Approximate expression for the ground-state energy of the two- and three-dimensional Hubbard model at arbitrary filling obtained from dimensional scaling.

We generalize the linear discrete dimensional scaling approach for the repulsive Hubbard model to obtain a nonlinear scaling relation that yields accurate approximations to the ground-state energy in both two and three dimensions, as judged by comparison to auxiliary-field quantum Monte Carlo (QMC) data. Predictions are made for the per-site ground-state energies in two and three dimensions for n (filling factor) and U (Coulomb interaction) values for which QMC data are currently unavailable.

Short report: evaluation of the potency and stability of a candidate vaccine against American cutaneous leishmaniasis.

Availability of a safe, immunogenic, and affordable vaccine would represent the best strategy for control of cutaneous leishmaniasis (CL). Stability in field conditions is a essential property for any candidate vaccine. The stability and immunogenicity of three different preparations (thimerosal-preserved, autoclaved, and lyophilized) of a killed Leishmania amazonensis vaccine were assessed using fresh products and after 12 months of storage at 4 degrees C. Autoclaving was associated with a time-dependent decrease in the immunogenicity of the vaccine, as measured by the leishmanin skin test and production of interferon-gamma. These findings are of importance in the decision of which preparation of candidate killed CL vaccines should move to phase III trials.

Purification of bovine pancreatic glucagon as a by-product of insulin production.

A process for purifying bovine pancreatic glucagon as a by-product of insulin production is described. The glucagon-containing supernatant from the alkaline crystallization of insulin was precipitated using ammonium sulfate and isoelectric precipitation. The isoelectric precipitate containing glucagon was then purified by ion-exchange chromatography on Q-Sepharose FF, gel filtration on Sephadex G-25 and ion-exchange chromatography on S-Sepharose FF. A pilot scale test was performed with a recovery of 87.6% and a purification factor of 8.78 for the first chromatographic step, a recovery of 75.1% and a purification factor of 3.90 for the second, and a recovery of 76.2% and a purification factor of 2.36 for the last one. The overall yield was 50%, a purification factor of 80.8 was obtained and the fraction containing active glucagon (suitable for pharmaceutical preparations) was 84% pure as analyzed by HPLC.

Microencapsulation and tissue engineering as an alternative treatment of diabetes.

In the 70's, pancreatic islet transplantation arose as an attractive alternative to restore normoglycemia; however, the scarcity of donors and difficulties with allotransplants, even under immunosuppressive treatment, greatly hampered the use of this alternative. Several materials and devices have been developed to circumvent the problem of islet rejection by the recipient, but, so far, none has proved to be totally effective. A major barrier to transpose is the highly organized islet architecture and its physical and chemical setting in the pancreatic parenchyma. In order to tackle this problem, we assembled a multidisciplinary team that has been working towards setting up the Human Pancreatic Islets Unit at the Chemistry Institute of the University of São Paulo, to collect and process pancreas from human donors, upon consent, in order to produce purified, viable and functional islets to be used in transplants. Collaboration with the private enterprise has allowed access to the latest developed biomaterials for islet encapsulation and immunoisolation. Reasoning that the natural islet microenvironment should be mimicked for optimum viability and function, we set out to isolate extracellular matrix components from human pancreas, not only for analytical purposes, but also to be used as supplementary components of encapsulating materials. A protocol was designed to routinely culture different pancreatic tissues (islets, parenchyma and ducts) in the presence of several pancreatic extracellular matrix components and peptide growth factors to enrich the beta cell population in vitro before transplantation into patients. In addition to representing a therapeutic promise, this initiative is an example of productive partnership between the medical and scientific sectors of the university and private enterprises.

Suplementação de novilhos Nelore sob pastejo, no período de transição águas-seca / Supplementation of Nelore steers under grazing during the rainy/dry transition period

Avaliou-se o efeito da suplementação proteica sobre o consumo e o desempenho de novilhos recriados em pastagens de capim-piatã (Brachiaria brizantha cv. Piatã), durante o período de transição águas-seca. Utilizaram-se 20 novilhos Nelore, com peso médio inicial de 260kg, distribuídos ao acaso, em esquema de parcelas subdivididas. Os suplementos foram: sal mineral com ureia (controle) - ofertado ad libitum; sal proteinado - ofertado a 0,2% do peso vivo; suplemento proteico-energético - ofertado a 0,3% do peso vivo; e suplemento proteico-energético - ofertado a 0,5% do peso vivo. A suplementação teve efeito aditivo sobre o consumo de matéria seca total. O consumo médio diário dos suplementos foi de 0,167; 0,597; 0,865 e 1,469kg/animal, sendo observado ganho médio diário de 0,686; 0,761; 0,719 e 0,850kg/animal para os tratamentos controle e suplementados com 0,2; 0,3 e 0,5% do peso vivo, respectivamente. Verificou-se que as estratégias de suplementação avaliadas foram economicamente viáveis e proporcionaram desempenho semelhante sob condições de elevada oferta de forragem, sendo recomendado iniciar a suplementação proteica no período de transição águas-seca.

The effects of proteic supplementation on performance and forage dry matter intake of steers grazing on Brachiaria grass (Brachiaria brizantha cv. Piatã) during the rainy/dry transition period were evaluated. Twenty Nelore steers with an average body weight (BW) of 260kg, were assigned in a split plot design. The treatments were: mineral salt plus urea offered ad libitum (control); proteic supplement offered at 0.2% BW, proteic-energetic supplement offered at 0.3% BW and proteic-energetic supplement offered at 0.5% BW. The supplementation increased total dry matter intake. The average intake of supplements was 0.167; 0.597; 0.865 and 1.469kg/animal/day and the average daily gain (ADG) was 0.686; 0.761; 0.719 and 0.850kg/animal/day, for control, 0.2, 0.3 and 0.5% BW, respectively. It was verified that all strategies of supplementation are economically viable and similar in performance, under high herbage availability conditions. It is suggested that proteic supplementation begin during the rainy/dry transition period, for better animal performance and economic results.

Antiulcerogenic activity of a crude hydroalcoholic extract and coumarin isolated from Mikania laevigata Schultz Bip.

The leaves of Mikania (Asteraceae) species are used in folk medicine as antispasmodic, antiulcerogenic and antirheumatic agents. Phytochemical screening of the crude hydroalcoholic 70% extract (CHE) of Mikania laevigata Shultz Bip. revealed coumarins, terpenes and organic acids. Antiulcerogenic activity of CHE was evaluated, employing different experimental models in rats, to discern the pharmacological mechanism of action. Both the antisecretory and the cytoprotection hypothesis were evaluated. The crude hydroalcoholic extract (1000 mg/kg body wt., vo) decreased the ulcerative lesion index produced by indomethacin, ethanol, stress and reserpine in rats by 85%, 93%, 82% and 50%, respectively. In the pyloric ligation model, a decrease of hydrogenionic concentration (53%) was observed, suggesting that the pharmacological mechanism has a relationship to antisecretory activity. The antisecretory mechanisms of CHE and the coumarin isolated from M. laevigata were confirmed by acid hypersecretion induced by histamine, pentagastrin and bethanechol. Duodenal administration of CHE (1000 mg/kg body wt.) and coumarin (100 mg/kg body wt.) inhibited only the gastric acid secretion produced by bethanecol. These results suggest that both CHE and coumarin may influence the secretion control mediated by the parasympathetic system.

Protective immunity using recombinant human IL-12 and alum as adjuvants in a primate model of cutaneous leishmaniasis.

Protection from cutaneous leishmaniasis, a chronic ulcerating skin lesion affecting millions, has been achieved historically using live virulent preparations of the parasite. Killed or recombinant Ags that could be safer as vaccines generally require an adjuvant for induction of a strong Th1 response in murine models. Murine rIL-12 as an adjuvant with soluble Leishmania Ag has been shown to protect susceptible mice. We used 48 rhesus macaques to assess the safety, immunogenicity, and efficacy of a vaccine combining heat-killed Leishmania amazonensis with human rIL-12 (rhIL-12) and alum (aluminum hydroxide gel) as adjuvants. The single s.c. vaccination was found to be safe and immunogenic, although a small transient s.c. nodule developed at the site. Groups receiving rhIL-12 had an augmented in vitro Ag-specific IFN-gamma response after vaccination, as well as increased production of IgG. No increase in IL-4 or IL-10 was found in cell culture supernatants from either control or experimental groups. Delayed hypersensitivity reactions were not predictive of protection. Intradermal forehead challenge infection with 107 metacyclic L. amazonensis promastigotes at 4 wk demonstrated protective immunity in all 12 monkeys receiving 2 microgram rhIL-12 with alum and Ag. Partial efficacy was seen with lower doses of rhIL-12 and in groups lacking either adjuvant. Thus, a single dose vaccine with killed Ag using rhIL-12 and alum as adjuvants was safe and fully effective in this primate model of cutaneous leishmaniasis. This study extends the murine data to primates, and provides a basis for further human trials.

Recombinant human insulin. VIII. Isolation of fusion protein--S-sulfonate, biotechnological precursor of human insulin, from the biomass of transformed Escherichia coli cells.

Various methods have been investigated for the isolation and purification of fusion proteins of precursors of human insulin in the form of S-sulfonates, from the biomass of transformed Escherichia coli cells. Fusion proteins were prepared with different sizes and structures of the leader peptide and the poly-His position (inserted for purification by metal chelate affinity chromatography). The fusion proteins contained an IgG-binding B domain of protein A from Staphylococcus aureus at the N-terminus and an Arg residue between the leader peptide of the molecule and the proinsulin sequence, for trypsin cleavage of the leader peptide. Six residues of Cys in proinsulin allow the chemical modification of the protein as a (Cys-S-SO(-)(3))(6) derivative (S-sulfonate), which increases its polyelectrolytic properties and improves the efficiency of its isolation. Various methods of oxidative sulfitolysis were compared with catalysis by sodium tetrathionate or cystine and Cu2+ or Ni2+ ions. An optimum scheme for the isolation and purification of S-sulfonated fusion proteins was developed by the combination of metal-chelating affinity and ion-exchange chromatography. Highly purified (95%) S-sulfonated fusion protein was recovered which was 85% of the fusion protein contained in the biomass of E. coli cells. Folding of fusion protein S-sulfonate occurred with high yield (up to 90-95%). We found that the fusion protein-S-sulfonate has proinsulin-like secondary structure. This structure causes highly efficient fusion protein folding.

Osseointegração clínica-radiológica do compósito hidroxiapatita-lignina entre implante metálico e tecido ósseo em coelho / Clinical-radiographic osseointegration of hydroxyapatite-lignin composite between metallic implant and bone tissue in rabbit

Para avaliar o compósito hidroxiapatita-lignina na osseointegração entre implante metálico e o tecido ósseo, foram utilizados 20 coelhos adultos, nos quais foi realizada uma falha óssea na face lateral proximal de ambas as tíbias. Na tíbia esquerda, introduziu-se, no canal medular, um pino intramedular de Schanz revestido com o compósito em sua parte rosqueada, após o preenchimento daquele com 1000mg do compósito. A falha cortical foi preenchida com o compósito. O mesmo foi feito na tíbia direita, porém sem a utilização do compósito, servindo como controle. A avaliação clínica baseou-se na deiscência, claudicação, sensibilidade dolorosa e circunferência tibial. Foram realizadas radiografias imediatamente após a cirurgia e aos oito, 30, 60, 90 e 120 dias do pós-operatório. A maioria dos animais apresentou evolução clínica normal. Nas radiografias do grupo tratado, houve decréscimo da radiopacidade no defeito e no espaço medular até tornar-se semelhante à do osso circunvizinho, quadro inverso ao do grupo-controle. Conclui-se que o compósito hidroxiapatita-lignina não mostrou indícios clínicos de rejeição e que o tecido visualizado na altura da falha óssea e ao redor do pino intramedular tinha radiopacidade semelhante à do osso circunvizinho, o que sugere que o material promoveu a integração com o tecido ósseo.

This study evaluated a sinthetic hydroxyapatite-lignin composite for osseoingration between metallic implant and bone tissue. Twenty New Zealand rabbits were used. A defect was made a the proximal region of lateral surfaces of both tibias. An intramedullary pin was inserted in the medullary cavity of the left tibia after filling the cavity with 1,000mg of the composite and covering the thread part of the pin with the composite. The same procedure was made in control tibias without the composite. Clinical evaluation was based on inflammatory reaction, dehiscence, lameness, pain, and tibial circumference. Radiographs were made immediately after surgery and then on days eight, 30, 60, 90, and 120. Most of the animals presented normal clinical progression. Radiographies of treated group showed decrease in the defect and medullary cavity radiopacity to the same pattern of the surrounding bone, while the contrary happened in control group; but, in this group, the medullary cavity remained radioluscent at the last observation date. It can be concluded that the hydroxyapatite-lignin composite did not show clinical signs of rejection and that radiopacity of the tissue in the bone defect and surrounding the pin was similar to bone radiopacity, suggesting that the material promoted osseointegration.

Purification of bovine pancreatic glucagon as a by-product of insulin production

A process for purifying bovine pancreatic glucagon as a by-product of insulin production is described. The glucagon-containing supernatant from the alkaline crystallization of insulin was precipitated using ammonium sulfate and isoelectric precipitation. The isoelectric precipitate containing glucagon was then purified by ion-exchange chromatography on Q-Sepharose FF, gel filtration on Sephadex G-25 and ion-exchange chromatography on S-Sepharose FF. A pilot scale test was performed with a recovery of 87.6 percent and a purification factor of 8.78 for the first chromatographic step, a recovery of 75.1 percent and a purification factor of 3.90 for the second, and a recovery of 76.2 percent and a purification factor of 2.36 for the last one. The overall yield was 50 percent, a purification factor of 80.8 was obtained and the fraction containing active glucagon (suitable for pharmaceutical preparations) was 84 percent pure as analyzed by HPLC.

Microencapsulation and tissue engineering as an alternative treatment of diabetes

In the 70's, pancreatic islet transplantation arose as an attractive alternative to restore normoglycemia; however, the scarcity of donors and difficulties with allotransplants, even under immunosuppressive treatment, greatly hampered the use of this alternative. Several materials and devices have been developed to circumvent the problem of islet rejection by the recipient, but, so far, none has proved to be totally effective. A major barrier to transpose is the highly organized islet architecture and its physical and chemical setting in the pancreatic parenchyma. In order to tackle this problem, we assembled a multidisciplinary team that has been working towards setting up the Human Pancreatic Islets Unit at the Chemistry Institute of the University of São Paulo, to collect and process pancreas from human donors, upon consent, in order to produce purified, viable and functional islets to be used in transplants. Collaboration with the private enterprise has allowed access to the latest developed biomaterials for islet encapsulation and immunoisolation. Reasoning that the natural islet microenvironment should be mimicked for optimum viability and function, we set out to isolate extracellular matrix components from human pancreas, not only for analytical purposes, but also to be used as supplementary components of encapsulating materials. A protocol was designed to routinely culture different pancreatic tissues (islets, parenchyma and ducts) in the presence of several pancreatic extracellular matrix components and peptide growth factors to enrich the beta cell population in vitro before transplantation into patients. In addition to representing a therapeutic promise, this initiative is an example of productive partnership between the medical and scientific sectors of the university and private enterprises.