Search

Tiliroside is a glycosidic flavonoid present in many plants species including Helicteres velutina K. Schum (Malvaceae sensu lato), commonly known in Brazil as “pitó”. This molecule has been shown to have many biological activities, however no study has been carried out to investigate the toxicity of this substance. The present work aimed to evaluate the possible cellular toxicity in silico, in vitro and ex-vivo of the kaempferol-3-O-β-D-(6”-E-p-coumaroyl) glucopyranoside (tiliroside), through chemical structure analysis, toxicity assessment and predictive bioactive properties, using human samples for in vitro and ex-vivo tests. The in silico analysis suggests that tiliroside exhibited great absorption index when penetrating biological membranes. In addition, it also displayed considerable potential for cellular protection against free radicals, and anticarcinogenic, antioxidant, antineoplastic, anti-inflammatory, anti-hemorrhagic and antithrombotic activities. The assessment of the hemolytic and genotoxic effects of tiliroside showed low hemolysis rates in red blood cells and absence of cellular toxicity in the oral mucosa cells. The data obtained indicate that this molecule could be a promising therapeutic approach as a possible new drug with biotechnological potential. Resumo O tilirosídeo é um flavonóide glicosídico presente em muitas espécies de plantas, incluindo Helicteres velutina K. Schum (Malvaceae sensu lato), conhecida no Brasil como “pitó”. Esta molécula mostrou ter muitas atividades biológicas, porém nenhum estudo foi realizado para investigar a toxicidade dessa substância. O presente trabalho teve como objetivo avaliar a possível toxicidade celular in silico, in vitro e ex-vivo do kaempferol-3-O-β-D- (6 ”-Ep-coumaroil) glucopiranosídeo (tilirosídeo), por meio de análises de estrutura química, toxicidade avaliação e propriedades bioativas preditivas, utilizando amostras humanas para testes in vitro e ex-vivo. A análise in silico sugere que o tilirosídeo exibe bom índice de absorção para penetrar nas membranas biológicas. Além disso, apresentou considerável potencial de proteção celular contra os radicais livres e com atividades anticarcinogênica, antioxidante, antineoplásica, antiinflamatória, anti-hemorrágica e antitrombótica. A avaliação dos efeitos hemolíticos e genotóxicos do tilirosídeo mostrou baixas taxas de hemólise nas hemácias e ausência de toxicidade em células da mucosa oral. Os dados obtidos indicam que esta molécula pode possuir uma abordagem terapêutica promissora como uma possível nova droga com potencial biotecnológico.

Background Micronutrient deficiencies are identified among obese individuals. Vitamin D deficiency (VDD) is prevalent in obese children, and is hypothesized to cause insulin resistance and metabolic abnormalities. This study aimed to determine the effect of vitamin D supplementation on obesity and related metabolic abnormalities among obese Sri Lankan children with VDD. Methods A triple-blind randomized controlled trial was conducted among vitamin D deficient (n = 96), randomly allocated to three intervention arms - treatment arm receiving weekly vitamin D2 50,000 IU; supplementation arm receiving 2500 IU weekly and control arm, receiving placebo. Anthropometry, percentage fat mass (%FM) and blood pressure were assessed and fasting blood glucose, fasting insulin, lipid profile, aspartate transaminase (ALT), alanine transaminase (AST), vitamin D, parathyroid hormone (PTH) and hs-CRP and OGTT with 2-h random blood glucose and insulin was performed at baseline and after 24 weeks of treatment. Ethics Review Committee of Faculty of Medicine, University of Colombo approved the protocol. Results Waist circumference Z-score, %FM and serum calcium significantly improved across all three arms, ALT significantly improved in treatment and supplementation arms while, BMI Z-score, PTH and vitamin D significantly improved in the treatment arm. Biceps (p = 0.035) and subscapular (0.048) skin fold thickness, vitamin D (p = 0.004) and ALT (p = 0.012) significantly improved in the treatment arm. Conclusions A strict dietary and physical activity regimen could improve some of the anthropometric, body composition and metabolic profiles, but high dose vitamin D, enhances those improvements. Therefore high dose vitamin D seems to potentiate management outcomes of obese children with vitamin D deficiency. Trial registration The study was registered at the Sri Lanka Clinical Trials Registry (SLCTR/2015/017) on 12th September 2015 at https://slctr.lk/trials/slctr-2015-017.

Objective The aim of this study was to analyze whether ω-3 supplementation improves cardiometabolic profile in individuals with cardiovascular risk factors and to determine the effect of adiponectin levels on these changes. Methods In this double-blind, placebo-controlled, 2-mo clinical trial, we randomized 80 individuals of both sexes (mean age 52 y) with at least one cardiovascular risk factor (excess weight, hypertension, dyslipidemia, diabetes, or smoking) into two groups: ω-3 (supplemented with 3 g/d of fish oil containing 37% eicosapentaenoic acid and 23% docosahexaenoic acid) and placebo (3 g/d of sunflower oil containing 65% linoleic acid). At baseline and after the intervention, we evaluated serum adiponectin, leptin, lipid profile, apolipoproteins (apo), electronegative low-density lipoprotein (LDL[−]), and glucose metabolism (glucose and insulin). Results After supplementation, the ω-3 group showed an increase in serum adiponectin. After stratifying the ω-3 group by adiponectin concentration at baseline, participants with lower adiponectin concentration showed a higher reduction of total cholesterol, LDL, LDL/high-density lipoprotein ratio, LDL/apo B, and LDL(−). Individuals with a higher variation of adiponectin concentration after ω-3 supplementation presented with reduced blood glucose. The variation of serum adiponectin induced by ω-3 supplementation was negatively correlated with the Framingham and Adult Treatment Panel IV scores (r = −0.4 and P Conclusions Adiponectin is shown as one of the mechanisms by which ω-3 improves cardiometabolic profile in persons with cardiovascular risk. Moreover, the benefit varies according to the adiponectin basal level and adiponectin variation after supplementation.

The relationship between epilepsy and the presence of visceral larva migrans caused by Toxocara canis in Mexican children remains uncertain; however, this relationship needs to be elucidated because these parasite larvae can invade the human central nervous system. Accordingly, this study aimed to determine the frequency and specificity of anti-T. canis antibodies in the sera of children with epilepsy to determine the relationship between this parasite and epilepsy. The sera samples of 214 children were examined: 111 children diagnosed with epilepsy and 103 clinically healthy children without neurological disorders. In the sera of each group, the presence and specificity of anti-T. canis and anti-Ascaris lumbricoides antibodies, as well as the cross-reactivity between them, were assessed using enzyme-linked immunosorbent assay and Western blotting analysis. Among the children with epilepsy, 25.2% exhibited seropositivity to T. canis. Cross-reactivity against the A. lumbricoides antigen was present in 46.8% of the children with epilepsy, whereas 11.7% of the children with epilepsy and anti-T. canis antibodies did not exhibit cross-reactivity against this antigen. The Western blotting analysis of the sera from the children with epilepsy demonstrated the presence of T. canis proteins, with molecular weights of 24, 35, 55, 70, 120 and 210 kDa, and A lumbricoides proteins with molecular weights of 70, 80 and 110 kDa. Our results revealed the presence of anti-T. canis antibodies in the children with epilepsy; furthermore, cross-reactivity tests with A. lumbricoides showed the importance of the presence of anti-T. canis antibodies in revealing the relationship between this parasite and epilepsy in children.

The primary objective of this experiment was to test the hypothesis that concentrations of soluble (SDF), insoluble (IDF), and total dietary fiber (TDF) in feed ingredients used in diets for pigs and poultry analyzed using Method AOAC 2011.25 are greater than values determined using Method AOAC 991.43. A second objective was to determine the variation that may exist among 3 laboratories using the 2 methods with the AnkomTDF Dietary Fiber Analyzer (Ankom Technology, Macedon, NY). The 3 laboratories were the Ministry of Agriculture Feed Industry Center (MAFIC) at China Agricultural University, Trouw Nutrition, and Hans H. Stein Monogastric Nutrition Laboratory at University of Illinois at Urbana-Champaign (UIUC). All laboratories analyzed SDF and IDF in feed ingredients in duplicate or triplicate using both methods AOAC 991.43 and 2011.25 with the AnkomTDF Dietary Fiber Analyzer. The 9 test ingredients were wheat, soybean meal, rapeseed meal, sugar beet pulp, peas, horse beans, native pea starch, and 2 samples of corn; 1 from Europe and 1 from China. All ingredient samples, with the exception of Chinese corn, were procured by Trouw Nutrition, ground to pass through a 0.5 mm screen, subsampled, and sent to MAFIC and UIUC. Data were analyzed using SDF, IDF, and TDF as response variables, replication as random effect, and method and location as fixed effects over all ingredients and within each ingredient. When averaged among 9 different ingredients, results indicated that SDF, IDF, and TDF values were not different with either method or at any laboratory. However, the concentration of IDF in corn, wheat, peas, and sugar beet pulp determined using Method AOAC 991.43 was greater (P < 0.05) compared with 2011.25. Soluble dietary fiber determined using Method AOAC 2011.25 was greater (P < 0.05) in corn, rapeseed meal, soybean meal, and sugar beet pulp compared with 991.43. There was no difference in TDF determined with either method, except for wheat having greater (P < 0.05) TDF when determined using Method AOAC 991.43. Interlaboratory variation for SDF, IDF, and TDF was 0.38, 0.87, 1.20, respectively, with Method AOAC 991.43 and 0.40, 0.93, and 1.27, respectively, with 2011.25. Therefore, values determined with the AnkomTDF Analyzer are repeatable among laboratories and can be used in feed formulation worldwide. In conclusion, it is recommended that Method AOAC 991.43 be used to determine SDF, IDF, and TDF in feed ingredients and diets for pigs and poultry.

The African buffalo (Syncerus caffer) is considered the most important maintenance host of bovine tuberculosis (BTB) in wildlife in Southern Africa. The diagnosis of Mycobacterium bovis infection in this species mostly relies on the single intradermal comparative tuberculin test (SICTT). As an alternative, the BOVIGAM® 1G, an interferon-gamma (IFN-γ) release assay, is frequently used. The test performance of cell-mediated immunity (CMI-) and humoral immunity (HI-) based assays for the detection of M. bovis infections in buffaloes was compared to identify the test or test combination that provided the highest sensitivity in the study. Buffaloes were sampled during the annual BTB SICTT testing in the Hluhluwe-iMfolozi-Park (KwaZulu-Natal, South Africa) during June 2013. A total of 35 animals were subjected to the SICTT, 13 of these tested positive and one showed an inconclusive reaction. CMI-based assays (BOVIGAM® 1G (B1G) and BOVIGAM® 2G (B2G)) as well as a serological assay (IDEXX TB ELISA) were used to further investigate and compare immune responsiveness. Thirteen SICTT positive buffaloes and one inconclusive reactor were slaughtered and a post-mortem (PM) examination was conducted to confirm BTB. Lesions characteristic of BTB were found in 8/14 animals (57.1%). Test results of individual assays were compared with serial and parallel test interpretation and the sensitivity was calculated as a percentage of test positives out of the number of SICTT positive animals with granulomatous lesions (relative sensitivity). The B1G assay showed the highest individual sensitivity (100%; 8/8) followed by the B2G assay (75%; 6/8) and the IDEXX TB ELISA (37.5%; 3/8). Therefore, using in parallel interpretation, any combination with the B1G showed a sensitivity of 100% (8/8), whereas combinations with the B2G showed a 75% sensitivity (6/8). Out of the 21 SICTT negative animals, 7 animals showed responsiveness in the B2G or IDEXX TB ELISA. In conclusion, this study has shown that the BOVIGAM® IFN-γ assay had the highest test performance.

Recently, the trend of enhancing daily consumed foods with functional ingredients has been increasing. Bread, being a food product, which is consumed regularly, contains most nutrients but lacks antioxidant rich polyphenolic compounds due to the usage of refined wheat flour. Green coffee beans (GCB) are known for their green colour, which is the colour of coffee beans prior to roasting. The aroma is described as mild, green, and bean-like, which deters many from trying to consume the product. This experiment is conducted to determine antimicrobial properties and the sensory response to study the physiological effect of caffeine in bread incorporated with GCB. The bread was baked with three (3) variable parameters which were: the concentration of GCB powder, baking time, and baking temperature. The effect of concentration of GCB powder, baking time, and baking temperature on bacteria growth were also observed. It is concluded that to control the bacteria growth, the optimum condition of incorporated bread was determined at baking temperature of 180.3°C, baking time of 39.9 minutes, and GCB powder concentration of 4.85 wt%. Sensory evaluation analysis for the optimised bread (enriched with GCB) and control bread (without GCB) were conducted by a series of sensory judges. The sensory evaluation determined the differences in all aspects of appearance, aroma, taste, and texture between the enriched and normal bread. The most significant differences are that the bread incorporated with green coffee bean was darker in colour, has more distinctive wood, caramel, and smoke smell, more pungent and less salty. In addition, it is also more bitter, and harder with a stronger aftertaste when compared to normal bread. In conclusion, bread enriched with GCB exhibits strong antimicrobial properties and acceptable differences in appearance, aroma, taste, and texture compared to normal bread.

A number of feed additives are marketed to assist in boosting the pigs' immune system, regulate gut microbiota, and reduce negative impacts of weaning and other environmental challenges. The most commonly used feed additives include acidifiers, zinc and copper, prebiotics, direct-fed microbials, yeast products, nucleotides, and plant extracts. Inclusion of pharmacological levels of zinc and copper, certain acidifiers, and several plant extracts have been reported to result in improved pig performance or improved immune function of pigs. It is also possible that use of prebiotics, direct-fed microbials, yeast, and nucleotides may have positive impacts on pig performance, but results have been less consistent and there is a need for more research in this area. Keywords: Acidifiers, Direct-fed microbials, Minerals, Plant extracts, Prebiotics, Pigs

Conventional control and eradication strategies for bovine tuberculosis (BTB) face tremendous difficulties in developing countries; countries with wildlife reservoirs, a complex wildlife-livestock-human interface or a lack of veterinary and veterinary public health surveillance. Vaccination of cattle and other species might in some cases provide the only suitable control strategy for BTB, while in others it may supplement existing test-and-slaughter schemes. However, the use of live BCG has several limitations and the global rise of HIV/AIDS infections has furthermore warranted the exploration of inactivated vaccine preparations. The aim of this study was to compare the immune response profiles in response to parenteral vaccination with live BCG and two inactivated vaccine candidates in cattle. Twenty-four mixed breed calves (Bos taurus) aged 4–6 months, were allocated to one of four groups and vaccinated sub-cutaneously with live M. bovis BCG (Danish 1331), formalin-inactivated M. bovis BCG, heat-killed M. bovis or PBS/Montanide™ (control). Interferon-γ responsiveness and antibody production were measured prior to vaccination and at weekly intervals thereafter for twelve weeks. At nine weeks post-priming, animals were skin tested using tuberculins and MTBC specific protein cocktails and subsequently challenged through intranodular injection of live M. bovis BCG. The animals in the heat-killed M. bovis group demonstrated strong and sustained cell-mediated and humoral immune responses, significantly higher than the control group in response to vaccination, which may indicate a protective immune profile. Animals in this group showed reactivity to the skin test reagents, confirming good vaccine take. Lastly, although not statistically significant, recovery of BCG after challenge was lowest in the heat-killed M. bovis group. In conclusion, the parenteral heat-killed M. bovis vaccine proved to be clearly immunogenic in cattle in the present study, urging further evaluation of the vaccine in challenge studies using virulent M. bovis and assessment of vaccine efficacy in field conditions., This work was supported by NWO-WOTRO Science for Global Development (grant W01.65.321.00) to VR & AL (https://www.nwo.nl/en/about-nwo/organisation/nwo-domains/wotro), and NWO-WOTRO funding was used towards all experimental costs, including purchase of animals, consumables, test kits etc.; The Institutional Research Theme of the University of Pretoria on Animal Zoonotic Disease and the Third Framework Agreement Programme (FA3-III) through the Institute of Tropical Medicine, Antwerp, Belgium to VR & AL (https://www.itg.be), and IRT funding was used towards all experimental costs, including purchase of animals, consumables, test kits etc.; MINECO and FEDER Plan Nacional (grant AGL2014-56305) to CG (http://www.idi.mineco.gob.es/portal/site/MICINN/menuitem.26172fcf4eb029fa6ec7da6901432ea0/?vgnextoid=4d494d861a299210VgnVCM1000001d04140aRCRD), and MINECO & FEDER funding was used towards coordination among IREC members; and the Erasmus Mundus Action 2 EUROSA scholarship to EvdH (https://www.uantwerpen.be/en/projects/eurosa/), and EUROSA funding was used towards the stipendium of EvdH.

The great atherogenic potential of oxidized low- density lipoprotein has been widely described in the literature. The objective ofthisstudy was to investigate whether the state of oxidized low-density lipoprotein in human plasma mea- sured by the Z-scan technique has an association with differ- ent cardiometabolic biomarkers. Total cholesterol, high- density lipoprotein cholesterol, triacylglycerols, apolipopro- tein A-I and apolipoprotein B, paraoxonase-1, and glucose were analyzed using standard commercial kits, and low- density lipoprotein cholesterol was estimated using the Friedewald equation. A sandwichenzyme-linked immunosor- bent assay was used to detect electronegative low-density li- poprotein. Low-density lipoprotein and high-density lipopro- tein sizes were determined by Lipoprint ® system. The Z-scan technique was usedto measure the non-linear optical response oflow-densitylipoproteinsolution.Principalcomponentanal- ysis and correlations were used respectively to resize the data from the sample and test association between the θ parameter, measured with the Z-scan technique, and the principal com- ponent. A total of 63 individuals, from both sexes, with mean age 52 years (±11), being overweight and having high levels of total cholesterol and low levels of high-density lipoprotein cholesterol, were enrolled in this study. A positive correlation betweenthe θparameter and moreanti-atherogenicpattern for cardiometabolic biomarkers together with a negative correla- tion for an atherogenic pattern was found. Regarding the pa- rameters related with an atherogenic low-density lipoprotein profile, the θ parameter was negatively correlated with a more atherogenic pattern. By using Z-scan measurements, we were able to find an association between oxidized low-density lipo- protein state and multiple cardiometabolic biomarkers in sam- ples from individuals with different cardiovascular risk factors.

The objective of this work was to study the effect of different hosts and temperatures on Diaphorina citri Kuwayama (Hem., Psyllidae) biology. Citrus limonia (Rangpur lime), Murraya paniculata (orange jessamine) and Citrus sunki (Sunki mandarin) were used as hosts. Measurements included duration and viability of the egg and nymphal stages, sex ratio, fecundity and longevity. In order to verify the effect of temperature on D. citri biology, the duration and viability of its developmental stages and biological cycle were compared at seven different temperature conditions. Durations of the embryonic and nymphal stages were similar for Rangpur lime, orange jessamine and mandarin. However, the nymphal viability obtained on mandarin was lower than the values obtained on the other hosts. Fecundity was higher on orange jessamine and, in all instances, females had greater longevity than males. A fixed number of instars (five) was obtained on the three hosts. Rangpur lime and orange jessamine provided better D. citri development when compared with mandarin. Duration of the egg and nymphal stages varied from 2.6 to 7.7 and from 9.4 to 35.8 days, respectively, at temperatures from 18 to 32� C. Egg viability was higher than 81.6% at the temperature range studied (18-32� C); nymphal viability was higher than 70% at the 18-30� C range, differing from viability at 32� C, which was dramatically reduced. The D. citri lower temperature development threshold (TT) and thermal constant (K) values for the egg, nymphal and biological cycle (egg-adult) stages were 12.0� C and 52.6 Degree-Day (DD); 13.9� C and 156.9 DD; and 13.5� C and 210.9 DD respectively.

Purpose: Averrhoa carambola L. has a long history of its use in traditional medicine for the treatment of various diseases such as diabetes, intermittent fevers, intestinal worms, scabies and inflammatory conditions. The fresh fruit of A. carambola is very popular in Sri Lanka for its glucose-lowering effect and is taken as good remedy for diabetes. The present study was carried out to isolate and characterize anti-hyperglycemic principles of the fruit of A. carambola to obtain additional scientific evidence for its usage in traditional medicine in the management of diabetes. Methods: The dried and powdered fruits of A. carambola were successively extracted with n-hexane and CH2Cl2 using soxhlet apparatus. The CH2Cl2 extract was subjected to silica gel column chromatography, eluting in a stepwise gradient with hexane and ethyl acetate (EtOAc) mixtures. The fraction eluted with hexane:EtOAc (60:40) yielded a mixture which with preparative thin-layer chromatography afforded a pure compound designated as JW-AC-3. The structure of JW-AC-3 was elucidated on the basis of its Ultra- violet spectroscopy (UV), Infra-Red spectroscopy (IR), Mass Spectroscopy (MS), and Nuclear Magnetic Resonance spectroscopy (NMR) including Distortionless Enhancement Polarization Transfer (DEPT), Homonuclear Shift-Corrilation Spectroscopy (COSY), Nuclear Overhauser Spectroscopy (NOESY), Hetero Multiple Bond Connectivities (HMBC) and Heteronuclear Multiple-Quantum Coherence (HMQC) experiments and direct comparison with reported data. Results: JW-AC-3 was identified as scopoletin by direct comparison of its spectral data with reported data of scopoletin. To our knowledge, scopoletin has not been reported from A. carambola. Scopoletin has been shown to possess significant anti-hyperglycemic activity by previous workers. The results of this study clearly indicate that the majority of the anti-hyperglycemic activity of the fruits of A. carambola could be attributed to scopoletin. The results also provide for the use of scopoletin as a bio-marker in the standardization and quality control of products derived from the fruits of A. carambola . Conclusion: The results of this study demonstrate potential use of scopoletin in the development of drugs of natural origin in the treatment of diabetes. The results could also be used in the rationalization of ethnomedical use of the fruits of A. carambola .

Bergenin was isolated as the major active constituent from the leaves of Flueggea leucopyrus Willd. The result of this study also provides for the use of F.leucopyrus as a novel source for the commercial supply of bergenin to the global market as bergenin is a medicinal agent required for the production of various pharmaceutical formulations used in the treatment of a variety of disorders.

Summary Early pregnancy factor (EPF) appears to be a protein complex that depends on pregnancy. It has been detected in the serum of several animal species using the Rosette Inhibition Test. In the present study 30 adult, Mangalarga mares with normal reproductive activity were divided into two groups: an experimental group which were mated and/or inseminated, and an unmated control group. In the experimental group, blood samples were taken on the 7th, 14th and 21st days after ovulation. In the control group, blood samples were collected from seven mares in oestrus and eight in dioestrus. In the control group the Rosette Inhibition Titre (RIT) ranged from 3 to 8 × 103, whereas in the experimental group a RIT higher than 8 × 103 was considered an indication of the presence of EPF. Serum progesterone concentrations on Day 7 were equal to or greater than those measured in the mated mares on Day 14 or Day 21.

On-site bioassays were conducted at the furthest upstream pulp mill on the Fraser River in British Columbia. Uncontaminated river water was used to dilute treated effluent as discharged from the final diffuser pond. A single cohort of juvenile (8-10gm) chinook salmon ( Oncorhynchus tshawytscha ) was divided into an hypoxic group receiving 65% oxygen saturated water and a normoxic group receiving ambient 88% oxygen saturated water. Both groups were exposed over a period of 30 days to effluent concentrations of 2%, 4%, 8%, and 16%, while the controls received uncontaminated river water. This range of concentrations spanned those encountered by wild juvenile salmon overwintering in the upper Fraser River mainstem. The blood when analyzed by flow cytometry showed significant concentration-dependent clastogenic damage in both the normoxic and hypoxic groups starting at the 4% concentration. A concentration-response curve was determined from the hypoxic data set. Genetic implications of mutagenic damage to natural populations of chinook salmon are discussed along with the utility of the flow cytometer in detecting genotoxic damage.

It has been suggested that an inhibitory ECM containing chondroitin-6- sulfate proteoglycan (C-6S-PG) and tenascin (TN), which appears homogeneously in the core of the OB following afferent fiber arrival, helps position ingrowing olfactory axons in the prospective glomerular layer (GL) (Gonzalez and Silver, 1992; Gonzalez et al., 1993). Later, a similar ECM associated with astrocytes envelopes axonal glomeruli in rings, suggesting that axons may control the precise ECM patterning. The question remains whether formation of the matrix ring pattern around each axonal glomerulus is an intrinsic property of the matrix- producing cells or a response to developing axons. To determine if the organization of glial associated matrix in the OB was dependent on the presence of axons, we studied the effect of unilateral injection of a neurotoxin into the olfactory epithelium of postnatal rats. Using olfactory marker protein (OMP), beta-tubulin (TUJ1) antibodies, and Nissl staining, we found that at 5 and 10 d following neurotoxin administration the number of glomeruli decreased by an average of 77.0% in the injected side. At the same time, we observed that the TN/C-6S-PG rings and periglomerular cells were present only around the remaining small number of glomeruli. Elsewhere, ECM expression and the periglomerular cell configuration were more disorganized in the GL. The pattern of glial fibrillary acidic protein (GFAP) did not change significantly. We found that OMP staining, beta-tubulin immunoreactivity, and periglomerular cells reformed in a glomerular- like pattern as the olfactory axons reformed by 20 d. As the glomeruli- shaped collection of axon terminals reappeared, TN/C-6S-PG immunoreactivity also reoccurred in rings around the new axon bundles. Again, at this later stage, the expression of GFAP was similar in both sides. In our previous study (Gonzalez et al., 1993), we suggested that the initial gross positioning of glomeruli may be controlled by the overall positioning of TN/C-6S-PG. In the present study, we suggest that the formation of TN/C-6S-PG in the precise ring pattern around glomeruli appears to be dependent upon the presence of bundled olfactory axons. Various mechanisms are discussed that may explain the dynamic change in ECM expression that occurs inside the glomerulus after the neurotoxin treatment.

Background Physical exercise in healthy individuals leads to an acute inflammatory-cell response in skeletal muscles. In COPD, quadriceps dysfunction is an important systemic manifestation that can be offset by exercise training. However, the nature of the muscle inflammatory response in these patients to acute and chronic exercise remains unknown. We therefore measured inflammatory cell infiltration in the quadriceps of COPD patients and healthy controls, in response to a programme of lower-limb resistance training. Methods 12 COPD patients (mean (SD) age 66.7(7.0) years, BMI 26.1(7.2) kg/m 2 , FEV 1 46.4 (20.5) % predicted, 10 males) and seven healthy controls (66.7 (5.1) years, BMI 27.7(2.4) kg/m 2 , FEV 1 103.4 (17.0) % predicted, five males) underwent 8 weeks of bilateral lower-limb, high-intensity resistance training, thrice weekly, on an isokinetic dynamometer (Cybex II Norm). Quadriceps muscle biopsies from the dominant thigh were obtained at baseline (T0), 24-h after the 1st exercise bout (T1), and 24-h after the last exercise session following 8-weeks training (T2). Glycol methacrylate-embedded muscle biopsies were analysed using immunohistochemistry. Inflammatory cells were identified using antibodies against neutrophil elastase (NE) and CD163 (macrophages). Dual energy x-ray Absorptiometry (DEXA) was used to determine thigh muscle mass and isometric quadriceps strength was measured on the cybex. Results At T0, neutrophils were not detected in any of the healthy controls, but were present in six out of 12 patients. A significant increase in neutrophil and macrophage counts in both patients and controls was seen at T1 (Abstract P142 Table 1). At T2, inflammatory cell counts in both groups were close to baseline values. Isometric quadriceps strength (COPD: pre vs post -134.0 (44.0) vs 151.8 (47.9) Nm, p=0.002; Healthy: pre vs post 153.4 (42.4) vs 169.0 (43.1) Nm, p=0.04] and thigh lean mass (COPD: pre vs post 4.4 (1.2) vs 4.7 (1.2) kg, p=0.003; Healthy: pre vs post 4.5 (0.6) vs 4.7 (0.7) kg, p=0.004] increased significantly after training in both groups. Conclusions Acute resistance exercise in COPD leads to an inflammatory-cell response in the quadriceps that is comparable to healthy controls. Regular training results in muscle adaptation characterised by a diminished inflammatory response to a bout of exercise.

Using the pyroantimoniate technique, a study was conducted at electron microscope level on the distribution of the calcium ion in the pineal glands of normal adult Sprague-Dawley rats with initial weights of 150-200 g subjected to a 12:12 light dark cycle and others under the same conditions were subjected to modifications in the noradrenergic signal, such as continuous illumination over 7 days, blinding by bilateral enucleation (7 or 90 days) before sacrifice and bilateral superior cervical gangliectomy at 21 days before sacrifice. All the animals were sacrificed by decapitation, half of them at midday and the other half at midnight. Abundant fine precipitations of calcium were found in the intercellular spaces of the pineal glands of the normal rats. By contrast, in the gangliectomized animals subjected to constant illumination and chronic binding these precipitations were few in number. Additionally, two types of pinealocytes were observed regarding the distribution and concentration of intracytoplasmic calcium in both the normal and experimentally manipulated animals. Type I correspond to the classic light pinealocytes, with an absence of intracytoplasmic precipitations, although in the normal and gangliectomized animals sacrificed at midnight it was possible to observe fine deposits inside the mitochondrial matrix. Type II correspond to the classic dark pinealocytes, with a dense cytoplasmic matrix and numerous deposits of intracytoplasmic and intranuclear calcium; these were never seen in the type I pinealocytes.

The somatic chromosome complement of Bubulcus ibis consists of six pairs of macrochromosomes, twenty-three pairs of microchromosomes and a pair of sex chromosomes. The Z-chromosome is comparable in size to autosome pairs 3 and 4, from which it is difficult to distinguish, and the W-chromosome is indistinguishable from the larger microchromosomes. The chromosome complement of one individual was found to deviate from the normal because of structural heterozygosity involving a member of chromosome pair 1 and a microchromosome.

The diploid chromosome number in the common house rat of Allahabad, Rattus rattus rufescens Gray., is 42 with XX/XY sex determining mechanism. The autosomes consists of 13 pairs of telocentrics (T or t) and 7 pairs of metacentrics (M or m). X is a medium-size, telocentric (T), ranking 5th or 6th in the order of length, in the complement, and constitutes 5.55 % of the total haploid length. Y is variable in size. Several karyotypic anomalies, such as the presence of an unpaired, large Sin or T chromosome, heteromorphic pair 1, close association of one of the homologues each of pairs 7 and 4, 7 and 12 and pair I and a large telocentric chromo-some, are reported.

SUMMARYThe behaviour of chromosomes during the spermatogenesis of the scorpion Palamnaeus longimanus has been studied. The following facts have been noticed: 1. The spermatogonial metaphase shows 64 chromosomes, and so does the oogonial metaphase.2. The male is the heterogametic sex, possessing X- and Y- chromosome.3. All the specimens of this species examined have been found to be reciprocal translocation heterozygotes for two pairs of chromosomes, which may, for example, be designated as AB, BC, CD, DA. The behaviour of these chromosomes is subject to variation. Mostly they pair to produce, at the first metaphase, two bivalents, or a single quadrivalent; sometimes pairing fails altogether or partly, resulting in the appearance of four univalents or one bivalent and two univalents. Hence the total number of chromosomal bodies at the first metaphase fluctuates between 31 and 34. This fluctuation is due at times to the dissociation of other bivalents.4. The bivalents lack chiasmata.5. The first anaphase po...

The development of neuromasts was studied in two species of teleosts, Cirrhina mrigala Ham. Buch. and Ophicephalus punctatus Bloch, and the findings are presented and discussed. It has been discovered, in the course of this investigation, that a neuromast arises by a process of invagination — a fact hitherto not reported. The occurrence of the dorsal lateral line and the accessory lateral line of the trunk, in addition to the main lateral line, has also been reported for the first time for teleosts, although known for other bony fishes.

SUMMARY1. The present work deals with the structure, behaviour and number of chromosomes of five fresh-water teleosts.2. The diploid and haploid chromosome numbers in these fishes are stated below: 1. Nandus nandus48242. Anabas testudineus48243. Colisa fasciatus48244. Glossogobius giuris43235. Rhynchobdella aculeata4824All the chromosomes in each species at spermatogonial metaphases are acrocentric, rod-shaped elements, and appear at meiotic metaphases as rounded in shape.3. No diplotene stage has been observed in any of the fish and, therefore, existence of chiasmata cannot be ascertained.4. Chromosome numbers and forms in related fished have been discussed in relation to taxonomy.5. No sex chromosomes are identifiable in any of the fishes studied, at any stage.

SUMMARY1. Specimens of Chrotogonus incertus were exposed to 400 r, 500 r and 800 r and spermatocytes were examined twentyfour hours subsequent to exposure.2. Exposure to 800 r caused aggregation of chromosomes into large vacuolated masses, which were inequally divided between the secondary spermatocytes. Another effect was extreme fragmentation of chromosomes.3. Exposure to 400 r and 500 r caused comparatively moderate aberrations—lagging and incomplete breaks.

The structure and behaviour of the chromosomes of male germ cells ofChrotogonus incertus have been studied. At the spermatogonial metaphase nine pairs of autosomes and one X-chromosome are observed.

1. The lampbrush chromosomes of Rana cyanophlyctis have been studied by phase-contrast microscopy.2. In the fully-formed lampbrush chromosomes, the loops are large and the chromomeres so small as to be barely noticeable. In the post-lampbrush stage chromosomes, the chromomeres are more massive and fewer, whereas there are no lateral loops.3. Chromonema is always single, but its duality is perhaps indicated by two lateral chromomeres at some positions on the chromonema.4. Loop bridges have been encountered in some preparations. These arise through accidents of manipulation.5. The chromomeres react positively to DNA tests, but not the loops and the chromonemata. The loops contain RNA, not found in the rest of the chromosomes.6. Nucleoli appear to arise chiefly on a middle-size chromosome.7. The nature of chromomeres, chiasmata and the origin of loops are discussed.

SUMMARYThe somatic chromosome complement of Clarius batrachus consists of (1) a pair of large metacentrics, (2) two pairs of small metacentrics, (3) 21 pairs of medium-size telocentrics and (4) two pairs of very small telocentrics.

The somatic chromosomes of seven species of Passeriformes belonging to six families have been studied. The diploid chromosome number varies from 68-80. These species possess each seven pairs of macrochromosomes, except Acridotheres tristis and Acridotheres ginginianus, which possess six pairs of macrochromosomes including the ZZ chromosomes.An identifiable W-chromosome has been found in all these species. It is either intermediate in size between macro- and microchromosomes or is of the size of the large microchromosomes. It has been found to be m, sm or T type chromosome.

THE sex-determining mechanism in dragon-flies consists, as a rule, of XO—XX chromosomes, and the X-chromosome in the spermatocyte divides equationally in the first, and passes undivided to one pole or the other in the second, meiotic division1–4. In this respect, the X-chromosome of the male germcell in the Odonata behaves as in the Heteroptera, in which group also it usually divides in the first meiotic division only. We find that the chromosomes of the damsel-fly, Ceriagrion coromandelianum, exhibit another important feature in respect of which they resemble the hemipteran chromosomes; that feature is the presence of Feulgen-positive interconnecting threads joining the chromosomes at the spermatogonial metaphase, and also at the first and second meiotic metaphase. Such interconnecting chromatic threads are known to exist in many hemipterans5–8. Associated with the presence of these threads is a high degree of heteropycnosis of autosome segments. At pachytene the paired chromosomes carry deeply stained, condensed knobs. These heterochromatic segments occur in all chromosome pairs. The interconnecting threads, in our opinion, are derived from the chromosome matrix. It is well known that the protein framework of the heterochromatic chromosomes (or chromosome segments) differs from that of euchromatic chromosomes9. It is likely that the nucleoprotein matrix synthesized by these segments is cohesive7 and that any fortuitous contact and subsequent separation between chromosomes result in the drawing out of the matrix substance into fine threads.

1. The spermatocytes and early spermatids of Vaginula possess a single Golgi element, consisting of an osmiophil externum and an osmiophobe internum. It is somewhat spherical, and is clearly observable in untreated living cells. In late spermatids the Golgi bodies break up into granules indistinguishable from mitochondria. 2. The acrosome is a small structure and arises without the intervention of the Golgi bodies. 3. The mitochondria are small osmiophil granules and rodlets. They remain scattered haphazard and do not give rise to Nebenkern or mitochondrial sheath of the middle piece. 4. The Golgi body does not consist of vacuome and ‘chondriome actif’. It stains neither with janus green B, nor with neutral red, but stains deeply with nile blue sulphate and methylene blue. 5. The mitochondria stain with janus green B, but not with nile blue sulphate. 6. Neutral-red bodies appear in the ground cytoplasm of the spermatocytes and spermatids. These are probably segregation droplets.

Der Verfasser macht am Golgikorper der mannlichen Geschlechtszellen vonVaginula eine Nachuntersuchung mit dem Phasenkontrastmikroskop und bestatigt, dass zwischen Golgikorperbildner und Akrosomgenese kein verwandtschaftlicher Zusammenhang besteht. Damit werden entgegenstehende Auffassungen vonNath widerlegt.

The meiotic chromosomes of the grasshopper, Chrotogonus incertus Bolivar (collected at Allahabad), assume typical ‘lampbrush’ appearance at pachytene, which condition persists to the end of diakinesis, but disappears completely at the first metaphase. This condition has been reported by Hsu1 in two different species of grasshoppers, and, judging from illustrations, it seems to have been observed in grasshoppers by other investigators also, even though they do not explicitly describe it2,3. Hsu1 has put forward an ingenious theory, based essentially on Goldschmidt's4 interpretation of the ‘lampbrush’ chromosome, which links up spiralization, heteropycnosis and the formation of ‘lampbrush’ processes. According to him, heterochromatinic heteropycnosis is not due, at least not entirely, to differential nucleic acid synthesis5, but to a difference in the capacity to undergo cyclic condensation. Negative heteropycnosis results from the fact that the intracellular conditions permit the growth of ‘lampbrush’ fibres on the X-chromosome, whereas the autosomes become compactly coiled. Positive heteropycnosis results when the conditions are reversed; the X-chromosome undergoes condensation, but not the autosomes, which, in consequence, show the ‘lampbrush’ fibres. Polymerization of the nucleoprotein, he claims, causes the chromosomes either to be coiled or show ‘lampbrush’ fibres. Now in the spermatocytes of Chrotogonus, late diakinetic chromosomes, although considerably condensed and consequently much shorter than the pachytene threads, are provided with ‘lampbrush’ fibres similar in size and shape to those of the earlier stages. So, condensation (spiralization) and ‘lampbrush’ formations can exist simultaneously in a chromosome. Further, the negatively heteropycnotic X-chromosome of early spermatogonial metaphase is not reported to possess ‘lampbrush’ fibres6,7. Also, in certain special cases, the rates of reproduction of euchromatin and heterochromatin have been found to be appreciably different8, so the differences between them may be more fundamental than Hsu's1 hypothesis assumes. In Chrotogonus the ‘lampbrush’ fibres spring from deeply stainable beads on the chromonemata. Such beads Serra9 claims to have resolved, on careful observation, into parts of a helicoidal coil of four chromonemata. He considers the ‘hairs’ of such chromosomes to be rod-like structures of nucleoplasm deposited on the chromonemata, and, in a lesser degree, the coils of chromonemata themselves. In view of the fact that the staining properties of the ‘lampbrush’ fibres are similar to those of the rest of the chromosome, and the almost unstaining nucleoplasm at these stages is markedly poor in nucleic acids, Serra's9 view is scarcely plausible. It is more in consonance with known facts to consider the ‘lampbrush’ bristles as arising from the peripheral nucleoprotein matrix surrounding the chromonemata10.

IN all the members of the family Nepidae (Hemiptera–Heteroptera) investigated so far from the above point of view, the diploid number of chromosomes in the male germ-cells has been reported to be 43–38 autosomes and 5 sex chromosomes1–4. Chickering's5 account of the chromosomes of Ranatra linearis, according to which the spermatogonia of this insect are of two kinds, one type possessing 40, and the other 48–50 chromosomes, has been shown to be erroneous by Steopoe2.

In den Spermatocyten vonSarcophaga ruficornis zeigen drei autosomale Bivalente scheinbare Chiasmata. Aus dem Vergleich der verschiedenen Stadien kann aber geschlossen werden, dass es sich nicht um Chiasmata, sondern um Beruhrung der Chromosomen an speziellen Punkten handelt. Auch chiasmaahnliche Bildungen homologer Chromosomen in der Mitose sind als Bindung homologer Chromatiden der geteilten Chromosomen aufzufassen.

Although there have been several studies on the use of immunohistochemical biomarkers of canine mammary tumors (CMTs), the results are difficult to compare. This article provides guidelines on the most useful immunohistochemical markers to standardize their use and understand how outcomes are measured, thus ensuring reproducibility of results. We have reviewed the biomarkers of canine mammary epithelial and myoepithelial cells and identified those biomarkers that are most useful and those biomarkers for invasion and lymph node micrometastatic disease. A 10% threshold for positive reaction for most of these markers is recommended. Guidelines on immunolabeling for HER2, estrogen receptors (ERs), and progesterone receptors (PRs) are provided along with the specific recommendations for interpretation of the results for each of these biomarkers in CMTs. Only 3+ HER2-positive tumors should be considered positive, as found in human breast cancer. The lack of any known response to adjuvant endocrine therapy of ER- and PR-positive CMTs prevents the use of the biological positive/negative threshold used in human breast cancer. Immunohistochemistry results of ER and PR in CMTs should be reported as the sum of the percentage of positive cells and the intensity of immunolabeling (Allred score). Incorporation of these recommendations in future studies, either prospective or retrospective, will provide a mechanism for the direct comparison of studies and will help to determine whether these biomarkers have prognostic significance. Finally, these biomarkers may ascertain the most appropriate treatment(s) for canine malignant mammary neoplasms.

The hammerhead ribozyme is a small catalytic RNA motif capable of endonucleolytic (self-) cleavage. It is composed of a catalytic core of conserved nucleotides flanked by three helices, two of which form essential tertiary interactions for fast self-scission under physiological conditions. Originally discovered in subviral plant pathogens, its presence in several eukaryotic genomes has been reported since. More recently, this catalytic RNA motif has been shown to reside in a large number of genomes. We review the different approaches in discovering these new hammerhead ribozyme sequences and discuss possible biological functions of the genomic motifs.