Facultative parthenogenesis discovered for the first time in Jamaican boa (Chilabothrus subflavus) using novel microsatellite markers.

The mode of reproduction most often seen in snakes is sexual, but studies have noted facultative parthenogenesis in at least six families. Here, we provide evidence for the first observed case of facultative parthenogenesis in a captive Jamaican boa (Chilabothrus subflavus). A 7-year-old female Jamaican boa, isolated since birth, was found to have produced a litter of 15 offspring. To provide molecular DNA evidence of parthenogenesis, 13 new microsatellite loci were isolated in the species. All offspring were found to be homozygous at each locus and only possess alleles found in the dam, implicating that they were born from asexual reproduction. Several developmental abnormalities, including stillbirths and spinal deformities, were noted in the litter which may be explained by their increased level of homozygosity. To preserve genetic diversity in the captive population, research should be conducted to understand the prevalence of this mode of reproduction and to guide future management decisions of this IUCN listed Vulnerable species.

Phylogenomic Reconstruction of Sportive Lemurs (genus Lepilemur) Recovered from Mitogenomes with Inferences for Madagascar Biogeography.

The family Lepilemuridae includes 26 species of sportive lemurs, most of which were recently described. The cryptic morphological differences confounded taxonomy until recent molecular studies; however, some species' boundaries remain uncertain. To better understand the genus Lepilemur, we analyzed 35 complete mitochondrial genomes representing all recognized 26 sportive lemur taxa and estimated divergence dates. With our dataset we recovered 25 reciprocally monophyletic lineages, as well as an admixed clade containing Lepilemur mittermeieri and Lepilemur dorsalis. Using modern distribution data, an ancestral area reconstruction and an ecological vicariance analysis were performed to trace the history of diversification and to test biogeographic hypotheses. We estimated the initial split between the eastern and western Lepilemur clades to have occurred in the Miocene. Divergence of most species occurred from the Pliocene to the Pleistocene. The biogeographic patterns recovered in this study were better addressed with a combinatorial approach including climate, watersheds, and rivers. Generally, current climate and watershed hypotheses performed better for western and eastern clades, while speciation of northern clades was not adequately supported using the ecological factors incorporated in this study. Thus, multiple mechanisms likely contributed to the speciation and distribution patterns in Lepilemur.

Continuity of care experiences: A national cross-sectional survey exploring the views and experiences of Australian students and academics.

BACKGROUND: Continuity of Care Experiences (CoCEs) are a component of all entry-to-practice midwifery programs in Australia and facilitate an understanding of the central midwifery philosophy of woman-centred care and continuity of the therapeutic relationship. The aim of this research was to explore how CoCEs are viewed and experienced by students and academics across Australia. METHODS: Students enrolled in Australian midwifery programs and academics who teach into these programs were invited to participate in a cross sectional, web-based survey. Data were analysed using descriptive statistics and free text responses were analysed using content analysis. FINDINGS: Four hundred and five students and 61 academics responded to the survey. The CoCE was viewed as a positive and unique learning experience, preparing students to work in midwifery-led continuity models and developing confidence in their midwifery role. Challenges in recruitment, participation in care, and balancing the workload with other course requirements were evident in reports from students, but less understood by academics. Significant personal impact on finances, health and wellbeing of students were also reported. DISCUSSION: The value of CoCEs as an experiential learning opportunity is clear, however, many students report being challenged by elements of the CoCE within current models as they try to maintain study-work-life balance. CONCLUSION: Innovative course structure that considers and embeds the CoCE requirements within the curricula, in addition to a collective commitment from regulatory bodies, the maternity care sector and Universities to facilitate CoCEs for students may address some of the significant student impacts that are reported by this research.

Nuclear and Mitochondrial Phylogenomics of the Sifakas Reveal Cryptic Variation in the Diademed Sifaka.

The most comprehensive phylogenomic reconstruction to date was generated on all nominal taxa within the lemur genus Propithecus. Over 200 wild-caught individuals were included in this study to evaluate the intra and interspecific relationships across this genus. Ultraconserved Elements (UCEs) resulted in well-supported phylogenomic trees. Complete mitochondrial genomes (CMGs) largely agreed with the UCEs, except where a mitochondrial introgression was detected between one clade of the diademed sifaka (Propithecus diadema) and the Milne-Edwards sifaka (P. edwardsi). Additionally, the crowned (P. coronatus) and Von der Decken's (P. deckeni) sifakas belonged to a single admixed lineage from UCEs. Further sampling across these two species is warranted to determine if our sampling represents a hybrid zone. P. diadema recovered two well-supported clades, which were dated and estimated as being ancient as the split between the Perrier's (P. perrierii) and silky (P. candidus) sifakas. The reconstructed demographic history of the two clades also varied over time. We then modeled the modern ecological niches of the two cryptic P. diadema clades and found that they were significantly diverged (p < 0.01). These ecological differences result in a very limited zone of geographic overlap for the P. diadema clades (<60 km2). Niche models also revealed that the Onive River acts as a potential barrier to dispersal between P. diadema and P. edwardsi. Further taxonomic work is required on P. diadema to determine if its taxonomic status should be revised. This first genomic evaluation of the genus resolved the relationships between the taxa and the recovered cryptic diversity within one species.

Telephone triage in midwifery practice: A cross-sectional survey.

BACKGROUND: Childbearing women commonly access maternity services via the telephone. A midwife receiving these calls listens to the woman's concerns and then triages women according to their assessment. This may result in the provision of advice and instruction over the telephone or inviting the woman into the health service for further assessment. Midwives are responsible for all care and advice given to women, including via the telephone. OBJECTIVES: The purpose of this study was to explore the experiences and practices of midwives regarding their management of telephone triage. DESIGN: A cross-sectional survey. SETTING AND PARTICIPANTS: Purposive non-probabilistic sampling of currently practising midwife members of professional organisations was used to recruit participants. From this, 242 midwives responded and 230 returned valid surveys were used in data analysis. METHODS: Participant demographics, telephone triage processes, skills, educational preparation, confidence and anxiety levels, and external factors that influence midwives' management of telephone triage were collected via an on-line survey. Descriptive statistics and further analyses were conducted to explore relationships between variables. RESULTS: Eighty-three percent of midwives respond to 2-5 telephone calls per shift, with only 11.7% (n = 24) of midwives reporting that this is included in their workloads. Telephone triage is frequently managed in environments with distractions. Most midwives (84%; n = 177) report receiving no training in this skill. Confidence in performing telephone triage was reported, with higher confidence levels related to midwives' increased years of experience (p < 0.05) and age (p < 0.01). Anxiety related to managing telephone triage has been experienced by 73% (n = 151) of midwives, with this being greater in midwives with less years of experience. Anxiety is reported less by midwives in rural or remote settings compared to metropolitan or regional (p < 0.05) settings in this study. A variety of standards and aids to guide practice, and document calls are utilised in a range of ways. CONCLUSION: To the authors' knowledge, this is the first study conducted to explore midwives' practises in telephone triage. The findings suggest the need for appropriate environments to conduct telephone calls and the inclusion of telephone triage in midwifery workloads. In addition, consistent education and processes are required to reduce anxiety and support midwives provision of this service to women.

Telephone triage and midwifery: A scoping review.

BACKGROUND: Midwives use telephone triage to provide advice and support to childbearing women, and to manage access to maternity services. Telephone triage practises are important in the provision of accurate, timely and appropriate health care. Despite this, there has been very little research investigating this area of midwifery practice. AIM: To explore midwives and telephone triage practises; and to discuss the relevant findings for midwives managing telephone calls from women. METHODS: A five-stage process for conducting scoping reviews was employed. Searches of relevant databases as well as grey literature, and reference lists from included studies were carried out. FINDINGS: A total of 11 publications were included. Thematic analysis was used to identify key concepts. We grouped these key concepts into four emergent themes: purpose of telephone triage, expectations of the midwife, challenges of telephone triage, and achieving quality in telephone triage. DISCUSSION: Telephone triage from a midwifery perspective is a complex multi-faceted process influenced by many internal and external factors. Midwives face many challenges when balancing the needs of the woman, the health service, and their own workloads. Primary research in this area of practice is limited. CONCLUSION: Further research to explore midwives' perceptions of their role, investigate processes and tools midwives use, evaluate training programs, and examine outcomes of women triaged is needed.

Demonstration of enhanced endogenous fibrinolysis in thrombin activatable fibrinolysis inhibitor-deficient mice.

To investigate the importance of thrombin activatable fibrinolysis inhibitor (TAFI) in the stabilization of plasma clots, we have compared fibrinolysis in TAFI-deficient (KO) and wild-type (WT) littermate mice. TAFI-deficient mice were previously generated by targeted gene disruption. The level of TAFI activity generated in plasma from WT mice in the presence of added thrombin and thrombomodulin (activatable TAFI) is twice that of plasma from TAFI heterozygous mice (HET); no activatable TAFI is detected in TAFI KO plasma. In vitro, TAFI KO plasma clots lysed faster than WT plasma clots, and HET plasma clots lysed at an intermediate rate. The rate of clot lysis for KO mice is not changed in the presence of potato carboxypeptidase inhibitor, a specific inhibitor of TAFIa, whereas the WT and HET clot lysis rates are increased in the presence of potato carboxypeptidase inhibitor. C-terminal lysine residues are preserved on partially degraded clots from KO mice, but are absent from partially degraded WT clots. In vivo, in a batroxobin-induced pulmonary embolism model, KO mice displayed a lower retention of fibrin in the lungs than did WT mice. These results are the first demonstration of enhanced endogenous fibrinolysis in an in vivo model without the addition of exogenous thrombolytic.

Synthesis and evaluation of imidazole acetic acid inhibitors of activated thrombin-activatable fibrinolysis inhibitor as novel antithrombotics.

Thrombin-activatable fibrinolysis inhibitor (TAFI) is an important regulator of fibrinolysis, and inhibitors of this enzyme have potential use in antithrombotic and thrombolytic therapy. Appropriately substituted imidazole acetic acids such as 10j were found to be potent inhibitors of activated TAFI and selective versus the related carboxypeptidases CPA, CPN, and CPM but not CPB. Further, 10j accelerated clot lysis in vitro and was shown to be efficacious in a primate model of thrombosis.

Description of an early discharge post-acute care program: length of hospital stay, patient and carer needs and cost.

The objective of the project was to evaluate a pilot Post Acute Community Care (PACC) program for orthopaedic patients. A series of cross-sectional surveys elicited responses of patient and home carer needs and GP and hospital staff acceptability while a cost-minimisation analysis compared the average cost of the PACC program with general orthopaedic hospital care. Patients were classified according to Australian National Diagnosis Related Groups (DRGs). Average length of hospital stay in 1998/99 for PACC patients was 7.7 days compared to 12.3 for general orthopaedic patients. Only 3% of patients had an unplanned readmission to hospital. Patients and carers expressed a number of unmet needs. This study confirms the popularity of early discharge schemes with patients, and provides little evidence of adverse health outcomes or that the burden of care is shifted to carers in a way that is unacceptable for this older population.

Therapeutic hypothermia post-cardiac arrest: a clinical nurse specialist initiative in Pakistan.

PURPOSE: The purpose of this project was to assess the feasibility of an evidence-based therapeutic hypothermia protocol in adult post-cardiac arrest (CA) patients in a university hospital in Pakistan. BACKGROUND: Cardiac arrest has a deleterious effect on neurological function, and survival is associated with significant morbidity. The International Liaison Committee of Cardiopulmonary Resuscitation and the American Heart Association recommend the use of mild hypothermia in post-CA victims to mitigate brain injury caused by anoxia. In Pakistan, the survival rate in CA victims is poor. At present, there are no hospitals in the country that use the evidence-based hypothermia intervention in adult post-CA victims. DESCRIPTION: This pilot project of therapeutic hypothermia in adult post-CA patients was implemented in a university hospital in Pakistan by a clinical nurse specialist in collaboration with the cardiopulmonary resuscitation committee and the nursing leadership of the hospital. Various clinical nurse specialist competencies and roles were used to address the 3 spheres of influence: patient, nurses, and system, while executing an evidence-based hypothermia protocol. Process and outcome indicators were monitored to evaluate the effectiveness and feasibility of hypothermia intervention in this setting. OUTCOME: The hypothermia protocol was successfully implemented in 3 adult post-CA patients using cost-effective measures. All 3 patients were extubated within 72 hours after CA, and 2 patients were discharged home with good neurological outcome. CONCLUSION: Adoption of an evidence-based hypothermia protocol for adult CA patients is feasible in the intensive care setting of a university hospital in Pakistan. IMPLICATIONS: The process used in the project can serve as a road map to other hospitals in resource-limited countries such as Pakistan that are motivated to improve post-CA outcomes. This experience reveals that advanced practice nurses can be instrumental in translation of evidence into practice in a healthcare system in Pakistan.

How effectively do midwives manage the care of obese pregnant women? A cross-sectional survey of Australian midwives.

BACKGROUND: Obesity and overweight are common issues for pregnant women and their healthcare providers. Obesity in pregnancy is associated with poorer maternal and perinatal outcomes and presents particular challenges in day-to-day clinical practice. QUESTION: The aim of this study was to examine midwifery clinical practice for obese pregnant women. METHODS: We conducted a cross-sectional survey of midwives using an on-line survey distributed to members of the Australian College of Midwives. Midwives were asked about: the extent to which they provided evidence-based care; their use of a clinical guideline; their education and training and confidence to counsel obese pregnant women. Data for the questions about knowledge, clinical practice and views of education and training were summarized using descriptive statistics. Unadjusted analyses were undertaken to examine the association between use of a guideline and provision of evidence-based care and ratings of education, training and counselling. RESULTS: The survey highlighted considerable variations in practice in the care and management of obese pregnant women. Respondents' clinical knowledge and their views about education and training and counselling skills highlighted some deficits. Those using a clinical guideline were more likely to report that they 'always': tell the woman she is overweight or obese (OR 3.5; 95% CI: 1.9, 6.4); recommend a higher dose of folic acid (OR 4.6; 95% CI: 1.9, 6.4); refer to an obstetrician (OR 2.9; 95% CI: 1.2, 3.4); prepare a pregnancy plan (OR 2.0; 95% CI: 1.2, 3.3) and plan to obtain an anaesthetic referral (OR 2.6; 95% CI: 1.5, 4.3). They were also more likely to report adequate/comprehensive education and training and greater confidence to counsel obese pregnant women. CONCLUSIONS: Registered midwives need continuing professional development in communication and counselling to more effectively manage the care of obese pregnant women. The universal use of a clinical guideline may have a positive impact by helping midwives to base early care decisions on clinical evidence.

Undergraduate midwifery students' sense of belongingness in clinical practice.

BACKGROUND: Clinical placements form a large and integral part of midwifery education. While much has been written about nursing students' clinical placements, less is known about clinical experiences of undergraduate midwifery students. In nursing, belongingness has been demonstrated to be a key factor in clinical learning but little is known about this in midwifery education. OBJECTIVES: This study sought to examine undergraduate midwifery students' sense of belongingness in their clinical practice. DESIGN: A quantitative design using an online questionnaire was employed. A tool adapted by Levett-Jones (2009a), and previously used with nursing students, was utilised to examine sense of belonging in undergraduate midwifery students. PARTICIPANTS: Sixty undergraduate midwifery students from two campuses at one Australian university participated in the study. Students were drawn from a single Bachelor of Midwifery degree and a double Bachelor of Nursing/Bachelor of Midwifery degree. METHODS: On completion of a scheduled lecture, students were invited by one of the researchers to participate in the study by completing the online questionnaire and the link provided. Data were analysed using descriptive statistics. RESULTS: Midwifery students generally reported similar perceptions of belongingness with previous studies on nursing students. However, a few differences were noted that require further exploration to fully understand. CONCLUSIONS: Midwifery students experienced a sense of belonging in their clinical placements. The findings contribute to understandings of the experiences for midwifery students and provide a foundation on which to develop future clinical placement experiences.

Complete sequence and gene organization of the mitochondrial genome for Hubbard's sportive lemur (Lepilemur hubbardorum).

The complete mitochondrial DNA (mtDNA) genome of Hubbard's or Zombitse sportive lemur (Lepilemur hubbardorum) was generated by polymerase chain reaction (PCR) amplification, primer-walking sequencing and fragment cloning. Comparative analyses of Hubbard's sportive lemur were conducted with available complete mitochondrial genome sequences from eight other lemur species. The mitochondrial genome of Hubbard's sportive lemur is 16,854 base pairs (bp) and contains 13 protein-coding genes, 22 transfer RNA (tRNA) genes, two ribosomal RNA (rRNA) genes and one control region. Three rare start codons were found, in which GTG is the start codon for the ATPase 6 subunit gene (ATP), ATC for the NADH dehydrogenase (ND) 2 subunit gene, and ATT for the ND5 subunit gene. In the control region, sequence analysis found one repetitive unit between conserved sequence blocks (CSB)-1 and CSB-2 for L. hubbardorum. Comparative analysis of eight other lemur species showed different repetitive units between and outside of these two blocks. According to the phylogenetic analysis of the 12 heavy-strand encoded protein-coding genes, all nine lemur species representative of four lemuriformes families were monophyletic. This template and the newly designed primers described in this study will allow scientists to generate comparative sequences for all sportive lemurs to validate phylogenetic discrepancies in the genus Lepilemur and to evaluate evolutionary and biogeographic models.

Electrochemiluminescence assay for basic carboxypeptidases: inhibition of basic carboxypeptidases and activation of thrombin-activatable fibrinolysis inhibitor.

Carboxypeptidases catalyze the removal of the C-terminal amino acid residues in peptides and proteins and exert important biological functions. Assays for carboxypeptidase activity that rely on change of absorbance generally suffer from low sensitivity and are difficult to adapt to high-throughput screening. We have developed a sensitive, robust assay for basic carboxypeptidase activity that makes use of electrochemiluminescent (ECL) detection of reaction product. In this assay, a peptide substrate contains the epitope for antibody (G2-10) binding which is masked by a C-terminal arginine. Carboxypeptidase activity exposes the epitope, allowing the binding of ruthenylated G2-10 which is then detected using ECL. High sensitivity allowed detection limits of 1-2 pM enzyme for carboxypeptidase B and activated thrombin-activatable fibrinolysis inhibitor (TAFIa). The inhibition of several basic carboxypeptidases by commercially available inhibitors was studied. This antibody-based method can be extended to other sensitive detection techniques such as amplified luminescent proximity homogeneous assay. The high sensitivity of the assay allowed the determination of the activatable levels of TAFI in human and other animal plasma in the presence of epsilon -aminocaproic acid, an active-site inhibitor that stabilizes TAFIa. A method to isolate in situ activated TAFIa from human serum in the presence of epsilon -aminocaproic acid was also developed.

Miniaturizable homogenous time-resolved fluorescence assay for carboxypeptidase B activity.

An epitope-unmasking, homogeneous time-resolved fluorescence (HTRF) assay has been developed for measuring carboxypeptidase B (CPB) activity in a miniaturized high-throughput screening format. The enzyme substrate (biotin-RYRGLMVGGVVR-OH) is cleaved by CPB at the C terminus, causing release of the C-terminal Arg residue. The product (biotin-RYRGLMVGGVV-OH) is recognized specifically by a monoclonal antibody (G2-10) which is labeled with Eu(3+)-cryptate ([Eu(3+)]G2-10 mAb), and the complex is detected by fluorescence resonance energy transfer using streptavidin labeled with allophycocyanin ([XL665]SA). The CPB HTRF assay is readily adapted from 96- to 1536-well format as a robust (Z(')>0.5) assay for high-throughput screening.

Characterization of the inhibition of hepatitis C virus RNA replication by nonnucleosides.

The RNA-dependent RNA polymerase of hepatitis C virus (HCV) is necessary for the replication of viral RNA and thus represents an attractive target for drug development. Several structural classes of nonnucleoside inhibitors (NNIs) of HCV RNA polymerase have been described, including a promising series of benzothiadiazine compounds that efficiently block replication of HCV subgenomic replicons in tissue culture. In this work we report the selection of replicons resistant to inhibition by the benzothiadiazine class of NNIs. Four different single mutations were identified in separate clones, and all four map to the RNA polymerase gene, validating the polymerase as the antiviral target of inhibition. The mutations (M414T, C451R, G558R, and H95R) render the HCV replicons resistant to inhibition by benzothiadiazines, though the mutant replicons remain sensitive to inhibition by other nucleoside and NNIs of the HCV RNA polymerase. Additionally, cross-resistance studies and synergistic inhibition of the enzyme by combinations of a benzimidazole and a benzothiadiazine indicate the existence of nonoverlapping binding sites for these two structural classes of inhibitors.

Imidazole acetic acid TAFIa inhibitors: SAR studies centered around the basic P(1)(') group.

Structural modifications of the aminopyridine P(1)(') group of imidazole acetic acid based TAFIa inhibitors led to the discovery of the aminocyclopentyl analog 28, a 1 nM TAFIa inhibitor with CLT(50) functional activity of 14 nM but without selectivity against CPB. While not as active, aminobutyl derivative 27 provided an improved 6.7-fold selectivity for TAFIa versus CPB.