When good organs go to bad people.
A number of philosophers have argued that alcoholics should receive lower priority for liver transplantations because they are morally responsible for their medical conditions. In this paper, I argue that this conclusion is false. Moral responsibility should not be used as a criterion for the allocation of medical resources. The reason I advance goes further than the technical problem of assessing moral responsibility. The deeper problem is that using moral responsibility as an allocation criterion undermines the functioning of medicine.


Functional expression and characterization of skeletal muscle dihydropyridine receptors in Xenopus oocytes.
Dihydropyridine receptors in vertebrate skeletal muscle serve a dual role: as voltage sensors for excitation-contraction coupling and as voltage-activated calcium channels. Although they were the first of six classes of calcium channels to be cloned, skeletal muscle dihydropyridine receptors remain the only ones not functionally expressed as calcium channels in Xenopus oocytes, leading to the hypothesis that an interacting component is missing. Using beta1b, an isoform previously found in brain, we have for the first time reconstituted skeletal muscle calcium channel function in Xenopus oocytes. We show that this beta subunit is necessary for functional expression and that the alpha2delta subunit significantly enhances the expressed current. The majority of the alpha1 subunit in skeletal muscle is a truncated form. Here we show that both the full-length and truncated forms produce functional calcium channels in Xenopus oocytes, but the truncated form gives significantly larger currents. In addition, we show that the beta1b transcript is expressed in rat skeletal muscle, although at a much lower level than the abundant beta1a isoform.


Factors related to the donor organ are major determinants of renal allograft function and survival.
In this study, we analyzed the relative impact of donor and recipient variables on cadaveric renal allograft function and survival. The unique feature of the study population is that each pair of recipients received their allografts from a single donor. The study includes 378 adult patients. In 129 pairs both recipients were Caucasian, and in 60 pairs one recipient was Caucasian and the other was African-American. All transplants were done in one center, thus minimizing differences in preservation time and providing uniform posttransplant management. The initial analysis showed a relationship between the function of the allograft 6 months after transplantation (serum creatinine [SCr]6 mo) and donor variables (P = 0.0004, analysis of variance). Furthermore, it was calculated that 64% of the variability in the SCr 6mo among patients was due to donor factors and 36% was due to recipient factors. An elevated SCr 6 mo was significantly associated with older donors, male recipients, and patients with acute rejection episodes. Furthermore, other unidentified donor factors may have an impact on allograft function. Reflecting the importance of donor factors, there was a significant relationship between SCr 6mo in paired recipients (P < 0.0008 by Spearman). Analysis of racially dissimilar pairs showed that the SCr 6mo and graft survival 6 months after transplantation were not significantly different between Caucasians and African-Americans. However, beyond 6 months, graft survival was worse in African-Americans (P < 0.0001 by Cox). Compared with Caucasians, graft survival was significantly worse in African-Americans with poorly controlled blood pressure (mean arterial pressure > 105 mmHg) (P = 0.002, Cox), but not in those patients with mean arterial pressure < 105 mmHg. In conclusion, donor factors are major determinants of renal allograft function. However, those factors may not be easily identifiable or quantifiable. Donor factors do not contribute to racial differences in allograft survival. However, poorly controlled hypertension correlates with poor renal graft survival in African-Americans.


[Possible role of embryonic surface antigens in forming the autostimulus of tumor cell proliferation].
It is shown that DNA synthesis intesifies in a mixed culture of cells of the tumours of different histogenesis, induced by monkey adenovirus SA7(C8) and chemical carcinogens, with syngenic and allogenic cells of an early embryo. This intensification of the synthesis in tumour cells is due to the contact of surfaces participating in the cell response but not to the soluble factors because when the embryo cells are placed into the milliporous chamber, the described effect is inhibited.


EARLY FLOWERING 3 and Photoperiod Sensing in Brachypodium distachyon.
The proper timing of flowering, which is key to maximize reproductive success and yield, relies in many plant species on the coordination between environmental cues and endogenous developmental programs. The perception of changes in day length is one of the most reliable cues of seasonal change, and this involves the interplay between the sensing of light signals and the circadian clock. Here, we describe a Brachypodium distachyon mutant allele of the evening complex protein EARLY FLOWERING 3 (ELF3). We show that the elf3 mutant flowers more rapidly than wild type plants in short days as well as under longer photoperiods but, in very long (20 h) days, flowering is equally rapid in elf3 and wild type. Furthermore, flowering in the elf3 mutant is still sensitive to vernalization, but not to ambient temperature changes. Molecular analyses revealed that the expression of a short-day marker gene is suppressed in elf3 grown in short days, and the expression patterns of clock genes and flowering time regulators are altered. We also explored the mechanisms of photoperiodic perception in temperate grasses by exposing B. distachyon plants grown under a 12 h photoperiod to a daily night break consisting of a mixture of red and far-red light. We showed that 2 h breaks are sufficient to accelerate flowering in B. distachyon under non-inductive photoperiods and that this acceleration of flowering is mediated by red light. Finally, we discuss advances and perspectives for research on the perception of photoperiod in temperate grasses.


Blinded comparison of a direct immunofluorescent monoclonal antibody staining method and a Giemsa staining method for identification of Pneumocystis carinii in induced sputum and bronchoalveolar lavage specimens of patients infected with human immunodeficiency virus.
A new direct immunofluorescence monoclonal antibody (DFA) method (Genetic Systems, Inc., Seattle, Wash.) for identification of Pneumocystis carinii in induced sputum and bronchoalveolar lavage specimens was compared in a blinded study with an established Giemsa stain method. We evaluated 148 consecutive clinical specimens from 104 patients with the following results. For the 67 patients (64%) infected with the human immunodeficiency virus (HIV), 49 were initially negative by both the DFA and the Giemsa methods, none were negative by DFA and positive by Giemsa, 6 were positive by DFA and negative by Giemsa, and 12 were positive by both methods, for a sensitivity and a negative predictive value of greater than 99%. For the six patients positive by DFA and negative by Giemsa, all were positive by both methods on evaluation of subsequently obtained clinical specimens, suggesting a specificity of greater than 99% and a false-positive rate of less than 1%. For 37 patients whose HIV status was negative or unknown, 35 were negative by both methods and 2 were positive by DFA and negative by Giemsa. The DFA method was simple to perform and required less time for scoring of stained slides than the Giemsa method, but care had to be taken to avoid false-positive readings due to extraneous fluorescence. This study indicates that the DFA method represents an advance as a sensitive, simple, and rapid way to identify P. carinii in induced sputum and bronchoalveolar lavage specimens from HIV-infected patients and suggests greater sensitivity of the DFA than the Giemsa method in this patient population.


Efficacy and safety of Ascoril expectorant and other cough formula in the treatment of cough management in paediatric and adult patients--a randomised double-blind comparative trial.
The present study was undertaken to study the comparative safety and efficacy of two cough formulas viz, Ascoril expectorant and other cough formula in the management of cough associated with respiratory disorders. Fifty patients having cough associated with various respiratory disorders like bronchitis and upper or lower respiratory tract infections were randomly divided into 2 equal groups and were treated with one of the two cough formulas viz, Ascoril cough formula and other cough formula in double-blind manner over a period of 15 days. The evaluation of improvement was carried out by a rating scale using three clinical parameters--cough, sputum and breathlessness. The physicians were asked to rate the effectiveness of the therapy and patients were asked to rate the acceptability of therapy using pre-defined operational criteria. It was observed that the improvement and symptom relief was almost immediate, quicker and better in the group receiving Ascoril as compared to other group. On effectiveness parameter, 96% of the physicians rated Ascoril as having either 'very high effectiveness or high effectiveness' as opposed to only 34% of the physicians who rated other cough formula as having 'high' or 'very high effectiveness'. While on parameter of acceptability, 96% of the patients rated acceptability of Ascoril as 'high' or 'good' as opposed to only 24% of the patients who rated other cough formula 'high' or 'good'. The findings of this study suggests that Ascoril cough formula has better efficacy as well as better patient acceptability. Thus, Ascoril cough formula is superior to other cough formula in management of cough associated with respiratory disorders.


Enhanced Antifouling Properties of Carbohydrate Coated Poly(ether sulfone) Membranes.
Poly(ether sulfone) membranes (PES) were modified with biologically active monosaccharides and disaccharides using aryldiazonium chemistry as a mild, one-step, surface-modification strategy. We previously proposed the modification of carbon, metals, and alloys with monosaccharides using the same method; herein, we demonstrate modification of PES membranes and the effect of chemisorbed carbohydrate layers on their resistance to biofouling. Glycosylated PES surfaces were characterized using spectroscopic methods and tested against their ability to interact with specific carbohydrate-binding proteins. Galactose-, mannose-, and lactose-modified PES surfaces were exposed to Bovine Serum Albumin (BSA) solutions to assess unspecific protein adsorption in the laboratory and were found to adsorb significantly lower amounts of BSA compared to bare membranes. The ability of molecular carbohydrate layers to impart antifouling properties was further tested in the field via long-term immersive tests at a wastewater treatment plant. A combination of ATP content assays, infrared spectroscopic characterization and He-ion microscopy (HIM) imaging were used to investigate biomass accumulation at membranes. We show that, beyond laboratory applications and in the case of complex aqueous environments that are rich in biomass such as wastewater effluent, we observe significantly lower biofouling at carbohydrate-modified PES than at bare PES membrane surfaces.


Low-frequency ultrasonic velocity measurements in human calcaneal trabecular bone.
Ultrasonic velocities were measured in three orthogonal directions for 17 cubes (2 x 2 x 2 cm approximately) of defatted calcaneal trabecular bone using a novel pulse transmission method with 37 kHz transducers. Since the wavelength was greater than the cross-sectional dimensions of the specimens, it was assumed that bar wave propagation was occurring and this allowed Young's modulus to be derived from velocity and apparent density. Velocity varied from 1585 +/- 104 m s-1 in axis 1 (proximo-distal) to 947 +/- 131 m s-1 in axis 3 (medio-lateral). Thus, the velocities measured in axis 3 were considerably lower than those typically seen in clinical measurements of the calcaneus. The derived Young's moduli ranged from 834 +/- 248 MPa (axis 1) to 299 +/- 98 MPa (axis 3), and were comparable in magnitude to some previously published data from mechanical testing. The results suggest that velocities measured with this technique do indeed correspond to bar velocities, and consequently that low-frequency ultrasound can be used to directly predict the mechanical properties of trabecular bone specimens in vitro. On the other hand, given the marked differences in geometry, wavelengths used and the presence of fat, it raises questions about the validity of applying the bar wave equation in the context of higher-frequency velocity measurements in the intact calcaneus in vivo, despite the fact that this has been found to be useful by a number of previous workers. This may require a review of the clinical implications of high-frequency ultrasound data.


Role of P2-purinergic receptors in rat Leydig cell steroidogenesis.
The present study investigated the effects of extracellular ATP on the intracellular calcium ion concentration ([Ca2+]i) and testosterone production in isolated adult rat Leydig cells. This nucleotide caused an increase in [Ca2+]i, with a maximal effect at a concentration of 100 microM ATP, comprising a rapid initial spike followed by a long-lasting plateau. The first rapid spike was dependent on the release of Ca2+ from internal stores, since it also occurred in Ca(2+)-free medium and was abolished after depletion of internal stores with thapsigargin. The second, long-lasting, phase was dependent on the influx of Ca2+ from the extracellular medium. After 3 h of incubation, extracellular ATP stimulated testosterone secretion in a dose-dependent manner, with a maximal effect at 100 microM. Activation of steroidogenesis by ATP was fully dependent on the presence of Ca2+ in the external medium. Among different nucleotides, only ATP, adenosine 5'-[gamma-thio]triphosphate, UTP, benzoylbenzoic-ATP and 2-methylthio-ATP were effective in inducing both the rise in [Ca2+]i and testosterone secretion. These effects were blocked by preincubation of Leydig cells with oxidized ATP, an inhibitor of the P2Z-purinergic receptor subtype. These results show that rat Leydig cells possess P2-purinergic receptors whose activation triggers an increase in [Ca2+]i due to the release of Ca2+ from internal stores and Ca2+ influx from the external medium. The stimulatory effect of extracellular ATP on testosterone secretion seems to be coupled to the influx of Ca2+ from the external medium.


Spatiotemporal patterns in the energy release of great earthquakes.
For the past 80 years, the energy released in great strike-slip and thrust earthquakes has occurred in alternating cycles of 20 to 30 years. This pattern suggests that a global transfer mechanism from poloidal to toroidal components of tectonic plate motions is operating on time scales of several decades. The increase in seismic activity in California in recent years may be related to an acceleration of global strike-slip moment release, as regions of shear deformation mature after being reached by stresses that have propagated away from regions of great subduction decoupling earthquakes in the 1960s.


The value of SPECT/CT imaging of lacrimal glands as a means of assessing the activity of Graves' orbitopathy.
Graves' orbitopathy (GO) is an autoimmune disease with mechanical impairment of orbital muscles and lacrimal gland dysfunction. The frequently used methods of assessing GO activity include Clinical Activity Score (CAS), CT, and MRI. These approaches are mainly associated with orbital muscles; however, there are not many studies that focus on the lacrimal gland inflammation of GO patients. The aim of this study is to assess the usefulness of 99mTc-DTPA single-photon emission (SPE) CT/CT in evaluating the lacrimal gland inflammation in GO, as compared with other methods. A retrospective analysis of 48 patients with active GO compared with 33 controls was conducted. All subjects underwent clinical-endocrinological analyses, CAS evaluation, CT scans, and SPECT/CT examination. Lacrimal gland dimensions were determined and analyzed. The lacrimal glands in patients with GO were significantly larger in all measured dimensions (P < 0.001) on CT scans relative to those in controls. Increased lacrimal gland diethylene triamine pentaacetic acid (DTPA) uptake ratios (P < 0.001) were displayed in active GO patients compared to controls and were also correlated with thyrotropin receptor antibody levels. The cut-off value for discriminating active and inactive disease was calculated to be 1.735, with specificity of 82.6% and sensitivity of 74.2%. SPECT/CT uptake ratios and CAS values were positively correlated in all GO patients. SPECT/CT uptake ratios were also positively correlated with CT measurements including lacrimal gland volume and coronal width in GO patients. These data indicated that lacrimal gland SPECT/CT images can serve as a good tool for assessing the inflammation and disease activity of GO.


Psycholinguistic Screening for Cognitive Decline in Cancer Survivors: A Feasibility Study.
To test the feasibility of using psycholinguistic speech analysis as a proxy for cognitive function in men undergoing treatment for prostate cancer. Audio-recorded speech samples were collected from 13 men enrolled in a parent study at the University of Kansas Cancer Center in Kansas City. Audio-recorded speech samples, collected from clinical interviews and in response to a prompt question during the parent study at two time points, were evaluated to determine feasibility relationships between neurocognitive and psycholinguistic measures. Correlations between neurocognitive and psycholinguistic measures were identified for prompted speech, but the strength of relationships varied between time points. No relationships were identified in clinical interview speech samples. Feasibility was demonstrated for recording, transcribing, and analyzing speech from clinical interviews, and results suggest relationships between neurocognitive and psycholinguistic measures in prompted speech. If validated, psycholinguistic assessments may be used to assess cognitive function in cancer survivors. Advances in natural language processing may provide opportunities for automated speech analyses for cancer treatment-related cognitive decline.


The effectiveness of basic life support training on nursing students' knowledge and basic life support practices: a non-randomized quasi-experimental study.
Provision of up-to-date information and skills training related to basic life support practices is very important for nursing students' professional development and practitioner and education related roles. The purpose of the study was to examine the effectiveness of basic life support training on knowledge and practices among nursing students. A non-randomized quasi-experimental design (One group pre-test-post-test) was used in this study. The study was conducted in the laboratory of an undergraduate nursing school. The sample consisted of a convenience sample of 1st-year students enrolled in the undergraduate nursing class. The study sample consisted of 65 nursing students. Basic life support training included both theoretical and practical components. The students' knowledge and practices were assessed before basic life support training. Data were collected using the knowledge assessment questionnaire. The practical skills for basic life support were observed and assessed using a checklist. The pre- and post-assessment practice scores were compared. After basic life support training, level of knowledge and practical skill scores were higher compared to pre-training scores (t= -12.442, p=0.000; t= -22.899, p=0.000). There was a significant and moderate association between the adult basic life support Knowledge Form scores and the adult basic life support practice assessment form scores obtained after the training (r = 0.39, p<0.01). The study showed that basic life support training improved knowledge and skills related to basic life support practices in nursing students. Periodic basic life support training is very important for competency in this area among nursing students.


[Effect of electroacupuncture intervention on ERK 1/2 signaling and TNF-α and IL-1β protein levels in the substantia Nigra in rats with Parkinson's Disease].
To observe the effect of electroacupuncture(EA) on phosphalized extracellular regulated protein kinases(p-ERK) 1/2 signaling pathway and tumor necrosis factor (TNF)-α and IL-1β protein levels in the substantia nigra (SN) in rats with Parkinson's disease (PD), so as to explore its mechanism underlying improvement of PD. 32 male SD rats were randomly and averagely divided into normal control group, sham-operation group, model group and EA group. PD model was established by intra-dermal-injection of rotenone(1 mg/kg, dissolved in DMSO and saline, concentration: 0. 25 mg/mL) at the nape, once daily for 14 days. EA (2 Hz, 2 mA) was applied to "Fengfu"(GV 16)and "Taichong"(LR 3) for 20 min, once daily for 14 days. For sham-operation group, subcutaneous injection of the same dose of DMSO and saline was given in the same way. The expression levels of tyrosine hydroxylase (TH), p-ERK 1/2, inflammatory cytokines TNF-α and IL-1P proteins of the SN tissue were detected using Western blot. The rat's horizontal and vertical movement ability was assessed using open-field tests. The horizontal and vertical movement scores were significantly lower in the model group than in the normal control group, and markedly higher in the EA group than in the model group (P<0. 05). Compared with the normal control group, the expression level of TH protein in the SN was significantly reduced in the model group( P<0. 05 ),while the expression levels of p-ERK 1/2, TNF-α and IL-1β proteins were significantly increased in the model group(P<0. 05). After EA intervention, the expression level of TH protein was significantly increased(P<0.05),and those of p-ERK 1/2, TNF-α and IL-1β proteins were significantly down-regulated(P<0. 05) in the EA group compared to the model group. EA therapy may improve PD rats' movement ability, which may be associated with its effects in down-regulating the expression levels of p-ERK 1/2, TNF-α and IL-1β proteins and up-regulating the expression of TH protein in the SN.


Evaluation of a social marketing intervention promoting oral rehydration salts in Burundi.
Diarrhea is the second leading cause of death for children under five in Burundi; however, use of oral rehydration salts (ORS), the recommended first-line treatment, remains low. In 2004, PSI/Burundi launched a social marketing intervention to promote ORASEL among caregivers of children under five; the product was relaunched in 2006 with a new flavor. This study evaluates the intervention after the ORASEL relaunch, which included mass media and interpersonal communication activities. The study looks at trends in ORASEL use in Burundi and in behavioral determinants that may be related to its use. In 2006 and 2007, PSI conducted household surveys among Burundian females of reproductive age (15-49). Both surveys used a two-stage sampling process to select 30 households in each of 115 rural and urban collines throughout the nation. Survey respondents were asked about diarrhea treatment-related behavior; key behavioral determinants; and exposure to the ORASEL intervention. Data were analyzed to identify trends over time, characteristics of ORASEL users, and associations between exposure to the intervention and changes in ORASEL use and related behavioral determinants. ORASEL use among caregivers at their children's last diarrheal episode increased significantly from 20% in 2006 to 30% in 2007, and there were also desirable changes in several behavioral determinants associated with ORASEL use. Evaluation analysis showed that a higher level of exposure to the social marketing campaign was associated with greater use of ORASEL and with significant improvements in perceived availability, knowledge of the signs of diarrhea and dehydration, social support, and self-efficacy. ORS use can be improved through social marketing and educational campaigns that make the public aware of the availability of the product, encourage dialogue about its use, and increase skills and confidence relating to correct product preparation and administration. Further interventions in Burundi and elsewhere should promote ORS through a variety of mass media and interpersonal communication channels, and should be rigorously evaluated in the context of the total market for diarrhea treatment products.


Biochemical mechanisms of cyclosporine neurotoxicity.
Proper management of chemotoxicity in transplant patients requires detailed knowledge of the biochemical mechanisms underlying immunosuppressant toxicity. Neurotoxicity is one of the most significant clinical side effects of the immunosuppressive undecapeptide cyclosporine, occurring at some degree in up to 60% of transplant patients. The clinical symptoms of cyclosporine-mediated neurotoxicity consist of decreased responsiveness, hallucinations, delusions, seizures, cortical blindness, and stroke-like episodes that mimic those clinical symptoms of mitochondrial encephalopathy. Clinical computed tomography (CT) and magnetic resonance imaging (MRI) studies have revealed a correlation between clinical symptoms of cyclosporine-mediated neurotoxicity and morphological changes in the brain, such as hypodensity of white matter, cerebral edema, metabolic encephalopathy, and hypoxic damages. Paradoxically, in animal models cyclosporine protects the brain from ischemia-reperfusion (I/R) injury. Interestingly, cyclosporine appears to mediate both neurotoxicity (under normoxic conditions) and I/R protection across the same range of drug concentration. Both toxicity and protection might arise from the intersection of cyclosporine with mitochondrial energy metabolism. This review addresses basic biochemical mechanisms of: 1) cyclosporine toxicity in normoxic brain, and 2) its protective effects in the same organ during I/R. The marked and unparallel potential of magnetic resonance spectroscopy (MRS) as a novel quantitative approach to evaluate metabolic drug toxicity is described.


Intracellular recognition of lipopolysaccharide by toll-like receptor 4 in intestinal epithelial cells.
Toll-like receptor (TLR)4 has recently been shown to reside in the Golgi apparatus of intestinal crypt epithelial m-ICcl2 cells, colocalizing with internalized lipopolysaccharide (LPS). Here we demonstrate that disruption of the integrity of the Golgi apparatus significantly reduced LPS-mediated nuclear factor kappaB activation. Also, the TLR4 adaptor protein MyD88 and the serine/threonine kinase IRAK-1 were rapidly recruited to the Golgi apparatus upon stimulation. LPS-mediated activation required lipid raft formation and intact clathrin-dependent internalization. In contrast to macrophages, prevention of ligand internalization by use of LPS-coated beads significantly impaired recognition by epithelial cells. The localization of TLR4 to the Golgi apparatus was abrogated by expression of a genetically modified form of the TLR4 binding chaperone gp96. Thus, our data provide evidence that in contrast to the situation in macrophages, LPS recognition in intestinal epithelial cells may occur in the Golgi apparatus and require LPS internalization.


A Soft Wearable Microfluidic Patch with Finger-Actuated Pumps and Valves for On-Demand, Longitudinal, and Multianalyte Sweat Sensing.
Easy sample collection, physiological relevance, and ability to noninvasively and longitudinally monitor the human body are some of the key attributes of wearable sweat sensors. Examples typically include reversible sensors or an array of single-use sensors embedded in specialized microfluidics for temporal analysis of sweat. However, evolving this field to a level that truly represents "lab-on-skin" technology will require the incorporation of advanced functionalities that give the user the freedom to (1) choose the precise time for performing sample analysis and (2) select sensors from an array embedded within the device for performing condition-specific sample analysis. Here, we introduce new concepts in wearable microfluidic platforms that offer such capabilities. The described technology involves a series of finger-actuated pumps, valves, and sensors incorporated within soft, wearable microfluidics. The incoming sweat collects in the inlet chamber and can be analyzed by the user at the time of their choosing. On-demand sweat analyte assessment is achieved by pulling a thin tab to activate a pump which opens a valve and allows the pooled sweat to enter a chamber embedded with sensors for the desired analytes. The article describes a thorough characterization of the platform that demonstrates the robustness of the pumping, valving, and sensing aspects of the device under conditions mimicking real-life scenarios. A two-day-long human pilot study validates the system and illustrates the device's ability to offer on-demand, longitudinal, and multianalyte sensing. Our work represents the first example of a wearable system with such on-demand sensing capabilities and opens exciting avenues in sweat sensing for acquiring new insights into human physiology.


Real-world medication use and economic outcomes in incident systemic lupus erythematosus patients in the United States.
Aims: Systemic lupus erythematosus (SLE) is a multifaceted autoimmune disease involving multiple organs systems and places a significant economic burden on SLE patients. There is a literature gap regarding the standard of care and economic burden in SLE patients, their families, and society. This study assessed medication use patterns among SLE patients and generated the annual and total economic burden associated with the illness.Materials and methods: Adult patients with ≥2 medical claims on different dates for SLE diagnoses were identified from 01 January 2013 to 31 December 2015 using two large administrative claims databases representative of the commercially insured US population. Patient demographics and clinical characteristics during 1-year pre-SLE diagnosis were assessed. Outcomes including the proportion of patients who used SLE medications and annual costs were assessed 1-year post-SLE diagnosis. Total costs related to SLE were extrapolated to the US population to estimate the economic burden based on SLE prevalence.Results: A total of 30,086 SLE patients were identified. The most common baseline comorbidities were hypertension and infections. Corticosteroids and hydroxychloroquine were the most common SLE medications. Biologics utilization was minimal. SLE patients had, on average, 26.0 physician visits, 23.7 prescription claims, 1.7 inpatient admissions, and 2.0 hospital days per patient 1-year post-SLE diagnosis. Annual all-cause median costs among all SLE patients were $8712 per patient per year. Total costs ranged between $1.4-1.6 and $2.8-3.2 billion per year, depending on prevalence estimates.Conclusions: Our findings indicate a nominal use of biologics (∼2%) among SLE patients; despite belimumab being one of the few approved treatments for SLE in the USA. These data reveal an unmet need for availability of advanced SLE therapy, and future studies are warranted concerning the underlying causes. SLE is also associated with a substantial economic burden of ≤3.2 billion per year. These findings may assist in future planning and resource allocation.


Ca(2+) uptake in the sarcoplasmic reticulum from the systemic heart of octopod cephalopods.
We have measured Ca(2+) uptake in crude homogenates of heart tissue, as well as cell shortening and ionic currents in isolated myocytes exposed to caffeine, to characterize Ca(2+) uptake in the sarcoplasmic reticulum (SR) of the systemic heart of octopus. The maximal rate of SR Ca(2+) uptake in crude homogenates of octopus heart was 43+/-4 (mean +/- s.e.m., N=7), compared with 28+/-2 nmol min(-)(1 )mg(-)(1) protein (N=4) in homogenates of rat heart. The Ca(2+)-dependency of SR Ca(2+) uptake was similar for the two species, with a Ca(2+) activity at half-maximal uptake rate (pCa(50)) of 6.04+/-0.02 for octopus and 6.02+/-0.05 for rat. Exposure of isolated myocytes to 10 mmol l(-)(1) caffeine resulted in cell shortening to 53+/-2 % of the resting cell length and an inward trans-sarcolemmal ionic current. The charge carried by this current was 3.28+/-0.70 pC pF(-)(1) (mean +/- s.e.m., N=5) corresponding to extrusion of 34.0+/-0.7 amol Ca(2+ )pF(-)(1) from the cell by Na(+)/Ca(2+) exchange. This is approximately 50 times more than the Ca(2+) carried by the Ca(2+) current elicited by a 200 ms depolarization from -80 to 0 mV and corresponds to an increase in the total intracellular [Ca(2+)] of 404+/-86 (&mgr;)mol l(-)(1) non-mitochondrial volume due to Ca(2+) release from the SR. Thus, we find that at 20 degrees C in the SR both Ca(2+) content and Ca(2+) uptake rate in the systemic heart of octopus are comparable with or larger than the corresponding values obtained in the rat heart. These results support the argument that the SR may play an important role in the regulation of contraction in the systemic heart of cephalopods.


Mental impact of Covid-19 among Spanish healthcare workers. A large longitudinal survey.
Longitudinal data on the mental health impact of the coronavirus disease 2019 (Covid-19) pandemic in healthcare workers is limited. We estimated prevalence, incidence and persistence of probable mental disorders in a cohort of Spanish healthcare workers (Covid-19 waves 1 and 2) -and identified associated risk factors. 8996 healthcare workers evaluated on 5 May-7 September 2020 (baseline) were invited to a second web-based survey (October-December 2020). Major depressive disorder (PHQ-8 ≥ 10), generalised anxiety disorder (GAD-7 ≥ 10), panic attacks, post-traumatic stress disorder (PCL-5 ≥ 7), and alcohol use disorder (CAGE-AID ≥ 2) were assessed. Distal (pre-pandemic) and proximal (pandemic) risk factors were included. We estimated the incidence of probable mental disorders (among those without disorders at baseline) and persistence (among those with disorders at baseline). Logistic regression of individual-level [odds ratios (OR)] and population-level (population attributable risk proportions) associations were estimated, adjusting by all distal risk factors, health care centre and time of baseline interview. 4809 healthcare workers participated at four months follow-up (cooperation rate = 65.7%; mean = 120 days s.d. = 22 days from baseline assessment). Follow-up prevalence of any disorder was 41.5%, (v. 45.4% at baseline, p < 0.001); incidence, 19.7% (s.e. = 1.6) and persistence, 67.7% (s.e. = 2.3). Proximal factors showing significant bivariate-adjusted associations with incidence included: work-related factors [prioritising Covid-19 patients (OR = 1.62)], stress factors [personal health-related stress (OR = 1.61)], interpersonal stress (OR = 1.53) and financial factors [significant income loss (OR = 1.37)]. Risk factors associated with persistence were largely similar. Our study indicates that the prevalence of probable mental disorders among Spanish healthcare workers during the second wave of the Covid-19 pandemic was similarly high to that after the first wave. This was in good part due to the persistence of mental disorders detected at the baseline, but with a relevant incidence of about 1 in 5 of HCWs without mental disorders during the first wave of the Covid-19 pandemic. Health-related factors, work-related factors and interpersonal stress are important risks of persistence of mental disorders and of incidence of mental disorders. Adequately addressing these factors might have prevented a considerable amount of mental health impact of the pandemic among this vulnerable population. Addressing health-related stress, work-related factors and interpersonal stress might reduce the prevalence of these disorders substantially. Study registration number: NCT04556565.


Epigenetics of reproductive infertility.
Infertility is a complex pathophysiological condition. It may caused by specific or multiple physical and physiological factors, including abnormalities in homeostasis, hormonal imbalances and genetic alterations. In recent times various studies implicated that, aberrant epigenetic mechanisms are associated with reproductive infertility. There might be transgenerational effects associated with epigenetic modifications of gametes and studies suggest the importance of alterations in epigenetic modification at early and late stages of gametogenesis. To determine the causes of infertility it is necessary to understand the altered epigenetic modifications of associated gene and mechanisms involved therein. This review is devoted to elucidate the recent mechanistic advances in regulation of genes by epigenetic modification and emphasizes their possible role related to reproductive infertility. It includes environmental, nutritional, hormonal and physiological factors and influence of internal structural architecture of chromatin nucleosomes affecting DNA and histone modifications in both male and female gametes, early embryogenesis and offspring. Finally, we would like to emphasize that research on human infertility by gene knock out of epigenetic modifiers genes must be relied upon animal models.


Effects of apomorphine and haloperidol on the acoustic startle response in rats.
A series of 3 experiments tested the effects of 0.01, 0.04, 0.19, 0.75, 3.00, and 6.00 mg/kg apomorphine and 0.13, 0.25, and 0.50 haloperidol on the acoustic startle response in rats. Apomorphine markedly facilitated startle amplitude for about 40 min after injection and then depressed startle over the next 40 min. Both the early facilitory and later inhibitory effects were directly related to the dose. Haloperidol (0.5 mg/kg--given 30 min before) completely blocked both the early facilitory and the later depressant effect of apomorphine (3 mg/kg). Haloperidol alone had only a slight depressant effect on startle. The data support the conclusion that DA receptor stimulation enhances acoustic startle amplitude and indicate that a previous report failed to find an effect of apomorphine on startle because startle was only measured 40 min after injection.


Identification of clinical features and autoantibodies associated with calcinosis in dermatomyositis.
Prior studies have estimated that up to 20% of adults with dermatomyositis (DM) have calcinosis, which can lead to significant morbidity. Identification of risk factors may provide a better understanding of the pathogenesis and ultimately therapy for this difficult clinical problem. Risk factors for calcinosis in adults with DM have not been extensively studied. To determine the prevalence of calcinosis and to identify associated clinical features in a cohort of extensively phenotyped adults with DM. A cross-sectional study of 126 patients diagnosed as having DM from January 1, 2006, through January 1, 2013, was performed. Patients were adults (≥18 years of age) attending the Stanford University Medical Center clinic. Calcinosis, defined as the presence of calcium deposition in the skin and subcutaneous tissues on physical examination. Fourteen patients (11.1%) had calcinosis, with the extremities most commonly involved. Patients with vs those without calcinosis had a longer disease duration (median, 6.9 years; range, 2.4-18.1; vs median, 3.9 years; range, 0.2-19.2 years; P = .003) and more fingertip ulcers (50.0% vs 9.3%, P < .001). An association between calcinosis and both interstitial lung disease and anti-MDA-5 autoantibodies was identified, but this association did not persist in multivariate models that adjusted for fingertip ulcers. Fingertip ulcers and disease duration were strongly associated with calcinosis in all multivariate models, independent of the underlying autoantibody present. Autoantibodies to NXP-2 were associated with calcinosis (odds ratio, 15.52; 95% CI, 2.01-119.90), whereas anti-transcriptional intermediary factor 1-γ antibodies were protective (odds ratio, 0.2; 95% CI, 0.01-0.99) in multivariate analyses that adjusted for fingertip ulcers and other covariates. Calcinosis was a relatively uncommon clinical feature in our cohort of adults with DM. Our data suggest that calcinosis is positively associated with longer disease duration, fingertip ulcers, and NXP-2 autoantibodies and negatively associated with transcriptional intermediary factor 1-γ antibodies. A common vascular mechanism may underlie the development of both calcinosis and fingertip ulcers in patients with DM.


Functional connectivity of parietal cortex during temporal selective attention.
Perception of natural experiences requires allocation of attention towards features, objects, and events that are moving and changing over time. This allocation of attention is controlled by large-scale brain networks that, when damaged, cause widespread cognitive deficits. In particular, damage to ventral parietal cortex (right lateralized TPJ, STS, supramarginal and angular gyri) is associated with failures to selectively attend to and isolate features embedded within rapidly changing visual sequences (Battelli, Pascual-Leone, & Cavanagh, 2007; Husain, Shapiro, Martin, & Kennard, 1997). In this study, we used fMRI to investigate the neural activity and functional connectivity of intact parietal cortex while typical subjects judged the relative onsets and offsets of rapidly flickering tokens (a phase discrimination task in which right parietal patients are impaired). We found two regions in parietal cortex correlated with task performance: a bilateral posterior TPJ (pTPJ) and an anterior right-lateralized TPJ (R aTPJ). Both regions were deactivated when subjects engaged in the task but showed different patterns of functional connectivity. The bilateral pTPJ was strongly connected to nodes within the default mode network (DMN) and the R aTPJ was connected to the attention network. Accurate phase discriminations were associated with increased functional correlations between sensory cortex (hMT+) and the bilateral pTPJ, whereas accuracy on a control task was associated with yoked activity in the hMT+ and the R aTPJ. We conclude that temporal selective attention is particularly sensitive for revealing information pathways between sensory and core cognitive control networks that, when damaged, can lead to nonspatial attention impairments in right parietal stroke patients.


Reducing racism in Aboriginal health care in Australia: where does cultural education fit?
This paper discusses whether educating health professionals and undergraduate students in culturally respectful health service delivery is effective in reducing racism, improving practice and lessening the disparities in health care between Aboriginal and non-Aboriginal Australians. The paper supports the concept of race as a social construction that is discursively produced and reproduced. Studies on the effectiveness of cross-cultural education for undergraduate students and health professionals to reduce racism and deliver culturally respectful health care to indigenous or minority populations are examined for evidence of sustained improvements to practice. Programs in culturally respectful health care delivery can lead to short-term improvements to practice. Sustained change is more elusive as few programs conducted long-term evaluations. Long-term evaluation of programs in culturally respectful health care delivery is necessary to identify whether early changes to behavior and practices are sustained. Strategies linking policies to practice to reduce health disparities between Aboriginal and non-Aboriginal Australians are also needed. Confronting the effects of racism in health services towards Aboriginal Australians is a priority requiring a multi-tiered commitment to strategies linking policy to practice to reduce health disparities between Aboriginal and non-Aboriginal Australians. Part of this strategy includes preparing undergraduates and health professionals for culturally respectful health care with education programs that are evaluated for long-term improvements to practice.


[Histo-topography of the lymphoid tissue of children's trachea].
By means of certain microscopical methods areas from the middle third of trachea have been examined in children, died from various causes not connected with diseases in the pulmonary pathways. At birth in the mucous membrane of the trachea there is no lymphoid tissue. Its elements in a small amount appear at the initial stage of the suckling period. In two-month-old infants the first signs of lymphoid perinucleoli appear. During the early period of infancy the main regions of the lymphoid cells localization are clearly defined, in the area of some ducts of the tracheal glands including. In children, whose age corresponds to the first period of childhood, lymphoid accumulations have a pattern, resembling similar structures of the trachea in mature persons and the greatest concentration of cells, in comparison to the preceding groups of children in the ages examined.


Statin use and decreased risk of benign prostatic enlargement and lower urinary tract symptoms.
To determine whether statin use is associated with a decreased risk of developing benign prostatic enlargement (BPE) and lower urinary tract symptoms (LUTS). We conducted a retrospective, population-based cohort study of 2447 men, 40-79 years of age, residing in Olmsted County, MN, USA, in 1990, and followed these men biennially through 2007. Cox proportional hazard models were used to assess associations between statin use and new onset of moderate/severe LUTS (American Urological Association Symptom Index score >7), a decreased maximum urinary flow rate (<12 mL/s) or BPE (prostate volume >30 mL). Statin use was inversely associated with new onset of LUTS (Hazard ratio (HR) 0.39; 95% confidence interval (CI) 0.31-0.49), a decreased maximum flow rate (HR 0.53; 95% CI 0.34-0.82) and BPE (HR 0.40; 95% CI 0.23-0.69) after adjustment for baseline age and body mass index, diabetes, hypertension, coronary heart disease, smoking, alcohol use, activity level and non-steroidal anti-inflammatory use. The longest duration of statin use was associated with the lowest risk of developing each outcome (all tests for trend: P < 0.001). In this study, statin use was associated with a 6.5- to 7-year delay in the new onset of moderate/severe LUTS or BPE. While men typically take statin medications to prevent coronary heart disease events and related outcomes, these data suggest that men who use statins may also receive urologic benefits.


Potential reverse spillover of infectious bursal disease virus at the interface of commercial poultry and wild birds.
Recently, multiple spillover events between domesticated poultry and wild birds have been reported for several avian viruses. This phenomenon highlights the importance of the livestock-wildlife interface in the possible emergence of novel viruses. The aim of the current study was to investigate the potential spillover and epidemiological links of infectious bursal disease virus (IBDV) between wild birds and domestic poultry. To this end, twenty-eight cloacal swabs were collected from four species of free-living Egyptian wild birds (i.e. mallard duck, bean goose, white-fronted goose and black-billed magpie). Genetic and phylogenetic analysis of three positive isolates revealed that the IBDV/USC-1/2019 strain clustered with previously reported very virulent IBDV (vvIBDV) Egyptian isolates. Interestingly, two other wild bird-origin isolates (i.e. IBDV/USC-2/2019 and IBDV/USC-3/2019) grouped with a vaccine strain that is being used in commercial poultry. In conclusion, our results revealed the molecular detection of vaccine and vvIBDV-like strains in Egyptian wild birds and highlighted the potential role of wild birds in IBDV epidemiology in disease-endemic regions.


Method for Vertical Calibration of Laser-Projection Transnasal Fiberoptic High-Speed Videoendoscopy.
The ability to provide absolute calibrated measurement of the laryngeal structures during phonation is of paramount importance to voice science and clinical practice. Calibrated three-dimensional measurement could provide essential information for modeling purposes, for studying the developmental aspects of vocal fold vibration, for refining functional voice assessment and treatment outcomes evaluation, and for more accurate staging and grading of laryngeal disease. Recently, a laser-calibrated transnasal fiberoptic endoscope compatible with high-speed videoendoscopy (HSV) and capable of providing three-dimensional measurements was developed. The optical principle employed is to project a grid of 7 × 7 green laser points across the field of view (FOV) at an angle relative to the imaging axis, such that (after calibration) the position of each laser point within the FOV encodes the vertical distance from the tip of the endoscope to the laryngeal tissues. The purpose of this study was to develop a precise method for vertical calibration of the endoscope. Investigating the position of the laser points showed that, besides the vertical distance, they also depend on the parameters of the lens coupler, including the FOV position within the image frame and the rotation angle of the endoscope. The presented automatic calibration method was developed to compensate for the effect of these parameters. Statistical image processing and pattern recognition were used to detect the FOV, the center of FOV, and the fiducial marker. This step normalizes the HSV frames to a standard coordinate system and removes the dependence of the laser-point positions on the parameters of the lens coupler. Then, using a statistical learning technique, a calibration protocol was developed to model the trajectories of all laser points as the working distance was varied. Finally, a set of experiments was conducted to measure the accuracy and reliability of every step of the procedure. The system was able to measure absolute vertical distance with mean percent error in the range of 1.7% to 4.7%, depending on the working distance.


A machine learning-based prediction of the micropapillary/solid growth pattern in invasive lung adenocarcinoma with radiomics.
Micropapillary/solid (MP/S) growth patterns of lung adenocarcinoma are vital for making clinical decisions regarding surgical intervention. This study aimed to predict the presence of a MP/S component in lung adenocarcinoma using radiomics analysis. Between January 2011 and December 2013, patients undergoing curative invasive lung adenocarcinoma resection were included. Using the "PyRadiomics" package, we extracted 90 radiomics features from the preoperative computed tomography (CT) images. Subsequently, four prediction models were built by utilizing conventional machine learning approaches fitting into radiomics analysis: a generalized linear model (GLM), Naïve Bayes, support vector machine (SVM), and random forest classifiers. The models' accuracy was assessed using a receiver operating curve (ROC) analysis, and the models' stability was validated both internally and externally. A total of 268 patients were included as a primary cohort, and 36.6% (98/268) of them had lung adenocarcinoma with an MP/S component. Patients with an MP/S component had a higher rate of lymph node metastasis (18.4% versus 5.3%) and worse recurrence-free and overall survival. Five radiomics features were selected for model building, and in the internal validation, the four models achieved comparable performance of MP/S prediction in terms of area under the curve (AUC): GLM, 0.74 [95% confidence interval (CI): 0.65-0.83]; Naïve Bayes, 0.75 (95% CI: 0.65-0.85); SVM, 0.73 (95% CI: 0.61-0.83); and random forest, 0.72 (95% CI: 0.63-0.81). External validation was performed using a test cohort with 193 patients, and the AUC values were 0.70, 0.72, 0.73, and 0.69 for Naïve Bayes, SVM, random forest, and GLM, respectively. Radiomics-based machine learning approach is a very strong tool for preoperatively predicting the presence of MP/S growth patterns in lung adenocarcinoma, and can help customize treatment and surveillance strategies.


Simultaneous bio-reduction of trichloroethene, trichloroethane, and chloroform using a hydrogen-based membrane biofilm reactor.
The contamination of water by chlorinated solvents is recognized as a serious and widespread problem throughout the industrialized world. Here, we focus on three chlorinated solvents that are among those most commonly detected and that have distinct chemical features: trichloroethene (TCE), trichloroethane (TCA), and chloroform (CF). Because many contaminated waters contain mixtures of the chlorinated solvents, a treatment technology that detoxifies all of them simultaneously is highly desirable. The membrane biofilm reactor (MBfR) is a recent technological advance that makes it possible to deliver H(2) gas to bacteria efficiently and safely, despite hydrogen's low water solubility and risk of forming a combustible atmosphere when mixed with air. The objectives of this work are to document whether or not the three chlorinated compounds can be dechlorinated simultaneously in a H(2)-based MBfR and to determine if competitive or inhibitory interactions affect bio-reduction of any of the solvents. The main finding is a demonstration that directly using H(2) as the electron donor makes it possible to bio-reduce combinations of different chlorinated solvents. This finding supports that the H(2)-based MBfR can treat multiple chlorinated solvents in one step, addressing a common groundwater situation. We saw possible evidence of inhibition by CF at a concentration greater than about 1 muM, competition for H(2) from sulfate and nitrate reductions, and possible inhibition of TCE reduction from the accumulation of chloroethane (CA) or chloromethane (CM).


[Fractures of the atlantoaxial complex in the elderly: assessment of radiological spectrum of fractures and factors influencing imaging diagnosis].
To describe patterns of atlantoaxial fractures in a population of consecutive elderly patients, including assessment of type, distribution and associated clinical and radiological findings, and to analyze any influence of the causative trauma mechanism on the individual fracture pattern. The distribution and type of 123 atlantoaxial fractures in 95 subjects older than 65 years (range: 65 - 102; mean age: 79 years) were retrospectively assessed. For each subject, trauma mechanism and clinical and neurological status were recorded at admission. Initial imaging studies of the cervical spine were reviewed. Preexistent degenerative changes were assessed and the atlantoaxial fractures classified. Data were evaluated for the frequency of different types of fractures of C1 and C2 and for accompanying fractures of cervical vertebrae or the occipital condyles, respectively. The majority of patients with injuries of the atlantoaxial complex had fractures of C2 (90 of 95, 95 %). A large proportion of these patients (67 of 90, 74 %) had odontoid fractures. An isolated fracture of C1 was present in only 5 (5 %) patients. Associated fractures of the occipital condyles or other cervical vertebrae were rare (10 of 95, 11 %). The main trauma mechanism for atlantoaxial injuries was a fall (56 of 95, 59 %). Elderly patients injured in motor vehicle accidents were more likely to have isolated fractures of C2 and Type III fractures of the odontoid (p < 0.02). In elderly patients, fractures of the atlantoaxial complex are mainly caused by falls and almost always involve C2. The trauma mechanism influences the fracture pattern.


The effect of meal frequency on biochemical cardiometabolic factors: A systematic review and meta-analysis of randomized controlled trials.
Although several randomized controlled trials (RCTs) have supported the beneficial effects of higher meal frequency (MF) on cardiometabolic risk factors, the putative effects of higher MF on health remain inconclusive. This study systematically reviewed the evidence from RCTs of the effect of higher compared with lower MF on the blood lipid profile, glucose homeostasis, and adipokines. PubMed, Scopus, ISI Web of Science, and the Cochrane database were searched up to October 2020 to retrieve relevant RCTs. A DerSimonian and Laird random effects model was used to pool mean differences and 95% CI for each outcome. The quality of studies and evidence was assessed through standard methods. Twenty-one RCTs (686 participants) were included in this meta-analysis. Overall results showed a significant improvement in total cholesterol [weighted mean difference (WMD) = -6.08 mg/dl; 95% CI: -10.68, -1.48; P = 0.01; I2 = 88%], and low-density cholesterol (LDL-C) (WMD = -6.82 mg/dl; 95% CI: -10.97, -1.60; P = 0.009; I2 = 85.7%), while LDL-C to high-density cholesterol ratio (LDL-C: HDL-C) increased (WMD = 0.22; 95% CI: 0.07, 0.36; P = 0.003; I2 = 0.0%) in higher MF vs. lower MF. No significant effects were found on measures of glycemic control, apolipoproteins-A1 and B, or leptin. In subgroup analyses, higher MF significantly reduced serum triglyceride (TG), and increased HDL-C, compared with lower MF in interventions > 12 weeks, and decreased serum TC and LDL-C in healthy participants. A significant reduction in LDL-C also was observed in studies where the same foods given both arms, simply divided into different feeding occasions, and in feeding studies, following higher MF compared to lower MF. Our meta-analysis found that higher, compared with lower MF may improve total cholesterol, and LDL-C. The intervention does not affect measures of glycemic control, apolipoproteins-A1 and B, or leptin. However, the GRADE ratings of low credibility of the currently available evidence highlights the need for more high-quality studies in order to reach a firm conclusion.


Should remaining stockpiles of smallpox virus (variola) be destroyed?
In 2011, the World Health Organization will recommend the fate of existing smallpox stockpiles, but circumstances have changed since the complete destruction of these cultures was first proposed. Recent studies suggest that variola and its experimental surrogate, vaccinia, have a remarkable ability to modify the human immune response through complex mechanisms that scientists are only just beginning to unravel. Further study that might require intact virus is essential. Moreover, modern science now has the capability to recreate smallpox or a smallpox-like organism in the laboratory in addition to the risk of nature re-creating it as it did once before. These factors strongly suggest that relegating smallpox to the autoclave of extinction would be ill advised.


A system to simultaneously detect tick-borne pathogens based on the variability of the 16S ribosomal genes.
DNA microarrays can be used to quickly and sensitively identify several different pathogens in one step. Our previously developed DNA microarray, based on the detection of variable regions in the 16S rDNA gene (rrs), which are specific for each selected bacterial genus, allowed the concurrent detection of Borrelia spp., Anaplasma spp., Francisella spp., Rickettsia spp. and Coxiella spp. In this study, we developed a comprehensive detection system consisting of a second generation DNA microarray and quantitative PCRs. New oligonucleotide capture probes specific for Borrelia burgdorferi s.l. genospecies and Candidatus Neoehrlichia mikurensis were included. This new DNA microarray system required substantial changes in solution composition, hybridization conditions and post-hybridization washes. This second generation chip displayed high specificity and sensitivity. The specificity of the capture probes was tested by hybridizing the DNA microarrays with Cy5-labeled, PCR-generated amplicons encoding the rrs genes of both target and non-target bacteria. The detection limit was determined to be 10(3) genome copies, which corresponds to 1-2 pg of DNA. A given sample was evaluated as positive if its mean fluorescence was at least 10% of the mean fluorescence of a positive control. Those samples with fluorescence close to the threshold were further analyzed using quantitative PCRs, developed to identify Francisella spp., Rickettsia spp. and Coxiella spp. Like the DNA microarray, the qPCRs were based on the genus specific variable regions of the rrs gene. No unspecific cross-reactions were detected. The detection limit for Francisella spp. was determined to be only 1 genome copy, for Coxiella spp. 10 copies, and for Rickettsia spp., 100 copies. Our detection system offers a rapid method for the comprehensive identification of tick-borne bacteria, which is applicable to clinical samples. It can also be used to identify both pathogenic and endosymbiontic bacteria in ticks for eco-epidemiological studies, tick laboratory colony testing, and many other applications.


Growth hormone deficiency, secondary hypothyroidism, and empty sella following treatment of childhood macroprolactinoma.
Macroprolactinoma are rare in childhood, especially in the first decade. A 9-year-old girl presented with headache, vomiting, and decreased vision for 8 months. A diagnosis of macroprolactinoma was made following documentation of elevated serum prolactin (958 ng/ml) with a contrast enhancing macroadenoma (30 × 27 × 28 mm) on magnetic resonance imaging of pituitary. Anterior pituitary function was normal. Cabergoline therapy resulted in resolution of all symptoms in 2-8 months. Revaluation at 10 months of cabergoline therapy revealed normal serum prolactin (14 ng/ml), normal pituitary function, with 91% decrease in adenoma size (11.5 × 13.6 × 12.7 mm). Evaluation at 36 months of cabergoline therapy for growth arrest and weight gain for past 6 months revealed low serum prolactin, growth hormone deficiency, and secondary hypothyroidism with empty sella. She had biochemical as well as structural resolution of prolactinoma. This report highlights the development of multiple pituitary hormone deficiency with empty sella, an uncommon side effect of cabergoline therapy for macroprolactinoma.


Non-equilibrium theory of the allele frequency spectrum.
A forward diffusion equation describing the evolution of the allele frequency spectrum is presented. The influx of mutations is accounted for by imposing a suitable boundary condition. For a Wright-Fisher diffusion with or without selection and varying population size, the boundary condition is lim(x downward arrow0)xf(x,t)=thetarho(t), where f(.,t) is the frequency spectrum of derived alleles at independent loci at time t and rho(t) is the relative population size at time t. When population size and selection intensity are independent of time, the forward equation is equivalent to the backwards diffusion usually used to derive the frequency spectrum, but this approach allows computation of the time dependence of the spectrum both before an equilibrium is attained and when population size and selection intensity vary with time. From the diffusion equation, a set of ordinary differential equations for the moments of f(.,t) is derived and the expected spectrum of a finite sample is expressed in terms of those moments. The use of the forward equation is illustrated by considering neutral and selected alleles in a highly simplified model of human history. For example, it is shown that approximately 30% of the expected total heterozygosity of neutral loci is attributable to mutations that arose since the onset of population growth in roughly the last 150,000 years.


Measurement of internal tissue optical properties at ultraviolet and visible wavelengths: Development and implementation of a fiberoptic-based system.
A novel, multi-wavelength, fiberoptic system was constructed, evaluated and implemented to determine internal tissue optical properties at ultraviolet A (UVA) and visible (VIS) wavelengths. Inverse modeling was performed with a neural network to estimate absorption and reduced scattering coefficients based on spatially-resolved reflectance distributions. The model was calibrated with simulated reflectance datasets generated using a condensed Monte Carlo approach with absorption coefficients up to 85 cm(-1) and reduced scattering coefficients up to 118 cm(-1). After theoretical and experimental evaluations of the system, optical properties of porcine bladder, colon, esophagus, oral mucosa, and liver were measured at 325, 375, 405, 445 and 532 nm. These data provide evidence that as wavelengths decrease into the UVA, the dominant tissue chromophore shifts from hemoglobin to structural proteins such as collagen. This system provides a high level of accuracy over a wide range of optical properties, and should be particularly useful for in situ characterization of highly attenuating biological tissues in the UVA-VIS.


A meta-analysis of the vascular-related safety profile and efficacy of alpha-adrenergic blockers for symptoms related to benign prostatic hyperplasia.
To evaluate the safety profile and efficacy of alpha1-adrenergic receptor blockers (A1Bs) currently prescribed for benign prostatic hyperplasia (BPH). A systematic literature search of MEDLINE, the Cochrane Database and the Food and Drug Administration Web site through December 2006 identified double-blinded, prospective, placebo-controlled trials, evaluating agents commercially available by prescription for the symptomatic treatment of BPH. Data were reviewed by two investigators with the use of a standardised data abstraction form. Studies were evaluated for methodological quality using the Jadad scale. Studies with a score of < 3 were considered of weaker methodology. Of 2389 potential citations, 25 were usable for evaluation of safety data, 26 for efficacy. A1B use was associated with a statistically significant increase in the odds of developing a vascular-related event [odds ratio (OR) 2.54; 95% confidence interval (CI): 2.00-3.24; p < 0.0001]. The odds of developing a vascular-related adverse event were: alfuzosin, OR 1.66, 95% CI: 1.17-2.36; terazosin, OR 3.71, 95% CI: 2.48-5.53; doxazosin, OR 3.32, 95% CI: 2.10-5.23 and tamsulosin, OR 1.42, 95% CI: 0.99-2.05. A1Bs increased Q(max) by 1.32 ml/min (95% CI: 1.07-1.57) compared with placebo. Difference from placebo in American Urological Association symptom index/International Prostate Symptom Score was -1.92 points (95% CI: -2.71 to -1.14). Alfuzosin, terazosin and doxazosin showed a statistically significant increased risk of developing vascular-related events compared with placebo. Tamsulosin showed a numerical increase that was not statistically significant. All agents significantly improved Q(max) and symptom signs compared with placebo.


Dietary enrichment with fish oil prevents high fat-induced metabolic dysfunction in skeletal muscle in mice.
High saturated fat (HF-S) diets increase intramyocellular lipid, an effect ameliorated by omega-3 fatty acids in vitro and in vivo, though little is known about sex- and muscle fiber type-specific effects. We compared effects of standard chow, HF-S, and 7.5% HF-S replaced with fish oil (HF-FO) diets on the metabolic profile and lipid metabolism gene and protein content in red (soleus) and white (extensor digitorum longus) muscles of male and female C57BL/6 mice (n = 9-12/group). Weight gain was similar in HF-S- and HF-FO-fed groups. HF-S feeding increased mesenteric fat mass and lipid marker, Oil Red O, in red and mixed muscle; HF-FO increased interscapular brown fat mass. Compared to chow, HF-S and HF-FO increased expression of genes regulating triacylglycerol synthesis and fatty acid transport, HF-S suppressed genes and proteins regulating fatty acid oxidation, whereas HF-FO increased oxidative genes, proteins and enzymes and lipolytic gene content, whilst suppressing lipogenic genes. In comparison to HF-S, HF-FO further increased fat transporters, markers of fatty acid oxidation and mitochondrial content, and reduced lipogenic genes. No diet-by-sex interactions were observed. Neither diet influenced fiber type composition. However, some interactions between muscle type and diet were observed. HF-S induced changes in triacylglycerol synthesis and lipogenic genes in red, but not white, muscle, and mitochondrial biogenesis and oxidative genes were suppressed by HF-S and increased by HF-FO in red muscle only. In conclusion, HF-S feeding promotes lipid storage in red muscle, an effect abrogated by the fish oil, which increases mediators of lipolysis, oxidation and thermogenesis while inhibiting lipogenic genes. Greater storage and synthesis, and lower oxidative genes in red, but not white, muscle likely contribute to lipid accretion encountered in red muscle. Despite several gender-dimorphic genes, both sexes exhibited a similar HF-S-induced metabolic and gene expression profile; likewise fish oil was similarly protective in both sexes.


Fabrication and Characterization of Novel Poly(D-lactic acid) Nanocomposite Membrane for Water Filtration Purpose.
The development of membrane technology from biopolymer for water filtration has received a great deal of attention from researchers and scientists, owing to the growing awareness of environmental protection. The present investigation is aimed at producing poly(D-lactic acid) (PDLA) membranes, incorporated with nanocrystalline cellulose (NCC) and cellulose nanowhisker (CNW) at different loadings of 1 wt.% (PDNC-I, PDNW-I) and 2 wt.% (PDNC-II PDNW-II). From morphological characterization, it was evident that the nanocellulose particles induced pore formation within structure of the membrane. Furthermore, the greater surface reactivity of CNW particles facilitates in enhancing the surface wettability of membranes due to increased hydrophilicity. In addition, both thermal and mechanical properties for all nanocellulose filled membranes under investigation demonstrated significant improvement, particularly for PDNW-I-based membranes, which showed improvement in both aspects. The membrane of PDNW-I presented water permeability of 41.92 L/m2h, when applied under a pressure range of 0.1-0.5 MPa. The investigation clearly demonstrates that CNWs-filled PDLA membranes fabricated for this investigation have a very high potential to be utilized for water filtration purpose in the future.


Changes in the blood and plasma volume of Harris' sparrows during postnuptial molt.
In captive Harris' sparrows (Zonotrichia querula), plasma and blood volume increased significantly during stages 2 and 3 of postnuptial molt, when extensive feather replacement was taking place in the wings, tail and body tracts. Erythrocyte number did not increase significantly until molt was essentially complete. Consequently the 6% mean maximum reduction in hematocrit that was observed during molt appeared to be largely the result of plasma volume expansion. This plasma volume expansion was not due to changes in the osmolality or concentration of proteins within the blood plasma since both decreased significantly during intense molt.


Androgen receptor expression is inversely correlated with pathologic tumor stage in bladder cancer.
To evaluate the expression of the androgen receptor (AR) in transitional cell carcinoma (TCC) of the bladder, and to assess whether its expression correlated with pathologic tumor stage. TCC of the bladder is three times more common in males than in females. The origin of this sex difference in incidence is unknown. We evaluated tumor specimens from 49 consecutive patients treated for TCC of the bladder at our institution between July 2002 and June 2003. Immunohistochemistry was performed using a monoclonal mouse anti-AR antibody on paraffin-embedded tissue sections of tumors obtained from transurethral resection, radical cystectomy, or resection of metastases. Specimens were assessed for AR expression, and, in tumors that demonstrated AR staining, the percentage of nuclei that stained positive was recorded. Of the 49 tumors, 26 (53.1%) expressed the AR. The percentage of tumors that expressed the AR decreased with increasing pathologic stage, from 88.9% of pTa lesions to 0% of pT3 tumors. Overall, 75% of superficial tumors (pTa + pT1 + carcinoma in situ) expressed the AR compared with 21.4% of invasive tumors (pT2 + pT3; P = 0.002). In addition, among AR-expressing tumors, the mean percentage of nuclei that stained positive for the AR was significantly greater in pTa tumors (62.5%) than in pT1 (31%) or pT2 (20%) tumors (P = 0.005). We found a decrease in AR protein expression in tumors with increased pathologic stage. Our data suggest that the loss of AR expression is associated with invasive bladder cancer.


Epitope Mapping of PMab-225 an Anti-Alpaca Podoplanin Monoclonal Antibody Using Flow Cytometry.
Podoplanin (PDPN) is a mucin-type membrane glycoprotein, and possesses three platelet aggregation-stimulating (PLAG) domains: PLAG1, PLAG2, and PLAG3 at the N-terminus of PDPN, and one or two PLAG-like domains (PLDs) in the middle of PDPN. PDPN is expressed on normal tissues, such as podocytes of the kidney and type I alveolar cells of the lung, and is also overexpressed in numerous malignant cancers. Previously, we reported a novel anti-alpaca podoplanin (aPDPN) monoclonal antibody (mAb), PMab-225, using Cell-Based Immunization and Screening (CBIS) method. PMab-225 specifically detected aPDPN-overexpressed Chinese hamster ovary (CHO)-K1 (CHO/aPDPN) cells using flow cytometry and western blotting, and strongly stained alpaca tissues such as lung type I alveolar cells by immunohistochemistry. However, the specific binding epitope of aPDPN for PMab-225 remains unclear. Thus, in this study, a series of deletion or point mutations of aPDPN were utilized for investigating the binding epitope of PMab-225 using flow cytometry. The analysis of deletion mutants showed that N-terminus of PMab-225 epitope might exist between 80 amino acid (aa) and 85 aa of aPDPN. Furthermore, the analysis of point mutants demonstrated that Thr84 of aPDPN, which exists in PLD, could be included in the critical epitope of PMab-225.


Correlations between cancer mortality rates from various Italian regions.
Death certification rates from 17 non-sexual and 4 sexual cancers were used to examine patterns of correlation between various cancers within the 20 Italian regions. A large number of strongly positive correlations emerged, reflecting the geographical distribution of cancer mortality in Italy which shows substantially higher rates for several common sites in northern areas. The most notable findings were the high positive correlations between various tobacco-related cancers in both sexes (however somewhat higher in males), the positive correlations between most intestinal sites and between a well defined group of other cancers including intestines in both sexes, breast and ovary in females and prostate in males, previously described in several widely heterogeneous populations. Various alcohol-related cancers showed high positive coefficients in males but not in females. Several suggestions which emerged from previous correlation studies but which generally lacked convincing biological or epidemiological consistency were not confirmed by the present data. Conversely, a few strong correlations emerged in the present study which are not explainable in terms of available knowledge of the causes of cancer, or obvious confounding. Though probably incidental, the existence of these correlations between cancers with widely heterogeneous or largely undefined etiology is still an indirect indication that these neoplasms are largely avoidable, since it is unlikely that the same genetic determinants are strongly associated with such different malignancies.


Manipulating flow separation: sensitivity of stagnation points, separatrix angles and recirculation area to steady actuation.
A variational technique is used to derive analytical expressions for the sensitivity of several geometric indicators of flow separation to steady actuation. Considering the boundary layer flow above a wall-mounted bump, the six following representative quantities are considered: the locations of the separation point and reattachment point connected by the separatrix, the separation angles at these stagnation points, the backflow area and the recirculation area. For each geometric quantity, linear sensitivity analysis allows us to identify regions which are the most sensitive to volume forcing and wall blowing/suction. Validations against full nonlinear Navier-Stokes calculations show excellent agreement for small-amplitude control for all considered indicators. With very resemblant sensitivity maps, the reattachment point, the backflow and recirculation areas are seen to be easily manipulated. By contrast, the upstream separation point and the separatrix angles are seen to remain extremely robust with respect to external steady actuation.


Visualization of cardiac ventricular myosin heavy chain homodimers and heterodimers by monoclonal antibody epitope mapping.
Two mAbs, one specific for cardiac alpha-myosin heavy chains (MHC) and the other specific for cardiac beta-MHC, were used to investigate the heavy-chain dimeric organization of rat cardiac ventricular myosin. Epitopes of the two mAbs were mapped on the myosin molecule by electron microscopy of rotary shadowed mAb-myosin complexes. mAbs were clearly identifiable by the different locations of their binding sites on the myosin rod. Thus, myosin molecules could be directly discriminated according to their alpha-or beta-MHC content. alpha alpha-MHC and beta beta-MHC homodimers were visualized in complexes consisting of two molecules of the same mAb bound to one myosin molecule. By simultaneously using the alpha-MHC-specific mAb and the beta-MHC-specific mAb, alpha beta-MHC heterodimers were visualized in complexes formed by one molecule of each of the two mAbs bound to one myosin molecule. Proportions of alpha alpha-and beta beta-MHC homodimers and alpha beta-MHC heterodimers were estimated from quantifications of mAb-myosin complexes and compared with the proportions given by electrophoreses under nondenaturing conditions. This visualization of cardiac myosin molecules clearly demonstrates the arrangement of alpha- and beta-MHC in alpha alpha-MHC homodimers, beta beta-MHC homodimers, and alpha beta-MHC heterodimers, as initially proposed by Hoh, J. F. Y., G. P. S. Yeoh, M. A. W. Thomas, and L. Higginbottom (1979).


mTOR: A Novel Therapeutic Target for Diseases of Multiple Systems.
Significant progress in the research of mammalian target of rapamycin (mTOR) in recent years, has greatly enhanced our understanding of the role and cellular pathways through which mTOR control cellular processes, such as translational initiation, actin organization, cell proliferation, and cell survival. mTOR is activated by phosphorylation and functions mainly through mTOR complex 1 or mTOR complex 2. mTORC1 is activated through tuberous sclerosis complex 1/2 dependent and independent mechanisms following the stimulation by growth factors, nutrient, amino acids, and other signaling pathways. The activity of mTOR is closely associated with cell proliferation and differentiation, apoptosis, and autophagy. Activation of mTOR prevents the induction of both apoptosis and autophagy through regulating its multiple targets. Given that the activity of mTOR has been involved in the pathogenesis of neurodegenerative disorders, cardiovascular abnormalities, metabolic diseases, renal transplantation, autoimmune abnormalities, and cancer, manipulating mTOR activation may represent as an innovative therapeutic strategy for these diseases. Yet, the role of mTOR in the body is complicated and therefore, its activity needs to be tightly regulated to achieve beneficial outcome in a specific pathological condition.


Hepatic disorders associated with liver-kidney microsomal antibodies.
A study of the clinical associations of a recently defined tissue autoantibody, the liver/kidney microsomal (L.K.M.) antibody, showed that out of 33 patients 26 had clinical liver disease. Fifteen of the patients had active chronic hepatitis and there were seven cases of acute hepatitis, precipitated by presumed virus A infection in three instances and by drug hypersensitivity in the other four. The remaining cases with liver disease included two with subclinical hepatitis and two with hepatocellular carcinoma. Evidence is presented that the patients with active chronic hepatitis may represent a distinct subgroup of the disease with a young mean age, an even male to female ratio, and a striking lack of other nonorgan-specific autoantibodies-that is, antinuclear and smooth muscle-which are usually present in the other autoimmune variant of the disease.


Supporting the moral development of medical students.
Philosophers who studied moral development have found that individuals normally progress rapidly in early adulthood from a conventional stage in which they base behavior on the norms and values of those around them to a more principled stage where they identify and attempt to live by personal moral values. Available data suggest that many medical students, who should be in this transition, show little change in their moral development. Possibly, this relates to perceived pressures to conform to the informal culture of the medical wards. Many students experience considerable internal dissidence as they struggle to accommodate personal values related to empathy, care, and compassion to their clinical training. Educational interventions that positively influence this process have established regular opportunities for critical reflection by the students in small groups. Other interventions include faculty development to enhance role modeling and feedback by clinical faculty. The author espouses more widespread adoption of these educational interventions.


Milk Powder Fortified with Potassium and Phytosterols to Decrease the Risk of Cardiovascular Events among the Adult Population in Malaysia: A Cost-Effectiveness Analysis.
This study evaluated the cost-effectiveness of the consumption of a milk powder product fortified with potassium (+1050.28 mg/day) and phytosterols (+1200 mg/day) to lower systolic blood pressure and low-density lipoprotein cholesterol, respectively, and, therefore, the risk of myocardial infarction (MI) and stroke among the 35-75-year-old population in Malaysia. A Markov model was created against a do-nothing option, from a governmental perspective, and with a time horizon of 40 years. Different data sources, encompassing clinical studies, practice guidelines, grey literature, and statistical yearbooks, were used. Sensitivity analyses were performed to evaluate the impact of uncertainty on the base case estimates. With an incremental cost-effectiveness ratio equal to international dollars (int$) 22,518.03 per quality-adjusted life-years gained, the intervention can be classified as very cost-effective. If adopted nationwide, it would help prevent at least 13,400 MIs, 30,500 strokes, and more than 10,600 and 17,100 MI- and stroke-related deaths. The discounted cost savings generated for the health care system by those who consume the fortified milk powder would amount to int$8.1 per person, corresponding to 0.7% of the total yearly health expenditure per capita. Sensitivity analyses confirmed the robustness of the results. Together with other preventive interventions, the consumption of milk powder fortified with potassium and phytosterols represents a cost-effective strategy to attenuate the rapid increase in cardiovascular burden in Malaysia.


Neuroendocrine Cells Are Commonly Absent in the Intestinal Crypts in Autoimmune Enteropathy.
The absence of neuroendocrine (NE) cells in the intestinal mucosa in autoimmune enteropathy (AIE) has been occasionally reported. However, the status of NE cells has not been studied in detail in AIE. Small bowel and colonic biopsies were retrospectively retrieved from 18 AIE patients (26 baseline [18 small bowel and 8 colon]; and 15 follow-up [11 duodenum and 4 colon] biopsies in 11 patients). Thirty-three common variable immunodeficiency (CVID) patients (30 small bowel and 16 colon), 15 inflammatory bowel disease patients (5 duodenum and 10 colon), 13 immunoglobulinA deficiency patients (13 duodenum and 5 colon), and 10 normal controls (5 colon and 5 duodenum) were selected as control groups. Histologic features (villous atrophy, intraepithelial lymphocytosis, acute inflammation, crypt apoptosis, and absence or presence of goblet cells, Paneth cells and plasma cells) were recorded. Chromogranin immunostain was performed and chromogranin-positive NE cells were counted per 10 consecutive, well-oriented crypts. On the basis of the number of chromogranin-positive NE cells, cases were graded as being absent (≤3 NE cells), markedly decreased (≤15), and intact (>15). The NE cell status correlated with histologic features. The median age of 18 AIE patients was 38.5 years (range: 11 to 74 y) and 14 patients were male. Fourteen of 18 (78%) patients showed loss (absent or markedly decreased) of NE cells in the small bowel and/or colon in the baseline biopsies including 12 (of 18) small bowel and 6 (of 8) colon biopsies. Follow-up biopsy was available in 11 patients. Six of 7 (85%) patients who showed loss of NE cells in the baseline biopsies regained NE cells in the follow-up biopsies, and 1 patient continued to show loss of NE cells. Four patients who showed intact NE cells in the baseline remained unchanged in the follow-up. Among the control groups, 3 of 33 (9%) CVID patients showed loss of NE cells. NE cells were not lost in the biopsies of all 15 and 13 patients with inflammatory bowel disease and immunoglobulinA deficiency, respectively, or the 10 normal controls. In all 41 biopsies (26 baseline plus 15 follow-up) with AIE, NE cell loss was significantly associated with increased crypt apoptosis and loss of goblet cells (P=0.001, both) but not with other histologic findings. In conclusion, our study suggests that NE cells may also be the target cells in AIE and commonly lost in the intestinal crypts in AIE, and consequently loss of NE cells can be used as an adjunct histologic feature for diagnosis of AIE.


Association of suboptimal prescribing and change in lower extremity physical function over time.
Previous studies have found inconsistent links between suboptimal prescribing and negative patient outcomes. While suboptimal prescribing consists of multiple components, e.g. drugs to avoid in the elderly (DAE), potential drug interactions (PDI) and polypharmacy, most research has focused on the impact of drugs to avoid. This study explores the relationship between suboptimal prescribing, comorbid disease, and change in lower extremity functional limitation (LEFL). This prospective cohort study used data from the Hispanic Established Population for the Epidemiologic Study of the Elderly. Baseline data collection occurred between 1993 and 1994 with three additional waves of data collected approximately every 2 years. Based on the disablement process model, the dependent variable was change in LEFL over the 7-year study period. Independent variables included suboptimal prescribing: DAE, PDI and polypharmacy. Measures of pathology included comorbid diseases (stroke, cancer, hypertension, cardiovascular disease, arthritis, and diabetes). Age, gender, education, smoking, cognitive status, depression, body mass index, marital status, and self-reported health were controlled in analyses. Diabetes, stroke, and arthritis were associated with a decline in LEFL. Polypharmacy mediated the relationship between diabetes and LEFL, and polypharmacy was also significantly associated with decrements in LEFL. The effect of suboptimal prescribing on change in LEFL was limited to both direct and mediational effects of polypharmacy. Additional research exploring the association between suboptimal prescribing and a variety of quality measures using a diverse set of outcomes would improve our understanding of the impact of suboptimal prescribing more broadly defined.


Issues of livelihood, sustainable development, and governance: Bay of Bengal.
This article is based on the findings of the Global International Waters Assessment (GIWA) Subregion 53, Bay of Bengal. It introduces the Subregion. The wide disparity in development indicators in the Bay of Bengal Subregion (BOBSR) is presented. The large population of poor people living in South Asia is presented as a factor that needs special attention. The article focuses on the 3 geographic sites selected for detailed analysis: i) the Ganges-Brahmaputra-Meghna river systems; ii) the Merbok Estuary mangroves, Malaysia; and iii) the Sunderbans mangroves, Bangladesh. Integrated water management based upon regional cooperation among Bangladesh, India and Nepal holds opportunities for mutual benefit. Policy options are proposed. For mangrove ecosystems, the impacts of urbanization in Malaysia and the unmanaged expansion of shrimp farming in Bangladesh are analyzed. Improved governance was seen to hold promise for enhancing economic benefits from shrimp farming while safeguarding the natural ecological system. However, these measures need to be a part of national efforts to achieve the UN Millennium Development Goals.


Outpatient cervical conization.
One hundred outpatient cervical conizations were performed over a 2.5-year period. A circumferential suturing technique was used which provided excellent hemostasis and restoration of normal cervical anatomy. The complication rate was low. One patient had significant intraoperative hemorrhage, one experienced postoperative bleeding within 24 hours, and two had delayed bleeding. Three patients developed infection, and one patient was found to have asymptomatic cervical stenosis on follow-up examination. No patient required blood transfusion or admission to the hospital. It is concluded that cervical conization can be performed on an outpatient basis in the nonpregnant patient with an acceptable rate of complications.


Cloning of a catabolite repression control (crc) gene from Pseudomonas aeruginosa, expression of the gene in Escherichia coli, and identification of the gene product in Pseudomonas aeruginosa.
Mutants which are defective in catabolite repression control (CRC) of multiple independently regulated catabolic pathways have been previously described. The mutations were mapped at 11 min on the Pseudomonas aeruginosa chromosome and designated crc. This report describes the cloning of a gene which restores normal CRC to these Crc- mutants in trans. The gene expressing this CRC activity was subcloned on a 2-kb piece of DNA. When this 2-kb fragment was placed in a plasmid behind a phage T7 promoter and transcribed by T7 RNA polymerase, a soluble protein with a molecular weight (MW) of about 30,000 was produced in Escherichia coli. A soluble protein of identical size was overproduced in a Crc- mutant when it contained the 2-kb fragment on a multicopy plasmid. This protein could not be detected in the mutant containing the vector without the 2-kb insert or with no plasmid. When a 0.3-kb AccI fragment was removed from the crc gene and replaced with a kanamycin resistance cassette, the interrupted crc gene no longer restored CRC to the mutant, and the mutant containing the interrupted gene no longer overproduced the 30,000-MW protein. Pools of intracellular cyclic AMP and the activities of adenylate cyclase and phosphodiesterase were measured in mutant and wild-type strains with and without a plasmid containing the crc gene. No consistent differences between any strains were found in any case. These results provide original evidence for a 30,000-MW protein encoded by crc+ that is required for wild-type CRC in P. aeruginosa and confirms earlier reports that the mode of CRC is cyclic AMP independent in this bacterium.


In vitro binding of Helicobacter pylori to human gastric mucin.
The in vitro binding of four Helicobacter pylori strains to human gastric mucin was studied with an enzyme-linked immunosorbent assay. All four strains were found to bind to purified mucin. Neuraminidase treatment and nonspecific oxidation of mucin decreased bacterial adherence to the macromolecule. Mucin preparations were also found to inhibit attachment of H. pylori to HEp-2 monolayers.


Genome analysis of the proteorhodopsin-containing marine bacterium Polaribacter sp. MED152 (Flavobacteria).
Analysis of marine cyanobacteria and proteobacteria genomes has provided a profound understanding of the life strategies of these organisms and their ecotype differentiation and metabolisms. However, a comparable analysis of the Bacteroidetes, the third major bacterioplankton group, is still lacking. In the present paper, we report on the genome of Polaribacter sp. strain MED152. On the one hand, MED152 contains a substantial number of genes for attachment to surfaces or particles, gliding motility, and polymer degradation. This agrees with the currently assumed life strategy of marine Bacteroidetes. On the other hand, it contains the proteorhodopsin gene, together with a remarkable suite of genes to sense and respond to light, which may provide a survival advantage in the nutrient-poor sun-lit ocean surface when in search of fresh particles to colonize. Furthermore, an increase in CO(2) fixation in the light suggests that the limited central metabolism is complemented by anaplerotic inorganic carbon fixation. This is mediated by a unique combination of membrane transporters and carboxylases. This suggests a dual life strategy that, if confirmed experimentally, would be notably different from what is known of the two other main bacterial groups (the autotrophic cyanobacteria and the heterotrophic proteobacteria) in the surface oceans. The Polaribacter genome provides insights into the physiological capabilities of proteorhodopsin-containing bacteria. The genome will serve as a model to study the cellular and molecular processes in bacteria that express proteorhodopsin, their adaptation to the oceanic environment, and their role in carbon-cycling.


Combinatorial QSAR modeling of specificity and subtype selectivity of ligands binding to serotonin receptors 5HT1E and 5HT1F.
The Quantitative Structure-Activity Relationship (QSAR) approach has been applied to model binding affinity and receptor subtype selectivity of human 5HT1E and 5HT1F receptor-ligands. The experimental data were obtained from the PDSP Ki Database. Several descriptor types and data-mining approaches have been used in the context of combinatorial QSAR modeling. Data mining approaches included k Nearest Neighbor, Automated Lazy Learning (ALL), and PLS; descriptor types included MolConnZ, MOE, DRAGON, Frequent Subgraphs (FSG), and Molecular Hologram Fingerprints (MHFs). Highly predictive QSAR models were generated for all three data sets (i.e., for ligands of both receptor subtypes and for subtype selectivity), and different individual techniques were proved best in each case. For real value activity data available for 5HT1E and 5HT1F ligand binding, models were characterized by leave-one-out cross-validated R(2) (q(2)) for the training sets and predictive R(2) values for the test sets. The best models for 5HT1E ligands were obtained with the kNN approach combined with MolConnZ descriptors (q(2)=0.69, R(2)=0.92); for 5HT1F ligands ALL QSAR method using MolConnZ descriptors gave the best results (R(2)=0.92). Rigorously validated classification models were also developed for the 5HT1E/5HT1F subtype selectivity data set with high correct classification accuracy for both training (CCRtrain=0.88) and test (CCRtest=1.00) sets using kNN with MolConnZ descriptors. The external predictive power of QSAR models was further validated by virtual screening of The Scripps Research Institute (TSRI) screening library to recover 5HT1E agonists and antagonists (not present in the original PDSP data set) with high enrichment factors. The successful development of externally predictive and interpretative QSAR models affords further design and discovery of novel subtype specific GPCR agents.


Age-based soldier polyethism: old termite soldiers take more risks than young soldiers.
Who should take on risky tasks in an age-heterogeneous society? Life-history theory predicts that, in social insects, riskier tasks should be undertaken by sterile individuals with a shorter life expectancy. The loss of individuals with shorter life expectancy is less costly for colony reproductive success than the loss of individuals with longer life expectancy. Termite colonies have a sterile soldier caste, specialized defenders engaged in the most risky tasks. Here we show that termite soldiers exhibit age-dependent polyethism, as old soldiers are engaged in front-line defence more than young soldiers. Our nest defence experiment showed that old soldiers went to the front line and blocked the nest opening against approaching predatory ants more often than young soldiers. We also found that young soldiers were more biased toward choosing central nest defence as royal guards than old soldiers. These results demonstrate that termite soldiers have age-based task allocation, by which ageing predisposes soldiers to switch to more dangerous tasks. This age-dependent soldier task allocation increases the life expectancy of soldiers, allowing them to promote their lifetime contribution to colony reproductive success.