Molecular cloning of Odf3 encoding a novel coiled-coil protein of sperm tail outer dense fibers.

The outer dense fibers (ODF) are the main cytoskeletal structures of the sperm tail found in animals with internal fecundation. They consist of at least 14 polypeptides from which only a few are identified due to difficulties in isolation of the protein components. Here we report the isolation and molecular characterization of Odf3, encoding a novel protein of rat sperm ODF. Odf3 is transcribed in testes and more specifically in spermatids but it is also expressed in epididymides and brain suggesting a possible involvement in building of the cellular cytoskeleton. Odf3 encodes a putative protein of approximately 110 kDa. Secondary structure predictions indicated that ODF3 is a coiled-coil protein. The identification of coiled-coil proteins as constituents of outer dense fibers reveals a model for ODF formation.     

Mpe1 senses the binding of pre-mRNA and controls 3′ end processing by CPF

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Statistical Analysis of the Comet Assay Using a Mixture of Gamma Distributions

Tail moments in the single cell gel electrophoresis (comet) assay usually do not follow a normal distribution, making the statistical analysis complicated. Researchers have used a wide variety of statistical techniques in an attempt to overcome this problem. In many cases, the tail moments follow a bimodal distribution that can be modeled with a mixture of gamma distributions. This bimodality may be due to cells being in two different stages of the cell cycle at the time of treatment. Maximum likelihood, modified to accommodate censored data, can be used to estimate the five parameters of the gamma mixture distribution for each slide. A weighted analysis of variance on the parameter estimates for the gamma mixtures can be performed to determine differences in DNA damage between treatments. These methods were applied to an experiment on the effect of thymidine kinase in DNA damage and repair. Analysis based on the mixture of gamma distributions was found to be more statistically valid, more powerful, and more informative than analysis based on log-transformed tail moments.     

High Thyrotropin Levels and Risk of Mortality in the Elderly With Subclinical Hypothyroidism: A Systematic Review and Meta-analysis

ObjectiveThis study aims to determine whether elevated endogenous thyrotropin levels contribute to an increased risk of adverse outcomes, such as all-cause mortality in older adults with subclinical hypothyroidism.MethodsEight electronic databases were searched for relevant articles from inception until March 23, 2022. Cohort studies assessing the association between thyrotropin levels and the risk of mortality among older adults aged ≥60 years with subclinical hypothyroidism were eligible. The outcomes of interest were either all-cause or cardiovascular-related mortality. Two independent researchers assessed the eligibility of the studies and collected data through a previously defined data extraction form. The Newcastle-Ottawa Scale was used to evaluate the quality of evidence, and multivariate-adjusted hazard ratios (HRs) (95% Cl) were collected as the necessary risk estimate for synthesis. Random-effects models were applied for meta-analysis.ResultsOverall, 13 studies involving 44 514 participants were included in this meta-analysis. There were no significant differences in the risk of all-cause mortality (pooled HR: 1.18 [95% Cl: 0.95, 1.45], I² = 94%) and cardiovascular-related mortality (pooled HR: 1.08 [95% Cl: 0.94, 1.23], I² = 0%) between euthyroid older adults and older adults with subclinical hypothyroidism. The results remained the same when only older adults with thyrotropin ≥10 mIU/L were assessed (pooled HR for all-cause mortality and cardiovascular-related mortality, respectively: 1.53 [95% Cl: 0.81, 2.88], I² = 22%, 1.35 [95% Cl: 0.63, 2.86], I² = 43%).ConclusionHigh thyrotropin levels are not associated with increased risk for all-cause mortality as well as cardiovascular-related mortality in older adults aged ≥60 years with subclinical hypothyroidism, suggesting an unnecessity in initialing treatment.     

Prevalence of an inflammation and necrosis syndrome in suckling piglets

The current study describes the results obtained from clinical examination of over 4700 suckling piglets from 19 individual herds in Germany. In this cohort the prevalence of inflammation and necrosis in the tails, ears, claw coronary bands, heels and teats was determined using a pre-defined scoring system. Results show that already in the 1^st days of life, piglets were affected by inflammation and necrosis of the heels (80%), claw coronary bands (50%) and tail base (20%). The praevalences of these alterations in piglets were influenced by genetics (P <0.001) and age, decreasing gradually in the 2^nd week of life (P <0.001). Moreover, a correlation between tail length after tail docking and the prevalence of tail necrosis (P⩽0.04) was found. Tail and ear biting as a behavioural trait was not detected during this study. The early onset, appearance and multiple locations of clinical signs of inflammation and the positive correlation with the genetic background of the piglets may suggest an impairment of the innate immune system by infectious and non-infectious agents. This is in contrast to previously described behavioural abnormalities seen in fattening pigs. Considering the obvious reduction of animal welfare due to the described lesions, there is a need to create awareness among pig farmers and to understand the multifactorial causality involved in this inflammation and necrosis syndrome in piglets.     

An investigation into the effectiveness of compressed straw blocks in reducing abnormal behaviour in growing pigs

The provision of manipulable material to pigs is a legal requirement to ensure their good welfare. Loose straw is edible, chewable, investigable and manipulable, and has been proven to be effective in reducing abnormal behaviour. However, it is incompatible with slatted systems and therefore not a viable option for many intensive units. Thus, there is a need to identify enrichment materials that are as effective as loose straw and compatible with slatted floors. This study investigated the viability of using compressed straw blocks on a commercial farm in terms of cost and effectiveness in reducing biting relative to plastic hanging toys. A total of 880 slaughter pigs were used, among which half (n = 440, 8 groups) were provided with commercial pig toys (TOY) and the other half (n = 440, 8 groups) were provided compressed straw blocks (STRAW BLOCK). Animals were separated according to sex to investigate whether there was a higher propensity to tail and ear bite depending on sex. Pigs were monitored from weaning to slaughter, with tails being examined post scalding and any condemnations recorded at the abattoir. Throughout their life, the tails and ears of all pigs were examined individually, and behavioural observations of each pig group were conducted fortnightly. Salivary cortisol was also obtained from a subsample of pigs from each group every fortnight to monitor stress levels. The highest straw usage was recorded in the second weaner stage (39 to 67 days after weaning). Enrichment type or sex had no effect on tail lesion scores or cold carcass weight recorded at the abattoir. There was also no effect of enrichment type or sex on body, tail, or ear lesion scores during either the weaner (0 to 39 days after weaning) or finisher stage (67 to145 days after weaning). Female pigs showed more biting behaviour than males, and female pigs that were provided STRAW BLOCK performed better than those provided TOY. In both the weaner and finisher accommodation, more instances of interaction were recorded with TOY pens as enrichment than with STRAW BLOCK, but the interaction duration was not recorded. There was no effect of sex on either stage. More instances of aggression were observed with the TOY than with STRAW BLOCK in the weaner stage (P < 0.05). Enrichment type or sex had no effect on cortisol levels. Thus, relative to plastic hanging toys, compressed straw blocks provided in this manner did not benefit pig welfare.     

Investigation of the adaptor protein PLIC-2 in multiple pathways

PLIC, Protein Linking IAP (CD47) to Cytoskeleton, have long since been implicated in connecting the extracellular membrane to the intracellular cell cytoskeleton. This phenomenon is supposedly achieved by bridging a receptor protein CD47 to vimentin, an intermediate filament, which in turn regulates integrin dependent cell spreading. Since the discovery of these proteins, the molecular details of the above-mentioned interactions and the underlying complexes are yet to be characterized. Several independent studies have together emphasized PLIC/Ubiquilin’s role in the proteasomal degradation pathway. This seems to be in contrast to the purported initial discovery of PLIC as a cytoskeletal adaptor protein. In an effort to reconcile the different roles associated with the ubiquitous PLIC proteins, we tested the involvement of PLIC-2 both in the proteasomal degradation pathway and as a protein linking the cell cytoskeleton to the cytoplasmic tail of CD47. This was achieved thorough an in vitro investigation of their binding interface using a combination of biophysical techniques. Our results show that the two terminal domains of PLIC-2 interact weakly with each other, while the C-terminal UBA domain interacts strongly with ubiquitin. Interestingly, no perceptible interaction was observed for PLIC-2 with the cytoplasmic tail of CD47 questioning its role as a “PLIC” protein linking the cell membrane to the cytoskeleton.