Stress resilience-associated behaviors following predator scent stress are accompanied by upregulated nucleus accumbens mGlu5 transcription in female Sprague Dawley rats.

Despite the higher prevalence of post-traumatic stress disorder (PTSD) in women, the majority of preclinical research has been conducted utilizing male subjects. We have found that male rats exposed to the predator scent 2,4,5-trimethyl-3-thiazoline (TMT) show heterogenous long-term anxiety-like behavior and conditioned fear to the TMT environment. Stress-Resilient males exhibit increased mGlu5 mRNA expression in the basolateral amygdala (BLA) and prefrontal cortex (PFC). Here we sought to determine whether the same behavioral and genetic responses would be observed in female rats exposed to TMT. Female Sprague-Dawley rats were exposed to TMT for ten minutes, while Controls were exposed to an unscented environment. Anxiety and anhedonia were assessed 7-14 days later with elevated plus maze (EPM), acoustic startle response, light-dark box, and sucrose preference test (SPT). TMT-exposed females spent less time in the EPM open arms, exhibited greater startle amplitude, and reduced sucrose intake compared to Controls. Median split analyses conducted on EPM and SPT data yielded stress-Susceptible and -Resilient phenotypes that displayed behavior in the light-dark box consistent with EPM and SPT behavior. Susceptible females displayed greater BLA mGlu5 mRNA expression than Resilient and Control rats and did not show conditioned fear, in contrast to previous results in males. Resilient females displayed greater mGlu5 mRNA expression in the nucleus accumbens. These data indicate that the predator scent stress model of PTSD produces distinct stress-Susceptible and Resilient phenotypes in female rats that are associated with changes in mGlu5 mRNA expression in several brain regions.

Is Zimbabwe ready to transition from anonymous unlinked sero-surveillance to using prevention of mother to child transmission of HIV (PMTCT) program data for HIV surveillance?: results of PMTCT utility study, 2012.

BACKGROUND: Prevention of mother-to-child transmission of HIV (PMTCT) programs collect socio-demographic and HIV testing information similar to that collected by unlinked anonymous testing sero-surveillance (UAT) in antenatal settings. Zimbabwe evaluated the utility of PMTCT data in replacing UAT. METHODS: A UAT dataset was created by capturing socio-demographic, testing practices from the woman's booking-card and testing remnant blood at a laboratory from 1 June to 30 September 2012. PMTCT data were collected retrospectively from ANC registers. UAT and PMTCT data were linked by bar-code labels that were temporarily affixed to the ANC register. A questionnaire was used to obtain facility-level data at 53 sites. RESULTS: Pooled HIV prevalence was 15.8 % (95 % CI 15.3-16.4) among 17,349 women sampled by UAT, and 16.3 % (95 % CI 15.8 %-16.9 %) among 17,150 women in PMTCT datasets for 53 sites. Pooled national percent-positive agreement (PPA) was 91.2 %, and percent-negative agreement (PNA) was 98.7 % for 16,782 women with matched UAT and PMTCT data. Based on UAT methods, overall median prevalence was 12.9 % (Range 4.0 %-19.4 %) among acceptors and refusers of HIV test in PMTCT compared to 12.5 % ((Range 3.4 %-19.5 %) among acceptors in ANC registers. There were variations in prevalence by site. CONCLUSION: Although, there is no statistical difference between pooled HIV prevalence in UAT compared to PMTCT program, the overall PPA of 91.2 % and PNA of 98.7 % fall below World Health Organisation (WHO) benchmarks of 97.6 % and 99.6 % respectively. Zimbabwe will need to strengthen quality assurance (QA) of rapid HIV testing and data collection practices. Sites with good performance should be prioritised for transitioning.

Doing the right thing at the right time.

OBJECTIVE: To identify factors that clinicians consider when a patient is dying, enabling implementation of the Liverpool Care Pathway. BACKGROUND: In order to improve the care of the dying patient and their families it is helpful to implement the Liverpool Care Pathway for the dying. It therefore is necessary to identify the dying patient in a timely fashion. METHOD: A phenomenological study using semi-structured interviews (n=five nurses and five doctors) conducted on a hospice inpatient unit. FINDINGS: There was a prominent theme of anxiety about getting the timing of diagnosing dying right, with an emphasis how the dying patient and their families would cope if this were wrong. The main factors identified were: support for decision making, understanding the patient's journey and concern that the care given is appropriate. CONCLUSIONS AND IMPLICATIONS FOR NURSING PRACTICE: All clinicians interviewed for this study had concerns about increasing the patient's/carers' distress if the Liverpool Care Pathway implementation was mistimed. There is a risk that clinicians are avoiding difficult conversations with families and there may be a lack of understanding around the reasons for use of the Liverpool Care Pathway. Specific communications training may help clinicians in this role.

The history of neurosurgery in Memphis: the Semmes-Murphey Clinic and the Department of Neurosurgery at the University of Tennessee College of Medicine.

Neurological surgery was defined as a separate surgical specialty by Harvey Cushing and a few other surgeons, most of whom were trained and influenced by Cushing. One of these, Raphael Eustace Semmes, became the first neurosurgeon in Memphis, Tennessee, in 1912. After World War II, Semmes and his first associate, Francis Murphey, incorporated the Semmes-Murphey Clinic, which has been primarily responsible for the growth of the Department of Neurosurgery at the University of Tennessee Health Science Center in Memphis, as well as the development of select neurosurgical subspecialties in Memphis area hospitals.

The function of factor XI in tissue factor-initiated thrombin generation.

The influence of plasma and platelet factor (F)XI on thrombin generation initiated with 10 pm tissue factor (TF) in a synthetic coagulation model was evaluated in the presence of either 2 x 108 mL-1 platelets or the equivalent (2 microm) phospholipids. In either system, with all proteins present at physiological concentrations, FXI (30 nm) had no effect on thrombin generation. With phospholipids in the absence of FXI, an increase in vitamin K-dependent proteins (VKDP) (up to 500%) significantly prolonged the initiation phase of thrombin generation and decreased maximum thrombin levels. The inhibition was principally caused by the elevated prothrombin and FIX concentrations. When 30 nm FXI was added with elevated VKDP and phospholipids, the initiation phase was decreased and the maximum thrombin levels generated substantially increased. In experiments with platelets (with and without plasma FXI), an increase in VKDP had little effect on the initiation phase of thrombin generation. These data indicate that (i) FXI has no effect on thrombin generation at 10 pm TF and physiological concentrations of VKDP; (ii) platelets and plasma FXI are able to compensate for the inhibitory effects of elevated VKDP.

Clonal restriction of platelet-associated anti-GPIIb/IIIa autoantibodies in patients with chronic ITP.

Chronic immune thrombocytopenic purpura is due to platelet destruction induced by autoantibodies against platelet surface antigens. Prior studies show that some serum autoantibodies are light-chain restricted, suggesting a clonal origin. Since plasma and platelet-associated antibody from the same patient may bind to different epitopes. it is important to evaluate the clonality of platelet-associated antibody. Platelet-associated autoantibodies from 28 ITP patients were studied. Of 23 platelet-associated antibodies tested directly, 16 showed significant light chain restriction (7 complete and 9 partial) when compared to plasma IgG light chain distribution. Similarly, 9 of 12 platelet-associated antibody eluates were light chain restricted, 5 complete and 4 partial. In all cases where platelet-associated antibody and antibody eluate from the same patient were studied, the results were concordant. We conclude that a significant proportion of platelet-associated antibodies from ITP patients show apparent clonality, as evaluated by light chain restriction. These results are consistent with other studies in ITP suggesting a limited antigenic repertoire.

Autoantibodies to alpha(IIb)beta(3) in patients with chronic immune thrombocytopenic purpura bind primarily to epitopes on alpha(IIb).

Chronic immune thrombocytopenic purpura (ITP) is an autoimmune disease caused by platelet destruction resulting from autoantibodies against platelet surface proteins, particularly platelet glycoprotein IIb/IIIa (alpha(IIb)beta(3)). To localize the auto-epitopes on platelet alpha(IIb)beta(3), the binding of autoantibodies to Chinese hamster ovary (CHO) cells expressing either alpha(IIb)beta(3) or alpha(v)beta(3) was studied. Thirteen of 14 ITP autoantibodies bound only to CHO cells expressing alpha(IIb)beta(3). Because these 2 integrins have the same beta chain (beta(3)), these results show that most epitopes in chronic ITP are dependent on the presence of glycoprotein alpha(IIb.) (Blood. 2001;97:2171-2172)

The Shc-related adaptor protein, Sck, forms a complex with the vascular-endothelial-growth-factor receptor KDR in transfected cells.

Despite much progress in recent years, the precise signalling events triggered by the vascular-endothelial-growth-factor (VEGF) receptors, fms-like tyrosine kinase (Flt1) and kinase insert domain-containing receptor (KDR), are incompletely defined. Results obtained when Flt1 and KDR are individually expressed in fibroblasts or porcine aortic endothelial cells have not been entirely consistent with those observed in other endothelial cells expressing both receptors endogenously. It has also been difficult to demonstrate VEGF-induced phosphorylation of Flt1, which has led to speculation that KDR may be the more important receptor for the mitogenic action of VEGF on endothelial cells. In an attempt to identify physiologically important effectors which bind to KDR, we have screened a yeast two-hybrid mouse embryo library with the cytoplasmic domain of KDR. Here we describe the identification of the adaptor protein, Shc-like protein (Sck), as a binding partner for KDR. We demonstrate that this interaction requires phosphorylation of KDR, and identify the binding site for the Src-homology 2 (SH2) domain as tyrosine-1175 of KDR. We have also shown that the SH2 domain of Sck, but not that of Src-homology collagen protein (Shc), can precipitate phosphorylated KDR from VEGF-stimulated porcine aortic endothelial cells expressing KDR, and that an N-terminally truncated Sck protein can associate with KDR, in a phosphorylation-dependent fashion, when co-expressed in human embryonic kidney 293 cells. Furthermore, we demonstrate that in the two-hybrid assay, both Shc and Sck SH2 domains can associate with the related receptor Flt1.

Identification of three genes expressed primarily during development in Physarum polycephalum.

During the life cycle of Physarum polycephalum, uninucleate amoebae develop into multinucleate syncytial plasmodia. These two cell types differ greatly in cellular organisation, behaviour and gene expression. Classical genetic analysis has identified the mating-type gene, matA, as the key gene controlling the initiation of plasmodium development, but nothing is known about the molecular events controlled by matA. In order to identify genes involved in regulating plasmodium formation, we constructed a subtracted cDNA library from cells undergoing development. Three genes that have their highest levels of expression during plasmodium development were identified: redA, redB (regulated in development) and mynD (myosin). Both redA and redB are single-copy genes and are not members of gene families. Although redA has no significant sequence similarities to known genes, redB has sequence similarity to invertebrate sarcoplasmic calcium-binding proteins. The mynD gene is closely related to type II myosin heavy-chain genes from many organisms and is one of a family of type II myosin genes in P. polycephalum. Our results indicate that many more red genes remain to be identified, some of which may play key roles in controlling plasmodium formation.

Disturbances of antagonistic neck innervation in patients with cerebellar deficits.

Fast 60 degree head rotations of nine patients with cerebellar deficits were analysed and compared with those of nine normal subjects. The surface EMG activity from both Splenii capitis muscles were recorded. The triphasic pattern of reciprocal innervated neck muscles with regard to the duration, amplitude and onset of the pulses were analysed together with the dynamic features of head rotation, i.e. position and acceleration profiles. The deviation of the onset of the antagonistic (B) pulse of the EMG-pattern flow was substantially increased. In most cases the onset of the B-pulse was delayed, less premature and sporadic regular onsets occurred. The number of co-contractions and multiple antagonistic pulses was significantly increased. Half of the movements of patients were found to be dysmetric, with an irregular flow of position and acceleration functions. From these, mainly hypermetric movements occurred. The number of irregular pulse patterns was higher than the number of dysmetrias. In this context cocontractions, multiple antagonistic pulses and cortical control can be discussed as strategies of the cerebellar patients to improve their dynamic output. Our experiments support the notion of the cerebellum playing an important part in the temporal integration of an antagonistically innervated movement. The measurement of electromyographic burst patterns can be used as a clinical tool to demonstrate an insufficient timing of the activities of muscles involved.

G1-phase arrest is not a prerequisite for encystment in Physarum.

Amoebae of the Myxomycete Physarum polycephalum form resistant, walled cysts when the food bacteria in a culture have been consumed. No G1 phase has been detected in the vegetative amoebal cell cycle, most of which comprises the G2 phase. Mature cysts are also in G2, but it has been reported that a G1 phase of roughly 24 h, followed by an S phase, is obligatory prior to encystment. We used flow cytometry to determine the distribution of DNA contents in amoebal cultures at intervals during vegetative growth and encystment. In all cultures, the cells were predominantly in G2 phase, and the percentage of cells with G1 DNA content remained very low. We conclude that an extended G1 phase of 24 h did not occur in our cultures and cannot be a prerequisite for encystment.

Gracilis or semitendinosus myopathy in 18 dogs.

The clinical findings in 18 dogs with gracilis (n = 17) or semitendinosus (n = 1) myopathy are described. Each dog had a similar hind-limb gait abnormality characterized by a shortened stride with a rapid, elastic medial rotation of the paw, internal rotation of the hock and external rotation of the calcaneus [corrected] and internal rotation of the stifle during the mid-to-late swing phase of the stride. Medical management prior to or in lieu of surgery was attempted (n = 8) with no apparent response. Fifteen dogs had one or multiple surgical procedures. Although transection, partial excision, or complete resection of the affected muscle resulted in resolution of lameness following surgery, lameness recurred six weeks to five months (mean, 2.5 months; median, two months) following surgery. Adjunctive medical treatment did not prevent recurrence. Variable replacement of the affected muscle with fibrous connective tissue (predominantly along the caudolateral border of the muscle) was evident grossly, and replacement of myofibers with fibrous connective tissue was confirmed histologically. A definitive etiology could not be established.

Characterization of npf mutants identifying developmental genes in Physarum.

In Physarum polycephalum, uninucleate haploid amoebae develop into macroscopic multinucleate plasmodia. Wild-type, sexual development is triggered when two amoebae carrying different alleles of matA fuse to form a zygote which develops into a diploid plasmodium. Mutations in the matA genetic region give rise to apogamic strains in which a single haploid amoeba can develop into a haploid plasmodium. An essential stage in both sexual and apogamic plasmodium formation is an extended cell cycle in uninucleate cells, which ends with the formation of a binucleate cell by mitosis without cytokinesis. Using a 'brute force' screening method, we have isolated mutants blocked in apogamic plasmodium development. Genetic analysis showed that the mutations we have identified were unlinked to matA, unlike mutations previously identified following an enrichment step. Most of the loci revealed by our screen were represented by only one allele, indicating that further screening should lead to the identification of additional genes required for plasmodium development. Phenotypic analysis showed that different mutants were blocked at different stages of plasmodium formation. Some of the mutations blocking apogamic development at an early stage, close to the start of the long cell cycle, failed to block sexual development in zygotes homozygous for the mutation. Since the two modes of plasmodium formation differ only in the initiation of development, these mutations presumably interfere with the initiation process. In the remaining mutants, in which both sexual and apogamic development were blocked, development first became abnormal towards the end of the long cell cycle. This suggested that the wild-type gene products were required by this time and was consistent with previous evidence that many changes in cellular organization and gene expression occur during the long cell cycle. Each of these mutants showed a different terminal phenotype and some aspects of plasmodium development occurred normally although others were blocked, suggesting that development involves multiple pathways rather than a dependent sequence of events. Phenotypic analysis of double mutants supported this conclusion and also revealed epistatic interactions, presumably due to blocks in the same pathway. In several of the mutants, terminally differentiated cells died by an apoptosis-like mechanism; since this was never observed in vegetative cells, it was presumably triggered by the failure of development. Phenotypic analyses of additional mutants will extend our understanding of the pathways involved in plasmodium development.

Parameter identification of a neurological control model for the pathological head movements of cerebellar patients.

The objective of this research is to explore the role of the cerebellum in the human motor control system. The present study quantitatively compares the neurological control signals effecting fast, horizontal head rotations in normal subjects to those in patients with a cerebellar lesion. The method involves the use of a computer simulation model for one degree-of-freedom movements. A method for unconstrained global optimization, first proposed by Hans Bremermann (1970), is used to identify the timing and magnitudes of the input neurological control signals to the model, which are compared to recorded electromyograms (EMGs). Experimentally recorded kinematics from cerebellar patients and from normal subjects were used to drive the parameter search. These simulations found that cerebellar patients' neurological control signals were altered with respect to those of normal subjects, and suggest that the electromyographic activity of cerebellar patients may comprise at least five bursts of activity whereas normal subjects typically exhibit only three. The results are discussed with respect to the hypothesis that the cerebellum may be involved in both the timing and magnitudes of the neurological control signals effecting voluntary movement.