning mode. Compared with PSV, ASV provided shorter weaning times (median 24 (interquartile range 20-62) h versus 72 (24-144) h, p=0.041) with similar weaning success rates (35 out of 49 for ASV and 33 out of 48 for PSV). Length of stay in the ICU was also shorter with ASV but the difference was not statistically significant. This study suggests that ASV may be used in the weaning of COPD patients with the advantage of shorter weaning times. Further studies are needed to investigate the role and potential advantages of ASV in the weaning period of different patient groups.

Comparison of topical 3% diclofenac sodium gel and 5% imiquimod cream for the treatment of actinic keratoses.

BACKGROUND: There is a wide spectrum of treatments available for actinic keratosis (AK). Topical diclofenac sodium and imiquimod are two topical treatments, which are noninvasive, easily applied, well-tolerated and effective. AIM: To compare the effects of topical 3% diclofenac sodium plus hyaluranon (DFS) gel, 5% imiquimod (IMQ) cream, and base cream (BC) in patients with AK. METHODS: In total, 61 patients, diagnosed clinically and histopathologically as having AK, were randomized into three treatment groups to receive topical treatment with either DFS (twice daily for 12 weeks), IMQ (twice per week for 16 weeks) or BC (twice daily for 12 weeks). Patients were evaluated clinically at 0, 4, 8, 12, 16, 20 and 24 weeks. Treatment efficacy was assessed by Total Thickness Score (TTS) and Patient Global Improvement Index (PGII). RESULTS: Complete clearance rates for DFS, IMQ and BC at the end of the treatment and at the end of the total follow-up period were 19.1%, 20% and 0%, and 14.3%, 45% and 0%, respectively. Although the average TTS value of the DFS group at week 24 was significantly higher than that of the IMQ group, the PGII values were not significantly different. CONCLUSIONS: Although DFS and IMQ each had considerable efficacy in the treatment of AK, the efficacy of DFS seemed to decrease after cessation of treatment.

Comparison of intrathecal hyperbaric and isobaric levobupivacaine in urological surgery.

AIM: The aim of our study was to compare the efficacy of hyperbaric and isobaric solutions of intrathecal levobupivacaine for transurethral endoscopic surgery. METHODS: Urological patients who were scheduled for elective surgery under spinal anesthesia were enrolled. The heavy group received 13.5 mg of hyperbaric levobupivacaine, while the plain group received 13.5 mg isobaric levobupivacaine, both intrathecally in a 3 mL total volume. Sensory and motor block, hemodynamic parameters, pain scores, adverse effects, and analgesic requirements of the patients were recorded. RESULTS: Values of the time to onset of T10 sensory block, time to maximum sensory block, regression to L1 dermatome, time to motor block Bromage 1, time to motor block Bromage 3, and time to the end of motor block (Bromage 0) were all smaller in group 1 than in group 2 (all P values <0.05). No difference between the groups with regard to time to two segment regression of sensory block could be detected. The mean duration of initial analgesic effect, extent of maximal block, and side effects were the same in both groups (P>0.05). CONCLUSIONS: We concluded that the clinical efficacy of hyperbaric levobupivacaine was superior to the isobaric form in spinal anesthesia for transurethral resection.

Effect of melatonin and time of administration on irradiation-induced damage to rat testes.

The effect of ionizing irradiation on testes and the protective effects of melatonin were investigated by immunohistochemical and electron microscopic methods. Eighty-two adult male Wistar rats were divided into 10 groups. The rats in the irradiated groups were exposed to a sublethal irradiation dose of 8 Gy, either to the total body or abdominopelvic region using a 60Co source at a focus of 80 cm away from the skin in the morning or evening together with vehicle (20% ethanol) or melatonin administered 24 h before (10 mg/kg), immediately before (20 mg/kg) and 24 h after irradiation (10 mg/kg), all ip. Caspace-3 immunoreactivity was increased in the irradiated group compared to control (P < 0.05). Melatonin-treated groups showed less apoptosis as indicated by a considerable decrease in caspace-3 immunoreactivity (P < 0.05). Electron microscopic examination showed that all spermatogenic cells, especially primary spermatocytes, displayed prominent degeneration in the groups submitted to total body and abdominopelvic irradiation. However, melatonin administration considerably inhibited these degenerative changes, especially in rats who received abdominopelvic irradiation. Total body and abdominopelvic irradiation induced identical apoptosis and testicular damage. Chronobiological assessment revealed that biologic rhythm does not alter the inductive effect of irradiation. These data indicate that melatonin protects against total body and abdominopelvic irradiation. Melatonin was more effective in the evening abdominopelvic irradiation and melatonin-treated group than in the total body irradiation and melatonin-treated group.

Effect of melatonin and time of administration on irradiation-induced damage to rat testes

The effect of ionizing irradiation on testes and the protective effects of melatonin were investigated by immunohistochemical and electron microscopic methods. Eighty-two adult male Wistar rats were divided into 10 groups. The rats in the irradiated groups were exposed to a sublethal irradiation dose of 8 Gy, either to the total body or abdominopelvic region using a 60Co source at a focus of 80 cm away from the skin in the morning or evening together with vehicle (20% ethanol) or melatonin administered 24 h before (10 mg/kg), immediately before (20 mg/kg) and 24 h after irradiation (10 mg/kg), all ip. Caspace-3 immunoreactivity was increased in the irradiated group compared to control (P < 0.05). Melatonin-treated groups showed less apoptosis as indicated by a considerable decrease in caspace-3 immunoreactivity (P < 0.05). Electron microscopic examination showed that all spermatogenic cells, especially primary spermatocytes, displayed prominent degeneration in the groups submitted to total body and abdominopelvic irradiation. However, melatonin administration considerably inhibited these degenerative changes, especially in rats who received abdominopelvic irradiation. Total body and abdominopelvic irradiation induced identical apoptosis and testicular damage. Chronobiological assessment revealed that biologic rhythm does not alter the inductive effect of irradiation. These data indicate that melatonin protects against total body and abdominopelvic irradiation. Melatonin was more effective in the evening abdominopelvic irradiation and melatonin-treated group than in the total body irradiation and melatonin-treated group.

Placental expression of insulin-like growth factor-I, fibroblast growth factor-basic and neural cell adhesion molecule in pregnancies with small for gestational age fetuses.

OBJECTIVE: To investigate placental expression of insulin-like growth factor-I (IGF-I), fibroblast growth factor-basic (FGF-b) and neural cell adhesion molecule (N-CAM) regarding the pathogenesis of pregnancies with small for gestational age (SGA) fetuses. STUDY DESIGN: An immunohistochemical analysis using anti-IGF-I, anti-FGF-b and anti-N-CAM antibodies was carried out on 4% paraformaldehyde-fixed placental tissues of third trimester pregnancies complicated with SGA fetuses (n=12) and subjects exhibiting appropriately grown fetuses (n=10). Immunostaining patterns of chorionic villi and amniochorionic membranes were assessed. RESULT: IGF-I, FGF-b and N-CAM immunostainings in chorionic villi demonstrated significantly increased immunoreactivities in cytotrophoblasts of SGA cases, whereas increased IGF-I immunostaining in syncitiotrophoblasts and increased N-CAM immunostaining in capillary endothelium were noted in the same group. IGF-I, FGF-b and N-CAM immunostainings in amniochorionic membranes revealed significantly decreased IGF-I immunoreactivities in extravillous trophoblasts and increased IGF-I immunoreactivities in decidual cells of SGA cases, while significantly decreased N-CAM immunoreactivities in both decidual cells and extravillous trophoblasts were noted. FGF-b immunostaining revealed no significant differences in both extravillous trophoblasts and decidual cells of SGA cases. CONCLUSION: Increased placental expression of IGF-I, FGF-b and N-CAM may act in an autocrine and/or paracrine manner to restore the impaired trophoblastic proliferation, migration and metabolism at all gestational stages by means of a positive feedback mechanism.

The midterm results of cardiac transplantation patients.

OBJECTIVE: Cardiac transplantation is an important treatment option that increases the survival and decreases the limitations in effort capacity among patients with end-stage heart disease. In this study we have presented the midterm results of 13 patients who underwent cardiac transplantation between 2003 and 2007. PATIENTS AND METHODS: There were 10 male and three female patients of mean age of 32 +/- 13.27 years (12 to 54). In one patient, we performed combined cardiac and renal transplantation. Ischemic cardiac disease was present in six patients and cardiomyopathy in seven patients. The mean age of the donors was 23.3 +/- 11.8 years (12 to 46). Corticosteroids, cyclosporine, and mycophenolate mofetil were used for immunosuppression. Sirolimus was employed in five cases due to impaired renal function. Patients were followed by echocardiography, endomyocardial biopsy, and dobutamine stress echocardiography. RESULTS: The mean follow-up was 18.6 +/- 13.4 (1 to 38) months. In four patients, there was grade IIIA (II-R) rejection. In five patients, tacrolimus or cyclosporine was replaced with sirolimus due to elevated creatinine levels. Dobutamine stress echocardiography was positive in one patient, who displayed a severe left main coronary artery lesion. There was no operative mortality. There was only one hospital mortality (7.6%). Two patients died in the midterm. The overall mortality on follow-up was 3 (23.1%). The survival rates in the first, second, and third years were 92%, 88%, and 75%, respectively. Ejection fraction were more than 50%; all of posttransplant survivors showed good effort capacity. CONCLUSION: Cardiac transplantation is a definitive, safe, and effective treatment for patients with end-stage heart failure.

Successful cardiac transplantation from donor with carbon monoxide intoxication: a case report.

BACKGROUND: The use of hearts for cardiac transplantation from donors with brain death due to exposure to high concentrations of carbon monoxide is still under discussion. In this short report we have presented a patient who underwent a successful cardiac transplantation from a brain-dead donor who had cardiopulmonary resuscitation after carbon monoxide intoxication. METHOD: A standard biatrial anastomosis technique was used in our patient. The transplantation was uneventful with donor ischemic time of 180 minutes. The patient was treated with mechanical ventilation for 72 hours. The donor liver biopsy during harvesting did not reveal irreversible changes. Although the donor had a history of cardiopulmonary resuscitation, the left ventricular ejection fraction was 55% and the echocardiographic evaluation revealed normal cardiac contractions with acceptable hemodynamic parameters. Positive inotropic support was needed in the early postoperative period. We did not observe any changes related to intoxication in the endomyocardial biopsy. CONCLUSIONS: We concluded that successful heart transplantation can be performed using hearts from patients succumbing to carbon monoxide poisoning in the presence of adequate cardiac functional parameters. This group will increase the number of cardiac transplantations and decrease the incidence of deaths among patients on transplantation lists.

Morbilidad psiquiátrica y calidad de vida en pacientes con urticaria crónica idiopática / Psychiatric morbidity and quality of life in patients with chronic idiopathic urticaria

Objetivos. Determinar la prevalencia de la morbilidad psiquiátrica en los pacientes con urticaria crónica idiopática (UCI); determinar la calidad de vida relacionada con la salud (HRQL) en los pacientes con UCI en comparación con controles sanos. Métodos. Se seleccionaron 350 pacientes con urticaria crónica, entre el 1/01/2005 y 31/04/2005 en el Departamento de Alergia de la Universidad de Estambul, que cumplieran con los criterios de inclusión: pacientes con diagnóstico de UCI, con edad de entre 18 y 65 años, sin síntomas psicóticos ni retardo mental, con capacidad cognitiva normal y ausencia de enfermedad comórbida. Como control se eligió un grupo de individuos sanos con características demográficas comunes. Todos los participantes recibieron información sobre el estudio mediante consentimiento informado. Se realizó el diagnóstico de UCI descartando, para ello, todas las causas posibles (drogas, químicos, alimentos) mediante dietas de eliminación y con prick tests negativos. Además, se realizaron exámenes de laboratorio: hematología, enzimas hepáticas, hormonas tiroideas, anticuerpos antinucleares y test del suero autólogo. Se utilizó una escala visual analógica, además de entrevistas clínicas para medir la severidad de la enfermedad. Por otra parte, la evaluación psiquiátrica consistió en: entrevistas de forma semiestructurada en donde los participantes completaron un cuestionario de HRQL (resultado de la encuesta de salud) de 36 ítems de forma corta (SF-36) y además una entrevista clínica estructurada por el eje DSM-IV para diagnóstico de enfermedad psiquiátrica en los pacientes con UCI. Resultados. En este estudio se incluyó a 84 pacientes con UCI y 75 controles sanos. La media de edad ± desvío estándar fue 36,83 ± 10,26. El 84% de los pacientes eran mujeres; la duración media (±DE) de la enfermedad fue 6,34 ± 7,2 años y los síntomas fueron intermitentes en el 51% de los pacientes.

Dietary vitamin E (alpha-tocopherol acetate) and selenium supplementation from different sources: performance, ascites-related variables and antioxidant status in broilers reared at low and optimum temperatures.

1. This study compared the effect of dietary supplementation with organic or inorganic selenium (Se) sources plus control amounts or large amounts of vitamin E (alpha-tocopherol acetate) in broilers raised at control (20 to 24 degrees C) or low (14.5 to 16.8 degrees C) temperatures after 2 weeks of age. 2. The following dietary treatments were used from one day old. Diet 1, the control diet, comprised a commercial diet containing 0.15 mg/kg inorganic Se and 50 mg vitamin E/kg feed. Diet 2 was the same as diet 1, supplemented with 0.15 mg/kg inorganic Se. Diet 3 was the same as diet 2 but was supplemented with 200 mg/kg vitamin E. Diet 4 was the same as diet 1, but inorganic Se was replaced with 0.30 mg/kg organic Se. Diet 5 was the same as diet 4, supplemented with 200 mg/kg vitamin E. 3. Low temperature reduced the growth rate of broilers; however, at 6 weeks, there were no differences in the body weights of birds fed on organic Se supplemented diets housed at low or control temperature. The feed conversion ratio was significantly affected by low temperature but not by diet. The heterophil/lymphocyte ratio was higher in chicks after one week in the cold, indicating mild stress. Blood triiodothyronine levels were significantly higher in birds after 1 and 4 weeks in the cold but thyroxin was not affected. 4. Organic Se supplementation increased relative lung weight at the control temperature, which might lead to greater respiratory capacity. Relative spleen weight significantly decreased in broilers fed diets supplemented with inorganic Se under cold conditions, a possible indication of chronic oxidative stress. 5. At the low temperature, supplementation with organic Se alone, or with inorganic Se and vitamin E increased glutathione peroxidase (GSHPx) activity and glutathione (GSH) concentration in the liver of broilers, which may indicate increased activity of birds' antioxidant defence against suboptimal environments.

Locally advanced breast carcinoma treated with neoadjuvant chemotherapy: are the changes in serum levels of YKL-40, MMP-2 and MMP-9 correlated with tumor response?

Serum levels of YKL-40, MMP-2 and MMP-9 in 27 patients with locally advanced breast carcinoma received neoadjuvant chemotherapy were measured. All patients underwent neoadjuvant chemotherapy named as FAC protocol (5-Fluorouracil, Doxorubicin and Cyclophosphamide) with 21 days interval. There was 26,7% decrease in mean serum YKL-40 levels (from 146,4 microg/ml to 107,3 microg/ml) in clinically responsive group. This level was almost unchanged in non-responsive group (P>0, 05). There was 42, 1% decrease in mean serum YKL-40 levels (from 173,1 microg/ml to 98, 8 microg/ml) in pathologically responsive group. This decrease was more dramatic in patients with complete pathological response (70, 2%). However, this level was slightly increased in non-responsive group. Changes in serum levels of MMP-2 and MMP-9 were not found to be associated with tumor response. Serum measurement of YKL-40 can be a helpful tool to predict pathological tumor response in breast cancer patients with neoadjuvant chemotherapy but not MMP-2 and MMP-9.