Cross-situational statistically based word learning intervention for late-talking toddlers.
To explore the efficacy of a word learning intervention for late-talking toddlers that is based on principles of cross-situational statistical learning. Four late-talking toddlers were individually provided with 7-10 weeks of bi-weekly word learning intervention that incorporated principles of cross-situational statistical learning. Treatment was input-based meaning that, aside from initial probes, children were not asked to produce any language during the sessions. Pre-intervention data included parent-reported measures of productive vocabulary and language samples. Data collected during intervention included production on probes, spontaneous production during treatment, and parent report of words used spontaneously at home. Data were analyzed for number of target words learned relative to control words, effect sizes, and pre-post treatment vocabulary measures. All children learned more target words than control words and, on average, showed a large treatment effect size. Children made pre-post vocabulary gains, increasing their percentile scores on the MCDI, and demonstrated a rate of word learning that was faster than rates found in the literature. Cross-situational statistically based word learning intervention has the potential to improve vocabulary learning in late-talking toddlers. Limitations on interpretation are also discussed. Readers will describe what cross-situational learning is and how it might apply to treatment. They will identify how including lexical and contextual variability in a word learning intervention for toddlers affected treatment outcomes. They will also recognize evidence of improved rate of vocabulary learning following treatment.


The role of environmental stress on lower urinary tract symptoms.
Lower urinary tract symptoms (LUTS) have been associated with comorbid conditions such as anxiety and depression. In addition, stress appears to influence the development or exacerbation of LUTS. This article seeks to review literature regarding the role of environmental stress on LUTS, focusing on findings presented in the last year. Numerous authors have published on the impact early childhood experiences, acute and chronic stress, and psychiatric illness play in the development of LUTS. The exact nature of the association between bladder symptoms and psychosocial measures remains unknown and is likely due to a complex interplay between heritability, psychosocial factors, and environmental stress. The proposed pathophysiological pathways involved in emotional states such as anxiety and depression, stress, and bladder function include activation of the hypothalamic-pituitary axis, dysregulation of the serotonergic pathways, and central sensitization. Recent work has additionally suggested that urinary syndromes involving abnormal or augmented sensory input, such as overactive bladder and interstitial cystitis/bladder pain syndrome, may be a spectrum of the same disorder. There are numerous developments in our understanding of the role of environmental stress on the development and exacerbation of LUTS with new developments both clinically and in translational basic science work. Clinicians must acknowledge the high prevalence of affective disorders in patients with LUTS and realize their potential therapeutic influence. Simply addressing mechanisms at the level of the bladder alone may fail in a subpopulation of patients with LUTS who may have significant psychosocial drivers of their symptoms.


Membrane structure and surface coat of Entamoeba histolytica. Topochemistry and dynamics of the cell surface: cap formation and microexudate.
Treatment of living entamoeba histolytica cells with low concentrations of concanavalin A (con A) and peroxidase results in redistribution of the plasma membrane con A receptors to one pole of the cell where a morphologically distinct region--the uroid--is formed. Capping of con A receptors is not accompanied by parallel accumulation of ruthenium red-stainable components. In capped cells, the pattern of distribution of acidic sites ionized at pH 1.8 (labeled by colloidal iron) at the outer surface and of membrane particles (integral membrane components revealed by freeze-fracture) is not altered over the uroid region. Cytochemistry of substrate-attached microexudate located in regions adjacent to E. histolytica cells demonstrates the presence of con A binding sites and ruthenium red- and alcian blue-stainable components and the absent of colloidal iron binding sites. In a previous report we demonstrated that glycerol-induced aggregation of the plasma membrane particles is accompanied by a discontinuous distribution of colloidal iron binding sites, while con A receptors and acidic sites ionized at pH 4.0 remain uniformly distributed over the cell surface. Taken together, our experiments show that, in E. histolytica cells, peripheral membrane components may move independently of integral components and, also, that certain surface determinants may redistribute independently of others. These results point to the complexity of the membrane structure-cell surface relationship in E. histolytica plasma membranes relative to the membrane of the erythrocyte ghost where integral components (the membrane-intercalated particles) contain all antigens, receptors, and anionic sites labeled so far. We conclude that fluidity of integral membrane components (integral membrane fluidity) cannot be inferred from the demonstration of the mobility of surface components nor, conversely, can the fluidity of peripheral membrane components (peripheral membrane fluidity) be assumed from demonstration of the mobility of integral membrane components.


Genetics and public health--evolution, or revolution?
During the 19th and early 20th century, public health and genetics shared common ground through similar approaches to health promotion in the population. By the mid-20th century there was a division between public health and genetics, with eugenicists estranged and clinical genetics focused on single gene disorders, usually only relevant to small numbers of people. Now through a common interest in the aetiology of complex diseases such as heart disease and cancer, there is a need for people working in public health and genetics to collaborate. This is not a comfortable convergence for many, particularly those in public health. Nine main concerns are reviewed: fear of eugenics; genetic reductionism; predictive power of genes; non-modifiable risk factors; rights of individuals compared with populations; resource allocation; commercial imperative; discrimination; and understanding and education. This paper aims to contribute to the thinking and discussion about an evolutionary, multidisciplinary approach to understanding, preventing, and treating complex diseases.


Music Interventions in Pediatric Surgery (The Music Under Surgery In Children Study): A Randomized Clinical Trial.
Perioperative music interventions have been shown to reduce anxiety and pain in adults. This inexpensive, easily applicable intervention could be of benefit to children as well. Our objective was to determine the effects of music interventions on distress, anxiety, and postoperative pain in infants undergoing surgery. The Music Under Surgery In Children study was designed as a parallel, single-blind, randomized controlled trial with an a priori formulated hypothesis. Data were collected between August 2015 and October 2016 in a single tertiary care children's hospital. There was a 24-hour follow-up with blind primary outcome assessment. A random sample of 432 eligible 0-3 years of age infants admitted for orchidopexy, hypospadias, or inguinal hernia repair receiving general anesthesia and caudal block were asked for participation. Subjects were assigned to a preoperative music intervention, pre- and intraoperative music intervention, or no music intervention (control) via random allocation using a computer-generated list with the use of opaque envelopes. The main outcome measure was the postoperative level of distress assessed with the COMFORT-Behavior scale, which is an observational scale; furthermore, preoperative level of distress, preoperative anxiety, and physiological measurements such as heart rate (HR) and blood pressure were measured. The trial was registered at the Dutch Trial Register, number NTR5402 (www.trialregister.nl). One hundred ninety-five infants with median age 6.9 months (interquartile range, 3.3-11.1) were randomized, 178 of whom were included in the primary analysis. A nonsignificant difference in COMFORT-Behavior scale scores between the pre- and intraoperative music intervention group and control group at 4 hours after surgery was found (mean difference, -1.22; 95% CI, 2.60-0.17; P = .085). Additional analysis showed weak nonsignificant evidence for an interaction effect between music exposure and COMFORT-Behavior score at baseline (P = .027 with a Bonferroni-adjusted significance level of .025). General linear modeling showed a statistically significantly reduced HR after the preoperative music intervention in the holding area in the combined preoperative music intervention and intraoperative music intervention group compared to the control group (P = .003). The differences in HR among the 3 study arms at all time points were not statistically significant (P = .069). Music interventions do not seem to benefit all young infants undergoing surgery. The potential benefits of music interventions in the preoperative period and in more distressed children warrant further exploration.


[Intracellular recording of potentials of cells of isolated cerebellum in the frog].
Intracellular potentials of granular, Purkinje and glial cells were recorded in an isolated frog cerebellum in response to electrical stimulation of cerebellar peduncle. The stimuli inhibited strongly the activity of most granular cells. "Slow" (20-50 ms) and "fast" (1-2 ms) spikes were recorded in Purkinje cells. Glial cell responses consisted of a slow waves of depolarization summated with repetitive stimuli and reached an amplitude of 20 mV.


Ultrastructural and electrophysiological changes associated with K(+)-evoked release of neurotransmitter at the synaptic terminals of skate photoreceptors.
Bathing the skate retina in a Ringer solution containing a high concentration (100 mM) of potassium ions depolarized the visual cells, depleted the receptor terminals of synaptic vesicles, and suppressed completely the b-wave of the ERG and the intracellularly recorded response of horizontal cells (the S-potential). The depletion of synaptic vesicles was accompanied by a large increase in the extent of the plasma membrane resulting in distortion of the normal terminal profile, i.e. distension of the basal surface and elaborate infolding of protoplasmic extensions. Morphometric analysis showed that despite the changes in vesicle content and terminal structure, the combined linear extent of the vesicular and plasma membranes was unchanged from control (superfusion with normal Ringer solution); the increase in plasma membrane was equivalent to the observed loss of vesicular membrane. When returned to a normal Ringer solution, the terminals rapidly began to reform, and in about 10 min they were morphologically indistinguishable from receptor terminals seen in control preparations. After 30 min in the normal Ringer solution, the amount of membrane associated with the vesicles and the plasma membrane had reverted to control values, and once again the total membrane estimated morphometrically remained essentially the same. Thus, there is an efficient mechanism at the photoreceptor terminal for the recycling of vesicle membrane following exocytosis. The K(+)-induced depletion of synaptic vesicles was paralleled by a precipitous loss of responsivity in both the b-wave of the ERG and the S-potential of the horizontal cells. However, after 30-min exposure to the high K+ and a return to normal Ringer solution, the recovery of electrophysiological activity followed a much slower time course from that associated with the structural changes; 60 min or longer were required for the potentials to exhibit maximum response amplitudes. It appears that the rate-limiting step in restoring normal synaptic function following massive depletion of vesicular stores is transmitter resynthesis and vesicle loading rather than vesicle recycling.


Understanding the Meaning of Recovery to Patients Undergoing Abdominal Surgery.
Postoperative recovery is difficult to define or measure. Research addressing interventions aimed to improve recovery after abdominal surgery often focuses on measures such as duration of hospital stay and complication rates. Although these clinical parameters are relevant, understanding patients' perspectives regarding postoperative recovery is fundamental to guiding patient-centered care. To elucidate the meaning of recovery from the perspective of patients undergoing abdominal surgery. This international qualitative study involved semistructured interviews with patients recovering from abdominal surgery from October 2016 to November 2018 in tertiary hospitals in 4 countries (Canada, Italy, Brazil, and Japan). A purposive maximal variation sampling method was used to ensure the recruitment of patients with varying demographic, clinical, and surgical characteristics. Data on race were not collected. Each interview lasted between 1 and 2 hours. Interviews were recorded and then transcribed verbatim. Transcripts were then analyzed using an inductive thematic analysis approach. Data analysis was conducted from July 2019 to September 2019. The qualitative analysis revealed themes reflecting the meaning of recovery from the perspective of patients undergoing abdominal surgery. Thirty patients recovering from abdominal surgery were interviewed (15 [50%] female; mean [SD] age, 57 [18] years; 10 [33%] underwent major surgery; 16 [53%] underwent laparoscopic surgery). The interviews revealed that for patients undergoing abdominal surgery, the meaning of recovery embodied 5 overarching themes: (1) returning to habits and routines, (2) resolution of symptoms, (3) overcoming mental strains, (4) regaining independence, and (5) enjoying life. Themes associating the meaning of recovery to traditional parameters, such as earlier hospital discharge or absence of complications, were not identified in the interviews. This qualitative study suggests that the meaning of recovery from the perspective of patients undergoing abdominal surgery goes beyond traditional clinical parameters. The elements of recovery identified in this study should be taken into account in patient-surgeon discussions about recovery and when developing patient-centered strategies to improve postoperative outcomes.


Breed susceptibility for common surgically treated orthopaedic diseases in 12 dog breeds.
A retrospective case-control study was conducted to estimate breed predisposition for common orthopaedic conditions in 12 popular dog breeds in Norway and Sweden. Orthopaedic conditions investigated were elbow dysplasia (ED); cranial cruciate ligament disease (CCLD); medial patellar luxation (MPL); and fractures of the radius and ulna. Dogs surgically treated for the conditions above at the Swedish and Norwegian University Animal Hospitals between the years 2011 and 2015 were compared with a geographically adjusted control group calculated from the national ID-registries. Logistic regression analyses (stratified for clinic and combined) were used to calculate odds ratios (OR) and 95% confidence intervals. Mixed breed dogs were used as reference. Breeds found at-risk for ED were the Labrador retriever (OR = 5.73), the Rottweiler (OR = 5.63), the German shepherd dog (OR = 3.31) and the Staffordshire bull terrier (OR = 3.08). The Chihuahua was the only breed where an increased risk for MPL (OR = 2.80) was identified. While the Rottweiler was the only breed predisposed for CCLD (OR = 3.96), the results were conflicting for the Labrador retriever (OR = 0.44 in Sweden, 2.85 in Norway); the overall risk was identical to mixed-breed dogs. Most results are in concordance with earlier studies. However, an increased risk of CCLD was not identified for the Labrador retriever, the Staffordshire bull terrier was found to have an increased risk of ED and some country-specific differences were noted. These results highlight the importance of utilising large caseloads and appropriate control groups when breed susceptibility is reported.


Persistence of altered movement patterns during a sit-to-stand task 1 year following unilateral total knee arthroplasty.
Following total knee arthroplasty (TKA), quadriceps femoris muscle strength (force-generating capacity) and functional test scores improve but continue to be lower than those in people without injury. Analysis of the sit-to-stand (STS) task demonstrated side-to-side differences in subjects with TKA, as well as differences between subjects with TKA and control subjects. It was hypothesized that, when using a self-selected starting position, subjects 1 year following TKA would show improvements in strength and movement patterns but would continue to show asymmetries of angles and moments at the hips and knees. Twenty-four subjects (12 subjects with unilateral TKA and 12 control subjects) were recruited; those with TKA were tested 3 months and 1 year following surgery. Motion analysis of an STS task was synchronized with 2 force platforms and electromyography. Outcome measures included joint angles and moments, electromyography, vertical ground reaction forces, muscle strength, and functional performance tests. Subjects with TKA showed improvements in symmetry of motion, strength, and functional performance from 3 months to 1 year following TKA. Compared with control subjects, subjects with TKA relied on increased hip flexion and a larger hip extensor moment to perform the STS task. The increased hip extensor moment demonstrated that subjects adopted a strategy to avoid the use of the quadriceps femoris muscle, yet this strategy persisted as quadriceps femoris muscle strength improved. This pattern may be a learned movement pattern that may not resolve without retraining.


The enzymatic status of circulating lymphocytes as an index of the regenerative process in the lungs under stimulation with pyrimidine and purine derivatives. A clinical experimental study.
Enzyme activities in the circulating lymphocytes permit to judge the character of the regenerative process in the lung, and the severity of the patient's condition; it helps in estimating the prognosis and in evaluating the efficacy of the applied therapy. Potassium orotate and Riboxin intensify the regenerative process in the lung undergoing a compensatory-hypertrophic rearrangement, and enhance the functional activity of circulating lymphocytes as participants of the regenerative process and stimulate the oxidation-reduction processes in lymphocytes.


Multi-layered regulation of intestinal antimicrobial defense.
The gastrointestinal tract of mammals is heavily colonized with a complex and dynamic microbial community. To cope with this complex microbial challenge, multiple epithelial lineages, such as enterocytes and Paneth cells, elaborate a diverse repertoire of protein antibiotics. The gut antimicrobial arsenal encompasses multiple protein families, including defensins, cathelicidins, and C-type lectins. These antimicrobial peptides and proteins play a key role in protecting the host against pathogen challenge, and likely also function to limit invasion of indigenous microbes. It is becoming increasingly apparent that expression of mucosal antimicrobial defenses is tightly controlled. This occurs at multiple levels, including transcriptional regulation in response to bacterial cues, post-translational proteolytic processing, and bacterial regulation of Paneth cell degranulation. Impaired antimicrobial peptide expression has also been implicated in inflammatory bowel disease, underscoring the essential role of antimicrobial defenses in maintaining intestinal homeostasis.


The Origin and Early Radiation of Archosauriforms: Integrating the Skeletal and Footprint Record.
We present a holistic approach to the study of early archosauriform evolution by integrating body and track records. The ichnological record supports a Late Permian-Early Triassic radiation of archosauriforms not well documented by skeletal material, and new footprints from the Upper Permian of the southern Alps (Italy) provide evidence for a diversity not yet sampled by body fossils. The integrative study of body fossil and footprint data supports the hypothesis that archosauriforms had already undergone substantial taxonomic diversification by the Late Permian and that by the Early Triassic archosauromorphs attained a broad geographical distribution over most parts of Pangea. Analysis of body size, as deduced from track size, suggests that archosauriform average body size did not change significantly from the Late Permian to the Early Triassic. A survey of facies yielding both skeletal and track record indicate an ecological preference for inland fluvial (lacustrine) environments for early archosauromorphs. Finally, although more data is needed, Late Permian chirotheriid imprints suggest a shift from sprawling to erect posture in archosauriforms before the end-Permian mass extinction event. We highlight the importance of approaching palaeobiological questions by using all available sources of data, specifically through integrating the body and track fossil record.


Novel role for a Saccharomyces cerevisiae nucleoporin, Nup170p, in chromosome segregation.
We determined that a mutation in the nucleoporin gene NUP170 leads to defects in chromosome transmission fidelity (ctf) and kinetochore integrity in Saccharomyces cerevisiae. A ctf mutant strain, termed s141, shows a transcription readthrough phenotype and stabilizes a dicentric chromosome fragment in two assays for kinetochore integrity. Previously, these assays led to the identification of two essential kinetochore components, Ctf13p and Ctf14p. Thus, s141 represents another ctf mutant involved in the maintenance of kinetochore integrity. We cloned and mapped the gene complementing the ctf mutation of s141 and showed that it is identical to the S. cerevisiae NUP170 gene. A deletion strain of NUP170 (nup170 Delta::HIS3) has a Ctf(-) phenotype similar to the s141 mutant (nup170-141) and also exhibits a kinetochore integrity defect. We identified a second nucleoporin, NUP157, a homologue of NUP170, as a suppressor of the Ctf(-) phenotype of nup170-141 and nup170 Delta::HIS3 strains. However, a deletion of NUP157 or several other nucleoporins did not affect chromosome segregation. Our data suggest that NUP170 encodes a specialized nucleoporin with a unique role in chromosome segregation and possibly kinetochore function.


The ultrastructural effects of prednisolone on the mesenchyme of the palatal shelf in the mouse.
The effect of methylprednisolone on the structure of the mesenchymal component of the palatal shelves of fetal mice was studied at the time of shelf reorientation. Pregnant mice received intraperitoneal injections of 8 mg of prednisolone on days 11.5, 12.5, and 13.5 of pregnancy. The mice were killed on day 14.5, and the specimens were either stained with periodic acid-Schiff (PAS) reagent for examination in the light microscope or processed for examination in the electron microscope. It was found that prednisolone treatment resulted in a reduction in the amount of tannic-acid-staining material in the extracellular spaces. It also resulted in a reduction in the collagen within the shelf. The development of skeletal and smooth muscle cells, neurons, and satellite cells was also delayed and the occurrence within the shelves of cells with a high glycogen content was reduced.


Familial nephrotic syndrome with focal glomerular sclerosis.
Two brothers with nephrotic syndrome are reported. Their parents were first cousins. The nephrotic syndrome developed at the age of 14 and 15 years and was resistant to the treatment with corticosteroid. The elder patient fell into renal failure about 2-1/2 years after the development of nephrotic syndrome, and the younger has demonstrated a progressive decrease in renal function. The renal histopathology in one patient revealed findings typical of focal glomerular sclerosis. In light of the clinical similarities in these two patients, it is likely that they have a familial form of focal glomerular sclerosis.


Clostridium difficile-associated diarrhea: risk factors, diagnostic methods, and treatment.
Clostridium difficile-associated diarrhea (CDAD) has become the most common cause of infectious diarrhea acquired in the hospital, with an estimated 3 million annual cases and an annual cost of $1 billion. Risk factors for CDAD include antibiotic use (especially ampicillin, clindamycin, and cephalosporins), advanced age, and gastrointestinal surgery. Specific diagnosis of CDAD is made with an enzyme immunoassay to detect toxins A and B. Metronidazole remains the initial treatment of choice, with a 95% success rate. Vancomycin is reserved for failures. Despite the high initial success rates, recurrence of CDAD remains a significant problem in 20% to 30% of cases, with increased cost and substantial morbidity. Efforts to prevent CDAD will need to be strengthened, including education and better compliance with isolation, use of gloves, and hand washing.


VISTA.COMP - an engineered checkpoint receptor agonist that potently suppresses T cell-mediated immune responses.
V-domain immunoglobulin suppressor of T cell activation (VISTA) is a recently discovered immune checkpoint ligand that functions to suppress T cell activity. The therapeutic potential of activating this immune checkpoint pathway to reduce inflammatory responses remains untapped, largely due to the inability to derive agonists targeting its unknown receptor. A dimeric construct of the IgV domain of VISTA (VISTA-Fc) was shown to suppress the activation of T cells in vitro. However, this effect required its immobilization on a solid surface, suggesting that VISTA-Fc may display limited efficacy as a VISTA-receptor agonist in vivo. Herein, we have designed a stable pentameric VISTA construct (VISTA.COMP) by genetically fusing its IgV domain to the pentamerization domain from the cartilage oligomeric matrix protein (COMP). In contrast to VISTA-Fc, VISTA.COMP does not require immobilization to inhibit the proliferation of CD4+ T cells undergoing polyclonal activation. Furthermore, we show that VISTA.COMP, but not VISTA-Fc, functions as an immunosuppressive agonist in vivo capable of prolonging the survival of skin allografts in a mouse transplant model as well as rescuing mice from acute concanavalin-A-induced hepatitis. Collectively, we believe our data demonstrate that VISTA.COMP is a checkpoint receptor agonist and the first agent to our knowledge targeting the putative VISTA-receptor to suppress T cell-mediated immune responses.


Inhibition of growth of transformed cells and tumors by an endogenous acceptor of galactosyltransferase.
A galactosyltransferase glycopeptide acceptor purified from human malignant effusions was tested for its effects on cell growth in vitro and in vivo. Addition of the glycopeptide to the media of cells growing in tissue culture caused a significant inhibition of attachment and growth of transformed cells but had minimal effect on nontransformed cells. Transformed hamster cells (BHKpy, BHKpygiv, NILpy) and human malignant cells (BT-20 human breast and pancreatic carcinoma cells) were killed by the addition of as little as 0.5 mug of acceptor (per ml of medium), while nontransformed counterparts did not show a significant change in growth or morphology. In vivo studies showed that the acceptor inhibited development and progression of tumors in hamsters inoculated with tumorigenic BHKpy cells. Growth of tumors was inhibited 69-94% in animals given 20 mug of acceptor subcutaneously and 39-67% when acceptor was given intraperitoneally at the time of tumor cell inoculation. Administration of the acceptor after the development of a palpable tumor ( approximately 0.5 cm) caused a 60-85% reduction in growth rate and, in some cases, actual reduction in size and disappearance of palpable tumor. These studies demonstrate that a galactosyltransferase glycopeptide acceptor purified from human malignant effusions produces selective inhibition of transformed cell growth in animal and tissue culture systems.


Tissue-specific changes in molecular clocks during the transition from pregnancy to lactation in mice.
Circadian clocks regulate homeostasis and mediate responses to stressors. Lactation is one of the most energetically demanding periods of an adult female's life. Peripartum changes occur in almost every organ so the dam can support neonatal growth through milk production while homeostasis is maintained. How circadian clocks are involved in adaptation to lactation is currently unknown. The abundance and temporal pattern of core clock genes' expression were measured in suprachiasmatic nucleus, liver, and mammary from late pregnant and early lactation mice. Tissue-specific changes in molecular clocks occurred between physiological states. Amplitude and robustness of rhythms increased in suprachiasmatic nucleus and liver. Mammary rhythms of core molecular clock genes were suppressed. Attenuated rhythms appeared to be a physiological adaptation of mammary to lactation, because manipulation of timing of suckling resulting in significant differences in plasma prolactin and corticosterone had no effect on amplitude. Analysis of core clock proteins revealed that the stoichiometric relationship between positive (CLOCK) and negative (PER2) components remained 1:1 in liver but was increased to 4:1 in mammary during physiological transition. Induction of differentiation of mammary epithelial cell line HC11 with dexamethasone, insulin, and prolactin resulted in similar stoichiometric changes among positive and negative clock regulators, and prolactin induced phase shifts in HC11 Arntl expression rhythm. Data support that distinct mechanisms drive periparturient changes in mammary clock. Stoichiometric change in clock regulators occurs with gland differentiation. Suppression of mammary clock gene expression rhythms represents a physiological adaptation to suckling cues. Adaptations in mammary clock are likely needed in part to support suckling demands of neonates.


The Role of the NLRP3 Inflammasome in Fibrosis.
Fibrosis leads to the deposition of collagens in organs and tissues. The resulting pathology induces a loss of function in the organ it is manifested in and this loss of function modulates the morbidity and mortality in that individual. Indeed, approximately 45% of all deaths in the Western world can be attributed to fibrosis and there are no FDA approved drugs for the treatment of fibrosis. The recent discovery of the inflammasome has led to a plethora of studies investigating this inflammatory signaling pathway in a wide variety of pathogen associated diseases. Many studies have focused on the NLRP3 inflammasome and this inflammasome is activated by a wide variety of cellular alarm signals. Once activated, caspase-1 is cleaved, inducing the secretion of IL-1β and IL-18 that signal to aid in the clearance of invading organisms. However, as the knowledge of the inflammasome has expanded, it was found that it can directly control collagen synthesis, leading to the increased deposition of collagens in the tissues such as the lung, liver, heart, and skin. Mice lacking the inflammasome adaptor protein, ASC, failed to become fibrotic when exposed to bleomycin. Inhibition of caspase-1 activity in fibroblasts from patients with the fibrotic disease systemic sclerosis, decreased collagen synthesis and reduced α-smooth muscle actin expression in myofibroblasts. Taken together, these observations suggest that the inflammasome can drive the fibrotic response and paves the way for novel therapeutics to be identified.


Effects of Body Mass Index on Bone Loading Due to Physical Activity.
The aim of the current study was to compare bone loading due to physical activity between lean, and overweight and obese individuals. Fifteen participants (lower BMI group: BMI < 25 kg/m2, n = 7; higher BMI group: 25 kg/m2 < BMI < 36.35 kg/m2, n = 8) wore a tri-axial accelerometer on 1 day to collect data for the calculation of bone loading. The International Physical Activity Questionnaire (short form) was used to measure time spent at different physical activity levels. Daily step counts were measured using a pedometer. Differences between groups were compared using independent t-tests. Accelerometer data revealed greater loading dose at the hip in lower BMI participants at a frequency band of 0.1-2 Hz (P = .039, Cohen's d = 1.27) and 2-4 Hz (P = .044, d = 1.24). Lower BMI participants also had a significantly greater step count (P = .023, d = 1.55). This corroborated with loading intensity (d ≥ 0.93) and questionnaire (d = 0.79) effect sizes to indicate higher BMI participants tended to spend more time in very light activity, and less time in light and moderate activity. Overall, participants with a lower BMI exhibited greater bone loading due to physical activity; participants with a higher BMI may benefit from more light and moderate level activity to maintain bone health.


Morphological plasticity of the rat supraoptic nucleus--cellular consequences.
The supraoptic nuclei of the hypothalamus display a remarkable anatomical plasticity during lactation, parturition and chronic dehydration, conditions associated with massive neurohypophysial hormone secretion. This structural remodeling is characterized by a pronounced reduction of the astrocytic coverage of oxytocin neurons, resulting in an increase in the number and extent of directly juxtaposed neuronal surfaces. Although the exact role played by such an anatomical remodeling in the physiology of the hypothalamo-neurohypophysial system is still unknown, several findings obtained over the last decade indicate that synaptic and extrasynaptic transmissions are impacted by these structural changes. We review these data and try to extrapolate how such changes at the cellular level might affect the overall activity of the system. One repercussion of the retraction of glial processes is the accumulation of glutamate in the extracellular space. This build-up of glutamate causes an increased activation of pre-synaptic metabotropic glutamate receptors, which are negatively coupled to neurotransmitter release, and a switch in the mode of action of pre-synaptic kainate receptors that control GABA release. Finally, the range of action of substances released from astrocytes and acting on adjacent magnocellular neurons is also affected during the anatomical remodeling. It thus appears that the structural plasticity of the hypothalamic magnocellular nuclei strongly affects neuron-glial interactions and, as a consequence, induces significant changes in synaptic and extrasynaptic transmission.


Long term duration of protective effect for HPV negative women: follow-up of primary HPV screening randomised controlled trial.
To assess whether the increased sensitivity of screening for human papillomavirus (HPV) may represent overdiagnosis and to compare the long term duration of protective effect against cervical intraepithelial neoplasia grade 2 or worse (CIN2+) in HPV based and cytology based screening. 13 year follow-up of the Swedescreen randomised controlled trial of primary HPV screening. Organised cervical screening programme in Sweden. 12,527 women aged 32-38 attending organised screening were enrolled and randomised to HPV and cytology double testing (intervention arm, n=6257) or to cytology only, with samples frozen for future HPV testing (control arm, n=6270). Cumulative incidence of CIN2+ and CIN3+ (Kaplan Meier curves). Longitudinal test characteristics were calculated for cytology only, HPV testing only, and cytology and HPV testing combined, adjusting for censoring. The increased detection of CIN2+ in the intervention arm decreased over time. After six years, the cumulative incidence of CIN3+ was similar in both trial arms, and after 11 years the cumulative incidence of CIN2+ became similar in both arms. The longitudinal sensitivity of cytology for CIN2+ in the control arm at three years was similar to the sensitivity of HPV testing in the intervention arm at five years of follow-up: 85.94% (95% confidence interval 76.85% to 91.84%) v 86.40% (79.21% to 91.37%). The sensitivity of HPV screening for CIN3+after five years was 89.34% (80.10% to 94.58%) and for cytology after three years was 92.02% (80.59% to 96.97%). Over long term follow-up, the cumulative incidence of CIN2+ was the same for HPV screening and for cytology, implying that the increased sensitivity of HPV screening for CIN2+ reflects earlier detection rather than overdiagnosis. The low long term risks of CIN3+ among women who tested negative in HPV screening, support screening intervals of five years for such women. Clinicaltrials.gov NCT00479375.


Characterisation of an epidemic strain of Klebsiella and its variants by computer analysis.
An outbreak of hospital-acquired klebsiella infection was investigated by means of six different typing schemes. Interpretation of the information generated by these schemes proved difficult and allowed only tentative conclusions to be reached concerning the origin of some strains. Consequently, the results were subjected to computer analysis by means of a numerical taxonomic programme adapted for the purpose. A dendrogram was constructed giving the percentage similarity between strains. It established the source of four atypical strains and showed that three of them were derived from the original clone. This form of numerical epidemiological analysis could have useful applications in the investigation of hospital-acquired infection.


Localized whistlers in magnetized spin quantum plasmas.
The nonlinear propagation of electromagnetic (EM) electron-cyclotron waves (whistlers) along an external magnetic field, and their modulation by electrostatic small but finite amplitude ion-acoustic density perturbations are investigated in a uniform quantum plasma with intrinsic spin of electrons. The effects of the quantum force associated with the Bohm potential and the combined effects of the classical as well as the spin-induced ponderomotive forces (CPF and SPF, respectively) are taken into consideration. The latter modify the local plasma density in a self-consistent manner. The coupled modes of wave propagation is shown to be governed by a modified set of nonlinear Schrödinger-Boussinesq-like equations which admit exact solutions in form of stationary localized envelopes. Numerical simulation reveals the existence of large-scale density fluctuations that are self-consistently created by the localized whistlers in a strongly magnetized high density plasma. The conditions for the modulational instability (MI) and the value of its growth rate are obtained. Possible applications of our results, e.g., in strongly magnetized dense plasmas and in the next generation laser-solid density plasma interaction experiments are discussed.


Mapping of Type 1 Diabetes Mellitus.
Type 1 diabetes mellitus (T1DM), an autoimmune disorder, is becoming widespread with approximately 97,700 children in India and 490,000 children worldwide affected. There are various etiological factors contributing to the expansion of its incidence on different geographical locations. Hence, the articles published in reputed journals were studies and data were collected for analyzing the etiology and prevention of T1DM. It has been observed that hybrid insulin peptides act as key antigens for the autoreactive T cells and cause the loss of self-tolerance in humans. The association of coxsackievirus B has been observed with the onset of T1DM. Accurate identification of the trigger can lead to the development of appropriate preventive measures. It can become a base for advance studies to prevent T1DM in humans. This review will highlight the causes and some preventive actions which can be considered to eliminate T1DM.


Association of sexual experience with isolation of various bacteria in cases of ovine epididymitis.
Testicles from rams in flocks experiencing ram epididymitis in Idaho and eastern Oregon were cultured. Twenty-six breeding rams from 6 flocks were cultured and only Brucella ovis was isolated. Virgin rams (65) harbored numerous species of small fastidious gram-negative rods, including Actinobacillus actinomycetemcomitans, Haemophilus spp, Moraxella spp, and Pasteurella spp. Thus, there appeared to be 2 separate disease entities, dependent on sexual experience of the animal.


Competitive uptake and phytomonitoring of chlorinated contaminant mixtures by Redosier dogwood (Cornus sericea).
Plant uptake is an important process in phytoremediation. The robust uptake of volatile organic compounds (VOCs) by plants offers opportunities to establish quantitative relationships between VOCs in plant tissues and in groundwater for the purpose of phytoscreening or phytomonitoring. Most previous research pertaining to phytoremediation neglected the competitive effects of co-contaminants on the uptake of VOCs by plants, yet recent studies appeared to indicate high competitive effects of co-contamination. This study investigated the competitive uptake of three chlorinated compounds in the presence and absence of other co-contaminants by Redosier dogwood in a greenhouse and examined the implications of this competitive phenomenon for phytomonitoring of contaminant mixtures in groundwater. Concentrations of VOCs in stems decreased along the height in both single and bi-solute systems, in agreement with previous observations in the literature. Examination of the VOCs in single and bi-solute systems showed that concentrations of individual compounds are comparable in single and bi-solute systems, yet the ratios of contaminants along the height in bi-solute systems revealed interesting trends. TCE/PCE ratio increased along height while TCE/1,1,2-TCA ratio was roughly constant. The result indicated that sampling point as well as the physicochemical properties of co-contaminants is highly important in phytomonitoring of contaminant mixtures.


Assessment of TGF-beta1-mediated growth inhibition of HPV-16- and HPV-18-transfected foreskin keratinocytes during and following immortalization.
The responsiveness to transforming growth factor-beta1 (TGF-beta1) of two human keratinocyte cell lineages (FK16A and FK18B) generated after transfection with HPV-16 and HPV-18, respectively, was investigated. Both cell lineages revealed loss of heterozygosity (LOH) at 18q and/or 3p associated with the acquisition of the immortal phenotype. These loci harbour genes (TGF-beta receptor II gene at 3p, and Smad2 and Smad4 genes at 18q) encoding products involved in the TGF-beta1 signalling pathway. Mortal and early immortal stages of both cell lineages displayed growth reduction upon exposure to TGF-beta1 concentrations in the range 100 pg/ml to 1 ng/ml. However, the late immortal stages were resistant to TGF-beta1 at concentrations up to 10 ng/ml. TGF-beta1 receptors type I and II were expressed at all stages in both cell lineages. Moreover, mRNA levels of Smad2 and Smad4 genes were nearly constant throughout. TGF-beta1 expression and secretion, which were demonstrated in all analysed stages, may provide selective conditions underlying unresponsiveness to TGF-beta1 upon prolonged monolayer culturing. Thus, LOH at 3p and/or 18q seen during HPV-mediated immortalization of human keratinocytes was not associated with resistance to TGF-beta1-mediated growth inhibition or a marked reduction in TGF- beta1 receptors and mRNA levels of Smad2 or Smad4. Therefore, alternative events are likely to underlie unresponsiveness to TGF- beta1 in late-passage FK16A and FK18B cells.


Evidence in liver for a disulphide-linked scavenger receptor containing a binding site for acetylated low-density lipoprotein and maleylated bovine serum albumin.
Membranes from rat liver were analysed under reducing conditions. The components of the soluble membranes responsible for the binding of acetylated low density lipoprotein (acetyl-LDL) and maleylated bovine serum albumin (Mal-BSA) were chromatographed on a polyethyleneimine-cellulose column and subsequently separated by gel electrophoresis. For both ligands a major binding protein (Mr = 35,000) was revealed by ligand blotting. A minor protein (Mr greater than 67,000) exhibited little binding. The Scatchard plot of the 131I-Mal-BSA binding data of the 35 kDa protein was linear, with a Kd of 17.3 nM. High concentrations of acetyl-LDL competed for half of the 131I-Mal-BSA binding. Excessive Mal-BSA competed for all the visible acetyl-LDL binding. The findings indicate the existence, in the reduced hepatic membrane, of a 35 kDa protein that has two binding sites for 131I-Mal-BSA and one binding site for acetyl-LDL.


WNT5A-ROR2 axis mediates VEGF dependence of BRAF mutant melanoma.
Despite recent advances, approximately 50% of patient with metastatic melanoma eventually succumb to the disease. Patients with melanomas harboring a BRAF mutation (BRAFMut) have a worse prognosis than those with wildtype (BRAFWT) tumors. Unexpectedly, interim AVAST-M Phase III trial data reported benefit from adjuvant anti-VEGF bevacizumab only in the BRAFMut group. We sought to find mechanisms underpinning this sensitivity. We investigated this finding in vitro and in vivo using melanoma cell lines and clones generated by BRAFV600E knock-in on a BRAFWT background. Compared with BRAFWT cells, isogenic BRAFV600E clones secreted more VEGF and exhibited accelerated growth rates as spheroids and xenografts, which were more vascular and proliferative. Recapitulating AVAST-M findings, bevacizumab affected only BRAFV600E xenografts, inducing significant tumor growth delay, reduced vascularity and increased necrosis. We identified 814 differentially expressed genes in isogenic BRAFV600E/BRAFWT clones. Of 61 genes concordantly deregulated in clinical melanomas ROR2 was one of the most upregulated by BRAFV600E. ROR2 was shown to be RAF-MEK regulated in BRAFV600E cells and its depletion suppressed VEGF secretion down to BRAFWT levels. The ROR2 ligand WNT5A was also overexpressed in BRAFMut melanomas, and in ROR2-overexpressing BRAFV600E cells MEK inhibition downregulated WNT5A and VEGF secretion. These data implicate WNT5A-ROR2 in VEGF secretion, vascularity, adverse outcomes and bevacizumab sensitivity of BRAFMut melanomas, suggesting that this axis has potential therapeutic relevance.


[Risk Assessment of Trihalomethane Production Using the Beijiang River and the Pearl River, Guangzhou as Drinking Water Sources].
In order to investigate the risk of trihalomethane formation potential (THMFP) in finished waters as drinking water sources, 70 samples, 114 samples, and 70 samples were collected in November 2013, April 2014 and July 2014, respectively from different locations in the Beijiang River and the Pearl River. After filtration by 0.45 μm filter membrane, a total of 254 samples were chlorinated using Uniform Formation Condition (UFC) method for determining their THM Formation Potential (THMFP). The cancer risk and non-cancer risk of THMs were estimated using USEPA risk assessment model while dominant factors for total risk potential were estimated using sensitivity analysis. Among four THM species, chloroform( CF) was the highest ranging from 101.92-2 590.85 μg x L(-1), followed by bromodichloromethane (BDCM), dibromochloromethane (DBCM) and bromoform (BF). Chloroform, the major THMs speciation, accounted for 96.17% of total THMs. Non-cancer and cancer risk from ingesting THMs was estimated. The result indicated that non-cancer risk of THMs level ranged from 2.03 x 10(-7) to 1.00 x 10(-5) and was not more than 1.0 x 10(-5), the minimum or negligible non-cancer risk level defined by the USEPA. The average cancer risk of THMs was 2.91 x 10(-4) for male and 3.30 x 10(-4) for female in the two rivers, respectively, exceeding the minimum or negligible risk level defined by the USEPA (1. 0 x 10 ~6). The difference of cancer risk between the two rivers was that BDCM ranging from 2.50 x 10(-5) to 6.37 x 10(-4) was approximately twice that of CF in Beijing River. BDCM played an important role in the total risk in the Beijiang River while CF played an important role in the total risk in the Pearl River, Guangzhou. Sensitivity analysis showed that CF played an important role in the estimation of total risk potential, and that the direct utilization of water sources from Beijiang River and the Pearl River Guangzhou is dangerous, thus pretreatment is necessary before chlorination.


Computer-assisted customized antimicrobial dosages.
The use of a computer-based consultation program to customize dosage regimens of antimicrobials for patients with meningitis or bacteremia is described. Using clinical and laboratory information entered by the user, the program determines causative organisms, recommends therapeutic regimens, and generates a graph depicting the expected blood level of each drug as a function of time. During therapy selection, the program considers the site of infection, the susceptibility of the organism to antibiotics, and the patient's clinical status and drug history. Individualized pharmacokinetic values allow for dosage adjustments in renal failure and estimation of blood levels. If renal impairment is present, dosage regimens for drugs excreted by the kidneys are adjusted to assure the desired steady-state blood levels. To help in selection of the optimal regimen, estimated blood levels for each regimen are graphed along with the minimum inhibitory concentration for the organism and the toxic level of the drug. A bulit-in knowledge base in conjunction with patient-specific information enables the computer program to determine appropriate treatment specific to a patient's age, renal function, and prior drug reactions.


The Effect of Acute Limb Ischemia on Mortality in Patients Undergoing Femoral Venoarterial Extracorporeal Membrane Oxygenation.
Extracorporeal membrane oxygenation (ECMO) is a life-saving modality increasingly used in the management cardiopulmonary failure. However, ECMO itself is not without major complications. Mortality remains high, and morbidity such as stroke, renal failure, and acute limb threatening ischemia (ALI) are common among surviving patients. We analyzed the effect of one of these complications, ALI, on the survival of patients receiving venoarterial ECMO (VA ECMO) with femoral cannulation. Patients with cardiopulmonary failure supported by VA ECMO inserted through femoral cannulation at two institutions from December 2010 to December 2017 were enrolled in this study. Data were collected retrospectively. Our primary outcome was ALI and its effect on hospital mortality. Secondary outcomes included six-month mortality, length of hospital stay, and other complications (stroke and renal failure); multivariate logistic regression analysis was used to identify predictors of ALI and hospital mortality. There were 71 patients included in this study. The overall VA ECMO hospital mortality was 53.5%. ALI was seen in 14 (19.7%) patients. Of these, four (5.6%) patients had fasciotomy, four patients (5.6%) had thrombectomy, and one underwent arterial repair (1.4%). Five additional patients (7.0%) with ALI expired and had no vascular intervention. None of the demographic and clinical characteristics significantly correlated with ALI except for stroke and renal failure requiring new-onset hemodialysis (HD). The rate of hospital and 6-month mortality in patients with and without vascular complications were 78.6%, 92.3% and 47.4%, 57.4%, respectively (P = 0.042 and P = 0.023). Multivariate analysis correlated hospital and six-month mortality with ALI, stroke, and new-onset HD. ALI correlates with higher mortality in VA ECMO patients with femoral cannulation. Although some of the contributing factors to mortality in these patients are related to the consequences of cardiopulmonary failure, strong efforts should be made to avoid ALI after femoral VA ECMO cannulation.


Anti-PSMA CAR-engineered NK-92 Cells: An Off-the-shelf Cell Therapy for Prostate Cancer.
Prostate cancer (PCa) has become the most common cancer among males in Europe and the USA. Adoptive immunotherapy appears a promising strategy to control the advanced stages of the disease by specifically targeting the tumor, in particular through chimeric antigen receptor T (CAR-T) cell therapy. Despite the advancements of CAR-T technology in the treatment of hematological malignancies, solid tumors still represent a challenge. To overcome current limits, other cellular effectors than T lymphocytes are under study as possible candidates for CAR-engineered cancer immunotherapy. A novel approach involves the NK-92 cell line, which mediates strong cytotoxic responses against a variety of tumor cells but has no effect on non-malignant healthy counterparts. Here, we report a novel therapeutic approach against PCa based on engineering of NK-92 cells with a CAR recognizing the human prostate-specific membrane antigen (PSMA), which is overexpressed in prostatic neoplastic cells. More importantly, the potential utility of NK-92/CAR cells to treat PCa has not yet been explored. Upon CAR transduction, NK-92/CAR cells acquired high and specific lytic activity against PSMA-expressing prostate cancer cells in vitro, and also underwent degranulation and produced high levels of IFN-γ in response to antigen recognition. Lethal irradiation of the effectors, a safety measure requested for the clinical application of retargeted NK-92 cells, fully abrogated replication but did not impact on phenotype and short-term functionality. PSMA-specific recognition and antitumor activity were retained in vivo, as adoptive transfer of irradiated NK-92/CAR cells in prostate cancer-bearing mice restrained tumor growth and improved survival. Anti-PSMA CAR-modified NK-92 cells represent a universal, off-the-shelf, renewable, and cost-effective product endowed with relevant potentialities as a therapeutic approach for PCa immunotherapy.


Can national cancer registration support clinical databases and clinical cancer research?
Can national cancer registration support clinical databases and clinical cancer research? The short answer is: Yes, it can. A longer version is: Yes, it can and if it cannot substantially help now, we will have to make sure that it can in the future. In many countries and regions for decades there has been a mandatory registration of new and incident cancers. In many of these countries and regions there is also a rapidly growing interest in keeping clinical databases for clinical audit and research. The discussion about how to marry these two systems is growing, not least because both are time and resource demanding and it is important to get priorities right.


Mechanism of protein excretion by gram-negative bacteria: Pseudomonas aeruginosa exotoxin A.
Excretion of proteins by a cell with a double membrane may involve mechanisms different from secretion across a single membrane. We studied this problem with Pseudomonas aeruginosa exotoxin A. This 68,000-dalton protein was released as rapidly as it was completed; even after short pulse-labeling the cells contained neither the toxin nor a larger precursor. Excretion is evidently cotranslational, since in fractionated lysates the toxin was formed (almost entirely in the mature form) by the membrane-polysome complexes but not by the free polysomes. When the membrane was perturbed by 10% ethanol, the cells stopped excreting the toxin and they accumulated an immunoprecipitable, enzymatically active precursor of 71,000 daltons. The precursor was located entirely in the outer membrane on its outer surface. On removal of the ethanol, the cells again excreted mature toxin, but they did not process or release the previously accumulated precursor. Based on these data, a model for the excretion of exotoxin A is presented.


Online Health Information Seeking: A Review and Meta-Analysis.
Online health information, as an emerging field in health communication research, has attracted close attention from researchers. To identify major determinants of why individuals seek health information online, we conducted a meta-analysis that systematically accumulates the existing research findings. To that end, by integrating three theories or models for examining information-seeking behavior, we developed a theoretical framework for the current meta-analysis that emphasizing psychosocial, instrumental, contextual, and demographic factors. By analyzing the effect sizes from 44 articles representing 54 empirical samples, we found that the quality, trustworthiness, and utility of online health information were the dominant predictors of seeking it and that instrumental factors were more important than psychological ones in determining whether individuals did so. Moreover, the development of information and communication technology, the sampling method, and the type of information sought significantly moderated pairwise relationships between determinants and seeking behavior, whereas culture did not. Herein, we discuss the theoretical implications of our findings as well as directions for future research.


Heat treatment of Brussels sprouts retains their ability to induce detoxification enzyme expression in vitro and in vivo.
The bioactive metabolites of glucosinolates, such as isothiocyanates, contained in cruciferous vegetables have been shown to reduce the risk of cancers through the induction of detoxification enzymes. However, cruciferous vegetables are commonly processed before consumption, significantly altering the phytochemical composition of these vegetables. Compared to freeze-dried Brussels sprouts, oven-dried Brussels sprouts contain low concentrations of glucosinolates (22.14 and 0.85 μmol/g, respectively) and isothiocyanates (3.68 and 0.15 μmol/g, respectively). The effect of oven-dried Brussels sprouts on the expression of detoxification enzymes was evaluated in vitro and in vivo. Treatment of immortalized human hepatoma cells with the aqueous extract from oven-dried Brussels sprouts significantly increased quinone activity (0.5 and 1.5 mg/mL) and the activity of the antioxidant response element (EC50=2.39 mg/mL) and xenobiotic response element (EC50 2.92 mg/mL). C3H/HeJ mice fed a diet containing 20% oven-dried Brussels sprout diets for 2 wk demonstrated significantly higher expression than animals fed a nutrient-matched control diet of CYP1A1, CYP1A2, and epoxide hydrolase in the liver and CYP1A1, CYP1A2, CYP1B1, epoxide hydrolase, UGT1A1, thioredoxin reductase, and heme oxygenase in the lungs. The low concentrations of glucosinolates and isothiocyanates in oven-dried Brussels sprouts suggest that other compounds, such as the Maillard reaction products that are produced during heating, are responsible for the induction of detoxification enzymes in vitro and in vivo. The manner in which cruciferous vegetables are processed prior to consumption has significant effects on what compounds people are exposed to. The presence of glucosinolates or isothiocyanates can be a good indicator of the ability of cruciferous vegetables to induce detoxification enzymes. However, the data presented here demonstrate that while heat processing of Brussels sprouts greatly reduced the concentrations of glucosinolates and isothiocyanates, their ability to induce detoxification enzymes in vitro and in vivo was retained.


Molecular diversity and thrombotic risk in protein S deficiency: the PROSIT study.
The Protein S Italian Team (PROSIT) enrolled 79 protein S (PS) deficient families and found 38 PROS1 variations (19 novel) in 53 probands. Of these, 23 variants were selected for expression in'vitro, to evaluate their role as possible causative variants. Transient expression showed high secretion levels (>75%) for three variants, which were considered neutral. Seven missense and five nonsense variants showed low (<or=11%) expression levels and were classified as severe defects. Intermediate expression was observed for eight variants, which were evaluated by factor Va inactivation assay in order to be globally classified as severe or intermediate. Based on the cumulative data, the hazard ratio associated with causative variants was 4.9 (95% CI: 1.4-17.7) for deep vein thrombosis and/or pulmonary embolism, 5.1 (95% CI: 1.1-23.9) for superficial thrombophlebitis, and 4.8 (95% CI: 1.8-13.0) for any venous thrombosis. The hazard ratio for deep vein thrombosis and/or pulmonary embolism in carriers of severe defects only was 7.4 (95% CI: 1.6-24.1). PROSIT showed that dysfunctional variants causing PS deficiency are more common than expected and confirmed that PS deficiency is associated with increased thrombotic risk, although risk assessment is complicated by molecular heterogeneity.


Stereomicroscopic Evaluation of Sealing Ability of Four Different Root Canal Sealers- An invitro Study.
The most commonly used core material for root canal filling is gutta-percha and as the gutta-percha by itself cannot obturate the complete root canal system, owing to its poor sealing properties hence, a sealer is used in combination with root filling material. Sealer is more important than the core obturating material. Sealer plays a secondary role by merely reinforcing (binding or luting) the gutta-percha to the canal walls, however, it is now confirmed that the sealer has a prime role in sealing the canal by blocking the irregularities between the canal space and the core filling material. To investigate the effectiveness of the apical seal obtained by different sealers used in conjugation with cold lateral condensation technique of obturation using gutta-percha under stereomicroscope. One hundred single-rooted extracted human permanent teeth with a single root canal were used in this in-vitro study. The sealers tested were conventional Zinc oxide eugenol sealer, Apexit, AH-Plus and Roekoseal Automix (RSA). The specimens were examined under a stereomicroscope. For the analysis of data Snedocor's F test for the quality of variances among the experimental group and control group (One-Way ANOVA) were employed. The polydimethylsiloxane endodontic root canal sealer RSA provided a significantly better apical seal followed by AH plus and Apexit whereas conventional zinc oxide eugenol showed the lowest sealing ability. It was concluded that there were statistically significant differences amongst the experimental groups. The shrinkage related to setting and potential dissolution might risk the proper seal of the root canal leading to treatment failure.


New evidence for transmitter role of VIP in the airways: impaired relaxation by a catalytic antibody.
The identity of the transmitter(s) of nonadrenergic, noncholinergic airway smooth muscle relaxation has long been investigated. Recently, nitric oxide (NO) has been proposed as the main, if not the only transmitter. We earlier suggested vasoactive intestinal peptide (VIP) as a candidate transmitter and target for pathogenic catalytic autoantibodies (VIPases) found in certain humans. To re-examine the role of VIP, we studied the airway transport and effects of a model monoclonal antibody (Ab) capable of binding and cleaving VIP. In vitro receptor binding assays indicated the catalytic light chain subunit of the VIPase Ab to inhibit the saturable binding of (Tyr(10-125)I) VIP by guinea pig lung membranes, whereas a catalytically deficient mutant of the Ab light chain was without significant inhibitory activity. Systemically administered IgG preparations of the VIPase Ab accumulated in the airway lavage fluid of guinea pigs at levels close to those in blood, suggesting that the Ab reaches the airways freely. Electrical field stimulation (EFS)-induced relaxations of tracheal strips were weaker and shorter in VIPase-treated animals than in control nonimmune IgG-treated animals. The inhibitory effect of the VIPase was dose-dependent. VIPase-mediated inhibition of EFS-induced relaxation was evident both in the absence and presence of blockade of beta-adrenergic and cholinergic receptors. Thus, circulating VIP binding and cleaving antibodies can reach the airways and attenuate the neurogenic relaxation of guinea pig tracheal smooth muscle, probably by neutralizing endogenously released VIP. The findings support a role for VIP as a major mediator of neurogenic relaxation of guinea pig tracheal smooth muscle. Lack of complete abrogation of relaxation is consistent with a co-transmitter role for NO.


Zernike-Galerkin method: efficient computational tool for elastically deformable optics.
We present the Zernike-Galerkin method, a tool for the discretization of partial differential equations (PDEs) on thin membranes in polar coordinates. The use of a truncated Zernike series as ansatz yields a semianalytical compact and parametric solution of the PDE. We demonstrate its use for the solution of the Poisson equation in polar coordinates, which is the equation of governing a thin strained membrane's deformation, or the flow of heat, both of which are important influences for the deformable membrane design. The obtained solution is directly expressed in terms of the components of the wavefront error, which highly facilitates the formulation of design questions. The method is computational highly efficient due to the sparsity and recursivity of the ansatz, is applicable to other PDEs, and can be efficiently combined with geometric optical and optimization methods. Its application to model a pressure-driven adaptive lens membrane is demonstrated.


Antitumor effects of an immunotoxin made with Pseudomonas exotoxin in a nude mouse model of human ovarian cancer.
An immunotoxin composed of Pseudomonas toxin coupled to an antibody to the human transferrin receptor was evaluated for its effect on ovarian cancer. In the tumor model employed, 60 million human ovarian cancer cells were injected into the peritoneal cavity of an immunodeficient nude mouse. By day 5, cancer cells were implanted and growing in small clusters throughout the peritoneal cavity. On days 5-8, 0.3-2 micrograms of immunotoxin was injected into the peritoneal cavity. Control mice died with malignant ascites at 34-58 days after the implantation of tumor cells, whereas immunotoxin-treated mice lived to 100 days or longer. Irrelevant immunotoxins or antibody alone had no antitumor activity. These findings suggest that intraperitoneal injection of immunotoxins may have a role in the treatment of ovarian cancer.


Proteomics and the microbiome: pitfalls and potential.
Introduction: Human symbiotic microbiota are now known to play important roles in human health and disease. Significant progress in our understanding of the human microbiome has been driven by recent technological advances in the fields of genomics, transcriptomics, and proteomics. As a complementary method to metagenomics, proteomics is enabling detailed protein profiling of the microbiome to decipher its structure and function and to analyze its relationship with the human body. Fecal proteomics is being increasingly applied to discover and validate potential health and disease biomarkers, and Therapeutic Goods Administration (TGA)-approved instrumentation and a range of clinical assays are being developed that will collectively play key roles in advancing personalized medicine. Areas covered: This review will introduce the complexity of the microbiome and its role in health and disease (in particular the gastrointestinal tract or gut microbiome), discuss current genomic and proteomic methods for studying this system, including the discovery of potential biomarkers, and outline the development of clinically accepted protocols leading to personalized medicine. Expert commentary: Recognition of the important role the microbiome plays in both health and disease is driving current research in this key area. A proteogenomics approach will be essential to unravel the biologies underlying this complex network.


Multicellular factor analysis of single-cell data for a tissue-centric understanding of disease.
Biomedical single-cell atlases describe disease at the cellular level. However, analysis of this data commonly focuses on cell-type-centric pairwise cross-condition comparisons, disregarding the multicellular nature of disease processes. Here, we propose multicellular factor analysis for the unsupervised analysis of samples from cross-condition single-cell atlases and the identification of multicellular programs associated with disease. Our strategy, which repurposes group factor analysis as implemented in multi-omics factor analysis, incorporates the variation of patient samples across cell-types or other tissue-centric features, such as cell compositions or spatial relationships, and enables the joint analysis of multiple patient cohorts, facilitating the integration of atlases. We applied our framework to a collection of acute and chronic human heart failure atlases and described multicellular processes of cardiac remodeling, independent to cellular compositions and their local organization, that were conserved in independent spatial and bulk transcriptomics datasets. In sum, our framework serves as an exploratory tool for unsupervised analysis of cross-condition single-cell atlases and allows for the integration of the measurements of patient cohorts across distinct data modalities.


Slow inward Ca current in frog heart: theoretical evidence against a voltage-dependent inactivation.
The validity of a Hodgkin-Huxley type voltage-dependent inactivation of slow inward Ca current (Isi) was tested in frog heart using a computer simulation. The time course of Isi was calculated during the development of a frog atrial action potential (AP). With a time constant of inactivation (tauf) of 55 ms at a membrane potential (Em) of -15 mV, the variation of Isi was biphasic: after a transient increase followed by a decrease to zero, Isi partially "reactivated" (at the beginning of the AP repolarization phase) and then fully deactivated. The "reactivation" phase of Isi developed whether tauf was an increasing, decreasing, U-shaped, or bell-shaped function of Em. The addition of an independent and slower process responsible for the recovery from inactivation only partly suppressed the "reactivation" phase. However, until now there was no experimental evidence supporting such a biphasic variation of Isi during AP repolarization. Thus our results indicate that the Hodgkin-Huxley type model of the voltage-dependence of Isi-inactivation process may not correctly represent the actual behavior of frog cardiac muscle.


Intention Prediction and Human Health Condition Detection in Reaching Tasks with Machine Learning Techniques.
Detecting human motion and predicting human intentions by analyzing body signals are challenging but fundamental steps for the implementation of applications presenting human-robot interaction in different contexts, such as robotic rehabilitation in clinical environments, or collaborative robots in industrial fields. Machine learning techniques (MLT) can face the limit of small data amounts, typical of this kind of applications. This paper studies the illustrative case of the reaching movement in 10 healthy subjects and 21 post-stroke patients, comparing the performance of linear discriminant analysis (LDA) and random forest (RF) in: (i) predicting the subject's intention of moving towards a specific direction among a set of possible choices, (ii) detecting if the subject is moving according to a healthy or pathological pattern, and in the case of discriminating the damage location (left or right hemisphere). Data were captured with wearable electromagnetic sensors, and a sub-section of the acquired signals was required for the analyses. The possibility of detecting with which arm (left or right hand) the motion was performed, and the sensitivity of the MLT to variations in the length of the signal sub-section were also evaluated. LDA and RF prediction accuracies were compared: Accuracy improves when only healthy subjects or longer signals portions are considered up to 11% and at least 10%, respectively. RF reveals better estimation performance both as intention predictor (on average 59.91% versus the 62.19% of LDA), and health condition detector (over 90% in all the tests).


Postsynaptic dorsal column pathway of the rat. I. Anatomical studies.
As one of a series of studies of the ascending spinal cord pathways that might be involved in nociception in the rat, we have examined the projection to the dorsal column nuclei that originates in the spinal cord dorsal horn using the retrograde transport of horseradish peroxidase (HRP). This projection in other animals has been called the postsynaptic dorsal column (PSDC) pathway. Small iontophoretic injections of HRP into the cuneate nucleus (CN) labeled more than 350 neurons in alternate sections within the ipsilateral gray matter of segments C6-8. Fewer than 25 neurons were labeled in L4-6 by injections into CN. Injections of HRP confined to the gracile nucleus (GN) labeled more than 200 neurons within a narrow band extending across the ipsilateral dorsal horn subjacent to substantia gelatinosa of L4-6. Fewer than 10 cells were labeled in C6-8 by such injections. Labeling in lumbar neurons following injections into GN was prevented by transection of the dorsal columns at T10, T8, or C2. Thus, neurons labeled by such injections ascend entirely within the dorsal columns. Lesions of the dorsal columns in C2 reduced the number of labeled neurons in the cervical cord following CN injections by approximately 90%. Combined lesions of the dorsal columns and ipsilateral dorsal lateral funiculus (DLF) reduced the number of cells labeled in C6-8 by approximately 98%. Thus, the majority of labeled neurons in the cervical enlargement project to CN via the dorsal columns; a small secondary component of the cervical projection to CN appears to ascend within the DLF. To compare the relative sizes of the projections to the dorsal column nuclei from PSDC neurons and dorsal root ganglion cells (DRG), labeled neurons were counted in the gray matter of the cervical and lumbar enlargements and the corresponding DRG. In the four animals so examined, PSDC neurons constituted over 38% of the neurons that projected to CN and approximately 30% of the cells that projected to GN. These findings indicate that the PSDC projection of the rat is capable of providing a large somatotopically organized input to the dorsal column nuclei.


Near-infrared fundus autofluorescence in early age-related macular degeneration.
To describe the patterns on near-infrared fundus autofluorescence in eyes affected by early age-related macular degeneration. Cross-sectional observational case series. A total of 84 eyes of 84 patients suffering from early age-related macular degeneration (>63 μm but <125 μm drusen and no-to-mild retinal pigment epithelium abnormalities) were enrolled. Patients underwent best-corrected visual acuity, biomicroscopy, infrared reflectance, short-wavelength fundus autofluorescence, and near-infrared fundus autofluorescence. Eyes were classified according to different patterns of near-infrared fundus autofluorescence. Main outcome was definition of relative prevalence and features of each near-infrared fundus autofluorescence pattern; secondary outcomes were correlation between near-infrared fundus autofluorescence and short-wavelength fundus autofluorescence and between near-infrared fundus autofluorescence patterns and best-corrected visual acuity. Four different patterns of near-infrared fundus autofluorescence identified: normal foveal signal (Pattern A, 7%); normal foveal signal with hyperautofluorescent/hypoautofluorescent spots not involving the fovea (Pattern B, 65.5%); hyperautofluorescent/hypoautofluorescent spots involving the fovea (Pattern C, 15.5%); patchy pattern (Pattern D, 12%). best-corrected visual acuity was lower in eyes with foveal signal alteration (Patterns C and D). Near-infrared fundus autofluorescence pattern in early age-related macular degeneration might be suggestive of visual function deterioration when the fovea is involved. Longitudinal studies are warranted to confirm our preliminary results.


Association between IL-17A, -17F and MIF polymorphisms predispose to CpG island hyper-methylation in gastric cancer.
CpG island hyper-methylation (CIHM) is one of the major events in the gastric carcinogenesis. IL-17A, -17F and MIF have a crucial role in the gastric inflammation and carcinogenesis. Recently, we showed that the genetic polymorphisms of MIF-794-CATT repeat are associated with CIHM status in the non-neoplastic gastric mucosa. Consequently, the CIHM status in the gastric cancer tissue, in relation to IL-17A (-197G>A), -17F (7488T>C), and MIF (-173G>C and -794 tetranucleotide repeats) polymorphisms was investigated. Gastric cancer tissues were obtained from 102 patients. CIHM of p14, p16, DAP-kinase and CDH1 genes were determined by methylation-specific polymerase chain reaction (MSP). CIHM high was defined as three or all CpG islands methylated. We employed the PCR-SSCP (multiplex PCR for IL-17A and -17F) method to detect the gene polymorphisms. We did not find significant association between CIHM status and IL-17F (7488T>C) and MIF (-173G>C) polymorphisms. However, concerning the IL-17A (-197G>A) polymorphism, we found that IL-17A G carrier (GG+GA) held a significantly higher risk of CIHM of p16 (OR=11.22, 95% CI=1.38-91.17, p=0.024) and CIHM high (OR=3.51, 95% CI=1.15-10.68, p=0.027). An association was also found between the 7-CATT repeat carrier (5/7 + 6/7 + 7/7) of the MIF polymorphism (-794-CATT) and reduced risk of CIHM of CDH1 (OR=0.36, 95% CI=0.14-0.92, p=0.032). No association was found between CHIM status and homozygote genotypes of each repeat (-794-CATT 5/5, 6/6, and 7/7). The present results provided evidence that the genetic polymorphisms of IL-17A, and MIF-794-CATT repeat are associated with CIHM status in the gastric cancer. Genetic polymorphisms of IL-17A, and MIF-794-CATT repeat may be involved in methylation-related carcinogenesis in the stomach.


Spinal cord ischemia is multifactorial: what is the best protocol?
Despite the improved understanding of spinal cord anatomy and spinal cord ischemia pathophysiology, the rate of debilitating postoperative paraparesis or paraplegia is still not negligible after procedures for thoracic or thoracoabdominal aortic disease. Single studies have demonstrated the role of different treatment modalities to prevent or treat spinal cord ischemia. A multimodal approach, however, is advocated by most authors. Even after the employment of endovascular techniques become routine, the rate of spinal cord ischemia after treatment of thoracoabdominal aortic pathology remained unchanged over time. Spinal cord ischemia is often treatable by different means that concur to improve indirect spinal perfusion through collateral circulation; it should, therefore, be managed promptly and aggressively due to its potential reversibility. Ongoing technical improvements of non-invasive diagnostic tools may allow a better preoperative assessment of the spinal vascular network and a better planning of both open and endovascular thoracic or thoracoabdominal repair.


Effects of two different 5 weeks training programs on the physical fitness of military recruits.
The purpose of this study was to compare the effects of programmed continuous endurance and relative strength training (CERS) with the basic military physical readiness training (BMPR) on the physical readiness. Croatian military recruits (21.3 ± 1.9 years) were divided in the CERS (N = 48) and BMPR (N = 76) groups. Training sessions were conducted three times a week for a total of 5 weeks. The t-test determined positive training effects for both training programs. Significant positive changes in BMPR group were measured in almost all measured variables. In CERS group, besides the 20 m dash run, statistically significant positive changes were not determined in standing horizontal jump, pull-ups and sit and reach test. The positive training effects in both training groups were achieved because of the low initial physical readiness level of the recruits and a short training period in which the statistically significant differences between programs could not be achieved.


Pilot- and process-scale techniques for cell disruption.
Microorganisms are a source of protein with catalytic and/or biological activity, which are of increasing commercial interest for applications in industry or therapy. For the isolation of intracellular products cell disruption is necessary. In principle, chemical, biological, or physical means may be employed to release proteins from cells. These different approaches are reviewed with special emphasis on scale-up and possible industrial operation. Mechanical devices have been improved considerably in recent years and appear most universally suited to cell disintegration. Chemical extraction or enzymatic lysis offers improved selectivity but requires individual procedures for each product. For a final process design, product yield and cost of the unit operation must be considered as well as the possible implications for the subsequent steps in product recovery, especially on solid/liquid separation.


Structural and Immunologic Properties of the Major Soybean Allergen Gly m 4 Causing Anaphylaxis.
Gly m 4 is the major soybean allergen, causing birch pollen cross allergic reactions. In some cases, Gly m 4-mediated anaphylaxis takes place, but the causative factors are still unknown. Here, we studied the structural and immunologic properties of Gly m 4 to shed light on this phenomenon. We showed that Gly m 4 retained its structure and IgE-binding capacity after heating. Gly m 4 was cleaved slowly under nonoptimal gastric conditions mimicking duodenal digestion, and IgE from the sera of allergic patients interacted with the intact allergen rather than with its proteolytic fragments. Similar peptide clusters of Bet v 1 and Gly m 4 were formed during allergen endolysosomal degradation in vitro, but their sequence identity was insignificant. Animal polyclonal anti-Gly m 4 and anti-Bet v 1 IgG weakly cross-reacted with Bet v 1 and Gly m 4, respectively. Thus, we supposed that not only conserved epitopes elicited cross-reactivity with Bet v 1, but also variable epitopes were present in the Gly m 4 structure. Our data suggests that consumption of moderately processed soybean-based drinks may lead to the neutralizing of gastric pH as a result of which intact Gly m 4 can reach the human intestine and cause IgE-mediated system allergic reactions.


Reference Accuracy in Research Quarterly for Exercise and Sport: A 30-Year Follow-Up to Stull et al. (1991).
Purpose: The purpose of this study was to determine the reference accuracy of articles from a random sample of issues from 1999, 2009, and 2019 of Research Quarterly for Exercise and Sport (RQES). Methods: Up to two issues each of RQES published in 1999, 2009, and 2019 were randomly sampled, and references for articles were verified for accuracy. A total of 1,341 references from 50 articles were verified: 1999 (439 references), 2009 (396 references), and 2019 (506 references). Results: The overall error rate was 40% across the three years (i.e., number of references with at least one error divided by the total number of references and multiplied by 100). Chi-square analyses indicated no significant differences between years for the overall error rate (p > .05): 44% (1999), 38% (2009), and 39% (2019). Chi-square analyses indicated significant differences between years for at least one reference with an error related to author(s) names (p > .05). Chi-square analyses indicated significant differences between years for at least one reference with an error related to the article title. Conclusions: In the author's opinion, an overall error rate of 40% across the three years for RQES is simply too high. Ultimately, it is authors' responsibility to ensure reference accuracy. However, given the unacceptable error rates across fields of study, journals, and publication years, it may be prudent to explore supplemental options to increase reference accuracy in peer-reviewed articles. Reference accuracy is an important issue for authors, editorial offices of journals, and the scientific community.


Clinical efficacy and safety of combination therapy of tocilizumab and steroid pulse therapy for critical COVID-19 in HD patients.
Critical coronavirus disease 2019 (COVID-19) has a high fatality rate, especially in hemodialysis (HD) patients, with this poor prognosis being caused by systemic hyperinflammation; cytokine storms. Steroid pulse therapy or tocilizumab (TCZ) have insufficient inhibitory effects against cytokine storms in critical cases. This study evaluated the clinical effects and safety of combining steroid pulse therapy and TCZ. From September 2020 to May 2021, 201 patients with COVID-19 were admitted to our hospital. Before February 2021, patients with an oxygen demand exceeding 8 L/min were intubated and treated with standard therapy (dexamethasone and antiviral therapy). After February 2021, patients underwent high-flow nasal cannula oxygen therapy and were treated with TCZ (8 mg/kg) and methylprednisolone (mPSL) (500 mg/day [≤ 75 kg], 1000 mg/day [> 75 kg]) for 3 days. We compared background characteristics, laboratory findings, and prognosis between non-HD and HD patients and between patients who received and did not receive TCZ and mPSL pulse therapy. Among non-HD patients, the TCZ + mPSL pulse group had significantly higher survival rates and lower secondary infection rates (p < 0.05), than the standard therapy group. All HD patients in the standard therapy group with oxygen demand exceeding 8 L/min died. Contrastingly, all patients in the TCZ + mPSL pulse group survived, with their oxygen demand decreasing to 0-1 L/min within 3 weeks post-administration. TCZ combined with mPSL pulse therapy improved the survival rate without significant adverse events in critical HD and non-HD patients with COVID-19 by strongly suppressing systemic hyperinflammation.


Home intravenous therapy down under.
Health service research traditionally has relied on quantitative methodologies that view the question from the researcher's perspective. Exploring the question of what it is like to receive intravenous therapy at home has required investigation into what is known about home i.v. therapy and the impact on the patient in particular. The purpose of this study was to describe and understand the recipient's lived experience of home i.v. therapy using a research approach that preserves the uniqueness of the experience from the recipient's perspective. The method involved conducting in-depth interviews with a sample of 26 recipients of home i.v. therapy throughout Tasmania, Australia. Interviews were analyzed using an interpretative process, with the participants describing a process that included the main themes of "having a life" and "the hazards of hospitalization." The main goals of the work are to heighten nurses' and other healthcare professionals' understanding and awareness of home i.v. therapy from the consumer perspective and to address the lack of patient/consumer advocacy and exploration within the evaluative literature available on this subject.


Safety, efficacy, and effects on glycemic control of electroconvulsive therapy in insulin-requiring type 2 diabetic patients.
To determine the safety, efficacy, and effects on glycemic control of electroconvulsive therapy (ECT) in insulin-requiring type 2 diabetic patients. Chart review was conducted of 19 patients with insulin-requiring type 2 diabetes mellitus who underwent ECT at Mayo Clinic (Rochester, Minnesota) between 1993 and 2000. None of the patients received oral hypoglycemic drugs. Daily fasting blood glucose concentrations and daily insulin requirements were determined. For each patient, the daily insulin requirements before the first ECT treatment were compared with insulin requirements after the last ECT treatment. In addition, the insulin requirements 1 day before and 1 day after each ECT treatment (n = 143) were compared. Charts were reviewed for activity, appetite, and weight changes. The average change in daily insulin requirements 1 day before the initiation of ECT compared with 1 day after the completion of ECT was not statistically significant. Changes in dietary or activity levels correlated with fluctuations in insulin requirements in selected individuals. There were no significant changes in acute glycemic control associated with ECT. Our findings refute previous reports of dangerous hyperglycemia or improved glycemic control due to ECT in insulin-requiring type 2 diabetes, and suggest that ECT is safe and efficacious in these patients.


[Antihypoxic and antiamnesic effects of mexidol and semax].
Upon single administration, mexidol and semax only in doses of 100 and 0.05 mg/kg, respectively, produced an antihypoxic effect on mice in the altitude chamber and hermetic chamber tests. Preventive course administration of mexidol and semax for 6 days gave significant antihypoxic effect on the model of acute hypobaric hypoxia in mice in doses of 75 and 0.1 mg/kg per day, respectively, in which the same preparations upon single administration were ineffective. Neither mexidol nor semax upon single administration were effective on the models of acute hemic and histotoxic hypoxia. On various models of amnesia (except that induced by the maximal electroshock) in mice, both mexidol and semax exhibited marked antiamnesic effects comparable with that of the reference nootrope drugs piracetam and oxyracetam. Mexidol showed a linear, while semax exhibited a bell-shaped reversible dose-effect relationships. Mexidol and semax inhibited the ortho- and antidromic population response spikes of CA1 pyramidal neurons of survival hippocampal slices in rats. It was estimated that mexidol (in contrast to semax) increased oxygen consumption in rat brain mitochondria and had a linear dose-effect relationship in a concentration range of 1-5 mM. It is concluded that mexidol should be used in high doses (for both single and course administration) for obtaining antihypoxic and antiamnesic effects, while semax requires a thoroughly controlled choice of dosage.


Natural and antibody-dependent cell-mediate cytotoxicity to cultured target cells superinfected with Epstein-Barr virus.
The relationship between natural cell-mediated cytotoxicity (NCMC) and antibody-dependent cell-mediated cytotoxicity (ADCC) was examined in an Epstein-Barr virus-infected target cell system. The total ADCC reactivity to Epstein-Barr virus-infected target cells varied considerably with different effector cells, indicating contributions to specificity by the effector cells as well as by antibodies in the sera. To investigate the role of each reactant, the effector cells, sera, and target cells were tested according to a three-dimensional experimental design and examined for selectivity with the two- and three-way interaction analysis. The two-way analysis was applied to different planes from the experiment to examine special interactions involving two of the three reactants. Selective ADCC was examined through the results from sera versus target cells, selective NCMC was examined by effector cells versus target cells, and the relationship between NCMC and ADCC was examined through the final plane of effector cells versus sera. A three-way interaction analysis applied to the same results supported the conclusions from the two-way analysis and allowed further inquiry into the concurrent role of three reactants. The design and analysis used in the study allowed detection of selective ADCC and NCMC for Epstein-Barr virus-infected target cells and variations in the efficiency of ADCC by different effector cells and in the modulation of NCMC by different sera.


Polypharmacy Among Adults Aged 65 Years and Older in the United States: 1988-2010.
Older adults frequently have several chronic health conditions which require multiple medications. We illustrated trends in prescription medication use over 20 years in the United States, and described characteristics of older adults using multiple medications in 2009-2010. Participants included 13,869 adults aged 65 years and older in the National Health & Nutrition Examination Survey (1988-2010). Prescription medication use was verified by medication containers. Potentially inappropriate medications were defined by the 2003 Beers Criteria. Between 1988 and 2010 the median number of prescription medications used among adults aged 65 and older doubled from 2 to 4, and the proportion taking ≥5 medications tripled from 12.8% (95% confidence interval: 11.1, 14.8) to 39.0% (35.8, 42.3).These increases were driven, in part, by rising use of cardioprotective and antidepressant medications. Use of potentially inappropriate medications decreased from 28.2% (25.5, 31.0) to 15.1% (13.2, 17.3) between 1988 and 2010. Higher medication use was associated with higher prevalence of functional limitation, activities of daily living limitation, and confusion/memory problems in 2009-2010, although these associations did not remain after adjustment for covariates. In multivariable models, older age, number of chronic conditions, and annual health care visits were associated with increased odds of using both 1-4 and ≥5 medications. Additionally, body mass index, higher income-poverty ratio, former smoking, and non-black non-white race were associated with use of ≥5 medications. Prescription medication use increased dramatically among older adults between 1988 and 2010. Contemporary older adults on multiple medications have worse health status compared with those on less medications, and appear to be a vulnerable population.


Relationship between joint motion and flexor tendon force in the canine forelimb.
To increase in vivo tendon force and gliding after flexor tendon repair, a variety of modifications to the methods by which protective passive motion is administered have been advocated. To determine the relationship between the prime variables, wrist and digital position, muscle activation, and in vivo tendon force, a clinically relevant canine model was developed. Force was measured in the flexor tendon during several joint manipulation paradigms: single-finger flexion-extension with the wrist flexed (group 1F), single-finger flexion-extension with the wrist extended (group 1E), four-finger flexion-extension with the wrist flexed (group 4F), four-finger flexion-extension with the wrist extended (group 4E), and synergistic wrist and finger motion where wrist extension and finger flexion were performed simultaneously, followed by wrist flexion and finger extension (group SYN). In addition, tendon force was measured during electric stimulation of the proximal flexor muscle mass. Passive tendon force with the wrist extended (groups 1E and 4E) was two to three times greater than that measured with the wrist flexed, independent of the number of digits moved. With the wrist extended, peak tendon force reached 1,997 g +/- 194 g during single-digit manipulation (group 1E), compared to only 853 g +/- 104 g with the wrist flexed during the same maneuver (group 1F). Statistical comparison between means revealed that groups 1E and 4E were significantly different from groups 1F, 4F, and SYN (p < .005). There were no significant differences between groups 1E and 4E or between groups 1F, 4F, and SYN (p > .200). Active muscle force elicited by electrical stimulation and passive force varied dramatically as the wrist was flexed from full extension 3460 g +/- 766 g to full flexion 427 g +/- 239 g (p < .001). Simultaneously, passive tension decreased from 940 g +/- 143 g with wrist extended to 76 g +/- 37 g with the wrist flexed. These data indicate that wrist position has the greatest effect on flexor tendon force during motions that are commonly used to rehabilitate flexor tendon repairs. Thus, if force is to be controlled during passive motion, wrist-joint angle will have the dominant effect, while the number of digits manipulated will have much less of an effect. If the clinical goal is to minimize tendon force, rehabilitation could be carried out with the wrist flexed, whereas if the goal is to increase tendon force, rehabilitation could include exercise programs that use a greater degree of wrist extension.


The AMPK Activator A769662 Blocks Voltage-Gated Sodium Channels: Discovery of a Novel Pharmacophore with Potential Utility for Analgesic Development.
Voltage-gated sodium channels (VGSC) regulate neuronal excitability by governing action potential (AP) generation and propagation. Recent studies have revealed that AMP-activated protein kinase (AMPK) activators decrease sensory neuron excitability, potentially by preventing sodium (Na+) channel phosphorylation by kinases such as ERK or via modulation of translation regulation pathways. The direct positive allosteric modulator A769662 displays substantially greater efficacy than other AMPK activators in decreasing sensory neuron excitability suggesting additional mechanisms of action. Here, we show that A769662 acutely inhibits AP firing stimulated by ramp current injection in rat trigeminal ganglion (TG) neurons. PT1, a structurally dissimilar AMPK activator that reduces nerve growth factor (NGF) -induced hyperexcitability, has no influence on AP firing in TG neurons upon acute application. In voltage-clamp recordings, application of A769662 reduces VGSC current amplitudes. These findings, based on acute A769662 application, suggest a direct channel blocking effect. Indeed, A769662 dose-dependently blocks VGSC in rat TG neurons and in Nav1.7-transfected cells with an IC50 of ~ 10 μM. A769662 neither displayed use-dependent inhibition nor interacted with the local anesthetic (LA) binding site. Popliteal fossa administration of A769662 decreased noxious thermal responses with a peak effect at 5 mins demonstrating an analgesic effect. These data indicate that in addition to AMPK activation, A769662 acts as a direct blocker/modulator of VGSCs, a potential mechanism enhancing the analgesic property of this compound.