Cotransplantation of marginal mass allogeneic islets with 3D culture-derived adult human skin cells improves glycemia in diabetic mice

Islet transplantation represents a therapeutic option for type 1 diabetes (T1D). Long-term viability of transplanted islets requires improvement. Mesenchymal stromal cells (MSCs) have been proposed as adjuvants for islet transplantation facilitating grafting and functionality. Stem cell aggregation provides physiological interactions between cells and enhances the in situ concentration of modulators of inflammation and immunity. We established a hanging-drop culture of adult human skin fibroblast-like cells as spheroids, and skin spheroid-derived cells (SphCs) were characterized. We assessed the potential of SphCs in improving islet functionality by cotransplantation with a marginal mass of allogeneic islets in an experimental diabetic mouse model and characterized the secretome of SphCs by mass spectrometry-based proteomics. SphCs were characterized as multipotent progenitors and their coculture with anti-CD3 stimulated mouse splenocytes decreased CD4+ T cell proliferation with skewed cytokine secretion through an increase in the Th2/Th1 ratio profile. SphCs-conditioned media attenuated apoptosis of islets induced by cytokine challenge in vitro and importantly, intratesticular SphCs administration did not show tumorigenicity in immune-deficient mice. Moreover, SphCs improved glycemic control when cotransplanted with a marginal mass of allogeneic islets in a diabetic mouse model without pharmacological immunosuppression. SphCs' protein secretome differed from its paired skin fibroblast-like counterpart in containing 70% of up- and downregulated proteins and biological processes that overall positively influenced islets such as cytoprotection, cellular stress, metabolism, and survival. In summary, SphCs improved the performance of transplanted allogeneic islets in an experimental T1D model, without pharmacological immunosuppression. Future research is warranted to identify SphCs-secreted factors responsible for islets' endurance.

Effect of protein supplementation on yield and milk composition of F1 Holstein x Zebu cows kept in deferred pasture of brachiaria grass / [Efeito da suplementação de proteína na produção e na composição do leite de F1 vacas Holandês x Zebu mantidas em pasto diferido de capim-braquiária]

The objective of this study was to evaluate different supplementation strategies concentrated to F1 Holstein x Zebu lactating cows managed in deferred signal grass pasture on the yield and composition of milk and body weight gain. Thirty six F1 Holstein x Zebu cows with an average lactation period of 267 ± 10 days, mean body weight of 548 ± 19kg were used following a completely randomized design in a 4 x 5 factorial scheme, being four feeding strategies and five days under evaluation. The treatments consisted of four nutritional strategies: deferred pasture as a source of roughage without supplementation (PDSS); deferred pasture as a source of roughage with protein supplement offer (PDCS); deferred pasture supplemented with 15 kilos of corn silage (natural base) + 1,200 grams of protein supplement (PDSP) and corn silage (ad libitum) + 700 grams of protein supplement (SMP). There was no interaction (P> 0.05) between the nutritional plans and days under evaluation for any of the variables. It was found that cows fed SMP showed milk production 26.06% higher than the other nutritional plans (mean of 11.46kg/day). F1 Holstein/Zebu cows handled in deferred pasture in a traditional way supplemented with protein maintains milk yield.(AU)

Objetivou-se avaliar diferentes planos nutricionais para vacas F1 Holandês/Zebu mantidas em pasto diferido de capim-braquiária sobre a produção e a composição do leite e no ganho em peso corporal. Foram utilizadas 36 vacas F1 Holandês/Zebu com período médio de lactação de 267 ± 10 dias, peso corporal médio de 548 ± 19kg, seguindo o delineamento inteiramente ao acaso, em esquema fatorial 4 x 5, sendo quatro estratégias de alimentação e cinco dias em avaliação. Os tratamentos consistiram de quatro planos nutricionais: pasto diferida como fonte de forragem sem suplementação (PDSS); pastagem diferida como fonte de forragem com oferta de suplemento de proteico (PDCS); pasto diferido suplementado com 15 quilos de silagem de milho (base natural) + 1.200 gramas de suplemento proteico (PDSP) e silagem de milho (ad libitum) + 700 gramas de suplemento de proteína (SMP). Verificou-se que as vacas alimentadas com SMP apresentaram produção de leite 26,06% superior aos demais planos nutricionais (média de 11,46kg/dia). Vacas F1 Holandês/Zebu tratadas em pastagem diferida de maneira tradicional, suplementada com proteína, mantêm o rendimento de leite.(AU)

Effects of short-term feed restriction on the physiological parameters and metabolites of F1 Holstein x Zebu cows in different stages of lactation / [Efeitos da restrição alimentar de curto prazo sobre os parâmetros fisiológicos e metabólicos de vacas F1 Holstein x Zebu em diferentes estágios da lactação]

This study aimed to evaluate the effects of different nutritional plans on the productive, physiological and metabolic parameters of F1 ½ Holstein x ½ Zebu cows in different stages of lactation. Sixty lactating cows were allotted to a completely randomized 5 x 3 factorial design with five feed allowances and three lactation periods. The dry matter intake, milk yield and heart rate were reduced by 5.69kg, 2.41kg and 10.36 beats/min (morning) and 10.25 beats/min (afternoon) for each 1% feed restriction, respectively. There was no difference in the concentration of glucose, total protein, albumin, cholesterol and non-esterified fatty acids for cows subjected to different feed allowances, with means of 95.25, 7.98, 2.95, 121.68 and 0.45mg/dL, respectively. Feed restriction of up to 2.50% BW is a cost reduction strategy that does not alter milk yield, regardless of the stage of lactation.(AU)

Objetivou-se avaliar os efeitos de diferentes planos nutricionais sobre as características produtivas, fisiológicas e metabólicas de vacas F1 ½ Holandês x ½ Zebu. Foram utilizadas 60 vacas em lactação, seguindo-se o delineamento inteiramente ao acaso, em esquema fatorial 5 x 3, com cinco níveis de oferta de dieta e três períodos de lactação. À medida que se aumentou 1% na restrição da oferta da dieta, houve redução linear de 5,69kg no consumo de matéria seca pelos animais, 2,41kg na produção de leite, bem como de 10,36bat/min (manhã) e 10,25 bat/min (tarde) na frequência cardíaca dos animais. Não houve diferença para a concentração de glicose, proteínas totais, albumina, colesterol e NEFA com a restrição na oferta da dieta dos animais, sendo a média de 95,25, 7,98, 2,95, 121,68 e 0,45mg/dL, respectivamente. Recomenda-se a restrição de até 2,50% de peso corporal como estratégia de redução dos custos em todos os estágios em lactação, visando não alterar, economicamente, a produção de leite.(AU)

Caracterização fenotípica de vacas F1 Holandês x Zebu de diferentes bases maternas / [Phenotypic characterization of Holsteins x Zebu F1 cows from different maternal bases]

Caracterizaram-se fêmeas F1 Holandês x Zebu de diferentes bases maternas quanto às pelagens, despigmentações e características morfométricas. Foram utilizadas 266 fêmeas F1, progênies do cruzamento de 26 touros da raça Holandesa com fêmeas de composição genética zebuínas: Gir, Nelore, Guzonel, Nelogir. Foram aplicadas análise de distribuição de frequência para características qualitativas e medidas de dispersão e tendência central para características morfométricas, e as médias foram comparadas pelo teste de Tukey a 5% de probabilidade. Acima de 60,0% dos animais foram de pelagem preta. As vacas que tiveram origem na raça Gir apresentaram comprimento de cabeça 2,8cm maior (P<0,05) que as fêmeas da raça Nelore. O comprimento da orelha variou (P<0,05) conforme a base materna utilizada. As vacas com genes da raça Nelore são 5,0cm mais altas (P<0,05). O perímetro torácico foi a característica morfométrica que teve correlação fenotípica de elevada magnitude com o peso, acima de 0,70, para as fêmeas das bases maternas Gir, Nelore e Nelogir. A pelagem não é indicativa da base materna utilizada. As vacas F1 de base materna Gir tiveram estrutura corporal menor que as fêmeas que portam genes da raça Nelore.(AU)

F1 Holstein x Zebu females from different maternal bases were characterized regarding coat, depigmentation and morphometric characteristics. A total of 266 F1 female progenies from the crossbreeding of 26 Holstein bulls with females of Zebu genetic composition were used: Gir, Nellore, Guzonel, Nellogir. Frequency distribution analysis was applied for qualitative characteristics and dispersion measures and central tendency for morphometric characteristics, and means were compared by Tukey test at 5% probability. Over 60.0% of the animals had a black coat. The cows that originated from the Gir breed had a head length of 2.8cm (P<0.05) higher than the Nellore females. Ear length varied (P<0.05) according to the maternal base used. Cows with Nelore genes were 5.0cm taller (P<0.05). The thoracic perimeter was the morphometric characteristic that had a high magnitude phenotypic correlation with weight, above 0.70, for the females of the Gir, Nellore and Nellogir maternal bases. The coat is not indicative of the maternal base used. F1 Gir-based cows had a smaller body structure than females with Nellore genes.(AU)

Viabilidade econômica de vacas leiteiras F1 Holandês x Zebu com diferentes bases maternas e ordens de parto / Economic viability F1 Holstein x Zebu cows of dairy from different maternal bases and birth orders

Objetivou-se avaliar a viabilidade econômica de vacas F1 de Holandês x Zebu de diferentes bases maternas e ordens de parto. Foram utilizados dados do Campo Experimental da Epamig (Felixlândia-MG). Analisaram-se 406 lactações de vacas F1 H x Z de diferentes bases maternas. Estimaram-se a receita e os custos operacional efetivo, operacional total e custo total. As F1 Holandês x Gir tiveram médias de custos maiores (R$ 3582,30), seguidas pelas F1 Holandês x Nelogir (R$ 3448,06), e o menor foi das F1 Holandês x Nelore (R$ 3145,07). A receita total foi maior para as vacas F1 Holandês x Gir e F1 Holandês x Nelogir, com valores de R$ 4394,96 e R$ 4245,61, respectivamente, e a menor receita foi para F1 Holandês x Nelore, com valor de R$ 3976,12. O lucro foi de R$ 812, 65; R$ 797,54 e R$ 831,04 para as F1 filhas de base materna Gir, Nelogir e Nelore, respectivamente. Todas as bases maternas são viáveis economicamente e podem ser utilizadas no sistema de produção de leite e bezerros para venda. Todas as ordens de parto de Holandês x Zebu estudadas são viáveis economicamente.(AU)

The objective was to evaluate the economic viability of a milk production system of F1 Holstein x Zebu cows of different maternal bases and calving order. Lactations data of F1 Holtein x Zebu cows (n=406) of different maternal bases, from the experimental field of the EPAMIG at Felixlândia county (MG state, Brasil) were analyzed. Revenue, cost/effect of operation, total operating, and total cost have been estimated. F1 cows Holstein x Gir had averages of higher costs R$ 3582.30, followed by F1 cows Holstein x Nelogir R$ 3448.06; and the lowest was from cows F1 Holstein x Nellore with R$ 3145.07. The total revenue was higher for Holstein x Gir and Holstein x Nelogir with recipe values of R$ 4394.96 and R$ 4245.61 respectively, and the lowest value of R$ 3976.12 for Holstein x Nellore. The profits were R$812.65; R$ 797.54 and R$ 831.04 for animals of groups Holstein x Gir, Holstein x Nelogir and Holstein x Nellore respectively. All genetic groups are economically viable and can be used in the production of milk and calves for sale. All calving order of Holstein x Zebu studied are viable economically.(AU)

Effect of sugars on the association between cowpea vicilin (7S storage proteins) and fungal cells

Vicilins (7S storage proteins) found in various legume seeds have been previously shown to interfere with the germination of spores or conidia of phytopathogenic fungi and inhibit yeast growth and glucose stimulated acidification of the medium by yeast cells. In the present work vicilins from cowpea (Vigna unguiculata) seeds were added to the growth medium of Saccharomyces cerevisiae cells and Fusarium oxysporum conidia. Helix pomatia lectin, wheat germ agglutinin and Ulex europaeus lectin were used to identify differences in the binding of the vicilins to the surface of cells of S. cerevisiae and F. oxysporum treated with this protein. After the growth period, the material in suspension (yeast cells) was centrifuged and the final pellet was also treated with different sugar (glucose, sucrose, glucosamine, N-acetyl-glucosamine) concentrations and 0.1 M HCl for extraction of vicilins associated to chitinous structures present in yeast cells. Our results showed that vicilin sub-units were present in the different sugar extracts of yeast cells pretreated with the vicilins and these proteins were eluted by 0.5 M solutions of sugars in the following order of efficiency of elution: N-acetyl-glucosamine, sucrose/glucose and glucosamine.

Chitin-binding proteins from cowpea (Vigna unguiculata) seeds

Vicilins (7S storage proteins) from cowpea (Vigna unguiculata) and other legume seeds were shown to bind to chitin, to regenerated chitin (fully acetylated chitin) and to chitosan (deacetylated chitin). Adsorbed vicilins were desorbed from these matrices by acetic and hydrochloric acids and by highly polymerized soluble chitosan. Proteins such as the lectin of common bean (PHA), soybean trypsin inhibitor (Kunitz), a beta-1,3-glucanase from cowpea seeds, bovine pancreatic alpha-chymotrypsin, chicken ovalbumin, serum albumin and rabbit-gamma- globulin did not bind. The present result is the first description of vicilin binding to chitin but other proteins, such as wheat germ agglutinin (WGA), a lectin that contains the so called "chitin-binding domain", and a chitinase isolated from cowpea seeds, which are involved in the defense mechanisms of plants against insects and fungi, were also shown to bind to chitin as previously reported. The binding of vicilins to chitin is probably effected not through a "chitin-binding domain" because they do not share this sequence with the defense-related proteins cited above. We propose that this association of vicilins with chitin may be related to the effect of variant vicilins on the development of Callosobruchus maculatus (bruchid) in resistant cowpea seeds.