State of pediatric liver transplantation in the United States and achieving zero wait list mortality with ideal outcomes: A statement from the Starzl Network for Excellence in Pediatric Transplant Surgeon's Working Group.

BACKGROUND: Liver transplant is a life-saving therapy that can restore quality life for several pediatric liver diseases. However, it is not available to all children who need one. Expertise in medical and surgical management is heterogeneous, and allocation policies are not optimally serving children. Technical variant grafts from both living and deceased donors are underutilized. METHODS: Several national efforts in pediatric liver transplant to improve access to and outcomes from liver transplant for children have been instituted and include adjustments to allocation policies, UNOS-sponsored collaborative improvement projects, and the emergence of national learning networks to study ongoing challenges in the field the Surgical Working group of the Starzl Network for Excellence in Pediatric Transplantation (SNEPT) discusses key issues and proposes potential solutions to eliminate the persistent wait list mortality that pediatric patients face. RESULTS: A discussion of the factors impacting pediatric patients' access to liver transplant is undertaken, along with a proposal of several measures to ensure equitable access to life-saving liver transplant. CONCLUSIONS: Pediatric liver transplant wait list mortality can and should be eliminated. Several measures, including collaborative efforts among centers, could be leveraged to acheive this goal.

Perception and reaction of Nanyang Technological University (NTU) researchers to different forms of research integrity education modality.

BACKGROUND: Research and academic institutions use various delivery channels to deliver Research Integrity (RI) education in their communities. Yet there is no consensus on the best delivery method and the effectiveness of these channels in inculcating a positive RI culture varies across institutions. Hence, this study aimed to understand the preferences of the research community in Nanyang Technological University (NTU), Singapore. METHODS: An online survey was conducted on NTU research community to understand their experience with, and preference for each RI education mode offered in NTU. The RI education modes surveyed in the general ranking question are Data Management Plan (DMP) workshops, Epigeum e-Learning, Compass e-newsletter (email), and NTU policy on Research Integrity and Responsible Conduct of Research. There were 242 responses, comprising 50% research students, 32.2% research staff and 17.8% faculty members. Non-parametric statistical techniques were used to analyse preferences across different RI education modes and within sub-groups (i.e., fields, age, native language, roles in research community). RESULTS: More than 92% of respondents subscribed to the importance of RI education, but with different preferences for education modes. With respect to RI education in NTU, Compass e-newsletters were ranked the lowest (p < 0.05). Most felt that they were too wordy and unengaging, making it difficult to absorb information. Similarly, Epigeum e-Learning (p < 0.05) and 'policy' (p < 0.05) were found to be too lengthy in presentation. The compulsory NTU RI education modes (Epigeum e-learning and 'policy') enjoyed higher participation rates of 70-80% compared with 32-37% for the self-regulated modes (DMP workshop and e-newsletter). This suggests that regulatory mechanisms are still necessary to promote participation in RI education, and thus, core RI education content should be made compulsory in research/academic institutions. Although Epigeum is a compulsory course, some may not have participated in the programme due to technical issues or they might have forgotten to participate in the programme within the permissible timeframe. For all four RI education modes in NTU, the lack of awareness was among the top cited reasons for not participating. CONCLUSIONS: Most NTU researchers perceived RI education positively although they may have reservations for some approaches. Conversely, e-Learning is favored over all the other modes except for the mode of Policy. Findings from this study are useful for improving the design of RI education strategies to be more appealing to the research community by enhancing user experience in terms of user-friendliness, relevance to specialisation, providing concise information and better presentation of materials For institutions with similar modes of RI education as NTU, these results may be relevant in improving participation rates and presentation of RI education modes, such as the use of infographics and more concise information.

Is lower dose of intramuscular dexamethasone injection beneficial in reducing neonatal respiratory morbidity for elective caesarean section deliveries at 37 to 38 weeks? An observational study.

INTRODUCTION: The use of intramuscular (IM) dexamethasone injections before an elective caesarean delivery at term has been shown in multiple randomized controlled trials to reduce the rates of transient tachypnoea of the newborn, and admission to neonatal care. Recent studies have suggested that a complete course of IM steroids can be associated with long term harmful consequences to the infants born, and there have been studies suggesting that a lower dose of IM corticosteroids can be beneficial. Therefore, we aim to establish if halving the dose of dexamethasone to 12mg can demonstrate any benefit for term elective caesarean section deliveries whilst minimizing foetal exposure. METHODS: An observational controlled study comparing neonatal respiratory morbidities before and after the single dose 12mg dexamethasone was implemented in our obstetrics and gynaecology centre for term elective caesarean section deliveries. We included singleton pregnancies from 37+0 to 38+6 weeks undergoing elective caesarean section into our study. A total of 674 patients fulfilled the inclusion criteria and were recruited. We compared the rates and duration of admission to neonatal intensive care unit, the need for mechanical ventilation and the rate of transient tachypnoea of the newborn in the first half of 2019 without IM dexamethasone injections against the second half of the year when a single dose IM dexamethasone was given. RESULTS: IM dexamethasone injection did not show any significant benefit with regards to reducing the admission to neonatal care (OR 0.97, p- value 0.69), admission to neonatal intensive care unit (OR 0.91, p- value 0.80), the need for mechanical ventilation (OR 0.98, p- value 0.95), and the incidence of transient tachypnoea of the newborn (OR1.01, p- value 0.96). There was also no significant difference for the duration of admission in the neonatal intensive care unit for both groups (p- value 0.17). CONCLUSIONS: This study showed that there was no significant benefit gained from the lower dose antenatal corticosteroids for term elective caesarean section deliveries and considering that there have been long term harmful consequences demonstrated from the higher dose of antenatal corticosteroids at term, this practice should therefore be discontinued until a larger study is done to refute these findings. The use of such dexamethasone should only be a viable option in a research setting.

A Cholesterol-Dependent Endocytic Mechanism Generates Midbody Tubules During Cytokinesis.

Cytokinesis is the final stage of cell division and produces two independent daughter cells. Vesicles derived from internal membrane stores, such as the Golgi, lysosomes, and early and recycling endosomes accumulate at the intracellular bridge (ICB) during cytokinesis. Here, we use electron tomography to show that many ICB vesicles are not independent but connected, forming a newly described ICB vesicular structure - narrow tubules that are often branched. These 'midbody tubules' labelled with horseradish peroxidase (HRP) within 10 min after addition to the surrounding medium demonstrating that they are derived from endocytosis. HRP-labelled vesicles and tubules were observed at the rim of the ICB after only 1 min, suggesting that midbody tubules are likely to be generated by local endocytosis occurring at the ICB rim. Indeed, at least one tubule was open to the extracellular space, indicative of a local origin within the ICB. Inhibition of cholesterol-dependent endocytosis by exposure to methyl-ß-cyclodextrin and filipin reduced formation of HRP-labelled midbody tubules, and induced multinucleation following ICB formation. In contrast, dynamin inhibitors, which block clathrin-mediated endocytosis, induced multinucleation but had no effect on the formation of HRP-labelled midbody tubules. Therefore, our data reveal the existence of a cholesterol-dependent endocytic pathway occurring locally at the ICB, which contributes to the accumulation of vesicles and tubules that contribute to the completion of cytokinesis.

Structure of RNA 3'-phosphate cyclase bound to substrate RNA.

RNA 3'-phosphate cyclase (RtcA) catalyzes the ATP-dependent cyclization of a 3'-phosphate to form a 2',3'-cyclic phosphate at RNA termini. Cyclization proceeds through RtcA-AMP and RNA(3')pp(5')A covalent intermediates, which are analogous to intermediates formed during catalysis by the tRNA ligase RtcB. Here we present a crystal structure of Pyrococcus horikoshii RtcA in complex with a 3'-phosphate terminated RNA and adenosine in the AMP-binding pocket. Our data reveal that RtcA recognizes substrate RNA by ensuring that the terminal 3'-phosphate makes a large contribution to RNA binding. Furthermore, the RNA 3'-phosphate is poised for in-line attack on the P-N bond that links the phosphorous atom of AMP to N(ε) of His307. Thus, we provide the first insights into RNA 3'-phosphate termini recognition and the mechanism of 3'-phosphate activation by an Rtc enzyme.

A tRNA splicing operon: Archease endows RtcB with dual GTP/ATP cofactor specificity and accelerates RNA ligation.

Archease is a 16-kDa protein that is conserved in all three domains of life. In diverse bacteria and archaea, the genes encoding Archease and the tRNA ligase RtcB are localized into an operon. Here we provide a rationale for this operon organization by showing that Archease and RtcB from Pyrococcus horikoshii function in tandem, with Archease altering the catalytic properties of the RNA ligase. RtcB catalyzes the GTP and Mn(II)-dependent joining of either 2',3'-cyclic phosphate or 3'-phosphate termini to 5'-hydroxyl termini. We find that catalytic concentrations of Archease are sufficient to activate RtcB, and that Archease accelerates both the RNA 3'-P guanylylation and ligation steps. In addition, we show that Archease can alter the NTP specificity of RtcB such that ATP, dGTP or ITP is used efficiently. Moreover, RtcB variants that have inactivating substitutions in the guanine-binding pocket can be rescued by the addition of Archease. We also present a 1.4 Å-resolution crystal structure of P. horikoshii Archease that reveals a metal-binding site consisting of conserved carboxylates located at the protein tip. Substitution of the Archease metal-binding residues drastically reduced Archease-dependent activation of RtcB. Thus, evolution has sought to co-express archease and rtcB by creating a tRNA splicing operon.

Telomere dysfunction impairs DNA repair and enhances sensitivity to ionizing radiation.

Telomeres are specialized nucleoprotein complexes that serve as protective caps of linear eukaryotic chromosomes. Loss of telomere function is associated with rampant genetic instability and loss of cellular viability and renewal potential. The telomere also participates in processes of chromosomal repair, as evidenced by the 'capture' or de novo synthesis of telomere repeats at double-stranded breaks and by the capacity of yeast telomeres to serve as repositories of essential components of the DNA repair machinery, particularly those involved in non-homologous end-joining (NHEJ). Here we used the telomerase-deficient mouse, null for the essential telomerase RNA gene (Terc), to assess the role of telomerase and telomere function on the cellular and organismal response to ionizing radiation. Although the loss of telomerase activity per se had no discernable impact on the response to ionizing radiation, the emergence of telomere dysfunction in late-generation Terc-/- mice imparted a radiosensitivity syndrome associated with accelerated mortality. On the cellular level, the gastrointestinal crypt stem cells and primary thymocytes showed increased rates of apoptosis, and mouse embryonic fibroblasts (MEFs) showed diminished dose-dependent clonogenic survival. The radiosensitivity of telomere dysfunctional cells correlated with delayed DNA break repair kinetics, persistent chromosomal breaks and cytogenetic profiles characterized by complex chromosomal aberrations and massive fragmentation. Our findings establish a intimate relationship between functionally intact telomeres and the genomic, cellular and organismal response to ionizing radiation.

Australian immunisation registers: established foundations and opportunities for improvement.

The National Immunisation Program Schedule in Australia is formulated and funded nationally under the population-wide Medicare system. The policy is implemented by the eight state and territory jurisdictions. The national immunisation registers consist of the Australian Childhood Immunisation Register (ACIR), and, more recently, the National Human Papillomavirus (HPV) Vaccination Program Register. Moreover, a variety of jurisdiction-based registers and primary care practice software systems exist, which interact with the national registers. General practitioners can obtain reports listing patients under seven years attending their practice and recorded as 'not fully immunised', and immunisation coverage rates for their practice linked to government incentives through Medicare. A 2011 report documents national coverage of 91.8% fully immunised at 12 months, and 92.6% at 24 months. The HPV register provides information on vaccination coverage with the potential to link with a register of cervical cancer screening results. Limitations of current national register include inability to easily access immunisation histories beyond seven years of age, and issues of underreporting and timeliness, which impact significantly the immunisation coverage estimates. The linkage of these registers with healthcare outcome data will further enhance public health outcomes by enabling rapid, population-level vaccine safety and effectiveness investigations in a nation with a track record as an 'early adopter' of new childhood vaccines.

Anticardiolipin antibodies and lupus anticoagulants among women undergoing in vitro fertilization in Jamaica.

OBJECTIVE: The aims of this study were to evaluate the effect of anticardiolipin antibody (aCL) and lupus anticoagulant (LA) on the outcome of the in vitro ferlitization (IVF) cycles and to determine the prevalence of these antibodies in infertile women seeking IVF in Jamaica. METHODS: A retrospective cohort study was performed to determine if screening patients for aCL and LA had any significant impact on the outcome of the IVF process. Each patients hospital record, between March 2000 and March 2010, was collected and the relevant data extracted. RESULTS: The prevalence of aCL in this cohort of Jamaican women was moderate/high positive 3.88%, low positive 0.68% and those with negative aCL results 95.4%. The prevalence of women who were LA positive was 4.1% and 0.9% of the women were positive for both LA and aCL. Of the patients who were LA and/or aCL positive, eight out of 30 patients (26.7%) had a positive pregnancy test in comparison to 61 out of 181 patients (33.7%) who were LA and/or aCL negative (p = 0.5787). CONCLUSION: The prevalence of positive aCL and/or LA in infertile women seeking IVF in Jamaica is 7.76%. The presence of these antibodies did not affect the pregnancy rate of these women nor did it demonstrate an increased risk for IVF cycle cancellation or ovarian hyperstimulation syndrome. Screening women undergoing IVF for these antibodies is not justified.

Whole recombinant yeast vaccine induces antitumor immunity and improves survival in a genetically engineered mouse model of melanoma.

Malignant melanoma is one of the deadliest forms of skin cancer and its incidence is expected to rise over the next two decades. At present, there are no effective therapies for advanced melanoma. We have previously shown that administration of whole recombinant yeast expressing human MART-1 (hMART-IT) induces protective antimelanoma immunity in a B16F10 transplantable mouse model. In this study, we examine the effectiveness of the hMART-IT vaccine in a congenic strain of genetically engineered mouse model of melanoma, which recapitulates both the underlying genetics and the proper tumor microenvironment of naturally occurring melanoma. Subcutaneous administration of hMART-IT induced cytotoxicity against melanoma cells and antigen-specific production of Th1-specific cytokines by splenocytes. Weekly administration of hMART-IT significantly delayed the development of melanoma and prolonged the survival of mice compared with controls. Although histological analysis demonstrated diffuse infiltration of CD4(+) T cells and CD8(+) T cells, no reduction of regulatory T cells was observed, suggesting that hMART-IT cannot prevent immunotolerance in the tumor microenvironment. This study provides a proof of concept that genetically engineered mouse models lend valuable insights into immunotherapeutics being tested in the preclinical setting.

The influence of nitric oxide synthase 2 on cutaneous wound angiogenesis.

BACKGROUND: Inducible nitric oxide synthase (nitric oxide synthase 2, NOS 2) inhibition significantly suppresses chronically ischaemic skin flap survival, possibly because of reduced angiogenesis. OBJECTIVES: To investigate the effect of genetic NOS 2 inhibition on cutaneous wound angiogenesis in two in vivo murine models. The impact of NOS 2 manipulation on vascular endothelial growth factor (VEGF)-A stimulated and fibroblast growth factor (FGF)-2 stimulated angiogenesis was also investigated in the Matrigel(®) plug assay. METHODS: (i) Matrigel plugs/incisional wounds: two groups of NOS 2-/- mice and two groups of wild-type (WT) mice had bilateral Matrigel plugs containing 500 ng mL(-1) VEGF-A or 1000 ng mL(-1) FGF-2 injected subcutaneously in the abdomen. A 2·5 cm long dorsal incisional skin wound was created and sutured closed in the same animals. Wounds and plugs were explored at 7 or 12 days. (ii) Excisional wounds: dorsal 0·5 × 1·0 cm excisional skin wounds were created in four groups (two NOS 2-/- and two WT) and explored at 7 or 14 days. Wounds and Matrigel plugs were examined histologically and morphometrically for determination of percentage vascular volume (PVV). RESULTS: The PVV in NOS 2-/- incisional wounds and excisional wounds was significantly less than in WT wounds (P = 0·05 and P < 0·001, respectively). The PVV was significantly less in VEGF-A stimulated Matrigel plugs compared with FGF-2 stimulated plugs in NOS 2-/- mice (P < 0·01), but not in WT mice. CONCLUSIONS: NOS 2 is significantly involved in angiogenic signalling in healing skin wounds, particularly within the first 7 days. However, Matrigel plug vascularization suggests that the role of NOS 2 in angiogenesis is related to VEGF-A but not FGF-2 stimulated angiogenesis.

Propofol sedation in patients undergoing colonoscopy in Jamaica.

BACKGROUND: Propofol sedation is increasingly used for colonoscopy and may be associated with increased satisfaction and efficiency in diagnostic and therapeutic endoscopy. However propofol has a relatively narrow therapeutic window as it frequently produces deep sedation, and can precipitate respiratory depression. AIM: To determine the efficacy, safety and patient satisfaction with propofol sedation in patients undergoing colonoscopy at the University Hospital of the West Indies (UHWI). METHODS: Patients undergoing outpatient colonoscopy at the UHWI who were sedated with propofol were studied. Boluses of 10 - 20 mg of propofol at intervals of 2 - 5 minutes, as needed for adequate sedation, were administered after initial induction. Continuous monitoring of the pulse rate, and oxygen saturation were performed and the blood pressure checked every 2 - 5 minutes. All patients received supplemental oxygen (4 L/min). The following observations were recorded: the endoscopist recorded the ease of the procedure, the anaesthetist recorded the comfort of the patient throughout the procedure and at the time of discharge, and the patient stated the degree of satisfaction with the procedure. Any unusual events were recorded. RESULTS: Sixty consecutive patients sedated with propofol were studied. There were 28 (46.7%) males, with a mean age of 58.3 years and 32 (53.3%) females, with mean age of 59.5 years. Most were normal healthy patients (56.6%). Comorbid illnesses were present in 43.4%, with hypertension being most common (23.3%). All patients were classified as ASA class 1 and 2. The average dose of propofol used was 180 mg (range 50 - 355 mg). The mean duration of colonoscopy was 19.5 minutes. The mean recovery period (able to stand) was 29.6 minutes. There were no documented cases of significant hypotension, bradycardia, or hypoxaemia during the procedure. Transient apnoeic episodes during the initial stages of sedation occurred in 12 (20%) patients. The majority of patients (91.7%) rated the experience as being extremely good or excellent. The majority could not recall the actual colonoscopy and there were minimal subjective reports of nausea or discomfort during the procedure. CONCLUSIONS: Propofol sedation was associated with quick recovery and excellent satisfaction by patients and is a suitable alternative for sedation for colonoscopy in Jamaica.

Pre- and Post-operative determinants of transplantation-free survival after Fontan. The Australia and New Zealand experience.

BACKGROUND: This review identifies the predictors of late mortality and heart transplantation that remain relevant in the contemporary population of patients with a Fontan circulation, focusing on the potential impact of post-Fontan morbidities on the late outlook of these patients. METHODS AND RESULTS: A total of 1561 patients who had survived the Fontan operation in Australia or New Zealand from 1975 to 2018 were included in this review. Over a median duration of 11.4 years, there was a total of 117 deaths (7%) and 32 heart transplantations (2%). Freedom from death and heart transplantation at 10, 20 and 35 years post Fontan surgery were 94% (95% CI 93-95%), 87% (95 %CI 85-90%) and 66% (95 %CI 57-78%) respectively. Being male, having an atriopulmonary Fontan, pre-Fontan atrioventricular valve intervention, or prolonged pleural effusions post Fontan were predictive of late death or heart transplantation. However, time-dependent variables such as the development of atrial arrhythmia, protein/losing enteropathy or late ventricular dysfunction were stronger predictors of the same outcome. Patients who developed a time-dependent risk factor had a freedom from death and heart transplantation rate of 54% (95 %CI 43-66) at 15 years and 44% (95 %CI 33-57) at 25 years post Fontan. However, 95% (95 %CI 91-99) of patients without any of the identified risk factors were free from death or heart transplantation rate at 25 years post Fontan. CONCLUSION: In conclusion, the occurrence of post-operative complications such as PLE, arrhythmia and ventricular dysfunction will likely precede the late demise of these patients.

A reconfigurable optofluidic Michelson interferometer using tunable droplet grating.

This paper presents a novel optofluidic Michelson interferometer based on droplet microfluidics used to create a droplet grating. The droplet grating is formed by a stream of plugs in the microchannel with constant refractive index variation. It has a real-time tunability in the grating period through varying the flow rates of the liquids and index variation via different combinations of liquids. The optofluidic Michelson interferometer is highly sensitive and is suitable for the measurement of biomedical and biochemical buffer solutions. The experimental results show that it has a sensitivity of 66.7 nm per refractive index unit (RIU) and a detection range of 0.086 RIU.

Failure of clonal deletion in neonatally thymectomized mice: tolerance is preserved through clonal anergy.

Self-tolerance is achieved in part through intrathymic deletion of self-reactive T cells. The necessity of the thymus for this process is suggested by the development of autoimmune diseases in neonatally thymectomized (neoTx) mice and by the failure of clonal deletion in nude mice. Indeed, the present study demonstrates that neonatal thymectomy on day 3 after birth results in the failure of clonal deletion of V beta 11+ T cells in BALB/c mice and V beta 5+ and V beta 6+ T cells in DBA/2 mice. However, these potentially autoreactive cells are nonfunctional as measured by proliferation and lymphokine production after stimulation with appropriate anti-V beta mAbs or stimulator cells. It appears that this induction of nonresponsiveness may have occurred extrathymically: the early neonatal thymus (presumably the source of the peripheral T cells observed in neoTx mice) also contains T cells with self-reactive receptors, but these cells are fully functional. Therefore, neonatal thymectomy aborts deletion of self-reactive T cells, but self-tolerance is maintained through functional inactivation of potentially self-reactive clones.

Human airway smooth muscle is structurally and mechanically similar to that of other species.

Airway smooth muscle (ASM) plays a vital role in the exaggerated airway narrowing seen in asthma. However, whether asthmatic ASM is mechanically different from nonasthmatic ASM is unclear. Much of our current understanding about ASM mechanics comes from measurements made in other species. Limited data on human ASM mechanics prevents proper comparisons between healthy and asthmatic tissues, as well as human and animal tissues. In the current study, we sought to define the mechanical properties of healthy human ASM using tissue from intact lungs and compare these properties to measurements in other species. The mechanical properties measured included: maximal stress generation, force-length properties, the ability of the muscle to undergo length adaptation, the ability of the muscle to recover from an oscillatory strain, shortening velocity and maximal shortening. The ultrastructure of the cells was also examined. Healthy human ASM was found to be mechanically and ultrastructurally similar to that of other species. It is capable of undergoing length adaptation and responds to mechanical perturbation like ASM from other species. Force generation, shortening capacity and velocity were all similar to other mammalian ASM. These results suggest that human ASM shares similar contractile mechanisms with other animal species and provides an important dataset for comparisons with animal models of disease and asthmatic ASM.

Microfluidic droplet grating for reconfigurable optical diffraction.

This Letter presents a reconfigurable optical diffraction grating using multiphase droplets on a microfluidic chip. The uniform and evenly spaced circular droplets are generated by continuously dispersing two immiscible liquids into a T junction to produce plugs, which are then transformed into a circular shape at a sudden expansion of the microchannel. In experiments, the droplet grating shows a detection limit of ~6.3x10(-5) when used as an opto fl uidic refractometer and produces different colors as a color filter. Such a grating has the advantages of high stability and wide tunability in droplet size, grating period, and refractive index, making it promising for biochemical and biomaterial applications.

Use and abuse of faecal occult blood tests in an acute hospital inpatient setting.

BACKGROUND/AIMS: The role of the faecal occult blood test (FOBT) is untested. The aims of this study were to define the use of FOBT in a general hospital setting and to determine its influence on patient management. METHODS: Case notes and laboratory reports were retrospectively reviewed in all FOBTs performed in 2006 across three acute hospitals, with specific reference to clinical setting, indication, influence over clinical decision-making and management. Both guaiac and immunological tests were performed on all specimens. RESULTS: A total of 330 patients aged 2-104 (mean 74) years, 47% men, had 461 tests performed. A positive result was recorded in one or both tests in 64% of patients. Evidence of dietary restriction was found in only eight (2%) of patients and 218 (66%) patients took one or more medications that could have caused a false positive result. Indications were mostly for overt or suspected gastrointestinal blood loss with or without anaemia and/or iron deficiency, but 5% were for non-bloody diarrhoea and 3% screening for colorectal cancer. Patient care was adversely affected or delayed in 54 patients (16%), mostly because of the result being the stimulus for the decision to refer or not for endoscopy. Only one was considered appropriate as a screening test for colorectal cancer. CONCLUSIONS: The FOBT was applied in clinically inappropriate settings without consideration to confounding issues, and often led to inappropriate clinical decisions with considerable cost to hospital and patient. There is no place for FOBT in an acute hospital setting.

Cardiopulmonary exercise test indices of respiratory buffering before and after aerobic exercise training in women with pulmonary hypertension: Differentiation by magnitudes of change in six-minute walk test performance.

While aerobic exercise training (AET) has generally been shown to improve 6-min walk test (6MWT) distance (6MWD) in patients with pulmonary hypertension (PH), a substantial number of patients appear to adapt differently, with minimal or even negative changes in 6MWT distance being reported. PURPOSE: To compare post-aerobic exercise training adaptations in cardiorespiratory functional capacity across three groups of patients with PH: those with high (HI), low (LI) and negative (NEG) post-training increases in 6MWD. METHODS: Participants were 25 females (age 54 ± 11 years; BMI 31 ± 7 kg/m2) who completed a vigorous, 10-week, thrice weekly, supervised treadmill walking exercise program. Cardiopulmonary exercise tests (CPET) and 6MWT were completed before and after training. Ten of the 25 participants were classified as HI (range = 47-143 m), 11 were classified as LI (range = 4-37 m) and 4 were classified as NEG (range = -17 to -53 m). RESULTS: Peak CPET duration, WR and time to anaerobic threshold (AT) were significantly higher (p < 0.05) after training in both the LI and HI groups but not in the NEG group. There was a significant improvement in VE/VCO2 (p = 0.042), PETCO2 (p = 0.011) and TV (p = 0.050) in the HI group after training, but not in the NEG or LI group. CONCLUSION: These findings suggest that sustained ventilatory inefficiency and restricted respiratory buffering may mediate exercise intolerance and impede the ability to adapt to exercise training in some patients with PH.

Synaptic vesicle recycling in synapsin I knock-out mice.

The synapsins are a family of four neuron-specific phosphoproteins that have been implicated in the regulation of neurotransmitter release. Nevertheless, knock-out mice lacking synapsin Ia and Ib, family members that are major substrates for cAMP and Ca2+/ Calmodulin (CaM)-dependent protein kinases, show limited phenotypic changes when analyzed electrophysiologically (Rosahl, T.W., D. Spillane, M. Missler, J. Herz, D.K. Selig, J.R. Wolff, R.E. Hammer, R.C. Malenka, and T.C. Sudhof. 1995. Nature (Lond.). 375: 488-493; Rosahl, T.W., M. Geppert, D. Spillane, D., J. Herz, R.E. Hammer, R.C. Malenka, and T.C. Sudhof. 1993. Cell. 75:661-670; Li, L., L.S. Chin, O. Shupliakov, L. Brodin, T.S. Sihra, O. Hvalby, V. Jensen, D. Zheng, J.O. McNamara, P. Greengard, and P. Andersen. 1995. Proc. Natl. Acad. Sci. USA. 92:9235-9239; see also Pieribone, V.A., O. Shupliakov, L. Brodin, S. Hilfiker-Rothenfluh, A.J. Czernik, and P. Greengard. 1995. Nature (Lond.). 375:493-497). Here, using the optical tracer FM 1-43, we characterize the details of synaptic vesicle recycling at individual synaptic boutons in hippocampal cell cultures derived from mice lacking synapsin I or wild-type equivalents. These studies show that both the number of vesicles exocytosed during brief action potential trains and the total recycling vesicle pool are significantly reduced in the synapsin I-deficient mice, while the kinetics of endocytosis and synaptic vesicle repriming appear normal.