Biosynthesis of lysosomal hydrolases: their synthesis in bound polysomes and the role of co- and post-translational processing in determining their subcellular distribution

By in vitro translation of mRNA’s isolated from free and membrane-bound polysomes, direct evidence was obtained for the synthesis of two lysosomal hydrolases, β-glucuronidase of the rat preputial gland and cathespin D of mouse spleen, on polysomes bound to rough endoplasmic reticulum (ER) membranes. When the mRNA’s for these two proteins were translated in the presence of microsomal membranes, the in vitro synthesized polypeptides were cotranslationally glycosylated and transferred into the microsomal lumen. Polypeptides synthesized in the absence of microsomal membranes were approximately 2,000 daltons larger than the respective unglycosylated microsomal polypeptides found after short times of labeling in cultured rat liver cells treated with tunicamycin. This strongly suggests that nascent chains of the lysosomal enzymes bear transient amino terminal signals which determine synthesis on bound polysomes and are removed during the cotranslational insertion of the polypeptides into the ER membranes. In the line of cultured rat liver cells used for this work, newly synthesized lysosomal hydrolases showed a dual destination; approximately 60 percent of the microsomal polypeptides detected after short times of labeling were subsequently processed proteolytically to lower molecular weight forms characteristic of the mature enzymes. The remainder was secreted from the cells without further proteolytic processing. As previously observed by other investigations in cultured fibroblasts (A. Gonzalez-Noriega, J.H. Grubbs, V. Talkad, and W.S. Sly, 1980, J Cell Biol. 85: 839-852; A. Hasilik and E.F. Neufeld, 1980, J. Biol. Chem., 255:4937-4945.) the lysosomotropic amine chloroquine prevented the proteolytic maturation of newly synthesized hydrolases and enhanced their section. In addition, unglycosylated hydrolases synthesized in cells treated with tunicamycin were exclusively exported from the cells without undergoing proteolytic processing. These results support the notions that modified sugar residues serve as sorting out signals which address the hydrolases to their lysosomal destination and that final proteolytic cleavage of hydrolase precursors take place within lysosome itself. Structural differences in the carbohydrate chains of intracellular and secreted precursors of cathespin D were detected from their differential sensitivity to digestion with endoglycosidases H and D. These observations suggest that the hydrolases exported into the medium follow the normal secretory route and that some of their oligosaccharides are subject to modifications known to affect many secretory glycoproteins during their passage through the Golgi apparatus.     

Noisy-threshold control of cell death

Background  

Cellular responses to death-promoting stimuli typically proceed through a differentiated multistage process, involving a lag phase, extensive death, and potential adaptation. Deregulation of this chain of events is at the root of many diseases. Improper adaptation is particularly important because it allows cell sub-populations to survive even in the continuous presence of death conditions, which results, among others, in the eventual failure of many targeted anticancer therapies.     

Evolution of dentition in prehistoric Ohio Valley Native Americans III. Metrics of deciduous dentition

Deciduous tooth size in Native Americans of the Ohio Valley area is fairly stable from the terminal Late Archaic (3200 BP) through the Late Prehistoric (350 BP) periods. Some fluctuation in average size did occur during this time. These fluctuations most likely reflect random changes due to gene drift. However, no difference in the pattern of interactions among the sizes of teeth (covariance structures) can be demonstrated during this period. Principal components analysis of the buccolingual and mesiodistal dimensions in the total sample indicate that the major axis of deciduous tooth size in the Ohio Valley population shows an allometric relationship, with the dimensions of the anterior teeth increasing (or decreasing) as the 1.33 power of the dimension of m1(1) and as the 2.0 power of the dimension of m2(2). Comparison of the Ohio Valley samples with other samples from the Eastern Woodlands suggests that geography may have played a minor role in structuring deciduous tooth size variation. For the most part, however, widely separated Eastern Woodlands populations appear to have been evolving independently with respect to deciduous tooth size.     

Evidence for long‐term migration on the Balkan Peninsula using dental and cranial nonmetric data: Early interaction between Corinth (Greece) and its colony at Apollonia (Albania)

This article seeks to identify “Greeks” and “non‐Greeks” in “mixed” mortuary contexts in a Greek colony. Specifically, we test the hypothesis that Illyrian and Greek individuals lived and were buried together at the Corinthian colony of Apollonia, Albania (established ca. 600 BC). The pattern of human biological interaction at Apollonia is tested by identifying variation in genetic relatedness using biodistance analysis of dental and cranial nonmetric traits for three sites: Apollonia (n = 116), its founder‐city Corinth (n = 69), and Lofkënd (n = 108), an inland site near Apollonia pre‐dating colonization. Logistic regression analysis estimates that individuals from colonial Apollonia are most closely related to prehistoric Illyrian populations (from Lofkënd and prehistoric Apollonia), rather than Greeks (from Corinth). The phenotypic similarity between colonial Apollonia and prehistoric Illyria suggests that there was a large Illyrian contribution to the gene pool at the colony of Apollonia. However, some trait combinations show low biological distances among all groups, suggesting homogeneity among Illyrian and Greek populations (assessed through pseudo‐Mahalanobis' D²). The degree of phenotypic similarity suggests shared ancestry and long‐term migration throughout these regions. The impacts of missing data and small sample sizes are also considered. Am J Phys Anthropol 153:236–248, 2014. © 2013 Wiley Periodicals, Inc.     

Estimating body mass in subadult human skeletons

Methods for estimating body mass from the human skeleton are often required for research in biological or forensic anthropology. There are currently only two methods for estimating body mass in subadults: the width of the distal femur metaphysis is useful for individuals 1–12 years of age and the femoral head is useful for older subadults. This article provides age‐structured formulas for estimating subadult body mass using midshaft femur cross‐sectional geometry (polar second moments of area). The formulas were developed using data from the Denver Growth Study and their accuracy was examined using an independent sample from Franklin County, Ohio. Body mass estimates from the midshaft were compared with estimates from the width of the distal metaphysis of the femur. Results indicate that accuracy and bias of estimates from the midshaft and the distal end of the femur are similar for this contemporary cadaver sample. While clinical research has demonstrated that body mass is one principle factor shaping cross‐sectional geometry of the subadult midshaft femur, clearly other biomechanical forces, such as activity level, also play a role. Thus formulas for estimating body mass from femoral measurements should be tested on subadult populations from diverse ecological and cultural circumstances to better understand the relationship between body mass, activity, diet, and morphology during ontogeny. Am J Phys Anthropol 143:146–150, 2010. © 2010 Wiley‐Liss, Inc.     

Evaluation of juvenile stature and body mass prediction

This investigation evaluates the performance of juvenile stature (from tibia and radius lengths) and body mass (from breadth of the femoral distal metaphysis) prediction equations based on the Denver Growth Study sample (Ruff C. 2007. Am J Phys Anthropol 133 698-716). The sample used here for evaluation is an independent sample of juveniles brought to the Franklin County (Ohio) Coroner in 1990-1991. The Ohio sample differs somewhat from the Denver reference sample: it includes approximately 25% African-Americans (rather than all European-Americans), a significant number of right limb bones were measured (rather than all left side), it includes a wider range of economic statuses and it includes individuals who died from disease and trauma. As such the composition and measures of the Ohio sample correspond more generally to that seen in skeletal samples so that the accuracy of the estimates from the present sample should approach those found in practical applications of these methods. Results indicate that both juvenile body mass and stature are estimated relatively accurately. Accuracy of body mass estimates for 1-13-year-old juveniles is similar for African-American and European-American males and females. The least accurate estimates are for individuals in the 8-13 years age class (excluding individuals with body mass indices greater than the age specific 95th percentile): n = 9, +/- 2.9 kg, 95% confidence interval 1.4-4.4 kg. Accuracy of stature estimates for 1-17-year-old juveniles is comparable for the tibia and radius and, as with body mass estimates, are similar for African-American and European-American males and females. For combined age, sex, and ancestry groups average accuracies are in the +/-3.5 to +/-6.5 cm range. Some limitations of the methods are discussed.     

Monoclonal antibody to dengue capsid protein: Its application in dengue studies

Dengue fever (DF) and dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS) are considered the most important arthropod-borne viral diseases in terms of morbidity and mortality. The emergency and severity of dengue (Den) infections increase the necessity of an early, quick and effective dengue laboratory diagnostic. Viral isolation is considered a gold standard for diagnosis of dengue infection using monoclonal antibodies (mAbs) as a tool for determining serotype specificity. Alternatives have been used to improve sensitivity and time to dengue diagnosis. Based on the early expression of dengue C protein in the life cycle, we focused our study on the application of an anti-dengue 2 virus capsid protein mAb in dengue diagnosis. The kinetic expression of dengue-2 capsid in mosquito cells and its immuno-localization in experimentally infected suckling albin Swiss (OF-1) mice brain tissues was established. The results demonstrate the possible utility of this mAb in early dengue diagnosis versus traditional isolation. In addition, a preliminary study of an enzyme immunoassay method using 8H8 mAb for specific detection of dengue C protein antigen was performed, making possible recombinant C protein quantification. The results suggest that detection of dengue capsid protein could be useful in the diagnosis of early dengue infection.Key words: monoclonal antibodies, capsid protein, dengue virus, diagnosis, immunoassays     

Evaluation of a vermicompost and leachates on <i>Solidago x hybrida</i> and organic carbon mineralization under aerobic incubations

In the floriculture region of Tenancingo in the State of Mexico, the application of stabilized organic matter, such as vermicompost and leachates, contributes to improve the quality of the soil and plant nutrition. However, it is important to know the chemical composition of a vermicompost and the mineralization process. This is because the amount and speed of nutrient release which will be available to the crop will depend on that knowledge. The objective of the study was to evaluate the effect of the application of a vermicompost and leachates on various quantitative variables of Solidago x hybrid, and the mineralization of organic carbon under aerobic incubations. Vermicompost (74 and 36 g/kg soil), leachates (5 and 10 L/kg soil), 0.33 g/kg soil of chemical fertilizer Ca (NO₃)₂, and not treated soil (control) were applied under greenhouse conditions to evaluate their effects on plant growth variables. Mixtures of 100 g of soil with vermicompost and leachates were made in the laboratory which were incubated during 9 weeks to obtain the potentially mineralizable organic carbon (Corg PM) and the rate of mineralization (k) after adjusting an exponential model. In the greenhouse experiment there were no statistical differences after applying vermicompost and leachates on the quantitative variables (number of stems per plant, diameter of the panicle, fresh weight, plant length and stem diameter) with respect to the control (p>0.05). The effect among the applied doses was evident only for variables such as fresh weight, panicle length and stem diameter with respect to the control. In incubated soils, k values ranged between 0.209-0.325 C mg/kg soil/week. Only with the application of leachates in high doses two pools of organic matter were shown: one soluble labile (102.9 mg/kg soil) and the other hydrolysable (819 mg/kg soil). The soluble, labile fraction favored nutrient availability immediately after its application to the soil. However, a single pool of hydrolysable organic C (987-1074 mg/kg soil) was found when vermicompost was applied. It was associated with a release of organic matter during crop development, and to a possible stimulation of microbial activity. High values of electrical conductivity in vermicompost and leachates (8.2-11.7 mS/m) suggest a moderate application of both products.     

蚕豆叶片发育与衰老过程中超氧物歧化酶活性与丙二醛含量变化

蚕豆植株叶片随茎节自上而下表现出明显的发育与衰老顺序,可作为衰老特征的是叶绿素和蛋白质含量明显下降。蚕豆叶中SOD活性主要定位于12 000× g离心后所得的上清液和叶绿体组分。衰老叶片的SOD总活性和叶绿体组分的相对活性都有所下降,SOD同工酶谱也发生了改变。O_2~ 产生速率随叶龄增大而稍上升;而MDA含量在叶片外观表现枯黄衰老征兆前就急剧上升。可能因为衰老叶片过氧化氢酶活性大幅度下降与SOD之间的不平衡,致使O_2~ 代谢中间产物累积而引起膜的损伤.