COORDINATE EXPRESSION OF CYTOKERATIN-7 AND CYTOKERATIN-20 DEFINES UNIQUE SUBSETS OF CARCINOMAS

We tested the hypothesis that the coordinate expression of cytokeratin 7 (CK 7) and cytokeratin 20 (CK 20) could distinguish among carcinomas arising from different primary sites. A total of 384 cases of carcinomas primary to various organs, as well as 16 cases of malignant mesothelioma, were evaluated using commercially available monoclonal antibodies and an avidin-biotin immunoperoxidase technique. The subset of tumors strongly expressing both CK 7 and CK 20 included virtually all bladder transitional cell carcinomas and the majority of pancreatic adenocarcinomas; the tumors negative for both CK 7 and CK 20 were largely restricted to hepatocellular, prostate, and renal cell carcinomas in addition to squamous cell and neuroendocrine carcinomas of lung. The CK 7-/CK 20+ immunophenotype, however, was highly characteristic of adenocarcinomas of colorectal origin, whereas CK 7+/CK 20- immunophenotype was typically seen in the vast majority of carcinomas arising from other sites, including ovary, endometrium, breast, and lung, as well as malignant mesothelioma. Gastric carcinomas were the most heterogeneous subgroup with respect to CK 7/CK 20 immunophenotype. In the subset of mucinous tumors, striking immunophenotypic differences were noted among those primary to the breast (CK 7+/CK 20-), gastrointestinal tract (CK 7-/CK 20+), and ovary (CK 7+/CK 20+). In all cases investigated, this CK immunophenotype was invariant in metastatic vs. primary tumors. It is concluded that, in the appropriate clinical setting, the CK 7/CK 20 immunophenotype of carcinomas is a valuable diagnostic marker in the determination of primary site of origin.

Genetic parameters of milk, fat, and protein yields in the first three lactations, using an animal model and restricted maximum likelihood

Milk, fat, and protein yields of Holstein cows from the States of New York and California in the United States were used to estimate (co)variances among yields in the first three lactations, using an animal model and a derivative-free restricted maximum likelihood (REML) algorithm, and to verify if yields in different lactations are the same trait. The data were split in 20 samples, 10 from each state, with means of 5463 and 5543 cows per sample from California and New York. Mean heritability estimates for milk, fat, and protein yields for California data were, respectively, 0.34, 0.35, and 0.40 for first; 0.31, 0.33, and 0.39 for second; and 0.28, 0.31, and 0.37 for third lactations. For New York data, estimates were 0.35, 0.40, and 0.34 for first; 0.34, 0.44, and 0.38 for second; and 0.32, 0.43, and 0.38 for third lactations. Means of estimates of genetic correlations between first and second, first and third, and second and third lactations for California data were 0.86, 0.77, and 0.96 for milk; 0.89, 0.84, and 0.97 for fat; and 0.90, 0.84, and 0.97 for protein yields. Mean estimates for New York data were 0.87, 0.81, and 0.97 for milk; 0.91, 0.86, and 0.98 for fat; and 0.88, 0.82, and 0.98 for protein yields. Environmental correlations varied from 0.30 to 0.50 and were larger between second and third lactations. Phenotypic correlations were similar for both states and varied from 0.52 to 0.66 for milk, fat and protein yields. These estimates are consistent with previous estimates obtained with animal models. Yields in different lactations are not statistically the same trait but for selection programs such yields can be modelled as the same trait because of the high genetic correlations.

Effects of soy protein and resistance exercise on body composition and blood lipids in postmenopausal women

Objective: To assess the effect of soy protein and progressive resistance training on body composition and lipids in postmenopausal women.Design: In a controlled trial, 46 postmenopausal women were randomized to one of four groups: 25 g of soy protein (SP, n = 10), 25 g of soy protein plus resistance exercise (SPE, n = 14), 25 g of maltodextrine (placebo) (PL, n = 11), or placebo plus resistance exercise (PLE, n = 11). Progressive resistance training was held three times a week for 16 weeks and included 8 exercises (3 series of 8-12 repetitions). At baseline and after 16 weeks, body mass index, waist circumference (WC), body fat, muscle mass and serum lipid levels were measured. To confirm isoflavone absorption, urinary concentrations were determined. The t-test of Student and ANOVA were used in the statistical analysis.Results: Subjects were classified as overweight and showed android fat distribution: Urinary isoflavone excretion indicated compliance to soy protein treatment. After 16 weeks of intervention, both SPE and PLE groups showed a significant increase of 1.3 kg in muscle mass and reduction in WC of -1.4 and -2.1 cm, respectively (p < 0.05). Significant decreases in the mean values of total cholesterol and LDL (-29.0 and -24.0 mg/dL, p < 0.001 and p < 0.006, respectively) were observed in the users of soy protein alone (SP).Conclusions: Soy protein supplementation did not influence the indicators of body composition. However, it exerted possible favorable effects on lipid profile in postmenopausal women. The increase in muscle mass and reduction in abdominal fat were correlated with resistance training. (c) 2006 Elsevier B.V.. All rights reserved.

Reduction of erythroid progenitors in protein-energy malnutrition

Protein-energy malnutrition is a syndrome in which anaemia together with multivitamin and mineral deficiency may be present. The pathophysiological mechanisms involved have not, however, yet been completely elucidated. The aim of the present study was to evaluate the pathophysiological processes that occur in this anaemia in animals that were submitted to protein-energy malnutrition, in particular with respect to Fe concentration and the proliferative activity of haemopoietic cells. For this, histological, histochemical, cell culture and immunophenotyping techniques were used. Two-month-old male Swiss mice were submitted to protein-energy malnutrition with a low-protein diet (20g/kg) compared with control diet (400 g/kg). When the experimental group had attained a 20% loss of their original body weight, the animals from both groups received, intravenously, 20IU erythropoietin every other day for 14 d. Malnourished animals showed a decrease in red blood cells, Hb concentration and reticulocytopenia, as well as severe bone marrow and splenic atrophy. The results for serum Fe, total Fe-binding capacity, transferrin and erythropoietin in malnourished animals were no different from those of the control animals. Fe reserves in the spleen, liver and bone marrow were found to be greater in the malnourished animals. The mixed colony-forming unit assays revealed a smaller production of granulocyte-macrophage colony-forming units, erythroid burst-forming units, erythroid colony-forming units and CD45, CD117, CD119 and CD71 expression in the bone marrow and spleen cells of malnourished animals. These findings suggest that, in this protein-energy malnutrition model, anaemia is not caused by Fe deficiency or erythropoietin deficiency, but is a result of ineffective erythropoiesis.

Protein malnutrition does not impair glucose metabolism adaptations to exercise-training

Malnutrition is a common health problem in developing countries and is associated with alterations in glucose metabolism. In the present study we examine the effects of chronic aerobic exercise on some aspects of glucose metabolism in protein-deficient rats. Two groups of adult rats (90 days old) were used: Normal protein group (17%P)- kept on a normal protein diet during intra-uterine and postnatal life and Low protein group (6%P)- kept on a low protein diet during intrauterine and post natal life. After weaning (21 days old), half of the 17%P and 6%P rats were assigned to a Sedentary (Sed) or an Exercise-trained (Exerc = swimming, 1 hr/day, 5 days/week, supporting an overload of 5% of body weight) subgroup. The area under blood glucose concentration curve (Delta G) after an oral glucose load was higher in 17%P Sed rats (20%) than in other rats and lower in 6%P Exerc (11%) in relation to 6% Sed rats. The post-glucose increase in blood insulin (Delta I) was also higher in 17%P Sed (9%) than in other rats. on the other hand, the glucose disappearance rate after exogenous subcutaneous insulin administration (Kitt) was lower in 17%P Sed rats (66%) than in other rats. Glucose uptake by soleus muscle was higher in Exerc rats (30%) than in Sed rats. Soleus muscle glycogen synthesis was reduced in 6%P Sed rats (41%) compared to 17%P Sed rats but was restored in 6%P Exerc rats. Glycogen concentration was elevated in Exerc (32%) rats in comparison to Sed rats. The present results indicate that glucose-induced insulin release is reduced in rats fed low protein diet. This defect is counteracted by an increase in the sensitivity of the target tissues to insulin and glucose homeostasis is maintained. This adaptation allows protein deficient rats to preserve the ability to appropriately adapt to aerobic physical exercise training. (C) 2000 Elsevier B.V.

Energy and protein requirements for maintenance and growth of Boer crossbred kids

Meat production by goats has become an important livestock enterprise in several parts of the world. Nonetheless, energy and protein requirements of meat goats have not been defined thoroughly. The objective of this study was to determine the energy and protein requirements for maintenance and growth of 34 3/4 Boer x 1/4 Saanen crossbred, intact male kids (20.5 +/- 0.24 kg of initial BW). The baseline group was 7 randomly selected kids, averaging 21.2 +/- 0.36 kg of BW. An intermediate group consisted of 6 randomly selected kids, fed for ad libitum intake, that were slaughtered when they reached an average BW of 28.2 +/- 0.39 kg. The remaining kids (n = 21) were allocated randomly on d 0 to 3 levels of DMI (treatments were ad libitum or restricted to 70 or 40% of the ad libitum intake) within 7 slaughter groups. A slaughter group contained 1 kid from each treatment, and kids were slaughtered when the ad libitum treatment kid reached 35 kg of BW. Individual body components (head plus feet, hide, internal organs plus blood, and carcass) were weighed, ground, mixed, and subsampled for chemical analyses. Initial body composition was determined using equations developed from the composition of the baseline kids. The calculated daily maintenance requirement for NE was 77.3 +/- 1.05 kcal/kg(0.75) of empty BW (EBW) or 67.4 +/- 1.04 kcal/kg(0.75) of shrunk BW. The daily ME requirement for maintenance (118.1 kcal/g(0.75) of EBW or 103.0 kcal/kg(0.75) of shrunk BW) was calculated by iteration, assuming that the heat produced was equal to the ME intake at maintenance. The partial efficiency of use of ME for NE below maintenance was 0.65. A value of 2.44 +/- 0.4 g of net protein/kg(0.75) of EBW for daily maintenance was determined. Net energy requirements for growth ranged from 2.55 to 3.0 Mcal/kg of EBW gain at 20 and 35 kg of BW, and net protein requirements for growth ranged from 178.8 to 185.2 g/kg of EBW gain. These results suggest that NE and net protein requirements for growing meat goats exceed the requirements previously published for dairy goats. Moreover, results from this study suggest that the N requirement for maintenance for growing goats is greater than the established recommendations.

Hepatic lesions in protein-deficient adult rats

Four groups of 10 young adult Wistar male rats were fed ad libitum on a protein-free diet for periods of 7, 28, 56 and 84 days. Control groups were fed on a 20% casein diet. Food intake and body weights of rats were registered. Plasma protein levels and liver weight and fat content were determined. Sections of the caudate lobe were studied histologically. Fatty changes were classified in three grades. Protein-deficient rats exhibited loss of body weight and had low levels of plasma protein concentration. Liver lost weight after 7 days of protein deficiency; there was a gradual reduction in liver weight as periods of protein deprivation were longer. After 7 days, liver fat concentration was not significantly higher than in the respective control group; it was significantly higher in all the other malnourished animals, As periods of protein deprivation were longer, fatty changes became more severe. Other hepatic lesions were found in 5 of the 10 rats submitted to the longest period of protein deficiency. One of the rats showed a diffuse cellular atrophy, 2 animals showed an extensive haemorrhagic necrosis, another showed a focal area of reticulum collapse and the last exhibited a distortion of the normal architecture of the liver due to diffuse reticulum collapse and early nodular regeneration; these 2 last rats showed early fibrosis in portal areas. The findings suggest that other deficiencies may complicate the protein deficiency when rats are given a protein-free diet over prolonged periods. Even if the protein-deficient diet has protective nutrients, it may be that, when rats eat less food, as occurs in prolonged experiments deficiency of one or all of these elements can occur, depending on their relative amount in diet.

Morphometric study of the small intestinal mucosa in young, adult, and old rats submitted to protein deficiency and rehabilitation

Linear and stereological morphometric methods were applied to the jejunal and ileal mucosa of young, adult, and old male Wistar rats submitted to protein deficiency and rehabilitation. The animals were fed ad libitum a 2% casein diet during 42 days and then received a 20% casein diet for 30 days. Food intake, body weights, and plasma protein concentrations were recorded. In the young protein deficient rats values of mucosal height, surface area, and volume of the lamina propria were significantly lower than those of their age controls in both jejunum and ileum. In adults the differences were less marked and in the old rats all parameters were found to be unaltered by the protein deficient diet. The surface-to-volume ratio showed no significant differences between control and protein deficient in all three age groups, meaning that villus pattern did not change with protein deficiency. On rehabilitation, a striking difference between jejunum and ileum was observed in the young rats; all parameters returned to control levels in the jejunum, while they remained lower than those of their controls in the ileum.

Importance of early and continuous use of protein restriction on the progression of adriamycin nephropathy

Three experimental protocols were carried out with the aim of evaluating the role of protein restriction on the progression of the established adriamycin-induced nephropathy, and whether the protective effect of the diet persists after the diet is discontinued. The effect of a low protein diet (LPD) was studied for 6 weeks in protocol 1, 16 weeks in protocol 2 and for 28 weeks in protocol 3. In protocol 3, one group (LL) received LPD and another (NN) was given a normal protein diet (NPD). A third group (LN) received LPD for 16 weeks and then NPD for 12 weeks and a fourth group (NL) was fed NPD for 16 weeks and then LPD for 12 weeks. In protocol I the tubulo- interstitial index (TILl) of rats on LPD (Md = 2, P25 = 0.0; P75 = 3.5) after six weeks, was smaller than that of the animals on NPD (Md = 6.0; P25 = 3.0; P75 = 8.0; p < 0.05). In protocol 2, the group taking LPD presented an area of interstitial fibrosis (IF) (Md= 0.5%, P25 0.2%; P75 = 1.9%) smaller than that of the NPD group (Md = 6.8%; P25 = 5.2%; P75 = 7.1%; P < 0.05). No significant difference in the area of glomerulosclerosis (GSA) was observed between the animals on LPD (Md = 0.0%; P25 = 0.0%, P75 = 0.0%) and NPD (Md = 0.37%; P25 = 02% P75 = 1.25%; p > 0.05). In protocol 3, the group LL showed GSA (Md = 1.3%; P25 0.6%, P75 = 2.5%) and IF (Md = 3.60/0; P25 = 1.6%; P75 = 5.9%) smaller that those of LN (GSA Md = 10.1%; P25 = 6.6%; P75 = 14.8%; IF; Md = 17.3%; P25 = 14.1%; P75 = 24,5%), NL (GSA: Md = 9.1%; P25 = 5,8%; P75 = 11.7%; IF; Md = 25.0%; P25 = 20.4%; P75 = 30%), and NN (GSA: Md = 6. 75%; P25 = 4.9%; P75 = 11.7%; IF: Md = 20.9%; P25 = 16.2%; P75 = 32.4%). In conclusion, in order to be effective, LPD must be introduced early and maintained for a long period of tune.

Influence of dietary protein level on the broiler chicken's response to methionine and betaine supplements

Two experiments were conducted to compare broiler chicken responses to methionine and betaine supplements when fed diets with low protein and relatively high metabolizable energy levels (17%, 3.3 kcal/g) or moderate protein and lower metabolizable energy levels (24%, 3.0 kcal/g), resulting in different levels of carcass fat. In Experiment 1, the basal diets were formulated with corn, soybean meal, poultry by-product meal, and poultry oil. In Experiment 2, glucose monohydrate was also added, so that identical amino acid profiles could be maintained in the 17 and 24% protein diets. On average, feeding the 17 vs. 24% protein diet decreased 21-d body weight gain by 20%, increased feed conversion ratio (FCR) by 13%, and increased abdominal fat pad weight by 104%. Methionine and betaine supplements improved the performance of chicks fed the 24% protein diet in both experiments, as indicated by body weight gain and FCR. Only supplementary methionine increased performance of chicks fed 17% protein diets, and then only in Experiment 2. Neither methionine nor betaine decreased abdominal fat pad size in either experiment. Methionine supplementation decreased relative liver size and increased breast muscle protein. Both methionine and betaine increased sample feather weight, but when expressed as a percentage of body weight, no significant differences were detected. It is concluded that increasing carcass fat by manipulating percentage dietary protein level or amino acid balance does not influence betaine's activity as a lipotropic agent.

Protein requirements in larvae and adults of Scaptotrigona postica (Hymenoptera: Apidia, Meliponinae): Midgut proteolytic activity and pollen digestion

The number and degree of digestion of pollen grains in the midgut and rectum, the midgut proteolytic activity and the time of pollen grain passage through the digestive tract in the stingless bee Scaptotrigona postica (Latreille) have been analyzed. The results show similar protein requirements among larvae, nurse bees and queens, as well as between forager bees and old males, but these requirements are higher in individuals from the former groups than in those from the latter. Although protein requirements have been demonstrated to vary according to a bee's activity in the colony, they are similar among bees from different castes or sexes. These changes in feeding behavior are related to the bee's function and to less competition for nourishment among individuals of the colony. It is also noted that pollen grains took between 6 and 28 h to pass through the digestive tract. Pollen grains are irregularly accumulated in the various regions of the midgut, which may reflect functional differentiation throughout the midgut. © 2001 Elsevier B.V.

Changes in protein fractions, trypsin inhibitor and proteolytic activity in the cotyledons of germinating chickpea

The chickpea seed germination was carried out in 6 days. During the period it was observed a little variation on total nitrogen contents, however the non protein nitrogen was double. A decrease of 19.1 and 20.6% in relation to total nitrogen was observed to the total globulin and albumin fractions, respectively. The gel filtration chromatography on Sepharose CL-6B and SDS-PAGE demonstrated alterations on the distribution patterns of the albumin and total globulin fractions between the initial and the sixth day of germination suggesting the occurrence of protein degradation in the germination process.The assay for acid protease only appeared in the albumin fraction with casein and chickpea total globulin as substrates, whereas the former was more degradated than the latter, however the transformations detected in the protein fractions apppear indicated that others enzymes could be acting during the process. The trypsin inhibitor activity had a little drop after six day of germination indicating a possible increase on the digestibility of the proteins.

Protein Deficiency Attenuates the Effects of Alloxan on Insulin Secretion and Glucose Homeostasis in Rats

We have investigated the effect of alloxan on insulin secretion and glucose homeostasis in rats maintained on a 17% protein (normal protein, NP) or 6% protein (low protein, LP) diet from weaning (21 days old) to adulthood (90 days old). The incidence of alloxan diabetes was higher in the NP (3.5 times) than in the LP group. During an oral glucose tolerance test, the area under serum glucose curve was lower in LP (57%) than in NP rats while there were no differences between the two groups in the area under serum insulin curve. The serum glucose disappearance rate (Kitt) after exogenous insulin administration was higher in LP (50%) than in NP rats. In pancreatic islets isolated from rats not injected with alloxan, acute exposure to alloxan (0.05 mmol/L) reduced the glucose- or arginine-stimulated insulin secretion of NP islets by 78% and 56%, respectively, whereas for islets from LP rats, the reduction was 47% and 17% in the presence of glucose and arginine, respectively. Alloxan treatment reduced the glucose oxidation in islets from LP rats to a lesser extent than in NP islets (23% vs. 56%). In conclusion, alloxan was less effective in producing hyperglycemia in rats fed a low protein diet than in normal diet rats. This effect is attributable to an increased peripheral sensivity to insulin in addition to a better preservation of glucose oxidation and insulin secretion in islets from rats fed a low protein diet.

Genetic interactions of yeast eukaryotic translation initiation factor 5a (eIF5A) reveal connections to poly(A)-binding protein and protein kinase C signaling

The highly conserved eukaryotic translation initiation factor eIF5A has been proposed to have various roles in the cell, from translation to mRNA decay to nuclear protein export. To further our understanding of this essential protein, three temperature-sensitive alleles of the yeast TIF51A gene have been characterized. Two mutant eIF5A proteins contain mutations in a proline residue at the junction between the two eIFSA domains and the third, strongest allele encodes a protein with a single mutation in each domain, both of which are required for the growth defect. The stronger tif51A alleles cause defects in degradation of short-lived mRNAs, supporting a role for this protein in mRNA decay. A multicopy suppressor screen revealed six genes, the overexpression of which allows growth of a tif51A-1 strain at high temperature; these genes include PAB1, PKC1, and PKC1 regulators WSC1, WSC2, and WSC3. Further results suggest that eIFSA may also be involved in ribosomal synthesis and the WSC/PKC1 signaling pathway for cell wall integrity or related processes.