Transient active force generation and stress fibre remodelling in cells under cyclic loading

Biomechanics and Modeling in Mechanobiology - The active cytoskeleton is known to play an important mechanistic role in cellular structure, spreading, and contractility. Contractility is actively...     

Management of psychiatric problems in a Kenyan mission hospital.

Many psychiatric conditions are found in the rural general hospital in Kenya and are recognisable and manageable along standard Western lines. An essential element in management is the training of the nursing staff. The psychiatric patient should be actively managed and can benefit from the unsophisticated treatment possible at the mission hospital.     

Aberrant fetal growth and development after in vitro culture of sheep zygotes.

The effects of in vitro culture systems for sheep zygotes on subsequent fetal growth and development to day 61 and day 125 of gestation were studied. Zygotes recovered from superovulated Scottish Blackface ewes approximately 36 h after intrauterine insemination using semen from a single Suffolk sire were cultured for 5 days in (a) a granulosa cell co-culture system (co-culture); (b) synthetic oviductal fluid medium without serum (SOF-); and (c) synthetic oviductal fluid medium supplemented with human serum (SOF+). Control embryos were recovered from superovulated donor ewes at day 6 after oestrus. Embryos were transferred at day 6 to synchronous Scottish Blackface recipient ewes. In total, 146 gravid uteri were recovered, comprising 97 at day 61 (20 co-culture, 27 SOF-, 25 SOF+ and 25 control) and 49 at day 125 (13 co-culture, 8 SOF-, 6 SOF+ and 22 control) of gestation. Fetuses derived from co-cultured embryos were 14% heavier (P < 0.01) by day 61 of gestation than those derived from control embryos. Growth coefficients derived from the linear allometric equation logey = logea + b logex (where y = organ mass; x = fetal mass) were significantly greater (P < 0.05) for liver, heart, kidneys and plantaris muscle in fetuses derived from co-cultured embryos, and for liver in fetuses derived from SOF+ embryos than those for control fetuses. Fetuses derived from co-cultured embryos were 34% heavier (P < 0.001) and fetuses derived from SOF+ embryos were 18% heavier (P < 0.01) by day 125 of gestation than those derived from control embryos. Growth coefficients for liver and heart for fetuses derived from co-culture and SOF+ embryos were also significantly greater (P < 0.05) at this stage of gestation than those for control group fetuses. In contrast, allometric coefficients for these organs in fetuses derived from embryos cultured in SOF without serum supplementation were not different from those for controls. Excessive volumes of amniotic fluid (polyhydramnios) were observed in 23% of conceptuses derived from co-cultured embryos. In vitro embryo culture can significantly influence fetal growth and this study provides quantitative evidence of major shifts in the patterns of organ and tissue development.     

Examining the Role of Testosterone in Mediating Short-Term Aggressive Responses to Social Stimuli in a Lizard

Hormones have been suggested as a key proximate mechanism that organize and maintain consistent individual differences in behavioural traits such as aggression. The steroid hormone testosterone in particular has an important activational role in mediating short-term aggressive responses to social and environmental stimuli within many vertebrate systems. We conducted two complementary experiments designed to investigate the activational relationship between testosterone and aggression in male Egernia whitii, a social lizard species. First, we investigated whether a conspecific aggressive challenge induced a testosterone response and second, we artificially manipulated testosterone concentrations to examine whether this changed aggression levels. We found that at the mean level, plasma T concentration did not appear to be influenced by an aggression challenge. However, there was a slight indication that receiving a challenge may influence intra-individual consistency of plasma T concentrations, with individuals not receiving an aggression challenge maintaining consistency in their circulating testosterone concentrations, while those individuals that received a challenge did not. Manipulating circulating testosterone concentrations had no influence on either mean-level or individual-level aggression. Combined with our previous work, our study adds increasing evidence that the relationship between testosterone and aggression is not straightforward, and promotes the investigation of alternative hormonal pathways and differences in neuro-synthesis and neuroendocrine pathways to account for species variable testosterone - aggression links.     

A novel member of the RNase D exoribonuclease family functions in mitochondrial guide RNA metabolism in Trypanosoma brucei

RNA turnover and RNA editing are essential for regulation of mitochondrial gene expression in Trypanosoma brucei. RNA turnover is controlled in part by RNA 3' adenylation and uridylation status, with trans-acting factors also impacting RNA homeostasis. However, little is known about the mitochondrial degradation machinery or its regulation in T. brucei. We have identified a mitochondrial exoribonuclease, TbRND, whose expression is highly up-regulated in the insect proliferative stage of the parasite. TbRND shares sequence similarity with RNase D family enzymes but differs from all reported members of this family in possessing a CCHC zinc finger domain. In vitro, TbRND exhibits 3' to 5' exoribonuclease activity, with specificity toward uridine homopolymers, including the 3' oligo(U) tails of guide RNAs (gRNAs) that provide the sequence information for RNA editing. Several lines of evidence generated from RNAi-mediated knockdown and overexpression cell lines indicate that TbRND functions in gRNA metabolism in vivo. First, TbRND depletion results in gRNA tails extended by 2-3 nucleotides on average. Second, overexpression of wild type but not catalytically inactive TbRND results in a substantial decrease in the total gRNA population and a consequent inhibition of RNA editing. The observed effects on the gRNA population are specific as rRNAs, which are also 3'-uridylated, are unaffected by TbRND depletion or overexpression. Finally, we show that gRNA binding proteins co-purify with TbRND. In summary, TbRND is a novel 3' to 5' exoribonuclease that appears to have evolved a function highly specific to the mitochondrion of trypanosomes.     

Regulation of the inner membrane mitochondrial permeability transition by the outer membrane translocator protein (peripheral benzodiazepine receptor)

We studied the properties of the permeability transition pore (PTP) in rat liver mitochondria and in mitoplasts retaining inner membrane ultrastructure and energy-linked functions. Like mitochondria, mitoplasts readily underwent a permeability transition following Ca(2+) uptake in a process that maintained sensitivity to cyclosporin A. On the other hand, major differences between mitochondria and mitoplasts emerged in PTP regulation by ligands of the outer membrane translocator protein of 18 kDa, TSPO, formerly known as the peripheral benzodiazepine receptor. Indeed, (i) in mitoplasts, the PTP could not be activated by photo-oxidation after treatment with dicarboxylic porphyrins endowed with protoporphyrin IX configuration, which bind TSPO in intact mitochondria; and (ii) mitoplasts became resistant to the PTP-inducing effects of N,N-dihexyl-2-(4-fluorophenyl)indole-3-acetamide and of other selective ligands of TSPO. Thus, the permeability transition is an inner membrane event that is regulated by the outer membrane through specific interactions with TSPO.     

Expression of Cryptosporidium parvum Cpa135/CpCCP1 chimeras in Giardia duodenalis: organization of the protein domains affects the protein secretion pathway

Cpa135 is a multidomain antigenic protein secreted at the sporozoite stage of the Apicomplexa protozoan Cryptosporidium parvum. Previous studies have shown that the protozoan flagellate parasite Giardia duodenalis is a suitable system for the heterologous expression of secreted proteins of Apicomplexa. Here, we designed three different Cpa135 variants fused to a C-terminal HA tag in order to test their expression in G. duodenalis under the control of the inducible promoter of the cyst wall protein 1 gene (cwp1). The three Cpa135 chimeras encompassed different portions of the protein; CpaG encodes the entire polypeptide of 1574 amino acids (aa); CpaGΔC includes the first 826 aa at the N-terminus; and CpaGΔN consists in of the final 833 aa at the C-terminus. Immunoblot experiments showed that CpaG and CpaGΔN maintained the epitopes recognized by anti-C. parvum-specific human serum. The intracellular localization and transport of the three Cpa135 variants were studied by immunofluorescence in combination with G. duodenalis-specific antibodies. CpaGΔC was mainly accumulated in the endoplasmic reticulum and the intact form was also excreted in the medium. Differently, the Cpa135 chimeras possessing an intact C-terminus (CpaG and CpaGΔN) were transported towards the forming cyst wall possibly and were not detected in the medium. Furthermore, the full-length CpaG was incorporated into the cyst wall. The data presented suggest that the C-terminus of Cpa135, which includes a cysteine reach domain, could influence the secretion of the chimeric proteins.     

Restricting dairy cow access time to pasture in early lactation: the effects on milk production, grazing behaviour and dry matter intake

One of the main aims of pasture-based systems of dairy production is to increase the proportion of grazed grass in the diet. This is most easily achieved by increasing the number of grazing days. However, periods of inclement weather conditions can reduce the number of days at pasture. The two objectives of this experiment were: (i) to investigate the effect of restricting pasture access time on animal production, grazing behaviour and dry matter intake (DMI) of spring calving dairy cows in early lactation; and (ii) to establish whether silage supplementation is required when cows return indoors after short grazing periods. In all, 52 Holstein-Friesian spring calving dairy cows were assigned to a four-treatment study from 25 February to 26 March 2008. The four treatments were: full-time access to pasture (22H; control); 4.5-h- pasture access after both milkings (2 × 4.5H); 3-h pasture access after both milkings (2 × 3H); 3-h pasture access after both milkings with silage supplementation by night (2 × 3SH). All treatments were offered 14.4 kg DM/cow per day herbage from swards, with a mean pre-grazing yield of 1739 kg DM/ha above 4 cm, - and were supplemented with 3 kg DM/cow per day of concentrate. The 2 × 3SH treatment was offered an additional 4 kg DM/cow of grass silage by night. Restricting pasture access time (2 × 3H, 2 × 3SH and 2 × 4.5H) had no effect on milk (28.3 kg/cow per day) and solids-corrected milk (27.2 kg/cow per day) yield when compared with the treatment grazing full time. Supplementing animals with grass silage did not increase milk production when compared with all other treatments. Milk protein concentration tended to be lower (P = 0.08; 32.2 g/kg) for the 2 × 3SH animals when compared with the 22H animals (33.7 g/kg). The grass DMI of the 2 × 3SH treatment was significantly lower (-2.3 kg DM/cow per day) than all other treatments (11.9 kg DM/cow per day), yet the total DMI of these animals was highest (16.6 kg DM/cow per day). The 22H cows grazed for 481 min/cow per day, which is significantly longer than all other treatments. The 2 × 3H animals grazed for 98% of the time, whereas the 2 × 3SH grazed for 79% of their time at pasture. Restricting pasture access time did not affect end body weight or body condition score. The results of this study indicate that restricting pasture access time of dairy cows in early lactation does not affect milk production performance. Furthermore, supplementing cows with grass silage does not increase milk production but reduces grazing efficiency.