Immunoinformatics for Novel Multi-Epitope Vaccine Development in Canine Parvovirus Infections.
Canine parvovirus (CPV-2) is one of the most important pathogens of dogs of all ages, causing pandemic infections that are characterized by fatal hemorrhagic enteritis. The CPV-2 vaccine is recommended as a core vaccine for pet animals. Despite the intensive practice of active immunization, CPV-2 remains a global threat. In this study, a multi-epitope vaccine against CPV-2 was designed, targeting the highly conserved capsid protein (VP2) via in silico approaches. Several immunoinformatics methods, such as epitope screening, molecular docking, and simulation were used to design a potential vaccine construct. The partial protein sequences of the VP2 gene of CPV-2 and protein sequences retrieved from the NCBI were screened to predict highly antigenic proteins through antigenicity, trans-membrane-topology screening, an allergenicity assessment, and a toxicity analysis. Homologous VP2 protein sequences typically linked to the disease were identified using NCBI BLAST, in which four conserved regions were preferred. Overall, 10 epitopes, DPIGGKTGI, KEFDTDLKP, GTDPDDVQ, GGTNFGYIG, GTFYFDCKP, NRALGLPP, SGTPTN, LGLPPFLNSL, IGGKTG, and VPPVYPN, were selected from the conserved regions to design the vaccine construct. The molecular docking demonstrated the higher binding affinity of these epitopes with dog leukocyte antigen (DLA) molecules. The selected epitopes were linked with Salmonella enterica flagellin FliC adjuvants, along with the PADRE sequence, by GGS linkers to construct a vaccine candidate with 272 nucleotides. The codon adaptation and in silico cloning showed that the generated vaccine can be expressed by the E. coli strain, K12, and the sequence of the vaccine construct showed no similarities with dog protein. Our results suggest that the vaccine construct might be useful in preventing canine parvoviral enteritis (CPE) in dogs. Further in vitro and in vivo experiments are needed for the validation of the vaccine candidate.


Survival Benefit of Neoadjuvant Treatment in Clinical T3N0M0 Esophageal Cancer: Results From a Retrospective Multicenter European Study.
Based on current guidelines, clinical T3N0M0 esophageal tumors may or may not receive neoadjuvant treatment, according to their perception as locally advanced (cT3) or early-stage tumors (stage II). The study aim was to assess the impact of neoadjuvant treatment upon survival for cT3N0M0 esophageal cancer patients, with subgroup analyses by histological type (squamous cell carcinoma vs adenocarcinoma) and type of neoadjuvant treatment (chemotherapy vs radiochemotherapy). Data from patients operated on for esophageal cancer in 30 European centers were collected. Among the 382 of 2944 patients with clinical T3N0M0 stage at initial diagnosis (13.0%), we compared those treated with primary surgery (S, n = 193) versus with neoadjuvant treatment plus surgery (NS, n = 189). The S and NS groups were similar regarding their demographic and surgical characteristics. In-hospital postoperative morbidity and mortality rates were comparable between groups. Patients were found to be pN+ in 64.2% versus 42.9% in the S and NS groups respectively (P < 0.001), pN2/N3 in 35.2% versus 21.2% (P < 0.001), stage 0 in 0% versus 16.4% (P < 0.001), and R0 in 81.3% versus 89.4% of cases (P = 0.026). Median overall and disease-free survivals were significantly better in the NS group, 38.4 versus 27.9 months (P = 0.007) and 31.6 versus 27.5 months (P = 0.040), respectively, and this difference remained for both histological types. Radiotherapy did not offer a benefit compared with chemotherapy alone (P = 0.687). In multivariable analysis, neoadjuvant treatment was an independent favorable prognostic factor (HR 0.76, 95% CI 0.58-0.99, P = 0.044). Neoadjuvant treatment offers a significant survival benefit for clinical T3N0M0 esophageal cancer.


Brachial Plexus Injury due to Central-line Insertion during Kypho-scoliosis Deformity Correction.
Internal Jugular Venous (IJV) cannulation or central-line insertion is frequently performed during kyphoscoliosis deformity correction surgery or spine surgery with high risk. This helps monitor central venous pressure and administer medicines when required. Although many complications of IJV cannulation have been reported in the literature, its effect on brachial plexus is not known. The objective of this paper was to report a rare complication of IJV during scoliosis surgery. We reported a case of 27-year-old male who was operated for severe kyphoscoliosis correction where preoperatively IJV cannulation was done. Repeated attempts were done during IJV cannula insertion due to altered anatomy. Eventually, cannula insertion was done using ultrasound modality and surgery for correction was done. Postoperatively patient developed right upper extremity weakness and sensory loss although the clinical result of kyphoscoliosis correction was acceptable. EMG-NCV study proved it brachial plexus injury. The patient was treated with intravenous steroid and physiotherapy. The patient recovered completely within 6 months of surgery. We reported a case of kyphoscoliosis deformity corrective surgery where IJV cannulation led to brachial plexus injury and was eventually recovered with medications and physiotherapy.


Transforming growth factor-beta1 mediates coexpression of the integrin subunit alphaE and the chymase mouse mast cell protease-1 during the early differentiation of bone marrow-derived mucosal mast cell homologues.
Mucosal mast cells (MMC) play a central role in gut hypersensitivities and inflammation. They are morphologically, biochemically and functionally distinct from their connective tissue counterparts. Massive hyperplasia of MMC occurs 7-10 days after intestinal infection with nematodes but it has never been possible to replicate this phenomenon in vitro. (1) To determine whether mouse bone marrow-derived mast cells (mBMMC) grown in the presence of transforming growth factor (TGF)-beta1 could develop over the same time frame (7-10 days) as MMC in parasitized mice. (2) To compare the early expression of surface receptors (integrins alphaE and beta7, c-kit and FcepsilonR) with that of the MMC-specific granule chymase mouse mast cell protease-1 (mMCP-1). Mouse bone marrow cells were cultured in the presence of IL-9, IL-3 and Stem Cell Factor (SCF) with or without TGF-beta1. mBMMC were quantified after toluidine blue or Leishmans' staining. Expression of MMC-specific mouse mast cell proteases was analysed by ELISA, immunohistochemistry and RT-PCR. Surface antigen expression was characterized by flow cytometry and confocal microscopy. TGF-beta1 promotes the development of abundant MMC-like mBMMC from bone marrow progenitor cells with kinetics, which closely parallel that seen in vivo. mRNA transcripts encoding mMCP-1 and -2 are readily detectable by day 4 ex vivo in cultures grown in the presence of TGF-beta1. Between 30 and 40% and 75-90% of the cells in these cultures on days 4 and 7, respectively, have typical mast cell morphology, are c-kit+, FcepsilonR+, integrin alphaEbeta7+, and express and secrete abundant mMCP-1. The integrin alphaE subunit is coexpressed with mMCP-1. The kinetics of mMCP-1+/alphaE+ mBMMC development, regulated by TGF-beta1, are consistent with that seen in vivo in the parasitized intestine. The normally down-regulatory functions of TGF-beta1 in haematopoiesis are superseded in this culture system by its ability to promote the early expression of alphaE and mMCP-1.


[The mitochondrial membrane potential and oxygen consumption in the heart of spontaneously hypertensive rats].
We investigated the mitochondrial membrane potential and processes of respiration and oxidative phosphorylation in suspension of cardiac mitochondria from 6 month old spontaneously hypertensive rats (SHR) and Wistar (as a control) male rats. The mitochondrial membrane potential and the speed of oxygen consumption were measured using the method described by M.Brand (1995). Processes of respiration and oxidative phosphorylation in cardiac mitochondria were measured using Oxygraph (Hansatech instruments, Norfolk, England). It has been found that in SHR the mitochondrial membrane potential was lower (-113.76 mV+/-3.65 mV) compared to Wistar rats (DeltaPsim=-152.85 mV+/-13.52 mV, p<0.01). In SHR, the respiration rate in state V2 by Chance and V3 were increased compared to Wistar rats (p<0.001). The respiration control by Chance was depressed by 23.9% in SHR compared to Wistar rats. Our data demonstrate, that SHR have some features of functioning of cardiac mitochondria which distinguish them from the Wistar rats. Our data suggest a functional link between mitochondrial energy supply and arterial hypertension.


Activity of MM-398, nanoliposomal irinotecan (nal-IRI), in Ewing's family tumor xenografts is associated with high exposure of tumor to drug and high SLFN11 expression.
To determine the pharmacokinetics and the antitumor activity in pediatric cancer models of MM-398, a nanoliposomal irinotecan (nal-IRI). Mouse plasma and tissue pharmacokinetics of nal-IRI and the current clinical formulation of irinotecan were characterized. In vivo activity of irinotecan and nal-IRI was compared in xenograft models (3 each in nu/nu mice) of Ewing's sarcoma family of tumors (EFT), neuroblastoma (NB), and rhabdomyosarcoma (RMS). SLFN11 expression was assessed by Affymetrix HuEx arrays, Taqman RT-PCR, and immunoblotting. Plasma and tumor concentrations of irinotecan and SN-38 (active metabolite) were approximately 10-fold higher for nal-IRI than for irinotecan. Two doses of NAL-IRI (10 mg/kg/dose) achieved complete responses maintained for >100 days in 24 of 27 EFT-xenografted mice. Event-free survival for mice with RMS and NB was significantly shorter than for EFT. High SLFN11 expression has been reported to correlate with sensitivity to DNA damaging agents; median SLFN11 mRNA expression was >100-fold greater in both EFT cell lines and primary tumors compared with NB or RMS cell lines or primary tumors. Cytotoxicity of SN-38 inversely correlated with SLFN11 mRNA expression in 20 EFT cell lines. In pediatric solid tumor xenografts, nal-IRI demonstrated higher systemic and tumor exposures to SN-38 and improved antitumor activity compared with the current clinical formulation of irinotecan. Clinical studies of nal-IRI in pediatric solid tumors (especially EFT) and correlative studies to determine if SLFN11 expression can serve as a biomarker to predict nal-IRI clinical activity are warranted.


Molecular Iodine-Catalyzed Aerobic α,β-Diamination of Cyclohexanones with 2-Aminopyrimidine and 2-Aminopyridines.
Molecular iodine is shown to be an excellent catalyst for aerobic oxidative α,β-diamination of cyclohexanones with 2-aminopyrimidine/2-aminopyridines. This α,β-C-H functionalization is remarkable for its simplicity in both substrates and conditions, involving one and a half oxygen molecules and releasing three water molecules as the only byproduct. In addition, the functionalized products including protected 2-aminoimidazoles introduced without aromatization can serve as useful building blocks for natural product synthesis and medicinal chemistry.


Intratumoral angiogenesis: a new prognostic indicator for stage I endometrial adenocarcinomas?
The prognostic significance of three recently emerged parameters, namely intratumoral angiogenesis and the antiapoptotic proteins bcl-2 and mutant p53, was investigated in a series of 124 patients with endometrial adenocarcinomas of the endometrioid cell type. All patients were treated with total abdominal hysterectomy and bilateral oophorectomy, without node dissection. When deep myometrial invasion or advanced stage of disease was confirmed, adjuvant radiotherapy was given. Intratumoral angiogenesis was assessed in tissue samples, after immunohistochemical staining, with the anti-CD31 monoclonal antibody. The mean microvessel density (MVD) was 23.2 +/- 14.1 (range 4-60; 95% CI 20-25.8). Microvessel density was high (> 30) in 30% of endometrial adenocarcinomas, medium (15-30) in 33% of the tumors, and low (< 15) in the remaining cases (37%). A strong cytoplasmic and/or perinuclear expression of bcl-2 in more than 10% of the neoplastic cells was considered as being positive, and noted in 35.5% of the endometrial neoplasms; it was more frequent in the less vascularized carcinomas (P = 0.03). Nuclear p53 accumulation in an equal percentage of neoplastic cells (> 10%) was less common (7.2%). In univariate analysis, early stage of disease, absence of lymphatic-vascular space invasion (LVI), and low intratumoral MVD were the parameters associated with an improved survival (P = 0.0001, P = 0.001, and P = 0.009, respectively). In multivariate analysis, however, the only independent variable noted was stage of disease (P < 0.0001). Within stage I endometrial adenocarcinomas, only intratumoral angiogenesis was associated with prognosis (univariate analysis): high MVD cases had a significantly worse prognosis compared to medium MVD (P = 0.02). Low MVD adenocarcinomas, on the other hand, were associated with an intermediate prognosis, indicating that other factors, such as hypoxia and related mechanisms, may also be important. It is suggested that intratumoral angiogenesis may prove useful in selecting a subgroup of cancer patients, among others with stage I endometrial disease, that would benefit from additional treatment.


[Overweight among adolescents in Oslo].
Socio-demographic factors affect the prevalence of overweight, and differences in lifestyle could be the cause. Possible consequences of overweight, such as mental health problems may also vary by socio-demographic factors. The aims of the study were to identify overweight and mental health problems by socio-demographic group and lifestyle and a possible association between overweight and mental health. A questionnaire survey (UngHUBRO) was conducted among 10th graders in Oslo in 2000 and 2001. The response rate was 88% (N = 7343). Self-reported height and weight were used to determine overweight/obesity according to international standards. The prevalence of overweight/obesity and mental distress by socio-demographic group was calculated, and associations between the two were adjusted for lifestyle (dietary habits, physical activity and inactivity) by logistic regression. The prevalence of overweight/obesity was higher among boys (12%) than among girls (7%), and there was an inverse association with the length of parents' education for both genders. The association between overweight/obesity and mental distress was statistically significant for girls. The ethnic differences in overweight/obesity were large, but not statistically significant. The prevalence of overweight varied largely between socio-demographic groups. There was an association between mental distress and overweight among girls and there are marked differences in lifestyle between over- and normal weight adolescents.


The role of C-Fos protein, somatostatin and neuropeptide Y in the pathogenesis of ischemic brain injuries based on animal model of cerebral ischemia.
The aim of this study was to define all the areas of changes in expression of nuclear c-Fos protein (c-Fos), cytoplasmic somatostatin (SS) and neuropeptide Y (NPY) in rat brain during experimental ischemia. Using the immunohistochemical method, brain mapping (based on the atlas by Paxinos & Watson) of immunoreactivity for c-Fos, SS and NPY in 39 rats, was studied in telencephalon, diencephalon and midbrain after resistant and transitory ischemia. The first experimental group (R group) was exposed to resistant ischemia by occlusion (10 minutes) of four vessels according to the Pulsinelli method. The second group was first exposed to transitory (4 minutes) ischemia (preconditioning) and, after 72 hours, to total ischemia as in the R group. There was a statistical difference between the R and T group in the c-Fos reaction, especially in the parietofrontal cortex, anterior amygdaloid area, claustrum, reuniens nucleus and suprachiasmatic nucleus. The dominant immunohistochemical reactivity was found for c-Fos protein, and the most reactive in terms of co-localization of c-Fos with SS and NPY was periventricular area of hypothalamus. The mapping showed that both, phylogenetically new as well as phylogenetically older brain structures reacted immunohistochemically. The results of our study, regarding the impact of preconditioning with a short period of ischemia on c-Fos activity and co-localization of c-Fos with SS and NPY immunoreactivity, showed the need for future studies of brain neuropeptides related to regional and time effects, and indicated brain structures which may require pharmacological targeting to achieve neuroprotective level of proto-oncogene activity in populations at risk.


Canine parvovirus in New Zealand: epidemiological features and diagnostic methods.
Data from 763 cases of clinical canine parvovirus disease confirmed at the Ruakura Animal Health Laboratory were examined. The largest number of cases were seen in spring and summer months with the peak incidence in February 1981. The morbidity and mortality rates were highest in young dogs. Sixty-nine percent of all cases occurred in dogs less than six months of age, and 63 percent of dogs seven weeks of age or younger died. The laboratory methods used to diagnose canine parvovirus disease are compareh and discussed.


Modeling of coagulation, curd firming, and syneresis of milk from Sarda ewes.
This study investigated the modeling of curd-firming (CF) over time (CF(t)) of sheep milk. Milk samples from 1,121 Sarda ewes from 23 flocks were analyzed for coagulation properties. Lactodynamographic analyses were conducted for up to 60 min, and 240 CF individual observations from each sample were recorded. Individual sample CFt equation parameters (RCT(eq), rennet coagulation time; CF(P), asymptotic potential value of curd firmness; k(CF), curd-firming instant rate constant; and k(SR), curd syneresis instant rate constant) were estimated, and the derived traits (CF(max), the point at which CF(t) attained its maximum level, and tmax, the time at which CF(max) was attained) were calculated. The incidence of noncoagulating milk samples was 0.4%. The iterative estimation procedure applied to the individual coagulation data showed a small number of not-converged samples (4.4%), which had late coagulation and an almost linear pattern of the ascending part of the CF(t) curve that caused a high value of CF(P), a low value of k(CF), and a high value of k(SR). Converged samples were classified on the basis of their CF(t) curves into no-k(SR) (18.0%), low-k(SR) (72.6%), and high-k(SR) (4.5%). A CF(t) that was growing continuously because of the lack of the syneresis process characterized the no-k(SR) samples. The high-k(SR) samples had a much larger CFP, a smaller k(CF), and an anticipation of tmax, whereas the low-k(SR) samples had a fast k(CF) and a slower k(SR). The part of the average CF(t) curves that showed an increase was similar among the 3 different syneretic groups, whereas the part that decreased was different because of the expulsion of whey from the curd. The traditional milk coagulation properties recorded within 30 min were not able to detect any appreciable differences among the 4 groups of coagulating samples, which could lead to a large underestimation of the maximum CF of all samples (if predicted by a30), with the exception of the no-k(SR) samples. Large individual variability was found and was likely caused by the effects of the dairy system, such as flock size (on CF(max), t(max), and % ewes with no-k(SR) milk), flock within flock size (representing 11 to 43% of total variance for % ewes with no-k(SR) milk and CF(max), respectively), days in milk (on all model parameters and CF(max)), parity (on RCT(eq), k(SR), and CF(max)), daily milk yield (on RCT(eq) and CF(max)), and position of the individual pendulum that significantly affected model parameters and derived traits. In conclusion, the results showed that the modeling of coagulation, curd-firming, and syneresis is a suitable tool to achieve a deeper interpretation of the coagulation and curd-firming processes of sheep milk and also to study curd syneresis.


NUDT15 Polymorphism Confer Increased Susceptibility to Thiopurine-Induced Leukopenia in Patients With Autoimmune Hepatitis and Related Cirrhosis.
The aim of this study was to investigate the influence of NUDT15 R139C and thiopurine S-methyltransferase (TPMT) on azathioprine (AZA) induced leukopenia in patients with autoimmune hepatitis (AIH) and related cirrhosis. A total of 149 Chinese AIH patients with a history of AZA treatment were retrospectively evaluated. The clinical and epidemiological characteristics of the patients were obtained from an electronic database and reviewed. NUDT15 (rs116855232) and TPMT∗3C (rs1142345) SNPs were genotyped using a PCR method. Twelve patients developed leukopenia, and this adverse drug reaction was significantly associated with the T risk allele in NUDT15 [P < 0.00001, odds ratio = 20.41; 95% confidence interval (CI) (7.84, 53.13)], with the sensitivity and specificity of 91.67 and 89.05%, respectively. The median maintenance dosages for patients with the rs116855232 CC and CT genotypes were 1.23 (0.95, 1.53) mg ⋅ kg-1 ⋅ d-1 and 0.96 (0.83, 1.19) mg ⋅ kg-1 ⋅ d-1, respectively (P = 0.028). In contrast, no significant association was observed for TPMT∗3C genotypes. Notably, subgroup analysis of the 13 patients with leukopenia before therapy, these white blood cell (WBC) counts did not show further reduction after AZA treatment and maintenance dosage was 1.13 (0.94, 1.60) mg ⋅ kg-1 ⋅ d-1. Therefore, NUDT15 polymorphism is significantly associated with thiopurine-induced leukopenia in Chinese patients with AIH and related cirrhosis. Adjusting the AZA dosage should be considered in patients according to the NUDT15 R139C genotypes.


Removal of chronic Mycoplasma ovipneumoniae carrier ewes eliminates pneumonia in a bighorn sheep population.
Chronic pathogen carriage is one mechanism that allows diseases to persist in populations. We hypothesized that persistent or recurrent pneumonia in bighorn sheep (Ovis canadensis) populations may be caused by chronic carriers of Mycoplasma ovipneumoniae (Mo). Our experimental approach allowed us to address a conservation need while investigating the role of chronic carriage in disease persistence.We tested our hypothesis in two bighorn sheep populations in South Dakota, USA. We identified and removed Mo chronic carriers from the Custer State Park (treatment) population. Simultaneously, we identified carriers but did not remove them from the Rapid City population (control). We predicted removal would result in decreased pneumonia, mortality, and Mo prevalence. Both population ranges had similar habitat and predator communities but were sufficiently isolated to preclude intermixing.We classified chronic carriers as adults that consistently tested positive for Mo carriage over a 20-month sampling period (n = 2 in the treatment population; n = 2 in control population).We failed to detect Mo or pneumonia in the treatment population after chronic carrier removal, while both remained in the control. Mortality hazard for lambs was reduced by 72% in the treatment population relative to the control (CI = 36%, 91%). There was also a 41% reduction in adult mortality hazard attributable to the treatment, although this was not statistically significant (CI = 82% reduction, 34% increase). Synthesis and Applications: These results support the hypothesis that Mo is a primary causative agent of persistent or recurrent respiratory disease in bighorn sheep populations and can be maintained by a few chronic carriers. Our findings provide direction for future research and management actions aimed at controlling pneumonia in wild sheep and may apply to other diseases.


Advances in pattern formation: Signaling and transcription.
The 49(th) Annual Drosophila Research Conference was held in the sunny confines of San Diego. As usual, large numbers of Drosophila scientists working in fields as different as immunology and evolution descended on the venue. The meeting showed that the fly community is still vibrant and diverse even with the funding crunch at the NIH and the renewed rumors that Drosophila may have outlived its usefulness. This short review will focus on one session of platform presentations detailing the recent advances in the field of pattern formation. This session offered a variety of topics reviewing the formation of pattern in various tissues through diverse mechanisms. I will focus on early embryonic patterning through pair-rule genes, specificity of FGF signaling, and tissue regeneration.


A randomized, double-blind, comparative trial of a new oral combination of artemether and benflumetol (CGP 56697) with mefloquine in the treatment of acute Plasmodium falciparum malaria in Thailand.
CGP 56697, a new oral fixed combination of artemether and benflumetol, was tested in a double-blinded, randomized trial in 252 adult patients treated either with CGP 56697 (4 x 4 tablets each containing 20 mg of artemether and 120 mg of benflumetol, given at 0, 8, 24, and 48 hr), or with mefloquine (three tablets of 250 mg at initial diagnosis, followed by two tablets of 250 mg at 8 hr). Baseline data of the two groups were comparable. The 28-day cure rate with CGP 56697 was lower than with mefloquine (69.3% versus 82.4%; P = 0.002). However, CGP 56697 was more effective than mefloquine in parasite clearance time (43 hr versus 66 hr; P < 0.001) fever clearance time (32 hr versus 54 hr; P < 0.005), and gametocyte clearance time (152 hr versus 331 hr; P < 0.001). This study revealed that CGP 56697 is effective against multidrug-resistant Plasmodium falciparum malaria in Thailand, but higher doses will probably be needed to improve the cure rate.


Hypoglycemic and hypocholesterolemic activity of leave of few medicinal plants against steptozotocin induced hyperglycemia.
Diabetes mellitus (DM) is universal of the hormonal problem and Type II diabetes is foremost obstacle. Accessible management in medicine has numerous contrary paraphernalia. Medical flora shows an essential part in managing diabetes specifically in unindustrialized nations. The present study was done on leaves of Rhazya stricta Decane, Adhatoda zeylanica, Berberis lycium Royle and Olea furrrignea, whose methanolic extracts were used to check their hypoglycemic and hypolipidemic activity by using glucometer and kit method respectively in blood of male and female albino mice Balb C. Results showed that leaves of R. stricta were best for hypoglycemia (125.34±63.79mg/dl, 107.34±18.00mg/dl, 146.00±40.36mg/dl and 178.34±17.03mg/dl), hypocholesterolemia (147.88±21.83mg/dl and 125.89±14.03mg/dl) and triglycerides (103±8.88mg/dl and 89.±43.4mg/dl) in random and fasting conditions, in male and female mice respectively. All plant extract were most effective for hypoglycemia and hypocholesterolemia in female mice as compared to male mice. Moreover statistical analysis revealed that leaves of other plants were also effective but less than leaves of R. stricta. So this plant part and a combination of presently used plants can be used for further studies particularly among females by the purification of active compounds against hyperglycemia and hypercholesterolemia.


Phase II trial of bevacizumab in recurrent or persistent endometrial cancer: a Gynecologic Oncology Group study.
Bevacizumab, a recombinant humanized monoclonal antibody against vascular endothelial growth factor-A (VEGF-A), has clinical activity in multiple tumor types. We conducted a phase II trial to assess the activity and tolerability of single-agent bevacizumab in recurrent or persistent endometrial cancer (EMC). Eligible patients had persistent or recurrent EMC after receiving one to two prior cytotoxic regimens, measurable disease, and Gynecologic Oncology Group performance status of ≤ 2. Treatment consisted of bevacizumab 15 mg/kg intravenously every 3 weeks until disease progression or prohibitive toxicity. VEGF-A was assessed by immunohistochemistry in archival tumor and by enzyme-linked immunosorbent assay in pretreatment plasma. Primary end points were progression-free survival (PFS) at 6 months and overall response rate. Fifty-six patients were enrolled. Fifty-two patients were eligible and evaluable. Median age was 62 years, and prior treatment consisted of one or two regimens in 33 (63.5%) and 19 (36.5%) patients, respectively. Twenty-nine patients (55.8%) received prior radiation. Adverse events were consistent with those expected with bevacizumab treatment. No GI perforations or fistulae were seen. Seven patients (13.5%) experienced clinical responses (one complete response and six partial responses; median response duration, 6.0 months), and 21 patients (40.4%) survived progression free for at least 6 months. Median PFS and overall survival times were 4.2 and 10.5 months, respectively. Suggested associations were observed between high VEGF-A and adjusted hazard of death or tumor response when evaluated in tumor/plasma or plasma, respectively. Bevacizumab is well tolerated and active based on PFS at 6 months in recurrent or persistent EMC and warrants further investigation.


Subdural empyma due to Mycobacterium fortuitum in a non-HIV patient.
A 14-year-old male child presented with high grade intermittent fever with altered sensorium since 5-6 days and generalised seizures. On examination neck stiffness noticed with normal haemogram and chest X-ray. CSF microscopy was normal and no growth seen in aerobic culture. CT scan showed loculated lesion. Drained pus showed acid fast organism and culture on Lowestein Jensen medium showed pale-coloured growth on 3 rd day. Organism identified as Mycobacterium fortuitum by biochemical test. Interesting aspect of this case was there is no history of trauma or injection and patient was negative for HIV antibody.


Analysis of site performance in academic-based and community-based centers in the IDEAL Study.
To evaluate differences in metrics of quality and site performance in academic and community sites participating in a multicenter study. In the Individualized Dosing Efficacy Versus Flat Dosing to Assess Optimal Pegylated Interferon Therapy study, the participation of 76 academic-based and 42 community-based US centers provided an opportunity to evaluate various metrics of quality and site performance. A secondary data analysis of the Individualized Dosing Efficacy Versus Flat Dosing to Assess Optimal Pegylated Interferon Therapy study was performed. There were 3070 treatment-naive, hepatitis C virus genotype 1 infected patients were included. We retrospectively evaluated rates of screen failure, completion, and discontinuation of treatment and follow-up, treatment adherence, and virologic response by site type. Of the patients screened, 63% and 37% were in academic and community centers, respectively. Screen failure rates were similar (30% to 32%). End-of-treatment response, relapse, and sustained virologic response (SVR) rates in academic and community centers did not differ. SVR was achieved in 40% of patients at academic sites and 39% at community sites. Adherence to ≥80% of peginterferon-α and ribavirin dosing for ≥80% assigned duration was also similar (46% in academic and 47% in community centers). In both academic and community centers, 54% of patients completed treatment; there were similar discontinuation rates for treatment failure and adverse events. There were no significant differences in adherence, adverse events, rates of discontinuation, on-treatment virologic response, and SVR when comparing academic and community sites. The performance of academic-based and experienced community-based sites in clinical trials is largely similar for the treatment of chronic hepatitis C.


The environmental health of Latino children.
Representing 1 in 6 children in the United States, Latino children incur disproportionate exposures to air pollutants, pesticides, and toxic industrial chemicals, as well as lead and mercury from candy, traditional folk remedies, religious practices, and other sources. Latino children also have higher rates of asthma, lead and mercury poisoning, behavioral and developmental disorders, and certain cancers. Concurrent exposure to multiple pollutants, pre-existing disease, poor nutrition, substandard housing, limited access to health care, and other factors related to their lower socioeconomic status increase Latino children's susceptibility to environmental contaminants. Targeted research, education, prevention and intervention efforts, and economic development initiatives are needed.


Therapeutic plasma exchange reduces ABO titers to permit ABO-incompatible renal transplantation.
Thousands of patients with chronic renal failure die yearly without a kidney transplant due to the severe shortage of donors. Therapeutic plasma exchange (TPE) is performed to permit ABO-incompatible (ABO-I) kidney transplants, but little is known about how well TPE reduces ABO antibodies or complications related to TPE in this clinical setting. This retrospective study evaluated 46 individuals that received TPE to permit ABO-I kidney transplant. The number of TPE treatments was based on a goal ABO titer at the anti-human globulin (AHG) phase of 16 or less before surgery. Before TPE, the median titer of recipient was 32 (range, 2-128) at room temperature (RT) phase and 64 (range, 4-1024) at AHG phase. The first TPE reduced the total agglutination reactivity score at AHG phase by 10.2 percent. Before transplantation, there was a mean of 6.2 +/- 2.5 TPE treatments and total agglutination reactivity score at AHG phase was reduced by 53.5 percent. The median titer remained reduced at 3 to 6 months after transplantation at 4 (range, 0-64) at RT phase and 8 (range, 1-64) at AHG phase. TPE complications were minimal. During at least one procedure, 15 (32.6%) individuals had either urticaria or pruritus, 18 (39.1%) individuals experienced mild citrate-induced hypocalcemia, 5 (10.2%) individuals had hypotension, 6 (13.0%) individuals had nausea or vomiting, and 1 (2.2%) individual had West Nile virus encephalitis. With current infectious disease blood screening protocols, TPE has minimal complications and can reduce ABO antibody titers to permit ABO-I renal transplantation.


Adenovirus-mediated p53 gene therapy inhibits human sarcoma tumorigenicity.
Mutations of the p53 tumor-suppressor gene are the most frequent genetic abnormality in soft tissue sarcomas. Because these rare tumors also respond poorly to standard chemotherapy and bear a 50% 5-year mortality rate, we investigated the possible therapeutic benefits of p53 gene restoration in sarcomas. We constructed Ad5p53, which is an E1A-deleted, replication-deficient adenovirus expressing a cytomegalovirus promoter-driven wild-type p53 cDNA with a Flag sequence tag. SKLMS-1 human leiomyosarcoma cells containing a mis-sense p53 point mutation were effectively transduced with Ad5p53. Increasing levels of Flag-p53 protein, as well as dose-dependent p21Cip1 induction, were observed through a dose range of 10-500 plaque-forming units (PFU)/cell. In vitro administration of Ad5p53 as a single 100 PFU/cell dose caused 40-60% growth inhibition of SKLMS-1 cells at posttreatment days 4, 6, and 8 compared with untreated or viral control treated-cells (P < .05, Student's t test). Relative to these same controls, in vivo treatment of SKLMS-1-bearing severe combined immunodeficient mice with 6 x 10(9) PFU of Ad5p53 by intratumoral injection resulted in a 35-day tumor growth delay and complete tumor regression in 40% of mice (P < .05, Student's t test). The expression of virally derived p53 mRNA in Ad5p53-treated tumor tissues was detected in treated tumor specimens by reverse transcriptase polymerase chain reaction. Reduced intratumoral cellularity and the presence of p53 staining in adjacent normal tissue, consistent with delivery of exogenous p53 to the tumor target, were evident only in Ad5p53-treated tumors after immunohistochemical staining for p53. These results indicate that wild-type p53 gene restoration in sarcomas retards tumor growth and may come to be usefully applied to the clinical treatment of this disease as a single regimen or in combination with conventional therapies.


The relationship between hippocampal asymmetry and temperament in adolescent borderline and antisocial personality pathology.
Investigating etiological processes early in the life span represents an important step toward a better understanding of the development of personality pathology. The current study evaluated the interaction between an individual difference risk factor (i.e., temperament) and a biological risk factor for aggressive behavior (i.e., atypical [larger] rightward hippocampal asymmetry) in predicting the emergence of borderline personality disorder (BPD) and antisocial personality disorder symptoms during early adolescence. The sample consisted of 153 healthy adolescents (M = 12.6 years, SD = 0.4, range = 11.4-13.7) who were selected from a larger sample to maximize variation in temperament. Interactions between four temperament factors (effortful control, negative affectivity, surgency, and affiliativeness), based on the Early Adolescent Temperament Questionnaire-Revised, and volumetric measures of hippocampal asymmetry were examined as cross-sectional predictors of BPD and antisocial personality disorder symptoms. Boys were more likely to have elevated BPD symptoms if they were high on affiliation and had larger rightward hippocampal asymmetry. In boys, low affiliation was a significant predictor of BPD symptoms in the presence of low rightward hippocampal asymmetry. For girls, low effortful control was associated with elevated BPD symptoms in the presence of atypical rightward hippocampal asymmetry. This study builds on previous work reporting significant associations between atypical hippocampal asymmetry and poor behavioral regulation.


Effects of dDAVP on rat juxtamedullary nephrons: stimulation of medullary K recycling.
The effects of 1-desamino-8-D-arginine vasopressin (dDAVP) on the handling of water and electrolytes by the juxtamedullary nephrons were studied on rats with reduced circulating levels of antidiuretic hormone (ADH), parathyroid hormone, calcitonin, and glucagon, all of which stimulate the adenylate cyclase system of the thick ascending limb and the distal tubule. In such hormone-deprived rats and in hormone-deprived + dDAVP rats, the concentration of Na, Cl, and total solutes was lower in the ascending than in the descending limbs, whereas the inulin concentration was similar at both sites. dDAVP did not alter the fraction of NaCl remaining in the thin limbs, but tended to reduce that of Mg and Ca. On the other hand, dDAVP significantly increased the fraction of filtered K remaining from 65.8 +/- 5.2 to 107.3 +/- 15.8%. A direct correlation was observed between the fraction of filtered K remaining at the tip of the juxtamedullary loops and the fractional excretion rate of K in urine. Since dDAVP enhances distal K net secretion, as previously shown in our laboratory, these results indicate that the medullary recycling of K from nephron terminal segments to Henle's loop of juxtamedullary nephrons is stimulated by this peptide.


Catalase and superoxide dismutase conjugated with platelet-endothelial cell adhesion molecule antibody distinctly alleviate abnormal endothelial permeability caused by exogenous reactive oxygen species and vascular endothelial growth factor.
Reactive oxygen species (ROS) superoxide anion (O(2)()) and hydrogen peroxide (H(2)O(2)) produced by activated leukocytes and endothelial cells in sites of inflammation or ischemia cause endothelial barrier dysfunction that may lead to tissue edema. Antioxidant enzymes (AOEs) catalase and superoxide dismutase (SOD) conjugated with antibodies to platelet-endothelial cell adhesion molecule-1 (PECAM-1) specifically bind to endothelium, quench the corresponding ROS, and alleviate vascular oxidative stress and inflammation. In the present work, we studied the effects of anti-PECAM/catalase and anti-PECAM/SOD conjugates on the abnormal permeability manifested by transendothelial electrical resistance decline, increased fluorescein isothiocyanate-dextran influx, and redistribution of vascular endothelial-cadherin in human umbilical vein endothelial cell (HUVEC) monolayers. Anti-PECAM/catalase protected HUVEC monolayers against H(2)O(2)-induced endothelial barrier dysfunction. Polyethylene glycol-conjugated catalase exerted orders of magnitude lower endothelial uptake and no protective effect, similarly to IgG/catalase. Anti-PECAM/catalase, but not anti-PECAM/SOD, alleviated endothelial hyperpermeability caused by exposure to hypoxanthine/xanthine oxidase, implicating primarily H(2)O(2) in the disruption of the endothelial barrier in this model. Thrombin-induced endothelial permeability was not affected by treatment with anti-PECAM/AOEs or the NADPH oxidase inhibitor apocynin or overexpression of AOEs, indicating that the endogenous ROS play no key role in thrombin-mediated endothelial barrier dysfunction. In contrast, anti-PECAM/SOD, but not anti-PECAM/catalase, inhibited a vascular endothelial growth factor (VEGF)-induced increase in endothelial permeability, identifying a key role of endogenous O(2)() in the VEGF-mediated regulation of endothelial barrier function. Therefore, AOEs targeted to endothelial cells provide versatile molecular tools for testing the roles of specific ROS in vascular pathology and may be translated into remedies for these ROS-induced abnormalities.


High-resolution analysis of central nervous system expression patterns in zebrafish Gal4 enhancer-trap lines.
The application of the Gal4/UAS system to enhancer and gene trapping screens in zebrafish has greatly increased the ability to label and manipulate cell populations in multiple tissues, including the central nervous system (CNS). However the ability to select existing lines for specific applications has been limited by the lack of detailed expression analysis. We describe a Gal4 enhancer trap screen in which we used advanced image analysis, including three-dimensional confocal reconstructions and documentation of expression patterns at multiple developmental time points. In all, we have created and annotated 98 lines exhibiting a wide range of expression patterns, most of which include CNS expression. Expression was also observed in nonneural tissues such as muscle, skin epithelium, vasculature, and neural crest derivatives. All lines and data are publicly available from the Zebrafish International Research Center (ZIRC) from the Zebrafish Model Organism Database (ZFIN). Our detailed documentation of expression patterns, combined with the public availability of images and fish lines, provides a valuable resource for researchers wishing to study CNS development and function in zebrafish. Our data also suggest that many existing enhancer trap lines may have previously uncharacterized expression in multiple tissues and cell types.


Parasites in food webs: the ultimate missing links.
Parasitism is the most common consumer strategy among organisms, yet only recently has there been a call for the inclusion of infectious disease agents in food webs. The value of this effort hinges on whether parasites affect food-web properties. Increasing evidence suggests that parasites have the potential to uniquely alter food-web topology in terms of chain length, connectance and robustness. In addition, parasites might affect food-web stability, interaction strength and energy flow. Food-web structure also affects infectious disease dynamics because parasites depend on the ecological networks in which they live. Empirically, incorporating parasites into food webs is straightforward. We may start with existing food webs and add parasites as nodes, or we may try to build food webs around systems for which we already have a good understanding of infectious processes. In the future, perhaps researchers will add parasites while they construct food webs. Less clear is how food-web theory can accommodate parasites. This is a deep and central problem in theoretical biology and applied mathematics. For instance, is representing parasites with complex life cycles as a single node equivalent to representing other species with ontogenetic niche shifts as a single node? Can parasitism fit into fundamental frameworks such as the niche model? Can we integrate infectious disease models into the emerging field of dynamic food-web modelling? Future progress will benefit from interdisciplinary collaborations between ecologists and infectious disease biologists.


Strengthening local governance in health financing in China: a text-mining analysis of policy changes between 2009 and 2020.
In the last two decades, developing countries have increasingly engaged in improving the governance of their health systems and promoting policy design to strengthen their health governance capacity. Although many well-designed national policy strategies have been promulgated, obstacles to policy implementation and compliance among localities may undermine these efforts, particularly in decentralized health systems. Studies on health governance have rarely adopted a central-local analysis to investigate in detail local governments' distinct experiences, orientations and dynamics in implementing the same national policy initiative. This study examines the policy orientations of prefectural governments in strengthening governance in health financing in China, which has transitioned from emphasizing the approach of fiscal resource input to that of marketization promotion and cost-containment regulation enforcement at the national level since 2009. Employing text-mining methodologies, we analysed health policy documents issued by multi-level governments after 2009. The analysis revealed three salient findings. Firstly, compared to higher-level authorities, prefectural governments generally opted to use fiscal resource input over marketization promotion and cost-containment regulation enforcement between 2009 and 2020. Secondly, policy choices of prefectural governments varied considerably in terms of enforcing cost-containment regulations during the same period. Thirdly, the extent of the prefectural government's orientation toward marketization promotion or cost-containment regulation enforcement was not only determined by the top-down orders of higher-level authorities but was also incentivized by the government's fiscal dependency and the policy orientations of peer governments. These findings contribute to the health governance literature by providing an overview of local discretion in policy choices and the political and fiscal dynamics of local policy orientations in promoting health governance in a decentralized health system.


Control of formation and dissociation of the high-affinity complex between cytochrome c and cytochrome c peroxidase by ionic strength and the low-affinity binding site.
A new ruthenium photoreduction technique was used to measure the formation and dissociation rate constants kf and kd of the high-affinity complex between yeast iso-1-cytochrome c (yCc) and cytochrome c peroxidase compound I (CMPI) over a wide range of ionic strength. These studies utilized Ru-39-Cc, which contains trisbipyridylruthenium attached to the cysteine residue in the H39C, C102T variant of yCc, and has the same reactivity with CMPI as native yCc. kd and kf were measured by photoreducing a small concentration of Ru-39-Cc in the presence of the oxidized yCcIII: CMPI complex, which must dissociate before Ru-39-CcII can bind to CMPI and reduce the radical action. The value of kd for the 1:1 high-affinity complex is very small at low ionic strength, < 5 s-1 but is increased significantly by binding yCc to a second low-affinity site. However, the low-affinity yCc binding site is not active in direct electron transfer to either the radical cation or the oxyferryl heme in CMPI, and is too weak to play a role in the kinetics at ionic strengths above 70 mM. The value of kd increases to 4000 s-1 at 150 mM ionic strength, while kf decreases from > 3 x 10(9) M-1 s-1 at low ionic strength to 1.3 x 10(9) M-1 s-1 at 150 mM ionic strength. These studies indicate that the rate-limiting step in enzyme turnover is product dissociation below 150 mM ionic strength and intracomplex electron transfer to the oxyferryl heme at higher ionic strength. The interaction between yCc and CcP is optimized at physiological ionic strength to provide the largest possible complex formation rate constant kf without allowing product dissociation to be rate-limiting. The effects of surface mutations on the kinetics provided evidence that the high-affinity binding site used for the reaction in solution is similar to the one identified in the yCc:CcP crystal structure.


Child health: evidence on the roles of family income and maternal mental health from a UK birth cohort.
There is a growing literature showing an association between higher family income and better child health. This paper uses cohort data with rich information on mother's early life events, her health, child-health-related behaviours, and her child's health to examine this association for the UK and to identify some of the mechanisms through which income affects child health. The paper examines the cross-sectional association between income and health, finds the expected association, but concludes that the association with current income cannot be distinguished from one between permanent income and child health. It then focuses on the mechanisms by which income translates into better child health; these include parental behaviours that may affect child health and parental health, including maternal mental health. Controlling for these factors, there is almost no direct impact of income. A significant role is played by mother's own health, particularly her mental health. No clear role is played by child-health production behaviours of the mother. Examining the maternal mental health-child health link in more detail suggests a role for maternal anxiety and somaticism.


Adaptive Resistance to Chemotherapy, A Multi-FAK-torial Linkage.
Oncogenes provide tumor cells with a growth and survival advantage. Directed therapies targeted to oncogenic mutations (such as BRAF V600E) are part of effective late-stage melanoma treatment. However, tumors with BRAF V600E mutations, in approximately 10% of colorectal cancer, are generally treatment-insensitive. Research has identified various "feedback" mechanisms that result in BRAF signal pathway reactivation in response to BRAF inhibition. Herein, we highlight key findings from Chen and colleagues (this issue) showing that integrin-associated focal adhesion kinase (FAK) activation selectively occurs in BRAF V600E-mutant colorectal cancer cells in response to pharmacological BRAF inhibition. FAK activation results in elevated β-catenin protein levels, β-catenin nuclear localization, and increased gene transcription. Small-molecule inhibitors of β-catenin or FAK synergize with vemurafenib BRAF inhibitor to prevent BRAF V600E colorectal cancer cell proliferation in vitro and xenograft tumor growth in mice. This study complements findings linking FAK to β-catenin in intestinal tumorigenesis, resistance to radiotherapy, and cancer stem cell survival. Thus, FAK activation may occur as a frequent tumor cell "adaptive resistance" mechanism. Although FAK (PTK2) is not mutated in most cancers, targeting FAK activity in combinational approaches may limit tumor cell escape mechanisms and enhance durable responses to treatment. Mol Cancer Ther; 17(4); 719-23. ©2018 AACR.


Boronic Acid Appended Naphthyl-Pyridinium Receptors as Chemosensors for Sugars.
There remains a need in clinics and research to have simple and sensitive detection systems that allow the detection and quantification of sugar markers of biomedical relevance such as sugars lactulose and mannitol for noninvasive gut permeability assessment. We have prepared a new class of boronic acid-appended naphthyl-pyridinium receptor compounds as chemosensors. These were studied for their ability to act as modular internal charge transfer (ICT) fluorescent probes or donor/acceptor pair ensembles where the receptor compound can act as a quencher for an anionic dye. As an ICT sensor, fluorescence intensity increased upon diol recognition, which stems from the neutralization of the pyridinium nitrogen that is perturbing the chromophoric properties. We found these ICT probes provide good sensitivity for disaccharide lactulose with low micromolar detection and quantification limits. In addition, their ability to form a non-fluorescent ground state complex with anionic reporter dyes, such as HPTS or TSPP, was examined as probes for various sugars. We have identified three receptor/quencher compounds with high quenching efficiency for anionic dyes. Subsequently, a range of sugars and sugar derivatives were tested for chemosenstivity of our probes. This study illustrates an approach for designing boronic acid-based chemoreceptors for the recognition and quantification of sugars and sugar derivatives.


Arthroscopic debridement of intercarpal ligament and triangular fibrocartilage complex tears.
Wrist arthroscopy has evolved since its inception to become an essential diagnostic and therapeutic tool for the management of various wrist disorders. Our study aimed to examine the outcomes of arthroscopic debridement in the treatment of scapholunate (SL), lunotriquetral (LT) and triangular fibrocartilage complex (TFCC) tears. We conducted a retrospective review of 68 consecutive wrist arthroscopies performed at our institution between January 2000 and July 2005. All the patients complained of wrist pain, which often interfered with their daily activities, work or sports. A standard arthroscopic technique was employed in all. Any intercarpal ligament or TFCC tears found were debrided. There were 42 patients with TFCC tears, 58 with SL tears and 49 with LT tears. At a mean follow-up time of 16.6 months, 85.3% of the patients reported an improvement in symptoms and 27.9% had improved range of motion. Grip strength improved by 11.8%. All except two patients returned to their original activities. Outcome following arthroscopic debridement was determined using the Mayo Modified Wrist Score. Based on the postoperative wrist scores of 47 patients, 24 were rated excellent, 17 good, four fair and two poor. By comparing the pre- and postoperative wrist scores of 31 patients, we were able to demonstrate significant improvement in patients who underwent wrist arthroscopies. Our study demonstrated that there is a definite role for arthroscopic debridement in the management of SL, LT and TFCC tears.


ANN-Spec: a method for discovering transcription factor binding sites with improved specificity.
This work describes ANN-Spec, a machine learning algorithm and its application to discovering un-gapped patterns in DNA sequence. The approach makes use of an Artificial Neural Network and a Gibbs sampling method to define the Specificity of a DNA-binding protein. ANN-Spec searches for the parameters of a simple network (or weight matrix) that will maximize the specificity for binding sequences of a positive set compared to a background sequence set. Binding sites in the positive data set are found with the resulting weight matrix and these sites are then used to define a local multiple sequence alignment. Training complexity is O(lN) where l is the width of the pattern and N is the size of the positive training data. A quantitative comparison of ANN-Spec and a few related programs is presented. The comparison shows that ANN-Spec finds patterns of higher specificity when training with a background data set. The program and documentation are available from the authors for UNIX systems.


Adenoviral gene delivery for HIV-1 vaccination.
The AIDS epidemic continues to spread throughout nations of Africa and Asia and is by now threatening to undermine the already frail infrastructure of developing countries in Sub-Saharan Africa that are hit the hardest. The only option to stem this epidemic is through inexpensive and efficacious vaccines that prevent or at least blunt HIV-1 infections. Despite decades of pre-clinical and clinical research such vaccines remain elusive. Most anti-viral vaccines act by inducing protective levels of virus-neutralizing antibodies. The envelope protein of HIV-1, the sole target of neutralizing antibodies, is constantly changing due to mutations, B cell epitopes are masked by heavy glycosylation and the protein's structural unfolding upon binding to its CD4 receptor and chemokine co-receptors. Efforts to induce broadly cross-reactive virus-neutralizing antibodies able to induce sterilizing or near sterilizing immunity to HIV-1 have thus failed. Studies have indicated that cell-mediated immune responses and in particular CD8+ T cell responses to internal viral proteins may control HIV-1 infections without necessarily preventing them. Adenoviral vectors expressing antigens of HIV-1 are eminently suited to stimulate potent CD8+ T cell responses against transgene products, such as antigens of HIV-1. They performed well in pre-clinical studies in rodents and nonhuman primates and are currently in human clinical trials. This review summarizes the published literature on adenoviral vectors as vaccine carriers for HIV-1 and discusses advantages and disadvantages of this vaccine modality.


Recessive missense LAMP3 variant associated with defect in lamellar body biogenesis and fatal neonatal interstitial lung disease in dogs.
Neonatal interstitial lung diseases due to abnormal surfactant biogenesis are rare in humans and have never been reported as a spontaneous disorder in animals. We describe here a novel lung disorder in Airedale Terrier (AT) dogs with clinical symptoms and pathology similar to the most severe neonatal forms of human surfactant deficiency. Lethal hypoxic respiratory distress and failure occurred within the first days or weeks of life in the affected puppies. Transmission electron microscopy of the affected lungs revealed maturation arrest in the formation of lamellar bodies (LBs) in the alveolar epithelial type II (AECII) cells. The secretory organelles were small and contained fewer lamellae, often in combination with small vesicles surrounded by an occasionally disrupted common limiting membrane. A combined approach of genome-wide association study and whole exome sequencing identified a recessive variant, c.1159G>A, p.(E387K), in LAMP3, a limiting membrane protein of the cytoplasmic surfactant organelles in AECII cells. The substitution resides in the LAMP domain adjacent to a conserved disulfide bond. In summary, this study describes a novel interstitial lung disease in dogs, identifies a new candidate gene for human surfactant dysfunction and brings important insights into the essential role of LAMP3 in the process of the LB formation.


Microleakage and the class V composite cavosurface.
The following conclusions may be made from this study: Substantial leakage occurs around unetched composite restorations. No procedure employed in this study consistently eliminated all leakage. Etching the peripheral enamel decreased penetration of the dye. The use of an intermediary bonding resin did not enhance marginal sealing over that obtained when only the etchant was used. The gingival margin was less resistant to penetration of the dye than the occlusal margin. Beveling the cavosurface margin produced a significant decrease in leakage, when employed with acid etching in Group II.


Philicities, Fugalities, and Equilibrium Constants.
The mechanistic model of Organic Chemistry is based on relationships between rate and equilibrium constants. Thus, strong bases are generally considered to be good nucleophiles and poor nucleofuges. Exceptions to this rule have long been known, and the ability of iodide ions to catalyze nucleophilic substitutions, because they are good nucleophiles as well as good nucleofuges, is just a prominent example for exceptions from the general rule. In a reaction series, the Leffler-Hammond parameter α = δΔG(⧧)/δΔG° describes the fraction of the change in the Gibbs energy of reaction, which is reflected in the change of the Gibbs energy of activation. It has long been considered as a measure for the position of the transition state; thus, an α value close to 0 was associated with an early transition state, while an α value close to 1 was considered to be indicative of a late transition state. Bordwell's observation in 1969 that substituent variation in phenylnitromethanes has a larger effect on the rates of deprotonation than on the corresponding equilibrium constants (nitroalkane anomaly) triggered the breakdown of this interpretation. In the past, most systematic investigations of the relationships between rates and equilibria of organic reactions have dealt with proton transfer reactions, because only for few other reaction series complementary kinetic and thermodynamic data have been available. In this Account we report on a more general investigation of the relationships between Lewis basicities, nucleophilicities, and nucleofugalities as well as between Lewis acidities, electrophilicities, and electrofugalities. Definitions of these terms are summarized, and it is suggested to replace the hybrid terms "kinetic basicity" and "kinetic acidity" by "protophilicity" and "protofugality", respectively; in this way, the terms "acidity" and "basicity" are exclusively assigned to thermodynamic properties, while "philicity" and "fugality" refer to kinetics. Benzhydrylium ions (diarylcarbenium ions) with para- and meta-substituents are used as reference compounds for these investigations, because their Lewis acidities and electrophilicities can be varied by many orders of magnitude, while the steric surroundings of the reaction centers are kept constant. The rate constants for their reactions with nucleophiles correlate linearly over a wide range with the Lewis acidities of the benzhydrylium ions: from slow reactions with late transition states to very fast reactions with early, reactant-like transition states (including reactions which proceed without an enthalpic barrier, ΔH(⧧) = 0). Thus, unequivocal evidence is obtained that even within a series of closely related reactions, the Leffler-Hammond α cannot be a measure for the position of the transition state. Differences in intrinsic barriers lead to deviations from the linear rate-equilibrium correlations and give rise to counterintuitive phenomena. Thus, 1,4-diazabicyclo[2.2.2]octane (DABCO) reacts with lower intrinsic barriers than 4-(dimethylamino)pyridine (DMAP) and, therefore, is a stronger nucleophile as well as a better nucleofuge than DMAP. Common synthetically used SN2 reactions are presented, in which weak nucleophiles replace stronger ones. Whereas solvolysis rates of alkoxy- and alkyl-substituted benzhydryl derivatives correlate linearly with the Lewis acidities of the resulting carbenium ions, this is not the case for amino-substituted benzhydrylium ions, where differences in intrinsic barriers play a major role. The common rule that a structural variation, which increases the electrophilicity of a carbocation at the same time reduces its electrofugality, does not hold any longer. The need to systematically analyze the role of intrinsic barriers is emphasized.


[Alpha interferon, antiviral proteins and their value in clinical medicine].
Type I interferon system is an important part of host's innate defense mechanisms against viral infections. The type I interferons mediate in part their antiviral effect via induction of various proteins. Among them the most widely known are 2'-5' oligoadenylate synthetase (2'-5' OAS) and a protein kinase (PKR). MxA, an other antiviral protein, is specifically induced by the type I interferons. The MxA protein contains the dynamin signature, which is implicated in transport processes. The MxA protein appears to block the replication of certain viruses at poorly defined steps. There are substantial differences in the antiviral activity of MxA between virus types. Indeed, the replication of vesicular stomatitis virus and influenza virus is inhibited by MxA, but not the one of type I herpes simplex virus. Measurements of interferon alpha and MxA levels may be of high value in clinical practice. Interferon alpha can be detected by using a bioassay based on the interferon alpha ability to protect cultured cells from the cytopathic effect caused by a selected challenged virus, or by using immunological techniques. The current bioassays are the most sensitive methods but they are cumbersome and lengthy, even though simplifications have been proposed. Immunological techniques are easier, however they do not explore the biological activity of the circulating interferon. The presence of type I interferon in biological samples (serum, plasma, cerebro-spinal fluid, cultured cell supernatants) can be indirectly assessed by capability of interferon alpha to induce in vitro the synthesis of MxA in a dose dependent manner in cultured cells. Following to the lysis of the cells, the induced MxA can be quantitated and hence the type I-interferon concentration can be determinated in samples. The quantitation of MxA protein in peripheral blood lysates can be useful as a specific marker of acute viral infections. A minute amount of whole blood (15 mul) is sufficient which facilitates its use in pediatrics. The specifically type-I-interferons inducible MxA protein is also a potential useful marker in the management of interferon alpha-treatment. Moreover, the detection of interferon alpha and antiviral proteins constitute an indirect approach for investigating the hypothesis of the role of viruses in chronic diseases with suspected infectious aetiology.


Acute oral calcium-sodium citrate load in healthy males. Effects on acid-base and mineral metabolism, oxalate and other risk factors of stone formation in urine.
The currently preferred calcium preparations for supplementation of food vary widely with respect to calcium availability, effects on systemic mineral metabolism, acid-base status, and the calciuria-induced risk of urinary tract stone formation. In eight healthy males we studied the response to an acute load with alkali(sodium)-containing soluble calcium citrate (CSC) (molar ratio calcium/sodium/citrate approx. = 1/1/1), when taken in three different doses (10, 20, 30 mmol calcium) together with a continental breakfast. Intestinal calcium absorption, serum calcium, calcitonin, parathyroid hormone (PTH) other markers of bone metabolism, net acid excretion and calcium oxalate crystallization in urine were evaluated. CSC evoked a dose-dependent increase in calcium absorption, calcium in serum and urine, but no overt hypercalcemia, and calciuria was low relative to the excess calcium ingested; PTH fell and calcitonin rose (p < 0.05 vs. breakfast alone), but the diet-independent markers of bone resorption declined only insignificantly, while the markers of bone formation and turnover remained unchanged. There was a significant "once-daily" effect (= cumulative 24 h postload response) of CSC: a decrease in urinary cyclic AMP, phosphorus, and ammonium, and an increase in urinary bicarbonate. Soon after CSC intake, urinary calcium oxalate and hydroxyapatite supersaturation increased dose-dependently, the calcium oxalate crystal diameter was increased, but crystal aggregation time, which is crucial for stone formation, remained statistically unchanged. Thus, CSC provides calcium in a bioavailable form, creates mild systemic alkalinisation and inhibition of bone resorption, but leaves the risk of developing urinary stones unchanged. Comparative long-term studies on bone growth and the maintenance of bone health, using alkali-containing versus alkali-free calcium citrate, appear worthwhile.


Medicaid Contributes Substantial Costs to Dementia Care in an Ethnically Diverse Community.
The main objective of this study was to estimate effects of dementia on Medicaid expenditures in an ethnically diverse community. The sample included 1,211 Medicare beneficiaries who did not have any Medicaid coverage and 568 who additionally had full Medicaid coverage enrolled in the Washington Heights-Inwood Columbia Aging Project (WHICAP), a multiethnic, population-based, prospective study of cognitive aging in northern Manhattan (1999-2010). Individuals' dementia status was determined using a rigorous clinical protocol. Relationship between dementia and Medicaid coverage and expenditures were estimated using a two-part model. In participants who had full Medicaid coverage, average annual Medicaid expenditures were substantially higher for those with dementia than those without dementia ($50,270 vs. $21,966, p < .001), but Medicare expenditures did not differ by dementia status ($8,458 vs. $9,324, p = .19). In participants who did not have any Medicaid coverage, average annual Medicare expenditures were substantially higher for those with dementia than those without dementia ($12,408 vs. $8,113, p = .02). In adjusted models, dementia was associated with a $6,278 increase in annual Medicaid spending per person after controlling for other characteristics. Results highlight Medicaid's contribution to covering the cost of dementia care in addition to Medicare. Studies that do not include Medicaid are unlikely to accurately reflect the true cost of dementia.


Images depicting human pain increase exercise-induced pain and impair endurance cycling performance.
The current study investigated whether viewing images of others in pain influences exercise-induced pain (EIP) and cycling performance. Twenty-one recreational cyclists attended five laboratory visits. The first two visits involved measuring participants' maximal aerobic capacity and familiarized participants to the fixed power (FP) and 16.1 km cycling time trial (TT) tasks. The FP task required participants to cycle at 70% of their maximal aerobic power for 10-minutes. In the subsequent three visits, participants performed the FP and TT tasks after viewing pleasant, painful or neutral images. Participants rated the subset of painful images as more painful than the pleasant and neutral images; with no difference in the pain ratings of the pleasant and neutral images. In the FP task, EIP ratings were higher following painful compared to pleasant images, while no differences in EIP were observed between any other condition . In the TT, performance did not differ between the pleasant and neutral conditions. However, TT performance was reduced after viewing painful images compared to neutral or pleasant images. HR, B[La], perceived exertion and EIP did not differ between the three conditions. These results suggest that viewing painful images decreases TT performance and increases pain during fixed intensity cycling. Abbreviations: EIP: Exercise Induced Pain; FP: Fixed Power; TT: Time Trial; HR:Heart Rate; B[La]: Blood Lactate; RPE: Rating of Perceived Exertion; IAPS: International Affective Picture System; PO: Power Output.


Localization and Quantitative Autoradiography of Glutamatergic Ligand Binding Sites in Chick Brain.
The anatomical localization of glutamate receptor subtype-selective ligand binding sites was investigated in 1-day-old chick brain using quantitative autoradiography. Under the conditions used, the regional distributions of [3H]glutamate, [3H]AMPA (a selective quisqualate receptor ligand) and [3H]kainate binding sites are manifestly different. [3H]l-glutamate binding is densely localized in the telencephalon, particularly in the neostriatum (2.8 pmol/mg protein). In addition, [3H]l-glutamate labels the thalamus, the nucleus mesencephalicus lateralis pars dorsalis, the superficial layers of the optic tectum and the molecular layer of the cerebellum. [3H]AMPA binding sites are most densely localized in the hippocampus (0.90 pmol/mg protein), with an otherwise relatively uniform distribution of binding within the telencephalon. [3H]AMPA also labels the striatum griseum et fibrosum superficiale of the optic tectum and the molecular layer of the cerebellum. [3H]Kainate binding sites are extremely densely packed in the molecular layer of the cerebellum (10 pmol/mg protein). Other regions of [3H]kainate binding include the hyperstriatum and the thalamus. The binding of the NMDA receptor channel blocker [3H]MK-801 is increased in the presence of 1 mM l-glutamate. [3H]MK-801 binding is generally widespread in the telencephalon but is notably absent from the ectostriatum. No evidence of [3H]MK-801 binding sites was detected in the cerebellum, even in the presence of 1 mM l-glutamate. The relatively high densities and the well-defined localizations of the glutamate receptor subtype binding sites suggest that chick brain provides a useful system for the further study of excitatory amino acid receptors.


Measurements of hyperpolarized gas properties in the lung. Part III: (3)He T(1).
Hyperpolarized (3)He spin-lattice relaxation was investigated in the guinea pig lung using spectroscopy and imaging techniques with a repetitive RF pulse series. T(1) was dominated by interactions with oxygen and was used to measure the alveolar O(2) partial pressure. In animals ventilated with a mixture of 79% (3)He and 21% O(2), T(1) dropped from 19.6 sec in vivo to 14.6 sec after cardiac arrest, reflecting the termination of the intrapulmonary gas exchange. The initial difference in oxygen concentration between inspired and alveolar air, and the temporal decay during apnea were related to functional parameters. Estimates of oxygen uptake were 29 +/- 11 mL min(-1) kg(-1) under normoxic conditions, and 9.0 +/- 2.0 mL min(-1) kg(-1) under hypoxic conditions. Cardiac output was estimated to be 400 +/- 160 mL min(-1) kg(-1). The functional residual capacity derived from spirometric magnetic resonance experiments varied with body mass between 5.4 +/- 0.3 mL and 10.7 +/- 1.1 mL. Magn Reson Med 45:421-430, 2001.


Exopolysaccharide-deficient mutants of Rhizobium meliloti that form ineffective nodules.
By screening with the fluorescent stain Calcofluor, we have isolated 26 independent transposon Tn5 insertion mutants of Rhizobium meliloti that are deficient in the production of a known extracellular polysaccharide (Exo-). The mutants belonged to six distinct genetic groups based on the ability of their Exo- phenotype to be complemented by different recombinant plasmids from a R. meliloti clone bank. With few exceptions, all of the mutants formed ineffective (non-nitrogen-fixing) nodules on alfalfa. For all but one group, the complementing plasmids restored effective nodulation. These results establish a firm and extensive correlation between the ability of Rhizobium to produce a particular polysaccharide and symbiotic proficiency. The ineffective nodules appeared to contain no bacteroids and to form without shepherds' crooks or infection threads; this symbiotic phenotype matches that described for a set of independently isolated mutants that belong phenotypically and genetically to the group B exopolysaccharide mutants described previously [Finan et al. (1985) Cell 40, 869-877]. Apparently the exopolysaccharide, although not required for nodule formation, is involved in wild-type nodule invasion.


From clinical trials to clinical practice: an overview of Certican (everolimus) in heart transplantation.
Cardiac allograft vasculopathy (CAV) is the primary cause of late morbidity and mortality in heart transplant patients and remains a major challenge to further improvements in long-term graft survival in this population. Clearly, there is a need for immunosuppressive regimens that reduce the risk of CAV. Certican (everolimus) is a proliferation signal inhibitor developed for the prevention of acute and chronic rejection after solid-organ transplantation. Pre-clinical studies suggest that everolimus prevents vascular remodeling and neointimal proliferation, which are key components of CAV. In a pivotal trial in heart transplantation, everolimus at 1.5 or 3.0 mg plus standard-dose cyclosporine (CsA; Neoral) and corticosteroids demonstrated superior efficacy to azathioprine (AZA) by decreasing the incidence of biopsy-proven acute rejection (BPAR) and the composite end-point, efficacy failure. Importantly, in this trial, everolimus was also associated with a significant reduction in both the incidence and severity of CAV in recipients of heart transplants. Furthermore, cytomegalovirus (CMV) infection rates were significantly lower with everolimus than with AZA. The study suggests that everolimus has the ability to target the primary causes of chronic allograft dysfunction by reducing acute rejection and CMV infection, and preventing CAV. Moreover, these findings indicate that use of everolimus as part of the primary immunosuppression regimen, could provide a major benefit for heart transplant patients, offering a real hope of alleviating CAV in the long term. Few large-scale trials have been conducted in heart transplant patients, so their value must therefore be maximized with findings being effectively translated into clinical practice.


Diseases of poverty with high mortality in infants and children: malaria, measles, lower respiratory infections, and diarrheal illnesses.
Malaria, measles, lower respiratory infections, and diarrheal illnesses are common pediatric medical problems that are often fatal in the context of extreme poverty. In nonpoor environments, however, these infections are controlled and managed in ways that minimize mortality. From a scientific perspective, genetic variation among microbes is a frequent and important component of their epidemiology, pathophysiology, treatment, and prevention. From a public health perspective, relatively simple measures can reduce the mortal effects of these diseases until successful vaccines become available and immunizations programs are established. Infants and children are especially vulnerable to poor outcomes from infections when undernutrition and other circumstances of poverty are present.


Progress in primary aldosteronism: mineralocorticoid receptor antagonists and management of primary aldosteronism in pregnancy.
Primary aldosteronism (PA) is the most common cause of secondary hypertension. In this review, we discuss the diagnosis and management of PA during pregnancy based on the literature. As aldosterone and renin are physiologically increased during pregnancy and confirmation tests are not recommended, the diagnosis of PA during pregnancy relies on a repeatedly suppressed plasma renin level. Mineralocorticoid receptor antagonists (MRAs) are the most effective drugs to treat hypertension and hypokalemia in patients with PA. However, spironolactone (FDA pregnancy category C) might lead to undervirilization of male infants due to the anti-androgenic effects. Although data in the literature are very limited, treatment with spironolactone is not recommended. Eplerenone (FDA pregnancy category B) is a selective MRA without anti-androgenic potential. If MRA treatment is required in pregnancy, eplerenone appears to be a safe and effective alternative, although symptomatic treatment with approved antihypertensive drugs and supplementation with potassium is the first choice. In case of aldosterone-producing adenoma, laparoscopic adrenalectomy is a therapeutic option in the second trimester of pregnancy.


Systemic Insecticide Applications: Effects on Citrus Mealybug (Hemiptera: Pseudococcidae) Populations Under Greenhouse Conditions.
The citrus mealybug, Planococcus citri (Risso) (Hemiptera: Pseudococcidae), is a major insect pest of greenhouse-grown horticultural crops. Applications of systemic insecticides as drenches to the growing medium are typically used by greenhouse producers to prevent or suppress citrus mealybug populations. A comprehensive study was conducted that included 11 experimental trials designed to assess the effects of growing medium applications of six systemic insecticides (azadirachtin, dinotefuran, flonicamid, imidacloprid, spirotetramat, and thiamethoxam) in preventing the establishment of and suppressing citrus mealybug populations on different plant species. The experimental trials included four plant species: Solenostemon scutellarioides, Gerbera jamesonii, Begonia × semperflorens-cultorum, and Salvia splendens, with six different cultivars grown in research greenhouses similar to those used in greenhouse production systems. In addition, feeding location (plant stem, leaf top, and leaf bottom) of citrus mealybugs on the various plants was also recorded. Overall, percent citrus mealybug mortality was consistently <50% (n = 255 to 1,598) for any treatment or rate of application; regardless if the systemic insecticides were applied preventatively or curatively. Percent citrus mealybug mortality did reach 78% (n = 36) for thiamethoxam at 8 times the label rate when plants were treated preventatively. Furthermore, none of the treatments prevented development of citrus mealybug F1 individuals to F2 generation egg-laying females on S. scutellarioides or G. jamesonii plants. Citrus mealybugs varied in their spatial distribution on the plant stem, leaf top, and leaf bottom with no noticeable trends. Therefore, based on the results of the study, systemic insecticides are not effective against the citrus mealybug in greenhouse production systems.


CWPO of 4-CP and industrial wastewater with Al-Fe pillared clays.
Catalysts based on pillared clays with Al-Fe have been synthesised from a commercial bentonite and tested for catalytic wet peroxide oxidation (CWPO) of aqueous 4-Chlorophenol (4-CP) solution and industrial wastewater from cosmetics manufacture. The effect of the synthesis procedure, the iron load and reaction temperature on the catalytic activity was studied using 4-CP as target compound. A lower temperature in the preparation of the pillaring solution, as well as a higher Fe load, gave rise to a higher catalytic activity, but also a higher leaching of the active phase. The best catalyst, in terms of catalytic activity, was also tested for treating cosmetic wastewater by CWPO. Experiments were carried out at 90 degrees C and atmospheric pressure and the influence of Fe load, catalyst concentration and H(2)O(2)/COD ratio (between 0.5 and 2 times the stoichiometric ratio) were analysed. Higher values of these parameters favour COD reduction. The Fe leaching in all cases was lower than 1.2 mg/L, indicating that these catalysts have a high stability under these experimental conditions.


Studies of JC virus-induced nervous system tumors in the Syrian hamster: a review.
A review has been presented of the essential primary tumors of the nervous system, and of one secondary tumor of the brain, which has been obtained after JC virus inoculation of Syrian hamsters. These results have been compared with results obtained by other authors, testing different group B papovaviruses. Comparisons have also been made with neurooncogenicity results obtained with human adenovirus 12 in rodents and with JC virus in subhuman primates. The similarity of the phenotypically diverse JC virus-induced hamster tumors with human childhood of the nervous system and their usefulness for extended biologic research has been mentioned.


Adding learning to cellular genetic algorithms for training recurrent neural networks.
This paper proposes a hybrid optimization algorithm which combines the efforts of local search (individual learning) and cellular genetic algorithms (GA's) for training recurrent neural networks (RNN's). Each weight of an RNN is encoded as a floating point number, and a concatenation of the numbers forms a chromosome. Reproduction takes place locally in a square grid with each grid point representing a chromosome. Two approaches, Lamarckian and Baldwinian mechanisms, for combining cellular GA's and learning have been compared. Different hill-climbing algorithms are incorporated into the cellular GA's as learning methods. These include the real-time recurrent learning (RTRL) and its simplified versions, and the delta rule. The RTRL algorithm has been successively simplified by freezing some of the weights to form simplified versions. The delta rule, which is the simplest form of learning, has been implemented by considering the RNN's as feedforward networks during learning. The hybrid algorithms are used to train the RNN's to solve a long-term dependency problem. The results show that Baldwinian learning is inefficient in assisting the cellular GA. It is conjectured that the more difficult it is for genetic operations to produce the genotypic changes that match the phenotypic changes due to learning, the poorer is the convergence of Baldwinian learning. Most of the combinations using the Lamarckian mechanism show an improvement in reducing the number of generations required for an optimum network; however, only a few can reduce the actual time taken. Embedding the delta rule in the cellular GA's has been found to be the fastest method. It is also concluded that learning should not be too extensive if the hybrid algorithm is to be benefit from learning.


Noninvasive biomarkers in non-alcoholic fatty liver disease: current status and a glimpse of the future.
The development of non invasive biomarkers of disease has become a major focus of interest in nonalcoholic fatty liver disease (NAFLD). The large prevalence of the disease and the invasive nature of the investigation means that screening with liver biopsy is impractical. In addition to screening, the differentiation of those with simple steatosis vs steatohepatitis and fibrosis is clinically important as the prognosis of each differs. Serum biomarkers may be a combination of simple markers derived from large data sets or direct markers of disease activity. Serum markers of inflammation, apoptosis and oxidative stress in addition to fibrosis have been extensively studied in patients with NAFLD. Other techniques such as transient elastography, magnetic resonance elastography and acoustic radiation force imaging are becoming more established as noninvasive methods of detecting fibrosis in a variety of chronic liver conditions in addition to NAFLD. Newer high throughput methods such as proteomics and glycomics allow the nonhypothesis-driven identification of novel markers and may also potentially contribute to our understanding of the pathogenesis of the condition. This review addresses some of the methodological issues which need to be considered in the search for the ideal biomarker. It is likely that a combination of serum biomarkers and techniques such as transient elastography may provide the optimal diagnostic discrimination however this remains to be proven in large studies.


The three isoforms of the light-harvesting complex II: spectroscopic features, trimer formation, and functional roles.
The major light-harvesting complex (LHC-II) of higher plants plays a crucial role in capturing light energy for photosynthesis and in regulating the flow of energy within the photosynthetic apparatus. Native LHC-II isolated from plant tissue consists of three isoforms, Lhcb1, Lhcb2, and Lhcb3, which form homo- and heterotrimers. All three isoforms are highly conserved among different species, suggesting distinct functional roles. We produced the three LHC-II isoforms by heterologous expression of the polypeptide in Escherichia coli and in vitro refolding with purified pigments. Although Lhcb1 and Lhcb2 are very similar in polypeptide sequence and pigment content, Lhcb3 is clearly different because it lacks an N-terminal phosphorylation site and has a higher chlorophyll a/b ratio, suggesting the absence of one chlorophyll b. Low temperature absorption and fluorescence emission spectra of the pure isoforms revealed small but significant differences in pigment organization. The oligomeric state of the pure isoforms and of their permutations was investigated by native gel electrophoresis, sucrose density gradient centrifugation, and SDS-PAGE. Lhcb1 and Lhcb2 formed trimeric complexes by themselves and with one another, but Lhcb3 was able to do so only in combination with one or both of the other isoforms. We conclude that the main role of Lhcb1 and Lhcb2 is in the adaptation of photosynthesis to different light regimes. The most likely role of Lhcb3 is as an intermediary in light energy transfer from the main Lhcb1/Lhcb2 antenna to the photosystem II core.


Cytotoxic Pathways in Allogeneic Hematopoietic Cell Transplantation.
Allogeneic hematopoietic cell transplantation (allo-HCT) is a potentially curative treatment for hematologic malignancies, and other hematologic and immunologic diseases. Donor-derived immune cells identify and attack cancer cells in the patient producing a unique graft-vs.-tumor (GVT) effect. This beneficial response renders allo-HCT one of the most effective forms of tumor immunotherapy. However, alloreactive donor T cells can damage normal host cells thereby causing graft-vs.-host disease (GVHD), which results in substantial morbidity and mortality. To date, GVHD remains as the major obstacle for more successful application of allo-HCT. Of special significance in this context are a number of cytotoxic pathways that are involved in GVHD and GVT response as well as donor cell engraftment. In this review, we summarize progress in the investigation of these cytotoxic pathways, including Fas/Fas ligand (FasL), perforin/granzyme, and cytokine pathways. Many studies have delineated their distinct operating mechanisms and how they are involved in the complex cellular interactions amongst donor, host, tumor, and infectious pathogens. Driven by progressing elucidation of their contributions in immune reconstitution and regulation, various interventional strategies targeting these pathways have entered translational stages with aims to improve the effectiveness of allo-HCT.


Psychiatric Medication Intake in Suicide Victims: Gender Disparities and Implications for Suicide Prevention.
Frequency and gender differences of psychiatric medication intake in a sample of suicide victims from the Athens Greater Area were investigated with a particular focus on the implications for suicide prevention. Data were collected from the toxicological analyses of the suicide cases of the period November 2007-October 2009. Information was available for 262 individuals, 196 men (74.8%) and 66 women (25.2%); 109 of these (41.6%) were receiving psychiatric medication(s). Women were statistically more frequently under treatment: antidepressants (32.8% vs. 11.3%, p < 0.001), antiepileptics (9.1% vs. 0.5%, p = 0.001), antipsychotics (24.2% vs. 9.2%, p = 0.003), and benzodiazepines (16.7% vs. 6.6%, p = 0.024). Campaigns aiming to bring men with psychological difficulties in contact with mental health services and to lessen the stigma of mental illness, together with better training of nonpsychiatrists into "suspecting" "male" depression, could be particularly helpful for decreasing male suicides. More thoughtful choice of psychiatric medication could possibly already prevent a number of female suicides.


cGMP signaling in vertebrate retinal photoreceptor cells.
The visual transduction pathway in vertebrate photoreceptors transforms a light stimulus entering the photoreceptor outer segments into an electrical response at the synapses of rod and cone photoreceptor cells. This process is mediated by complex biochemical pathways that precisely regulate cGMP levels, thereby controlling the extent, duration, and adaptation of the photoreceptor to the light stimulus. This review first summarizes the major mechanisms of regulating cytoplasmic cGMP levels (synthesis, degradation, buffering, and efflux) as well as the primary targets of action of cGMP (cyclic nucleotide-gated ion channels, cGMP-dependent protein kinase, and cGMPregulated phosphodiesterases). This information is applied to our current understanding of how these processes operate in the signal-transducing outer segment of rod and cone photoreceptors to carry out visual excitation, recovery, and adaptation in response to light stimulation.


Preoperative resuscitation of the trauma patient.
The traditional approach to trauma patients with presumed internal hemorrhage has been immediate, aggressive intravenous fluid resuscitation. Recent experimental and clinical data, however, suggest a more discriminating approach that first considers concurrent head injury, hemodynamic stability and the presence of potentially uncontrollable hemorrhage (e.g. deep truncal injury) versus a controllable source (e.g. distal extremity wound) as well as the use of new techniques to inhibit bleeding and better ways to monitor the patient's condition. Evolving data suggest that while aggressive fluid infusions could be used for patients with isolated extremity, thermal or head injury, they should be limited in conditions with potentially uncontrollable internal hemorrhage, and particularly in patients with penetrating truncal injury being transported immediately to a trauma center. Likewise, the minute volume of positive pressure ventilatory support should be limited with potential severe hemorrhage due to the secondary reductions in venous return. For trauma patients with severe bleeding there is growing evidence for the increased use of plasma and factor VIIa, as well as tourniquets, intraosseus devices and evolving monitoring techniques. Owing to the growing societal threat of trauma, further research, including studies already under way, will be critical to delineate the timing and technique of infusing advantageous resuscitative fluids such as hypertonic saline and hemoglobin-based oxygen carriers as well as the use of hemostatic agents and special blood products.


Mirroring activity in the brain and movement determinant in the Rorschach test.
Human movement (M) responses to the Rorschach are related to cognitive sophistication, creativity, and empathy. Recent studies also link Ms to EEG-mu suppression, an index of mirroring activity in the brain. In this article, we further investigate the link between Ms and mu suppression by testing some clinical interpretative distinctions. Previously collected EEG data recorded during the administration of the Rorschach were reanalyzed. We hypothesized that (a) among several responses investigated, only M would be associated with mu suppression, and (b) Ms with active movement, ordinary form quality, or whole human figures would be most strongly associated with mu suppression. Hypothesis 1 was fully confirmed, thus supporting that the traditional interpretation of M has a neurobiological foundation. Hypothesis 2 was partially confirmed; that is, active Ms were associated with mu suppression more strongly than passive Ms (p < .05), but no other significant differences emerged. Clinical implications are discussed.


Replication stress, DNA damage signalling, and cytomegalovirus infection in human medulloblastomas.
Medulloblastomas are the most common, and often fatal, paediatric brain tumours that feature high genomic instability, frequent infection by human cytomegalovirus (HCMV) and resistance to radiation and chemotherapy. The causes of the pronounced chromosomal instability and its potential links with HCMV infection and/or resistance to genotoxic therapies remain largely unknown. To address these issues, here we have combined immunohistochemical analysis of a series of 25 paediatric medulloblastomas, complemented by medulloblastoma cell culture models including experimental HCMV infection. Using eight established immunohistochemical markers to assess the status of the DDR machinery, we found pronounced endogenous DNA damage signalling (γH2AX marker) and robust constitutive activation of both the ATM-Chk2 and ATR-Chk1 DNA damage checkpoint kinase cascades, yet unexpectedly modest p53 tumour suppressor activation, across our medulloblastoma cohort. Most tumours showed high proliferation (Ki67 marker), variable oxidative DNA damage (8-oxoguanine lesions) and formation of 53BP1 nuclear 'bodies', the latter indicating (along with ATR-Chk1 signalling) endogenous replication stress. The bulk of the clinical specimens also showed expression of HCMV immediate early and late proteins, in comparative analyses using three immunohistochemical protocols. Cell culture experiments validated the chronic endogenous replication stress in medulloblastoma cell lines and showed sharply differential, intriguing responses of normal cells and medulloblastoma cells to HCMV infection, including differential subcellular mislocalization and enhancement of replication stress-related 53BP1 body formation, the latter in cell-non-autonomous manner. Overall, our results strongly indicate that in human medulloblastomas, the DDR checkpoint barrier is widely activated, at least in part due to replication stress. Furthermore, we propose that unorthodox p53 defects other than mutations may allow high proliferation despite the ongoing checkpoint signalling and that the highly prevalent HCMV may impact the medulloblastoma host cell replication stress and DNA repair. Collectively, the scenario we report here likely fuels genomic instability and evolution of medulloblastoma resistance to standard-of-care genotoxic treatments.


Evaluation of bendectin embryotoxicity in nonhuman primates: I. Ventricular septal defects in prenatal macaques and baboon.
Cynomolgus monkeys, rhesus monkeys and baboons were administered 10 to 40 times the human dose equivalent of Bendectin throughout the major period of organogenesis (22(+/-3)-50 days of gestation). In animals examined prenatally (100 +/- 2 days gestation) the total incidence of ventricular septal defects (VSD) was 40% in cynomolgus monkeys, 18% in rhesus monkeys, and 23% in baboons. The majority of VSD involved the muscular portion of the septum. No dose response was evident and there were no other cardiac or extracardiac defects found except for one baboon fetus with multiple defects. No defects were observed in cynomolgus monkeys administered Bendectin for 4-day periods between 22 and 41 days of gestation. There was no association of Bendectin treatment with any noncardiac defect. In cynomolgus and rhesus monkeys examined at term there was one mitral valve defect and no incidence of VSD. The increased incidence of VSD observed prenatally in all three species and the absence of defects in macaques at term suggests a delay in closure of the ventricular septum in treated animals. The Bendectin-treated monkey may be a suitable model for the study of the pathogenesis of VSD and the mechanism of spontaneous closure of the defect.