The UNESCO Universal Declaration on Bioethics and Human Rights: perspectives from Kenya and South Africa.
In October 2005, UNESCO (the United Nations Educational, Scientific and Cultural Organization) adopted the Universal Declaration on Bioethics and Human Rights. This was the culmination of nearly 2 years of deliberations and negotiations. As a non-binding instrument, the declaration must be incorporated by UNESCO's member states into their national laws, regulations or policies in order to take effect. Based on documentary evidence and data from interviews, this paper compares the declaration's universal principles with national bioethics guidelines and practice in Kenya and South Africa. It concentrates on areas of particular relevance to developing countries, such as protection of vulnerable persons and social responsibility. The comparison demonstrates the need for universal principles to be contextualised before they can be applied in a meaningful sense at national level. The paper also assesses the 'added value' of the declaration in terms of biomedical research ethics, given that there are already well-established international instruments on bioethics, namely the World Medical Association Declaration of Helsinki and the CIOMS (Council for International Organizations of Medical Sciences) guidelines on biomedical research. It may be that the added value lies as much in the follow-up capacity building activities being initiated by UNESCO as in the document itself.


Incremental lines of von Ebner in dinosaurs and the assessment of tooth replacement rates using growth line counts.
Dinosaur dentine exhibits growth lines that are tens of micrometers in width. These laminations are homologous to incremental lines of von Ebner found in extant mammal and crocodilian teeth (i.e., those of amniotes). The lines likely reflect daily dentine formation, and they were used to infer tooth development and replacement rates. In general, dinosaur tooth formation rates negatively correlated with tooth size. Theropod tooth replacement rates negatively correlated with tooth size, which was due to limitations in the dentine formation rates of their odontoblasts. Derived ceratopsian and hadrosaurian dinosaurs retained relatively rapid tooth replacement rates through ontogeny. The evolution of dental batteries in hadrosaurs and ceratopsians can be explained by dentine formation constraints and rapid tooth wear. In combination with counts of shed dinosaur teeth, tooth replacement rate data can be used to assess population demographics of Mesozoic ecosystems. Finally, it is of historic importance to note that Richard Owen appears to have been the first to observe incremental lines of von Ebner in dinosaurs more than 150 years ago.


Antagonism of xylazine-pentobarbital anesthesia by yohimbine in ponies.
Effects of yohimbine on xylazine-pentobarbital anesthesia were evaluated in ponies. Five minutes after the IV injection of xylazine (1.1 mg/kg of body weight), pentobarbital sodium (12.7 mg/kg, IV) and additional xylazine (2.2 mg/kg, IM) were given and produced anesthesia in 12 ponies for 64.0 +/- 16.4 minutes (mean +/- SD) as well as immobilization for 89.8 +/- 34.2 minutes. Eleven ponies were given yohimbine (0.1 mg/kg, IV) 50 minutes after pentobarbital dosing. In these 11 ponies, durations of anesthesia and immobilization were shorter, 52.0 +/- 1.4 and 65.5 +/- 14.8 minutes, respectively. The xylazine-pentobarbital combination caused bradycardia that was reversed by yohimbine injection. Xylazine-pentobarbital produced a small, but steady, decrease of mean arterial blood pressure, which was compounded by yohimbine administration and was evident for approximately 2 minutes. Within a minute after yohimbine injection, the ponies' respiratory rate decreased and the length of inspiration and expiration and thoracic breathing increased. This lasted approximately 2 to 3 minutes and was followed by an increase in respiratory rate. The anesthesia also produced a decrease in PaO2 that gradually returned to base line in 12 control ponies, but was more pronounced in 11 ponies given yohimbine. The PaCO2, although remaining moderately high in control ponies, returned to base line after yohimbine injection. An increased pHa was seen 60 minutes after induction of anesthesia and was especially noticeable after yohimbine administration. Decreases in the number of WBC, hemoglobin content, PCV, plasma protein and serum aspartate transaminase resulting from xylazine-pentobarbital were reversed by yohimbine. Conversely, serum glucose values and creatine kinase activities were increased by xylazine-pentobarbital.(ABSTRACT TRUNCATED AT 250 WORDS)


The many facets of H/ACA ribonucleoproteins.
The H/ACA ribonucleoproteins (RNPs) are known as one of the two major classes of small nucleolar RNPs. They predominantly guide the site-directed pseudouridylation of target RNAs, such as ribosomal and spliceosomal small nuclear RNAs. In addition, they process ribosomal RNA and stabilize vertebrate telomerase RNA. Taken together, the function of H/ACA RNPs is essential for ribosome biogenesis, pre-mRNA splicing, and telomere maintenance. Every cell contains 100-200 different species of H/ACA RNPs, each consisting of the same four core proteins and one function-specifying H/ACA RNA. Most of these RNPs reside in nucleoli and Cajal bodies and mediate the isomerization of specific uridines to pseudouridines. Catalysis of the reaction is mediated by the putative pseudouridylase NAP57 (dyskerin, Cbf5p). Unexpectedly, mutations in this housekeeping enzyme are the major determinants of the inherited bone marrow failure syndrome dyskeratosis congenita. This review details the many diverse functions of H/ACA RNPs, some yet to be uncovered, with an emphasis on the role of the RNP proteins. The multiple functions of H/ACA RNPs appear to be reflected in the complex phenotype of dyskeratosis congenita.


The Ameliorative Impact of Cichorium intybus L. Distillate on Reproductive Parameters in Male Mice.
Cichorium intybus L. (Kasni) distillate is widely used in Eastern countries as a safe herbal drink to improve male fertility. However, the potential effects on fertility parameters and possible adverse effects have not been studied experimentally. The current study aims to evaluate the impact of Cichorium intybus L. distillate (CD) on male mice fertility. In the present study (Shiraz, Iran), 30 male mice (30-35 g) were divided into three groups. 10 mice received distilled water (DW) for five weeks as the control group. Another 10 mice, named group CD1/2, received chicory distillate of 1/2 dilution, and the other 10 mice received chicory distillate of CD1/4 dilution as CD1/4 group, ad libitum for three weeks, and they received DW for two weeks afterward. Experimental mice were sacrificed on day 35, and sperm analysis and sera collection were performed for further investigation of FSH, LH, testosterone, and some liver and kidney function parameters. We used the left testis for stereological analysis, and the right one was excised to investigate the expression of the androgen receptor gene. For statistical analysis using SPSS 18.0, mean±SD values were analyzed by one-way analysis of variance (ANOVA) with Dunnett's analysis as post hoc to compare between groups. In stereological investigations, the Kruskal-Wallis method was used for pairwise comparisons to compare groups. The P value was considered statistically significant at P<0.05. Treatment with CD1/2 resulted in the elevation of serum FSH (P=0.002), LH (P=0.009), testosterone (P=0.034), seminiferous tubule epithelium volume (P=0.029) and length (P=0.028), and Leydig cells number (P=0.009) in comparison with the control group. Administrating CD1/2 (P=0.038) and CD1/4 (P=0.013) significantly increased sperm count compared to the control group. The results revealed that using chicory distillate can improve hormone levels and sperm count in male mice.


Applicability of computational systems biology in toxicology.
Systems biology as a research field has emerged within the last few decades. Systems biology, often defined as the antithesis of the reductionist approach, integrates information about individual components of a biological system. In integrative systems biology, large data sets from various sources and databases are used to model and predict effects of chemicals on, for instance, human health. In toxicology, computational systems biology enables identification of important pathways and molecules from large data sets; tasks that can be extremely laborious when performed by a classical literature search. However, computational systems biology offers more advantages than providing a high-throughput literature search; it may form the basis for establishment of hypotheses on potential links between environmental chemicals and human diseases, which would be very difficult to establish experimentally. This is possible due to the existence of comprehensive databases containing information on networks of human protein-protein interactions and protein-disease associations. Experimentally determined targets of the specific chemical of interest can be fed into these networks to obtain additional information that can be used to establish hypotheses on links between the chemical and human diseases. Such information can also be applied for designing more intelligent animal/cell experiments that can test the established hypotheses. Here, we describe how and why to apply an integrative systems biology method in the hypothesis-generating phase of toxicological research.


Pathogenic Variants in NUP214 Cause "Plugged" Nuclear Pore Channels and Acute Febrile Encephalopathy.
We report biallelic missense and frameshift pathogenic variants in the gene encoding human nucleoporin NUP214 causing acute febrile encephalopathy. Clinical symptoms include neurodevelopmental regression, seizures, myoclonic jerks, progressive microcephaly, and cerebellar atrophy. NUP214 and NUP88 protein levels were reduced in primary skin fibroblasts derived from affected individuals, while the total number and density of nuclear pore complexes remained normal. Nuclear transport assays exhibited defects in the classical protein import and mRNA export pathways in affected cells. Direct surface imaging of fibroblast nuclei by scanning electron microscopy revealed a large increase in the presence of central particles (known as "plugs") in the nuclear pore channels of affected cells. This observation suggests that large transport cargoes may be delayed in passage through the nuclear pore channel, affecting its selective barrier function. Exposure of fibroblasts from affected individuals to heat shock resulted in a marked delay in their stress response, followed by a surge in apoptotic cell death. This suggests a mechanistic link between decreased cell survival in cell culture and severe fever-induced brain damage in affected individuals. Our study provides evidence by direct imaging at the single nuclear pore level of functional changes linked to a human disease.


The prevention of gambling problems in youth: a conceptual framework.
Despite increased awareness of the need to begin educating young children about the potential dangers of gambling, empirical knowledge of the prevention of adolescent problem gambling and its translation into science-based prevention initiatives is scarce. This paper poses the question of whether or not the common elements of tobacco, alcohol, and illicit drug abuse prevention programs can be applied to gambling prevention. Common risk and protective factors across addictions, including gambling, appear to point to the need to develop a general model of primary, secondary, and tertiary prevention. The authors present the need for science-based prevention initiatives and describe a general adolescent risk-taking model as a basis for science-based prevention of adolescent problem gambling and other risk behaviors.


In Situ Growth of Halophilic Bacteria in Saline Fracture Fluids from 2.4 km below Surface in the Deep Canadian Shield.
Energy derived from water-rock interactions such as serpentinization and radiolysis, among others, can sustain microbial ecosystems deep within the continental crust, expanding the habitable biosphere kilometers below the earth's surface. Here, we describe a viable microbial community including sulfate-reducing microorganisms from one such subsurface lithoautotrophic ecosystem hosted in fracture waters in the Canadian Shield, 2.4 km below the surface in the Kidd Creek Observatory in Timmins, Ontario. The ancient groundwater housed in fractures in this system was previously shown to be rich in abiotically produced hydrogen, sulfate, methane, and short-chain hydrocarbons. We have further investigated this system by collecting filtered water samples and deploying sterile in situ biosampler units into boreholes to provide an attachment surface for the actively growing fraction of the microbial community. Scanning electron microscopy, energy-dispersive X-ray spectroscopy, and DNA sequencing analyses were undertaken to classify the recovered microorganisms. Moderately halophilic taxa (e.g., Marinobacter, Idiomarina, Chromohalobacter, Thiobacillus, Hyphomonas, Seohaeicola) were recovered from all sampled boreholes, and those boreholes that had previously been sealed to equilibrate with the fracture water contained taxa consistent with sulfate reduction (e.g., Desulfotomaculum) and hydrogen-driven homoacetogenesis (e.g., Fuchsiella). In contrast to this "corked" borehole that has been isolated from the mine environment for approximately 7 years at the time of sampling, we sampled additional open boreholes. The waters flowing freely from these open boreholes differ from those of the long-sealed borehole. This work complements ongoing efforts to describe the microbial diversity in fracture waters at Kidd Creek in order to better understand the processes shaping life in the deep terrestrial subsurface. In particular, this work demonstrates that anaerobic bacteria and known halophilic taxa are present and viable in the fracture waters presently outflowing from existing boreholes. Major cations and anions found in the fracture waters at the 2.4 km level of the mine are also reported.


Continued surveillance of Serratia marcescens infections by bacteriocin typing: investigation of two outbreaks of cross-infection in an intensive care unit.
During an 8.5-month period, 198 additional isolates of Serratia marcescens were typed by bacteriocin sensitivity; 154 isolates were typable and were categorized according to our current system of 54 provisional bacteriocin sensitivity patterns. Two outbreaks of nosocomial infection due to S. marcescens occurred in our intensive care unit, involving two and five patients, respectively. The latter outbreak was caused by a strain of S. marcescens which was not sensitive to any of the 10 bacteriocins normally used. Therefore we developed a supplementary procedure based on bacteriocin production rather than bacteriocin sensitivity. Bacteriocin production was induced with mitomycin C, and the crude lysates were applied to 15 provisional bacteriocin indicator strains. The reverse typing procedure was necessary to determine the spread and ultimate subsidence of this particular outbreak of cross-infection.


Addressing Unmet Medical Needs in Type 1 Diabetes: A Review of Drugs Under Development.
The incidence of type 1 diabetes (T1D) is increasing worldwide and there is a very large need for effective therapies. Essentially no therapies other than insulin are currently approved for the treatment of T1D. Drugs already in use for type 2 diabetes and many new drugs are under clinical development for T1D, including compounds with both established and new mechanisms of action. Content of the Review: Most of the new compounds in clinical development are currently in Phase 1 and 2. Drug classes discussed in this review include new insulins, SGLT inhibitors, GLP-1 agonists, immunomodulatory drugs including autoantigens and anti-cytokines, agents that regenerate β-cells and others. Regulatory Considerations: In addition, considerations are provided with regard to the regulatory environment for the clinical development of drugs for T1D, with a focus on the United States Food and Drug Administration and the European Medicines Agency. Future opportunities, such as combination treatments of immunomodulatory and beta-cell regenerating therapies, are also discussed.


End of induction positron emission tomography complete response (PET-CR) as a surrogate for progression-free survival in previously untreated follicular lymphoma.
Progression-free survival (PFS) has been the regulatory primary end-point for recent phase III trials in first-line follicular lymphoma (FL), but requires prolonged follow-up. Complete response (CR) at 30 months after initiation of induction treatment was validated as surrogate end-point for PFS. Our objective was to further evaluate surrogacy of CR measured by [18 F] fluorodeoxyglucose (FDG) positron emission tomography (PET) imaging at the end of induction (EoI). Individual patient data were analysed from 1505 patients from five randomized trials. Trial-level surrogacy examining the association between treatment effects on EoI-PET-CR and PFS was evaluated using linear regression ( RWLS2 ) and bivariate Copula ( RCopula2 ) models. Although EoI-PET-CR strongly predicted PFS at a prognostic level, the trial-level assessment did not show strong correlation ( RWLS2=0.56 , confidence interval [CI]: 0.20-0.88; RCopula2=0.35 , CI: 0.0-0.82). The high uncertainty in estimation was possibly due to the small number of trials and the population of patients with available PET data. Maintenance therapy affecting PFS beyond induction treatment, but not EoI-PET-CR end-point, may have distorted the association between treatment effects. However, there will probably be a number of additional trials approaching completion with available PET response data. Refined evaluation of PET-CR based surrogate end-points is still warranted.


Serratus anterior and lower trapezius muscle activities during multi-joint isotonic scapular exercises and isometric contractions.
Proper scapular function during humeral elevation, such as upward rotation, external rotation, and posterior tilting of the scapula, is necessary to prevent shoulder injury. However, the appropriate intensity of rehabilitation exercise for the periscapular muscles has yet to be clarified. To identify the serratus anterior, lower trapezius, infraspinatus, and posterior deltoid muscle activities during 2 free-motion exercises using 3 intensities and to compare these muscle activities with isometric contractions during quadruped shoulder flexion and external rotation and abduction of the glenohumeral joint. Cross-sectional study. Health Science Laboratory. A total of 16 uninjured, healthy, active, male college students (age = 19.5 ± 1.2 years, height = 173.1 ± 6.5 cm, weight = 68.8 ± 6.6 kg). Mean electromyographic activity normalized by the maximal voluntary isometric contraction was analyzed across 3 intensities and 5 exercises. Intraclass correlation coefficients were calculated for electromyographic activity of the 4 muscles in each free-motion exercise. Significant interactions in electromyographic activity were observed between intensities and exercises (P < .05). The quadruped shoulder-flexion exercise activated all 4 muscles compared with other exercises. Also, the modified robbery free-motion exercise activated the serratus anterior, lower trapezius, and infraspinatus compared with the lawn-mower free-motion exercise. However, neither exercise showed a difference in posterior deltoid electromyographic activity. Three intensities exposed the nature of the periscapular muscle activities across the different exercises. The free-motion exercise in periscapular muscle rehabilitation may not modify serratus anterior, lower trapezius, and infraspinatus muscle activities unless knee-joint extension is limited.


A randomized, placebo-controlled trial of lubiprostone for opioid-induced constipation in chronic noncancer pain.
This multicenter, phase 3 trial evaluated oral lubiprostone for constipation associated with non-methadone opioids in patients with chronic noncancer-related pain. Adults with opioid-induced constipation (OIC; <3 spontaneous bowel movements [SBMs] per week) were randomized 1:1 to double-blind lubiprostone 24 μg or placebo twice daily for 12 weeks. The primary end point was the overall SBM response rate. Responders had at least moderate response (≥1 SBM improvement over baseline frequency) in all treatment weeks with available observed data, as well as full response (≥3 SBMs per week) for at least 9 of the 12 treatment weeks. In total, 431 patients were randomized; 212 each received lubiprostone and placebo, and 7 were not treated. Overall, the SBM response rate was significantly higher for patients treated with lubiprostone vs. placebo (27.1 vs. 18.9%, respectively; P=0.030). Overall mean change from baseline in SBM frequency was significantly greater with lubiprostone vs. placebo (3.2 vs. 2.4, respectively; P=0.001). The median time to first SBM was significantly shorter with lubiprostone vs. placebo (23.5 vs. 37.7 h, respectively; P=0.004). Compared with placebo, the patients treated with lubiprostone exhibited significant improvements in straining (P=0.004), stool consistency (P<0.001), and constipation severity (P=0.010). No significant differences were observed in quality-of-life measures or the use of rescue medication; however, the percentage of patients who used rescue medication was consistently lower in the lubiprostone group than in the placebo group at months 1 (34.9 vs. 37.7%), 2 (23.4 vs. 26.6%), and 3 (20.5 vs. 22.0%). Adverse events (AEs) >5% were diarrhea, nausea, vomiting, and abdominal pain (lubiprostone: 11.3, 9.9, 4.2, and 7.1%, respectively; placebo, 3.8, 4.7, 5.2, and 0%, respectively). None of the serious AEs (lubiprostone, 3.3%; placebo, 2.8%) were related to lubiprostone. Lubiprostone significantly improved symptoms of OIC and was well tolerated in patients with chronic noncancer pain.


Novel agents in the treatment of pancreatic adenocarcinoma.
The development of new agents and treatment strategies against pancreatic adenocarcinoma is vital, given the poor prognosis of the patients with this particular type of cancer. Three novel compounds were tested at different phases of clinical research and the results were presented in the recent ASCO Gastrointestinal Cancers Symposium. FG-3019 inhibits the connective tissue growth factor which is important in the biology of pancreatic cancer and was shown to be relatively safe in a phase I study (Abstract #213). In addition, ASG-5ME, an antibody specific for SLC44A4 that is universally expressed in pancreatic cancer and also carries a conjugate chemotherapy particle was safe at the appropriate dosing in a phase I trial (Abstract #176). Last but not least, tanespimycin, a molecule that inhibits heat shock protein 90 was not effective in the first line treatment of patients with pancreatic cancer in a phase II study (Abstract #245). Further studying of FG-3019 and ASG-5ME will show the potential activity if any of these compounds in patients with pancreatic cancer.


DSM-IV criteria for childhood separation anxiety disorder: informant, age, and sex differences.
The present study examines frequency of DSM-IV symptom and diagnostic criteria for separation anxiety disorder (SAD) by informant, age, and sex. Children aged 4-15 years with a primary DSM-IV diagnosis of SAD (N=106) were assessed using structured diagnostic interviews (Kinder-DIPS; DSM-IV-TR Version). Frequency of DSM-IV symptom and diagnostic criteria were examined as a function of informant and child characteristics, along with impairment and distress ratings. The most frequently reported symptoms were separation-related distress, avoidance of being alone/without an adult and sleeping away from caregivers or from home, with nightmares the least frequently endorsed criterion. Child report did not yield any significant sex or age differences. However, parent report revealed greater reluctance or avoidance of school attendance for girls than boys, and for younger children (<8 years). Parent report indicated greater symptom-related impairment than child report, and the number of symptoms was correlated with impairment based on parent report, and with distress based on child report. The primary indicators of SAD appear to be separation distress, avoidance of being alone, and sleeping away from caregivers. Findings suggest that parents may be best placed to determine impairment, while children may be the most accurate reporters of more covert internal distress. Implications for clinicians are that reports from multiple informants should be used to gain the most comprehensive information about childhood SAD.


Modulation of Renal GLUT2 by the Cannabinoid-1 Receptor: Implications for the Treatment of Diabetic Nephropathy.
Altered glucose reabsorption via the facilitative glucose transporter 2 (GLUT2) during diabetes may lead to renal proximal tubule cell (RPTC) injury, inflammation, and interstitial fibrosis. These pathologies are also triggered by activating the cannabinoid-1 receptor (CB1R), which contributes to the development of diabetic nephropathy (DN). However, the link between CB1R and GLUT2 remains to be determined. Here, we show that chronic peripheral CB1R blockade or genetically inactivating CB1Rs in the RPTCs ameliorated diabetes-induced renal structural and functional changes, kidney inflammation, and tubulointerstitial fibrosis in mice. Inhibition of CB1R also downregulated GLUT2 expression, affected the dynamic translocation of GLUT2 to the brush border membrane of RPTCs, and reduced glucose reabsorption. Thus, targeting peripheral CB1R or inhibiting GLUT2 dynamics in RPTCs has the potential to treat and ameliorate DN. These findings may support the rationale for the clinical testing of peripherally restricted CB1R antagonists or the development of novel renal-specific GLUT2 inhibitors against DN.


Seventeenth-century telescope optics of Torricelli, Divini, and Campani.
Optics of telescope makers Torricelli, Divini, and Campani dating back to the 17th century have been tested interferometrically. It was found that the optical polishing technology had progressed to thorough control over the surface figure and finish, while the nearly paraxial conditions of the aperture remained constant. The instruments that were examined exemplify the knowledge of optics of that time, particularly with respect to the early use of erector units in terrestrial telescopes.


Surgery in patients with acquired immunodeficiency syndrome.
Between 1982 and 1985, 21 patients with acquired immunodeficiency syndrome (20 men and one woman; mean age, 36 years) underwent 31 surgical procedures at the Harbor/UCLA Medical Center, Torrance, or the UCLA Medical Center (skin, lymph node, and endoscopic biopsies were excluded). The operations included seven emergencies and 24 elective operations (eight major and 16 minor). Pathologic findings included cytomegalovirus colon perforation (two), disseminated Kaposi's sarcoma (KS) of the small and large bowel (one), cystic duct obstruction by KS (one), poorly differentiated gastrointestinal lymphoma (one), Candida acalculous cholecystitis (one), central nervous system toxoplasmosis (two), amebic encephalitis with abscess (one), staphylococcal botryomycosis of the pericardium (one), pulmonary KS (one), and cytomegalovirus (one). The overall operative (30 days) mortality rate was 48% (10/21). The emergency surgery rate was 57% (4/7), elective, 43% (6/14). The high operative mortality rate in these patients was usually due to progression of opportunistic infections or malignancy.


Advantages and techniques of utilizing anterolateral portal in delaminated rotator cuff repair.
Typically, rotator cuff repair is performed two-dimensionally while visualizing the subacromial space. To achieve a more complete repair, sutures can be retrieved from inside the joint utilizing a penetrating suture device through the anterolateral portal, which permits visualization of the articular side and bursal side of the rotator cuff tear. Utilizing other portals can leave the sutures out of sight and reach. The anterolateral portal helps capture both leaves of a delaminated tear and places the sutures in the center of the field for retrieval and tying. The anterolateral portal is located with a spinal needle just off the anterolateral corner of the acromion, and the suture anchor is inserted into the greater tuberosity through this portal. The sutures are passed into the joint through the tear in the rotator cuff with a suture grasper. The scope is redirected intraarticularly, and the suture is visualized. The sutures are positioned, and the penetrator is passed through the tendon. The scope is placed back in the bursa for tying sutures and capturing both leaves of a delaminated tear. It works equally well for simple complete tears and partial articular-sided supraspinatus tendon avulsion lesions.


Structure-function analysis of NADPH:nitrate reductase from Aspergillus nidulans: analysis of altered pyridine nucleotide specificity in vivo.
Nitrate reductase (NaR) catalyses the reduction of nitrate to nitrite via a two-electron transfer. In fungi, the electron donor for NaR is NADPH whereas plants can have two enzymes, NADH:NaR and a bispecific NAD(P)H:NaR. PCR mutagenesis was employed to introduce mutations into the niaD gene of Aspergillus nidulans in order to identify residues involved in co-enzyme specificity. The niaD3000 mutation (NiaD T813D, K814Q) altered co-enzyme specificity: the new enzyme had high levels of NADH:NaR activity in vitro, whilst all NADPH-associated activity was lost. However, strains carrying this mutation did not grow on nitrate. Enzyme assays suggested that this was not due to inhibition of the mutant enzyme by NADPH. All revertants of the niaD3000 mutants had restored NADPH activity and lost NADH activity. Sequence analysis of these revertants showed that they all contained a single amino acid change at Asp-813, suggesting that this position is crucial to co-enzyme specificity. Further studies have shown that the mutant enzyme was not protected from deactivation by either co-factor in cell-free extracts (unlike the wild-type), and that induction of the glucose-6-phosphate dehydrogenase occurred independently of NADPH levels. These data highlight the importance of functional tests in vivo under physiological conditions.


Association between body mass index and urinary tract infection in adult patients.
Overweight and obesity have become a global public health over the last decades. Obesity has been suggested to be a risk factor for some infections, but studies often showed controversial findings. Few studies examined the relationship between body mass index (BMI) and urinary tract infection (UTI), showing inconsistent results. The purpose of this study was to determine the relationship between BMI and UTI in adult patients. Adult patients (≥ 18 years old) who were referred to clinics or admitted in hospital with diagnosis of UTI were considered for participation in the study. Control group were selected from healthy adult normal population whom underwent medical check-ups at the same hospital and without history of UTI. Data about age, gender, history of diabetes mellitus and BMI were registered for individuals who met inclusion criteria. A total of 116 patients with UTI and 156 people as the control group were included in the study. Two groups were matched for age, gender and history of diabetes mellitus. Mean BMI ± SD of patients was 25.2 ± 4.0 kg/m(2) and the controls was 25.1 ± 3.6 kg/m(2). There was no significant correlation between BMI and UTI (P = 0.757). Mean BMI ± SD of patients with upper UTI was 25.6 ± 4.1 kg/m(2) and for lower UTI was 24.9 ± 4.0 kg/m(2). There was no significant difference between BMI of controls and patients with any type of UTI (P = 0.573). Our findings did not found an association between BMI and UTI and does not support obesity as a risk factor for UTI in adult patients.


Metastatic Intracerebral Plasmacytoma Treated with Radiation and Thalidomide, Dexamethasone with Cyclophosphamide Chemotherapy.
Intracerebral plasmacytoma is an extremely rare disease for which no treatment protocol has been established. The authors present a case of metastatic intracerebral plasmacytoma that was partially resected and treated with radiation therapy. For tumor recurrence, a combination chemotherapy regimen was initiated. A 48-year-old male patient presented with dizziness and memory loss. Conventional magnetic resonance imaging (MRI) of the brain revealed a multilobulated mass with enhancing cystic and solid components measuring 7 × 7 × 6 cm in the left frontal lobe. The patient had undergone subtotal gastrectomy and transverse colectomy 8 years before admission and had been diagnosed with extraosseous plasmacytoma. At the time of the current presentation, the patient underwent craniotomy for the parenchymal lesion. Partial tumor resection was performed. Histologic and immunohistochemical examinations confirmed the diagnosis of plasmacytoma. Fractionated radiotherapy was administered, and no enhancing mass was observed on follow-up MRI. One year after radiotherapy, tumor recurrence was observed in a different area of the cerebral parenchyma. Combination thalidomide, dexamethasone, and cyclophosphamide chemotherapy was administered. After three cycles of chemotherapy, the tumor was well controlled on MRI. Hereafter, two more times of tumor recurrence occurred in the other sites of the cerebral parenchyma, but with chemoradiation therapy, the tumor was well suppressed. The findings of this case suggest that the cerebral parenchyma can be one of the metastatic sites for extraosseous plasmacytoma. In addition, combination chemotherapy with thalidomide, dexamethasone, and cyclophosphamide may be a useful treatment option for intracerebral plasmacytoma.


Ethnic disparities in the risk of colorectal adenomas associated with aspirin and statin use: a retrospective multiethnic study.
Although data on the inverse association between colorectal adenomas (CRA) and daily aspirin or statin therapy exists in white and black patients, scarce data exists on these associations in the Hispanic population. With a rapidly increasing Hispanic population in the United States, defining the association in Hispanics is crucial. The study sample included 1,843 consecutive patients who underwent a colonoscopy (screening or diagnostic) from 2009 to 2011 at a community hospital in East Meadow, New York. Data was then extracted from patient charts regarding aspirin and/or statin use. Adjusted odds ratios (OR) and their 95% confidence intervals (CI) were calculated to assess the association between colonoscopy findings and aspirin, statin, or aspirin/statin use. In our total population including all races, aspirin user had an increased risk for having two or more adenomas (OR =1.73, 95% CI: 1.00, 2.99, P=0.05) and presence of an adenoma in the proximal colon (OR =1.66, 95% CI: 1.07, 2.58, P=0.02). In the total study population, those who used both statin and aspirin had an increased risk for having two or more adenomas (OR =2.56, 95% CI: 1.21, 5.39, P=0.01). In the Hispanic population, users of both medications had an increased risk for having two or more adenomas (OR =19.04, 95% CI: 1.30, 280.09, P=0.03), adenoma present in the distal colon (OR =5.75, 95% CI: 1.64, 20.21, P=0.01) and largest adenoma in distal colon (OR =5.75, 95% CI: 1.64, 20.21, P=0.01). Aspirin use and aspirin/statin use was associated with abnormal colonoscopy findings, particularly in the Hispanic population. These findings may be due to environmental factors such as dietary, colonic flora, or genetic susceptibility. The findings warrant further investigational research, particularly in Hispanics.


[Capsaicin-sensitive afferents in the vagus nerve].
Vanilloids were shown to interact with over 70% of vagal C-afferents first causing an excitation followed by desensitisation and a lasting destruction of nerve fibres. Capsaicin induces a secretion of some neuropeptides from 10-30% of vagal sensory terminals and therefore serves as a pharmacological tool for testing local "effector function" of primary afferents. Vagal afferents seem to have their own subtype of vanilloid receptors (VR), not completely identical with the VR receptors in the dorsal root ganglia. Considering potentiation of the capsaicin receptors sensitivity by some factors such as local heating, pH, free oxygen radicals, a possible role of the VRs as integrators of chemical and physical components of nociceptive stimuli, is discussed.


Accuracy of genomic prediction of purebreds for cross bred performance in pigs.
In pig breeding, as the final product is a cross bred (CB) animal, the goal is to increase the CB performance. This goal requires different strategies for the implementation of genomic selection from what is currently implemented in, for example dairy cattle breeding. A good strategy is to estimate marker effects on the basis of CB performance and subsequently use them to select pure bred (PB) breeding animals. The objective of our study was to assess empirically the predictive ability (accuracy) of direct genomic values of PB for CB performance across two traits using CB and PB genomic and phenotypic data. We studied three scenarios in which genetic merit was predicted within each population, and four scenarios where PB genetic merit for CB performance was predicted based on either CB or a PB training data. Accuracy of prediction of PB genetic merit for CB performance based on CB training data ranged from 0.23 to 0.27 for gestation length (GLE), whereas it ranged from 0.11 to 0.22 for total number of piglets born (TNB). When based on PB training data, it ranged from 0.35 to 0.55 for GLE and from 0.30 to 0.40 for TNB. Our results showed that it is possible to predict PB genetic merit for CB performance using CB training data, but predictive ability was lower than training using PB training data. This result is mainly due to the structure of our data, which had small-to-moderate size of the CB training data set, low relationship between the CB training and the PB validation populations, and a high genetic correlation (0.94 for GLE and 0.90 for TNB) between the studied traits in PB and CB individuals, thus favouring selection on the basis of PB data.


Insulin secretion enhancing activity of roselle calyx extract in normal and streptozotocin-induced diabetic rats.
Our recent study revealed the antihyperglycemic activity of an ethanolic extract of roselle calyxes (Hibiscus sabdariffa) in diabetic rats. The present study had, therefore, an objective to investigate the mechanism underlying this activity. Male Sprague Dawley rats were induced to be diabetes by intraperitoneal injection of 45 mg/kg streptozotocin (STZ). Normal rats as well as diabetic rats were administered with the ethanolic extract of H. sabdariffa calyxes (HS-EE) at 0.1 and 1.0 g/kg/day, respectively, for 6 weeks. Then, blood glucose and insulin levels, at basal and glucose-stimulated secretions, were measured. The pancreas was dissected to examine histologically. HS-EE 1.0 g/kg/day significantly decreased the blood glucose level by 38 ± 12% in diabetic rats but not in normal rats. In normal rats, treatment with 1.0 g/kg HS-EE increased the basal insulin level significantly as compared with control normal rats (1.28 ± 0.25 and 0.55 ± 0.05 ng/ml, respectively). Interestingly, diabetic rats treated with 1.0 g/kg HS-EE also showed a significant increase in basal insulin level as compared with the control diabetic rats (0.30 ± 0.05 and 0.15 ± 0.01 ng/ml, respectively). Concerning microscopic histological examination, HS-EE 1.0 g/kg significantly increased the number of islets of Langerhans in both normal rats (1.2 ± 0.1 and 2.0 ± 0.1 islet number/10 low-power fields (LPF) for control and HS-EE treated group, respectively) and diabetic rats (1.0 ± 0.3 and 3.9 ± 0.6 islet number/10 LPF for control and HS-EE treated group, respectively). The antidiabetic activity of HS-EE may be partially mediated via the stimulating effect on insulin secretion.


"It started 30 years ago, and it still haunts me": an exploratory investigation of Territorians' gambling behaviours, harm, and help-seeking for gambling issues in an Australian jurisdiction.
There is a lack of qualitative gambling research on lived experience, help-seeking, and gamblers and affected others' views on the regulatory environment in the Northern Territory (NT), Australia. This study provides 1) lived experience of individuals who reported experiencing harms from gambling, 2) insights into help-seeking for gambling issues, 3) and people's views on current legislation on gambling in the NT. The results of this study begin to establish an evidence base that could be used to inform targeted interventions for people experiencing harms from gambling in the NT. Semi-structured interviews were conducted with a targeted selection of respondents from the 2015 and 2018 NT Gambling Prevalence and Wellbeing Surveys. The sample (n = 27; age 18+ years; Aboriginal (The term of 'Aboriginal' has used been used throughout the manuscript to reflect Aboriginal and Torres Strait Islander, Indigenous, or First Nations people for purposes of brevity. We respect the diversity among these populations.) and Non-Aboriginal) included weekly (online and venue-based electronic gambling machine (EGM)) gamblers, non-regular gamblers, and those negatively affected by others' gambling. A Framework Analysis approach was used for data analysis. Appropriate ethics approval was obtained. Negative impacts and harms from gambling were experienced by both gamblers and non-gamblers. These included monetary losses, relationship conflicts, emotional distress, and decrements to health. A lack of self-realisation of gambling issues and awareness of the available services, shame, and embarrassment, were reported as the main barriers to help-seeking. Where help was sought, it was primarily informal (e.g., family) and was rarely preventive. In many instances, self-help strategies were successful in controlling one's own gambling. Gamblers suggested regulations should set limits on the daily number of hours of playing, the bet size, and reduced access to EGM. The need for strengthening the existing awareness and education interventions was emphasised. Viewing the findings from a public health lens, targeted approaches based on specific circumstances may have the potential to minimise harms from gambling, but only for those already experiencing harms. The treatment, policy, and regulatory approaches need to be tailored to address the causes and impacts of harms experienced by people.


Mirror Therapy in Stroke Rehabilitation: Current Perspectives.
In contrast to varied therapy approaches, mirror therapy (MT) can be used even in completely plegic stroke survivors, as it uses visual stimuli for producing a desired response in the affected limb. MT has been studied to have effects not just on motor impairments but also on sensations, visuospatial neglect, and pain after stroke. This paper attempts to systematically review and present the current perspectives on mirror therapy and its application in stroke rehabilitation, and dosage, feasibility and acceptability in stroke rehabilitation. An electronic database search across Google, PubMed, Web of Science, etc., generated 3871 results. After screening them based on the inclusion and exclusion criteria, we included 28 studies in this review. The data collected were divided on the basis of application in stroke rehabilitation, modes of intervention delivery, and types of control and outcome assessment. We found that most studies intervened for upper limb motor impairments post stroke. Studies were equally distributed between intervention in chronic and acute phases post stroke with therapy durations lasting between 1 and 8 weeks. MT showed definitive motor and sensory improvements although the extent of improvements in sensory impairments and hemineglect is limited. MT proves to be an effective and feasible approach to rehabilitate post-stroke survivors in the acute, sub-acute, and chronic phases of stroke, although its long-term effects and impact on activities of daily living need to be analysed extensively.


Weighted Gene Co-expression Network Analysis Identifies Critical Genes for the Production of Cellulase and Xylanase in Penicillium oxalicum.
Genes involved in cellular processes undergo environment-dependent co-regulation, but the co-expression patterns of fungal cellulase and xylanase-encoding genes remain unclear. Here, we identified two novel carbon sources, methylcellulose and 2-hydroxyethyl cellulose, which efficiently induced the secretion of cellulases and xylanases in Penicillium oxalicum. Comparative transcriptomic analyses identified carbon source-specific transcriptional patterns, mainly including major cellulase and xylanase-encoding genes, genes involved in glycolysis/gluconeogenesis and the tricarboxylic acid cycle, and genes encoding transcription factors, transporters and G protein-coupled receptors. Moreover, the weighted correlation network analysis of time-course transcriptomes, generated 17 highly connected modules. Module MEivory, comprising 120 members, included major cellulase and xylanase-encoding genes, genes encoding the key regulators PoxClrB and PoxXlnR, and a cellodextrin transporter POX06051/CdtC, which were tightly correlated with the filter-paper cellulase, carboxymethylcellulase and xylanase activities in P. oxalicum. An expression kinetic analysis indicated that members in MEivory were activated integrally by carbon sources, but their expressional levels were carbon source- and/or induction duration-dependent. Three uncharacterized regulatory genes in MEivory were identified, which regulate the production of cellulases and xylanases in P. oxalicum. These findings provide insights into the mechanisms associated with the synthesis and secretion of fungal cellulases and xylanases, and a guide for P. oxalicum application in biotechnology.


Second-line treatment persistence and costs among patients with immune-mediated rheumatic diseases treated with subcutaneous TNF-alpha inhibitors.
The objective of this study was to describe treatment persistence with second-line subcutaneous tumor necrosis factor-alpha inhibitors (SC-TNFis) in patients with immune-mediated rheumatic diseases (IMRDs) in Sweden, and the impact of non-persistence on healthcare costs. This retrospective observational study was based on Swedish national health register data. Adults were identified through filled prescriptions for adalimumab (ADA), etanercept (ETA), certolizumab pegol (CZP) and golimumab (GLM). Persistence was estimated over 3 years for propensity score-matched (PSM) cohorts using non-parametric survival analysis. Unadjusted comparisons of costs comprised specialized outpatient care, inpatient care, and medication. In total, N = 845 patients were identified and three PSM cohorts were generated (GLM vs. ADA, ETA, and CZP, respectively). GLM exhibited higher persistence than ADA over the study period (p = 0.040), and numerically higher persistence than ETA and CZP for 36 and 30 months, respectively. Persistent and non-persistent patients had similar mean total cost at 12 month pre-treatment ($5185 vs. $5064, p = 0.750). During the 12 month post-treatment initiation, persistent patients had lower mean total costs ($4377 vs. $6605), corresponding to a cost difference of $2228 (p < 0.001). In second-line treatment with SC-TNFis for IMRDs in Sweden, GLM exhibited significantly higher persistence than ADA over the course of the study. Similarly, GLM showed numerically higher persistence than ETA and CZP, which is concurrent with results observed in first-line SC-TNFi treatment. Considering the lower healthcare costs for persistent patients, the choice of second-line SC-TNFi among eligible patients may merit careful consideration given its impact on patients and payers.


Journal article content as a predictor of commitment to change among continuing medical education respondents.
Journal reading is a time-tested means of continuing medical education (CME) among physicians. Asking physicians to make a written statement of commitment to change has been shown to increase the likelihood of eventual change in practice behavior when used in conjunction with CME lectures. We describe the type and medical content of articles responsible for commitment to change comments among readers of a large circulation primary care journal. Response to the question "what change(s) do you plan to make in your practice as a result of reading the articles in this issue?" were analyzed from CME response cards associated with six issues (1 year's publication) of the Archives of Family Medicine. Responses indicating a commitment to change were analyzed to characterize the type and content of articles responsible for their generation. During the 1-year study period, original contributions (reports of research trials) dealing with medicine and preventive medicine content accounted for the greatest number of comments. After adjusting for the frequency of article type and content, special articles and those dealing with complementary medicine accounted for the highest number of comments on a per-article basis. Family physicians make commitment to change statements on the basis of a broad range of journal articles. Certain articles are more likely to generate statements of commitment to planned changes in practice.


Cytosolic Innate Immune Sensing and Signaling upon Infection.
Cytosolic sensing of pathogens is essential to a productive immune response. Recent reports have emphasized the importance of signaling platforms emanating from organelles and cytosolic sensors, particularly during the response to intracellular pathogens. Here, we highlight recent discoveries identifying the key mediators of nucleic acid and cyclic nucleotide sensing and discuss their importance in host defense. This review will also cover strategies evolved by pathogens to manipulate these pathways.


Reduced lysosomal acid lipase activity: A new marker of liver disease severity across the clinical continuum of non-alcoholic fatty liver disease?
Lysosomal acid lipase (LAL) plays a key role in intracellular lipid metabolism. Reduced LAL activity promotes increased multi-organ lysosomal cholesterol ester storage, as observed in two recessive autosomal genetic diseases, Wolman disease and Cholesterol ester storage disease. Severe liver steatosis and accelerated liver fibrosis are common features in patients with genetic LAL deficiency. By contrast, few reliable data are available on the modulation of LAL activity in vivo and on the epigenetic and metabolic factors capable of regulating its activity in subjects without homozygous mutations of the Lipase A gene. In the last few years, a less severe and non-genetic reduction of LAL activity was reported in children and adults with non-alcoholic fatty liver disease (NAFLD), suggesting a possible role of LAL reduction in the pathogenesis and progression of the disease. Patients with NAFLD show a significant, progressive reduction of LAL activity from simple steatosis to non-alcoholic steatohepatitis and cryptogenic cirrhosis. Among cirrhosis of different etiologies, those with cryptogenic cirrhosis show the most significant reductions of LAL activity. These findings suggest that the modulation of LAL activity may become a possible new therapeutic target for patients with more advanced forms of NAFLD. Moreover, the measurement of LAL activity may represent a possible new marker of disease severity in this clinical setting.


Characterization of Fruit Development, Antioxidant Capacity, and Potential Vasoprotective Action of Peumo (Cryptocarya alba), a Native Fruit of Chile.
The peumo (Cryptocarya alba) is a native fruit from central Chile that belongs to the Lauraceae family. To characterize the development and the potential health benefits of this edible fruit, quality and physiological parameters, along with antioxidant capacity, were evaluated during three clearly defined developmental stages of the fruit in two seasons. The most distinguishable attributes of ripe fruit were the change in size and color. Low CO2 production and no detectable ethylene levels suggested non-climacteric behavior of the peumo fruit. Peumo demonstrate a significant increase in their antioxidant capacity per 1 g of fresh weight (FW) of the sample, from small to ripe fruit. Higher values in ripe fruit (FRAP: 37.1-38.3 µmol FeSO4/gFW, TEAC: 7.9-8.1 mmol TE/gFW, DPPH: 8.4-8.7 IC50 μg/mL, and ORAC: = 0.19-0.20 mmol TE/gFW) were observed than those in blueberry fruit (FRAP: 4.95 µmol FeSO4/gFW, TEAC: 1.25 mmol TE/gFW, DPPH: 11.3 IC50 μg/mL, and ORAC: 0.032 mmol TE/ gFW). The methanol extracts of ripe fruit displayed the presence of polyphenol acids and quercetin, an ORAC value of 0.637 ± 0.061 mmol TE per g dried weight (DW), and a high cellular antioxidant and anti-inflammatory potential, the latter exceeding the effect of quercetin and indomethacin used as standard molecules. Also, the assay of isolated rat aorta with endothelium-dependent relaxation damage demonstrated that the peumo extract induced vascular protection, depending on its concentration under a high glucose condition. These results demonstrate that these endemic fruits have a good chance as ingredients or foods with functional properties.


Evaluation of the role of STAP1 in Familial Hypercholesterolemia.
Familial hypercholesterolemia (FH) is characterised by elevated serum levels of low-density lipoprotein cholesterol (LDL-C) and a substantial risk for cardiovascular disease. The autosomal-dominant FH is mostly caused by mutations in LDLR (low density lipoprotein receptor), APOB (apolipoprotein B), and PCSK9 (proprotein convertase subtilisin/kexin). Recently, STAP1 has been suggested as a fourth causative gene. We analyzed STAP1 in 75 hypercholesterolemic patients from Berlin, Germany, who are negative for mutations in canonical FH genes. In 10 patients with negative family history, we additionally screened for disease causing variants in LDLRAP1 (low density lipoprotein receptor adaptor protein 1), associated with autosomal-recessive hypercholesterolemia. We identified one STAP1 variant predicted to be disease causing. To evaluate association of serum lipid levels and STAP1 carrier status, we analyzed 20 individuals from a population based cohort, the Cooperative Health Research in South Tyrol (CHRIS) study, carrying rare STAP1 variants. Out of the same cohort we randomly selected 100 non-carriers as control. In the Berlin FH cohort STAP1 variants were rare. In the CHRIS cohort, we obtained no statistically significant differences between carriers and non-carriers of STAP1 variants with respect to lipid traits. Until such an association has been verified in more individuals with genetic variants in STAP1, we cannot estimate whether STAP1 generally is a causative gene for FH.


Bucillamine prevents cisplatin-induced ototoxicity through induction of glutathione and antioxidant genes.
Bucillamine is used for the treatment of rheumatoid arthritis. This study investigated the protective effects of bucillamine against cisplatin-induced damage in auditory cells, the organ of Corti from postnatal rats (P2) and adult Balb/C mice. Cisplatin increases the catalytic activity of caspase-3 and caspase-8 proteases and the production of free radicals, which were significantly suppressed by pretreatment with bucillamine. Bucillamine induces the intranuclear translocation of Nrf2 and thereby increases the expression of γ-glutamylcysteine synthetase (γ-GCS) and glutathione synthetase (GSS), which further induces intracellular antioxidant glutathione (GSH), heme oxygenase 1 (HO-1) and superoxide dismutase 2 (SOD2). However, knockdown studies of HO-1 and SOD2 suggest that the protective effect of bucillamine against cisplatin is independent of the enzymatic activity of HO-1 and SOD. Furthermore, pretreatment with bucillamine protects sensory hair cells on organ of Corti explants from cisplatin-induced cytotoxicity concomitantly with inhibition of caspase-3 activation. The auditory-brainstem-evoked response of cisplatin-injected mice shows marked increases in hearing threshold shifts, which was markedly suppressed by pretreatment with bucillamine in vivo. Taken together, bucillamine protects sensory hair cells from cisplatin through a scavenging effect on itself, as well as the induction of intracellular GSH.


Integrated proteomics identified up-regulated focal adhesion-mediated proteins in human squamous cell carcinoma in an orthotopic murine model.
Understanding the molecular mechanisms of oral carcinogenesis will yield important advances in diagnostics, prognostics, effective treatment, and outcome of oral cancer. Hence, in this study we have investigated the proteomic and peptidomic profiles by combining an orthotopic murine model of oral squamous cell carcinoma (OSCC), mass spectrometry-based proteomics and biological network analysis. Our results indicated the up-regulation of proteins involved in actin cytoskeleton organization and cell-cell junction assembly events and their expression was validated in human OSCC tissues. In addition, the functional relevance of talin-1 in OSCC adhesion, migration and invasion was demonstrated. Taken together, this study identified specific processes deregulated in oral cancer and provided novel refined OSCC-targeting molecules.


Fine-scale genetic characterization of Plasmodium falciparum chromosome 7 encompassing the antigenic var and the drug-resistant pfcrt genes.
The fact that malaria is still an uncontrolled disease is reflected by the genetic organization of the parasite genome. Efforts to curb malaria should begin with proper understanding of the mechanism by which the parasites evade human immune system and evolve resistance to different antimalarial drugs. We have initiated such a study and presented herewith the results from the in silico understanding of a seventh chromosomal region of the malarial parasite Plasmodium falciparum encompassing the antigenic var genes (coding pfemp1) and the drug-resistant gene pfcrt located at a specified region of the chromosome 7. We found 60 genes of various functions and lengths, majority (61.67%) of them were performing known functions. Almost all the genes have orthologs in other four species of Plasmodium, of which P. chabaudi seems to be the closest to P. falciparum. However, only two genes were found to be paralogous. Interestingly, the drug-resistant gene, pfcrt was found to be surrounded by seven genes coding for several CG proteins out of which six were reported to be responsible for providing drug resistance to P. vivax. The intergenic regions, in this specified region were generally large in size, majority (73%) of them were of more than 500 nucleotide bp length. We also designed primers for amplification of 21 noncoding DNA fragments in the whole region for estimating genetic diversity and inferring the evolutionary history of this region of P. falciparum genome.


The mercurial nature of neutrophils: still an enigma in ARDS?
The acute respiratory distress syndrome (ARDS) is a life-threatening lung condition resulting from direct and indirect insults to the lung. It is characterized by disruption of the endothelial-epithelial barrier, alveolar damage, pulmonary edema, and respiratory failure. A key feature of ARDS is the accumulation of neutrophils in the lung microvasculature, interstitium, and alveolar space. Despite a clear association between neutrophil influx into the lung and disease severity, there is some debate as to whether neutrophils directly contribute to disease pathogenesis. The primary function of neutrophils is to provide immediate host defense against pathogenic microorganisms. Neutrophils release numerous antimicrobial factors such as reactive oxygen species, proteinases, and neutrophil extracellular traps. However, these factors are also toxic to host cells and can result in bystander tissue damage. The excessive accumulation of neutrophils in ARDS may therefore contribute to disease progression. Central to neutrophil recruitment is the release of chemokines, including the archetypal neutrophil chemoattractant IL-8, from resident pulmonary cells. However, the chemokine network in the inflamed lung is complex and may involve several other chemokines, including CXCL10, CCL2, and CCL7. This review will therefore focus on the experimental and clinical evidence supporting neutrophils as key players in ARDS and the chemokines involved in recruiting them into the lung.


Nutrient-induced insulin resistance in human skeletal muscle.
Nutrient excess is associated with reduced insulin sensitivity (insulin resistance) and plays a central role in the pathogenesis of type 2 diabetes. Recently, free fatty acids as well as amino acids were shown to induce insulin resistance by decreasing glucose transport/phosphorylation with subsequent impairment of glycogen synthesis in human skeletal muscle. These results do not support the traditional concept of direct substrate competition with glucose for mitochondrial oxidation but indicate that the cellular mechanisms of such lipotoxicity and "proteotoxicity" might primarily affect the insulin signaling cascade. The signaling pathways involved in nutrient dependent modulation of insulin action include protein kinase C isoforms and IkappaB kinase. Therefore, pharmacological modulation of these enzymes might represent a promising target for future treatment of insulin resistance. Finally, hyperglycemia which occurs late in the insulin resistance syndrome further augments insulin resistance by mechanisms summarized as glucose toxicity. Chronic hyperglycemia might lead to inhibition of lipid oxidation and thereby to accumulation of intracellular lipid metabolites. Therefore, glucotoxicity might be in part indirectly caused by lipotoxicity (glucolipotoxicity). In conclusion, different nutrients affect common metabolic pathways and thereby induce insulin resistance in humans.


Structural evolution of differential amino acid effector regulation in plant chorismate mutases.
Chorismate mutase converts chorismate into prephenate for aromatic amino acid biosynthesis. To understand the molecular basis of allosteric regulation in the plant chorismate mutases, we analyzed the three Arabidopsis thaliana chorismate mutase isoforms (AtCM1-3) and determined the x-ray crystal structures of AtCM1 in complex with phenylalanine and tyrosine. Functional analyses show a wider range of effector control in the Arabidopsis chorismate mutases than previously reported. AtCM1 is activated by tryptophan with phenylalanine and tyrosine acting as negative effectors; however, tryptophan, cysteine, and histidine activate AtCM3. AtCM2 is a nonallosteric form. The crystal structure of AtCM1 in complex with tyrosine and phenylalanine identifies differences in the effector sites of the allosterically regulated yeast enzyme and the other two Arabidopsis isoforms. Site-directed mutagenesis of residues in the effector site reveals key features leading to differential effector regulation in these enzymes. In AtCM1, mutations of Gly-213 abolish allosteric regulation, as observed in AtCM2. A second effector site position, Gly-149 in AtCM1 and Asp-132 in AtCM3, controls amino acid effector specificity in AtCM1 and AtCM3. Comparisons of chorismate mutases from multiple plants suggest that subtle differences in the effector site are conserved in different lineages and may lead to specialized regulation of this branch point enzyme.


A Tuesday in the life of a flourisher: the role of positive emotional reactivity in optimal mental health.
Flourishing--a state of optimal mental health--has been linked to a host of benefits for the individual and society, including fewer workdays lost and the lowest incidence of chronic physical conditions. The aim of this paper was to investigate whether and how routine activities promote flourishing. The authors proposed that flourishers thrive because they capitalize on the processes featured in the broaden-and-build theory of positive emotions, specifically by experiencing greater positive emotional reactivity to pleasant events and building more resources over time. To test these hypotheses, the Day Reconstruction Method (DRM) was administered to a prescreened community sample of adults (n = 208), and they were recontacted two to three months later. Results showed that relative to those who did not flourish or were depressed, people who flourish generally responded with bigger "boosts" in positive emotions in response to everyday, pleasant events (helping, interacting, playing, learning, spiritual activity), and this greater positive emotional reactivity, over time, predicted higher levels of two facets of the cognitive resource of mindfulness. In turn, these higher levels of mindfulness were positively associated with higher levels of flourishing at the end of study, controlling for initial levels of flourishing. These results suggest that the promotion of well-being may be fueled by small, yet consequential differences in individuals' emotional experience of pleasant everyday events. Additionally, these results underscore the utility of the broaden-and-build theory in understanding the processes by which flourishing is promoted and provide support for a positive potentiation perspective.


High-Affinity Binding of Chemokine Analogs that Display Ligand Bias at the HIV-1 Coreceptor CCR5.
The chemokine receptor CCR5 is a drug target to prevent transmission of HIV/AIDS. We studied four analogs of the native chemokine regulated, on activation, normal T-cell-expressed, and secreted (RANTES) (CCL5) that have anti-HIV potencies of around 25 pM, which is more than four orders of magnitude higher than that of RANTES itself. It has been hypothesized that the ultrahigh potency of the analogs is due to their ability to bind populations of receptors not accessible to native chemokines. To test this hypothesis, we developed a homogeneous dual-color fluorescence cross-correlation spectroscopy assay for saturation- and competition-binding experiments. The fluorescence cross-correlation spectroscopy assay has the advantage that it does not rely on competition with radioactively labeled native chemokines used in conventional assays. We prepared site-specifically labeled fluorescent analogs using native chemical ligation of synthetic peptides, followed by bioorthogonal fluorescent labeling. We engineered a mammalian cell expression construct to provide fluorescently labeled CCR5, which was purified using a tandem immunoaffinity and size-exclusion chromatography approach to obtain monomeric fluorescent CCR5 in detergent solution. We found subnanomolar binding affinities for the two analogs 5P12-RANTES and 5P14-RANTES and about 20-fold reduced affinities for PSC-RANTES and 6P4-RANTES. Using homologous and heterologous competition experiments with unlabeled chemokine analogs, we conclude that the analogs all bind at the same binding site, whereas the native chemokines (RANTES and MIP-1α) fail to displace bound fluorescent analogs even at tens of micromolar concentrations. Our results can be rationalized with de novo structural models of the N-terminal tails of the synthetic chemokines that adopt a different binding mode as compared to the parent compound.


Thromboembolism during pregnancy. Risks, challenges, and recommendations.
Pregnancy is an important risk factor for venous thrombosis, and venous thromboembolism is a leading cause of preventable death in pregnancy. Diagnosis of venous thromboembolism is complicated in that the symptoms of dyspnea and lower extremity edema are relatively common complaints of pregnant patients. Physicians should maintain an appropriately high index of suspicion and request diagnostic imaging in a timely manner. Diagnosis of deep venous thrombosis with Doppler ultrasonography of the lower extremity poses no health risk to the fetus, but other radiographic studies pose a low radiation risk to the fetus. Because anticoagulant therapy poses a greater health risk to mother and fetus than does the radiation required for the diagnosis of pulmonary embolism, clinicians should aggressively pursue objective evidence of venous thromboembolism. Once the diagnosis is made, anticoagulation with intravenous unfractionated heparin or subcutaneous low-molecular-weight heparin should be used prepartum followed by warfarin therapy after delivery.


Proportion of patients needing an implantable cardioverter defibrillator on the basis of current guidelines: impact on healthcare resources in Italy and the USA. Data from the ALPHA study registry.
Implantable cardioverter defibrillators (ICD) improve survival in selected patients with left ventricular dysfunction or heart failure (HF). The objective is to estimate the number of ICD candidates and to assess the potential impact on public health expenditure in Italy and the USA. Data from 3513 consecutive patients (ALPHA study registry) were screened. A model based on international guidelines inclusion criteria and epidemiological data was used to estimate the number of eligible patients. A comparison with current ICD implant rate was done to estimate the necessary incremental rate to treat eligible patients within 5 years. Up to 54% of HF patients are estimated to be eligible for ICD implantation. An implantation policy based on guidelines would significantly increase the ICD number to 2671 implants per million inhabitants in Italy and to 4261 in the USA. An annual increment of prophylactic ICD implants of 20% in the USA and 68% in Italy would be necessary to treat all indicated patients in a 5-year timeframe. Implantable cardioverter defibrillator implantation policy based on current evidence may have significant impact on public health expenditure. Effective risk stratification may be useful in order to maximize benefit of ICD therapy and its cost-effectiveness in primary prevention.


Ultra-small Bi2S3 nanodot-doped reversible Fe(ii/iii)-based hollow mesoporous Prussian blue nanocubes for amplified tumor oxidative stress-augmented photo-/radiotherapy.
Improving the generation of reactive oxygen species (ROS) while consuming glutathione (GSH) is the main method for amplifying intracellular oxidative stress. However, in previous studies, it was normally necessary to combine two or more materials to achieve the effect of destroying the intracellular redox homeostasis. This made the preparation process relatively complicated. Herein, we designed ultra-small bismuth sulfide quantum dot (Bi2S3 QD)-doped hollow mesoporous Prussian blue (HMPB) (HMPB/Bi2S3) nanocubes for amplified tumor oxidative stress to augment photo-/radiotherapy. In addition to being photothermal materials, Prussian blue can be used as both a catalyst for the Fenton reaction and a consumer of GSH due to the multivalent state of iron. Ferrous ions (Fe(ii)) can produce toxic ROS-hydroxyl radicals (˙OH) with abundant hydrogen peroxide in the tumor cells by the Fenton reaction. Meanwhile, ferric ions (Fe(iii)) can oxidize the intracellular GSH to GSSG, thus depleting the concentration of GSH inside tumors. As a result, oxidative stress imbalance could be induced by the reversible redox property of Fe(ii/iii), thereby causing DNA damage and increasing the cell membrane permeability to realize enhanced photo-/radiotherapy. As a sensitizer for radiotherapy, ultra-small Bi2S3 QDs (3-5 nm) are doped in HMPB, thus improving the therapeutic effect by prolonging blood circulation and reducing systemic toxicity via kidney metabolism. Therefore, such a reversible HMPB/Bi2S3 nanocube is a promising therapeutic agent for amplified tumor oxidative stress to augment photo-/radiotherapy, which might show further applications in nanomedical science.


The evolving treatment landscape of chronic lymphocytic leukemia.
This review provides guidance in the rapidly changing scenario of chronic lymphocytic leukemia (CLL) treatment. New studies as well as updates of other seminal ones have been recently presented and are likely to change the management of patients with CLL in everyday clinical practice. Kinase inhibitors (e.g. ibrutinib and idelalisib) have transformed the treatment paradigm in CLL in both front-line and relapsed/refractory patients. Longer follow-up data are now available supporting the safety of ibrutinib and the continuous administration required per current label. Novel studies show the superiority of the drug alone or in combination with monoclonal antibodies compared with standard chemoimmunotherapy. The combination of venetoclax and obinutuzumab (treatment-naïve, only in United States) or rituximab (relapsed/refractory) has granted approval from the regulatory authorities in United States and Europe, based on phase 3 randomized studies. These novel chemo-free combinations allow for fixed-duration treatment and undetectable minimal residual disease. Novel targeted strategies including second and third generation BTK and PI3K inhibitors are currently under investigation and promise to further improve the CLL treatment armamentarium. The chimeric-antigen receptor (CAR) T cells are coming to the stage with promising efficacy and new challenges. A bright chemo-free era for CLL patients is just around the corner. A deep knowledge of currently available evidences is key to tailor treatment choice and optimize long-term tolerability and disease control. Fixed-duration combinations are investigated to allow treatment holidays and avoid the emergence of resistant clones under the selective pressure of continuous treatment.


Paternal mosaicism for a novel PBX1 mutation associated with recurrent perinatal death: Phenotypic expansion of the PBX1-related syndrome.
Autosomal dominant (de novo) mutations in PBX1 are known to cause congenital abnormalities of the kidney and urinary tract (CAKUT), with or without extra-renal abnormalities. Using trio exome sequencing, we identified a PBX1 p.(Arg107Trp) mutation in a deceased one-day-old neonate presenting with CAKUT, asplenia, and severe bilateral diaphragmatic thinning and eventration. Further investigation by droplet digital PCR revealed that the mutation had occurred post-zygotically in the father, with different variant allele frequencies of the mosaic PBX1 mutation in blood (10%) and sperm (20%). Interestingly, the father had subclinical hydronephrosis in childhood. With an expected recurrence risk of one in five, chorionic villus sampling and prenatal diagnosis for the PBX1 mutation identified recurrence in a subsequent pregnancy. The family opted to continue the pregnancy and the second affected sibling was stillborn at 35 weeks, presenting with similar severe bilateral diaphragmatic eventration, microsplenia, and complete sex reversal (46, XY female). This study highlights the importance of follow-up studies for presumed de novo and low-level mosaic variants and broadens the phenotypic spectrum of developmental abnormalities caused by PBX1 mutations.


Mixed fortunes: ancient expansion and recent decline in population size of a subtropical montane primate, the Arunachal macaque Macaca munzala.
Quaternary glacial oscillations are known to have caused population size fluctuations in many temperate species. Species from subtropical and tropical regions are, however, considerably less studied, despite representing most of the biodiversity hotspots in the world including many highly threatened by anthropogenic activities such as hunting. These regions, consequently, pose a significant knowledge gap in terms of how their fauna have typically responded to past climatic changes. We studied an endangered primate, the Arunachal macaque Macaca munzala, from the subtropical southern edge of the Tibetan plateau, a part of the Eastern Himalaya biodiversity hotspot, also known to be highly threatened due to rampant hunting. We employed a 534 bp-long mitochondrial DNA sequence and 22 autosomal microsatellite loci to investigate the factors that have potentially shaped the demographic history of the species. Analysing the genetic data with traditional statistical methods and advance Bayesian inferential approaches, we demonstrate a limited effect of past glacial fluctuations on the demographic history of the species before the last glacial maximum, approximately 20,000 years ago. This was, however, immediately followed by a significant population expansion possibly due to warmer climatic conditions, approximately 15,000 years ago. These changes may thus represent an apparent balance between that displayed by the relatively climatically stable tropics and those of the more severe, temperate environments of the past. This study also draws attention to the possibility that a cold-tolerant species like the Arunachal macaque, which could withstand historical climate fluctuations and grow once the climate became conducive, may actually be extremely vulnerable to anthropogenic exploitation, as is perhaps indicated by its Holocene ca. 30-fold population decline, approximately 3,500 years ago. Our study thus provides a quantitative appraisal of these demographically important events, emphasising the ability to potentially infer the occurrence of two separate historical events from contemporary genetic data.


Maternal diabetes alters transcriptional programs in the developing embryo.
Maternal diabetes is a well-known risk factor for birth defects, such as heart defects and neural tube defects. The causative molecular mechanisms in the developing embryo are currently unknown, and the pathogenesis of developmental abnormalities during diabetic pregnancy is not well understood. We hypothesized that the developmental defects are due to alterations in critical developmental pathways, possibly as a result of altered gene expression. We here report results from gene expression profiling of exposed embryos from a mouse diabetes model. In comparison to normal embryos at mid-gestation, we find significantly altered gene expression levels in diabetes-exposed embryos. Independent validation of altered expression was obtained by quantitative Real Time Polymerase Chain Reaction. Sequence motifs in the promoters of diabetes-affected genes suggest potential binding of transcription factors that are involved in responses to oxidative stress and/or to hypoxia, two conditions known to be associated with diabetic pregnancies. Functional annotation shows that a sixth of the de-regulated genes have known developmental phenotypes in mouse mutants. Over 30% of the genes we have identified encode transcription factors and chromatin modifying proteins or components of signaling pathways that impinge on transcription. Exposure to maternal diabetes during pregnancy alters transcriptional profiles in the developing embryo. The enrichment, within the set of de-regulated genes, of those encoding transcriptional regulatory molecules provides support for the hypothesis that maternal diabetes affects specific developmental programs.


[Influence of Artrofoon on the current of the metabolic syndrome in patients with rheumatoid arthritis].
This article shows observable effects of Artrofoon concerning displays a metabolic syndrome in patients with rheumatoid arthritis. Patients of the basic group received Artrofoon throughout 12 months by 4 tablets a day, thus it was possible to reach authentic decrease in level of uric acid, improvement of indicators of a lipid spectrum of blood, weight reduction, to stabilise arterial pressure. Thus, positive influence of the preparation on the basic displays of a metabolic syndrome has been noted.


Immunomodulatory properties of mesenchymal stem cell in experimental arthritis in rat and mouse models: A systematic review.
Despite recent advances in the treatment of arthritis with the development of disease-modifying antirheumatic drugs, 30% of patients still fail to respond to treatment. Given the potent anti-inflammatory and immunomodulatory properties of mesenchymal stem cells (MSC) and their ability to repair damaged cartilage, bone, and tendons, it has been proposed that MSC could be ideal for cell-based treatment of arthritis. This systematic review investigates evidence from studies on the therapeutic efficacy of MSC in rodent models of arthritis. PubMed, Embase, MEDLINE, and Wed of Science were searched to June 2015 for quantitative studies examining the outcome of treating animal models of arthritis with MSC. Inclusion criteria were as follows: administration of mesenchymal stem as a treatment approach for arthritis; animal models only; and published in English. We followed the preferred reporting items for systematic reviews and meta-analyses (PRISMA) guidelines. The literature search identified 30 studies which met the inclusion criteria. A range of MSC populations were assessed in various rodent models of arthritis. Of these, 19 demonstrated positive outcomes while 11 studies failed to demonstrate positive effects. Owing to the extensive variation in the experimental design, cells investigated and the outcome measures described in the manuscripts, no meta-analysis was possible. Furthermore, the numerical values for the primary outcome measure of clinical paw score were frequently not published in the manuscripts analyzed, as they were only illustrated in graphical form. Numerous studies have investigated the utility of a range of MSC populations in the treatment of experimental arthritis. The results obtained from these studies have been highly inconsistent, with multiple studies identifying a statistically significant improvement in arthritis scores after treatment with MSC, while other studies identified a statistically significant deterioration in arthritis scores and thirdly some studies showed no effect. Further studies using standardized protocols and outcome measures are needed to determine fully the potential of MSC populations in the treatment of experimental arthritis.


Proteomics of Galápagos Marine Iguanas Links Function of Femoral Gland Proteins to the Immune System.
Communication between individuals via molecules, termed chemosignaling, is widespread among animal and plant species. However, we lack knowledge on the specific functions of the substances involved for most systems. The femoral gland is an organ that secretes a waxy substance involved in chemical communication in lizards. Although the lipids and volatile substances secreted by the femoral glands have been investigated in several biochemical studies, the protein composition and functions of secretions remain completely unknown. Applying a proteomic approach, we provide the first attempt to comprehensively characterize the protein composition of femoral gland secretions from the Galápagos marine iguana. Using samples from several organs, the marine iguana proteome was assembled by next-generation sequencing and MS, resulting in 7513 proteins. Of these, 4305 proteins were present in the femoral gland, including keratins, small serum proteins, and fatty acid-binding proteins. Surprisingly, no proteins with discernible roles in partner recognition or inter-species communication could be identified. However, we did find several proteins with direct associations to the innate immune system, including lysozyme C, antileukoproteinase (ALP), pulmonary surfactant protein (SFTPD), and galectin (LGALS1) suggesting that the femoral glands function as an important barrier to infection. Furthermore, we report several novel anti-microbial peptides from the femoral glands that show similar action against Escherichia coli and Bacillus subtilis such as oncocin, a peptide known for its effectiveness against Gram-negative pathogens. This proteomics data set is a valuable resource for future functional protein analysis and demonstrates that femoral gland secretions also perform functions of the innate immune system.


High feline trypsin-like immunoreactivity in a cat with pancreatitis and hepatic lipidosis.
A 1.5-year-old domestic shorthair cat was examined because of vomiting and icterus. Clinicopathologic abnormalities included high alanine transaminase, alkaline phosphatase, and gamma-glutamyltransferase activities and high total bilirubin concentration. During abdominal ultrasonography, the left limb and body of the pancreas appeared hypoechoic, and a small quantity of peritoneal effusion was seen. The liver was diffusely hyperechoic, with echogenicity similar to that of the spleen, indicating hepatic lipidosis. Feline trypsin-like immunoreactivity was high, suggesting that the cat also had pancreatitis. The cat was treated with crystalloid fluids and was fed a protein-restricted diet via a percutaneous endoscopically placed gastrostomy tube. The cat's condition continued to deteriorate despite medical treatment, and it was euthanatized. Necropsy confirmed the clinical suspicion of acute pancreatitis and hepatic lipidosis. This case suggests that measurement of trypsin-like immunoreactivity may be useful in cats suspected of having pancreatitis.


Studies of natural killer cell activity and antibody-dependent cell-mediated cytotoxicity among patients with acute leukemia in complete remission.
Low natural killer (NK) cell activity against the K-562 leukemia cell line was observed in patients with acute leukemia in the early stages of remission, i.e., 2-4 months (11.3% +/- 7.95% specific target cell lysis). This parameter was found to be normal among leukemia patients after a longer time in remission (19.53% +/- 7.55%) when compared with healthy donors (18.46% +/- 12.98%). A similar pattern of activity was observed in studies of antibody-dependent cell-mediated cytolysis (ADCC) to the CEM lymphoid tumor cell line in the same group (37.58% +/- 12.4% vs. 51% +/- 6.79% specific target cell lysis). ADCC to chicken red blood cells (CRBC) and to human red blood cells (HRBC) was not significantly different from that for healthy controls at either duration of remission. Nine patients relapsed over a follow-up period of 9 months. They were found to have slightly lower NK activity (14.4% +/- 9.3%) and ADCC to CEM (41.4% +/- 8.5%) than the patients who remained in remission (17.1% +/- 6.8%; 48.7% +/- 9.7%, respectively). These data indicate a lymphocyte deficit which may persist for some time after remission has been induced, and which may be due to the effect of leukemic cell burden or the effect of aggressive chemotherapy.


Diagnostic Imaging Guidelines Implementation Study for Spinal Disorders: A Randomized Trial with Postal Follow-ups.
Implementation strategies of imaging guidelines can assist in reducing the number of radiographic examinations. This study aimed to compare the perceived need for diagnostic imaging before and after an educational intervention strategy. One hundred sixty Swiss chiropractors attending a conference were randomized to either receive a radiology workshop, reviewing appropriate indications for diagnostic imaging for adult spine disorders (n = 80), or be in a control group (CG). One group of 40 individuals dropped out from the CG due to logistic reasons. Participants in the intervention group were randomly assigned to three subgroups to evaluate the effect of an online reminder at midpoint. All participants underwent a pretest and a final test at 14-16 weeks. A posttest was administered to two subgroups at 8-10 weeks. There was no difference between baseline scores, and overall scores for the pretest and the final tests for all four groups were not significantly different. However, the subgroup provided with access to a reminder performed significantly better than the subgroup with whom they were compared (F = 4.486; df = 1 and 30; p = .043). Guideline adherence was 50.5% (95% CI, 39.1-61.8) for the intervention group and 43.7% (95% CI, 23.7-63.6) for the CG at baseline. Adherence at follow-up was lower, but mean group differences remained insignificant. Online access to specific recommendations while making a clinical decision may favorably influence the intention to either order or not order imaging studies. However, a didactic presentation alone did not appear to change the perception for the need of diagnostic imaging studies.


Draxin is involved in the proper development of the dI3 interneuron in chick spinal cord.
Generation of the appropriate types, numbers and distribution of neurons during the development of the nervous system requires the careful coordination of proliferation, differentiation, and patterning. In this work, we analyzed the roles of a repulsive axon guidance protein, draxin, on the development of chick spinal cord dI3 interneuron. draxin mRNA and/or protein were detected in the roof plate at first and then the boundary region between the ventricular and the mantle zones in chick spinal cord and dorsal basement membrane of the chick spinal cord. Overexpression of draxin caused the decreased and delayed migration of the dI3 interneuron, the reduction of progenitor cell proliferation, and abnormal localization of some ectopic progenitor-like cells in the mantle zone of the spinal cord. Our data reveal that draxin may be involved in the proper development of the dI3 interneuron in chick spinal cord.


Clinical evidence of a graft-versus-Hodgkin's-lymphoma effect after reduced-intensity allogeneic transplantation.
In patients with multiply relapsed Hodgkin's lymphoma allogeneic stem-cell transplantation has been limited by prohibitive non-relapse-related mortality rates and by a lack of definitive evidence for a therapeutic graft-versus-tumour effect. Therefore, we aimed to assess the graft-versus-tumour effect of reduced-intensity allogeneic transplantation. We undertook reduced-intensity transplantation in 49 patients with multiply relapsed Hodgkin's lymphoma, 44 (90%) of whom had progression of disease after previous autologous transplantation (median age 32 years [range 18-51], number of previous treatment courses was five [range 3-8], and time from diagnosis 4.8 years [range 0.6-4.8]). 31 patients had HLA matched donors who were related and 18 had donors who were unrelated. Median follow-up was 967 days (range 102-2232). The primary endpoints were engraftment, toxic effects, non-relapse-related mortality, incidence of graft-versus-host disease (GVHD), and the toxic effects of adjuvant donor-lymphocyte infusion. All patients engrafted. Eight of 49 (16%) had grade II-IV acute GVHD and seven (14%) had chronic GVHD before donor-lymphocyte infusion. 16 (33%) patients received donor-lymphocyte infusion from 3 months after transplantation for residual disease or progression. Six (38%) of the 16 developed grade II-IV acute GVHD and five developed chronic GVHD. Nine (56%) showed disease responses after infusion (eight complete, one partial). Non-relapse-related mortality was 16.3% at 730 days (7.2% for patients who had related donors vs 34.1% for those with unrelated donors, p=0.0206). Projected 4 year overall and progression-free survival were 55.7% and 39.0%, respectively (62.0% and 41.5% for related donors). These data show the potential for durable responses in patients who have previously had substantial treatment for Hodgkin's lymphoma. The low non-relapse-related mortality suggests the procedure could be undertaken earlier in the course of the disease.


Blood level of CD45+ platelets and development of restenosis after drug-eluting stent implantation in patients with stable coronary artery disease.
The aim of this study was to assess CD45-positive platelets (CD45+ platelets) involvement in restenosis development after drug-eluting stent (DES) implantation in patients with stable coronary artery disease (CAD). The study comprised 126 male and female patients with stable angina pectoris, who underwent elective coronary stenting with DES and follow-up angiography within 6-12 months. The patients were assigned to the group with restenosis (n = 53) or group without restenosis (n = 73) according to the follow-up angiograms. In both groups we compared the level in blood of CD45+ platelets, the clinical, laboratory and angiographic variables, which may affect the development of restenosis. We have also constructed a logit regression model for prognosis of restenosis occurrence after DES implantation. The blood count of CD45+ platelets was higher in patients with restenosis than in patients without: 0.82 % (0.58; 1.12) vs. 0.34 % (0.20; 0.68), p < 0.001, data are expressed as median (lower quartile; upper quartile). By binary comparisons of more than 35 different clinical, laboratory and angiographic variables we identified 8 significant risk factors for the development of stent restenosis after DES. In order to define the risk of the development of restenosis, we have built a logit regression model. The resulting logit regression equation included the level of CD45+ platelets, the neutrophil to lymphocyte ratio (NLR), small diameter arteries stenting and the number of simultaneously implanted stents in one patient. Receiver operating characteristic (ROC) curve analysis has demonstrated the high prognostic value of the resulting logit regression equation with an area under the curve (AUC) of 0.91 % (p < 0.001). The acquired data indicate the presence of a close relationship between circulating CD45+ platelets and restenosis development after DES implantation in patients with stable CAD.


Experimental edema disease of swine (E. coli enterotoxemia). I. Dectection and preparation of an active principle.
Freeze-thaw lysates prepared from strains of Escherichia coli belonging to serogroups O138, O139, and O141 contained a principle (edema disease principle) which induced edema disease in swine. All freeze-thaw lysates contained endotoxic activity that tended to obscure the edema disease syndrome and methods were developed to reduce such activity. Freeze-thaw lysates prepared from E. coli O139 induced the most characteristic edema disease syndrome. Partially purified edema disease principle prepared from O139 freeze-thaw lysates by sequential precipitation with ammonium sulphate and streptomycin sulphate had increased specific activity with markedly reduced endotoxic activity. This material was insoluble at acidic pH but readily soluble at alkaline pH. The effective molecular weight of edema disease principle, based on retention and filtration properties of diaflo membranes, appeared to be greater than 50,000 and less than 100,000. The biological activity of edema disease principle was thermolabile. Sodium deoxycholate treatment of edema disease principle further reduced endotoxic activity. A thermolabile, ammonium sulphate precipitable material was prepared from E. coli O139 that induced a predictable syndrome which resembled edema disease clinically and pathologically following intravenous inoculation in pigs.


[Morphological studies of intraepithelial carcinomas of the cervix uteri (author's transl)].
A morphological study at light and electron microscopy, of 61 cases of intraepithelial epithelioma allowed the authors to distinguish two types of carcinoma: undifferentiated intraepithelial epithelioma arising from the exo-endocervical junction, probably by proliferation of vestigial cells which derived from persistent sinusal remnants; differentiated intraepithelial epithelioma developing upon the mature exocervical epithelium modified by inflammatory processes. These two types of "in situ" epitheliomas have different evolution. The undifferentiated form, supported by an uninterrupted membrane consisting of imbricated reticulin fibres has a slow evolution. The differentiated form, often multifocal, supported by a basal membrane frequently disrupted, has a rapid evolution. These morphological studies allowed the authors to refuse the theory of metaplasia from subcylindrical reserve cells in the genesis of intraepithelial epitheliomas of the cervix.


"We literally worked in parking lots, cars, garages, and separately set up party tents": qualitative study on the experiences of GPs in the frame of the SARS-CoV-2 pandemic in Austria.
Primary care is internationally recognised as one of the cornerstones of health care. During the COVID-19 pandemic, primary care physicians were assigned a variety of tasks and thus made a significant contribution to a country's pandemic response. They were expected to perform a variety of tasks, such as diagnosing and treating people with COVID-19, maintaining health care for all other patients, as well as several public health tasks, such as diagnostic testing and vaccination, protecting patients and staff from infection, and serving as community trusted persons. In Austria, there are no structured levels of care, no definition of the role of the general practitioner during a pandemic is given, and no specific support structures are present. The aim of this study was to assess the views and experiences of primary care physicians regarding supportive and hindering factors for pandemic preparedness in Austria. Qualitative study using semi-structured interviews. A total of 30 general practitioners were interviewed, with particular attention to an equitable distribution in small, medium and large primary care facilities. Qualitative content analysis was performed. Interviewees described a wide range of infection control, organisational and communication measures that they had implemented. They made changes to practise equipment, found makeshift solutions when supplies were scarce, and established communication and information pathways when official communication lines were inadequate. General practitioners took on essential tasks and showed a high level of understanding of their role in the pandemic response. This was achieved mainly at an informal level and with high personal commitment. Their functioning in the absence of structural regulations and support shows that they had a clear intrinsic understanding of their responsibilities. To ensure reliability and sustainability and to reduce their burden, it will be necessary to clarify the role and tasks of a general practitioner and to provide the necessary support. This concerns both infrastructural support and communication and information strategies. As part of the reform to strengthen primary care, primary care needs to be seen, valued and involved in decision-making processes.