Aerial monitoring of marine waterfowl in the Alaskan Beaufort Sea.
The purpose of this study was to design and test a monitoring protocol for marine waterfowl in the central Alaskan Beaufort Sea. The study provides an important case-study of how a long-term monitoring program may be affected by unanticipated human disturbances. Because of its overwhelming and widespread abundance, relatively sedentary behavior, ease in counting, and the extensive historical database, the long-tailed duck (Clangula hyemalis) was selected as the focal species. Two null hypotheses were formulated concerning potential changes in the numbers and distribution of long-tailed ducks in relation to disturbance in an industrial study area, compared to a reference study area located about 50 km to the east. A 9-year historical database (1977-1984, 1989) of long-tailed duck densities and other important data recorded during systematic aerial surveys was analyzed retrospectively using multiple regression techniques. The retrospective analyses determined which of several predictor variables recorded were significantly related to long-tailed duck density. Separate analyses were conducted for two periods: (1) the overall period when long-tailed ducks were present in the lagoon study areas, and (2) the shorter adult male molt period. The results of the two analyses indicated that 57% and 68%, respectively, of the total variation in long-tailed duck density during the two periods could be explained by variables recorded during the surveys. Predictor variables representing habitat, day of the year, time of day, amount of ice, and wave height recorded on-transect during surveys were most closely associated with long-tailed duck density. Measurement error during the surveys, and influences outside the study area such as nesting success in tundra habitats and mortality during migration and in over-wintering areas likely also had strong influences on the results, but these factors were not measurable in our study. Based on results of the retrospective analyses, a long-term monitoring protocol consisting of a program of systematic aerial surveys and an analyses of variance and covariance (ANOVA and ANCOVA) statistical procedure was designed and initially tested in 1990 and 1991. This 2-year testing phase resulted in several revisions to the monitoring protocol. Refinements were made to the original sampling procedures, to the survey schedule, and to the recommended statistical analysis procedures. Results of the ANOVA and ANCOVA indicated that there was no evidence of a change in long-tailed duck densities that could be attributable to disturbance (from any source) in the industrial study area relative to a reference area with no industrial development. Other analyses indicated that the sampling and analysis procedures would be adequate to detect long-term trends in long-tailed duck density and localized disturbance effects, but that the monitoring program should be continued well beyond two years to detect statistically significant changes. As a result, additional aerial surveys of both study areas were conducted again during 1999-2001. Results of the revised ANOVA and ANCOVA of the 1990-1991 and 1999-2001 survey data indicated that the density of long-tailed ducks had significantly declined in coastal lagoons along the central Alaskan Beaufort Sea coast during the study period. In addition, disturbances throughout the barrier island-lagoon systems used by these ducks, including both the industrial and the reference study areas, had significantly increased over the same period. However, because unanticipated disturbances from a variety of anthropogenic sources, and not just industry sources, increased in both study areas, the reference study area was not an effective statistical control. As a result, the decline in long-tailed duck density in both study areas was not attributable to industry-related activities. Although the monitoring protocol described here is an effective method to detect statistically significant changes in long-tailed duck distribution and abundance in the nearshore Alaskan Beaufort Sea, many more years of sampling would be necessary to attribute observed changes to industry-related disturbances.


Tracking the temporal variation of COVID-19 surges through wastewater-based epidemiology during the peak of the pandemic: A six-month long study in Charlotte, North Carolina.
The global spread of SARS-CoV-2 has continued to be a serious concern after WHO declared the virus to be the causative agent of the coronavirus disease 2019 (COVID-19) a global pandemic. Monitoring of wastewater is a useful tool for assessing community prevalence given that fecal shedding of SARS-CoV-2 occurs in high concentrations by infected individuals, regardless of whether they are asymptomatic or symptomatic. Using tools that are part of wastewater-based epidemiology (WBE) approach, combined with molecular analyses, wastewater monitoring becomes a key piece of information used to assess trends and quantify the scale and dynamics of COVID-19 infection in a specific community, municipality, or area of service. This study investigates a six-month long SARS-CoV-2 RNA quantification in influent wastewater from four municipal wastewater treatment plants (WWTP) serving the Charlotte region of North Carolina (NC) using both RT-qPCR and RT-ddPCR platforms. Influent wastewater was analyzed for the nucleocapsid (N) genes N1 and N2. Both RT-qPCR and RT-ddPCR performed well for detection and quantification of SARS-CoV-2 using the N1 target, while for the N2 target RT-ddPCR was more sensitive. SARS-CoV-2 concentration ranged from 103 to 105 copies/L for all four plants. Both RT-qPCR and RT-ddPCR showed a significant positive correlation between SARS-CoV-2 concentrations and the 7-day rolling average of clinically reported COVID-19 cases when lagging 5 to 12 days (ρ = 0.52-0.92, p < 0.001-0.02). A major finding of this study is that RT-qPCR and RT-ddPCR generated SARS-CoV-2 data that was positively correlated (ρ = 0.569, p < 0.0001) and can be successfully used to monitor SARS-CoV-2 signals across the WWTP of different sizes and metropolitan service functions without significant anomalies.


Tumor Suppressor PTPRJ Is a Target of miR-155 in Colorectal Cancer.
PTPRJ is known for its antiproliferative role. Loss of heterozygosity (LOH) of PTPRJ has frequently been observed in various human cancers including colorectal cancer (CRC), lung cancer, and breast cancer. However, the function and mechanism of PTPRJ in CRC are not well understood. At the present study, we show that ectopic expression of PTPRJ inhibits cell growth, migration, and invasiveness in CRC cell line HCT116. Moreover, PTPRJ inhibits the tumorigenecity of HCT116 in a xenograft tumor model. MiR-155, the well-known oncomiR in CRC, is identified as an upstream factor of PTPRJ. MiR-155 directly binds to the 3' untranslated region of PTPRJ mRNA and suppresses the mRNA and protein levels of PTPRJ. Furthermore, the growth-promoting and AKT signaling activation effect of miR-155 was abrogated by PTPRJ overexpression, and vice versa. Our study reveals the crucial role of miR-155/PTPRJ/AKT axis in proliferation and migration of CRC cells and suggests a therapeutic potential of PTPRJ. J. Cell. Biochem. 118: 3391-3400, 2017. © 2017 Wiley Periodicals, Inc.


Immunohistochemical distribution of beta-protein kinase C in rat hippocampus determined with an antibody against a synthetic peptide sequence.
An antibody directed against a synthetic peptide sequence specific for the beta-subtype of protein kinase C (PKC) was used to determine the distribution of beta-PKC in rat hippocampus by immunocytochemistry. PKC was distributed primarily in the stratum oriens and radiatum of the CA1 region. Positive staining cell bodies were only observed after colchicine treatment in pyramidal cells (CA2-CA4) and granule cells of the dentate gyrus. The discrete localization of various subtypes of PKC should provide clues to their functions.


Clinical characteristics of patients infected with hepatitis B virus genotypes A, B, and C.
The aim of the present study was to evaluate the clinical characteristics at first hospital consultation, according to the genotype of hepatitis B virus (HBV), in patients with chronic liver diseases and positivity for hepatitis B surface antigen (HBsAg) in metropolitan Tokyo. The subjects consisted of 1077 patients with chronic liver diseases who were HBsAg-positive. HBV genotype was determined by an enzyme-linked immunosorbent assay (ELISA), using PreS2 monoclonal antibody, which is specific for HBV genotypes in the PreS2 region. The proportion of patients with genotype A was 2% (20 patients), genotype, B 9% (101 patients), and genotype C, 88% (945 patients), while 11 patients (1.0%) had other genotypes, including 2 (0.2%) each with genotypes D and F, and 7 (0.6%) that were untypeable. Patients with genotype A were significantly (P = 0.049) more likely to be positive for Hepatitis B envelope antigen (HBeAg) at the first consultation compared with those with genotype B. Patients with genotype B were significantly more likely to be HBeAg-negative at the first consultation compared with those with genotype A (P = 0.049) and genotype C (P = 0.001), significantly more likely to show minimal hepatic fibrosis (P = 0.003), and least likely to develop liver cirrhosis (P = 0.005). Patients with genotype C were likely to be HBeAg-positive (P = 0.001) and to have a positive family history of HBV infection (P = 0.0002). The clinical features of patients with HBV infection in metropolitan Tokyo varied depending on the HBV genotype.


Can the corneal tissue be used as a wound dressing material for skin injuries?
Currently, the application field of corneal tissue is corneal transplantation from animal to human or from human to human. Although the cornea and skin emerge from the same embriyological layer, ectoderm, and the cornea has a very high capacity, no research has been conducted to reveal the effect of the cornea in skin wound healing. There is no commercial product available in the hospital pharmacy that contains a ready-to-use, hygienic, wound dressing material manufactured from cornea for both ocular or extraocular wounds. In this article, the feasibility of corneal tissue as a candidate for disposable wound dressing material for skin injuries are examined and compared with the closest alternative, amnion membrane.


Design, synthesis and activity of benzothiazole-based inhibitors of NO production in LPS-activated macrophages.
Series of benzothiazoles were synthesized and evaluated their inhibitory activities for NO production in lipopolysaccharide-activated macrophages. The most potent compound was the indole-containing benzothiazole 3c with 4.18μM of IC(50). The mechanistic study suggested that benzothiazoles inhibited NO production by the suppression of iNOS protein and mRNA expression. They also suppressed the expression of COX-2 through the NF-κB inactivation.


Conjugated Polymers with Oligoethylene Glycol Side Chains for Improved Photocatalytic Hydrogen Evolution.
Conjugated polymers are emerging as promising organic photocatalysts for hydrogen evolution from water. However, it is still very challenging for conjugated polymers to realize highly efficient photocatalytic hydrogen evolution. Herein, we demonstrate an efficient strategy of hydrophilic side chain functionalization to boost the hydrogen evolution rates of conjugated polymers. By functionalizing conjugated polymers with hydrophilic oligo (ethylene glycol) monomethyl ether (OEG) side chains, a 90-fold improvement in hydrogen evolution rate has been achieved than that of alkyl-functionalized conjugated polymer. It is found that the OEG side chains interact robustly with Pt co-catalysts, resulting in more efficient charge transfer. Moreover, OEG side chains in conjugated polymers can adsorb H+ from water, resulting in significantly lowered energy levels on the surfaces of conjugated polymers, which enables cascade energy levels and enhances charge separation and photocatalytic performance. Our results indicate that rational side-chain engineering could facilitate the design of improved organic photocatalysts for hydrogen evolution.


Polyadenylate-protein complexes in resting and growing 3T3 cells.
The properties of the ribonuclease resistant cytoplasmic ribonucleoprotein particles were studied in contact-inhibited and serum induced proliferating 3T3 cells. The RNP particles were fractionated by oligo (dT)-cellulose chromatography and banded in CsSO4 gradients. The main RNP fraction, eluted with 25% formamide, contained the major ribonuclease resistant RNA sequences in both resting and growing cells. The protein component of this fraction had a molecular weight of about 72,000 in contact-inhibited cells and 81,000 in serum induced cells.


Hypoxia-ischemia induces transforming growth factor beta 1 mRNA in the infant rat brain.
Transforming growth factor beta 1 (TGF beta 1) mRNA expression was examined after hypoxia-ischemia in rat brains using in situ hybridization. Twenty-one-day-old Wistar rats had unilateral ligation of the right carotid artery followed by either 15 or 90 min inhalational hypoxia. Fifteen min of hypoxia resulted in moderate damage with selective neuronal loss in cortical layer 3 and in the hippocampus of the ligated hemisphere. Seventy-two hours after hypoxia TGF beta 1 expression was markedly increased above control levels in those sites. Levels were normal after 120 h. Ninety min of hypoxia led to an infarction of the lateral cerebral cortex and hippocampus of the ligated hemisphere. One hour after hypoxia TGF beta 1 mRNA was expressed in the hippocampus of the damaged side. Seventy-two and 120 h after hypoxia, expressing cells were found throughout the cerebral cortex, piriform cortex, striatum, thalamus and hippocampus of the infarcted side. These data show that TGF beta 1 mRNA expression is induced after a hypoxic-ischemic insult in the brain. TGF beta 1 may be involved in post-asphyxial repair mechanisms.


Generation of a human haploid neural stem cell line for genome-wide genetic screening.
Haploid embryonic stem cells (haESCs) have been established in many species. Differentiated haploid cell line types in mammals are lacking due to spontaneous diploidization during differentiation that compromises lineage-specific screens. To derive human haploid neural stem cells (haNSCs) to carry out lineage-specific screens. Human haNSCs were differentiated from human extended haESCs with the help of Y27632 (ROCK signaling pathway inhibitor) and a series of cytokines to reduce diploidization. Neuronal differentiation of haNSCs was performed to examine their neural differentiation potency. Global gene expression analysis was con-ducted to compare haNSCs with diploid NSCs and haESCs. Fluorescence activated cell sorting was performed to assess the diploidization rate of extended haESCs and haNSCs. Genetic manipulation and screening were utilized to evaluate the significance of human haNSCs as genetic screening tools. Human haESCs in extended pluripotent culture medium showed more compact and smaller colonies, a higher efficiency in neural differentiation, a higher cell survival ratio and higher stability in haploidy maintenance. These characteristics effectively facilitated the derivation of human haNSCs. These human haNSCs can be generated by differentiation and maintain haploidy and multipotency to neurons and glia in the long term in vitro. After PiggyBac transfection, there were multiple insertion sites in the human haNSCs' genome, and the insertion sites were evenly spread across all chromosomes. In addition, after the cells were treated with manganese, we were able to generate a list of manganese-induced toxicity genes, demonstrating their utility as genetic screening tools. This is the first report of a generated human haploid somatic cell line with a complete genome, proliferative ability and neural differentiation potential that provides cell resources for recessive inheritance and drug targeted screening.


Feedback on patient progress and clinical support tools for therapists: improved outcome for patients at risk of treatment failure in psychosomatic in-patient therapy under the conditions of routine practice.
Although psychosomatic in-patient treatment is effective, 5-10% of the patients deteriorate. Providing patient progress feedback and clinical support tools to therapists improves the outcome for patients at risk of deterioration in counseling, outpatient psychotherapy, and substance abuse treatment. This study investigated the effects of feedback on psychosomatically treated in-patients at risk of treatment failure. At intake, all patients of two psychosomatic clinics were randomized either into the experimental group or the treatment-as-usual control group. Both groups were tracked weekly with the "Outcome Questionnaire" (OQ-45) measuring patient progress and with the clinical support tool "Assessment of Signal Cases" (ASC). Therapists received feedback from both instruments for all their experimental group patients. "Patients at risk" were defined as patients who deviated from expected recovery curves by at least one standard deviation. Of 252 patients, 43 patients were at risk: 23 belonged to the experimental group, 20 to the control group. The feedback effect was analyzed using a level-2-model for discontinuous change, effect size (d), reliable change index (RCI), and odds ratio for reliable deterioration. For patients at risk, the experimental group showed an improved outcome on the OQ-45 total scale compared to the control group (p<0.05, d=0.54). By providing feedback, the rate of reliably deteriorated patients at risk was reduced from 25.0% (control group) to 8.7% (experimental group) - odds ratio=0.29. All reliably improved patients at risk belonged to the experimental group. Feedback improves the outcome of patients at risk undergoing psychosomatic in-patient treatment.


Characteristics of Sepsis-2 septic shock patients failing to satisfy the Sepsis-3 septic shock definition: an analysis of real-time collected data.
Baseline characteristics and disease severity of patients with septic shock according to the new Sepsis-3 definition may differ from patients that only comply with the Sepsis-2 definition. We conducted a retrospective cohort study on the ICU of a Belgian tertiary care facility to seek out differences between these two patient groups and to identify variables associated with no longer satisfying the latest definition of septic shock. Of 1198 patients with septic shock according to the Sepsis-2 consensus definition, 233 (19.4%) did not have septic shock according to the Sepsis-3 shock definition. These patients more often had medical admission reasons and a respiratory infection as cause for the septic shock. They less often had surgery on admission and were less likely to have chronic liver disease (5.6% vs 16.2%, absolute difference 10.6% (95% CI 6.4-14.1%). Patients with septic shock only according to the old definition had significant lower APACHE II and SOFA scores and lower hospital mortality (31.6% vs 55.3%, p < 0.001). In a multivariate analysis, following variables were associated with Sepsis-2 shock patients no longer being defined as such by the Sepsis-3 definition: respiratory infection (OR 1.485 (95% CI 1.56-2.089), p = 0.023), a medical admission reason (OR 1.977 (95% CI 1.396-2.800) and chronic liver disease (OR 0.345 (95% CI 0.181-0.660), p < 0.001). One in five patients with septic shock according to the Sepsis-2 consensus definition is no longer considered as such when the Sepsis-3 shock criteria are applied. A medical admission reason, a respiratory infection and absence of chronic liver disease are independently associated with no longer being identified as having septic shock by the Sepsis-3 criteria.


Ventricular refractory periods in relation to rate and test-site VT intervals in anesthetized and conscious dogs: a canine model for conscious state measurements.
The functional relationship of ventricular effective refractory period (ERP) with basic cycle length (BCL) of stimulated ventricular depolarization and VT intervals of test-site unipolar ventricular electrograms were studied in five conscious and eight anesthetized (sodium pentobarbital, 30 mg/kg) open-chested dogs. The range of BCLs studied was 300 to 1000 ms, achieved through ventricular stimulation following chemically-induced complete AV block. In the conscious animal model developed for this study, two-to-four ventricular electrode leads were exteriorized and the ventricular rate was maintained by an implanted programmable VVI pacemaker. In all animals studied, the BCL-ERP relation was closely represented by the empirical equation ERP = A-B.Exp(-k.BCL), and the VT-ERP relation over the same range of BCLs was linear: ERP = C + D.VT. The correlation coefficients were in the range of 0.991 to 0.999. The mean values of the parameters in the above equations determined by the appropriate non-linear or linear regression analysis showed significant differences between the two groups of animals studied. In three conscious animals a strong linear correlation between the test-site VT intervals and simultaneously measured QT intervals measured from the lead II surface electrogram was demonstrated (r = 0.993 to 0.998). For a fixed site of stimulation, the morphology of ventricular depolarization complexes as well as the corresponding T-waves remained essentially unaltered with BCL for both myocardial and surface electrograms. The possible applications of developed canine model and the results of the present study include: (1) the study of the rate-dependent effects of cardioactive drugs on ventricular electrophysiology and, (2) the improved design of electronic refractory periods of rate programmable pacemakers.


Neurogenic bladder dysfunction.
The neurophysiology of lower urinary tract function is followed by a discussion of the causes, evaluation, and management of neurogenic bladder dysfunction in children. All forms of injured bladders require early assessment and proper treatment, the basis for which is provided. Clues to what the pediatrician can look for and care for are presented.


[Compression of the ulnar nerve at Guyon's canal caused by a pseudoaneurysm of the ulnar artery following trauma].
We repor there on a 16-year-old patient who presented with pain and swelling in the hypothenar eminence as well as loss of sensibility in the fingers of the region innervated by the ulnar nerve; this happened 2-3 weeks after an injury by a glass splinter in his proximal palm. A pseudoaneurysm could be verified by duplex sonography. The patient wished to avoid any graft for arterial bridging for religious reasons. On the basis of an the Allen test preoperatively and the intraoperative findings, an adequate blood supply of the finger by the radial artery was expected. Thus, in respect to the patients wish, the aneurysm was resected without bridging. The patient recovered perfectly. 4 years later, an MR-angiography showed the deep and superficial transverse palmar arc to be supplied by a voluminous radial artery. The ulnar finger arteries originated from the deep arc, the radial finger arteries from the superficial arc. In this paper, the criteria pro and contra grafting the ulnar atery at Guyons's canal will be discussed.


Percutaneous High-Energy Microwave Ablation for the Treatment of Pulmonary Tumors: A Retrospective Single-Center Experience.
To evaluate the safety and efficacy of percutaneous high-energy microwave ablation (MWA) for the treatment for pulmonary tumors. A retrospective review was undertaken of 44 patients (21 men, 23 women; median age, 66 y; range, 17-89 y) who underwent 62 sessions of high-energy MWA for 87 pulmonary tumors at a single tertiary referral center between June 2012 and June 2014. Primary tumor origin was sarcoma (n = 23), colorectal (n = 16), lung (n = 2), esophageal (n = 1), breast (n = 1), and bladder (n = 1). Median tumor size was 12 mm (range, 6-45 mm). Technical success was recorded contemporaneously, complication rate at 30 days was recorded prospectively, and technique effectiveness was assessed by longitudinal follow-up CT scan. Primary technical success was achieved in 94% of ablation sessions. The median follow-up interval was 15 months (range, 6.2-29.5 mo) during which time local tumor progression was observed in two of 87 tumors (technique effectiveness 98%). Pneumothorax requiring chest tube insertion occurred in 19%; delayed pneumothorax occurred in four patients. No hemoptysis, infection, or other complications were recorded. High-energy MWA is safe and effective for the destruction of lung tumors.


Effects of nitrogen and phosphorus concentrations on the bioaccumulation of polybrominated diphenyl ethers by Prorocentrum donghaiense.
The growth, cellular total lipids, bioaccumulation amount, and bioaccumulation factors (BAFs) of 2,4,4'-tribromodiphenyl ether (BDE28), 2,2',4,4'-tetrabromodiphenyl ether (BDE47), and 2,2',4,4',5-pentabromodiphenyl ether (BDE99) in a semi-continuous culture of Prorocentrum donghaiense were studied in relation to nitrate (0, 128, and 512 micromol/L) and phosphate (0, 8, and 32 micromol/L) concentrations. The BDE28, BDE47, and BDE99 content per cell under 0 micromol N/L were 3.77 x 10(-6), 3.95 x 10(-6), and 4.32 x 10-6 ng/cell, respectively, which were significantly higher than those under 128 and 512 micromol N/L. A nearly 5-fold increase in polybrominated diphenyl ether (PBDE) content per algal cell was found between 0 and 8 micromol P/L and between 8 and 32 micromol P/L. With increasing N and P concentrations, the PBDE content per volume of algal culture and the accumulation percentage of available PBDEs declined slightly. The BAFs for the PBDEs based on lipids showed that the logBAF(lip) under 0 micromol N/L was higher than those under 128 and 512 micromol N/L. The logBAF(lip) under 0 micromol P/L was higher than that under 8 micromol P/L but lower than that under 32 micromol P/L. Correlation analysis indicated a significant negative correlation between nutrient concentration and cellular total lipids, as well as the PBDE content per cell. The results indicate that different N and P concentrations change the total lipids content of P. donghaiense, thereby resulting in varying PBDE accumulation.


Impact of diabetes on immature platelets fraction and its relationship with platelet reactivity in patients receiving dual antiplatelet therapy.
Contrasting data have been reported so far on the role of reticulated platelets in suboptimal response to antiplatelet therapies. In particular, still unexplored is whether they may contribute to explain the higher risk of thrombotic complications observed in diabetic patients. Aim of the present study was to evaluate the impact of diabetes on the levels of reticulated platelets and its relationship with high residual on-treatment platelet reactivity (HRPR) in patients receiving dual antiplatelet therapy. In patients treated with ASA (100-160 mg) and clopidogrel (75 mg daily) or ticagrelor (90 mg twice a day) platelet reactivity and the reticulated platelets fraction (immature platelets fraction, IPF) were assessed at 30-90 days post-discharge for an acute coronary syndrome or elective PCI. Aggregation was assessed by multiple-electrode aggregometry. We included 386 patients, 158 (40.9 %) diabetics. The percentage of IPF was similar in diabetic and non diabetic patients, both at baseline (3.5 ± 2.5 vs 3.6 ± 2.7 %, p = 0.91) and at 30-90 days re-assessment (3.3 ± 2.1 vs 3.5 ± 2.5 %, p = 0.30), with diabetes not emerging as an independent predictor of IPF above III tertile (adjusted OR [95 %CI] = 0.58 [0.30-1.09], p = 0.10). Diabetic patients displayed an enhanced platelet reactivity and a higher rate of HRPR with ADP antagonists (32.8 vs 22.5 %, p = 0.009). However, no association was found between the percentage of IPF and platelet function (r = -0.004; p = 0.95 for ASPI test, r = -0.04; p = 0.59 for ADP-mediated aggregation), or the rate of HRPR for ADP antagonsist across IPF tertiles. Results were similar for diabetics both receiving clopidogrel and ticagrelor. Diabetic patients display a higher platelet reactivity and suboptimal response to ADP-antagonists. However, the rate of reticulated platelets is neither influenced by diabetic status nor associated with an increased platelet reactivity among diabetic patients receiving dual antiplatelet therapy for a recent acute coronary syndrome or PCI.


Impact of crossbridge structure on peptidoglycan crosslinking: A synthetic stem peptide approach.
The bacterial cell wall, whose main component is peptidoglycan (PG), provides cellular rigidity and prevents lysis from osmotic pressure. Moreover, the cell wall is the main interface between the external environment and internal cellular components. Given its essentiality, many antibiotics target enzymes related to the biosynthesis of cell wall. Of these enzymes, transpeptidases (TPs) are central to proper cell wall assembly and their inactivation is the mechanism of action of many antibiotics including β-lactams. TPs are responsible for stitching together strands of PG to make the crosslinked meshwork of the cell wall. This chapter focuses on the use of solid-phase peptide synthesis to build PG analogs that become site-selectively incorporated into the cell wall of live bacterial cells. This method allows for the design of fluorescent handles on PG probes that will enable the interrogation of substrate preferences of TPs (e.g., amidation at the glutamic acid residue, crossbridge presence) by analyzing the level of probe incorporation within the native cell wall of live bacterial cells.


Repair of femoral capital physeal fractures with 7.0 mm cannulated screws in cattle: 20 cases (1988-2002).
To evaluate long-term outcome of repair of femoral capital physeal fractures with 70 mm cannulated screws in juvenile bulls that weighed >350 kg (770 lb). Retrospective study. 20 bulls. Medical records of affected cattle were identified and reviewed. Follow-up information was obtained by means of a written questionnaire, 1 to 15 years after discharge, from the veterinarians who supervised the postoperative care at bull stud facilities. Mean age and body weight at time of repair were 19 months (range, 11 to 27 months) and 513 kg (1128.6 lb; range, 364 to 720 kg [800.8 to 1584 lb]), respectively. Surgical repair was performed with 70 mm cannulated screws. A second surgery to replace malpositioned implants was required in 1 bull. Fourteen of the 20 bulls were considered serviceable for semen collection after surgical repair, and persistent lameness was evident in 11 of the 14. Mean duration for bulls to become serviceable was 5.5 months (range, 2 to 11 months). Age, weight, duration of injury at the time of repair, and degree of reduction did not have a significant effect on whether surgical repair was successful. Six bulls remained severely lame and were euthanatized. Results indicated that surgical repair of femoral capital physeal fractures in adolescent bulls with 70 mm cannulated screws placed in lag fashion has a good prognosis for long-term function in a semen collection facility.


Vestibular impairments in episodic ataxia type 2.
Episodic ataxia type 2 (EA2) can present diverse ocular motor abnormalities, but few studies have systematically evaluated vestibular function during the interictal periods. This study aimed to determine vestibular impairments in patients with EA2 during the interictal periods. We recruited 17 patients with genetically confirmed EA2 (10 men, age range = 16-85 years, median = 32 years). We systematically evaluated the vestibular function by measuring the semicircular canals (SCCs) function with bithermal caloric tests, rotatory chair test, and video head impulse test (vHIT), and the otolith function with subjective visual vertical (SVV) tilt and variability, and cervical and ocular vestibular-evoked myogenic potentials (VEMPs). Patients with EA2 commonly showed abnormal VOR responses at least for one SCC with high-acceleration, high-frequency head impulses (14/16, 88%), and impaired visual-vestibular interaction (7/12, 58%). In response to low acceleration and frequency stimuli, the VOR gains were generally normal. The majority of EA2 patients had impairments in at least one of the otolith function tests (13/16, 81%): SVV tilt or variability (7/14, 50%), oVEMP (8/15, 53%), and cVEMP (4/16, 25%). Vestibular impairments are common in EA2 even during the interictal periods. Selective decrease in the VOR responses during higher acceleration stimuli along with impaired visual-vestibular interaction and otolith function suggests degeneration of the vestibulocerebellum or vestibular nuclei.


Serum deoxyribonuclease I activity can be a useful diagnostic marker for the early diagnosis of unstable angina pectoris or non-ST-segment elevation myocardial infarction.
Recently, serum deoxyribonuclease I (DNase I) activity has been highlighted as a potential diagnostic marker for transient myocardial ischemia. To evaluate whether serum DNase I activity can be a useful biomarker for diagnosing unstable angina pectoris (UAP) or non-ST-segment elevation myocardial infarction (NSTEMI), we investigated serial changes in DNase I levels after chest pain in UAP and NSTEMI (UAP/NSTEMI) patients. Thirty-three and ten patients classified into the UAP/NSTEMI and the chest pain syndrome (CPS) group, respectively, were enrolled. The serum DNase I activity levels within 3h after chest pain and the absolute median value of percentage differences in serum DNase I activity levels from admission to 3h after hospitalization in the UAP/NSTEMI patients was significantly higher than those in the CPS patients. We evaluated the patients to show positive results for DNase I activity if the serum levels or percentage differences exceeded the corresponding cut-off values. The sensitivity and specificity of DNase I within 6h after chest pain in the UAP/NSTEMI patients without elevated levels of cardiac troponin T and the MB isoenzyme of creatine kinase were 89% and 88%, respectively. Serum DNase I activity can be a useful marker for the early diagnosis of UAP/NSTEMI after the onset of chest pain, irrespective of the evidence of myocardial injury.


(-)-Epigallocatechin gallate-loaded polycaprolactone scaffolds fabricated using a 3D integrated moulding method alleviate immune stress and induce neurogenesis.
In peripheral neuropathy, the underlying mechanisms of nerve and muscle degeneration include chronic inflammation and oxidative stress in fibrotic tissues. (-)-Epigallocatechin gallate (EGCG) is a major, active component in green tea and may scavenge free radical oxygen and attenuate inflammation. Conservative treatments such as steroid injection only deal with early, asymptomatic, peripheral neuropathy. In contrast, neurolysis and nerve conduit implantation work effectively for treating advanced stages. An EGCG-loaded polycaprolactone (PCL) porous scaffold was fabricated using an integrated moulding method. We evaluated proliferative, oxidative and inflammatory activity of rat Schwann cells (RSCs) and rat skeletal muscle cells (RSMCs) cultured on different scaffolds in vitro. In a rat radiation injury model, we assessed the morphological, electrophysiological and functional performance of regenerated sciatic nerves and gastrocnemius muscles, as well as oxidative stress and inflammation state. RSCs and RSMCs exhibited higher proliferative, anti-oxidant and anti-inflammatory states in an EGCG/PCL scaffold. In vivo studies showed improved nerve and muscle recovery in the EGCG/PCL group, with increased nerve myelination and muscle fibre proliferation and reduced macrophage infiltration, lipid peroxidation, inflammation and oxidative stress indicators. The EGCG-modified PCL porous nerve scaffold alleviates cellular oxidative stress and repairs peripheral nerve and muscle structure in rats. It attenuates oxidative stress and inflammation in vivo and may provide further insights into peripheral nerve repair in the future.


[Orgotein iontophoresis in the therapy of induratio penis plastica].
In objective terms the results of medical and physical treatment of Peyronie's disease are still indubitably disappointing. However personal experience has shown that medicated ionophoresis using orgotein does at least eliminate or reduce the pain in Peyronie's disease.


Termination of paroxysmal supraventricular tachycardia by intravenous adenosine in a child.
A fit 6-year-old child developed paroxysmal supraventricular tachycardia at the end of an operation for adenotonsillectomy. Sinus rhythm was rapidly restored with intravenous adenosine.


First genetic characterization of Fasciola hepatica in Argentina by nuclear and mitochondrial gene markers.
Fasciola hepatica is a trematode showing genetic variation among isolates from different regions of the world. The objective of this work was to characterize for the first time F. hepatica isolates circulating in different regions of Argentina. Twenty-two adult flukes were collected from naturally infected bovine livers in different areas from Argentina and used for DNA extraction. We carried out PCR amplification and sequence analysis of the ribosomal internal transcribed spacer 1 (ITS1), mitochondrial nicotinamide adenine dinucleotide dehydrogenase subunits 4 and 5 (nad4 and nad5) and mitochondrial cytochrome c oxidase subunit I (cox1) genes as genetic markers. Phylogenies were reconstructed using maximum parsimony algorithm. A total of 6 haplotypes were found for cox1, 4 haplotypes for nad4 and 3 haplotypes for nad5. The sequenced ITS1 fragment was identical in all samples. The analyzed cox1 gene fragment is the most variable marker and is recommended for future analyses. No geographic association was found in the Argentinean samples.


Simulation of responses of spectrally-opponent neurones in the macaque lateral geniculate nucleus to chromatic and achromatic light stimuli.
The responses of spectrally opponent, parvocellular-layer cells in the lateral geniculate nucleus of the macaque vary with stimulation along all three dimensions of colour space (luminance, dominant wavelength and purity). Here, the responses of different cell types to light stimuli successively replacing a white adaptation field are simulated mathematically by a simple model using known cone spectral sensitivities. Hyperbolic response functions with two adjustable parameters relate cone excitation with the output signals of each cone mechanism. For the majority of cells, differences of outputs of two cone mechanisms, receiving inputs from L- and M-cones, describe responses to chromatic and achromatic stimuli. For each opponent combination of cone inputs, either L-M or M-L, two cell types were found. Off-centre cells had lower maximum firing rates of their on-component, but were the more sensitive of the two groups, preferring dark colours, whereas on-centre cells showed better responsiveness to brighter stimuli. For all four cell types, the predictions of the equations replicated well the responses measured to stimuli varying along all three dimensions of colour space. The quantification of cell responses presented here provides possibilities for studying suprathreshold coding of colour and lightness, and the model can help to link physiology with the psychophysics of colour vision.


Methodological considerations on real-world evidence studies of monoclonal antibodies against the CGRP-pathway for migraine: a systematic review.
Real-world data are accumulating on the effectiveness, tolerability and safety of anti-calcitonin gene-related peptide pathway monoclonal antibodies for the preventive treatment of migraine. We performed a systematic review of the methodology of prospective, observational, clinic-based real-world evidence studies with these drugs in both episodic and chronic migraine. The objectives were to evaluate the definitions and reported outcomes used, and to perform a risk of bias assessment for each of the different studies. PubMed and EMBASE were systematically queried for relevant scientific articles. Study quality assessment of the included studies was conducted using the "National Heart, Lung and Blood Institute (NHLBI) Study Quality Assessment Tool for Before-After (Pre-Post) Studies with No Control Group". Forty-six studies fitted the criteria for the systematic review and were included in the analysis. Ten studies (21.7%) defined a migraine day for the study, while only 5 studies defined a headache day for the study (10.9%). The most common primary endpoint/objective of the studies was change in monthly migraine days (n = 16, 34.8%), followed by responder rate (n = 15, 32.6%) and change in monthly headache days (n = 5, 10.9%). Eight studies (17.4%) did not define the primary endpoint/objective. Thirty-three studies were graded as "good" quality and 13 studies were graded as "fair". Our analysis shows rather significant heterogeneity and/or lack of predefined primary outcomes/objectives, definitions of outcomes measures and the use of longitudinal monitoring (e.g. headache diaries). Standardization of terminology, definitions and protocol procedures for real-world evidence studies of preventive treatments for migraine are recommended. This study was registered with PROSPERO with ID CRD42022369366.


Palmitate turnover and its response to glucose infusion in weight-losing cancer patients.
Palmitate turnover in weight-stable control subjects (n = 4) and weight-losing patients with progressive malignant disease (n = 4) has been determined. Measurements were made after an overnight fast and during glucose infusion (3.5 mg/kg/min). Turnover rates were calculated from plateau isotopic enrichment of palmitate in plasma during a continuous infusion of 1-13C palmitate. Palmitate turnover was higher in the cancer group before (180%) and during glucose loading (170%) compared with the control group. Palmitate turnover was reduced during glucose administration by approximately 34% in both groups. Plasma concentration of insulin was decreased and of cortisol was increased in the cancer group compared with the control group before and during glucose infusion. We conclude that cancer patients with weight loss have increased rates of fatty acid turnover indicative of enhanced mobilisation of body fat stores. Altered plasma concentrations of insulin and cortisol may mediate this effect. Nonetheless, even at more advanced stages of cachexia cancer patients have normal control mechanisms for inhibiting fatty acid turnover following administration of carbohydrate.


Calculation of robustly relatively stabilizing PID controllers for linear time-invariant systems with unstructured uncertainty.
This article deals with the calculation of all robustly relatively stabilizing (or robustly stabilizing as a special case) Proportional-Integral-Derivative (PID) controllers for Linear Time-Invariant (LTI) systems with unstructured uncertainty. The presented method is based on plotting the envelope that corresponds to the trios of P-I-D parameters marginally complying with given robust stability or robust relative stability condition formulated by means of the H∞ norm. Thus, this approach enables obtaining the region of robustly stabilizing or robustly relatively stabilizing controllers in a P-I-D space. The applicability of the technique is demonstrated in the illustrative examples, in which the regions of robustly stabilizing and robustly relatively stabilizing PID controllers are obtained for a controlled plant model with unstructured multiplicative uncertainty and unstructured additive uncertainty. Moreover, the method is also verified on the real laboratory model of a hot-air tunnel, for which two representative controllers from the robust relative stability region are selected and implemented.


Environmental risk factors and Crohn's disease: a population-based, case-control study in Spain.
The pathogenesis of inflammatory bowel disease probably involves an interaction between genetic and environmental factors: cigarette smoking, appendectomy and oral contraceptives have been the factors most frequently linked to its aetiology To analyse the association between known environmental risk factors and development of Crohn's disease in the community of Aragón, Spain. A case-control, population-based study has been carried out. All patients diagnosed with Crohn's disease in the community of Aragón from 1st February 1992 to 31st January 1995 were prospectively included. The Lennard Jones criteria were used to define the cases and selected controls among the healthy population matched with patients for age, sex and rural/urban habitat. Statistical analysis included multivariate analysis using conditional logistic regression, testing 38 different models. A total of 103 patients were diagnosed with Crohn's disease in Aragón from 1st February, 1992 to 31st January, 1995. Of these 62 patients (60.2%) with Crohn's disease were smokers, compared with 42 (40.8%) controls (p<0.001). Cigarette smoking is considered a risk factor for Crohn's disease with an odds ratio of 3.09 (95% confidence interval, 1.58-6.05). After multivariate analysis, the positive association is maintained. A dose-dependent relation could not be demonstrated. No statistical differences (p=0.50) were detected in the analysis of previous appendectomy. Use of oral contraceptive acts as a risk factor with a p=0.048; odds ratio 2, 8, 95% confidence interval: 1.009-7.774; but this association disappears in the multivariate analysis. Eight patients had a family history (3 first degree and 5 second degree relatives) versus none of the controls (p=0.002). Of the variables studied for childhood hygiene none appeared significant. Smoking, family history, and oral contraceptive use, appear as risk factors for developing Crohn's disease in univariate analysis, but only smoking remains significant in all models of multivariate analysis.


["The blessed Leon Szancer Scholarships Foundation for Poor Students of the Jagiellonian Academy Faculty of Medicine" and its founder].
The paper is devoted to a small but significant episode in the history of the 19th century philanthropy. It describes one of numerous scholarship foundations existing then and a person of its founder, Leon Szancer (1802-1879). The Szancer's Foundation was created for students of medicine of the Jagiellonian University and 25 persons used its help. Szancer himself was a representative of Poles of Mosaic religion. He had graduated from the Jagiellonian University and afterwards he worked as physician in the Army of the Congress Kingdom of Poland. He participated in the Polish November Insurrection and after its fall he emigrated. He was working in Opatów for many years and spent his last years of life in Krakow.


Ramsay Hunt syndrome.
We report a case of a 26-year-old previously healthy male who presented with peripheral facial nerve palsy and painful vesicular eruptions over an erythematous base on pinna and external auditory canal. A diagnosis of Ramsay Hunt syndrome was made. Patient improved with prednisolone, acyclovir, and physiotherapy. Sometimes it may be difficult to differentiate Ramsay Hunt syndrome from Bell's palsy, as patient may develop vesiscles after the onset of facial weakness.


Association of Pro/Anti-inflammatory Cytokine Gene Polymorphisms with Benign Prostate Hyperplasia Risk.
Benign prostate hyperplasia (BPH) is a common condition in aged men and result from prolong chronic inflammation in prostate gland. Cytokines are important molecules responsible for inflammation. Single nucleotide polymorphisms (SNPs) in promoter region of cytokine genes have been shown to alter the level of cytokines. Hence we evaluated the association of pro-inflammatory and anti-inflammatory cytokine SNPs in a North Indian cohort of BPH patients. We observed that IL-1B -511 CT + TT genotypes conferred protective effect for susceptibility to BPH (OR 0.39, P 0.001). Our results also demonstrated that TNF-A -1031 C allele to be associated with risk for BPH (OR 1.89, P < 0.0001). Moreover, we also observed twofold risk for IL-10 -1082 cytokine gene polymorphism (OR 1.96, P 0.048). No association was observed with risk of BPH for IFN-G +874, IL-1 RN VNTR, IL-6 -174, IL-10 -819 and TGF-B +28. Our findings of IL-1B -511, TNF-A -1031 and IL-10 -1082 suggested that these variants play important role in susceptibility to BPH. Future studies in large cohort of different ethnicity BPH groups are warranted to establish definite associations with other cytokine gene polymorphisms as well.


Laparoscopic versus open approach for aortobifemoral bypass for severe aorto-iliac occlusive disease--a multicentre randomised controlled trial.
To investigate differences between open and laparoscopic aortobifemoral bypass surgery for aorto-iliac occlusive disease on postoperative morbidity and mortality. A multicentre randomised controlled trial. Between January 2007 and November 2009, 28 patients with severe aorto-iliac occlusive disease (TASC II C or D) were randomised between laparoscopic and open approach at one community hospital and one university hospital (TASC = Trans-Atlantic Inter-Society Consensus on the Management of Peripheral Arterial Disease). The operation time was longer for the laparoscopic approach (mean 4 h 19 min (2 h 00 min to 6 h 20 min) vs. 3 h 30 min (1 h 42 min to 5 h 11 min); p = 0.101)). Nevertheless, postoperative recovery and in-hospital stay were significantly shorter after laparoscopic surgery. Also oral intake could be restarted earlier (mean 20 h 34 min (6 h 00 min to 26 h 55 min) vs. 43 h 43 min (19 h 40 min to 77 h 30 min); p = 0.00014)) as well as postoperative mobilisation (walking) (mean 46 h 15 min (16 h 07 min to 112 h 40 min) vs. mean 94 h 14 min (66 h 10 min to 127 h 23 min); p = 0.00016)). Length of hospitalisation was shorter (mean 5.5 days (2.5-15) vs. mean 13.0 days (7-45); p = 0.0095)). Visual pain scores and visual discomfort scores were both lower after laparoscopic surgery. Also return to normal daily activities was achieved earlier. There were no major complications in both groups. Laparoscopic aortobifemoral bypass surgery for aorto-iliac occlusive disease is a safe procedure with a significant decrease in postoperative morbidity and in-hospital stay and earlier recovery.


[Bartonella henselae vertebral osteomyelitis: report of a case].
Cat scratch disease (CSD) is caused by Bartonella henselae, with unknown prevalence and incidence in the Chilean paediatric population. Regional lymphadenopathy is the most common presentation, while atypical forms constitute a diagnostic challenge. To report a case of CSD with osteomyelitis and present guidelines regarding treatment. An eight year-old patient, with prolonged febrile illness, back pain and neck stiffness. Laboratory studies highlight positive IgG for Bartonella henselae. The abdominal ultrasound showed splenic micro-abscesses, and the MRI showing vertebral lesions suggestive of osteomyelitis. The diagnosis of atypical forms requires a high rate of suspicion, as in this case, in which the patient manifested the musculoskeletal symptoms simultaneously with the febrile syndrome, which led us to study possible complications of the disease. Current knowledge of the treatment of atypical or complicated CSD is derived from the observation of case studies, rather than randomized trials. It is suggested that antibiotic therapy is analysed individually, with the help of a specialist. The importance of high clinical suspicion are emphasised and discussed, as well presenting some treatment options based on the evidence from the current literature.


The hypo-osmotic swelling test: Is it a sperm vitality or a viability assay?
The first two editions of the World Health Organization laboratory manual described the determination of live spermatozoa by a dye exclusion method as a sperm "viability" test, whereas subsequent editions classified it as a "vitality" test, without providing an explanation for the reclassification. Additionally, the hypo-osmotic swelling (HOS) test, which assesses the functional integrity of the human sperm membrane, was placed in the same category as the dye exclusion test. Although the two terms might seem synonymous, the term "vitality" merely means "alive," whereas "viability" assesses qualities or physiological functions of a living entity. After comparing the morphological, physiological, and clinical findings obtained from dye exclusion testing vs. the HOS test, we conclude that the HOS test should be classified as a viability test, not merely as a vitality test.


Haploidentical Peripheral Blood Stem Cell Transplantation with Post-Transplantation Cyclophosphamide in Children with Advanced Acute Leukemia with Fludarabine-, Busulfan-, and Melphalan-Based Conditioning.
Post-transplantation cyclophosphamide (PTCY) therapy has made haploidentical transplantation a global reality in adults, but the literature is largely silent on the feasibility of this approach in children. We conducted a prospective study of 20 patients (median age, 12 years; range, 2-20 years) with advanced acute leukemia to evaluate the feasibility of PTCY-based haploidentical peripheral blood stem cell (PBSC) transplantation in children. The conditioning regimen comprised fludarabine, i.v. busulfan, and melphalan (Flu-Bu-Mel). PTCY on days +3 and +4 was followed by mycophenolate mofetil for 14-21 days and cyclosporine for 60 days. Thirteen patients (65%) had refractory or relapsed myelogenous leukemia, and the remainder had high-risk lymphoblastic leukemia. Prompt engraftment was noted at a median of 14 days, with full donor chimerism by day +28. The cumulative incidence of acute and chronic graft-versus-host disease was 35% and 5%, respectively. Nonrelapse mortality at 1 year was 20%. The incidence of disease progression was 25.7%. The actuarial overall survival at 2 years was 64.3% (95% confidence interval, 53.4%-75.2%). Our data suggest that Flu-Bu-Mel-based conditioning followed by PTCY-based haploidentical PBSC transplantation with reduced duration of immunosuppression is feasible in pediatric patients with advanced leukemia.


Cyclic AMP selectively up-regulates calmodulin genes I and II in PC12 cells.
Calmodulin is encoded by three genes in rat tissues, which collectively transcribe five different mRNAs. We showed previously that the pheochromocytoma cell line, PC12, expresses all five of these mRNAs and that treating PC12 cells with nerve growth factor differentially induces the calmodulin mRNAs; the greatest increase was seen by 24 h in the 1.4 kb transcript from calmodulin gene II. In the present study we found that treating PC12 cells with dibutyryl cyclic AMP also differentially increased the levels of the calmodulin mRNAs. However, dibutyryl cyclic AMP produced increases as early as 3 to 6 h, with the greatest increase (about 3-fold) being seen in the level of the 1.7 kb mRNA transcribed from calmodulin gene I. The transcripts of 4.1 kb (calmodulin gene I) and 1.4 kb (calmodulin gene II) were also increased, but the 2.3 kb transcript from calmodulin gene III remained stable. Another cyclic AMP analogue, chlorophenylthio cyclic AMP, produced effects similar to those of dibutyryl cyclic AMP, but dibutyryl cyclic GMP did not. Pretreatment with cycloheximide blocked the increase in the 4.1 kb calmodulin mRNA induced by dibutyryl cyclic AMP, but only partially blocked the increase in the 1.4 kb and 1.7 kb transcripts. Phorbol 12-myristate 13-acetate, which can induce some cyclic AMP responsive genes, failed to significantly change the levels of any of the calmodulin mRNAs. These studies show that, like nerve growth factor, cyclic AMP can selectively up-regulate the transcripts from calmodulin genes in PC12 cells, but the time-course and type of transcripts induced by cyclic AMP are distinct. These results suggest that the mechanisms by which these two agents alter the calmodulin transcripts are different.


[Medical development cooperation in the Confederation. The international framework and the important place of public health in Swiss development cooperation].
The systematic organization of health services in the developing countries to benefit the majority of the population is seen as an important contribution toward the general economic and social development of these countries. This view is uncontested today, but was given less importance in the early years of Swiss development aid. The Swiss Development Cooperation's support of health services is now integrated into its overall program to satisfy the basic needs of the people of the developing countries.


A discriminant function analysis of various interferon parameters among alcoholics and heavy smokers.
A three-group design of alcoholics, heavy smokers and controls was used to investigate the status of the interferon system, including natural killer cell activity. Group differences were indicated via discriminant function analysis which correctly classified 86% of subjects. Test-retest relations were investigated for 14 subjects following a 30-day alcoholic inpatient program. Whereas significant immune suppression was indicated at the time of detoxification, recovery was evident at the 30-day follow-up. Results are discussed in terms of the significance of alcohol abuse on immune system functioning with consequences for susceptibility to viruses, bacteria and medical illnesses.


Evaluation of the therapeutic effect of melittin peptide on the ulcerative colitis mouse model.
Ulcerative colitis (UC) is considered one of the most prevalent inflammatory bowel diseases (IBDs). However, due to the lack of satisfying efficacy of conventional therapies and their side effects, there is still a need for more efficient therapeutic agents. Melittin is a small peptide derived from bee venom, which shows potent anti-inflammatory activity. The present investigation aimed to assess the anti-inflammatory effect of melittin peptide alone and in co-therapy with sulfasalazine as a standard therapy on dextran sulfate sodium (DSS)-induced colitis models. We used DSS to induce UC in C57BL/6 male mice. We investigated the effect of melittin peptide alone and in combination with sulfasalazine on improving the clinical symptoms among DSS-induced colitis models. Finally, we employed histological investigation to show the therapeutic effect of melittin on attenuating the pathological damage of colon tissue caused due to DSS-induced inflammation in colitis models. Our findings demonstrated that melittin peptide alone and in combination with sulfasalazine dramatically cured the clinical UC. Moreover, we observed that this peptide almost eliminated the histological damage of colon tissue in colitis, while significantly reducing the inflammation in colon tissue. Meanwhile, our results demonstrated that this peptide had an antioxidant effect through the disruption of the oxidant/antioxidant balance. All these findings suggest that melittin peptide has an anti-inflammatory effect and can probably be considered a novel therapeutic agent for UC. Furthermore, our results demonstrated that this peptide can enhance the therapeutic effects of conventional therapy while attenuating the adverse effects of conventional agents.


Evidence that 3-hydroxyisobutyric acid inhibits key enzymes of energy metabolism in cerebral cortex of young rats.
3-Hydroxyisobutyric aciduria is an inherited metabolic disease caused by 3-hydroxyisobutyryl-CoA dehydrogenase deficiency. Tissue accumulation and high urinary excretion of 3-hydroxyisobutyric acid is the biochemical hallmark of this disorder. Clinical phenotype is heterogeneous and generally includes dysmorphic features, delayed motor development, profound mental impairment, and acute encephalopathy. Lactic acidemia is also found in the affected patients, indicating that mitochondrial dysfunction may be involved in the pathophysiology of this disorder. Therefore, the aim of the present work was to investigate the in vitro effect of 3-hydroxyisobutyric acid (0.1, 0.5 and 1mM) on essential enzymes of energy metabolism, namely the activities of the respiratory chain complexes I-V, total, cytosolic and mitochondrial creatine kinase and Na(+), K(+)-ATPase in cerebral cortex homogenates of 30-day-old rats. We also measured the rate of oxygen consumption in brain mitochondrial preparations in the presence of 3-hydroxyisobutyric acid. 3-Hydroxyisobutyric acid significantly reduced complex I-III (20%), without affecting the other activities of the electron transport chain. Furthermore, 3-hydroxyisobutyric acid did not change state III, state IV and the respiratory control ratio in the presence of glutamate/malate or succinate, suggesting that its effect on cellular respiration was weak. On the other hand, the activities of total and mitochondrial creatine kinase, but not cytosolic creatine kinase, were inhibited (30%) by 3-hydroxyisobutyric acid. We also observed that 3-hydroxyisobutyric acid-induced inhibition of mitochondrial creatine kinase activity was fully prevented by pre-incubation of the homogenates with reduced glutathione, alpha-tocopherol or the combination of superoxide dismutase plus catalase, suggesting that this inhibition was mediated by oxidation of essential thiol groups of the enzyme probably by superoxide, hydrogen peroxide and/or peroxyl radicals. It was also demonstrated that Na(+), K(+)-ATPase activity from synaptic plasma membranes was markedly suppressed (37%) by 3-hydroxyisobutyric acid and that this effect was prevented by alpha-tocopherol co-incubation implying that peroxyl radicals were probably involved in this action. Considering the importance of the affected enzyme activities for brain metabolism homeostasis and neurotransmision, it is suggested that increased tissue levels of 3-hydroxyisobutyric acid may contribute to the neurodegeneration of patients affected by 3-hydroxyisobutyric aciduria and possibly explain previous reports describing elevated production and excretion of lactate.


High-Dose Methylprednisolone to Prevent Platelet Decline in Preeclampsia: A Randomized Controlled Trial.
To evaluate whether early administration of high-dose methylprednisolone limits the fall of platelets in preeclampsia. A randomized trial of 180 mg methylprednisolone or placebo administered in divided doses over 36 hours was conducted in women admitted for preeclampsia and platelet counts below 150×10/L in four French academic centers. Patients were not included when platelet counts were below 50×10/L or when immediate delivery was required. The primary study outcome was the proportion of patients with platelet counts above 100×10/L 36 hours after the first dose of study medication. The total sample size needed to detect a 23% difference in the rate of this outcome between groups with a one-tailed α of 0.05 and 90% power was 94 patients. Thirty-six patients were randomly assigned to receive methylprednisolone and 34 placebo between October 2007 and May 2011. Platelet counts above 100×10/L at 36 hours after the first dose of study medication were recorded in 30 (83%) in the active group and 29 (85%) in the placebo group (relative risk 0.98, 95% confidence interval 0.80-1.20; P=.82). The only adverse potentially study-related event was hyperglycemia in one woman allocated to methylprednisolone. In women with preeclampsia and platelet counts under 150×10/L, methylprednisolone was not effective in maintaining platelet counts above 100×10/L. EU Clinical Trials Register, http://clinicaltrialsregister.eu, EudraCT 2006-004881-15-FR.


Monoclonal anti-H antibody A46-B/B10 suited for differentiating blood groups A1 and A2 reveals also heterogeneity within blood group B.
A monoclonal antibody (mAb) recognizing H antigen was obtained from a BALB/c mouse immunized with MCF-7 (human mammary carcinoma) cells. This antibody (A46-B/B10, IgM) has been tested on a total of 753 blood samples. It strongly agglutinates human erythrocytes of blood groups 0, A2 and A2B, but not or only very weakly A1 and A1B. This mAb promises to be useful in the determination of A and AB subgroups in blood group serology. Evidence is presented that this antibody might also be able to define B subgroups. Among 263 B blood samples, 45 (17.1%) were negative or only weakly positive with A46-B/B10.


Fewer metabolites of dietary choline reach the blood of rats after treatment with lithium.
Choline is an important precursor for the biosynthesis of acetylcholine, phosphatidylcholine and sphingomyelin. It is also a major source of labile methyl groups. Lithium is an important component of the treatment of bipolar affective illness, and it inhibits choline transport across membranes. We studied the effect of lithium treatment upon the appearance in blood, liver and intestine of metabolites formed from dietary choline. Rats were treated for 9 days with 2 mEq/kg lithium carbonate or water. Animals were fasted overnight, and on the 10th day were fed with a solution containing radiolabeled choline chloride. The lithium-treated groups also received 2.0 mEq/kg lithium as part of this solution. After an oral dose of 1 ml of a 1 mM choline solution, the lithium-treated animals had significantly lower levels of choline-derived radiolabel in blood than did controls at 30, 60, 120, and 180 minutes (47% (+/- 5%; SEM), 51% (+/- 7%), 59% (+/- 4%) and 74% (+/- 9%), respectively). We observed similar decreases of the accumulation in blood, at 180 minutes after the dose, of choline-derived radiolabel when choline was administered at lower or higher concentrations. After an oral treatment containing 0.1, 1 or 10 mM choline, lithium treated animals accumulated 69% (+/- 6%; SEM), 66% (+/- 11%) and 72% (+/- 7%) as much radiolabel in serum as did controls. Most of the radiolabel found in blood at 180 minutes was in metabolites of choline which are formed within liver (betaine and phosphatidylcholine). The diminished accumulation of radiolabel in serum after lithium treatment was not due to increased accumulation of label by erythrocytes, liver or gut wall. We suggest that lithium influences the release by liver of betaine and phosphatidylcholine.


Nanobody-based chimeric receptor gene integration in Jurkat cells mediated by φC31 integrase.
The crucial role of T lymphocytes in anti-tumor immunity has led to the development of novel strategies that can target and activate T cells against tumor cells. Recombinant DNA technology has been used to generate non-MHC-restricted chimeric antigen receptors (CARs). Here, we constructed a panel of recombinant CAR that harbors the anti-MUC1 nanobody and the signaling and co-signaling moieties (CD3ζ/CD28) with different spacer regions derived from human IgG3 with one or two repeats of the hinge sequence or the hinge region of FcγRII. The PhiC31 integrase system was employed to investigate if the recombination efficiency could be recruited for high and stable expression of T cell chimeric receptor genes. The effect of nuclear localization signal (NLS) and two different promoters (CMV and CAG) on efficacy of PhiC31 integrase in human T cell lines was evaluated. The presence of integrase in combination with NLS, mediated up to 7.6 and 8.5 fold increases in CAR expression in ZCHN-attB and ZCHHN-attB cassette integrated T cells, respectively. Our results showed that highly efficient and stable transduction of the Jurkat cell line by PhiC31 integrase is a feasible modality for generating anti-cancer chimeric T cells for use in cancer immunotherapy.


Long-range chromosomal engineering is more efficient in vitro than in vivo.
Cre/LoxP mediated chromosomal engineering in embryonic stem (ES) cells has a variety of applications, including the creation of model systems for studying aneuploidy. Targeted meiotic recombination (TAMERE) was proposed as a high efficiency in vivo alternative to effect Cre-mediated recombination, in which Cre recombinase under control of the Synaptonemal Complex 1 promoter is expressed during male meiosis in transgenic mice. TAMERE has been successfully used with LoxP sites up to 100 kb apart. We tested TAMERE for a chromosome engineering application in which LoxP sequences were integrated into sites 3.9 Mb apart on the same (cis) or opposite (trans) copies of mouse Chromosome 16 (MMU16). TAMERE was ineffective in generating either a deletion or a translocation in vivo. The TAMERE method may be of limited use for large genomic rearrangements. The desired translocation was achieved with an in vitro method that can be used in any ES cell line. Mice produced from the reciprocal duplication/deletion of MMU16 in a region homologous to human chromosome 21 provide models that are useful in studies of Down syndrome.


Fastigial nucleus projections to the brain stem in beagles: pathways for autonomic regulation.
Efferent connections from a portion of the cerebellar fastigial nucleus were investigated using autoradiography. Bipolar stimulating electrodes were placed in the fastigial nucleus of anesthetized beagles and the area that produced increases in blood pressure and heart rate was localized. A mixture of [3H]leucine and [3H]proline (4:1) was injected into the area and autoradiograms of transported material were prepared. Injections filled the rostral and various parts of the caudal fastigial nucleus. Labeled axons reached the brain stem via two routes, the ipsilateral juxtarestiform body and the contralateral uncinate fasciculus. Ventral portions of the lateral vestibular nucleus were labeled bilaterally, projections to the inferior vestibular and medial vestibular nuclei are contralateral. Nucleus tractus solitarius was heavily labeled on the side opposite the injection. The contralateral medial reticular formation contained many labeled terminals and axons. Label was found in the nucleus reticularis ventralis, lateral reticular nucleus, nucleus gigantocellularis, nucleus pontis caudalis and the paramedian reticular nucleus. No terminal labeling was found in nucleus parvocellularis or nucleus ambiguous. Stimulation of the rostral fastigial nucleus produces increases in blood pressure and heart rate by generalized sympathoexcitation. Many cell groups which facilitate the activity of preganglionic sympathetic neurons do not receive direct fastigial input. It is suggested that that sympathoexcitation resulting from stimulation of the fastigial nucleus occurs through multisynaptic connections in the brain stem.


Dynamic generation of multi-qubit entanglement in the ultrastrong-coupling regime.
We propose a dynamic evolution protocol for generating multi-qubit GHZ states in the ultrastrong-coupling regime of circuit QED. By varying the time length of sequences, the protocol works for any coupling strength g/ωr ≥ 0.25. The time for generating the GHZ states in our protocol can be in the subnanoseconds. By taking into account realistic parameters of circuit QED, the degeneracy of fidelity due to decoherence can be as low as 0.02%.


Use of hepatitis C virus-positive grafts in liver transplantation: a single-centre experience.
Our goal was to evaluate the outcome of HCV(+) recipients after liver transplantation (LT) using HCV(+) donors and the interaction between donor and recipient viral strain. We performed a retrospective analysis of 21 LT performed between 1998 and 2004 using livers from HCV(+) donors in HCV(+) recipients. Two hundred thirty-seven patients with HCV cirrhosis who underwent LT with livers from HCV(-) donors were the control group. Ishak score (IS) was evaluated for all HCV(+) grafts. The considered variables included donor age, hepatic enzymes, intensive care unit stay, HCV genotype, ischemia time, recipient age, UNOS status, Child score, HCV genotype (before and 6 months after LT) and IS (after LT). We analyzed patient, graft, and disease-free survival. HCV(+) donors were significantly older than HCV(-) donors. The cumulative 5-year patient and graft survivals and disease free intervals were not different between groups. IS grading was more than 2/18 in two cases; the only graft with a staging score over 2/6 was retransplanted for early nonfunction. In two cases, different HCV genotypes were matched and donor strain took over the recipient strain. In one patient, donor genotyping 2a-2c took over recipient genotyping 1b and 9 months after LT recurrent hepatitis was documented, but antiviral therapy cleared HCV. Livers from HCV(+) donors can safely be used in HCV(+) recipients. Hepatic biopsy must always be performed; livers with bridging fibrosis should not be used. The takeover of one strain by another may change the prognosis of the patient if the predominant strain is more sensitive to antiviral therapy.


[Confidential healthcare for Israeli adolescents: towards evolution of specific legislation].
Despite its critical importance, the issue of confidentiality in the provision of adolescent health care has not been specifically clarified by Israeli law. To review the conflict between the needs of adolescents, attitudes of the medical community and daily medical practice, and to promote legislation that would allow health care providers to treat adolescents without parental consent under certain circumstances. A major key to the productive encounter between the adolescent and the health professional is the confidence that sensitive information, communicated between them, will not be divulged to anyone, including parents. The Israeli law does not allow minors to give independent consent to care, except under certain circumstances. Also, Israeli physicians frequently do not provide confidential care, even when the issue is explicitly covered by the law such as in the case of termination of pregnancy. The Ministry of Health adopted an analogy stating that "in the case of an activity in which minors of his/her age are accustomed to engaging, a legal act performed by a minor cannot be cancelled". On this basis, medical associations and other bodies involved in adolescent health recommended that care related to issues requiring privacy should be provided to minors without parental consent or knowledge whenever it is common among a certain age group to seek the specific type of care. However, a clear and updated legislation is still required in order to enhance professional legitimization and awareness.


Effect of changes in body mass index and waist-to-height ratio on blood pressure in 11- to 13-year-old children: a prospective population study.
Existing evidence about associations between change in body mass index (BMI) and waist-to-height ratio (WHtR) change and high blood pressure are relatively limited. We aimed to investigate the associations of general overweight (based on BMI) and abdominal obesity (based on WHtR) change with high blood pressure in Chinese children. A school-based cohort study in Ningbo region (China) was conducted among children with baseline evaluations in October 2016 with follow-up two years later. A total of 1432 children aged 11-13 years participated in this study. Our results showed that a change from normal BMI or WHtR to overweight or abdominal obesity in children was associated with high blood pressure (adjusted odds ratio (AOR), 2.62; p<0.05 or AOR, 2.79; p<0.05, respectively). In addition, an increased risk of high blood pressure was observed in children who maintained overweight or abdominal obesity (AOR, 1.67; p<0.05 or AOR, 1.69; p<0.05, respectively), but not in children who experienced remission to non-excess weight. Interestingly, children who increased BMI or WHtR had greater impact on SBP than on DBP. The 2-year longitudinal study indicated that general overweight or abdominal obesity can predict the risk factor of high blood pressure in children. However, children who remitted to non-excess weight did not exhibit an increased risk of high blood pressure.


A time course investigation of vitamin A level and lipid composition of the liver endoplasmic reticulum in rats following treatment with congeneric polychlorobiphenyls.
The drug metabolizing enzyme activities, the vitamin A content and the fatty acid composition in the endoplasmic reticulum membrane were studied in rat liver after a single injection of the polychlorobiphenyls (PCBs) 3,3',4,4'-tetrachlorobiphenyl [(3,4)2Cl] or 2,2',4,4',5,5'-hexachloro-biphenyl [(2,4,5)2Cl], 300 mumol/kg each. The microsomal vitamin A level was markedly lowered 3 days after treatment with (3,4)2Cl, a coplanar type inducer of cytochrome P-450. A marked increase in microsomal AHH and UDPGT activities occurred within 3 days after injection of (3,4)2Cl whereas (2,4,5,)2Cl treatment enhanced APDM activity only. Arachidonic, stearic and linoleic acid microsomal contents were enhanced by the two congeners. (3,4)2Cl caused the proportion of docosahexaenoic acid to decrease. No highly significant correlation was found between the vitamin A content and lipid components in the microsomal membrane. However, the vitamin A level was inversely related to the activities of drug metabolizing enzymes induced by coplanar compounds (cytochrome P-450 towards benzo[a]pyrene and UDP glucuronosyl transferase towards 4-nitrophenol).


Diagnostic radiology in disaster medicine: implications for design, planning and organization of X-ray departments.
Diagnostic radiology plays an important role in the evaluation of disaster casualties; these are referred to X-ray departments, shortly after their arrival at hospital, in large numbers, frequently overwhelming facilities and resources of the department. The study of relevant literature and the experience of the authors suggest that there are implications for design, planning and organization of X-ray departments to be considered in disaster-prone areas. Departments should be sited as near as possible to triage areas, on the same floor, with broad, unobstructed passages. There should be provision for easy passage of equipment and trolleys through doors and corridors and large circulation areas, working rooms and waiting spaces with plugs, for electricity, piped oxygen and suction and devices for hanging infusion sets. Two entrances to the department would enable one-way flow through the department. All work should be done in the department and examinations performed with mobile units avoided. A radiologist at the entrance to the department and a control post at the exit ensure adequate processing of casualties. Interpretation and reporting should be done by assigned radiologists dictating directly to typists; films and reports should accompany casualties.


Interfacial activity of metal ion extractant.
The interfacial activity of metal ion extractants is discussed. The interfacial tension isotherms are processed by matching the Szyszkowski equation, and estimation of selected parameters to discuss the interfacial activity of extractants. Results are presented in comprehensive tables and figures. The interfacial activity depends mainly on three different parameters; type of extractant, acidity of the aqueous phase and organic diluent. Strongly acidic extractants and protonated amine reagents exhibit the highest interfacial activity. The solvating reagents and non-protonated amines are on the opposite side of the scale, while the chelating reagents are somewhere in the middle. The acidity of the aqueous phase affects the interfacial activity of extractants, mainly by the ionisation (protonation and dissociation) of extractant molecules. Solvating diluents always decrease the interfacial activity of extractants.


Effects of muscular and mental fatigue on spatiotemporal gait parameters in dual task walking in young, non-frail and frail older adults.
Dual-task (DT) walking is of great interest in clinical evaluation to evaluate frailty or cognitive declines in older adults. Frail older adults are known to adopt different walking strategy to overcome fatigue. However, no studies evaluated the effect of muscular or mental fatigue on dual-task walking strategy and the difference between frail and non-frail older adults. Evaluate the effect of mental and muscular fatigue on spatio-temporal parameters in dual-task walking in young, non-frail and frail older adults. 59 participants divided into 20 young (Y) (24.9 ± 3 years old), 20 non-frail (NF) (75.8 ± 4.9 years old) and 19 frail older adults (F) (81 ± 4.7 years old) performed single-task (ST) walking, single-task cognitive (serial subtraction of 3), and dual-task (subtraction + walking) for 1 min at their fast pace. Gait speed, step length, step length variability, stance and swing phase time, single and double support time, cadence, gait speed variability were recorded in single- and dual-task walking. The dual-task effect (DTE) was calculated as ((DT - ST)/ST) × 100). Generalized linear mixed models (GLMM) were used to compare the effects of mental and muscular fatigue on gait and cognitive variables between the groups. The DTE walking parameters were worse in F compared to NF or Y but no significant effect of fatigue were highlighted except for swing time and single support time DTEs. The results were mitigated but a clear difference in dual-task spatio-temporal parameters was found between F and NF which brings hope into the capacity of DT to better reveal frailty.


Naming and categorizing objects: task differences modulate the polarity of semantic effects in the picture-word interference paradigm.
The picture-word interference paradigm is a prominent tool for studying lexical retrieval during speech production. When participants name the pictures, interference from semantically related distractor words has regularly been shown. By contrast, when participants categorize the pictures, facilitation from semantically related distractors has typically been found. In the extant studies, however, differences in the task instructions (naming vs. categorizing) were confounded with the response level: While responses in naming were typically located at the basic level (e.g., "dog"), responses were located at the superordinate level in categorization (e.g., "animal"). The present study avoided this confound by having participants respond at the basic level in both naming and categorization, using the same pictures, distractors, and verbal responses. Our findings confirm the polarity reversal of the semantic effects--that is, semantic interference in naming, and semantic facilitation in categorization. These findings show that the polarity reversal of the semantic effect is indeed due to the different tasks and is not an artifact of the different response levels used in previous studies. Implications for current models of language production are discussed.


Expression of nitrotyrosine is associated with angiogenesis in esophageal squamous cell carcinoma.
In our previous study, it was suggested that nitrotyrosine, a product of nitrogen species, found in esophageal squamous cell carcinoma (ESCC), may contribute to the progression of esophageal cancer. To clarify whether nitrotyrosine expression is associated with apoptosis and/or angiogenic factors in ESCC, we have analyzed the relationship between nitrotyrosine presence and the apoptosis-related proteins, Bcl-2 and Bax, or CD34, the marker of vascular endothelial cells, by an immunohistochemical approach. Nitrotyrosine was detected in 21 out of 55 esophageal cancers. The correlation between nitrotyrosine presence and Bcl-2, Bax expression or apoptotic index (AI) was not significant. In contrast, nitrotyrosine presence was significantly correlated with the microvessel density (MVD); nitrotyrosine-positive specimens tended to show a high MAD, while nitrotyrosine-negative specimens tended to be associated with a low MVD (p<0.05). Our data suggest that NO induces progression of esophageal carcinoma through its effect on angiogenesis, rather than its effect on tumor apoptosis.


Persistence of left ventricular hypertrophy is associated with increased cardiovascular morbidity and mortality in hypertensive patients with lower achieved systolic pressure during antihypertensive treatment.
To determine if persistence of electrocardiographic (ECG) left ventricular hypertrophy (LVH) during aggressive systolic blood pressure (SBP) lowering would identify patients at increased risk. Adjudicated outcomes were examined in relation to the presence of LVH by mean in-treatment Cornell product (CP) in 463 hypertensive patients with mean in-treatment SBP ≤ 130 mmHg randomly assigned to losartan- or atenolol-based treatment. During mean follow-up of 4.4 ± 1.3 years, persistence of mean CP > 2440 mm ms in 211 patients (45.6%) was associated with significantly higher 4-year rates of cardiovascular death (8.9% vs 3.4%, p = 0.003), myocardial infarction (7.0% vs 3.3%, p = 0.010), stroke (8.5% vs 2.1%, p = 0.002), the composite endpoint of these events (20.0% vs 7.0%, p < 0.001) and all-cause mortality (14.9% vs 10.0%, p = 0.015). In multivariate Cox analyses, adjusting for a propensity score for CP LVH, randomized treatment and Framingham risk score entered as standard covariates and in-treatment diastolic BP and Sokolow-Lyon voltage LVH entered as time-varying covariates, persistence of CP LVH remained associated with statistically significant increased risks of cardiovascular death (hazard ratio, HR = 2.51, 95% CI 1.10-5.70), stroke (HR = 2.63, 95% CI 1.03-6.97) and the composite endpoint (HR = 2.46, 95% CI 1.36-4.45). These findings suggest that persistence of LVH in a subset of these patients may in part explain the lack of benefit found in hypertensive patients despite treatment to lower SBP.


Atypical sleep in critically ill patients on mechanical ventilation is associated with increased mortality.
Sleep patterns in critically ill patients' polysomnographic sleep studies (PSG) are severely abnormal. We aimed to investigate the association of atypical sleep patterns, micro-sleep phenomena (sleep spindles and K-complexes) and rapid eye movement (REM) sleep with intensive care unit (ICU), in-hospital and 90-day mortality in conscious critically ill patients on mechanical ventilation. This was a prospective descriptive study. We analysed 52 PSGs recorded in conscious critically ill patients on mechanical ventilation. PSGs were scored according to standard classification when possible. Otherwise, modified classification proposed for scoring sleep in critically ill patients was used. The association of PSG findings with mortality was studied using logistic regression and Weibull model of survival analysis. The presence of atypical sleep patterns in accordance with modified sleep scoring classification was associated with higher odds for ICU mortality (odds ratio 11.63; p = 0.03). The absence of K-complexes was associated with higher odds for ICU mortality (odds ratio 11.63; p = 0.03), while the absence of sleep spindles was associated with higher odds for in-hospital (odds ratio 7.80; p = 0.02) and 90-day mortality (odds ratio 5.51; p = 0.02). Loss of sleep spindles was associated with higher mortality risk with cutoff point 90 days (hazard ratio 3.87; p = 0.03). The presence of atypical sleep and absence of normal PSG sleep characteristics in conscious critically ill patients on mechanical ventilation indicates involvement of sleep producing brain structures in the pathological process and is associated with poor outcome.


Cannabinoid receptor-2 immunoreactivity is associated with survival in squamous cell carcinoma of the head and neck.
The prediction of progression of individual tumours, prognosis, and survival in squamous cell carcinoma (SCC) of the head and neck is difficult. Cannabinoid-1 (CB1) and cannabinoid-2 (CB2) receptor expression is related to survival in several types of cancer, and the aim of this study was to find out whether the expression of CB1 and CB2 receptors is associated with survival in primary SCC of the head and neck. We made immunohistochemical analyses of the cannabinoid receptors on tissue arrays from 240 patients with the disease. Receptor immunoreactivity was classified as none, weak, moderate, or strong staining. Overall survival and disease-specific survival were plotted using Kaplan-Meier survival curves. A multivariate Cox proportional hazard model was created with all the relevant clinical and pathological features. Strong immunoreactivity of the CB2 receptor was significantly associated with reduced disease-specific survival (p=0.007). Cox-proportional hazard ratio (HR) showed that CB2 receptor immunoreactivity contributed to the prediction of survival (HR 3.6, 95% CI 1.5-8.7, p=0.004). Depth of invasion (HR 2.2, 95% CI 1.2-4.2, p=0.01) and vascular invasion (HR 2.5, 95% CI 1.4-4.5, p=0.001) were also associated with survival.


Urological management in the patient with acute spinal cord injury.
Understanding the neurophysiology of micturition has led to pharmacologic manipulation and improvement in rates of mortality and morbidity. Use of intermittent self-catheterization and the artificial urinary sphincter (AUS) has improved the "quality of life" of these patients. Implantation of various penile prostheses has improved sexual rehabilitation. Urinary diversion is the "last resort" surgery.


Microarray analysis of the differential transformation mediated by Kirsten and Harvey Ras oncogenes in a human colorectal adenocarcinoma cell line.
Colorectal cancer arises after a series of mutational events in the colon epithelia and is often used as a model of the multistep progression of tumorigenesis. Mutations in Ki-Ras have been detected in some 50% of cases and are thought to occur at an early stage. Almost never do mutations arise in the loci of other Ras isoforms (Ha- and N-), leading to the assumption that Ki-Ras plays a unique role in tumorigenesis. In order to examine the distinctive function that Ki-Ras plays in cancer development in the colon, we introduced constitutively active mutant Ki- and Ha-Ras genes into an intermediate-stage colon adenoma cell line (Caco-2). We found that mutant active Ha-RasV12 was more efficient at transforming these colon epithelial cells as assessed by anchorage-independent growth, tumor formation in SCID mice and the development of mesenchymal morphology compared to transformation by Ki-RasV12. We conducted microarray analysis in an attempt to reveal the genes whose aberrant expression is a direct result of overexpression of either Ki-RasV12 or Ha-RasV12. We used Clontech's Atlas cancer cDNA (588 genes) and RZPD's Onco Set 1 (1,544 genes) arrays. We identified fewer genes that were commonly regulated than were differentially expressed between Ki- and Ha-RasV12 isoforms. Specifically, we found that Ki-RasV12 regulated genes involved in cytokine signaling, cell adhesion and colon development, whereas Ha-RasV12 mainly regulated genes involved in controlling cell morphology, correlating to an epithelial-mesenchymal transition only observed in these cells. Our results demonstrate how 2 Ras isoforms regulate disparate biologic processes, revealing a number of genes whose deregulated expression may influence colon carcinogenesis (supplementary material for this article can be found on the International Journal of Cancer website at http://www.interscience.wiley.com/jpages/0020-7136/suppmat/index.html).


An audit of patient acceptance of one-stop diagnosis for symptomatic breast disease.
The impetus for optimizing outpatient provision of breast-care services has come both from the patient and management in order to reduce anxiety and make full use of scarce resources. The one-stop diagnostic clinic for the investigation of symptomatic breast lesions is a relatively recent concept with well-known service benefits. However, acceptance to the patient has not been previously investigated. The results of this prospective audit demonstrate a high level of patient satisfaction with the multi-disciplinary, one-stop breast clinic.


A multi-analyte serum test for the detection of non-small cell lung cancer.
In this study, we appraised a wide assortment of biomarkers previously shown to have diagnostic or prognostic value for non-small cell lung cancer (NSCLC) with the intent of establishing a multi-analyte serum test capable of identifying patients with lung cancer. Circulating levels of 47 biomarkers were evaluated against patient cohorts consisting of 90 NSCLC and 43 non-cancer controls using commercial immunoassays. Multivariate statistical methods were used on all biomarkers achieving statistical relevance to define an optimised panel of diagnostic biomarkers for NSCLC. The resulting biomarkers were fashioned into a classification algorithm and validated against serum from a second patient cohort. A total of 14 analytes achieved statistical relevance upon evaluation. Multivariate statistical methods then identified a panel of six biomarkers (tumour necrosis factor-α, CYFRA 21-1, interleukin-1ra, matrix metalloproteinase-2, monocyte chemotactic protein-1 and sE-selectin) as being the most efficacious for diagnosing early stage NSCLC. When tested against a second patient cohort, the panel successfully classified 75 of 88 patients. Here, we report the development of a serum algorithm with high specificity for classifying patients with NSCLC against cohorts of various 'high-risk' individuals. A high rate of false positives was observed within the cohort in which patients had non-neoplastic lung nodules, possibly as a consequence of the inflammatory nature of these conditions.


Mutual Influences on Bullying Perpetration and Substance Use Among Adolescents in the United States.
Identifying temporal pattern of associations between bullying perpetration and substance use among youth is useful for optimal targeting of intervention and prevention. We examined the mutual influences on bullying perpetration and substance use and test the cumulative effects of bullying perpetration and substance use over time. We used data from the Population Assessment of Tobacco and Health cohort study, a nationally representative sample of youth, on bullying perpetration and substance use at three points in time over 3 years. We analyzed the associations of bullying perpetration with alcohol, cannabis and cigarette use using binary logistic regressions. The 13,068 youths aged 12-17 years at baseline (2013-2014) were included in the analytical sample. Cross-sectional associations between bullying perpetration and substance use were found at baseline (odds ratio [OR]alcohol = 1.62, 95% confidence interval [CI] 1.33-1.97; ORmarijuana = 1.28, 95% CI .99-1.66; ORcigarette = 1.86, 95% CI 1.44-2.39). Cumulative episodic and frequent bullying perpetration over three data waves predicted an increased likelihood of substance use at follow-up (ORs between 1.18, 95% CI 1.02-1.36 and 2.13, 95% CI 1.66-2.74). Conversely, both episodic and frequent substance use predicted an increased probability of bullying others at follow-up (ORs between 1.50, 95% CI 1.20-1.86, and 2.10, 95% CI 1.69-2.61). Associations between bullying perpetration and substance use appears to be bidirectional. In addition, episodic bullying perpetrators displayed intermediate levels of risk, suggesting a graded association between bullying perpetration pattern and substance use.


A Gene Expression Analysis of M1 and M2 Polarized Macrophages.
Macrophages are professional innate immune cells that are broadly disseminated throughout the body, shape various innate and adaptive immune responses, and play crucial roles in inflammation, homeostasis, wound healing, and tissue remodelling. According to their surrounding microenvironments, macrophages can differentiate themselves in different phenotypes. Over the last two decades, gene expression profiling has been used to decipher new transcripts associated with macrophage phenotypes. This chapter outlines protocols used to isolate and culture murine macrophages and how they can be "polarized" to obtain a specific phenotype. Furthermore, we describe a protocol for gene expression profiling using a quantitative real-time polymerase chain reaction (qPCR), a high-standard technology in the field of gene expression.


A highly immunogenic trivalent T cell receptor peptide vaccine for multiple sclerosis.
T cell receptor (TCR) peptide vaccination is a novel approach to treating multiple sclerosis (MS). The low immunogenicity of previous vaccines has hindered the development of TCR peptide vaccination for MS. To compare the immunogenicity of intramuscular injections of TCR BV5S2, BV6S5 and BV13S1 CDR2 peptides in incomplete Freunds adjuvant (IFA) with intradermal injections of the same peptides without IFA. MS subjects were randomized to receive TCR peptides/IFA, TCR peptides/saline or IFA alone. Subjects were on study for 24 weeks. The TCR peptides/IFA vaccine induced vigorous T cell responses in 100% of subjects completing the 24-week study (9/9) compared with only 20% (2/10) of those receiving the TCR peptides/saline vaccine (P =0.001). IFA alone induced a weak response in only one of five subjects. Aside from injection site reactions, there were no significant adverse events attributable to the treatment. The trivalent TCR peptide in IFA vaccine represents a significant improvement in immunogenicity over previous TCR peptide vaccines and warrants investigation of its ability to treat MS.


Tenofovir disoproxil fumarate.
Tenofovir disoproxil fumarate (tenofovir DF) is a prodrug of tenofovir, a nucleotide reverse transcriptase inhibitor. In two large, well designed, placebo-controlled clinical trials, tenofovir DF 300 mg/day resulted in significant reductions in HIV-1 RNA from baseline compared with placebo at 24 weeks in antiretroviral-experienced patients with HIV infection. Patients in both treatment groups continued to receive existing stable antiretroviral therapy. In an extension phase of one trial, these reductions in viral load were maintained after 96 weeks of treatment with tenofovir DF. Preliminary data from a large, 3-year comparative trial suggest the clinical efficacy of tenofovir DF in combination with baseline antiretroviral therapy is similar to that of stavudine in antiretroviral-naive patients with HIV infection. Virological substudies showed that viral suppression was maintained in patients who developed new reverse transcriptase mutations during tenofovir DF therapy (in combination with existing stable antiretroviral drugs) for up to 48 weeks. Isolates of HIV infrequently developed the K65R mutation during 96 weeks of tenofovir DF therapy. Tenofovir DF is generally well tolerated. The most commonly observed adverse events seen with tenofovir DF (in combination with other antiretroviral drugs) were predominantly of a gastrointestinal nature.