Structure, processing and midgut secretion of putative peritrophic membrane ancillary protein (PMAP) from Tenebrio molitor larvae

A cDNA coding for a Tenebrio molitor midgut protein named peritrophic membrane ancillary protein (PMAP) was cloned and sequenced. The complete cDNA codes for a protein of 595 amino acids with six insect-allergen-related-repeats that may be grouped in A (predicted globular)- and B (predicted nonglobular)-types forming an ABABAB structure. The PMAP-cDNA was expressed in Pichia pastoris and the recombinant protein (64 kDa) was purified to homogeneity and used to raise antibodies in rabbits. The specific antibody detected PMAP peptides (22 kDa) in the anterior and middle midgut tissue, luminal contents, peritrophic membrane and feces. These peptides derive from PMAP, as supported by mass spectrometry, and resemble those formed by the in vitro action of trypsin on recombinant PMAP. Both in vitro and in vivo PMAP processing seem to occur by attack of trypsin to susceptible bonds in the coils predicted to link AB pairs, thus releasing the putative functional AB structures. The AB-domain structure of PMAP is found in homologous proteins from several insect orders, except lepidopterans that have the apparently derived protein known as nitrile-specifier protein. Immunocytolocalization shows that PMAP is secreted by exocytosis and becomes entrapped in the glycocalyx, before being released into midgut contents. Circumstantial evidence suggests that PMAP-like proteins have a role in peritrophic membrane type 2 formation. (C) 2007 Elsevier Ltd. All rights reserved.

Sequencing of Spodoptera frugiperda midgut trehalases and demonstration of secretion of soluble trehalase by midgut columnar cells

Both soluble (SfTre1) and membrane-bound (SfTre2) trehalases occur along the midgut of Spodoptera frugiperda larvae. Released SfTre2 was purified as a 67 kDa protein. Its K(m) (1.6 mM) and thermal stability (half life 10 min at 62 degrees C) are different from the previously isolated soluble trehalase (K(m) = 0.47 mM; 100% stable at 62 degrees C). Two cDNAs coding for S. frugiperda trehalases have been cloned using primers based on consensus sequences of trehalases and having as templates a cDNA library prepared from total polyA-containing RNA extracted from midguts. One cDNA codes for a trehalase that has a predicted transmembrane sequence and was defined as SfTre2. The other, after being cloned and expressed, results in a recombinant trehalase with a K(m) value and thermal stability like those of native soluble trehalase. This enzyme was defined as SfTre1 and, after it was used to generate antibodies, it was immunolocalized at the secretory vesicles and at the glycocalyx of columnar cells. Escherichia coli trehalase 3D structure and sequence alignment with SfTre1 support a proposal regarding the residue modulating the pKa value of the proton donor.

Optimisation of solid-phase microextraction combined with gas chromatography–mass spectrometry based methodology to establish the global volatile signature in pulp and skin of Vitis vinifera L. grape varieties

The volatiles (VOCs) and semi-volatile organic compounds (SVOCs) responsible for aroma are mainly present in skin of grape varieties. Thus, the present investigation is directed towards the optimisation of a solvent free methodology based on headspace-solid-phase microextraction (HS-SPME) combined with gas chromatography–quadrupole mass spectrometry (GC–qMS) in order to establish the global volatile composition in pulp and skin of Bual and Bastardo Vitis vinifera L. varieties. A deep study on the extraction-influencing parameters was performed, and the best results, expressed as GC peak area, number of identified compounds and reproducibility, were obtained using 4 g of sample homogenised in 5 mL of ultra-pure Milli-Q water in a 20 mL glass vial with addition of 2 g of sodium chloride (NaCl). A divinylbenzene/carboxen/polydimethylsiloxane fibre was selected for extraction at 60 °C for 45 min under continuous stirring at 800 rpm. More than 100 VOCs and SVOCs, including 27 monoterpenoids, 27 sesquiterpenoids, 21 carbonyl compounds, 17 alcohols (from which 2 aromatics), 10 C13 norisoprenoids and 5 acids were identified. The results showed that, for both grape varieties, the levels and number of volatiles in skin were considerably higher than those observed in pulp. According to the data obtained by principal component analysis (PCA), the establishment of the global volatile signature of grape and the relationship between different part of grapes—pulp and skin, may be an useful tool to winemaker decision to define the vinification procedures that improves the organoleptic characteristics of the corresponding wines and consequently contributed to an economic valorization and consumer acceptance.

Effects of the basic fibroblast growth factor and its anti-factor in the healing and collagen maturation of infected skin wound

PURPOSE: The infection is one of the main factors that affect the physiological evolution of the surgical wounds. The aim of this work is to evaluate the effects of fibroblast growth factor (FGFâ) and anti-FGFâ in the healing, synthesis and maturation of collagen when topically used on infected skin wounds of rats. METHODS: An experimental study was perfomed in 60 male Wistar rats. All animals were divided in two groups (A and B). Each group was divided in three subgroups A1, B1; A2, B2 and A3, B3. After anesthesia with pentobarbital, two open squared wounds (1cm2), 4cm distant to each other, were done in the dorsal skin of all the rats. In group A (n=30) the wounds were contaminated with multibacterial standard solution, and in group B(n=30) the wounds were maintained sterile. These wounds were named F1 (for inflammation analysis) and F2 (for collagen study). The open wounds of A1 and B1 rats were topically treated with saline solution, A2 and B2 were treated with FGFâ and subgroups A3 and B3 were treated with FGFâ and anti-FGFâ. The rats were observed until complete epitelization of F2 wounds for determination of healing time and the expression of types I and III collagen, using Picro Sirius Red staining. Inflammatory reaction in F1 wounds was studied using hematoxilineosin staining. The three variable was measured by the Image Pro-Plus Média Cybernetics software. The statistical analysis was performed by ANOVA and Tukey test, considering p<0.05 as significant. RESULTS: It was observed that infection retarded significantly (p<0.05) the time of wound scarring and the topical application of FCFb reverted the inhibition of healing caused by bacteria. The inflammatory reaction was greater in the subgroup B2 than in B1 and A3, and the difference was significant (p<0.05). It was observed greater expression of type I collagen in all the subgroups treated with FCFb, when compared with the untreated subgroups. Type III collagen was significantly decreased in wounds of B3 rats, comparing to the other subgroups. CONCLUSIONS: The FCFb accelerated the healing of open infected wounds and contributed with maturation of collagen, enhancing the type I collagen density. The anti-FCFb antibody was able to attenuate the production of both type I and III collagen

Simvastatin improves the healing of infected skin wounds of rats

This study explores the potential of the simvastatin to ameliorate inflammation and infection in open infected skin wounds of rats. Methods: Fourteen Wistar rats weighing 285±12g were used. The study was done in a group whose open infected skin wounds were treated with topical application of sinvastatina microemulsion (SIM, n=7) and a second group with wounds treated with saline 0.9 % (SAL, n=7). A bacteriological exam of the wounds fluid for gram positive and gram negative bacteria, the tecidual expression of TNFá and IL-1â by imunohistochemical technique, and histological analysis by HE stain were performed. Results: The expression of TNFa could be clearly demonstrated in lower degree in skin wounds treated with simvastatin (668.6 ± 74.7 ìm2) than in saline (2120.0 ± 327.1 ìm2). In comparison, wound tissue from SIM group displayed leukocyte infiltration significantly lower than that observed in SAL group (p<0.05). Culture results of the samples taken from wound fluid on fourth post treatment day revealed wound infection in only one rat of group simvastatin (SIM), where Proteus mirabilis, Escherchia coli and Enterobacter sp were isolated. In the rats whose wounds were treated with saline (SAL), polymicrobial infection with more than 100,000 CFU/g was detected in all the wounds. Conclusion: In addition to its antiinflammatory properties, the protective effects of simvastatin in infected open skin wounds is able to reduce infection and probably has antibacterial action. The potential to treat these wounds with statins to ameliorate inflammation and infection is promising

Role of phospholipase A(2) and tyrosine kinase in Clostridium difficile toxin A-induced disruption of epithelial integrity, histologic inflammatory damage and intestinal secretion

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

A Catalytically Inactive Lys49 PLA2 Isoform from Bothrops jararacussu venom that Stimulates Insulin Secretion in Pancreatic Beta Cells

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Intraocular pressure and tear secretion in Saanen goats with different ages

Objetivou-se comparar os valores normais de pressão intraocular (PIO) e de produção lacrimal, através de tonometria de aplanação e do teste da lágrima de Schirmer-1 (TLS-1) em cabras da raça Saanen, com diferentes idades e momentos. Trinta e seis cabras da raça Saanen, livres de afecções oculares, foram agrupadas em três categorias etárias (n=12), compreendendo animais com 45, 180 e 549 dias de idade. O TLS-1 e a PIO foram aferidos sempre às 9:00 e às 19:00, durante três dias consecutivos. Os resultados foram avaliados estatisticamente (p<0,05). Relativamente ao horário do dia, a PIO foi significativamente menor às 19:00 (p<0,001) nos animais com 45 dias de idade; os valores do TLS-1 foram significativamente maiores às 19:00 (p=0,004) nas cabras com 549 dias de idade. Considerando-se a somatória dos três dias, ambos os parâmetros foram significativamente menores nos indivíduos com 45 dias de idade (p<0,001). A pressão intraocular e a produção lacrimal em cabras da raça Saanen se elevam de forma significativa até os 180 dias de idade.

Surgical treatment of gynaecomastia in a Saanen goat

Background: Gynaecomastia in male goats is characterized by abnormal development of the mammary gland. Enlarged udder may be observed cranially to the scrotum, which can occasionally reach the size of the testicles. The udder may carry functional glands and impair the animal's reproductive performance and welfare. The case of a successful surgical treatment of gynaecomastia in a high reproductive performance Saanen buck-goat is reported in the present study.Material, Methods & Results: The animal was admitted presenting significant augmentation of the mammary glands, which was clinically diagnosed as gynaecomastia. The male goat owned optimal phenotypic characteristics for the Saanen breed, which had been producing high performance descendents. The mammary glands had been impairing the goat's locomotion and sexual performance. Manual milking resulted in great amount of milk secretion. The animal presented anorexia and impaired sexual performance. After clinical and laboratorial evaluation, the animal was submitted to radical mastectomy. An elliptic skin incision was performed around each mammary gland. Subcuticular blunt dissection was accomplished to isolate the mammarian tissue from the abdominal muscular layer and the spermatic chord. The excised mass was sampled for histological assessment. Subcuticular layer and skin closure was carried in a routine fashion. Hygienization of the surgical wound was performed with 2,5% PVP-I solution for ten days. Additionally, an association of penicillin G benzathine and streptomycin, and fluxinin meglumine were also given. The surgical procedure was successfully accomplished without any peroperative complication. The excised mass was sampled for anatomic/histological assessment. Macroscopically, the left mammary gland presented 22 cm in length, 12 cm wide and 26 cm in diameter. The right gland presented 16 cm in length, 7 cm wide and 13,5 cm in diameter. The microscopic assessment revealed hyperplasia of the glandular ducts. No abnormalities resembling malignant mammary neoplasms or degeneration were observed. At the end of the treatment, the animal was completely recovered. The animal convalesced satisfactorily and surgical wound healed completely within the first 10 days post-op. The goat was not culled and returned to normal reproductive activity. Within 12 months of follow-up, the animal was able to produce high milk yield performance progenies.Discussion: This case report presented relevant aspects of the surgical management of gynaecomastia, especially to veterinary practitioners dealing with milk goats. Gynaecomastia is not as common as other reproductive disorders in domestic animals. In opposition to the findings of the present study, other trials revealed that gynaecomastia usually does not affect fertility, libido, ejaculate parameters and sexual performance of goats. However, it is important to consider that neoplasic disorders such as mammary adenocarcinoma may be present, even though these are rare complications. Last but not least, the decision making on mastectomy in the present study was crucial in order to reestablish the animal's welfare and its functionality in the farms reproduction program.

Detection of Leishmania (L.) chagasi in canine skin

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of the season of the year, truck type and location on truck on skin bruises and meat quality in pigs

The purpose of this study was to evaluate the effect of the season of the year (summer vs winter), type of truck (A: single decker vs B : double-decker) and pig location on the truck (front, middle, rear) on the incidence of skin bruising and pork quality variation. For this purpose, 2660 gilts of an average weight of 126.7 (+/- 6.6) kg originating from 19 different farms were used. No interaction between season of the year, type of truck and location on truck was observed. A higher number of bruises on the body at unloading and slaughter (P < 0.0001) and a higher number of bruises on the carcass (P < 0.01) were observed in winter. At unloading a higher number of bruises on the body and on the carcass after slaughter was observed in pigs transported on Truck A (P=0.004 and P=0.05). A higher, although not significant, number of bruises was found on the body of pigs transported in the rear compartment of both trucks. Higher paleness value was found in the longissimus and semimembranosus muscles in summer (P=0.0001) than in winter. Cold and heat stress have a negative influence on skin bruises and meat quality, respectively. Poor vehicle design increases the incidence of bruised carcasses without detracting from pork quality regardless of the climate conditions tested and location of the animal it] the truck. (c) 2006 Elsevier B.V. All rights reserved.

The field resistance of potato cultivars to foliar late blight and its relationship with foliage maturity type and skin type in Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Comparison of the properties of compacted and porous lamellar chitosan-xanthan membranes as dressings and scaffolds for the treatment of skin lesions

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)