Bronchial hyperresponsiveness and adult onset wheeze: the influence of atopy.
The commonly held belief that adult onset wheezing illness is primarily nonatopic in nature suggests that the role of atopy in the pathophysiology of bronchial hyperresponsiveness (BHR) in adult onset wheeze may be minimal. This study examined risk factors for BHR (BHR: provocative dose causing a 20% fall in forced expiratory volume in one second PD20 < or =16.38 micromol methacholine) among 82 subjects with adult onset wheeze and among 191 subjects who had never wheezed. Subjects were identified from a cohort of subjects aged 39-45 yrs who were known to have had no childhood wheeze and who were involved in a 30 yr follow-up survey. Risk factors for BHR were examined among all subjects with BHR and among subjects with BHR stratified according to whether or not they had ever wheezed. The prevalence of BHR was 40% (33/82) among the subjects with adult onset wheeze and 11% (21/191) among the subjects who had never wheezed. Lower baseline lung function (odds ratio (OR) = 0.94; 95% confidence interval (CI) = 0.92-0.97 per unit forced expiratory volume (FEV1)% predicted) and atopy (OR = 7.23; CI = 2.53-20.64 for all three measures of atopic compared to nonatopic) were associated with BHR, while smoking and family history showed no statistically significant relation to BHR. This pattern was also apparent in analyses stratified by symptom status. A family history of atopy increased the risk that BHR was accompanied by wheezing symptoms (OR = 4.75; CI = 1.53-14.72 for more than one affected relative compared to no affected relatives). These findings suggest that atopy is associated with bronchial hyperresponsiveness in adults known to have had no childhood wheeze. A familial factor reflecting genetic influences and/or shared environmental factors may influence whether bronchial hyperresponsiveness is associated with symptoms.


Multidisciplinary perspective of hepatocellular carcinoma: A Pacific Northwest experience.
Hepatocellular carcinoma (HCC) is the most rapidly increasing type of cancer in the United States. HCC is a highly malignant cancer, accounting for at least 14000 deaths in the United States annually, and it ranks third as a cause of cancer mortality in men. One major difficulty is that most patients with HCC are diagnosed when the disease is already at an advanced stage, and the cancer cannot be surgically removed. Furthermore, because almost all patients have cirrhosis, neither chemotherapy nor major resections are well tolerated. Clearly there is need of a multidisciplinary approach for the management of HCC. For example, there is a need for better understanding of the fundamental etiologic mechanisms that are involved in hepatocarcinogenesis, which could lead to the development of successful preventive and therapeutic modalities. It is also essential to define the cellular and molecular bases for malignant transformation of hepatocytes. Such knowledge would: (1) greatly facilitate the identification of patients at risk; (2) prompt efforts to decrease risk factors; and (3) improve surveillance and early diagnosis through diagnostic imaging modalities. Possible benefits extend also to the clinical management of this disease. Because there are many factors involved in pathogenesis of HCC, this paper reviews a multidisciplinary perspective of recent advances in basic and clinical understanding of HCC that include: molecular hepatocarcinogenesis, non-invasive diagnostics modalities, diagnostic pathology, surgical modality, transplantation, local therapy and oncological/target therapeutics.


[Structural organization and Lugaro neuron connections in the cat cerebellar cortex].
Two types of intermedial Lugaro cells are distinguished in the cat cerebellum: fusiform (horizontal, vertical and obliquely oriented at angles 60-70 to longitudinal axis of folia cerebelli at different levels of granular layer) and those with triangular somata, sparsely branching dendrites and small number of spines. Both fusiform and triangular neuron axons do not stretch out beyond the borders of cerebellar cortex and may be thought of as recurrent interneurons as they contain GABA and glycine. Lugaro cells form numerous contacts almost with all cortical nervous elements: piriform (Purkinje) and Golgi cells, granular and basket neurons, parallel and mossy fibres and even unipolar, (brush) cells. At light microscopic level they are represented as axosomatic and axodendritic contacts. Lugaro cells are the only cerebellar cortical interneurons with dendritic arborizarion oriented in horizontal plane in contrast to vertical orientation of the other interneurons of cerebellar cortex.


Immature stages of development in the parasitoid wasp, Diachasmimorpha longicaudata.
The morphological changes experienced during the immature stages of the solitary parasitoid Diachasmimorpha longicaudata (Ashmead) (Hymenoptera: Braconidae: Opiinae) were studied. This natural enemy of several species of tephritid fruit flies is widely used in biological control strategies. Immature stages are poorly understood in endoparasitoids because they exist within the host. In the present work, developmental processes are described for this species, reared in Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) larvae under controlled environmental conditions. At 25 degrees C, 85% RH, and with an 18:6 L:D photoperiod, preimaginal development takes about 16 days. Five preimaginal stages can be described: egg, three larval instars, prepupa, pupa, and pharate adult. Superparasitism was found in 20% of the host pupae, and the number of oviposition scars was positively correlated with the number of parasitoid larvae per host puparium. The results are compared and discussed with previous studies on related species.


Mechanism of action of rifampin on Mycobacterium smegmatis.
Deoxyribonucleic acid (DNA)-dependent ribonucleic acid (RNA) polymerase (EC 220.127.116.11) isolated from a rifampin-sensitive strain of Mycobacterium smegmatis was 90% inhibited by 1 mug of rifampin per ml; enzyme from a rifampin-resistant mutant was not affected by this concentration of antibiotic. Inhibition of phenylalanine-1-(14)C incorporation by rifampin in growing cultures was complete about 6 min after addition of antibiotic. Under the same conditions, uracil-2-(14)c incorporated was blocked after 1.5 to 2 min. Rifampin kills M. smegmatis very slowly. When rifampin-inhibited cultures were transferred to a rifampin-free medium, there was a partial resumption of uracil-2-(14)C incorporation, even in the presence of chloramphenicol. We conclude that a primary event in the inhibition of M. smegmatis by rifampin is the block of DNA-dependent RNA polymerase.


[Physical activity and training against obesity].
A short review deals with the role of physical activity in the prevention and treatment of obesity, in order to avoid cardio-metabolic consequences of accumulation of the fat. Community-level interventions are necessary, based on estimations of the physical activity. Mechanisms and effects of the proper training on the body fat and muscle are summarized, and practical advices are presented for implementation of the regular physical activity.


Beta-nerve growth factor measurements in mouse serum.
Mouse serum beta-nerve growth factor (NGF) levels were measured using a newly developed competitive beta-NGF radioimmunoassay. The basal serum beta-NGF levels in male and female mice were consistently less than 2 ng/ml when these animals were maintained in individual cages for at least 7 days before they were killed. However, in male mice, serum beta-NGF levels were significantly elevated when they were housed 5 per cage. The rise in serum beta-NGF levels, presumably mediated by intermale aggression, was confirmed by grouping previously isolated mice together in one cage for 20 min before they were killed. In all aggressive male mice, serum beta-NGF levels were elevated by two orders of magnitude. The beta-NGF radioimmunoassay values were also validated by a neurite outgrowth bioassay system using the serum of aggressive male mice. In summary, both measurement techniques confirm that mouse serum beta-NGF levels undergo marked changes depending on animal handling conditions.


Screening of Indian aphrodisiac ayurvedic/herbal healthcare products for adulteration with sildenafil, tadalafil and/or vardenafil using LC/PDA and extracted ion LC-MS/TOF.
Ayurvedic/herbal healthcare products are considered safe under the impression that they are derived from natural products. But recently, there have been several reports worldwide on the adulteration of synthetic PDE-5 inhibitors in aphrodisiac herbal formulations. Therefore, the objective of the present study was to explore the presence of synthetic PDE-5 inhibitors (sildenafil, tadalafil and/or vardenafil) in ayurvedic/herbal healthcare products sold in Indian market for aphrodisiac/related uses. In total, 85 herbal formulations (HFs) were included in the study. The formulations were extracted with methanol and subjected to centrifugation. The supernatant was analysed by HPLC and LC-MS/TOF. Early detection of the presence of sildenafil, tadalafil and vardenafil in the herbal samples was done by the study of extracted ion mass chromatograms at the m/z values of respective parent ions, and two prominent fragments of each. In case of sildenafil and tadalafil, adulteration was also detected by comparing the relative retention times (RR(T)) and UV spectra. Further substantiation was done through comparison of accurate mass spectra with those of the two available standards. Of the 85 HFs tested, only one was eventually found to be adulterated with sildenafil. The extent of adulterant in this sample was determined to the therapeutic dose in the formulation. The study thus indicates emergence of the problem of adulteration of Indian herbal products with PDE-5 inhibitors.


Biologics in bowel cancer.
Colorectal cancer is the third most common cancer in the United States and second leading cause of cancer death with over 50,000 patients expected to die from their disease in 2017. For patients who present at diagnosis with advanced disease the standard treatment is systemic chemotherapy. Over the last decade a number of biologic therapies have emerged as viable treatment options for advance colorectal cancer. When these new drugs are combined with chemotherapy survival is prolonged, often without a detriment to quality of life. In this chapter we will review the most active biologic options for treatment of colorectal cancer and place them in the context of a rapidly growing field.


Predictive roles of D-dimer for mortality of patients with community-acquired pneumonia: a systematic review and meta-analysis.
To explore the predictive roles of D-dimer for the mortality of patients with community-acquired pneumonia (CAP). This was a systematic review and meta-analysis. We searched the following databases: PubMed, EMBASE, Web of Science, Ovid MEDLINE, and Cochrane Library from their inception to July 26, 2020. Studies exploring the relationship between blood D-dimer levels and CAP-related mortality were selected. In this meta-analysis, we calculated mortality rates, sensitivity, specificity, positive likelihood ratios, and negative likelihood ratios. The search identified 1,073 articles, 8 of which (a total of 2,126 patients) were included in this meta-analysis. The pooled mortality rate of the overall sample was 0.10 (95% CI, 0.08-0.14). The levels of blood D-dimer in the nonsurvivors were significantly higher than those in the survivors (weighted mean difference = 1.03 mg/L [95% CI, 0.81-1.26]; p < 0.00001). The area under the summary ROC curve for the optimal cutoff value of D-dimer as a predictor of mortality was 0.848 (SE = 0.046), and the pooled negative likelihood ratio for D-dimer within the normal range was 0.24 (95% CI, 0.11-0.53). Blood D-dimer might be helpful for the initial assessment of mortality risk of patients with CAP. D-dimer levels within the normal range indicate low risk of mortality. Because of the small sample size in our study, our findings should be further explored and validated in future studies with larger sample sizes.


Sex differences in n-3 and n-6 fatty acid metabolism in EFA-depleted rats.
We studied the effect of sex on the distribution of long-chain n-3 and n-6 fatty acids in essential fatty acid-deficient rats fed gamma-linolenate (GLA) concentrate and/or eicosapentaenoate and docosahexaenoate-rich fish oil (FO). Male and female weanling rats were rendered essential fatty acid deficient by maintaining them on a fat-free semisynthetic diet for 8 weeks. Thereafter, animals of each sex were separated into three groups (n = 6) and given, for 2 consecutive days by gastric intubation, 4 g/kg body wt per day of GLA concentrate (containing 84% 18:2n-6), n-3 fatty acid-rich FO (containing 18% 20:5n-3 and 52% 22:6n-3), or an equal mixture of the two oil preparations (GLA + FO). The fatty acid distributions in plasma and liver lipids were then examined. GLA treatment increased the levels of C-20 and C-22 n-6 fatty acids in all lipid fractions indicating that GLA was rapidly metabolized. However, the increases in 20:3n-6 were less in females than those in males, while those in 20:4n-6 were greater, suggesting that the conversion of 20:3n-6 to 20:4n-6 was more active in female than in male rats. FO treatment increased the levels of 20:5n-3 and 22:6n-3 and reduced those of 20:4n-6. The increase in n-3 fatty acids was greater in females than that in males and the reduction in 20:4n-6 was smaller. Consequently, the sum of total long-chain EFAs incorporated was greater in females than that in males. The administration of n-3 fatty acids also reduced the ratio of 20:4n-6 to 20:3n-6 in GLA + FO-treated rats indicating that n-3 fatty acids inhibited the activity of delta-5-desaturase. However, this effect was not affected by the sex difference.


Maximum rate of pressure development and maximal relaxation rate of respiratory muscles in patients with cystic fibrosis.
Respiratory muscle function in patients with cystic fibrosis (CF) has been studied by measurement of maximal inspiratory pressure (P(Imax)), maximal expiratory pressure (P(Emax)), and the pressure-time index of the respiratory muscles (PTI(mus)). The maximum rate of pressure development (MRPD) during P(Imax) (MRPD-P(Imax)), MRPD during P(Emax) (MRPD-P(Emax)), maximal relaxation rate (MRR) during P(Imax) (MRR-P(imax)), and MRR during P(Emax) (MRR-P(Emax)) have not been studied in CF. Our aim was to study MRPD and MRR and investigate their possible application as accessory indices of respiratory muscle function in patients with CF. FEVY, FVC, and maximal expiratory flow between 25% and 75% of vital capacity, body mass index (BMI), upper arm muscle area, P(imax), P(Emax), PTI(mus), MRPD-P(Imax), MRPD-P(Emax), MRR-P(imax), and MRR-P(Emax) were assessed in 123 CF patients and in a control group of 123 healthy subjects matched for age and sex. MRR-P(Emax) was significantly increased and MRPD-P(Emax) was significantly decreased in the CF patients, compared to the healthy controls. In the CF patients MRR-P(Imax) was significantly related to PTI(mus) (P = .02), FEV1 (P = .03), FVC (P = .001), BMI (P < .001), and upper arm muscle area (P < .001). In the CF patients, MRPD-P(Imax) and MRPD- P(Emax) were significantly related to upper arm muscle area (P < .001), BMI (P < .001 and P = .01, respectively), P(Imax) (P < .001), and P(Emax) (P < .001). The CF patients exhibited increased MRR and decreased MRPD during maximal respiratory effort, compared to controls. The differences in MRR-P(imax) and MRPD-P(Imax) between the controls and the complete group of CF patients were not significant. MRPD and MRR were significantly related to nutritional and pulmonary function impairment in CF patients. MRPD strongly correlated to maximal respiratory muscle pressures, and MRR strongly correlated to PTI(mus) in patients with CF. These findings suggest that CF patients are at increased risk of respiratory muscle fatigue. Regular determination of MRPD and MRR may be clinically useful in CF patients and help to initiate inspiratory muscle training and noninvasive ventilation.


The presence of musculoskeletal disorders among amateur bowlers.
With increasing numbers of adults taking part in amateur athletic activity during leisure hours there have been indications that amateur athletic activity may have negative effects on musculoskeletal disease. A literature review focusing on musculoskeletal injuries among bowlers provided only limited references. Studying the characteristic of bowlers complaining of musculoskeletal disorders in general and of upper extremity cumulative trauma disorders specifically, and locating the factors that statistically predict musculoskeletal disorders among Israeli bowlers. The research population includes 98 amateur bowlers who attended two bowling clubs (cluster sampling). Musculoskeletal disorders of the research population were assessed by subjective report using the Standardized Nordic Questionnaires. Further use was made of questionnaires concerning bio-demographic data, work assessment, and the characteristics of the bowling game. Sixty two percent of bowlers reported the presence of musculoskeletal symptoms in one or more of their body joints during the preceding year. Regression analysis with the number of painful joints in the upper extremities as a dependent variable revealed one main predictor: the number of leagues in which the bowler participated (beta = 0.22, p < 0.03). When the dependent variable was the number of painful joints in the entire body, the statistical predictor was the average achievement of the bowlers (beta = 0.40, p < 0.001). Research findings shed light on the unique musculoskeletal disorders of bowlers. Bowlers should be instructed with regard to what are the safe limits of their leisure activities.


Functional aspects of redox control during neuroinflammation.
Neuroinflammation is a CNS reaction to injury in which some severe pathologies, regardless of their origin, converge. The phenomenon emphasizes crosstalk between neurons and glia and reveals a complex interaction with oxidizing agents through redox sensors localized in enzymes, receptors, and transcription factors. When oxidizing pressures cause reversible molecular changes, such as minimal or transitory proinflammatory cytokine overproduction, redox couples provide a means of translating the presence of reactive oxygen or nitrogen species into useful signals in the cell. Additionally, thiol-based redox sensors convey information about localized changes in redox potential induced by physiologic or pathologic situations. They are susceptible to oxidative changes and become key events during neuroinflammation, altering the course of a signaling response or the behavior of specific transcription factors. When oxidative stress augments the pressure on the intracellular environment, the effective reduction potential of redox pairs diminishes, and cell signaling shifts toward proinflammatory and proapoptotic signals, creating a vicious cycle between oxidative stress and neuroinflammation. In addition, electrophilic compounds derived from the oxidative cascade react with key protein thiols and interfere with redox signaling. This article reviews the relevant functional aspects of redox control during the neuroinflammatory process.


Social responsiveness, an autism endophenotype: genomewide significant linkage to two regions on chromosome 8.
Autism spectrum disorder is characterized by deficits in social function and the presence of repetitive and restrictive behaviors. Following a previous test of principle, the authors adopted a quantitative approach to discovering genes contributing to the broader autism phenotype by using social responsiveness as an endophenotype for autism spectrum disorder. Linkage analyses using scores from the Social Responsiveness Scale were performed in 590 families from the Autism Genetic Resource Exchange, a largely multiplex autism spectrum disorder cohort. Regional and genomewide association analyses were performed to search for common variants contributing to social responsiveness. Social Responsiveness Scale scores were unimodally distributed in male offspring from multiplex autism families, in contrast with a bimodal distribution observed in female offspring. In correlated analyses differing by Social Responsiveness Scale respondent, genomewide significant linkage for social responsiveness was identified at chr8p21.3 (multipoint LOD=4.11; teacher/parent scores) and chr8q24.22 (multipoint LOD=4.54; parent-only scores), respectively. Genomewide or linkage-directed association analyses did not detect common variants contributing to social responsiveness. The sex-differential distributions of Social Responsiveness Scale scores in multiplex autism families likely reflect mechanisms contributing to the sex ratio for autism observed in the general population and form a quantitative signature of reduced penetrance of inherited liability to autism spectrum disorder among females. The identification of two strong loci for social responsiveness validates the endophenotype approach for the identification of genetic variants contributing to complex traits such as autism spectrum disorder. While causal mutations have yet to be identified, these findings are consistent with segregation of rare genetic variants influencing social responsiveness and underscore the increasingly recognized role of rare inherited variants in the genetic architecture of autism spectrum disorder.


Photoinduced anchoring and micropatterning of macroinitiators on polyurethane surfaces for graft polymerization of antifouling brush coatings.
Poly[3-azido-2-(2-bromo-2-methylpropanoyloxy)propyl methacrylate] (PAzBrMA) was synthesized as the macroinitiator and anchor for a functional polymer brush coating on polyurethane (PU) films. Ring-opening reaction of the epoxide group of poly(glycidyl methacrylate) with sodium azide produced the hydroxyl and azide functional groups. The hydroxyl groups were substituted with 2-bromoisobutyryl bromide to introduce the alkyl halide initiator. For anchoring, ultraviolet irradiation was applied to convert the azide groups of PAzBrMA physically coated on the PU surface into nitrene intermediates. The nitrene groups reacted with hydrocarbon moieties on the PU surface through hydrogen abstraction to form amine linkages. A photomask could then be employed to create a patterned surface during irradiation. Thus, the anchoring of a PAzBrMA macroinitiator can be achieved under mild conditions, without the use of strong solvents and high temperatures, which will swell or degrade the PU substrates. Finally, 2-hydroxyethyl methacrylate (HEMA) and poly(ethylene glycol) methacrylate (PEGMA) are graft-polymerized on the PAzBrMA-anchored PU film by surface-initiated atom transfer radical polymerization. In comparison with the pristine PU surface, the PU surfaces with grafted HEMA and PEGMA brush coatings were effective in reducing bovine serum albumin adsorption (protein fouling), adhesion of Staphylococcus epidermidis and Pseudomonas sp. (microfouling), and barnacle cyprid settlement (macrofouling). The present surface modification approach provides a simple and versatile means for micropatterning and functionalization of the polymer surfaces.


Physicochemical analysis of the hepatitis B virus core antigen produced by a baculovirus expression vector.
The hepatitis B virus particle consists of an envelope carrying the surface antigen of the virus and an internal capsid consisting of the core antigen (HBcAg). The internal capsid contains the circular, partially dsDNA genome and the viral polymerase. Empty core particles have been produced in Spodoptera frugiperda cells using a recombinant baculovirus vector, YM1KTc, that expresses a 21.4K derivative of the HBcAg gene. The particles have been purified to homogeneity by caesium chloride density gradient centrifugation followed by glycerol gradient centrifugation. Physicochemical analysis of the core particles showed that they exhibited a sedimentation coefficient (s20,(0)w) of 82.5S and a diffusion coefficient (D) of 1.28 x 10(-7) cm2/s. The Mr obtained by substitution of these values in the Svedberg equation was 5.8 x 10(6), using a partial specific volume of 0.73 ml/g for the viral protein as estimated from the amino acid composition. The Mr determined from sedimentation equilibrium analyses was 6.3 x 10(6). Spectrophotometric and metabolic labelling analyses failed to detect nucleic acids in the core preparations. The data are at variance with the prediction that cores exhibit a T = 3 symmetry and contain some 180 subunits. The results suggest that the baculovirus-expressed cores may contain up to 300 subunits of HBcAg protein.


Dissecting megakaryocytopoiesis in vivo with toxigenes.
The genetic programs that regulate the commitment of a totipotent stem cell to the megakaryocytic lineage remain poorly defined and require appropriate in vivo models. Using a cell-specific obliteration technique, a transgenic mouse model was produced where perturbations of megakaryocytopoiesis and platelet production may be induced on demand. This was achieved by targeting the expression of the herpes virus thymidine kinase (HSV-tk) to megakaryocytes using the regulatory regions of the gene coding for the alphaIIb gene, an early marker of megakaryocytopoiesis, which encodes the alpha subunit of the platelet integrin alphaIIb beta3. The HSV-tk gene is not toxic by itself, but sensitizes the target cell to the effect of ganciclovir (GCV), leading to the inhibition of DNA synthesis in dividing cells. The programmed eradication of the megakaryocytic lineage was induced by treating transgenic mice bearing the hybrid construct (alphaIIb-tk) with GCV. After 10 days of treatment, the platelet number was reduced by greater than 96.5% and megakaryocytes were not detectable in the bone marrow (BM). After discontinuing GCV, BM was repopulated with megakaryocytes, and the platelet count was restored within seven days. The recovery was accelerated by the administration of interleukin 11. Prolonged GCV treatment induced erythropenia in the transgenic mice. Assays of myeloid progenitor cells in vitro demonstrated that the transgene was expressed in early erythro-megakaryocytic bipotent progenitor cells. The reversibility and facility of this system provide a powerful model to determine both the critical events in megakaryocytic and erythroid lineage development, and for evaluating the precise role that platelets play in the pathogenesis of a number of vascular occlusive disorders.


Early 18F-FDG-PET/CT as a predictive marker for treatment response and survival in patients with metastatic colorectal cancer treated with irinotecan and cetuximab.
To clarify if early reduction in standard uptake value (SUV) could predict metabolic response, radiologic response and overall survival (OS) in patients with metastatic colorectal cancer receiving third-line treatment. Patients were regardless of KRAS status, included in this phase II trial. They were treated with the monoclonal antibody, cetuximab, and the chemotherapeutic drug, irinotecan, every second week. A F18-fluorodeoxy glucose positron emission tomography/computed tomography (FDG-PET/CT) was scheduled before the first and second treatment, respectively, and then after every fourth treatment. Early metabolic response after one treatment and best overall metabolic response was calculated according to EORTC criteria (responders: ≥15% decrease in ∑SUVmax) and PERCIST (responders: ≥30% decrease in SULpeak). Best overall radiologic response was calculated according to RECIST 1.0. By EORTC criteria, early metabolic response predicted partial metabolic response (PMR) with a high positive predictive value (PPV) of 0.875 and a high negative predictive value (NPV) of 0.714. Partial radiologic response was predicted with a low PPV of 0.368 but a high NPV of 1.0. By PERCIST, PMR was predicted with a high PPV of 0.826 and an intermediate NPV of 0.667 and partial radiologic response was predicted with a low PPV of 0.5 but a high NPV of 1.0. Median OS was nearly the same with the two criteria sets; 14.1 months for early metabolic responders and 9.9 months for non-responders using EORTC criteria and 13.5 and 10.1 months, respectively, using PERCIST. With both EORTC criteria and PERCIST, early reduction in FDG uptake was predictive of a later partial metabolic and partial radiologic response to treatment. It was also predictive of significantly longer survival of early metabolic responders compared to non-responders. However, the sensitivities and specificities were not high enough to support clinical routine use.


Effect of provider prompts on adolescent immunization rates: a randomized trial.
Adolescent immunization rates are suboptimal. Experts recommend provider prompts at health care visits to improve rates. We assessed the impact of either electronic health record (EHR) or nurse- or staff-initiated provider prompts on adolescent immunization rates. We conducted a randomized controlled trial, allocating practices in 1 of 2 practice-based research networks (PBRN) to provider prompts or standard-of-care control. Ten primary care practices participated, 5 intervention and 5 controls, each matched in pairs on urban, suburban, or rural location and practice type (pediatric or family medicine), from a PBRN in Greater Rochester, New York (GR-PBRN); and 12 practices, 6 intervention, 6 controls, similarly matched, from a national pediatric continuity clinic PBRN (CORNET). The study period was 1 year per practice, ranging from June 2011 to January 2013. Study participants were adolescents 11 to 17 years attending these 22 practices; random sample of chart reviews per practice for baseline and postintervention year to assess immunization rates (n = 7,040 total chart reviews for adolescents with >1 visit in a period). The intervention was an EHR prompt (4 GR-PBRN and 5 CORNET practice pairs) (alert) that appeared on providers' computer screens at all office visits, indicating the specific immunizations that adolescents were recommended to receive. Staff prompts (1 GR-PBRN pair and 1 CORNET pair) in the form of a reminder sheet was placed on the provider's desk in the exam room indicating the vaccines due. We compared immunization rates, stratified by PBRN, for routine vaccines (meningococcus, pertussis, human papillomavirus, influenza) at study beginning and end. Intervention and control practices within each PBRN were similar at baseline for demographics and immunization rates. Immunization rates at the study end for adolescents who were behind on immunizations at study initiation were not significantly different for intervention versus control practices for any vaccine or combination of vaccines. Results were similar for each PBRN and also when only EHR-based prompts was assessed. For example, at study end, 3-dose human papillomavirus vaccination rates for GR-PBRN intervention versus control practices were 51% versus 53% (adjusted odds ratio 0.96; 95% confidence interval 0.64-1.34); CORNET intervention versus control rates were 50% versus 42% (adjusted odds ratio 1.06; 95% confidence interval 0.68-1.88). In both a local and national setting, provider prompts failed to improve adolescent immunization rates. More rigorous practice-based changes are needed.


[Forensic medical toxicometry of acute carbon monoxide poisoning during alcoholic intoxication].
Data on general toxicity of carbon monoxide (CO) are presented with reference to the gender, age, and the degree of alcoholic intoxication of the affected subjects. Threshold values of carboxyhemoglobinemia for clinical and morphological manifestations of alcoholic intoxication were determined. Expert evaluation of CO toxicokinetics in subjects with alcoholic intoxication revealed informative clinical and morphological signs of carbon monoxide poisoning, the basic parameters of carboxyhemoglobin kinetics, and the sequence of CO-induced clinical and morphological changes during alcoholic intoxication. The data obtained permit to exactly determine the specific cause of death from poisoning with carbon monoxide and ethanol.


Pitfalls in RNA Modification Quantification Using Nucleoside Mass Spectrometry.
ConspectusIn recent years, there has been a high interest in researching RNA modifications, as they are involved in many cellular processes and in human diseases. A substantial set of enzymes within the cell, called RNA writers, place RNA modifications selectively and site-specifically. Another set of enzymes, called readers, recognize these modifications which guide the fate of the modified RNA. Although RNA is a transient molecule and RNA modification could be removed by RNA degradation, a subclass of enzymes, called RNA erasers, remove RNA modifications selectively and site-specifically to alter the characteristics of the RNA. The detection of RNA modifications can be done by various methods including second and next generation sequencing but also mass spectrometry. An approach capable of both qualitative and quantitative RNA modification analysis is liquid chromatography coupled to mass spectrometry of enzymatic hydrolysates of RNA into nucleosides. However, for successful detection and quantification, various factors must be considered to avoid biased identification and inaccurate quantification. In this Account, we identify three classes of errors that may distort the analysis. These classes comprise (I) errors related to chemical instabilities, (II) errors revolving around enzymatic hydrolysis to nucleosides, and (III) errors arising from issues with chromatographic separation and/or subsequent mass spectrometric analysis.A prominent example for class 1 is Dimroth rearrangement of m1A to m6A, but class 1 also comprises hydrolytic reactions and reactions with buffer components. Here, we also present the conversion of m3C to m3U under mild alkaline conditions and propose a practical solution to overcome these instabilities. Class 2 errors-such as contaminations in hydrolysis reagents or nuclease specificities-have led to erroneous discoveries of nucleosides in the past and possess the potential for misquantification of nucleosides. Impurities in the samples may also lead to class 3 errors: For instance, issues with chromatographic separation may arise from residual organic solvents, and salt adducts may hamper mass spectrometric quantification. This Account aims to highlight various errors connected to mass spectrometry analysis of nucleosides and presents solutions for how to overcome or circumnavigate those issues. Therefore, the authors anticipate that many scientists, but especially those who plan on doing nucleoside mass spectrometry, will benefit from the collection of data presented in this Account as a raised awareness, toward the variety of potential pitfalls, may further enhance the quality of data.


[Angiogenesis and vascular endothelial growth factor, role in the pathology of the gastrointestinal tract].
A review of the literature, the authors present information about angiogenesis, as the process of formation of new blood vessels in the organ or tissue from pre-existing vasculature by migration and proliferation of endothelial cells, as well as on the processes of regulation. Presents key regulatory system angiogenesis. The main attention is paid Vascular endothelial growth factor (VEGF). Specially structured data on the role of Vascular endothelial growth factor in human pathology, including diseases of gastrointestinal tract, inflammatory and neoplastic nature.


Facile Surface Modification of Polyamide Membranes Using UV-Photooxidation Improves Permeability and Reduces Natural Organic Matter Fouling.
A new optimized ultraviolet (UV) technique induced a photooxidation surface modification on thin-film composite (TFC) polyamide (PA) brackish water reverse osmosis (BWRO) membranes that improved membrane performance (i.e., permeability and organic fouling propensity). Commercial PA membranes were irradiated with UV-B light (285 nm), and the changes in the membrane performance were assessed through dead-end and cross-flow tests. UV-B irradiation at 12 J·cm-2 enhanced the pure water permeability by 34% in the dead-end tests without decreasing the mono- or divalent ion rejections, as compared with the pristine PA membrane, and led to less fouling by natural organic matter in the cross-flow tests. Scanning electron microscopy (SEM), attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy, and X-ray photoelectron spectroscopy (XPS) confirmed that UV-B irradiation opened the pore structure and created carboxylic and amine groups on the PA surface, leading to increased membrane surface charge and hydrophilicity. Thus, an optimal UV-B dose appears to modify only a thin layer of the PA membrane surface, which favorably enhances the membrane performance. UV-B did not alter the structure, flux, or salt rejection for cellulose triacetate (CTA)-based membranes. While other membrane surface modifications include oxidants, strong acids, and bases, the UV-B facile treatment is chemical-free, thus reducing chemical wastes, and easy to apply in roll-to-roll fabrication processes of PA membranes. The results also showed that a low UV irradiation dose could be applied to PA or CTA membranes for disinfection or photocatalytic oxidation.


Summarizing Evidence of Associations of COVID-19 With a Future Diagnosis of Inflammatory Rheumatic and Musculoskeletal Diseases: A Rapid Review.
Musculoskeletal symptoms are commonly reported following acute COVID-19. It is unclear whether those with musculoskeletal symptoms subsequently develop inflammatory rheumatic musculoskeletal disease (iRMD). This review seeks to identify evidence for an association between acute COVID-19 and subsequent iRMD diagnosis. A rapid review of the literature using a systematic search of Medline, EMBASE and two COVID-19 databases was undertaken until August 2022. Case studies, case series, cross-sectional, case-control, and cohort studies reporting patients with an incident iRMD following COVID-19 were included. Title and abstract screening were conducted by one reviewer and full text screening by two reviewers. Data extraction and quality appraisal were by one reviewer, with a second verifying. Study-type specific critical appraisal tools were used. Results were narratively synthesized. A total of 80 studies were included (69 case reports, 10 case series and 1 cross-sectional study). Commonly reported iRMDs were "reactive arthropathies" (n = 47), "inflammatory arthropathies unspecified" (n = 18), rheumatoid arthritis (n = 12) and systemic lupus erythematosus (n = 11). The cross-sectional study reported 37% of those with COVID-19 developed "post COVID arthritis." Time from diagnosis of COVID-19 to iRMD presentation ranged from 0 to 120 days. Several mechanisms were proposed to explain the association between COVID-19 and iRMD development: autoimmune processes, aberrant inflammatory responses, colonization of joint spaces, direct damage from the severe acute respiratory syndrome coronavirus 2 virus and genetic predisposition. The level of evidence of the studies included in this review was low and the quality generally poor. Prospective observational studies are required to confirm associations and likely impact of post COVID-19 iRMDs at a population level.


Impetigo. Current etiology and comparison of penicillin, erythromycin, and cephalexin therapies.
We attempted to determine the causative bacterial pathogens of impetigo in children in our area, to compare the effectiveness of three frequently used oral antimicrobial treatment regimens, and to correlate the antimicrobial sensitivity of the bacterial isolates with clinical responses to treatment. Seventy-three children with impetigo were randomly assigned to receive penicillin V potassium or cephalexin monohydrate, both administered in dosages of 40 to 50 mg/kg per day, or erythromycin estolate administered in a dosage of 30 to 40 mg/kg per day. All drugs were given in three divided doses for 10 days. Treatment failure was defined as persistence of lesions 8 to 10 days after initiation of drug therapy as determined by examiners blinded to the treatment therapies. Forty-five (62%) cultures showed Staphylococcus aureus only, 14 (19%) showed S aureus and group A beta-hemolytic streptococci, six (8%) showed group A beta-hemolytic streptococci only, and eight (11%) showed no growth or other organisms. Treatment failure occurred in six (24%) of 25 patients treated with penicillin V, one (4%) of 25 patients treated with erythromycin estolate, and no patients treated with cephalexin. We conclude that S aureus is the most common cause of impetigo in children in our study population, that cephalexin is the most effective treatment, that erythromycin estolate is nearly equally effective and may be preferred on a cost-effectiveness basis, and that penicillin V is inadequate for treatment of this infection.


Cytotoxic activity of novel palladium-based compounds on leukemia cell lines.
Effective treatment methods for human leukemia are under development, but so far none of them have been found to be completely satisfactory. It was recently reported that palladium complexes have significant anticancer activity as well as lower toxicity compared with some clinically used chemotherapeutics. The anticancer activities of two novel palladium(II) complexes, [Pd(sac)(terpy)](sac)·4H2O and [PdCl(terpy)](sac)·2H2O, were tested against three human leukemia cell lines, Jurkat, MOLT-4, and THP-1, in comparison with cisplatin and adriamycin. The cytotoxic effect of the drugs was determined using the MTT assay. Cell death was assessed using fluorescein isothiocyanate-annexin/propidium iodide staining for flow cytometry. Furthermore, p53 phosphorylation, poly(ADP-ribose) polymerase cleavage, and Bax and Bcl-2 mRNA levels were examined to elucidate the mechanism of cell death induction. Both complexes exhibited a significant dose-dependent antigrowth effect in vitro. The complexes predominately induced apoptosis, but necrosis was also observed. In-vitro results have shown that palladium(II) complexes may be regarded as potential anticancer agents for treating human leukemia. Therefore, further analysis to determine the putative mechanism of action and in-vivo studies on animal models are warranted.


Hidradenitis suppurativa associated with spondyloarthritis -- results from a multicenter national prospective study.
To determine the prevalence and characterize the inflammatory musculoskeletal symptoms of hidradenitis suppurativa (HS), a chronic inflammatory disease of skin appendages. Patients with HS referred to 3 dermatology university hospital centers were systematically screened for peripheral arthritis, dactylitis, inflammatory back pain, or enthesitis. After careful clinical examination, patients were further classified according to clinical and imaging criteria for spondyloarthritis (SpA) using the Amor, European Spondyloarthropathy Study Group (ESSG), and ASsessment in ankylosing spondylitis (ASAS). We screened 640 patients with HS; 184 had musculoskeletal symptoms. In all, 43 (mean age 39.4 yrs, ± 8.3; 80% women) had arthritis, inflammatory back pain, or enthesitis and were investigated further. Signs of HS preceded the onset of articular symptoms in 39 patients (90%), at a mean interval of 3.6 years. A total of 18 (41%), 24 (55%), and 15 (34%) patients fulfilled the Amor, ESSG, and ASAS criteria, respectively, while synovitis, acne, pustolosis, hyperostosis, and osteitis (SAPHO) syndrome was established in 4 patients. The crude prevalence of SpA in all 640 patients with HS was 3.7% by the ESSG criteria. SpA may occur in patients with HS, with the prevalence in this group exceeding that in the general population. The very short time between skin and joint symptom onset in some cases suggests common pathogenic mechanisms underlying HS and SpA.


Alcoholic liver disease: roles of alcohol and malnutrition.
A toxic effect of alcohol is the principal cause of the development of liver disease in alcoholism. Fatty infiltration of the liver is a consequence of ethanol metabolism due mainly to an increased synthesis and decreased degradation of fatty acids. Mechanisms that have been suggested for ethanol-induced hepatocellular necrosis include centrolobular hypoxia due to an increased oxygen requirement and intracellular accumulation of protein, fat, and water which results in increased cell size. Hepatocellular necrosis, however, may not be a necessary stage in the development of cirrhosis. Chronic ethanol administration increases hepatic collagen deposition, and acute and chronic ethanol administration inhibit liver cell regeneration. Increased humoral and cellular immunological activity to liver tissue and its components may contribute to the persistence of liver disease in the alcoholic. However, only a small proportion of alcoholics and baboons fed alcohol develop cirrhosis, suggesting that other factors, either genetic, environmental, or nutritional, play a role. Malnutrition is common in alcoholics. Liver disease is more common in some malnourished populations, and has been produced by nutrient deficiencies. Decreased dietary intake, as well as malabsorption and alterations in the metabolism of nutrients, are causes of nutrient deficiencies in alcoholism. Some of the effects of alcohol on the liver may be mediated by its actions on nutrient absorption and metabolism.


Ultra-low friction mechanism of highly sp3-hybridized amorphous carbon controlled by interfacial molecule adsorption.
The friction behaviors of highly sp3-hybridized carbon films, including ultra-nanocrystalline diamond and diamond-like carbon materials, strongly depend on the atmosphere. However, the roles of the corresponding molecules in the interfacial bonding characteristics remain a subject of debate. By means of density functional theory calculations, this study aims to fill a knowledge gap about the correlation between the evolving contact quality induced by the adsorption of molecules, and the friction behavior of highly sp3-bonded carbons. The results prove that gas-solid adsorption is responsible for the diversity in friction coefficients of ultra-nanocrystalline diamond and diamond-like carbons in different atmospheres. This study emphasizes the role of terminal states in friction coefficients, and demonstrates that electron lubrication is another available strategy for hydrogenated diamond-like carbons to achieve ultra-low friction. This conclusion is validated by the ultra-low friction coefficient (∼0.009) of hydrogenated diamond-like carbons in a dry nitrogen atmosphere. These findings provide atomic scale descriptions of the surface passivation mechanisms for ultra-nanocrystalline diamond and diamond-like carbons, which contribute to our understanding of their macro-scale friction behaviors.


WAT-on-a-chip integrating human mature white adipocytes for mechanistic research and pharmaceutical applications.
Obesity and its numerous adverse health consequences have taken on global, pandemic proportions. White adipose tissue (WAT) - a key contributor in many metabolic diseases - contributes about one fourth of a healthy human's body mass. Despite its significance, many WAT-related pathophysiogical mechanisms in humans are still not understood, largely due to the reliance on non-human animal models. In recent years, Organ-on-a-chip (OoC) platforms have developed into promising alternatives for animal models; these systems integrate engineered human tissues into physiological microenvironment supplied by a vasculature-like microfluidic perfusion. Here, we report the development of a novel OoC that integrates functional mature human white adipocytes. The WAT-on-a-chip is a multilayer device that features tissue chambers tailored specifically for the maintenance of 3D tissues based on human primary adipocytes, with supporting nourishment provided through perfused media channels. The platform's capability to maintain long-term viability and functionality of white adipocytes was confirmed by real-time monitoring of fatty acid uptake, by quantification of metabolite release into the effluent media as well as by an intact responsiveness to a therapeutic compound. The novel system provides a promising tool for wide-ranging applications in mechanistic research of WAT-related biology, in studying of pathophysiological mechanisms in obesity and diabetes, and in R&D of pharmaceutical industry.


Larynx Trauma and Hyoid Bone Fracture after Bite Injury in Dog: Case Report.
An 8-year-old male Jack Russell crossbreed dog was admitted to our hospital with dyspnea and shock following a dog-bite injury on the ventral neck. Radiographs revealed subcutaneous emphysema and bilateral thyrohyoid bone fractures. Intraoperatively, rupture of both sternohyoid muscles, both hyoepiglotticus muscles, both thyrohyoid muscles, and a partial cranial rupture of the superficial sphincter colli muscle were detected. Part of the epiglottis was detached from the thyroid cartilage. The patient's severed muscles and torn epiglottis were reattached using a simple interrupted suture pattern. Hyoepiglotticus muscles could not be identified. The bilateral thyrohyoid bone fractures were repaired with intraosseous wire suture. A temporary tracheostomy tube and an esophageal feeding tube were placed postoperatively. The dog was discharged after 8 days, re-examined at 2 and 6 months and laryngeal and pharyngeal function were evaluated as normal. To the authors' knowledge, this is the first report of a dog that presented with laryngeal trauma with hyoid bone fracture and acute dyspnea that underwent surgical treatment resulting in an acceptable outcome.


Integrated Puncture Score: force-displacement weighted rind penetration tests improve stalk lodging resistance estimations in maize.
Stalk lodging (breaking of agricultural plant stalks prior to harvest) is a multi-billion dollar a year problem. Rind penetration resistance tests have been used by plant scientists and breeders to estimate the stalk lodging resistance of maize for nearly a hundred years. However, the rind puncture method has two key limitations: (1) the predictive power of the test decreases significantly when measuring elite or pre-commercial hybrids, and (2) using rind penetration measurements as a breeding metric does not necessarily create stronger stalks. In this study, we present a new rind penetration method called the Integrated Puncture Score, which uses a modified rind penetration testing protocol and a physics-based model to provide a robust measure of stalk lodging resistance. Two datasets, one with a diverse array of maize hybrids and one with only elite hybrids, were evaluated by comparing traditional rind penetration testing and the Integrated Puncture Score method to measurements of stalk bending strength. When evaluating the diverse set of hybrids, both methods were good predictors of stalk bending strength (R2 values of 0.67). However, when evaluating elite hybrids, the Integrated Puncture Score had an R2 value of 0.74 whereas the traditional method had an R2 value of 0.48. Additionally, the Integrated Puncture Score was able to differentiate between the strongest and weakest hybrids in the elite hybrid data set whereas the traditional rind penetration method was not. Additional experiments revealed strong evidence in favor of the data aggregation steps utilized to compute the Integrated Puncture Score. This study presents a new method for evaluating rind penetration resistance that highly correlates with stalk bending strength and can possibly be used as a breeding index for assessing stalk lodging resistance. This research lays the foundation required to develop a field-based high-throughput phenotyping device for stalk lodging resistance.


PSMA Radioligand Therapy in Prostate Cancer: Where Are We and Where Are We Heading?
Diagnosis and treatment of prostate cancer are complex and very challenging, being a major health care burden. The efficacy of radioligand therapy with prostate-specific membrane antigen agents has been proven beneficial in certain clinical indications. In this review, we describe management of prostate cancer patients according to current guidelines, especially focusing on the available clinical evidence for prostate-specific membrane antigen radioligand therapy.


State Control in the Substance-Exposed: Fetus I. The Fetal Neurobehavioral Profile: An Assessment of Fetal State, Arousal, and Regulation Competency.
Behavioral states are stable structures of behaviors that become more definable and coordinated with increasing age. With ultrasound we can see the fetus move, breathe, and react to changes in its environment. Ultrasound used in conjunction with Doppler fetal heart rate recording provides behavioral and neurophysiologic data useful in state determination. The Fetal neurobehavioral Profile (FNP) was developed by our group as an assessment of fetal behaviors reflecting CNS integrity in the drug-exposed fetus. The FNP was designed to parallel methods of examining the newborn infant, especially in state-related behaviors. The FNP measures: fetal responsiveness and arousal after environmental perturbation with vibroacoustic stimulation (VAS);habituation to VAS; state recovery; and self-regulation post-VAS. From the behavioral and physiologic recordings, the constructs of state differentiation, organization, and regulation as well as fetal arousal and regulation competency can be measured. Previous work using the FNP showed that those fetuses with abnormal or suspect fetal state regulation demonstrated impaired performance on the NBAS (Am. J. Obstet. Gynecol. 161: 685,1989). To expand these observations, three populations are currently being studied: prenatal nicotine-exposed, prenatal cocaine-exposed, and controls. Data are from 97 women/fetus dyads and a total of 236 FNP at ages 28-30 weeks gestational age, 31-34 weeks gestational age, and > 36 weeks gestational age. Although there are no group differences in the ability to achieve state by 36 weeks, interesting trends emerge: fetuses prenatally cocaine-exposed spend less time in 1F, more time in 4F, and have fewer transitions. At FNP1, fewer cocaine-exposed fetuses had an initial reaction to VAS, whereas fewer nicotine-exposed fetuses habituated. Although the ability to habituate to VAS did not discriminate the cocaine group from the control or nicotine groups, the number of stimuli required for habituation differed between groups: 7 for the cocaine-exposed, 3 for the nicotine, and 5 for the control groups. Thus latency, a measure of arousal, differs among these groups Preliminary data also susggest a correlation of prenatal data with postnatal outcome.


Post-translational modifications of seminal proteins and their importance in male fertility potential.
Introduction: The seminal proteome has been shown to directly influence the male fertile potential. Post-translational modifications (PTMs) are significant changes that play a role in the biological regulation of proteins. Sperm cells are transcriptionally and translationally inactive and these modifications are essential to control protein function.Areas covered: Here we reviewed seven PTMs which importance for male reproductive function investigated in the past decade, namely S-nitrosylation and tyrosine nitration (both occurring by the action of NO), glycosylation, ubiquitination, acetylation, methylation, and SUMOylation. Since they were previously identified in human semen, we focus on their role in sperm function, as well as in physiological and pathophysiological processes which could contribute to the fertility potential. The following keywords were applied: 'post-translational modification', 'sperm', 'semen', 'seminal plasma', 'male infertility', 'nitrosylation', 'nitration', 'histone methylation', 'SUMOylation', 'ubiquitination', 'ubiquitilation', 'glycosylation', and 'acetylation'.Expert opinion: Most biological processes orchestrated by proteins require PTMs for their activation or inhibition. Most of them are dynamic and occur in mature sperm, modulating protein function, thus exerting a significant role in sperm function and fertility. Finally, the study of PTMs should be also addressed in pathophysiological processes, as different clinical conditions are known to alter the proteome.


Plastic surgeon compliance with national safety initiatives: clinical outcomes and "never events".
Venous thromboembolism and surgical-site infection have been identified as preventable complications that are addressed by the National Quality Forum and the Surgical Care Improvement Project. The authors examined compliance of faculty with venous thromboembolism and surgical-site infection prophylaxis and incidence of adverse outcomes in patients at risk. The authors performed retrospective chart reviews on 243 patients who underwent abdominoplasty or panniculectomy from 2000 to 2007 and documented demographics and adverse outcomes. Analysis was completed using Pearson's chi-square and Fisher's exact test for categorical variables. Significance was set at p < 0.05. Obesity was defined as body mass index more than 30 and morbid obesity was defined as body mass index more than 40. Of 243 patients, 144 (59 percent) were obese. Seventeen patients (7 percent) suffered complications. All 243 patients received at least one form of venous thromboembolism prophylaxis. One patient had a deep venous thrombosis, and two had pulmonary embolism. These three patients were morbidly obese. Seventy-four percent of patients received appropriate antibiotics. Thirteen patients (5.3 percent) developed significant postoperative infection requiring hospitalization, 12 (92 percent) of whom received appropriate antibiotics. Eleven of these 13 patients (85 percent) were obese, and seven (54 percent) were morbidly obese. Obesity proved to be the only significant risk factor (p > 0.05). Despite very good compliance with safe practice initiatives, significant adverse outcomes occurred. Obesity was the only pervasive risk factor. This study highlights the potential need for compliance with quality measures and demonstrates that adverse outcomes may result despite adherence to best surgical practices.


A new approach to extending the wheat marker pool by anchored PCR amplification of compound SSRs.
A study was undertaken to determine the utility in bread wheat of anchored PCR for the development of single locus SSR markers targeted at compound repeat motifs. In anchored PCR, microsatellite amplification is achieved using a single primer complementary to the flanking sequence, and one which anchors to the repeat junction of the compound SSR. The recovery rate of useable markers was found to be similar (43%) to that reported for conventionally generated SSRs. Thus, anchored PCR can be used to reduce the costs of marker development, since it requires that only half the number of primers be synthesised. Where fluorescence-based platforms are used, marker deployment costs are lower, since only the anchoring primers need to be labelled. In addition, anchored PCR improves the recovery of useful markers, as it allows assays to be generated from microsatellite clones with repeat sequences located close to their ends, a situation where conventional PCR amplification fails as two flanking primers cannot be designed. Strategies to permit the large-scale development of compound SSR markers amplified by anchored PCR are discussed.


Investigation of an outbreak of acute respiratory disease in côte d'ivoire in april 2007.
This study aim was to investigate an outbreak of human cases of unexplained influenza-like illness and fatal acute respiratory infection (ARI), with simultaneous poultry illness and high mortality raising concerns of possible influenza A (H5N1), virus in Cote d'Ivoire in February and March 2007. To investigate the outbreak, we conducted active surveillance in the community and reviewed health registries. Persons meeting the case definition were asked to provide nasopharyngeal specimens. On the basis of clinical and epidemiological information, specimens were tested using conventional RT-PCR for the M gene of the influenza viruses and hemagglutinin H5 of avian influenza A (H5N1), virus; negative samples were tested for other respiratory viruses. Specimens from healthy animals were also collected. Between October 2006, and February 2007, 104 suspected cases of Acute Respiratory Disease that included; 31 deaths recorded. We collected and tested 73 nasopharyngeal specimens; of which, 2, were positive for human Coronavirus OC43 and 1 for influenza C virus. No pathogens were identified in animal specimens. The investigation quickly ruled out influenza A (H5N1), virus as the cause and found laboratory-confirmed cases of influenza C virus and human Coronavirus OC 43 for the first time in both Côte d'Ivoire and in a Sub-Saharan African country. However we were not able to show that these viruses caused the outbreak. Monitoring of influenza viruses must be a priority but other respiratory viruses and non-viral causes may be of interest too.


Liver fibrosis, cirrhosis, and cirrhosis-related nodules: Imaging diagnosis and surveillance.
Although biological scores and elastography continue to yield the best results, imaging retains a crucial role in the diagnosis of liver fibrosis and cirrhosis. First, digestive symptoms or biological liver test abnormalities often lead the referring physician to request an abdominal ultrasound, and with an experienced operator, accuracy of ultrasound can reach 85% for the diagnosis of severe fibrosis or cirrhosis. Second, imaging could lead to discovery of nonsymptomatic fibrosis or cirrhosis, with an estimated prevalence of 0.5-2.8% in the population. After diagnosis, imaging is central in the follow-up of cirrhosis. It is used to detect worsening of portal hypertension and hepatocellular carcinoma (HCC). Because many nodules are present in a cirrhotic liver, familiarity with the features of HCC can facilitate noninvasive diagnosis and early and accurate treatment.


Characterization of uterine leukocyte infiltration in gilts after artificial insemination.
The objective of this study was to characterize the uterine leukocyte influx after artificial insemination (AI). After detection of oestrus with a boar at intervals of 1.5 h, seventy-two gilts were randomly assigned to a 2 x 3 x 4 factorial arrangement. AI was performed with 100 ml extended semen containing 5 x 10(9) spermatozoa (semen; n = 36) or 100 ml VSP semen extender (extender; n = 36) at one of three times after detection of oestrus: 12, 24 or 36 h (n = 24/time). The uterus was lavaged at 6, 12, 18 or 24 h (n = 18/time) after AI to determine the total number of uterine leukocytes. In addition, uterine lavage was performed on nine untreated gilts immediately after the detection of oestrus to establish a baseline number of leukocytes. The leukocyte response in all samples consisted predominately (92-99%) of polymorphonuclear neutrophilic granulocytes (PMNs). The mean number of PMNs recovered from the uteri of gilts treated with semen was greater than in gilts treated with extender and in untreated gilts (P < 0.01). The greatest number of PMNs in semen-treated gilts was found 12 h after AI (P < 0.01), and this number was sustained for 24 h. In contrast, the number of uterine PMNs recovered from extender-treated gilts reached a peak at 6 h and had declined by 12 h after AI (P < 0.05). It was concluded that an extensive influx of PMNs into the uterus is a normal sequence to AI. The consequences and importance of semen-induced uterine leukocytosis needs further investigation.


Fournier's Gangrene: 10-Year Experience of a Plastic Surgery and Burns Department at a Tertiary Hospital.
Fournier gangrene is a polymicrobial life threatening infection of perineal subcutaneous soft tissues with its point of origin in urologic, colorectal or skin diseases. Although more frequent in elderly and men, it can affect all genders and age groups. Perianal abscess, diabetes mellitus and Escherichia coli are the most frequent cause, predisposing comorbidity, and microorganism found in tissue culture analysis respectively. The objective of this study was to describe the experience of a Plastic Surgery Department of a tertiary Hospital in reconstructing Fournier's gangrene perineal defects and its detailed demography. The sample is composed of all patients with Fournier gangrene admitted in the Plastic Surgery and Burns Department. The authors retrospectively collected and analyzed demographic and clinical data during a period of 10 years including gender, age, length of stay, cause, number of debridements, predisposing factors, microbial culture results, surgical reconstructive techniques and its associated complications, additional surgical procedures and outcomes. Fifteen patients were identified: 14 males (93%) and one female (7%); mean age was 66.9 years (range: 46 - 86); mean, length of stay was 46.8 days (range: 20 - 71 days) and mean number of debridements was 3.3 (range: 1 - 4). The most frequent predisposing factor was diabetes mellitus, the major cause was perianal (n = 2) and skin abscess (n = 2). Eight (53.3%) patients had no identifiable source of Fournier gangrene. Various types of reconstructive techniques were employed; and 5 additional surgical interventions (33.3%) were undertaken (one cystostomy, two orchidectomy, two ileostomy); six patients (40%) presented reconstructive technique complications with adequate final outcome. In contrast with the literature, where Escherichia coli was the most frequently isolated agent, Staphylococcus aureus was the most frequent microorganism found in tissue biopsy/pus collection analysis. A higher than expected number of patients (n = 8) had no identifiable source of Fournier gangrene. This findings can be explained by the retrospective non-multicentre study limitation, with a potencial source of bias patients that were transferred from other hospitals in advanced stage, without point of origin of Fournier's gangrene identified. Early recognition and extensive necrotic tissue debridement, along with prompt and adequate antimicrobial treatment, are the mainstay of Fournier gangrene management, thus reducing morbidity and mortality in these patients. Surgical reconstruction challenges derived from this condition should be addressed by specialized teams due to the risk of dysfunctional sequelae and conspicuous deformities. Taking in account the single-center and retrospective observational character of the present study, these premises require proper validation from a multicenter prospective study.


The importance of biomass net uptake for a trace metal budget in a forest stand in north-eastern France.
The trace metal (TM: Cd, Cu, Ni, Pb and Zn) budget (stocks and annual fluxes) was evaluated in a forest stand (silver fir, Abies alba Miller) in north-eastern France. Trace metal concentrations were measured in different tree compartments in order to assess TM partitioning and dynamics in the trees. Inputs included bulk deposition, estimated dry deposition and weathering. Outputs were leaching and biomass exportation. Atmospheric deposition was the main input flux. The estimated dry deposition accounted for about 40% of the total trace metal deposition. The relative importance of leaching (estimated by a lumped parameter water balance model, BILJOU) and net biomass uptake (harvesting) for ecosystem exportation depended on the element. Trace metal distribution between tree compartments (stem wood and bark, branches and needles) indicated that Pb was mainly stored in the stem, whereas Zn and Ni, and to a lesser extent Cd and Cu, were translocated to aerial parts of the trees and cycled in the ecosystem. For Zn and Ni, leaching was the main output flux (>95% of the total output) and the plot budget (input-output) was negative, whereas for Pb the biomass net exportation represented 60% of the outputs and the budget was balanced. Cadmium and Cu had intermediate behaviours, with 18% and 30% of the total output relative to biomass exportation, respectively, and the budgets were negative. The net uptake by biomass was particularly important for Pb budgets, less so for Cd and Cu and not very important for Zn and Ni in such forest stands.


Parkinson-like features in ALS with predominant upper motor neuron involvement.
Owing to the frequent observation of poverty of movements, facial hypomimia and balance impairment, amyotrophic lateral sclerosis (ALS) variant with predominance of upper motor neuron involvement (UMN-ALS) is prone to be diagnosed with Parkinsonism. A clinical assessment, including the velocity-dependent stretch response test to differentiate between pyramidal and extrapyramidal stiffness; the Unified Parkinson's Disease Rating Scale and the Berg Balance Scale to assess degree of bradykinesia and postural instability; and (123)I-FP-CIT scintigraphy evaluation to investigate the nigrostriatal circuit involvement, were carried out to characterize Parkinson-like features in UMN-ALS patients. Sixteen UMN-ALS patients were included in the study. The velocity-dependent stretch response indicated spasticity in all the muscles tested. The degree of stiffness was found to be related to bradykinesia and postural instability. Eleven patients (70%) showed a reduction in striatal (123)I-FP-CIT uptake found to be related to disease duration and patients' ages but not to scores of the functional scales. Slowness of movements and postural instability noted in our patients could be mostly attributed to spasticity. The lack of any correlation between UPDRS or BBS scores and the degree of nigrostriatal impairment on DaTSCAN seems to disprove nigrostriatal circuit involvement in these extrapyramidal-like features.


State carnivals and the subvention of multiculturalism in Singapore.
Increasing attention is being paid to the specificities of Asian multiculturalism in relation to ethnic pluralism, citizenship and developmental state formation. This article examines these relationships by analysing three carnival events in colonial and postcolonial Singapore that were organized by the state to promote its official multiculturalism. Through its cultural logics of horizontal racial segmentation, cascading symbolic authority from the state to co-opted communal representatives and multi-modal ritual iteration, the 1937 King George VI coronation celebrations proffered an imperial multiculturalism based on mediating plural groups and procedural norms. Adopting the same cultural logics in the 1970s, the newly-independent nation-state revived and transformed Chingay, a creole Chinese religious procession, into an annual parade celebrating the nation as comprising racially plural groups bound together by the modern ethos of progress the developmental state exemplified. In the 2000s, Chingay has been turned into an international spectacle celebrating Singapore as a cosmopolitan global city of hybridizing multiculturalism. But indicative of new racial-class segmentation, the old nation-building pluralism is promoted by Racial Harmony Day carnivals held in suburban public housing neighborhoods. This bifurcated multiculturalism reflects the developmental state's attempts to deal with new citizenship trends as they grind against the old ethnic pluralism. While faced with the same issues globalization brings, this postcolonial multiculturalism is distinctively different from liberal multiculturalism, not least because the subvention of multiculturalism is achieved through the state appropriation of vernacular cultural practices through its carnivals.


A new approach to neuroimaging with magnetoencephalography.
We discuss the application of beamforming techniques to the field of magnetoencephalography (MEG). We argue that beamformers have given us an insight into the dynamics of oscillatory changes across the cortex not explored previously with traditional analysis techniques that rely on averaged evoked responses. We review several experiments that have used beamformers, with special emphasis on those in which the results have been compared to those observed in functional magnetic resonance imaging (fMRI) and on those studying induced phenomena. We suggest that the success of the beamformer technique, despite the assumption that there are no linear interactions between the mesoscopic local field potentials across distinct cortical areas, may tell us something of the balance between functional integration and segregation in the human brain. What is more, MEG beamformer analysis facilitates the study of these complex interactions within cortical networks that are involved in both sensory-motor and cognitive processes.


Light-dependent pathways for dopaminergic amacrine cell development and function.
Retinal dopamine is a critical modulator of high acuity, light-adapted vision and photoreceptor coupling in the retina. Dopaminergic amacrine cells (DACs) serve as the sole source of retinal dopamine, and dopamine release in the retina follows a circadian rhythm and is modulated by light exposure. However, the retinal circuits through which light influences the development and function of DACs are still unknown. Intrinsically photosensitive retinal ganglion cells (ipRGCs) have emerged as a prime target for influencing retinal dopamine levels because they costratify with DACs in the inner plexiform layer and signal to them in a retrograde manner. Surprisingly, using genetic mouse models lacking specific phototransduction pathways, we find that while light influences the total number of DACs and retinal dopamine levels, this effect does not require ipRGCs. Instead, we find that the rod pathway is a critical modulator of both DAC number and retinal dopamine levels.


Communicating uncertain experimental evidence.
Four experiments examined when laypeople attribute unexpected experimental outcomes to error, in foresight and in hindsight, along with their judgments of whether the data should be published. Participants read vignettes describing hypothetical experiments, along with the result of the initial observation, considered as either a possibility (foresight) or a reality (hindsight). Experiment 1 found that the initial observation seemed more likely to be replicated when viewed in hindsight than in foresight. The remaining experiments contrasted responses to an initial observation from 1 of the 4 studies that was either expected or unexpected (based on the predictions of participants in Experiment 1). Experiments 2A-C and Experiment 3 found that unexpected results were more likely to be attributed to methodological problems than were expected ones-but to the same degree in foresight and in hindsight. Participants in Experiment 4 had more confidence in an explanation for an unexpected outcome when it was mentioned before that outcome was revealed than when it was suggested only after the surprise was known. In all the experiments, most participants recommended collecting more data before publishing the results, especially when they attributed the results to multiple causes. The results suggest that considering the causes of unexpected experimental results in foresight may improve the evaluation and communication of those results in hindsight.


Selective association of the methyl-CpG binding protein MBD2 with the silent p14/p16 locus in human neoplasia.
DNA methylation of tumor suppressor genes is a common feature of human cancer. The cyclin-dependent kinase inhibitor gene p16/Ink4A is hypermethylated in a wide range of malignant tissues and the p14/ARF gene located 20 kb upstream on chromosome 9p21 is also methylated in carcinomas. p14/ARF (ARF, alternative reading frame) does not inhibit the activities of cyclins or cyclin-dependent kinase complexes; however, the importance of the two gene products in the etiology of cancer resides in their involvement in two major cell cycle regulatory pathways: p53 and the retinoblastoma protein, Rb, respectively. Distinct first exons driven from separate promoters are spliced onto the common exons 2 and 3 and the resulting proteins are translated in different reading frames. Both genes are expressed in normal cells but can be alternatively or coordinately silenced when their CpG islands are hypermethylated. Herein, we examined the presence of methyl-CpG binding proteins associated with aberrantly methylated promoters, the distribution of acetylated histones H3 and H4 by chromatin immunoprecipitation assays, and the effect of chemical treatment with 5-aza-2'-deoxycytidine (5aza-dC) and trichostatin A on gene induction in colon cell lines by quantitative reverse transcriptase-PCR. We observed that the methyl-CpG binding protein MBD2 is targeted to methylated regulatory regions and excludes the acetylated histones H3 and H4, resulting in a localized inactive chromatin configuration. When methylated, the genes can be induced by 5aza-dC but the combined action of 5aza-dC and trichostatin A results in robust gene expression. Thus, methyl-CpG binding proteins and histone deacetylases appear to cooperate in vivo, with a dominant effect of DNA methylation toward histone acetylation, and repress expression of tumor suppressor genes hypermethylated in cancers.


Proteomic and Transcriptomic Patterns during Lipid Remodeling in Nannochloropsis gaditana.
Nutrient limited conditions are common in natural phytoplankton communities and are often used to increase the yield of lipids from industrial microalgae cultivations. Here we studied the effects of bioavailable nitrogen (N) and phosphorus (P) deprivation on the proteome and transcriptome of the oleaginous marine microalga Nannochloropsis gaditana. Turbidostat cultures were used to selectively apply either N or P deprivation, controlling for variables including the light intensity. Global (cell-wide) changes in the proteome were measured using Tandem Mass Tag (TMT) and LC-MS/MS, whilst gene transcript expression of the same samples was quantified by Illumina RNA-sequencing. We detected 3423 proteins, where 1543 and 113 proteins showed significant changes in abundance in N and P treatments, respectively. The analysis includes the global correlation between proteomic and transcriptomic data, the regulation of subcellular proteomes in different compartments, gene/protein functional groups, and metabolic pathways. The results show that triacylglycerol (TAG) accumulation under nitrogen deprivation was associated with substantial downregulation of protein synthesis and photosynthetic activity. Oil accumulation was also accompanied by a diverse set of responses including the upregulation of diacylglycerol acyltransferase (DGAT), lipase, and lipid body associated proteins. Deprivation of phosphorus had comparatively fewer, weaker effects, some of which were linked to the remodeling of respiratory metabolism.


External validation of the preoperative anatomical classification for prediction of complications related to nephron-sparing surgery.
Ficarra et al. (Eur Urol 56:786-793, 2009) published a preoperative anatomical classification (PADUA) to assess the impact of anatomical parameters of renal tumors on complication rate of nephron-sparing surgery (NSS). The objective of this study is to provide a bi-center external validation of this classification using the technique of hilar arterial clamping during open and laparoscopic NSS and to correlate the PADUA score to the ischemia time. 240 consecutive tumors treated with open and laparoscopic NSS were reclassified according to the PADUA classification. Complications were graded according to the modified Clavien system (Dindo et al. in Ann Surg 240:205-213, 2004). Chi-square tests and multivariate logistic regression models addressed the predictive value of PADUA classification on overall complication rate and grade. Mean patient age was 62.2 +/- 13.3 years. Eastern Cooperative Oncology Group performance was 0 in 76%, 1 in 22% and 2 in 2%. 61 (25%) were treated laparoscopically. The median PADUA score was 7.5 (range 6-13). Mean surgery and ischemia time was 189 +/- 95 and 24 +/- 22 min, respectively. Overall complication rate was 23% (n = 54). On univariate analysis, the PADUA score correlated with complication rate (p < 0.001) of open and laparoscopic NSS. On multivariate, only the PADUA score correlated with complication rate (p = 0.0056). Ischemic time correlated with the PADUA score and was significantly higher in PADUA score > or = 10 (p = 0.034). The PADUA score is a reliable tool to preoperatively predict the risk of complications. In addition, it might be beneficial for a more objective patient selection for laparoscopic surgery and teaching NSS.


Sesamol exhibits potent antimycobacterial activity: Underlying mechanisms and impact on virulence traits.
Novel strategies to overcome multidrug resistance (MDR) in Tuberculosis (TB) still remain a concern. Usage of natural compounds nowadays to surmount the increasing burden of MDR-TB has shown promising results. The aim of this study was to evaluate the antimycobacterial potential of sesamol (Ses) a natural phenolic compound against Mycobacterium smegmatis, a surrogate for MTB and its underlying mechanism of action along with its effect on mycobacterial virulence traits. Cell surface phenotypes were estimated microscopically and spectrophotometrically respectively. Membrane parameters were assessed using propidium iodide (PI) uptake, passive diffusion of drug with substrate EtBr and phenotypic susceptibility assay. Changes in lipid profiles were estimated by lipase assay. Oxidative and genotoxic damage were studied using fluorescent probes DCFDA and DAPI. Biofilm formation was studied using crystal violet and calcoflour white staining probes along with biomass measurement. Cell adherence was estimated using buccal epithelial cells. We observed that antimycobacterial activity of Ses was 6mM and it enhances the efficiency of known anti-TB drugs. Ses affects cell surface phenotypes as displayed by altered colony morphology, impaired sliding motility and enhanced cell sedimentation rate. Membrane perturbation was revealed by hypersensitivity against SDS, reduced PI uptake, enhanced passive diffusion and lipase activity. In addition, Ses leads to oxidative and DNA damage along with abrogated iron homeostasis. Furthermore, we uncover phenotypes related to virulence like inhibited biofilm formation and cell adherence to buccal epithelial cells. This study for the first time establishes the anti-mycobacterial potential of Ses that may be further exploited for improving the therapeutic strategies and warrants further attention.


Gbx2, a LIF/Stat3 target, promotes reprogramming to and retention of the pluripotent ground state.
Activation of signal transducer and activator of transcription 3 (Stat3) by leukemia inhibitory factor (LIF) maintains mouse embryonic stem cell (mESC) self-renewal and also facilitates reprogramming to ground state pluripotency. Exactly how LIF/Stat3 signaling exerts these effects, however, remains elusive. We identified gastrulation brain homeobox 2 (Gbx2) as a LIF/Stat3 downstream target that, when overexpressed, allows long-term expansion of undifferentiated mESCs in the absence of LIF/Stat3 signaling. Elevated Gbx2 expression also enhanced reprogramming of mouse embryonic fibroblasts to induced pluripotent stem cells. Moreover, overexpression of Gbx2 was sufficient to reprogram epiblast stem cells to ground state ESCs. Our results reveal a novel function of Gbx2 in mESC reprogramming and LIF/Stat3-mediated self-renewal.


Controversies in drug allergy: Testing for delayed reactions.
Controversies exist with regard to in vivo approaches to delayed immunologically mediated adverse drug reactions, such as exanthem (maculopapular eruption), drug reaction with eosinophilia and systemic symptoms, acute generalized exanthematous pustulosis, Stevens-Johnson syndrome/toxic epidermal necrolysis, and fixed drug eruptions. In particular, widespread differences exist between regions and practice on the availability and use of intradermal and patch testing, the standard drug concentrations used, the use of additional drugs in intradermal and patch testing to help determine cross-reactivity, the timing of testing in relation to the occurrence of the adverse drug reaction, the use of testing in specific phenotypes, and the use of oral challenge in conjunction with delayed intradermal and patch testing to ascertain drug tolerance. It was noted that there have been advances in the science of delayed T cell-mediated reactions that have shed light on immunopathogenesis and provided a mechanism of preprescription screening in the case of HLA-B*57:01 and abacavir hypersensitivity and HLA-B*15:02 and carbamazepine Stevens-Johnson syndrome/toxic epidermal necrolysis in Southeast Asian subjects. Future directions should include the collaboration of large international networks to develop and standardize in vivo diagnostic approaches, such as skin testing and patch testing, combined with ex vivo and in vitro laboratory approaches.


Safety, Efficacy and Long-Term Outcomes of Patients Treated with the Occlutech Paravalvular Leak Device for Significant Paravalvular Regurgitation.
Between December 2014 and March 2021, 144 patients with aortic (Ao) or mitral (Mi) paravalvular leaks (PVLs) were enrolled at 21 sites in 10 countries. Safety data were available for 137 patients, who were included in the safety analysis fraction (SAF), 93 patients with Mi PVLs and 44 patients with Ao PVLs. The full analysis set (FAS) comprised 112 patients with available stratum (aortic/mitral leak) as well as baseline (BL), 180-day or later assessments (2 years). Procedural success (implantation of the device with a proper closure of the PVL, defined as reduction in paravalvular regurgitation of ≥one grade as assessed by echocardiography post implantation) was achieved in 91.3% of FAS patients with Mi PVLs and in 90.0% of those with Ao PVLs. The proportion of patients suffering from significant or severe heart failure (HF), classified as New York Heart Association (NYHA) class III/IV, decreased from 80% at baseline to 14.1% at 2-year follow-up (FAS). The proportion of FAS patients needing hemolysis-related blood transfusion decreased from 35.5% to 3.8% and from 8.1% to 0% in Mi patients and Ao patients, respectively. In total, 35 serious adverse events (SAEs) were reported in 27 patients (19.7%) of the SAF population. The SAEs considered possibly or probably related to the device included device embolization (three patients), residual leak (two patients) and vascular complication (one patient). During follow-up, 12/137 (8.8%) patients died, but none of the deaths was considered to be device-related. Patients implanted with the Occlutech Paravalvular Leak Device (PLD) showed long-lasting improvements in clinical parameters, including NYHA class and a reduced dependency on hemolysis-related blood transfusions.


Synthesis, characterization, and bioavailability in rats of ferric phosphate nanoparticles.
Particle size is a determinant of iron (Fe) absorption from poorly soluble Fe compounds. Decreasing the particle size of metallic Fe and ferric pyrophosphate added to foods increases Fe absorption. The aim of this study was to develop and characterize nanoparticles of FePO(4) and determine their bioavailability and potential toxicity in rats. Amorphous FePO(4) nanopowders with spherical structure were synthesized by flame spray pyrolysis (FSP). The nanopowders were characterized and compared with commercially available FePO(4) and FeSO(4), including measurements of specific surface area (SSA), structure by transmission electron microscopy, in vitro solubility at pH 1 and 2, and relative bioavailability value (RBV) to FeSO(4) in rats using the hemoglobin repletion method. In the latter, the potential toxicity after Fe repletion was assessed by histological examination and measurement of thiobarbituric acid reactive substances (TBARS). The commercial FePO(4) and the 2 FePO(4) produced by FSP (mean particle sizes, 30.5 and 10.7 nm) had the following characteristics: SSA: 32.6, 68.6, 194.7 m(2)/g; in vitro solubility after 30 min at pH 1: 73, 79, and 85% of FeSO(4); and RBV: 61, 70, and 96%, respectively. In the histological examinations and TBARS analysis, there were no indications of toxicity. In conclusion, nanoparticles of FePO(4) have a solubility and RBV not significantly different from FeSO(4). Reducing poorly soluble Fe compounds to nanoscale may increase their value for human nutrition.


Formulation, release characteristics, and bioavailability study of gastroretentive floating matrix tablet and floating raft system of Mebeverine HCl.
To prolong the residence time of dosage forms within the gastrointestinal tract until all drug is released at the desired rate is one of the real challenges for oral controlled-release drug delivery systems. This study was designed to develop a controlled-release floating matrix tablet and floating raft system of Mebeverine HCl (MbH) and evaluate different excipients for their floating behavior and in vitro controlled-release profiles. Oral pharmacokinetics of the optimum matrix tablet, raft system formula, and marketed Duspatalin® 200 mg retard as reference were studied in beagle dogs. The optimized tablet formula (FT-10) and raft system formula (FRS-11) were found to float within 34±5 sec and 15±7 sec, respectively, and both remain buoyant over a period of 12 h in simulated gastric fluid. FT-10 (Compritol/HPMC K100M 1:1) showed the slowest drug release among all prepared tablet formulations, releasing about 80.2% of MbH over 8 h. In contrast, FRS-11 (Sodium alginate 3%/HPMC K100M 1%/Precirol 2%) had the greatest retardation, providing sustained release of 82.1% within 8 h. Compared with the marketed MbH product, the Cmax of FT-10 was almost the same, while FRS-11 maximum concentration was higher. The tmax was 3.33, 2.167, and 3.0 h for marketed MbH product, FT-10, and FRS-11, respectively. In addition, the oral bioavailability experiment showed that the relative bioavailability of the MbH was 104.76 and 116.01% after oral administration of FT-10 and FRS-11, respectively, compared to marketed product. These results demonstrated that both controlled-released floating matrix tablet and raft system would be promising gastroretentive delivery systems for prolonging drug action.


The lost children.
Women who have lost children to perinatal complications, are subjected to pain and grief continuously; however their agony increases on remembrance days such as birthdate, or on Mother's Day. Fathers, siblings and grandparents suffer too. Common disorders of pregnancy, such as pregnancy-induced hypertension (PIH), or the more serious pre-eclampsia, HELLP syndrome, or eclampsia, can lead to devastating effects such as miscarriage, stillbirth, or neonatal death; or at the very least, a sick infant. With many of these consequences, the loss of the dreams, hopes and plans that parents have made is imminent. The investigation of the psychosocial aspects of 'high-risk' pregnancy has never been fully addressed. However, the threat of loss, or the actual experience, may provoke the onset of a potential psychological crisis during the perinatal period. Therefore, it is important that these issues be addressed by the nurse in order to aid the development of coping mechanisms to enable women and their families to deal with what may happen. This may be done by predicting the stages of the bereavement process experienced by these women and their family members, as outlined in the Kubler-Ross model of bereavement (1969), which is indicative of many types of grief reactions. Other issues including the restriction in activity, uncertainty of pregnancy outcomes, disruption in work or career activities, financial strains, and reduced labour and birthing options, become concerns for high-risk pregnant women. The way women deal with these issues and the pathways nurses can take to help these women develop effective coping strategies, will be addressed also.


Health technology assessment of paediatric medicines: European landscape, challenges and opportunities inside the conect4children project.
The medicine development process is complex and requires time and effort to ensure safety, efficacy and quality. In paediatrics, this process is even more challenging, as it involves a subgroup of the population that already faces a considerable gap in the clinical evaluation of medicines and devices compared to the adult population. Moreover, access to therapies is heavily influenced by national health technology assessment (HTA) recommendations, which often form the basis for pricing and reimbursement decisions that affect the availability of effective treatments within the national health systems. Yet performing an HTA to assess the relative effectiveness and cost-effectiveness of a new children's treatment has several non-trivial implications, creating a critical issue for the paediatric population. In addition, the advent of innovative health technologies for children emphasises the need to empower the role of HTAs in paediatrics. This article aims at describing the most relevant elements of the drug development process in the paediatric field by focusing on the HTA. Particular attention will be paid to the factors that influence market access for new paediatric medicines and patients' access to treatment. The article will also highlight some central methodological challenges in conducting HTA in the paediatric field. Finally, the article will provide insight into how initiatives, such as conect4children, may subsequently reinforce HTA awareness in the paediatric community and strengthen collaborations through network mechanisms.


A two-component regulatory system interconnects resistance to polymyxins, aminoglycosides, fluoroquinolones, and β-lactams in Pseudomonas aeruginosa.
Constitutive overexpression of the active efflux system MexXY/OprM is a major cause of resistance to aminoglycosides, fluoroquinolones, and cefepime in clinical strains of Pseudomonas aeruginosa. Upregulation of this pump often results from mutations occurring in mexZ, the local repressor gene of the mexXY operon. In this study, analysis of MexXY-overproducing mutants selected in vitro from reference strain PAO1Bes on amikacin (at a concentration 1.5-fold higher than the MIC) led to identification of a new class of mutants harboring an intact mexZ gene and exhibiting increased resistance to colistin and imipenem in addition to aminoglycosides, fluoroquinolones, and cefepime. Reverse transcription-quantitative PCR (RT-qPCR) experiments on a selected clone named PAOW2 demonstrated that mexXY overexpression was independent of mexZ and the PA5471 gene, which is required for drug-dependent induction of mexXY. Furthermore, the transcript levels of the oprD gene, which encodes the carbapenem-selective porin OprD, were found to be reduced drastically in PAOW2. Whole-genome sequencing revealed a single mutation resulting in an M59I substitution in the ParR protein, the response regulator of the ParRS two-component regulatory system (with ParS being the sensor kinase), which is required for adaptive resistance of P. aeruginosa to polycationic peptides such as colistin. The multidrug resistance phenotype was suppressed in PAOW2 by deletion of the parS and parRS genes and conferred to PAO1Bes by chromosomal insertion of the mutated parRS locus from PAOW2. As shown by transcriptomic analysis, only a very small number of genes were expressed differentially between PAOW2 and PAO1Bes, including the lipopolysaccharide (LPS) modification operon arnBCADTEF-ugd, responsible for resistance to polycationic agents. Exposure of wild-type PAO1Bes to different polycationic peptides, including colistin, was shown to result in increased mexY and repressed oprD expression via ParRS, independent of PA5471. In agreement with these results, colistin antagonized activity of the MexXY/OprM substrates in PAO1Bes but not in a ΔparRS derivative. Finally, screening of clinical strains exhibiting the PAOW2 resistance phenotype allowed the identification of additional alterations in ParRS. Collectively, our data indicate that ParRS may promote either induced or constitutive multidrug resistance to four different classes of antibiotics through the activation of three distinct mechanisms (efflux, porin loss, and LPS modification).


Murine cytomegalovirus infection exacerbates complex IV deficiency in a model of mitochondrial disease.
The influence of environmental insults on the onset and progression of mitochondrial diseases is unknown. To evaluate the effects of infection on mitochondrial disease we used a mouse model of Leigh Syndrome, where a missense mutation in the Taco1 gene results in the loss of the translation activator of cytochrome c oxidase subunit I (TACO1) protein. The mutation leads to an isolated complex IV deficiency that mimics the disease pathology observed in human patients with TACO1 mutations. We infected Taco1 mutant and wild-type mice with a murine cytomegalovirus and show that a common viral infection exacerbates the complex IV deficiency in a tissue-specific manner. We identified changes in neuromuscular morphology and tissue-specific regulation of the mammalian target of rapamycin pathway in response to viral infection. Taken together, we report for the first time that a common stress condition, such as viral infection, can exacerbate mitochondrial dysfunction in a genetic model of mitochondrial disease.


Gemini surfactants mediate efficient mitochondrial gene delivery and expression.
Gene delivery targeting mitochondria has the potential to transform the therapeutic landscape of mitochondrial genetic diseases. Taking advantage of the nonuniversal genetic code used by mitochondria, a plasmid DNA construct able to be specifically expressed in these organelles was designed by including a codon, which codes for an amino acid only if read by the mitochondrial ribosomes. In the present work, gemini surfactants were shown to successfully deliver plasmid DNA to mitochondria. Gemini surfactant-based DNA complexes were taken up by cells through a variety of routes, including endocytic pathways, and showed propensity for inducing membrane destabilization under acidic conditions, thus facilitating cytoplasmic release of DNA. Furthermore, the complexes interacted extensively with lipid membrane models mimicking the composition of the mitochondrial membrane, which predicts a favored interaction of the complexes with mitochondria in the intracellular environment. This work unravels new possibilities for gene therapy toward mitochondrial diseases.