Cellular and genetic diversity in the progression of in situ human breast carcinomas to an invasive phenotype.
Intratumor genetic heterogeneity is a key mechanism underlying tumor progression and therapeutic resistance. The prevailing model for explaining intratumor diversity, the clonal evolution model, has recently been challenged by proponents of the cancer stem cell hypothesis. To investigate this issue, we performed combined analyses of markers associated with cellular differentiation states and genotypic alterations in human breast carcinomas and evaluated diversity with ecological and evolutionary methods. Our analyses showed a high degree of genetic heterogeneity both within and between distinct tumor cell populations that were defined based on markers of cellular phenotypes including stem cell-like characteristics. In several tumors, stem cell-like and more-differentiated cancer cell populations were genetically distinct, leading us to question the validity of a simple differentiation hierarchy-based cancer stem cell model. The degree of diversity correlated with clinically relevant breast tumor subtypes and in some tumors was markedly different between the in situ and invasive cell populations. We also found that diversity measures were associated with clinical variables. Our findings highlight the importance of genetic diversity in intratumor heterogeneity and the value of analyzing tumors as distinct populations of cancer cells to more effectively plan treatments.


Autocorrelation descriptor improvements for QSAR: 2DA_Sign and 3DA_Sign.
Quantitative structure-activity relationship (QSAR) is a branch of computer aided drug discovery that relates chemical structures to biological activity. Two well established and related QSAR descriptors are two- and three-dimensional autocorrelation (2DA and 3DA). These descriptors encode the relative position of atoms or atom properties by calculating the separation between atom pairs in terms of number of bonds (2DA) or Euclidean distance (3DA). The sums of all values computed for a given small molecule are collected in a histogram. Atom properties can be added with a coefficient that is the product of atom properties for each pair. This procedure can lead to information loss when signed atom properties are considered such as partial charge. For example, the product of two positive charges is indistinguishable from the product of two equivalent negative charges. In this paper, we present variations of 2DA and 3DA called 2DA_Sign and 3DA_Sign that avoid information loss by splitting unique sign pairs into individual histograms. We evaluate these variations with models trained on nine datasets spanning a range of drug target classes. Both 2DA_Sign and 3DA_Sign significantly increase model performance across all datasets when compared with traditional 2DA and 3DA. Lastly, we find that limiting 3DA_Sign to maximum atom pair distances of 6 Å instead of 12 Å further increases model performance, suggesting that conformational flexibility may hinder performance with longer 3DA descriptors. Consistent with this finding, limiting the number of bonds in 2DA_Sign from 11 to 5 fails to improve performance.


In vitro characterizations of mesoporous hydroxyapatite as a controlled release delivery device for VEGF in orthopedic applications.
Following bone implant surgery, prolonged ischemic conditions at the implant site often result in postsurgical complications like failure of osseointegration at the bone-implant interface which can lead to implant failure. Thus, restoration of the vascular supply is paramount to the proper development of the bone. High surface area mesostructured materials have been shown to be attractive candidates for bone regeneration to enhance cell adhesion and cell proliferation. This study uses hydroxyapatite, a naturally occurring mineral in the bone, fabricated to a range of suitable pore sizes, infused with vascular endothelial growth factor (VEGF), to be progressively released to stimulate revascularization. In this study, several characterizations including nitrogen adsorption analysis, Fourier-transformed infrared spectroscopy, X-ray diffraction, field emission scanning electron microscope, and transmission electron microscope were used to evaluate the synthesized mesoporous hydroxyapatite (MHA). The results showed that MHA can gradually release VEGF for enhancing revascularization, which is beneficial for orthopedic applications.


The kinetics of substrate-induced inactivation.
The kinetics of a branched-pathway mechanism for a simple enzymic reaction were studied. In this mechanism there is reversible formation of an inactive form of the second complex along the pathway. This substrate-induced inactivation typically results in the progress curve showing a burst. Three parameters can be obtained from the progress curve: the initial rate, the final rate and the rate constant characterizing the transient. The rate constant for the conversion of the inactive form of the complex into the active form can be obtained either from these parameters or by measuring the regain of enzymic activity. The partition ratio can also be obtained from the three parameters; this is the ratio of the rate of conversion of complex into product to the rate of conversion of complex into inactive form. Simulations give guidance to the conditions required for accurate determinations of the rate constants.


Cloning of a novel antifungal promoter from Phaseolus vulgaris and the determination of its activity in stably transformed Nicotiana tabacum plants.
To investigate the transcriptional regulation of gene expression, chimeric fusions, between the beta-glucuronidase reporter gene (GUS) and the isolated promoter regions of the pvPDF gene (pvPDF-PRO: GUS), were constructed and introduced into Nicotiana tabacum. Analysis of transgenic pvPDF-PRO:GUS tobacco plants indicated that GUS activity was observed with all the promoter constructs with the strongest being in leaf followed by stem and roots. These results clearly demonstrate that pvPDF-PRO is a strong inducible and near-constitutive promoter and emphasize the great application potential for plant genetic engineering studies. Interestingly, a search for putative cis-acting elements in the pvPDF promoter architecture revealed the presence of some important transcription regulatory elements including: CAAT Box, TATA Box, CATA Box, and light regulatory elements (CCA1, GATA, GT-1). Taken together, these results further our understanding of the regulation of the pvPDF promoter activity.


Meiotic mutations in rye Secale cereale L.
Spontaneous meiotic mutations of winter rye Secale cereale L. (2n = 14) were revealed in inbred F2 progenies, which were obtained by self-pollination of F1 hybrids resulting from crosses of individual plants of cultivar Vyatka or weedy rye with plants of self-fertile inbred lines. The mutations cause partial or complete sterility, and are maintained in heterozygote condition. Six types of mutations were distinguished as the result of cytological analysis of meiosis and genetic analysis. (1) Plants with nonallelic asynaptic mutations sy1 and sy9 lacked bivalents in 96.8 and 67.0% metaphase I cells, respectively, formed only axial elements but not the mature synaptonemal complex (SC), and had defects in telomere clustering in early prophase I. (2) Weak asynaptic mutant sy3 showed incomplete synapsis at the start of SC degradation at diplotene and lower chiasma number; yet only 2% meiocytes lacked bivalents in MI. (3) Mutations sy2, sy6, sy7, sy8, sy10, and sy19 caused nonhomologous synapsis; i.e., a varying number of univalents and occasional multivalents were observed in MI, which was preceded by switches of pairing partners and fold-back synapsis at mid-prophase I. (4) Mutation mei6 led to the formation of protrusions and minor branched structures of the SC lateral elements. (5) Allelic mutations mei8 and mei8-10 caused irregular chromatin condensation along the chromosome length in prophase I, which was accompanied by chromosome sticking and fragmentation in MI. (6) Allelic mutations mei5 and mei10 determined chromosome supercondensation, caused the disturbance of meiotic spindle assembly, arrested meiosis at various stages but did not affect formation of the pollen wall, thus arrested meiocytes got covered with the pollen wall. Analysis of double mutants revealed recessive epistatic interactions for some mutations; the epistatic group was sy9 > sy1 > sy3 > sy19. This reflects the sequence of meiotic events controlled by the corresponding genes. The expression of sy2 and sy19 proved to be modified by additional genes. Most meiotic mutations found in rye have analogs in other plants.


[Biological availability of diagoxin in patients with or without gastric resection (Billroth II) (author's transl)].
Biological availability of digoxin tablets was measured during maintenance therapy in ten hospitalized patients who had had a Billroth II gastric resection at least two years previously. Twelve patients on digoxin maintenance for heart failure but without gastro-intestinal disease served as controls. The mean value of daily digoxin urinary excretion over ten days in the resection group was 38.09 +/- 0.70% of the administered dose. The serum-digitalis level 12 and 24 hours after the last dose of glycoside (0.5 mg) was 1.30 +/- 0.04 ng/ml and did not significantly differ from that of the control group, nor did digoxin elimination in urine and the digoxin/creatinin excretion ratio. It is concluded that two-third gastric resection with exclusion of antigrade duodenal passage does not influence biological availability of digoxin given in tablets.


Limited capsular release and controlled manipulation under anaesthesia for the treatment of frozen shoulder.
In light of recent interest in the cost-effectiveness of the treatment options available for frozen shoulder, we aimed to determine the results of limited anterior capsular release and controlled manipulation under anaesthesia (MUA) in the treatment of primary frozen shoulder in terms of patient-related outcomes measure, range of motion and re-intervention rates. This single-surgeon series included prospectively collected data on all patients undergoing capsular release with MUA from March 2011 until June 2013, with a minimum follow-up of 6 months from the index procedure. Outcome measures included pre- and postoperative Oxford Shoulder Score (OSS), range of motion and need for re-intervention. Fifty-four procedures were performed in 52 patients. Mean age 50 years (range 42 years to 59 years); male: female ratio = 11: 41. There was a highly statistically significant improvement in both pain and function modules of the OSS (p < 0.005) and range of motion (p < 0.005) at 6 months. The median postoperative score was 41 from a maximum of 48 points, with an average mean improvement of 24 points. Seventeen patients were diabetics. There was no significant difference in pre-operative and postoperative OSS or range of motion between the diabetic group and the non-diabetic groups. No patients required surgical re-intervention. A combination of limited capsular release and MUA for the treatment of primary frozen shoulder is a safe and effective procedure resulting in marked improvement in pain, function and range of motion.


Pulse control in a wide frequency range for a quasi-continuous wave diode-pumped cesium atom vapor laser by a pump modulation in the spectral domain.
Diode-pumped alkali-atom laser (DPAL) has attracted intense attention due to its inherently high quantum efficiency, a good beam quality, and a high potential in the power scaling. However, most of DPAL research has been confined to the continuous wave and only a few pulsed operations have been attempted with limited performances. Here, we proposed and experimentally demonstrated a new scheme using a fast mode-hopping in the pump laser diode (LD), which enabled the quasi-continuous-wave (QCW) pulse modulation in a cesium (Cs) DPAL to control both the pulse width and the repetition rate. The pump wavelength was efficiently modulated in a fast cycle within discrete spectral ranges provided by the mode-hopping in the pump LD. The spectral range was successfully adjusted to include the resonant D2 absorption line of Cs atom to result in an effective gain modulation. Using this proposed scheme, we successfully achieved Cs-DPAL QCW modulation, whose pulse width was varied from tens of microseconds to a few milliseconds and the repetition rate was also variable in a wide frequency range from 10 Hz to 7.0 kHz. Detailed pump modulation method and the corresponding laser characteristics are discussed. The proposed method can be readily applied to pulse modulation of other types of alkali vapor lasers overcoming the previous limitations of DPAL to further expand applications in various light-matter interactions.


[Prolonged neonatal cholestasis: prospective study].
Due to the urgency in choosing either clinical treatment or immediate surgical intervention, the study of the prolonged neonatal cholestasis involves two basic aims: the differential diagnosis between biliary atresia and neonatal hepatitis and the research into the associated etiological agents. So, in a prospective trial carried out in the 70's, 77 children with prolonged neonatal cholestasis were studied in order to establish the differential diagnosis between biliary atresia and neonatal hepatitis, followed by the evaluation of 108 children towards a pathogenesis of the prolonged neonatal cholestasis. The results of the differential diagnosis showed that within 18 items examined only 8 proved to be good biliary atresia indicators. They are as follows (in decreasing order): ductular proliferation (portal tracts), fibrosis (portal tracts), cholestasis (portal tracts), stools colour--acholia, hepatomegaly, canalicular cholestasis (lobule), infiltrate (portal tracts), giant cells (lobule). These eight items were then gathered in a sole indicator of great discriminative power, with a confidence level of 99%. The figures regarding the pathogenesis are: rubella virus 0%, herpes simplex virus 0%, listeriosis 0%, cytomegalovirus 2.2%, hepatitis B virus 2.4%, toxoplasmosis 2.8%, alpha-1-antitrypsin deficiency 13.1%, syphilis 21.1%, autoantibodies against the liver 58.4%. Such work thus revealed that those eight most important factors when differentiating biliary atresia from neonatal hepatitis remain as fundamental indicators and, when employed alongside other diagnostic methods, can help in the assembling of a multifactorial strategy less and less invasive and more precise. The pathogenic study, with its heavy dependency on time and place, has become more complete with the introduction of new diagnostic methods, evolving to the ideal progressive reduction of idiopathic processes.


Interactions between tacrolimus and antimicrobial agents.
Tacrolimus is being increasingly used as an immunosuppressive agent in transplant recipients. The drug is metabolized by the cytochrome P-450 3A system, thus antimicrobials that inhibit or induce these enzymes can alter levels of tacrolimus in the bloodstream. Tacrolimus is potentially nephrotoxic and neurotoxic; the likelihood of toxicity rises as blood levels of the drug increase. Increased tacrolimus levels and subsequent toxicity have been produced by a number of antimicrobial agents that inhibit the cytochrome P-450 3A system. Conversely, drugs with the potential to induce the cytochrome P-450 3A system can reduce the levels of tacrolimus in the blood, leading to increased risk of acute rejection in transplant recipients. Antimicrobial agents that can have adverse effects on renal function may add to the nephrotoxicity of tacrolimus. The potential for drug interactions should be reviewed before antimicrobial agents are prescribed to patients treated with tacrolimus.


A probiotic intervention on pain hypersensitivity and microbiota composition in patients with osteoarthritis pain: Study protocol for a randomized controlled trial.
This study aims to examine the effects of probiotics on pain hypersensitivity at the end of a six-week intervention program in patients with osteoarthritis (OA)-related pain. This double-blind randomized controlled clinical trial with two parallel arms will be conducted between January 2021 and July 2022. At least 30 participants (age range, 50 to 90 years) of both sexes with a diagnosis of symptomatic hip or knee (Kellgren-Lawrence scale ≥3) will be recruited in each arm (total n=60) to achieve adequate statistical power in the analyses. The intervention will be administered for six weeks followed by a four-week follow-up period. The experimental group will receive a probiotic product plus the usual medical care. The control group will receive a probiotical sham plus the usual medical care. Assessment points will be measured at baseline, end of intervention, and one-month post-intervention. The outcomes of this intervention will be a change in visual analog scale pain and the gut microbiota composition. Group by time effects will be compared using mixed-model analysis of variance. A reduction in pain hypersensitivity in patients with knee OA-related pain could suggest an involvement of microbiota, or part of it, in chronic pain state mechanisms.


A common variant in pre-miR-146 is associated with coronary artery disease risk and its mature miRNA expression.
miRNAs are small non-coding RNAs that play an important role in numerous physiological processes. Common single nucleotide polymorphisms (SNPs) in pre-miRNAs may change their property through altering miRNAs expression and/or maturation, resulting in diverse functional consequences. To date, the role of genetic variants in pre-miRNAs on coronary artery disease (CAD) risk remains poorly understood. Here we aimed to evaluate the influence of three common SNPs in pre-miRNAs (miR-146a rs2910164 G>C, miR-196a2 rs11614913 C>T, miR-499 rs3746444 T>C) on individual susceptibility to CAD in a Chinese population of 295 CAD patients and 283 controls. Genotyping was performed using polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) method. In a logistic regression analysis, we detected an association of rs2910164 in pre-miR-146a with the CAD risk; compared with the GG homozygotes, the GC heterozygotes [odds ratio (OR)=1.89, 95% confidence interval (CI)=1.06-3.36, P=0.029] and the CC homozygotes (OR=1.83, 95% CI=1.01-3.32, P=0.046) genotype were statistically significantly associated with the increased risk for CADs. As we used further genotype association models, we found a similar trend of the association in recessive model (OR=1.86, 95% CI=1.09-3.19, P=0.023). We also found that the genotypes of miR-146a rs2910164 were associated with its mature miRNA expression by analyzing 23 PBMC samples from CAD patients. Individuals carrying rs11614913 GC or CC genotypes showed 3.2-fold higher expression compared to GG genotype carriers (P<0.05). We observed no association of the other two SNPs in miR-196a2 (rs11614913) and miR-499 (rs3746444) with the CAD incidence. Our data provide the first evidence that the miR-146a rs2910164 polymorphism is associated with increased risk of CAD in Chinese Han population, which may be through influencing the expression levels of the miRNA.


How to transect the liver? A history lasting more than a century.
There is a close relationship between blood loss during transection and unfavorable outcome. Many different methods have been used in order to cut the parenchyma, while leaving vital structures intact, coagulate small vessels and seal small biliary ducts. The first method described was the finger-fracture technique and, alternatively, the clamp-crushing method using a small forceps. With this technique, the liver is crushed between the 'jaws', and the vessels and bile ducts are successively ligated and divided. Technological research using different sources of energy developed the water jet dissectors and the ultrasonic dissectors. The CUSA® has been widely adopted for the fascinating way it could selectively destroy and aspirate parenchyma leaving vascular structures almost intact. Several studies have been addressed to clarify these critical points. However, in the majority of cases they are underpowered to demonstrate clear advantages of one method over the others. In conclusion, the evidence suggested no superiority of other techniques over clamp-crushing. But it must be taken into account that it requires strictly hepatic pedicle clamping. The devices available should be used within the limits of each instrument, as well as the surgical skills of the surgeon. Probably the best option should be a combined approach.


Analysis of Serum Cytokine Profile in Pemphigus.
Pemphigus is a group of autoimmune blistering diseases affecting skin and mucous membranes. While pemphigus is an autoantibody mediated disease, the role of T cells and cytokines in the pathogenesis is being increasingly recognized. This study was conducted to observe alterations in the serum cytokine levels of patients with pemphigus vulgaris (PV), pemphigus foliaceous (PF), paraneoplastic pemphigus (PNP) and compare with bullous pemphigoid (BP) and healthy subjects. A total of 75 subjects (28 PV, 13 PF, 7 PNP, 7 BP, and 20 healthy controls) were included, all patients in active disease state. Serum levels of interferon (IFN)-γ, interleukin (IL)-4, IL-6, IL-17A, IL-10, tumor necrosis factor (TNF)-α, and IL-8 were measured by enzyme-linked immunosorbent assay. The median concentration of IFN-γ was lower in PV and BP patients compared to control (0.77, 0.34 and 1.63 pg/ml, respectively). IL-6 and IL-10 was significantly higher in PNP patients compared to control (4.92 and 0.24 pg/ml for IL-6, 0.86 and <0.12 pg/ml for IL-10, respectively). IL-8 was increased significantly in PV and PNP patients compared with control (11.85, 31.5 and 8.31 pg/ml, respectively). For IL-4, IL-17A and TNF-α, no significant difference was observed between the five groups. The decreased level of IFN-γ in PV may imply suppressed Th1 response in the active disease stage. A Th2 predominant response is suggested in the active stage of PNP, with elevated serum levels of IL-6 and IL-10. Increased level of proinflammatory cytokine IL-8 is observed in the sera of PV and PNP patients.


Health Services Use Among Jailed Women with Alcohol Use Disorders.
Correctional facilities now house unprecedented numbers of women with complex treatment needs. This investigation applied the Behavioral Model for Vulnerable Populations to study 168 jailed women with alcohol use disorders. It described the sample's predisposing (age, race, victimization), enabling (health insurance), and need (self-reported medical, substance use, and mental health problems) factors and examined associations of these factors with pre-incarceration services utilization. Most participants had clinically significant levels of depression and PTSD symptoms, most took psychiatric medications, and most had been victimized. Participants reported considerable health services utilization. Younger, Black, and uninsured women utilized fewer medical and mental health services. Drug use was associated with less use of medical services, but more use of alcohol and drug services. High rates of health services use support the need for integrated, ongoing care for substance-using women before, during, and after incarceration.


[Epidemiological analysis of suicide in Brazil from 1980 to 2006].
The objective of this study was to carry out an epidemiological analysis of the suicide rates found in Brazilian regions and state capitals between 1980 and 2006. Data on mortality rates due to suicide were collected from the Departamento de Informática do Sistema Unico de Saúde (Information Technology Department of the Brazilian Public Health System--DATASUS). A total of 158,952 suicide cases were registered between 1980 and 2006, excluding those cases in which the individual was less than 10 years old (n = 68). In the period under study, the total suicide rate in Brazil increased from 4.4 to 5.7 deaths per 100,000 inhabitants (29.5%). Higher average rates were found in the South (9.3) and Central-West (6.1) regions. Men were more likely to commit suicide. The highest suicide rates were found in the 70-years or above age range while the highest increases were found in the 20 to 59 year age bracket. The most dominant social-demographic characteristics of the persons who committed suicide in the period under study were low educational level and singlehood. The most common methods of suicide were hanging, fire arms and poisoning. Although in Brazil the rate increased 29.5% in 26 years, the national rate is still considered to be low when compared to worldwide suicide rates (average of 4.9 per 100,000 inhabitants). Suicide rates in Brazilian regions vary broadly, ranging from 2.7 to 9.3.


Discriminant function sexing of the calcaneus of the South African whites.
The skull and some postcranial elements, such as the humerus, femur, and tibia, have been used in their intact states for sex determination in forensic and archaeological cases. But, in practice, these bones are often recovered in fragmented states, which render them unsuitable for use in sex determination. The calcaneus is a compact bone that is able to withstand high tensile forces. Some of its parameters have been used for sex determination in American whites and blacks (1) and Italians (2). This bone has not been used for sex determination in the South African white population. Therefore, the aim of this study was to assess the degree of sexual dimorphism of the calcaneus of the South African white population sample, derive discriminant function score equations for use in sex determination, and determine the level of accuracy of its sex-determining ability. Nine parameters were measured on each pair of 53 male and 60 female calcanei of known South African white skeletons, obtained by a random sampling technique from the Raymond A. Dart Collection of Human Skeletons, School of Anatomical Sciences, University of the Witwatersrand, Johannesburg. Basic statistic and discriminant function analysis was performed on the acquired data. The basic statistics showed that all measured parameters were sexually dimorphic. Discriminant function score equations were generated for use in sex determination. The average accuracy of sex classification ranged from 73 to 86% for the univariate method, 81 to 91% for the stepwise method, and 82 to 92% for the direct method. It is concluded that the calcaneus is useful for sex determination in the South African white population.


A novel copper-responsive regulon in Mycobacterium tuberculosis.
In this work we describe the identification of a copper-inducible regulon in Mycobacterium tuberculosis (Mtb). Among the regulated genes was Rv0190/MT0200, a paralogue of the copper metalloregulatory repressor CsoR. The five-locus regulon, which includes a gene that encodes the copper-protective metallothionein MymT, was highly induced in wild-type Mtb treated with copper, and highly expressed in an Rv0190/MT0200 mutant. Importantly, the Rv0190/MT0200 mutant was hyper-resistant to copper. The promoters of all five loci share a palindromic motif that was recognized by the gene product of Rv0190/MT0200. For this reason we named Rv0190/MT0200 RicR for regulated in copper repressor. Intriguingly, several of the RicR-regulated genes, including MymT, are unique to pathogenic Mycobacteria. The identification of a copper-responsive regulon specific to virulent mycobacterial species suggests copper homeostasis must be maintained during an infection. Alternatively, copper may provide a cue for the expression of genes unrelated to metal homeostasis, but nonetheless necessary for survival in a host.


Transcriptomic analysis of cell-free fetal RNA suggests a specific molecular phenotype in trisomy 18.
Trisomy 18 is a common human aneuploidy that is associated with significant perinatal mortality. Unlike the well-characterized "critical region" in trisomy 21 (21q22), there is no corresponding region on chromosome 18 associated with its pathogenesis. The high morbidity and mortality of affected individuals has limited extensive investigations. In order to better understand the molecular mechanisms underlying the congenital anomalies observed in this condition, we investigated the in utero gene expression profile of second trimester fetuses affected with trisomy 18. Total RNA was extracted from cell-free amniotic fluid supernatant from aneuploid fetuses and euploid controls matched for gestational age and hybridized to Affymetrix U133 Plus 2.0 arrays. Individual differentially expressed transcripts were obtained by two-tailed t tests. Over-represented functional pathways among these genes were identified with DAVID and Ingenuity(®) Pathways Analysis. Results show that three hundred and fifty-two probe sets representing 251 annotated genes were statistically significantly differentially expressed between trisomy 18 and controls. Only 7 genes (2.8% of the annotated total) were located on chromosome 18, including ROCK1, an up-regulated gene involved in valvuloseptal and endocardial cushion formation. Pathway analysis indicated disrupted function in ion transport, MHCII/T cell mediated immunity, DNA repair, G-protein mediated signaling, kinases, and glycosylation. Significant down-regulation of genes involved in adrenal development was identified, which may explain both the abnormal maternal serum estriols and the pre and postnatal growth restriction in trisomy 18. Comparison of this gene set to one previously generated for trisomy 21 fetuses revealed only six overlapping differentially regulated genes. This study contributes novel information regarding functional developmental gene expression differences in fetuses with trisomy 18.


Antihyperglycemic Activity of Petroleum Ether Leaf Extract of Ficus krishnae L. on Alloxan-induced Diabetic Rats.
The petroleum ether leaf extract of Ficus krishnae has been evaluated for the management of diabetes in alloxan induced diabetic rats. Phytochemical screening of the leaf extract for various chemical compounds has also been carried out. Leaf extract was administered continuously for 21 days orally at a dose of 200 mg/kg. Along with this, the blood glucose level was monitored at regular intervals to understand the activity of the extract. The leaf extract has decreased the blood glucose level of diabetic rats which was comparable to an antidiabetic standard drug, glibenclamide, given at a dose of 2.5 mg/kg. It has been observed that the leaves of Ficus krishnae possess antidiabetic activity and it reduces the blood glucose level significantly. The phytochemical screening of leaf has revealed the presence of carbohydrates, flavonoids, tannins, glycosides, terpenoids, steroids, alkaloids, gums and mucilage, phlobatannins, reducing sugars and phenolic compounds. The Fourier Transform Infrared analysis of glibenclamide and leaf powder has displayed some common absorption spectra. This shows that leaf powder has a molecule which is close to glibenclamide. Wavelength Dispersive X-Ray Fluorescence spectroscopy have shown the presence of cellulose, Ca, Si, K, Cl, Mg, P, S, Al, Fe, Na, Sr, Pd, Zn, Mn, Cr, Mo, Br, Ni, Rb and Zr. It is assumed that these elements alongwith other chemical compounds of the plant species may play a role in the management of diabetes. The Raman Specta of both glibenclamide and leaf powder has also shown some similarities. The results obtained during the present investigation have revealed the antidiabetic activity of Ficus krishnae leaves. The phytochemical screening has indicated the various chemical constituents likely to be responsible for this activity. The Fourier Transform Infrared, Wavelength Dispersive X-Ray Fluorescence and Raman Specta of the leaf powder suggested that there is some glibenclamide like molecule or its derivatives which is imparting antidiabetic activity.


Genetic and molecular characterization of Tn21, a multiple resistance transposon from R100.1.
Tge transposon Tn21 has been transposed from R100.1 to plasmid pACYC184 and, from the resulting recombinants, to plasmid R388. The sites of insertion and the orientation of the element in several pACYC184::Tn21 recombinants have been examined. Restriction enzyme analysis of these recombinants has resulted in a detailed map of Tn21; this is compared with the published maps of the relevant part of R100.1. Heteroduplex analysis has shown short inverted repeat sequences at the ends of the element. With various in vitro-generated deletion mutants of Tn21, the internal gene necessary for transposition (tnpA) was localized within the terminal 4.3 kilobases of the right-hand end of the element. Genetic analysis of transposition of Tn21 suggests that the process proceeds via cointegrates. Since the end products of transposition are simple recombinants of the element and the recipient replicon, Tn21 must contain a gene that codes for a resolvase type of activity (tnpR gene).


Effect of role play education on primiparous women's fear of natural delivery and their decision on the mode of delivery.
The number of women who select cesarean section due to fear of childbirth has increased. Role play education seems to be a helpful method to remove or reduce the fear of childbirth. Therefore, this study aimed to investigate the effect of role play education on primiparous women's fear of natural delivery and their decision on the mode of delivery. In this blind clinical trial, 67 primiparous women with natural pregnancy at 34-36 weeks of gestational age and with no indication of cesarean section were selected from the health care centers in Mashhad. They were randomly assigned to two groups who underwent pre-test and post-test with the help of delivery attitude questionnaire to investigate their fear of childbirth and a researcher-made pregnant women's decision investigation questionnaire. Education through role play was conducted in the form of three scenarios during seven stages. The findings were analyzed by Fisher's exact test and independent t-test through SPSS. The two groups were significantly different concerning the fear of childbirth after the intervention (P = 0.007), and the fear score showed a higher reduction in the role play group compared to the lecture group. There was a significant difference between the two groups concerning the reduction of elective cesarean section and the decision on the mode of delivery at the time of admission in the labor room (P = 0.000). About 75% in the lecture group and 100% in the role play group selected natural delivery. In the present study, the effect of role play was more in making a decision on natural delivery, reducing the fear of childbirth, and reducing the rate of elective cesarean section. It is suggested to use role play method to educate pregnant women to reduce the rate of cesarean sections.


Neuroscience and heart-brain medicine: the year in review.
Important recent publications in the area of neuroscience and heart-brain medicine center largely around three topics: (1) mechanisms of cardiac sympathetic denervation in Parkinson disease, (2) cytoplasmic monoamine metabolites as autotoxins, and (3) the validity of power spectral analysis of heart rate variability to indicate cardiac sympathetic tone. Findings by Orimo et al support a centripetal, retrograde pathogenetic process involving alpha-synuclein deposition and degeneration of cardiac noradrenergic neurons in Parkinson disease. Several studies suggest that processes increasing cytoplasmic monoamines lead to neuronal loss from auto-oxidation or enzymatic oxidation. Lack of correlation between commonly used indices from power spectral analysis of heart rate variability and cardiac norepinephrine spillover casts doubt on the validity of power spectral analysis to indicate cardiac sympathetic tone.


Identification of molecular mechanisms related to nonthyroidal illness syndrome in skeletal muscle and adipose tissue from patients with septic shock.
Septic shock is one of various causes of nonthyroidal illness syndrome (NTIS). In humans, the molecular mechanisms involved in NTIS are mostly unknown. The aim of this study was to investigate, in patients with NTIS secondary to septic shock, changes in the expression of genes involved in the actions of thyroid hormones and in the activity of deiodinase enzymes, in two tissues important for protein and energy metabolism, skeletal muscle (SM) and subcutaneous adipose tissue (SAT). Hospitalized patients were divided into a control and a septic shock NTIS group. Serum collection for biochemical measurements, and SM and SAT biopsies for mRNA expression analysis of thyroid hormone receptors (THRB1, THRA1), retinoid X receptors (RXRA, RXRB, RXRG), nuclear receptor corepressor (NCOR1), silencing mediator of retinoid and thyroid hormone receptor (SMRT), steroid receptor coactivator (SRC1), type 1 and 2 deiodinases (D1, D2), monocarboxylate transporter 8 (MCT8), SECIS binding protein 2 (SBP2) and uncoupling protein 3 (UCP3) as well as D1, D2 and D3 enzyme activity measurements. The NTIS group had lower serum TSH, and free T3 and higher rT3 than controls. D1 and D3 were detected in SAT, with no differences found between the two groups; SM had very low D2 activity and again no differences were found between groups; D3 activity in SM was higher in NTIS than controls. SM expression of THRB1, RXRG and D2 was lower and RXRA higher in NTIS than controls. SAT from NTIS patients had lower MCT8, THRB1, THRA1, RXRG and SMRT, and higher UCP3 expression than controls. In patients with septic shock NTIS tissue responses are orientated to decrease production and increase degradation (muscle) or decrease uptake (adipose tissue) of T3, as well as to decrease thyroid hormone actions.


Chromosome-biased binding and gene regulation by the Caenorhabditis elegans DRM complex.
DRM is a conserved transcription factor complex that includes E2F/DP and pRB family proteins and plays important roles in development and cancer. Here we describe new aspects of DRM binding and function revealed through genome-wide analyses of the Caenorhabditis elegans DRM subunit LIN-54. We show that LIN-54 DNA-binding activity recruits DRM to promoters enriched for adjacent putative E2F/DP and LIN-54 binding sites, suggesting that these two DNA-binding moieties together direct DRM to its target genes. Chromatin immunoprecipitation and gene expression profiling reveals conserved roles for DRM in regulating genes involved in cell division, development, and reproduction. We find that LIN-54 promotes expression of reproduction genes in the germline, but prevents ectopic activation of germline-specific genes in embryonic soma. Strikingly, C. elegans DRM does not act uniformly throughout the genome: the DRM recruitment motif, DRM binding, and DRM-regulated embryonic genes are all under-represented on the X chromosome. However, germline genes down-regulated in lin-54 mutants are over-represented on the X chromosome. We discuss models for how loss of autosome-bound DRM may enhance germline X chromosome silencing. We propose that autosome-enriched binding of DRM arose in C. elegans as a consequence of germline X chromosome silencing and the evolutionary redistribution of germline-expressed and essential target genes to autosomes. Sex chromosome gene regulation may thus have profound evolutionary effects on genome organization and transcriptional regulatory networks.


Proline-rich vaccine candidate antigen of Coccidioides immitis: conservation among isolates and differential expression with spherule maturation.
A proline rich antigen (PRA), which protects mice against Coccidioides immitis, has been analyzed for differential expression and variation among isolates. Northern blots of RNA from different stages of growth were probed with previously cloned cDNA and showed that mRNA for PRA increased as spherules transformed and matured from mycelia. This pattern corresponds to the relative potency of whole cell vaccines from similar preparations. The PRA gene was then cloned from a genomic library of the Silveira strain of C. immitis and its sequence analyzed. Eight other coccidioidal isolates, selected for diversity in geographic origin and resulting clinical disease, demonstrated heterogeneity in Southern blots and in sequences of polymerase chain reaction products. Silveira differed from other California isolates at 33 of 555 bases, whereas it differed from non-California isolates by <=2 bases. Only one of these substitutions affected protein sequence. Thus, tests or vaccines based on this gene are likely to cover most isolates.


[Sonography in blunt abdominal trauma].
Ultrasonic examination was performed in 79 patients with blunt abdominal trauma, using 3.5 mHz real-time sonography. The examination aimed at clarifying intraperitoneal and retroperitoneal haemorrhage and injury to organs. Sonographic detection of haemorrhage succeeded in 13/13 of spleen ruptures and in one case of a liver rupture. 6 renal traumas were recognised by means of indirect signs and only twice by the direct identification of a band-shaped interruption of the parenchyma. Non-invasive sonography can be employed primarily in place of diagnostic peritoneal lavage for the detection of life-threatening intraperitoneal haemorrhages with an accuracy of more than 90%. Primary diagnosis of the source of haemorrhage is limited to individual cases because of the unfavourable conditions under which examination is performed. In unclear cases essential additional information is supplied by sonographic examination of the course of the disease.


Oral anticoagulation in the hospital: analysis of patients at risk.
Warfarin is one of the most commonly used medications associated with adverse events. Warfarin therapy is often initiated or continued in the hospital, yet hospitalization increases the risk of poor anticoagulation control with warfarin. To help understand this, we retrospectively reviewed the records of patients admitted to our hospital during a 6-month period who were given at least one dose of warfarin. To explore factors that may have contributed to poor anticoagulation control, we compared characteristics of patients with an international normalized ratio (INR) ≥ 5 at some point during hospitalization with those of a group of matched controls who also received warfarin and had INR <5. Among the 35 patients identified who had an INR ≥ 5, concomitant use of antibiotics was more common than among 105 matched controls; improper warfarin dosing also appeared to contribute to the high INRs. These findings indicate possible targets for intervention to improve patient safety.


[Relationships between emotional distress, empowerment perception and self-care behavior and quality of life in patients with type 2 diabetes].
Diabetes mellitus is a chronic disorder prevalent worldwide that, in addition to being costly to manage, severely impacts patient quality life. Therefore, it is extremely important to understand the factors associated with quality of life in diabetic patients. The purposes of this study were to investigate relationships between variables including demographics, disease characteristics, emotional distress, empowerment perception, and self-care behavior and quality of life, as well as to identify the important explanatory factors of quality of life in patients with type 2 diabetes. This study used a design that was cross-sectional, descriptive and correlated. A total of 220 patients diagnosed with type 2 diabetes in the previous ten year period were recruited as participants in this study. Data was collected using a structured questionnaire that incorporated scales to capture demographic, disease characteristic, emotional distress, empowerment perception, self-care behavior and quality of life data. The quality of life scale measured the three domains of satisfaction, impact, and worried about the diabetes. Results included: (1) the standardized score of quality of life was 79.6, considered in the moderate to high range; (2) Education level, age, "diabetes shared care" participation and insulin therapy were variables with significant impact upon quality of life; (3) The more severe the emotional distress, the worse quality of life scores were in every domain; (4) The better the perception of empowerment, the better the score in the satisfaction domain; (5) The better self care behavior was, the better the score in the satisfaction domain; and (6) emotional distress was the most important explanatory factor of quality of life, accounting for 28.7%-53.8% of total variance. The findings of this study suggested that health professionals should evaluate emotional distress of diabetic patients in the early stage, provide emotional support and consultation and apply empowerment strategy to promote their quality of life.


Remote Estimation of Peripheral Oxygen Saturation and Pulse Rate From Facial Analysis Using a Smartphone Camera.
Vital sign monitoring is an invaluable tool for healthcare professionals, both in the hospital and at home. Traditional measurement devices provide accurate readings but require physical contact with the patient which often is unsuitable, furthermore contact-based devices have been reported to fail by loosing contact due to movement as severe events occur, therefore, a contactless method is necessary.We hypothesize that, in ideal scenarios, it is possible to estimate both SpO2 and pulse rate using only facial video recorded with a smartphone's front-facing camera. To test this hypothesis, a dataset of 10 healthy subjects performing various breathing patterns while being recorded with a smartphone camera was collected during ideal lighting conditions.Using advanced image and signal processing methods to acquire remote photoplethysmography (rPPG) estimates from a patient's forehead, our proposed method can achieve SpO2 estimation results with Arms = 1.34% (accuracy RMS) and MAE ± STD = 1.26 ± 0.68% (mean average error) across a SpO2 range of 92% to 99% (percentage point SpO2) and pulse rate estimation results with Arms = 3.91 bpm (beats per minute) and MAE ± STD = 3.24±2.11 bpm across a pulse rate range of 60 bpm to 90 bpm. We conclude from these results, that remote vital sign estimation using facial videos recorded entirely with a smartphone camera is possible.


Syntheses of alternating oligo-2'-O-methylribonucleoside methylphosphonates and their interactions with HIV TAR RNA.
Oligonucleotide analogues 15-20 nucleotides in length have been prepared, whose sequences are complementary to nucleotides in the upper hairpin of HIV TAR RNA. These alternating oligonucleoside methylphosphonates, mr-AOMPs, contain 2'-O-methylribonucleosides and alternating methylphosphonate and phosphodiester internucleotide linkages. The methylphosphonate and phosphodiester linkages of these oligomers are highly resistant to hydrolysis by exonuclease activity found in mammalian serum and to endonucleases, such as S1 nuclease. The oligomers were prepared using automated phosphoramidite chemistry and terminate with a 5'-phosphate group, which provides an affinity handle for purification by strong anion exchange HPLC. A 15-mer mr-AOMP, 1676, that is complementary to the 5'-side of the TAR RNA hairpin, including the 3-base bulge and 6-base loop region, forms a 1:1 duplex with a complementary RNA 18-mer, mini-TAR RNA. The T(m) of this duplex is 71 degrees C, which is similar to that of the duplex formed by the corresponding all phosphodiester 15-mer. Introduction of two mismatched bases reduces the T(m) by 17 degrees C. The apparent dissociation constant, K(d), for the 1676/mini-TAR RNA duplex as determined by an electrophoretic mobility shift assay at 37 degrees C is 0.3 nM. Oligomer 1676 also binds tightly to the full length TAR RNA target under physiological conditions (K(d) = 20 nM), whereas no binding was observed by the mismatched oligomer. A 19-mer that is complementary to the entire upper hairpin also binds to TAR RNA with a K(d) that is similar to that of 1676, a result that suggests only part of the oligomer binds. When two of the methylphosphonate linkages in the region complementary to the 6-base loop are replaced with phosphodiester linkages, the K(d) is reduced by approximately a factor of 10. This result suggests that interactions between TAR RNA and the oligomer occur initially with nucleotides in the 6-base loop, and that these interactions are sensitive to presence and possibly the chirality of the methylphosphonate linkages in the oligomer. The high affinities of mr-AOMPs for TAR RNA and their resistance to nuclease hydrolysis suggests their potential utility as antisense agents in cell culture.


Reduced susceptibility in laboratory-selected mutants of Aspergillus fumigatus to itraconazole due to decreased intracellular accumulation of the antifungal agent.
To study the mechanism of resistance of Aspergillus fumigatus to itraconazole, spontaneous mutants with reduced susceptibility were selected by spreading 2 x 10(8) conidia from a clinical isolate (W73355) susceptible to miconazole (MIC 2 mg/l), itraconazole (MIC 0.25 mg/l) and amphotericin B (MIC 0.5 mg/l) on 40 peptone yeast extract glucose agar plates containing miconazole (32 mg/l). The 19 colonies that grew (frequency 0.95 x 10(-7)) in the presence of miconazole were screened by broth macrodilution technique for their susceptibility to itraconazole. A total of two isolates (frequency 1 x 10(-8)) MCZ14 and MCZ15 had MICs of 16 and 8 mg/l, respectively, for itraconazole. Both MCZ14 and MCZ15 showed concomitant increases in MICs for ketoconazole and miconazole, but not for amphotericin B. Growth inhibition studies as well as kill curve experiments revealed that MCZ14 and MCZ15 were less susceptible to itraconazole compared to the parental strain. The intracellular accumulation of itraconazole in A. fumigatus was time and concentration dependent. Maximum accumulation was obtained within 30 min at 5 microM itraconazole. In MCZ14 and MCZ15 intracellular accumulation of [3H]itraconazole was reduced by approximately 80 and 60%, respectively, compared to the susceptible parent. The respiratory inhibitor carbonyl cyanide m-chlorophenyl hydrazone at 200 microM reduced the intracellular accumulation of itraconazole by approximately 36.2% (P < or = 0.05) in the parent and in the mutant strains. These results suggest that (i) the reduced accumulation of itraconazole in MCZ14 and MCZ15 is due to diminished permeability of the drug and perhaps not due to efflux, (ii) the uptake of itraconazole in A. fumigatus may be an energy dependent process, and (iii) decreased accumulation of itraconazole is at least in part responsible for the reduced susceptibility of the mutant isolates to itraconazole.


Personalized AAC Intervention to Increase Participation and Communication for a Young Adult with Down Syndrome.
Many adults with Down syndrome (DS) experience difficulty with speech production, and intelligibility challenges and communication breakdowns are common. Augmentative and alternative communication (AAC) intervention can provide important supports for persons with these complex communication needs, but must be customized to address the goals, strengths, and needs of the individual. This article provides a description of a personalized AAC intervention for a young adult with DS whose speech was frequently unintelligible. The AAC intervention made use of a video visual scene display (VSD) approach, and was investigated in two separate studies in two key community settings: An inclusive post-secondary education program (a single-case reversal ABAB design), and a community shopping activity (a non-experimental AB case study design). The participant demonstrated sharp increases in successful communication and participation in both settings following the introduction of the video VSD, and both the participant and key stakeholders viewed the intervention positively. The results provide preliminary evidence that personalized AAC intervention, including the use of a video VSD approach, can provide important supports for communication and participation in community settings for adults with DS and complex communication needs.


Interpretation of high-resolution current source density profiles: a simulation of sublaminar contributions to the visual evoked potential.
Current source density (CSD) analysis provides an index of the location, direction, and density of transmembrane currents that arise with synchronous activation of neural tissue and that generate an evoked potential profile in the extracellular medium. In neocortex and other laminated structures, a simplified, one-dimensional CSD analysis can be computed by differentiation of voltages sampled at discrete points in a linear array. One-dimensional CSD analysis is a practical and accurate method for defining both regional activity patterns and neural generators of surface-recorded evoked and event-related potentials. In computing the CSD, common practices of differentiating across spatial grids of 200 microns or more and use of spatial smoothing routines help to reduce noise, but severely limit the spatial resolution available to the analysis. High-resolution CSD procedures (i.e., 3 point differentiation using a spatial grid of 100 microns or less) are more suited to identification of processes within individual cortical laminae or sublaminae, but can magnify the contributions of computational artifacts. Despite the inclusion of independent indices of cellular activity (e.g., multiunit activity), both high- and low-resolution analyses may indicate current source and sink configurations for which there is more than one plausible physiological interpretation. In the present study we examined the resolving capacity and pitfalls of common CSD procedures using simulated ensembles of current dipoles. These were positioned and oriented to model the depolarization of lamina 4C stellate cells and thalamocortical afferents in macaque striate cortex. Empirically, the surface N40 appears in association with a CSD configuration which includes current sinks within the thalamorecipient (stellate) subdivisions of lamina 4C and a large current source extending considerably below 4C. Dipole ensemble contributions to the CSD profile were computed and compared to physiological data from this region. Small asymmetries in activation of model stellate laminae were sufficient to produce substantial open field contributions. However, the best fit with empirical CSD profile was found when the simulation included contributions from thalamocortical axons, along with both open and closed field contributions from dual stellate cell sublaminae. High-resolution CSD profiles were shown to be interpretable when computational artifacts characteristic of closed and open fields were identified using a series of differentiation grids.


Clinical predictors of early left ventricular thrombus formation in acute myocardial infarction.
Clinical epidemiological and echocardiographic risk factors relating to the development of a left ventricular thrombus were studied retrospectively in patients with acute myocardial infarction. The data on 1833 consecutive patients treated for acute myocardial infarction during a 10-year period were processed retrospectively. Transthoracic echocardiography was performed on each patient 65.0 +/- 5.5 hours after hospital admission. A left ventricular thrombus was detected in 145 patients (7.9%). The patients with acute myocardial infarction and a left ventricular thrombus had significantly lower frequencies of 1) myocardial infarction in their family history (3% versus 11%, respectively), 2) hospital admission within 24 hours from the onset of chest pain (17% versus 50%, respectively), 3) thrombolytic therapy (8% versus 23%, respectively) and 4) current smoking (24% and 35%, respectively) than those without a left ventricular thrombus. In contrast, anterior infarction (81% versus 38%, respectively), left ventricular dilatation (30% versus 19%, respectively), dyskinesis of the left ventricular wall (23% versus 10%, respectively), an aneurysm (22% versus 7%, respectively) and a reduced systolic left ventricular function (ejection fraction < 40%) (28% versus 17%, respectively) were more frequent in the presence of a left ventricular thrombus after myocardial infarction. Multivariate analysis of the results revealed that the presence of anterior myocardial infarction and an aneurysm is associated with significantly increased hazard ratios. On the other hand, early hospitalization and a positive family history of infarction significantly lowered the hazard ratio. The frequency of a left ventricular thrombus was significantly higher in spring and winter. The results presented in this paper confirm the significant hazard of certain parameters [location of infarction (anterior) and aneurysm] as concerns left ventricular thrombus formation among patients with acute myocardial infarction. Early hospitalization was found to lower the risk of thrombus formation. These echocardiographic and clinical parameters may be useful in the establishment of the individual risk of intracavital thrombus formation and may be of help in everyday medical practice.


Identifying Frameworks for Validation and Monitoring of Consensual Behavioral Intervention Technologies: Narrative Review.
Changing health behaviors, such as smoking, unhealthy eating, inactivity, and alcohol abuse, may have a greater impact on population health than any curative strategy. One of the suggested strategies is the use of behavioral intervention technologies (BITs). They open up new opportunities in the area of prevention and therapy and have begun to show benefits in the durable change of health behaviors in patients or those at risk. A consensual and international paradigm was adopted by health authorities for drugs 50 years ago. It guides their development from research units to their authorization and surveillance. BITs' generalization brings into question their upstream evaluation before being placed on the market and their downstream monitoring once on the market; this is especially the case in view of the marketing information provided by manufacturers and the scarcity and methodological limits of scientific studies on these tools. This study aims to identify and categorize the frameworks for the validation and monitoring of BITs proposed in the literature. We conducted a narrative literature review using MEDLINE, PsycINFO, and Web of Science. The review items included the following: name, publication year, name of the creator (ie, first author), country, funding organization, health focus, target group, and design (ie, linear, iterative, evolutive, and/or concurrent). The frameworks were then categorized based on (1) translational research thanks to a continuum of steps and (2) the three paradigms that may have inspired the frameworks: biomedical, engineering, and/or behavioral. We identified 46 frameworks besides the classic US Food and Drug Administration (FDA) five-phase drug development model. A total of 57% (26/46) of frameworks were created in the 2010s and 61% (28/46) involved the final user in an early and systematic way. A total of 4% (2/46) of frameworks had a linear-only sequence of their phases, 37% (17/46) had a linear and iterative structure, 33% (15/46) added an evolutive structure, and 24% (11/46) were associated with a parallel process. Only 12 out of 46 (26%) frameworks covered the continuum of steps and 12 (26%) relied on the three paradigms. To date, 46 frameworks of BIT validation and surveillance coexist, besides the classic FDA five-phase drug development model, without the predominance of one of them or convergence in a consensual model. Their number has increased exponentially in the last three decades. Three dangerous scenarios are possible: (1) anarchic continuous development of BITs that depend on companies amalgamating health benefits and usability (ie, user experience, data security, and ergonomics) and limiting implementation to several countries; (2) the movement toward the type of framework for drug evaluation centered on establishing its effectiveness before marketing authorization to guarantee its safety for users, which is heavy and costly; and (3) the implementation of a framework reliant on big data analysis based on a posteriori research and an autoregulation of a market, but that does not address the safety risk for the health user, as the market will not regulate safety or efficacy issues. This paper recommends convergence toward an international validation and surveillance framework based on the specificities of BITs, not equivalent to medical devices, to guarantee their effectiveness and safety for users.


Arthritis and meningitis--the first manifestations of bacterial endocarditis in 2 patients.
We have encountered 2 patients in whom the first manifestations of bacterial endocarditis were arthritis (in 1 case septic arthritis and in the other nonseptic arthritis) and bacterial meningitis. These presentations were followed by acute heart failure due to aortic valve destruction, although the patients showed no significant cardiovascular manifestations on admission. Aortic valve replacement was performed in each case and the patients' postoperative course was comfortable. We would like to emphasize the following points. (1) Arthritis and meningitis are uncommon in patients with bacterial endocarditis. However, it is necessary to consider the possibility of bacterial endocarditis when these clinical manifestations present together. Such a combination can cause rapid valve destruction. When more than 2 rare complications of bacterial endocarditis coexist, surgery should be considered as soon as the definite diagnosis of bacterial endocarditis is established, even if congestive heart failure has not yet developed. (2) Arthritis associated with bacterial endocarditis might be truly septic rather than mediated by circulating immune complexes as is commonly believed.


Modern concepts of diagnosis and treatment of the thoracic outlet syndrome.
Neurovascular compression at the thoracic outlet results from one or several local anatomic abnormalities, all involving the first rib to some extent. The pathologic processes produced involve syndromes usually affecting the ulnar nerve, the subclavian artery, or the subclavian vein. The majority of these cases should be managed in a nonoperative fashion and surgical treatment reserved for patients who are unresponsive to these measures. Initial conservative nonoperative treatment with physiotherapy and patient education produces satisfactory results in 70 per cent of the patients. Operative intervention, when necessary, results in satisfactory results in 90 per cent of the cases and requires first rib resection in conjunction with repair of all other attendant mechanisms. The anterior operative approach is strongly recommended whenever vascular repair is contemplated.


Expressive arts therapy for hospitalized children: a pilot study measuring cortisol levels.
This pilot study aimed at assessing the feasibility of capturing physiological evidence of reduced stress for hospitalized children following expressive arts therapy. Twenty-five patients were offered a novel form of expressive arts therapy, termed Healing Sock Creatures, during their stay in the hospital. Saliva samples were collected at two times in the afternoon for the purpose of measuring salivary cortisol levels. The patients were randomly assigned to two groups, a treatment group or a wait-list control group. A trend of decreased cortisol levels was apparent following therapy in the treatment group and concurrent steroid treatment, which is common in intensive care units, does not appear to interfere with the ability to measure decreased cortisol levels following therapy. Our results support the design of a formal study to assess physiological biomarkers of stress in hospital settings. To our knowledge, this is the first in-patient study assessing a biomarker of stress following expressive arts therapy for children.


Location of the human frontal eye field as defined by electrical cortical stimulation: anatomical, functional and electrophysiological characteristics.
Electrical cortical stimulation of the human frontal gyri and the precentral gyrus has been shown to induce eye movements and it has classically been assumed that these stimulation-induced eye movements result from electrical interference with the human homologue of the monkey frontal eye field (FEF). However, amplitude of electrical current and induced type of eye movement, which are essential for the determination of eye fields in the monkey, have not been investigated systematically in man. We applied electrical cortical stimulation in the lateral frontal cortex in six epileptic patients. Sites whose stimulation resulted in eye movements were determined with respect to gyral and sulcal patterns, Talairach coordinates and neighboring functions as found by electrical cortical stimulation. Based on this approach, a restricted location of the electrically defined FEF is proposed within a larger oculomotor region on the posterior part of the middle frontal gyrus.


In the face of climate change and exhaustive exercise: the physiological response of an important recreational fish species.
Cobia (Rachycentron canadum) support recreational fisheries along the US mid- and south-Atlantic states and have been recently subjected to increased fishing effort, primarily during their spawning season in coastal habitats where increasing temperatures and expanding hypoxic zones are occurring due to climate change. We therefore undertook a study to quantify the physiological abilities of cobia to withstand increases in temperature and hypoxia, including their ability to recover from exhaustive exercise. Respirometry was conducted on cobia from Chesapeake Bay to determine aerobic scope, critical oxygen saturation, ventilation volume and the time to recover from exhaustive exercise under temperature and oxygen conditions projected to be more common in inshore areas by the middle and end of this century. Cobia physiologically tolerated predicted mid- and end-of-century temperatures (28-32°C) and oxygen concentrations as low as 1.7-2.4 mg l-1. Our results indicated cobia can withstand environmental fluctuations that occur in coastal habitats and the broad environmental conditions their prey items can tolerate. However, at these high temperatures, some cobia did suffer post-exercise mortality. It appears cobia will be able to withstand near-future climate impacts in coastal habitats like Chesapeake Bay, but as conditions worsen, catch-and-release fishing may result in higher mortality than under present conditions.


A Si/Glass Bulk-Micromachined Cryogenic Heat Exchanger for High Heat Loads: Fabrication, Test, and Application Results.
This paper reports on a micromachined Si/glass stack recuperative heat exchanger with in situ temperature sensors. Numerous high-conductivity silicon plates with integrated platinum resistance temperature detectors (Pt RTDs) are stacked, alternating with low-conductivity Pyrex spacers. The device has a 1 x 1-cm(2) footprint and a length of up to 3.5 cm. It is intended for use in Joule-Thomson (J-T) coolers and can sustain pressure exceeding 1 MPa. Tests at cold-end inlet temperatures of 237 K-252 K show that the heat exchanger effectiveness is 0.9 with 0.039-g/s helium mass flow rate. The integrated Pt RTDs present a linear response of 0.26%-0.30%/K over an operational range of 205 K-296 K but remain usable at lower temperatures. In self-cooling tests with ethane as the working fluid, a J-T system with the heat exchanger drops 76.1 K below the inlet temperature, achieving 218.7 K for a pressure of 835.8 kPa. The system reaches 200 K in transient state; further cooling is limited by impurities that freeze within the flow stream. In J-T self-cooling tests with an external heat load, the system reaches 239 K while providing 1 W of cooling. In all cases, there is an additional parasitic heat load estimated at 300-500 mW.


Sight-threatening optic neuropathy is associated with paranasal lymphoma.
Malignant lymphoma around the orbit is very rare. We present a rare case of optic neuropathy caused by lymphoma. A 61-year-old Japanese woman was referred to our hospital for evaluation of idiopathic optic neuropathy affecting her right eye. The patient was treated with steroid pulse therapy (methyl-predonisolone 1 g daily for 3 days) with a presumed diagnosis of idiopathic optic neuritis. After she had been switched to oral steroid therapy, endoscopic sinus surgery had been performed, which revealed diffuse large B cell lymphoma of the ethmoidal sinus. Although R-CHOP therapy was immediately started, prolonged optic nerve compression resulted in irreversible blindness. Accordingly, patients with suspected idiopathic optic neuritis should be carefully assessed when they show a poor response, and imaging of the orbits and brain should always be done for initial diagnosis because they may have compression by a tumor.


Salpingo-oophorectomy or surveillance for ovarian endometrioma in asymptomatic premenopausal women: a cost-effectiveness analysis.
The lifetime risk of ovarian cancer is 1.9% among women with endometriosis compared with 1.3% among the general population. When an asymptomatic endometrioma is incidentally discovered on imaging, gynecologists must weigh the procedural complications and the potential for subsequent surgical menopause against future ovarian pathology or cancer. We aimed to determine if performing unilateral salpingo-oophorectomy is a more cost-effective strategy for the prevention of death than surveillance for asymptomatic endometriomas. We created a cost-effectiveness model using TreeAge Pro (TreeAge Software Inc; Williamstown, MA) with a lifetime horizon. Our hypothetical cohort included premenopausal patients with 2 ovaries who did not desire fertility. Those diagnosed with asymptomatic endometrioma underwent either unilateral salpingo-oophorectomy or surveillance (ultrasound 6-12 weeks after diagnosis, then annually). Our primary effectiveness outcome was mortality, including death from ovarian cancer or surgery and all-cause mortality related to surgical menopause (± hormone replacement therapy) if the contralateral ovary is removed. We modeled the probabilities of surgical complications, occult malignancy, development of contralateral adnexal pathology, surgical menopause, use of hormone replacement therapy, and development of ovarian cancer. The costs included surgical procedures, complications, ultrasound surveillance, hormone therapy, and treatment of ovarian cancer, with information gathered from Medicare reimbursement data and published literature. Cost-effectiveness was determined using the incremental cost-effectiveness ratio of Δ costs / Δ deaths with a willingness-to-pay threshold of $11.6 million as the value of a statistical life. Multiple 1-way sensitivity analyses were performed to evaluate model robustness. Our model demonstrated that unilateral salpingo-oophorectomy is associated with improved outcomes compared with surveillance, with fewer deaths (0.28% vs 1.50%) and fewer cases of ovarian cancer (0.42% vs 2.96%). However, it costs more than sonographic surveillance at $6403.43 vs $5381.39 per case of incidental endometrioma. The incremental cost-effectiveness ratio showed that unilateral salpingo-oophorectomy costs $83,773.77 per death prevented and $40,237.80 per case of ovarian cancer prevented. As both values were well below the willingness-to-pay threshold, unilateral salpingo-oophorectomy is cost-effective and is the preferred strategy. If unilateral salpingo-oophorectomy were chosen over surveillance for premenopausal patients with incidental endometriomas, 1 diagnosis of ovarian cancer would be prevented in every 40 patients and 1 death averted in every 82 patients. We performed 1-way sensitivity analyses for all input variables and determined that there were no reasonable inputs that would alter our conclusions. Unilateral salpingo-oophorectomy is cost-effective and is the preferred strategy compared with surveillance for the management of incidental endometrioma in a premenopausal patient not desiring fertility. It incurs fewer deaths and fewer cases of ovarian cancer with costs below the national willingness-to-pay thresholds.


Termination of the peripubertal FSH increase in male rats.
To determine if the pubertal testosterone rise plays a role in the termination of the peripubertal follicle-stimulating hormone (FSH) increase, male rats were injected with ethylene dimethanesulfonate (EDS) at 40 days of age to eradicate the Leydig cells just before the onset of the testosterone rise. Rats were decapitated at weekly intervals from age 26 to 96 days. Compared with vehicle-injected controls, EDS treatment resulted in a delay in the peripubertal increase in the relative weights of prostates and seminal vesicles of approximately 2 wk. Serum testosterone remained at pretreatment levels for 1 wk postinjection. Testicular interstitial fluid testosterone remained at pretreatment concentrations for considerably longer and was significantly lower than controls for 2 wk postinjection. EDS treatment resulted in serum FSH levels that were elevated by 1 wk postinjection. They remained significantly higher than controls until 96 days of age. Compared with controls, serum alpha-inhibin was elevated after EDS as was serum luteinizing hormone. These results suggest that the pubertal testosterone increase plays an important role in terminating the peripubertal FSH rise.


Total force fitness: the military family fitness model.
The military lifestyle can create formidable challenges for military families. This article describes the Military Family Fitness Model (MFFM), a comprehensive model aimed at enhancing family fitness and resilience across the life span. This model is intended for use by Service members, their families, leaders, and health care providers but also has broader applications for all families. The MFFM has three core components: (1) family demands, (2) resources (including individual resources, family resources, and external resources), and (3) family outcomes (including related metrics). The MFFM proposes that resources from the individual, family, and external areas promote fitness, bolster resilience, and foster well-being for the family. The MFFM highlights each resource level for the purpose of improving family fitness and resilience over time. The MFFM both builds on existing family strengths and encourages the development of new family strengths through resource-acquiring behaviors. The purpose of this article is to (1) expand the military's Total Force Fitness (TFF) intent as it relates to families and (2) offer a family fitness model. This article will summarize relevant evidence, provide supportive theory, describe the model, and proffer metrics that support the dimensions of this model.


Microbial reductive debromination of polybrominated diphenyl ethers (PBDEs).
Polybrominated diphenyl ethers (PBDEs) are a class of widely used flame retardants that have recently been detected in environmental samples, diverse biota, human blood serum, and breast milk at exponentially increasing concentrations. Currently, little is known about the fate of these compounds, and in particular, about the microbial potential to degrade them. In this study, debromination of deca-BDE and an octa-BDE mixture is demonstrated with anaerobic bacteria including Sulfurospirillum multivorans and Dehalococcoides species. Hepta- and octa-BDEs were produced by the S. multivorans culture when it was exposed to deca-BDE, although no debromination was observed with the octa-BDE mixture. In contrast, a variety of hepta- through di-BDEs were produced by Dehalococcoides-containing cultures exposed to an octa-BDE mixture, despite the fact that none of these cultures could debrominate deca-BDE. The more toxic hexa-154, penta-99, tetra-49, and tetra-47 were identified among the debromination products. Because the penta-BDE congeners are among the most toxic PBDEs, debromination of the higher congeners to more toxic products in the environment could have profound implications for public health and for the regulation of these compounds.


Impaired glucose-induced insulin response in transgenic mice overexpressing the L-phosphofructokinase gene.
The selective impairment of glucose-induced insulin secretion in NIDDM can be attributed to defects in the glucose-signaling system. An alteration in the activity of phosphofructokinase (PFK), a key enzyme in the glycolytic pathway, may play a role in the abnormal glucose-induced insulin secretion. In this study, we evaluated insulin secretion in transgenic (Tg) mice overexpressing the liver-type subunit of phosphofructokinase (PFKL). Three independently derived Tg-PFKL lines showed random and postprandial hyperglycemia with diminished acute insulin response following intravenous glucose tolerance load. Isolated islets of Tg-PFKL mice exhibited a shift to the right of the glucose insulin dose curve. However, the maximal insulin secretory capacity, as well as the potentiation effect by arginine, were retained. PFK activity in Tg-PFKL islets was increased by 30-70%, because of the overexpression of PFKL. Conceivably, a selective overexpression of the PFKL isoform in Tg-PFKL mice altered the enzymatic properties of the tetrameric PFK and thereby affected glucose metabolism. A similar phenomenon was previously observed in transfected PC12-PFKL cells. The data show that overexpression of PFKL in transgenic mice was associated with diminished glucose-induced insulin response and suggest a mechanism to explain the role of beta-cell PFK activity in glucose-induced insulin secretion.


Effects of vagotomy and drugs on the ventricular fibrillation threshold of normal and ischemic hearts in the anesthetized rat.
The ventricular fibrillation threshold (VFT) was determined by the electrical stimulation of the normal and the coronary artery-ligated myocardium in situ in anesthetized rats. A bilateral vagotomy lowered the VFT of the normal myocardium. After the coronary artery ligation in the vagotomized rats, VFT fell to a greater extent. Bretylium 20-40 mg/kg i.v. produced an elevation of VFT, hypotension and a decrease of the heart rate of rats with the normal or the ischemic myocardium. Lidocaine 10 mg/kg i.v. insignificantly affected the VFT of the normal and the ischemic myocardium. Nitroprusside 4 mg/kg i.v. elevated the VFT of the normal myocardium, and prevented a fall in the VFT of the ischemic myocardium. Adenosine 10 mg/kg or aspirin 20 mg/kg i.v. had no remarkable effect on the VFT of the normal myocardium, but prevented a fall in the VFT of the ischemic myocardium.


FoxO3 mediates antagonistic effects of glucocorticoids and interleukin-2 on glucocorticoid-induced leucine zipper expression.
We have analyzed the promoter of human gilz (glucocorticoid-induced leucine zipper), a dexamethasone-inducible gene that is involved in regulating apoptosis, and identified six glucocorticoid (GC)-responsive elements and three Forkhead responsive elements (FHREs). Promoter deletion analysis and point mutations showed that individual mutation of the GC-responsive elements does not affect GC-induced transcription and that FHRE-1 and FHRE-3 elements contribute to the effects of GCs. Furthermore, overexpression of the Forkhead transcription factor FoxO3 enhances GC-induced gilz mRNA expression. The functional significance of the interaction between FoxO3 and GC receptor was established in T lymphocytes. Indeed, we show that GCs failed to induce GILZ expression in the presence of IL-2, a cytokine known to antagonize GC effects in T cells. Using a constitutive active mutant of protein kinase B that inactivates FoxO3 or a FoxO3 mutant that cannot be inactivated by protein kinase B, we demonstrate that IL-2 inhibitory effects on GILZ expression are mediated through inhibition of FoxO3 transcriptional activity. Therefore, FoxO3 appears to be a key factor mediating GC and IL-2 antagonism for gilz regulation in T lymphocytes. This regulation of GILZ expression was placed in a meaningful context in evaluating the effects of GILZ on GC-induced apoptosis in T lymphocytes.


Amyloid Goiter in a Patient with Rheumatoid Arthritis and End-Stage Renal Disease.
The association between amyloidosis and collagen vascular diseases, such as rheumatoid arthritis (RA) is well-documented. Amyloid goiter is an extremely rare pathologic condition caused by a massive amyloid infiltration of the thyroid tissue. Our patient had been diagnosed with RA 20 years ago and was on hemodialysis for 7 years. He was assessed for decreased appetite, dysphagia, and nausea during the hemodialysis. On physical examination, the thyroid was diffusely enlarged with multiple nodules. He was biochemically euthyroid. Ultrasound of the thyroid gland showed multinodular goiter. A total thyroidectomy was performed. Histopathological examination showed dilated follicles surrounded by abundant homogeneous substance that stained positive with Congo red. The patient was reported as amyloid goiter. Complaints of the patient improved after the surgery. In the literature, amyloid goiter with RA in a hemodialysis patient is very rare. Amyloid goiter should be considered if there is a rapid thyromegaly causing pressure symptoms in the background of any disease with chronic inflammation.


Automated data reduction for hydrogen/deuterium exchange experiments, enabled by high-resolution Fourier transform ion cyclotron resonance mass spectrometry.
Mass analysis of proteolytic fragment peptides following hydrogen/deuterium exchange offers a general measure of solvent accessibility/hydrogen bonding (and thus conformation) of solution-phase proteins and their complexes. The primary problem in such mass analyses is reliable and rapid assignment of mass spectral peaks to the correct charge state and degree of deuteration of each fragment peptide, in the presence of substantial overlap between isotopic distributions of target peptides, autolysis products, and other interferant species. Here, we show that at sufficiently high mass resolving power (m/Delta m(50%) > or = 100,000), it becomes possible to resolve enough of those overlaps so that automated data reduction becomes possible, based on the actual elemental composition of each peptide without the need to deconvolve isotopic distributions. We demonstrate automated, rapid, reliable assignment of peptide masses from H/D exchange experiments, based on electrospray ionization FT-ICR mass spectra from H/D exchange of solution-phase myoglobin. Combined with previously demonstrated automated data acquisition for such experiments, the present data reduction algorithm enhances automation (and thus expands generality and applicability) for high-resolution mass spectrometry-based analysis of H/D exchange of solution-phase proteins.


Hearing impairment in murine model of Down syndrome.
Hearing impairment is a cardinal feature of Down syndrome (DS), but its clinical manifestations have been attributed to multiple factors. Murine models could provide mechanistic insights on various causes of hearing loss in DS. To investigate mechanisms of hearing loss in DS in the absence of the cadherin 23 mutation, we backcrossed our DS mice, Dp(16)1Yey, onto normal-hearing CBA/J mice and evaluated their auditory function. Body weights of wild type (WT) and DS mice were similar at 3-months of age, but at 9-months, WT weighed 30% more than DS mice. Distortion product otoacoustic emissions (DPOAE), a test of sensory outer hair cell (OHC) function negatively impacted by conductive hearing loss, were reduced in amplitude and sensitivity across all frequencies in DS mice. The middle ear space in DS mice appeared normal with no evidence of infection. MicroCT structural imaging of DS temporal bones revealed a smaller tympanic membrane diameter, oval window, and middle ear space and localized thickening of the bony otic capsule, but no gross abnormalities of the middle ear ossicles. Histological analysis of the cochlear and vestibular sensory epithelium revealed a normal density of cochlear and vestibular hair cells; however, the cochlear basal membrane was approximately 0.6 mm shorter in DS than WT mice so that the total number of hair cells was greater in WT than DS mice. In DS mice, the early and late peaks in the auditory brainstem response (ABR), reflecting neural responses from the cochlear auditory nerve followed by subsequent neural centers in the brainstem, were reduced in amplitude and ABR thresholds were elevated to a similar degree across all frequencies, consistent with a conductive hearing impairment. The latency of the peaks in the ABR waveform were longer in DS than WT mice when compared at the same intensity; however, the latency delays disappeared when the data were compared at the same intensity above thresholds to compensate for the conductive hearing loss. Future studies using wideband tympanometry and absorbance together with detailed histological analysis of the middle ear could illuminate the nature of the conductive hearing impairment in DS mice.


Biological effects of simple changes in functionality on rhodium metalloinsertors.
DNA mismatch repair (MMR) is crucial to ensuring the fidelity of the genome. The inability to correct single base mismatches leads to elevated mutation rates and carcinogenesis. Using metalloinsertors-bulky metal complexes that bind with high specificity to mismatched sites in the DNA duplex-our laboratory has adopted a new chemotherapeutic strategy through the selective targeting of MMR-deficient cells, that is, those that have a propensity for cancerous transformation. Rhodium metalloinsertors display inhibitory effects selectively in cells that are deficient in the MMR machinery, consistent with this strategy. However, a highly sensitive structure-function relationship is emerging with the development of new complexes that highlights the importance of subcellular localization. We have found that small structural modifications, for example a hydroxyl versus a methyl functional group, can yield profound differences in biological function. Despite similar binding affinities and selectivities for DNA mismatches, only one metalloinsertor shows selective inhibition of cellular proliferation in MMR-deficient versus -proficient cells. Studies of whole-cell, nuclear and mitochondrial uptake reveal that this selectivity depends upon targeting DNA mismatches in the cell nucleus.


A Comprehensive Rehabilitation Program and Follow-up Assessment for Acute Intermittent Porphyria.
Acute intermittent porphyria (AIP) is an infrequent metabolic disease that can cause severe disability or death without timely treatment. A porphyric attack occurs when genetic factors combine with trigger factors, and diagnosis may be delayed owing to nonspecific symptoms. Recovery from AIP can be nearly or fully complete with proper treatment, which includes intravenous hematin administration, the control of trigger factors, and a comprehensive rehabilitation program. The aim of this case report was to describe the clinical evolution of a 43-year-old woman with AIP and a polyneuropathy. The patient was treated through a comprehensive rehabilitation program, with outcomes evaluated by the Functional Independence Measure and the Berg scales during rehabilitation and postdischarge follow-up. After completing the comprehensive rehabilitation program, the patient achieved a satisfactory level of functional independence, allowing for social and work reintegration. We conclude that an early and multidisciplinary approach is essential for regaining optimal functionality after AIP.


One-two punch injury to tolerance mechanisms in graft-versus-host disease.
Chronic graft-versus-host disease (GVHD) is a major complication of allogeneic hematopoietic cell transplantation that resembles autoimmunity, with unclear pathogenesis and few effective therapeutic options. In this issue of the JCI, Dertschnig et al. used mouse models to investigate the basis of T cell autoreactivity following GVHD. Notably, GVHD caused irreversible damage to a population of tolerogenic stromal cells that display peripheral tissue-restricted antigens in lymph nodes, which impaired their capacity to purge and suppress autoreactive T cells. Together with damage to central tolerance mechanisms in the thymus, these findings outline a critical one-two punch injury that profoundly disrupts immune tolerance in this devastating disease.


Understanding print: early reading development and the contributions of home literacy experiences.
This study explored the development of children's early understanding of visual and orthographic aspects of print and how this is related to early reading acquisition. A total of 474 children, ages 48 to 83 months, completed standardized measures of phonological awareness and early reading skills. They also completed experimental tasks that tapped their understanding of what constitutes "readable" print. The parents of participants completed a questionnaire regarding their children's home literacy experiences. The data showed systematic development in children's understanding of print conventions and English orthography and spelling. Regression analyses indicated that print knowledge was related to early reading skill, even after accounting for variance due to age and phonological awareness. Furthermore, parents' ratings of the extent of their children's involvement in activities that led to practice in reading and writing most consistently predicted the development of emerging literacy skills, including understanding of the conventions of the English writing system. Little relation between print knowledge and the frequency of storybook reading by adults was observed.


PAs and NPs in HMOs.
Recognition of NPP contributions to the delivery of care in HMOs has come slowly. Staffing continues to be a function of physician attitude instead of management rationale. Only a few enlightened HMOs seek management and staffing input from NPPs. Investigation of the role of NPPs in HMOs has seriously lagged behind that of physicians, nurses, pharmacists, and other health professionals. The status, reward systems, and job satisfaction of nonphysician clinicians, as well as efforts and problems in recruiting them, have been inadequately reported. Only a handful of HMOs have contributed to our understanding of NPP productivity and task delegation. If HMOs plan to continue using NPPs, much more information on optimizing their deployment must be shared. Member satisfaction surveys must be reported, prescribing practices must be compared, liability issues explored, and outcomes research pursued. We know little about the outcome of nonacute illness managed by NPPs. Do NPPs negate their cost-effectiveness by using more or different resources? Do differences between PAs and NPs exist with regard to each of these issues? Because so many study variables can be held constant by a large HMO which provides care in a given area under uniform market forces, HMOs can best explore how to maximize NPP effectiveness.


Mechanisms controlling cell size and shape during isotropic cell spreading.
Cell motility is important for many developmental and physiological processes. Motility arises from interactions between physical forces at the cell surface membrane and the biochemical reactions that control the actin cytoskeleton. To computationally analyze how these factors interact, we built a three-dimensional stochastic model of the experimentally observed isotropic spreading phase of mammalian fibroblasts. The multiscale model is composed at the microscopic levels of three actin filament remodeling reactions that occur stochastically in space and time, and these reactions are regulated by the membrane forces due to membrane surface resistance (load) and bending energy. The macroscopic output of the model (isotropic spreading of the whole cell) occurs due to the movement of the leading edge, resulting solely from membrane force-constrained biochemical reactions. Numerical simulations indicate that our model qualitatively captures the experimentally observed isotropic cell-spreading behavior. The model predicts that increasing the capping protein concentration will lead to a proportional decrease in the spread radius of the cell. This prediction was experimentally confirmed with the use of Cytochalasin D, which caps growing actin filaments. Similarly, the predicted effect of actin monomer concentration was experimentally verified by using Latrunculin A. Parameter variation analyses indicate that membrane physical forces control cell shape during spreading, whereas the biochemical reactions underlying actin cytoskeleton dynamics control cell size (i.e., the rate of spreading). Thus, during cell spreading, a balance between the biochemical and biophysical properties determines the cell size and shape. These mechanistic insights can provide a format for understanding how force and chemical signals together modulate cellular regulatory networks to control cell motility.


Relation between cytosolic free Ca2+ concentration and the control of pyruvate dehydrogenase in isolated cardiac myocytes.
The proportion of pyruvate dehydrogenase existing in the active form (PDHA) in suspensions of unstimulated cardiac myocytes oxidizing glucose is approx. 30%. Depolarization of the cells with concentrations of K+ above physiological values leads to an increase in the content of PDHA. Overloading of the cells with Na+ by treatment with veratridine and ouabain gives the same result. Each of these interventions is shown in experiments with Quin 2-loaded myocytes to lead to an increase in cytosolic free Ca2+ concentration ([Ca2+]c). Treatment of the cells with Ruthenium Red, an inhibitor of Ca2+ transport into mitochondria, largely prevents an increase in PDHA in response to addition of KCl or of veratridine plus ouabain. Ruthenium Red does not attenuate the increase in [Ca2+]c that occurs under these conditions. By contrast, treatment of the cells with ryanodine, an inhibitor of sarcoplasmic-reticulum Ca2+ transport and therefore of contraction, does not diminish the response of PDHA content to agents which raise [Ca2+]c; nor does loading of the cells with the Ca2+-chelating agent Quin 2, which also prevents contraction, at appropriate concentrations. It is concluded that an increase in [Ca2+]c causes an increase in PDHA content of cardiac myocytes independently of an increase in mechanical work. In the normal physiological situation the activation of dehydrogenases by Ca2+ is thought to help to maintain the balance of energy supply and demand during periods of increased work-load, which are associated with an increased myoplasmic [Ca2+]c.


Balancing of mitochondrial translation through METTL8-mediated m3C modification of mitochondrial tRNAs.
Mitochondria contain a specific translation machinery for the synthesis of mitochondria-encoded respiratory chain components. Mitochondrial tRNAs (mt-tRNAs) are also generated from the mitochondrial DNA and, similar to their cytoplasmic counterparts, are post-transcriptionally modified. Here, we find that the RNA methyltransferase METTL8 is a mitochondrial protein that facilitates 3-methyl-cytidine (m3C) methylation at position C32 of the mt-tRNASer(UCN) and mt-tRNAThr. METTL8 knockout cells show a reduction in respiratory chain activity, whereas overexpression increases activity. In pancreatic cancer, METTL8 levels are high, which correlates with lower patient survival and an enhanced respiratory chain activity. Mitochondrial ribosome profiling uncovered mitoribosome stalling on mt-tRNASer(UCN)- and mt-tRNAThr-dependent codons. Further analysis of the respiratory chain complexes using mass spectrometry revealed reduced incorporation of the mitochondrially encoded proteins ND6 and ND1 into complex I. The well-balanced translation of mt-tRNASer(UCN)- and mt-tRNAThr-dependent codons through METTL8-mediated m3C32 methylation might, therefore, facilitate the optimal composition and function of the mitochondrial respiratory chain.


ABC50 interacts with eukaryotic initiation factor 2 and associates with the ribosome in an ATP-dependent manner.
Eukaryotic initiation factor 2 (eIF2) plays a key role in the process of translation initiation and in its control. Here we demonstrate that highly purified mammalian eIF2 contains an additional polypeptide of apparent molecular mass of 110 kDa. This polypeptide co-purified with eIF2 through five different chromatography procedures. A cDNA clone encoding the polypeptide was isolated, and its sequence closely matched that of a protein previously termed ABC50, a member of the ATP-binding cassette (ABC) family of proteins. Antibodies to ABC50 co-immunoprecipitated eIF2 and vice versa, indicating that the two proteins interact. The presence of ABC50 had no effect upon the ability of eIF2 to bind GDP but markedly enhanced the association of methionyl-tRNA with the factor. Unlike the majority of ABC proteins, which are membrane-associated transporters, ABC50 associates with the ribosome and co-sediments in sucrose gradients with the 40 and 60 S ribosomal subunits. The association of ABC50 with ribosomal subunits was increased by ATP and decreased by ADP. ABC50 is related to GCN20 and eEF3, two yeast ABC proteins that are not membrane-associated transporters and are instead implicated in mRNA translation and/or its control. Thus, these data identify ABC50 as a third ABC protein with a likely function in mRNA translation, which associates with eIF2 and with ribosomes.


Retinal artery occlusion followed by contralateral amaurosis fugax in association with eosinophilic granulomatosis with polyangiitis (Churg-Strauss syndrome).
To describe two cases of retinal artery occlusion followed by contralateral amaurosis fugax associated with eosinophilic granulomatosis with polyangiitis (EGPA, formerly known as Churg-Strauss syndrome). Case 1 is a 57 year-old male who presented with transient vision loss in the right eye two weeks after a cilioretinal artery occlusion in the left eye. Evaluation eventually led to a diagnosis of EGPA. The patient was treated with high-dose steroids followed by systemic immunomodulatory therapy. Vision in the right eye recovered to 20/20 with no further episodes of vision loss. Case 2 is a 55 year-old male with a known diagnosis of EGPA who presented with transient vision loss in the right eye four weeks after a central retinal artery occlusion of the left eye. This patient also successfully recovered vision in the right eye after treatment with high-dose steroids following a change in his systemic immunomodulatory therapy. While ANCA-vasculitides are an uncommon cause of retinal artery occlusion and amaurosis fugax, it is important that they remain in the differential diagnosis, as good visual outcomes can be achieved with prompt initiation of appropriate therapies.