Comparison of lymphatic and venous interpositional autografts in experimental microsurgery of the canine lymphatics

In mongrel dogs, 56 autologous lymphatic and vein grafts were interpositioned to bridge a defect in the femoral collecting lymphatics. In one group, 26 lymphatic autografts were interpositioned with good results. No obstruction was observed over 6 months. In another group, 20 venous autografts were interpositioned after irrigation with heparinized saline and another 10 autografts were interpositioned without irrigation. After 1 week, four irrigated grafts were partially occluded with a red thrombus; after 6 months, all grafts were totally occluded. In a third group, 15 lymphaticolymphatic anastomoses were enveloped by a silicone sheet to provoke prolonged devascularization. None of the vessels was patent. Anastomotic patency was inspected in vivo postoperatively. The specimens were studied with light microscopy and scanning and transmission electron microscopy. Prolonged devascularization damaged the endothelial cells. The results show that the lymphatic vessel autograft is the best choice for an interpositional autografting to bridge a defect in lymphatic vessels.     

The Presence of Actinidin (Cysteine Protease) and Recombinant Plasmids Carrying the Actinidin Gene Influence the Growth of Saccharomyces Cerevisiae

Actinidin is a protease found abundantly in the fruit of Actinidia chinensis or Kiwi fruit. The overproduction of this protein in microorganisms, especially using the yeast Saccharomyces cerevisiae, would be economically valuable as it would simplify the extraction and purification of the protein. It was observed, however, that the yeast growth rate was reduced by the presence of externally supplied actinidin in the growth medium. It was also observed that actinidin present in the yeast growth medium was partially degraded. Several actinidin-encoding gene variants have been cloned in a yeast expression and secretion vector. It was observed that different actinidin gene constructions influenced the growth rate of S. cerevisiae in complete media. Recombinant plasmids carrying only the mature actinidin-encoding DNA sequences reduced yeast transformability significantly and had the least stability. The results thus suggest that the presence of a recombinant plasmid carrying a gene of a potentially toxic protein may result in a deleterious effect on the host cell.     

Schistosomiasis in Indonesia: past and present

Schistosomiasis is endemic in Indonesia in two isolated areas, Lindu valley and Napu valley, both located in the Province of Central Sulawesi. In 1940, a prevalence survey was initiated in Lake Lindu, which indicated a Schistosoma japonicum infection prevalence of 56% among the population of Anca, Tomado and Langko villages. Another survey was conducted in 1973 in Napu valley and very high infection prevalences of up to 72% were found among the population in Winowanga village. Since then, comprehensive studies on the epidemiology and the effects of control have been carried out in 24 endemic villages in both areas. Over the past six decades, schistosomiasis control has been implemented and the average prevalence is now much lower than before the control programme was launched. In 2006, it was 0.49% in 7 villages in Lindu valley. In Napu valley, the average infection prevalence among the population of 17 villages was 1.08% in the same year. Again in 2006, the prevalence of infection in snails ranged from 0 to 13.4% and from 0 to 9.1% in Napu and Lindu valleys, respectively. The highest prevalence among snails was found in Dodolo village. The prevalence of S. japonicum in the reservoir host Rattus spp. ranged from 0 to 20% and the highest prevalence was again found in Dodolo village. Contemporary data suggest that transmission of schistosomiasis is still ongoing in Indonesia despite regular surveillance and control activities covering the whole endemic area.     

The Role of Amino-Terminal and Carboxy-Terminal Extensions in the Processing and Translocation of a Plant Proteinase (Actinidin) Expressed in the Yeast Saccharomyces cerevisiae

Summary A plant proteinase gene naturally occuring in the Kiwi fruit plant (Actinidia chinensis) has been expressed in a yeast Saccharomyces cerevisiae. Different gene constructions consisting of different portions of the whole actinidin-encoding gene have been created and expressed using an expression-secretion yeast vector. It was observed that the amino- and carboxy-terminal extensions of the actinidin-encoding gene were required for the correct expression of the gene in yeast. A gene construction lacking both amino- and C-terminal extensions did not result in a detectable protein product. Similarly, a gene construction consisting of the amino-terminal extension plus mature actinidin-encoding DNA did not result in a detectable expression. However, intracellular expression was observed when a gene construction consisting of mature actinidin-encoding DNA plus C-terminal extension portion was employed. The expressed polypeptide was found however not to be correctly processed as it had a bigger size than the native actinidin. The correctly processed polypeptide was expressed intracellularly when the full-length actinidin cDNA was expressed in a vacuolar protease-proficient yeast strain. However, when a vacuolar protease-deficient yeast strain was employed, it was found that the precursor protein was not correctly processed, suggesting that the actinidin precursor had entered the vacuole and undergone proteolytic processing. The full-length actinidin cDNA consisted of the amino-terminal extension DNA, mature actinidin-encoding DNA, and C-terminal extension DNA. The results thus suggested that both amino- and C-terminal extensions were required for correct expression and processing of actinidin in yeast. The intracellular expression also suggested that the actinidin-encoding sequences contain intracellular targeting sequences which override the secretion signal included in the expression-secretion vector.     

Metabolism of betaine as a carbon source by an osmotolerant bacterium isolated from the weed rhizosphere

An isolate of an osmotolerant rhizobacterium has been obtained from a weed rhizosphere which showed tolerance up to 1.0 M NaCl. The isolate has been subjected to growth analysis in a medium which contained 10 mM betaine as the sole carbon source. It was observed that betaine could be used as the sole carbon source for the growth of salt-tolerant rhizobacteria under NaCl-stress at 1.0 M concentration. Interestingly, it was found that betaine at 100 mM concentration suppressed the growth of salt-tolerant rhizobacteria. The growth of the osmotolerant rhizobacterium was stimulated when it was grown in a medium containing both glucose and betaine, demonstrating that betaine was an osmoprotectant. The presence of glucose at 10 mM concentration, however, did not alleviate the growth-suppressive effect of betaine at 100 mM concentration. The osmoprotective effect of betaine was demonstrated by the fact that the addition of betaine at different time intervals enhanced the growth accordingly. However, the growth-suppressive effect of betaine at 100 mM concentration was also noticeable when betaine was added at different time intervals.     

Unsupervised machine-learning method for improving the performance of ambulatory fall-detection systems

Background  

Falls can cause trauma, disability and death among older people. Ambulatory accelerometer devices are currently capable of detecting falls in a controlled environment. However, research suggests that most current approaches can tend to have insufficient sensitivity and specificity in non-laboratory environments, in part because impacts can be experienced as part of ordinary daily living activities.     

Role of dendritic cells in antibody-dependent enhancement of dengue virus infection

Dengue viruses (DV), composed of four distinct serotypes (DV1 to DV4), cause 50 to 100 million infections annually. Durable homotypic immunity follows infection but may predispose to severe subsequent heterotypic infections, a risk conferred in part by the immune response itself. Antibody-dependent enhancement (ADE), a process best described in vitro, is epidemiologically linked to complicated DV infections, especially in Southeast Asia. Here we report for the first time the ADE phenomenon in primary human dendritic cells (DC), early targets of DV infection, and human cell lines bearing Fc receptors. We show that ADE is inversely correlated with surface expression of DC-SIGN (DC-specific intercellular adhesion molecule-3-grabbing nonintegrin) and requires Fc gamma receptor IIa (FcgammaRIIa). Mature DC exhibited ADE, whereas immature DC, expressing higher levels of DC-SIGN and similar FcgammaRIIa levels, did not undergo ADE. ADE results in increased intracellular de novo DV protein synthesis, increased viral RNA production and release, and increased infectivity of the supernatants in mature DC. Interestingly, tumor necrosis factor alpha and interleukin-6 (IL-6), but not IL-10 and gamma interferon, were released in the presence of dengue patient sera but generally only at enhancement titers, suggesting a signaling component of ADE. FcgammaRIIa inhibition with monoclonal antibodies abrogated ADE and associated downstream consequences. DV versatility in entry routes (FcgammaRIIa or DC-SIGN) in mature DC broadens target options and suggests additional ways for DC to contribute to the pathogenesis of severe DV infection. Studying the cellular targets of DV infection and their susceptibility to ADE will aid our understanding of complex disease and contribute to the field of vaccine development.     

Study of the effects of temperature and pH on lactic acid production from fresh cassava roots in tofu liquid waste by Streptococcus bovis

We report a study of the effects of temperature and pH on the kinetics of lactic acid production from fresh cassava roots (FCR) by Streptococcus bovis in new media: tofu liquid waste (TLW); TLW with 2 wt% concentrated maguro waste (TLW + CMW2); and in a standard trypto-soya broth (TSB) compared with a standard medium (glucose in TSB). The results showed that 39 °C and pH 5.5 were optimal for fermentation properties (lactic acid concentration, productivity and specific growth rate) in all media, including the standard. The energy of activation (E_a) and the energy of deactivation (E_d) of lactic acid fermentation in all media were calculated using the Arrhenius relation. The E_a and E_d values increased in the order FCR in TLW < FCR in TLW + CMW2 < glucose in TSB < FCR in TSB, while values of productivity and specific growth rate decreased in the same order. The effects of pH on productivity and specific growth rate were evaluated by Michaelis–Menten and Monod-type equations. For pH, the inhibition is competitive for both productivity and specific growth rate. By controlling the pH at 5.80, the inhibition for both productivity and specific growth rate were minimized in all media.