Commentary: thirty years of AIDS in America: a story of infinite hope.
The year 2011 marks the thirtieth anniversary of the first case reports in the United States of what we now know to be end-stage HIV disease. This chronological milestone provides an opportunity to reflect upon the changing context of the American HIV/AIDS epidemic. Using two seminal documents as a framework, the 1986 Institute of Medicine Report, "Confronting AIDS," and the 2010 National HIV/AIDS Strategy, this descriptive analysis details our accomplishments in addressing the domestic U.S. epidemic and outlines what remains to be done on the long road to eradication of HIV disease. The past three decades have witnessed tremendous biomedical and behavioral advances in preventing, diagnosing, and treating HIV disease. However, to fully realize the promise of these scientific advances, such that we achieve the vision of the National HIV/AIDS Strategy, we must develop effective strategies to surmount a number of salient challenges, including: unbalanced combinations of prevention interventions; programs that are not of adequate scale to achieve population-level results; systems of service delivery that do not function in an integrated fashion; and social and economic structures that increase the vulnerability of populations who are at risk for or living with HIV disease.


Cimetidine suspension in patients with stage 0 gastro-oesophageal reflux disease.
One hundred and twenty-five patients with symptoms of heartburn and acid regurgitation but without endoscopic abnormalities were randomized to receive 200 mg cimetidine suspension four times daily or placebo for two weeks. Daily dairy cards were kept to evaluate the frequency and degree of symptoms. At two weeks cimetidine was significantly more effective than placebo. It is concluded that placebo suspension has a considerable effect on gastro-oesophageal reflux disease symptoms, but cimetidine suspension provides significantly better relief.


Sustained delayed gastric emptying during repeated restraint stress in oxytocin knockout mice.
We have recently shown that impaired gastric motility observed in acute restraint stress was restored following repeated restraint stress in mice. Repeated restraint stress up-regulates oxytocin mRNA expression and down-regulates corticotrophin-releasing factor (CRF) mRNA expression at the hypothalamus. Oxytocin knockout mice (OXT-KO) have been widely used to study the central oxytocin signalling pathways in response to various stressors. We studied the effects of acute and repeated restraint stress on solid gastric emptying and hypothalamic CRF mRNA expression in wild-type (WT) and OXT-KO mice. Heterozygous (HZ) parents (B6; 129S-Oxt(tm1Wsy)/J mice) were bred in our animal facility. Male OXT-KO, WT and HZ littermates were used for the study. Solid gastric emptying was measured following acute restraint stress (for 90 min) or repeated restraint stress (for five consecutive days). Expression of CRF mRNA in the paraventricular nucleus (PVN) was measured by real-time reverse transcriptase-polymerase chain reaction. There were no significant differences of gastric emptying in WT (68.4 ± 4.1%, n = 6), HZ (71.8 ± 3.1%, n = 6) and OXT-KO (70.6 ± 3.1%, n = 6) mice in nonstressed conditions. Acute stress significantly delayed gastric emptying in OXT-KO mice (33.10 ± 2.5%, n = 6) WT (39.1 ± 1.1%, n = 6) and HZ mice (35.8 ± 1.2%, n = 6). Following repeated restraint stress loading, gastric emptying was significantly restored in WT (68.3 ± 4.5%, n = 6) and HZ mice (63.1 ± 2.6%, n = 6). By contrast, gastric emptying was still delayed in OXT-KO mice (34.7 ± 1.3%, n = 6) following repeated restraint stress. The increase in CRF mRNA expression at the PVN was much pronounced in OXT-KO mice compared to WT or HZ mice following repeated restraint stress. These findings suggest that central oxytocin plays a pivotal role in mediating the adaptation mechanism following repeated restraint stress in mice.


Stereotypical cell division orientation controls neural rod midline formation in zebrafish.
The development of multicellular organisms is dependent on the tight coordination between tissue growth and morphogenesis. The stereotypical orientation of cell divisions has been proposed to be a fundamental mechanism by which proliferating and growing tissues take shape. However, the actual contribution of stereotypical division orientation (SDO) to tissue morphogenesis is unclear. In zebrafish, cell divisions with stereotypical orientation have been implicated in both body-axis elongation and neural rod formation, although there is little direct evidence for a critical function of SDO in either of these processes. Here we show that SDO is required for formation of the neural rod midline during neurulation but dispensable for elongation of the body axis during gastrulation. Our data indicate that SDO during both gastrulation and neurulation is dependent on the noncanonical Wnt receptor Frizzled 7 (Fz7) and that interfering with cell division orientation leads to severe defects in neural rod midline formation but not body-axis elongation. These findings suggest a novel function for Fz7-controlled cell division orientation in neural rod midline formation during neurulation.


Duration of Oxaliplatin-Containing Adjuvant Therapy for Stage III Colon Cancer: ASCO Clinical Practice Guideline.
To develop recommendations for duration of adjuvant chemotherapy with a fluoropyrimidine and oxaliplatin for patients with completely resected stage III colon cancer based on the results of trials of 3 months compared with 6 months of treatment. ASCO convened an Expert Panel and conducted a systematic review of relevant studies. The guideline recommendations were based on the review of evidence by the Expert Panel. Pooled data from the six International Duration Evaluation of Adjuvant Chemotherapy (IDEA) Collaboration randomized controlled trials comprise the evidence base for these guideline recommendations. The recommendations for therapy duration apply to patients with completely resected stage III colon cancer who are being offered adjuvant chemotherapy with oxaliplatin and a fluoropyrimidine. Recommendations are informed by the findings of a recent pooled analysis of clinical trials that compared 6 months versus 3 months of oxaliplatin-based chemotherapy. For patients at a high risk of recurrence (T4 and/or N2), adjuvant chemotherapy should be offered for a duration of 6 months. For patients at a low risk of recurrence (T1, T2, or T3 and N1), either 6 months of adjuvant chemotherapy or a shorter duration of 3 months may be offered on the basis of a potential reduction in adverse events and no significant difference in disease-free survival with the 3-month regimen. In determining duration of therapy, the Expert Panel recommends a shared decision-making approach, taking into account patient characteristics, values and preferences, and other factors and including a discussion of the potential for benefit and risks of harm associated with treatment duration. Additional information is available at www.asco.org/gastrointestinal-cancer-guidelines .


Trans-acting small RNA determines dominance relationships in Brassica self-incompatibility.
A diploid organism has two copies of each gene, one inherited from each parent. The expression of two inherited alleles is sometimes biased by the effects known as dominant/recessive relationships, which determine the final phenotype of the organism. To explore the mechanisms underlying these relationships, we have examined the monoallelic expression of S-locus protein 11 genes (SP11), which encode the male determinants of self-incompatibility in Brassica. We previously reported that SP11 expression was monoallelic in some S heterozygotes, and that the promoter regions of recessive SP11 alleles were specifically methylated in the anther tapetum. Here we show that this methylation is controlled by trans-acting small non-coding RNA (sRNA). We identified inverted genomic sequences that were similar to the recessive SP11 promoters in the flanking regions of dominant SP11 alleles. These sequences were specifically expressed in the anther tapetum and processed into 24-nucleotide sRNA, named SP11 methylation inducer (Smi). Introduction of the Smi genomic region into the recessive S homozygotes triggered the methylation of the promoter of recessive SP11 alleles and repressed their transcription. This is an example showing sRNA encoded in the flanking region of a dominant allele acts in trans to induce transcriptional silencing of the recessive allele. Our finding may provide new insights into the widespread monoallelic gene expression systems.


User-Centered System Design for Communicating Clinical Laboratory Test Results: Design and Evaluation Study.
Personal clinical data, such as laboratory test results, are increasingly being made available to patients via patient portals. However, laboratory test results are presented in a way that is difficult for patients to interpret and use. Furthermore, the indications of laboratory test results may vary among patients with different characteristics and from different medical contexts. To date, little is known about how to design patient-centered technology to facilitate the interpretation of laboratory test results. The aim of this study is to explore design considerations for supporting patient-centered communication and comprehension of laboratory test results, as well as discussions between patients and health care providers. We conducted a user-centered, multicomponent design research consisting of user studies, an iterative prototype design, and pilot user evaluations, to explore design concepts and considerations that are useful for supporting patients in not only viewing but also interpreting and acting upon laboratory test results. The user study results informed the iterative design of a system prototype, which had several interactive features: using graphical representations and clear takeaway messages to convey the concerning nature of the results; enabling users to annotate laboratory test reports; clarifying medical jargon using nontechnical verbiage and allowing users to interact with the medical terms (eg, saving, favoriting, or sorting); and providing pertinent and reliable information to help patients comprehend test results within their medical context. The results of a pilot user evaluation with 8 patients showed that the new patient-facing system was perceived as useful in not only presenting laboratory test results to patients in a meaningful way but also facilitating in situ patient-provider interactions. We draw on our findings to discuss design implications for supporting patient-centered communication of laboratory test results and how to make technology support informative, trustworthy, and empathetic.


Comparison of the effectiveness of endotracheal tube holder with the conventional method in a manikin model.
Endotracheal tube (ETT) displacement occurs by improper fixation. To fix an ETT, many types of fixation tools are employed. Thomas tube holder is one of the fixation tools widely used in many countries. This study aims to compare the ETT fixation using the Thomas tube holder with the conventional method (adhesive tape) in a mannequin model. The fixation tools were random, using the box of six randomizes to Thomas tube holder and conventional method. After fixation, the mannequin model was being logged roll, chest compression by automated chest compression machine, and transported by the paramedic. The time to ETT fixation and displacements were recorded. The mean time (standard deviation) to fixate an ETT was shorter (33.0 s [7.3]) with a Thomas tube holder compared to adhesive tape (52.6 s [7.3], P < 0.001). The number and proportion of the ETT displacements were significantly less with Thomas tube holder compared to adhesive tape during log roll (16, 35.6% vs. 29, 64.4%, P = 0.011), chest compression with automated machine (23, 51.1% vs. 37, 82.2%, P = 0.003), and transport (26, 57.8% vs. 40, 88.9%, P = 0.002). The Thomas tube holder is more effective than adhesive tape in preventing ETT displacement in a mannequin subjected to log roll, chest compressions, and transportation.


From Favorable Histology to Relapse: The Clonal Evolution of a Wilms Tumor.
Favorable histology (FH) Wilms tumor (WT) is one of the most curable of all human cancers, yet a small minority of patients fail treatment. The underlying biological pathways that lead to therapy resistance are unknown. We report a case of initially unresectable, FH WT which revealed limited necrosis and persistent blastemal predominant histology following neoadjuvant chemotherapy. Despite intensification of therapy and whole abdominal radiation, the patient relapsed and succumbed to her disease. In an effort to discover candidate drivers of drug resistance, whole exome sequencing and copy number analysis were performed on samples from all 3 tumor specimens. Sequencing results revealed outgrowth of clones with a dramatically different genetic landscape including dominant mutations that could explain therapy evasion, some of which have not been previously reported in WT. Our results implicate PPM1D, previously shown to be associated with drug resistance in other tumors, as the major driver of treatment failure.


[Effects of hypothermal and circulatory arrest on the auditory brainstem response during operation in children].
Twenty-two children were monitored by the auditory brainstem response (ABR) during the open chest and/or open heart surgery for cardiac anomalies under the extracorporeal circulation (ECC) with moderate to deep hypothermia. These patients were divided into two groups, namely, the ECC only group (group A: 9 cases) and the total circulatory arrest (TCA) group (group B: 13 cases). The mean age of group A was 36 months and group B was 6.1 months. In group A, moderate hypothermia was conducted with the rectal temperature ranging from 25 degrees C to 30 degrees C, and in group B profound hypothermia was conducted lowering the temperature to 18 degrees C. The ABR recorded at the following points, namely; before inducing anesthesia, before lowering the body temperatures, during the cooling process, at the time of TCA, and during the rewarming process. Accompanying the decrease in body temperature the peak latency of waves I, III and V were markedly prolonged. When the rectal temperature fell to 23 degrees C, the peak latency of each wave was prolonged to about 150% of their precooling values at 36 degrees C. When it fell to 22 degrees C, the ABR disappeared entirely in 16 of 22 cases, and in remaining 6 cases, only I wave was detected. During rewarming, in both the A and B groups, at 24 degrees C, the wave of ABR started to reappear beginning with I waves, and on reaching 26 degrees C, I, III and V waves from became detectable. The peak latency of all waves at rectal temperature of 33 degrees C recovered to almost the same values as these at 36 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)


Low threshold calcium currents in rat cerebellar Purkinje cell dendritic spines are mediated by T-type calcium channels.
The functional role of low voltage activated (LVA) calcium channels in the cerebellar Purkinje cell dendritic tree is not completely understood. Since the localization of these channels will influence their possible roles in dendritic integration and induction of plasticity, we set out to characterize the LVA calcium current in Purkinje cell dendrites in acute cerebellar slices of young rats. Using a combination of electrophysiological recordings and two-photon laser scanning microscopy, we show that LVA calcium current recorded at the soma can be correlated with voltage-dependent calcium transients in Purkinje cell dendritic spines. Blocking sodium and potassium conductances allowed us to isolate and characterize a fast inactivating inward current activated positive to -55 mV. Activation and steady-state inactivation kinetics, voltage-dependent deactivation kinetics, and pharmacological experiments (using omega-agatoxin-IVA, mibefradil and nickel) show that this current is carried by T-type calcium channels. Furthermore, the LVA calcium transient observed in the dendritic spines of the Purkinje cell is well correlated with the current recorded at the soma, suggesting that T-type calcium channels are the main component of the LVA calcium input in spines. The fast rising phase of the calcium transient in spines and the absence of delay between the onset in the spine and the parent dendrite show that T-type calcium channels are present both in spines and dendrites of the Purkinje cell.


Effect of maximal voluntary contraction on the amplitude of the compound muscle action potential: implications for the interpolated twitch technique.
The compound muscle action potential (M(MAX)) during a maximal voluntary contraction (MVC) may be measured to determine if the motor nerve has been supramaximally stimulated during the interpolated twitch technique (ITT). Ten males performed isometric knee extension MVCs. M(MAX) for the vastus medialis was recorded during MVC and rest. To examine the effect of stimulating electrode movement, the M(MAX) of the thenar group and antidromic sensory nerve action potentials (SNAPs) to the third digit were recorded in a separate experiment. M(MAX) during MVC was reduced by 18% (P < 0.0001) and 43% (p < 0.0001) for the quadriceps and thenar group, respectively. The SNAP amplitude was not different between rest and MVC (P = 0.18). Reduction of M(MAX) during MVC suggests that some motor axons are refractory and unable to respond to a superimposed maximal stimulus. These results have implications for the sensitivity of the interpolated twitch technique.


The changing balance of brainstem-spinal cord modulation of pain processing over the first weeks of rat postnatal life.
Brainstem-spinal cord connections play an essential role in adult pain processing, and the modulation of spinal pain network excitability by brainstem nuclei is known to contribute to hyperalgesia and chronic pain. Less well understood is the role of descending brainstem pathways in young animals when pain networks are more excitable and exposure to injury and stress can lead to permanent modulation of pain processing. Here we show that up to postnatal day 21 (P21) in the rat, the rostroventral medulla of the brainstem (RVM) exclusively facilitates spinal pain transmission but that after this age (P28 to adult), the influence of the RVM shifts to biphasic facilitation and inhibition. Graded electrical microstimulation of the RVM at different postnatal ages revealed a robust shift in the balance of descending control of both spinal nociceptive flexion reflex EMG activity and individual dorsal horn neuron firing properties, from excitation to inhibition, beginning after P21. The shift in polarity of descending control was also observed following excitotoxic lesions of the RVM in adult and P21 rats. In adults, RVM lesions decreased behavioural mechanical sensory reflex thresholds, whereas the same lesion in P21 rats increased thresholds. These data demonstrate, for the first time, the changing postnatal influence of the RVM in spinal nociception and highlight the central role of descending brainstem control in the maturation of pain processing.


Alteration in RGS2 expression level is associated with changes in haloperidol induced extrapyramidal features in a mutant mouse model.
Antipsychotic induced Parkinsonism (AIP) is a common adverse effect of antipsychotic drug treatment among schizophrenia patients. Two previous studies showed association of the rs4606 SNP in the 3' untranslated region of the regulator of G protein signaling 2 gene (RGS2) with susceptibility to AIP. Since rs4606 reportedly influences expression of RGS2, we applied a translational approach and studied the effect of chronic (24 days) exposure to haloperidol on AIP-like features in mice carrying a mutation that causes lower Rgs2 gene expression. Haloperidol and vehicle treated male mice heterozygous (HET) or homozygous (HOM) for the mutation, or wild type (WT), were evaluated for open field locomotion, catalepsy duration, pole test performance and rota-rod latency to fall. We showed that in haloperidol treated mice lower Rgs2 expression is associated with better performance on the open field, catalepsy and rota-rod tests but not the pole test. Results were most consistent for the 0.2 mg/kg/d haloperidol dose. These observations support the possible involvement of RGS2 in mechanisms underlying susceptibility to AIP.


Modulation of ileal apical Na+-dependent bile acid transporter ASBT by protein kinase C.
Ileal apical Na(+)-dependent bile acid transporter (ASBT) is responsible for reabsorbing the majority of bile acids from the intestinal lumen. Rapid adaptation of ASBT function in response to physiological and pathophysiological stimuli is essential for the maintenance of bile acid homeostasis. However, not much is known about molecular mechanisms responsible for acute posttranscriptional regulation of ileal ASBT. The protein kinase C (PKC)-dependent pathway represents a major cell signaling mechanism influencing intestinal epithelial functions. The present studies were, therefore, undertaken to investigate ASBT regulation in intestinal Caco-2 monolayers by the well-known PKC activator phorbol 12-myristate 13-acetate (PMA). Our results showed that Na(+)-dependent [(3)H]taurocholic acid uptake in Caco-2 cells was significantly inhibited in response to 2 h incubation with 100 nM PMA compared with incubation with 4alpha-PMA (inactive form). The inhibitory effect of PMA was blocked in the presence of 5 microM bisindolylmaleimide I (PKC inhibitor) but not 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-AM (Ca(2+) chelator) or LY-294002 (phosphatidylinositol 3-kinase inhibitor). PMA inhibition of ASBT function was also abrogated in the presence of myristoylated PKCzeta pseudosubstrate peptide, indicating involvement of the atypical PKCzeta isoform. The inhibition by PMA was associated with a significant decrease in the maximal velocity of the transporter and a reduction in ASBT plasma membrane content, suggesting a modulation by vesicular recycling. Our novel findings demonstrate a posttranscriptional modulation of ileal ASBT function and membrane expression by phorbol ester via a PKCzeta-dependent pathway.


Monitoring Needs for Gene Drive Mosquito Projects: Lessons From Vector Control Field Trials and Invasive Species.
As gene drive mosquito projects advance from contained laboratory testing to semi-field testing and small-scale field trials, there is a need to assess monitoring requirements to: i) assist with the effective introduction of the gene drive system at field sites, and ii) detect unintended spread of gene drive mosquitoes beyond trial sites, or resistance mechanisms and non-functional effector genes that spread within trial and intervention sites. This is of particular importance for non-localized gene drive projects, as the potential scale of intervention means that monitoring is expected to be more costly than research, development and deployment. Regarding monitoring needs for population replacement systems, lessons may be learned from experiences with Wolbachia-infected mosquitoes, and for population suppression systems, from experiences with releases of genetically sterile male mosquitoes. For population suppression systems, assessing monitoring requirements for tracking population size and detecting rare resistant alleles are priorities, while for population replacement systems, allele frequencies must be tracked, and pressing concerns include detection of gene drive alleles with non-functional effector genes, and resistance of pathogens to functional effector genes. For spread to unintended areas, open questions relate to the optimal density and placement of traps and frequency of sampling in order to detect gene drive alleles, drive-resistant alleles or non-functional effector genes while they can still be effectively managed. Invasive species management programs face similar questions, and lessons may be learned from these experiences. We explore these monitoring needs for gene drive mosquito projects progressing through the phases of pre-release, release and post-release.


Induction of antiestrogen resistance in human breast cancer cells by random insertional mutagenesis using defective retroviruses: identification of bcar-1, a common integration site.
Duration of response to antiestrogen therapy in metastatic breast cancer is limited due to the development of antiestrogen-resistant tumors. The mechanisms involved are not understood but could originate from (epi)genetic alterations within the tumor cells. We have applied in vitro random insertional mutagenesis with replication defective retroviruses to identify those genes playing a key role in development of antiestrogen resistance in human breast cancer cells. Eighty antiestrogen-resistant cell clones were isolated from 7 x 10(8) estrogen-dependent ZR-75-1 cells, mass-infected with defective retroviruses and subjected to 4-OH-tamoxifen selection. Integration site-specific DNA probes were made by inverse polymerase chain reaction techniques and used to search for common integration sites. Six cell clones were identified with retroviral genome integrations in the same orientation in a single locus, designated breast cancer antiestrogen resistance locus-1 (bcar-1). These bcar-1 cell clones had lost estrogen receptor expression and had become estrogen independent. Our results strongly suggest that alteration of the bcar-1 locus is responsible for development of antiestrogen resistance in human breast cancer cells in vitro. In addition, we have shown that in vitro insertional mutagenesis using defective retroviruses can be applied for gene tagging in human cells.


Stress analysis of irradiated human tooth enamel using finite element methods.
The objectives of this project were to use finite element methods to determine how changes in the elastic modulus due to oral cancer therapeutic radiation alter the distribution of mechanical stresses in teeth and to determine if observed failures in irradiated teeth correlate with changes in mechanical stresses. A thin slice section finite element (FE) model was constructed from micro CT sections of a molar tooth using MIMICS and 3-Matic software. This model divides the tooth into three enamel regions, the dentin-enamel junction (DEJ) and dentin. The enamel elastic modulus was determined in each region using nano indentation for three experimental groups namely - control (non-radiated), in vitro irradiated (simulated radiotherapy following tooth extraction) and in vivo irradiated (extracted subsequent to oral cancer patient radiotherapy) teeth. Physiological loads were applied to the tooth models at the buccal and lingual cusp regions for all three groups (control, in vitro and in vivo). The principal tensile stress and the maximum shear stress were used to compare the results from different groups since it has been observed in previous studies that delamination of enamel from the underlying dentin was one of the major reasons for the failure of teeth following therapeutic radiation. From the FE data, we observed an increase in the principal tensile stress within the inner enamel region of in vivo irradiated teeth (9.97 ± 1.32 MPa) as compared to control/non-irradiated teeth (8.44 ± 1.57 MPa). Our model predicts that failure occurs at the inner enamel/DEJ interface due to extremely high tensile and maximum shear stresses in in vivo irradiated teeth which could be a cause of enamel delamination due to radiotherapy.


[Studies on the usefulness of saliva for detection of antibodies to HIV-1].
It has been reported that antibodies to HIV-1 could be detected in saliva of patients with AIDS. We studied whether saliva is potentially useful for screening of HIV-1 infected persons. Pairs of serum and saliva were collected from both 19 HIV-1 seropositive outpatients (CDC type, II:15, III:1, IV:3) of AIDS clinic in our hospital and 4 controls. Multiple saliva collection was done from seropositives periodically for 8 months after the first sampling. Serum and saliva were tested with ELISA and Western blot (WB) methods by using kits of diagnostics Pasteur (ELAVIA MIXT and LAV Blot-1). All pairs of serum and saliva from controls were clearly negative by ELISA. Nineteen sera of proven seropositive cases and paired 18 saliva samples were confirmed to be positive in ELISA test employed, but O.D. value of 1 saliva sample was below the cut-off level. However, in the follow-up study, samples taken from the same individuals after the first sampling showed positive results. Forty-seven saliva samples from seropositives were served for the WB test. Clearly positive bands were observed in 43 samples. In each of the remaining 5 samples, the final decision was "indeterminate", although a strong reactive band was observed at GP-160. The results mentioned above suggested that saliva was useful for screening of antibodies to HIV-1 in epidemiological studies, though it is necessary to improve the sensitivity of ELISA and WB for tests of saliva.


Pathologic findings and liver elements in hibernating bats with white-nose syndrome.
Two groups of vespertilionid bats were collected from affected hibernacula. In group 1 (n, 14; pathology and microbiology), the average body weights of all species were at the lower limit of published ranges. Twelve bats (86%) had mycotic growth in the epidermis, hair follicles, and sebaceous glands. Geomyces destructans, with its characteristic curved conidia, was observed microscopically, cultured, and confirmed by polymerase chain reaction. Dermatitis and mural folliculitis was nil to mild. When focally coinfected with Gram-negative bacteria, there was necrosis and pustules. Fat stores were little to abundant in 12 bats (86%) and nil in 2. Thirteen bats (93%) had pulmonary congestion and 7 (50%) had bone marrow granulocytosis. In group 2 (n, 24; liver elements), 3 bats (13%) had potentially toxic lead levels and 1 (4%), potentially toxic arsenic level. There was no evidence of major organ failure or consistent element toxicity.


Increased Prevalence of low High-density Lipoprotein Cholesterol (HDL-C) Levels in Korean Adults: Analysis of the Three Korean National Health and Nutrition Examination Surveys (KNHANES 1998-2005).
High-density lipoprotein cholesterol (HDL-C) is an independent risk factor for cardiovascular diseases that has shown a remarkable increase, but little is known about the prevalence of low HDL-C in Korea. This study aimed to evaluate changing trends of low HDL-C prevalence, and indicate other risk factors associated with low HDL-C. We selected subjects aged ≥20 years from the Korean National Health and Nutrition Examination Survey (KNHANES) 1998, 2001, and 2005 (n = 7962, 6436, and 6412). The mean level of HDL-C and the prevalence of low HDL-C was calculated, and cardiovascular risk factors associated with low HDL-C, as well as demographic, anthropometric, lifestyle, and nutrition factors, were assessed using the KNHANES 2005 data. Mean HDL-C levels in men and women between KNHANES 1998 and 2005 decreased significantly, from 48.1 to 42.3 and from 51.6 to 47.1 mg/dL, respectively (both p < 0.001). The decrease was slightly less for women compared with men for the same period, and women had higher HDL-C levels at all periods. Covariate-adjusted OR revealed that body mass index, waist circumference, and non-alcohol drinker in both men and women were associated with low HDL-C levels by KNHANES 2005, as were employed and light physical activity in men and low fat intake in women. The prevalence of low HDL-C increased significantly from KNHANES 1998 to 2001 and 2005 (p < 0.001) in both men and women. body mass index, waist circumference, and non-alcohol drinker were identified as associated with low HDL-C in Korean adults.


Na(+)/H(+) exchanger regulatory factor 1 (NHERF1) is required for the estradiol-dependent increase of phosphatase and tensin homolog (PTEN) protein expression.
Expression of phosphatase and tensin homologue deleted on chromosome 10 (PTEN) can be induced by estrogens at the posttranscriptional level. However, the molecular mechanism of the process is unclear. In this study, we found that the C terminus (CT) of PTEN is indispensable for 17-β-estradiol (E2)-increased PTEN expression. Therefore, we screened for PTEN-CT-associated proteins using a glutathione-S-transferase pull-down approach in combination with mass spectrometry-based proteomic analyses. Our experiments led to the identification of Na(+)/H(+) exchanger regulatory factor 1 (NHERF1) as a major PTEN-CT binding partner. The first postsynaptic density protein-95/Discslarge/zonula occludens-1 homology domain of NHERF1 and the last four amino acids of PTEN were found to be key determinants of this interaction. By associating with PTEN, NHERF1 could enhance PTEN protein expression by retention of PTEN turnover, as demonstrated by NHERF1 overexpression and small interfering RNA-mediated knockdown experiments, respectively. Furthermore, NHERF1 inhibited ubiquitination of the PTEN protein upon competition with binding of PTEN to neural precursor cell expressed, developmentally down-regulated 4, an ubiquitin E3 ligase. E2 strongly induced the expression of NHERF1 and PTEN only in estrogen receptor (ER)-positive cells but not in ER-negative cells. ICI182780, an ER-specific inhibitor, decreased the expression of both NHERF1 and PTEN, and ICI182780 pretreatment also retarded E2-increased PTEN expression in ER-MDA-MB-231 cells. In both ER-MDA-MB-231 and MCF-7 cells, E2 failed to increase PTEN expression when NHERF1 was knocked down. Taken together, these are the first results that present a possible mechanism for E2-increased PTEN expression. In this process, E2 first induces NHERF1 expression by activating the ER. Upon competition with neural precursor cell expressed, developmentally down-regulated 4, NHERF1 then interacts with PTEN to inhibit PTEN degradation, through an ubiquitination-dependent pathway. This in turn leads to the increase of PTEN expression at the protein level.


Formulating the Structural Aspects of Various Benzimidazole Cognates.
Benzimidazole derivatives are widely used in clinical practice as potential beneficial specialists. Recently, the neuroprotective effect of derivatives of benzimidazole moiety has also shown positive outcomes. To develop favourable molecules for various neurodegenerative disorders using the versatile chemical behaviour of the benzimidazole scaffold. About 25 articles were collected that discussed various benzimidazole derivatives and categorized them under various subheadings based on the targets such as BACE 1, JNK, MAO, choline esterase enzyme, oxidative stress, mitochondrial dysfunction in which they act. The structural aspects of various benzimidazole derivatives were also studied. To manage various neurodegenerative disorders, a multitargeted approach will be the most hopeful stratagem. Some benzimidazole derivatives can be considered for future studies, which are mentioned in the discussed articles.


Tripeptide IRW Upregulates NAMPT Protein Levels in Cells and Obese C57BL/6J Mice.
Nicotinamide adenine dinucleotide (NAD+) plays a vital role in cellular processes that govern human health and disease. Nicotinamide phosphoribosyltransferase (NAMPT) is a rate-limiting enzyme in NAD+ biosynthesis. Thus, boosting NAD+ level via an increase in NAMPT levels is an attractive approach for countering the effects of aging and metabolic disease. This study aimed to establish IRW (Ile-Arg-Trp), a small tripeptide derived from ovotransferrin, as a booster of NAMPT levels. Treatment of muscle (L6) cells with IRW increased intracellular NAMPT protein levels (2.2-fold, p < 0.05) and boosted NAD+ (p < 0.01). Both immunoprecipitation and recombinant NAMPT assays indicated the possible NAMPT-activating ability of IRW (p < 0.01). Similarly, IRW increased NAMPT mRNA and protein levels in the liver (2.6-fold, p < 0.01) and muscle tissues (2.3-fold, p < 0.05) of C57BL/6J mice fed with a high-fat diet (HFD). A significantly increased level of circulating NAD+ was also observed following IRW treatment (4.7 fold, p < 0.0001). Dosing of Drosophila melanogaster with IRW elevated both D-NAAM (fly NAMPT) and NAD+ in vivo (p < 0.05). However, IRW treatment did not boost NAMPT levels in SIRT1 KO cells, indicating a possible SIRT1 dependency for the pharmacological effect. Overall, these data indicate that IRW is a novel small peptide booster of the NAMPT pool.


Cytology of metastatic leiomyosarcoma. A case report.
Metastasis of leiomyosarcoma is not uncommon. Determination of the primary site may be confused with the presenting symptoms and even the known pathology. A 62-year-old man presented with chest pain, a productive cough and a lung mass on radiography. Fine needle aspiration of the tumor showed pleomorphic tumor cells. The patient began to have abdominal symptoms one year after presentation. Jejunal leiomyosarcoma was proven after an emergency exploratory laparotomy for mechanical ileus. Sputum cytology disclosed tumor cells one month before death. Necropsy of the lung mass demonstrated findings similar to those of the intestinal tumor microscopically. Without laparotomy, our patient would have been diagnosed as having a primary pulmonary leiomyosarcoma by the presenting symptoms and known pathology.


Localized laryngeal amyloidosis - a case report.
Amyloidosis encompasses a variety of conditions, caused by extracellular, insoluble protein fibrils that disturb the normal functioning of cells and organs. The disease may be localized or systemic, hereditary or acquired (associated with chronic inflammatory or hematological diseases). We present the case of a 49-year-old woman, with symptoms including dysphagia, dysphonia and dyspnea. After taking the case history and performing clinical examination, we suspected a laryngeal tumor to be the cause of the symptoms. Microlaryngoscopy and biopsy were performed. The histopathological examination result of the biopsy specimen was amyloidosis. Surgical excision of the tumor was performed. Our case presentation describes this rare pathological finding, its clinical manifestations, the histopathological and surgical diagnostic problems, treatment, patient evolution and the difficulties we encountered along the way, through the scope of our personal experience.


[Radiation therapy in locally advanced and/or relapsed urological tumors].
Radiation therapy (RT) plays a fundamental role in the treatment of locally advanced and/or relapsed urological tumors, as well as in palliation, or as definitive treatment, and even where integrated into a multi-modal approach. In operated renal tumors, positive margins or extracapsular extension show a positive impact of postoperative RT, with a reduction of relapses between 100% and 30%, while, in the case of palliation, treatments with RT at high doses are preferred. In advanced cancers of the upper urinary tract, RT plays a limited role, even if it seems to increase the level of disease control locally and, with the combination of cisplatin, survival rates too. An important reduction in the recurrence is also observed in locally advanced tumors of the urethra, with a recurrence of 60% after surgery, 36% after RT and 25% after pairing of the two. In locally advanced tumors of the penis, RT shows poorer results than surgery, and the addition of postoperative RT does not seem to add any further outcome, except where, in the presence of a positive inguinal dissection, the postoperative RT reduces lymph node recurrences by 60%-11%. Interesting data for the preservation of the organ are reported with reference to the combination with chemotherapy. In the tumors of the testis, it is still disputable whether the treatment of residual masses after chemotherapy may be appropriate, with a view to a possible salvage radiotherapy. In the treatment of the prostate, the role of RT is consolidated and evolving with the progress of dose escalation, the association with hormonal therapy, new technologies, new possibilities of IMRT and proton therapy and various studies on multi-modal approaches (hormone therapy, surgery, radiotherapy, chemotherapy). Cystectomy is the gold standard for the treatment of locally advanced bladder cancer, even though there is a revived interest in multimodal treatments (transurethral resection, chemotherapy, RT) that may allow the organ preservation. Postoperative radiotherapy, which can reduce by 50% to 20%-5% local recurrences that are highly correlated with distance failure and with survival, should be revised in the light of modern RT techniques that can further increase local control levels and reduce the toxicity significantly.


[Experience of radiofrequency ablation therapy for hepatocellular carcinoma].
There are various therapeutic options for hepatocellular carcinoma. Radiofrequency ablation (RFA) was introduced to Japan in 1998, and has become popular in percutaneous local treatment for HCC as a treatment with the advantage of both percutaneous ethanol injection therapy and percutaneous microwave coagulation therapy. In this study, we investigated the efficacy and complication of RFA for HCC. Seventeen patients underwent percutaneous or open RFA with a Cool-tip needle (Radionics Co. Ltd) from April 2001 to May 2005. All tumors were solitary and the average diameter of 17 tumors was 2.24 cm. Fifteen tumors were completely ablated, but two weren't. Local recurrence occurred in only one patient and the rate of local recurrence was 6.7%. Mild complication occurred in some patients, but critical complication did not occur in any patients. In this study, it was considered that RFA could be performed safely and was a good treatment for HCC with high efficacy. After investigating the long-term results and indication of RFA, it was suggested that RFA might be reestablished as an effective treatment for HCC.


Stakeholder issues for the physician executive.
If physician executives are to cope with the environmental turbulence and uncertainty facing their organizations, they must effectively manage their "stakeholders." The stakeholder approach helps integrate managerial concerns that are frequently treated separately, such as strategic management, marketing, human resource management, "organizational politics," and social responsibility. The stakeholder perspective enables medical managers to relate important issues to the development of strategies for handling potentially conflicting demands for effectiveness and efficiency from various stakeholders. Medical managers should minimally satisfy the needs of marginal stakeholders while they maximally satisfy the needs of key stakeholders. To identify key stakeholders, a physician executive should critically assess each stakeholder's potential to threaten the organization and its potential to cooperate. This assessment should account for such factors as the stakeholder's relative power, the specific context and history of the organization's relations with it, the specific issues under consideration, and other key stakeholders influencing the organization.


Twisting of fibrin fibers limits their radial growth.
Electron microscopy of freeze-dried, shadowed fibrin fibers has demonstrated that these structures are twisted. The pitch and radius of many fibers were measured from the micrographs. Although there is some variability, the average pitch of 1930 +/- 280 (SD) nm is independent of radius. The distribution of observed radii of fibers assembled in vitro is highly skewed, suggesting that individual fibers grow to a maximum radius of about 50 nm, except when both pH and ionic strength are high; fibers aggregate to form thicker fiber bundles under some conditions. The observed twisting may be responsible for limiting the lateral growth of individual fibers. Protofibrils near the surface of a twisted fiber are stretched relative to those near the center. Consequently, the degree to which a protofibril can be stretched limits the radius of a fiber; protofibrils can be added to a growing fiber until the energy required to stretch an added protofibril exceeds the energy of binding. These properties of assembly arise directly from the intrinsic twist of the fibrinogen molecule determined from structural evidence. Simple geometric considerations lead to conclusions regarding the locations of the binding sites for assembly of the protofibril and the flexibility of the fibrin molecule.


Splitting of the umbilical cord in a 13-week-old fetus.
There is an increasing interest in the physiology and pathology of the umbilical cord because it is recognized as an important source of placental and, consequently, fetal problems. During the postmortem examination of a severely macerated 13-week-old fetus, a split umbilical cord was noted. This rare finding was seen in the middle segment of the cord, the fetal and placental ends both being normal. The pathogenesis of this lesion is not fully understood, and it is possible that it results through focal degeneration of previously formed Wharton's jelly or secondary loss of Wharton's jelly due to incomplete fusion or hypoplasia of the amniotic covering. Whatever the pathogenesis, it is assumed that an umbilical vessel devoid of its protective Wharton's jelly is more prone to compression and thrombosis with all its deleterious effects. Death in this case was probably associated with the congenital heart defect also presented by the fetus. The rarity of this lesion is probably explained by the fact that it represents the end of the spectrum of longitudinal deficiency of Wharton's jelly, a relatively common finding.


Mitochondrial Transplantation Attenuates Cerebral Ischemia-Reperfusion Injury: Possible Involvement of Mitochondrial Component Separation.
Mitochondrial dysfunctions play a pivotal role in cerebral ischemia-reperfusion (I/R) injury. Although mitochondrial transplantation has been recently explored for the treatment of cerebral I/R injury, the underlying mechanisms and fate of transplanted mitochondria are still poorly understood. Mitochondrial morphology and function were assessed by fluorescent staining, electron microscopy, JC-1, PCR, mitochondrial stress testing, and metabolomics. Therapeutic effects of mitochondria were evaluated by cell viability, reactive oxygen species (ROS), and apoptosis levels in a cellular hypoxia-reoxygenation model. Rat middle cerebral artery occlusion model was applied to assess the mitochondrial therapy in vivo. Transcriptomics was performed to explore the underlying mechanisms. Mitochondrial fate tracking was implemented by a variety of fluorescent labeling methods. Neuro-2a (N2a) cell-derived mitochondria had higher mitochondrial membrane potential, more active oxidative respiration capacity, and less mitochondrial DNA copy number. Exogenous mitochondrial transplantation increased cellular viability in an oxygen-dependent manner, decreased ROS and apoptosis levels, improved neurobehavioral deficits, and reduced infarct size. Transcriptomic data showed that the differential gene enrichment pathways are associated with metabolism, especially lipid metabolism. Mitochondrial tracking indicated specific parts of the exogenous mitochondria fused with the mitochondria of the host cell, and others were incorporated into lysosomes. This process occurred at the beginning of internalization and its efficiency is related to intercellular connection. Mitochondrial transplantation may attenuate cerebral I/R injury. The mechanism may be related to mitochondrial component separation, altering cellular metabolism, reducing ROS, and apoptosis in an oxygen-dependent manner. The way of isolated mitochondrial transfer into the cell may be related to intercellular connection.


Enhanced thrombin and plasmin activity with exercise in man.
We measured products of thrombin and plasmin action and of the platelet release reaction during exercise to determine if the well-known effect of exercise on in vitro coagulation and fibrinolytic tests reflects activity of these systems in vivo. Plasma fibrinopeptide A, produced by thrombin-mediated proteolysis of fibrinogen, increased with graded treadmill and cycle exercise to postexercise levels of 20--30 times resting values. Fibrin/fibrinogen-related D antigen increased in a similar fashion with peak levels at maximal O2 uptake. Plasma-activated partial thromboplastin times fell as fibrinopeptide A levels increased. Unheated fibrin plate lysis areas increased as D antigen concentrations rose, indicating increased release of plasminogen activator. In contrast to activation of the soluble coagulation and fibrinolytic systems, platelet counts and plasma levels of beta-thromboglobulin, a platelet release protein, did not change significantly with exercise. The effect of exercise on thrombin and plasmin was not influenced by prior physical training, but appeared to be less with cycle exercise than with treadmill exercise.


Exploring the Pharmaceutical Care of Pharmacists in China During COVID-19-A National Multicenter Qualitative Study.
Pharmacists are one of the coronavirus disease-2019 (COVID-19) treatment team members in China, yet only a few single-center studies have described the work experience of pharmacists during the pandemic. This study aimed to explore in-depth experiences of hospital pharmacists providing pharmacy services during the COVID-19 pandemic in China on a national scale. This is a nationwide multicenter qualitative study that used the purposive sampling method. Semi-structured interviews were conducted with 11 pharmacists from large-scale tertiary hospitals in representative provinces of mainland China. The Colaizzi seven-step method was applied to analyze the interview data. Eleven semi-structured interviews were conducted. Each interview lasted 25-70 min. By analyzing the work experiences of pharmacists in COVID-19 designated treatment hospitals, five descriptive themes were categorized: (1) drug supply service; (2) routine clinical pharmacy services; (3) expanded pharmacy services during the epidemic; (4) drug management loopholes; (5) areas of improvements of pharmacy services during a pandemic. During the COVID-19 epidemic, Chinese hospital pharmacists played various vital roles. However, there were loopholes in managing narcotic drugs, psychotropic drugs, and donated drugs. The study uncovered areas of improvement in pharmacy services during the pandemic. The emergency response capacity of hospital pharmacists should be continuously improved in the future.


Applicability of assessment center concepts and techniques for managerial selection and development in human service organizations.
The Assessment Center technique has been used widely throughout private industry over the past twenty-five years for selection and promotion of supervisory and managerial personnel. It has been acclaimed as a fair and valid method and has been successful in identifying individuals who possess the greatest potential to achieve in a particular position. A growing interest in the field of human services administration is the need to find the best person to select or promote into a supervisory or administrative position. The Assessment Center can be used to address this need, and the authors discuss the applicability of the technique to the human service organization.


[Large cell anaplastic lymphomas in the immunocompromised host].
Large cell anaplastic (LCA) lymphomas are a newly defined tumor entity, which has recently been integrated in the updated Kiel classification. The occurrence of CD30+ LCA lymphoma in the setting of renal transplant patients has so far been reported only once. This report describes two LCA lymphomas of B-cell phenotype in renal transplant patients. The clonal evolution and possible etiologic role of Epstein-Barr Virus (EBV) in LCA lymphoma was studied. Our findings give further evidence that clonal evolution of B-cell populations is a second event in lymphomagenesis and is, at least in the cases studied, preceeded by either reactivated latent or primary EBV-infection with clonal EBV proliferation.


Echocardiography without electrocardiogram.
automatic detection of the QRS complex on electrocardiogram (ECG) is used on cardiac ultrasound scanners to separate ultrasound image series into cardiac cycles for playback and storage. On small hand-held scanners it is unpractical to connect ECG cables. We therefore aim to do automatic cardiac cycle separation using apical B-mode ultrasound images. cardiac cycle length is estimated by cyclicity analysis of B-mode intensities. To determine a cycle start estimate near QRS, a deformable model is fitted to the left ventricle in real-time. The model is used to initialize and constrain a speckle tracker positioned near the mitral annulus. In the displacement curve generated by the speckle tracker, a time point near maximum distance from the probe is detected as a cardiac cycle start estimate. Validation against ECG was done on 233 recordings from normal subjects and 46 recordings from subjects with coronary pathology. Several test cases were run for each recording to emulate B-mode series starting at all time points in the cardiac cycle. Totally, 11 886 test cases were run. Cycle length estimation was feasible in 98% of normal subject cases and 91% of pathology cases. Median difference in cycle length by ECG was 0 and -3 ms, respectively. Cycle start estimation was feasible in 90% of normal subject cases and 77% of pathology cases. Median difference to cycle start by ECG was 62 and 76 ms, respectively. apical B-mode series can automatically be separated into cardiac cycles without using ECG.


The retention behaviour of amino acids in hydrophilic interaction liquid chromatography on zwitterionic stationary phases.
The retention behaviour of amino acids was studied in hydrophilic LC on zwitterionic stationary phases. Evaluation of the influences of acetonitrile/water content, ammonium acetate (NH4Ac) concentration and mobile phase pH values was performed. Fourteen amino acids were tested and they were all retained to varying extents, with poorer retention in high water content eluents. The linear relationship between the logarithm of retention factor and log(water content) indicated that adsorption dominated or at least was partly involved in the separation mechanism. Electrostatic and hydrophilic interactions also contributed to the retention of these amino acids under different separation conditions with various mobile phase pH values and NH4Ac concentrations. Thus, the overall retention mechanism could be explained as a combination of adsorption, electrostatic and hydrophilic interactions. The magnitude and contribution of each mechanism is dependent on the nature of the analyte and the separation conditions applied.


Impact of depression and post-traumatic stress disorder on functional outcome and health-related quality of life of patients with mild traumatic brain injury.
The impact of disability following traumatic brain injury (TBI), assessed by functional measurement scales for TBI or by health-related quality of life (HRQoL), may vary because of a number of factors, including presence of depression or post-traumatic stress disorder (PTSD). The aim of this study was to assess prevalence and impact of depression and PTSD on functional outcome and HRQoL six and 12 months following mild TBI. We selected a sample of 1919 TBI patients who presented to the emergency department (ED) followed by either hospital admission or discharge to the home environment. The sample received postal questionnaires six and 12 months after treatment at the ED. The questionnaires included items regarding socio-demographics, the 36-item Short-Form Health Survey (SF-36), the Perceived Quality of Life Scale (PQoL), the Beck Depression Inventory, and the Impact of Event Scale. A total of 797 (42%) TBI patients completed the six-month follow-up survey. Depression and PTSD prevalence rates at both the six- and 12-month follow-up were 7% and 9%, respectively. Living alone was an independent predictor of depression and/or PTSD at six- and 12-month follow-up. Depression and PTSD were associated with a significantly decreased functional outcome (measured with Glasgow Outcome Scale Extended) and HRQoL (measured using the SF-36 and the PQoL). We conclude that depression and/or PTSD are relatively common in our sample of TBI patients and associated with a considerable decrease in functional outcome and HRQoL.


Circular on illiteracy, 1988.
In 1988, the Chinese State Council issued a Circular calling for a nationwide campaign to do away with illiteracy within five years. The goal of the campaign is to ensure that over 85% of the population in rural areas and over 90% of the population in enterprises, institutions, and cities between the ages of 15 and 40 is literate. The campaign calls for governments at all levels to take measures against illiteracy and requires peasants to learn at least 1500 characters and industrial workers and urban residents to learn 2000 characters. Certificates are to be issued to persons who meet the standards.


[Research on particle size and size distribution of nanocrystals in urines by laser light scattering method].
In the present paper laser light scattering method was used to investigate the particle size and size distribution of nanoparticles simultaneously in urines of lithogenic patients and healthy persons. This method is economic, rapid, accurate and easy to operate. The results showed that healthy urines are more stable than lithogenic urines. In urines of healthy human, the ultrafine crystals were well scattered and not aggregated with a smaller size. However, the ultrafine crystals in lithogenic urine have a broad size distribution, which increases the aggregation trend of nanocrystals. Based on the intensity-autocorrelation curve, the stability of urine samples of both healthy human and lithogenic patients was comparatively investigated. The relationship between the measurement results and the methods of handling sample was studied. The results show that a stable urine sample can be obtained by diluting the urine with a ratio of 20%, then centrifuging it at 4,000 round per minute for 15 minutes or filtrating it with 1.2 microm cellulose acetate filter. The results of laser light scattering method are consistent with that obtained by transmission electron microscopy (TEM). The reasons for the stability of urines are explained from the points of Van der Waals force, urine viscosity, pH value, ionic strength, surface charge and zeta potential of the ultrafine crystals, and so on. The results in this paper provide a new thought for preventing formation and recurrence of urinary stones.


Use of population-weighted Estimated Average Requirements as a basis for Daily Values on food labels.
In both Canada and the United States, nutrition labeling is now mandatory for most packaged foods. The labeling is intended to help consumers select foods that can contribute to a healthful diet, but current label values are based on outdated notions of nutrient requirements. The Committee on Use of Dietary Reference Intakes in Nutrition Labeling has recommended that the reference values used for nutrition labeling be based on a population-weighted Estimated Average Requirement (EAR) for any nutrient for which requirements have been estimated. This value approximates the median of the distribution of nutrient requirements for individuals who are members of the target population for food labels. It provides the most scientifically valid, single point of comparison for an appraisal of the probable contribution of a specific food to the overall nutrient needs of individuals in the target population. In contrast, a reference value based on a population coverage approach would understate the nutrient contribution of the food item relative to the requirements of the vast majority of individuals in the target population and thus offer misinformation rather than positive guidance to the consumer.


Morphine desensitization, internalization, and down-regulation of the mu opioid receptor is facilitated by serotonin 5-hydroxytryptamine2A receptor coactivation.
Analysis of the distribution of mRNA encoding the serotonin (5-hydroxytryptamine) 5-HT(2A) receptor and the mu opioid peptide receptor in rat brain demonstrated their coexpression in neurons in several distinct regions. These regions included the periaqueductal gray, an area that plays an important role in morphine-induced analgesia but also in the development of tolerance to morphine. To explore potential cross-regulation between these G protein-coupled receptors, the human mu opioid peptide receptor was expressed stably and constitutively in Flp-In T-REx human embryonic kidney 293 cells that harbored the human 5-HT(2A) receptor at the inducible Flp-In locus. In the absence of the 5-HT(2A) receptor, pretreatment with the enkephalin agonist [D-Ala(2),N-Me-Phe(4),Gly(5)-ol]-enkephalin but not with the alkaloid agonist morphine produced desensitization, internalization, and down-regulation of the mu opioid peptide receptor. Induction of 5-HT(2A) receptor expression in these cells resulted in up-regulation of mu opioid peptide receptor levels that was blocked by both a 5-HT(2A) receptor inverse agonist and selective inhibition of signaling via Galpha(q)/Galpha(11) G proteins. After induction of the 5-HT(2A) receptor, coaddition of 5-HT with morphine now also resulted in desensitization, receptor internalization, and down-regulation of the mu opioid peptide receptor. It has been argued that enhancement of mu opioid peptide receptor internalization in response to morphine would limit the development of tolerance without limiting analgesia. These data suggest that selective activation of the 5-HT(2A) receptor in concert with treatment with morphine might achieve this aim.


Influence of growth phase on the essential oil composition of Ziziphora clinopodioides Lam.
Ziziphora clinopodioides belongs to the Lamiaceae family, that are widespread all over Iran. The leaves, flowers and stems of the plant are frequently used as wild vegetables or additives in food to offer aroma and flavour. The chemical composition of the essential oils obtained from the aerial parts of Z. clinopodioides Lam. in different growth stages were analysed by GC and GC-MS, and 29 components were identified. Pulegone (30.1%), thymol (21.3%), p-mentha-3-en-8-ol (12.9%) and piperitenone (9.3%) were the main components in the pre-flowering stage. In the flowering stage, pulegone (44.6%), p-mentha-3-en-8-ol (10.5%), 1,8-cineoil (10.4%), piperitenone (8.7%) and thymol (6.7%) were identified as the main constituents and in the post-flowering stage, pulegone (41.3%), isomenthone (11.6%), p-mentha-3-en-8-ol (11%), p-mentha-3,8-diene (7.2%) and thymol (5.8%) were identified as the major components.


Strategies Using Gelatin Microparticles for Regenerative Therapy and Drug Screening Applications.
Gelatin, a denatured form of collagen, is an attractive biomaterial for biotechnology. In particular, gelatin particles have been noted due to their attractive properties as drug carriers. The drug release from gelatin particles can be easily controlled by the crosslinking degree of gelatin molecule, responding to the purpose of the research. The gelatin particles capable of drug release are effective in wound healing, drug screening models. For example, a sustained release of growth factors for tissue regeneration at the injured sites can heal a wound. In the case of the drug screening model, a tissue-like model composed of cells with high activity by the sustained release of drug or growth factor provides reliable results of drug effects. Gelatin particles are effective in drug delivery and the culture of spheroids or cell sheets because the particles prevent hypoxia-derived cell death. This review introduces recent research on gelatin microparticles-based strategies for regenerative therapy and drug screening models.


Development of a cell-line model to mimic the pro-survival effect of nurse-like cells in chronic lymphocytic leukemia.
The interaction between Chronic lymphocytic leukemia (CLL) cells and monocyte-derived nurse-like cells (NLCs) is fundamentally important to CLL biology. However, studies of how CLL cells and NLCs interact have been hampered by the need for freshly obtained CLL blood samples, coupled with wide variation in the number of monocytes present in the blood of individual patients. Here, we report the development and validation of a cell-line model of NLCs which overcomes these difficulties. Co-culture of primary CLL cells with THP-1 cells induced to differentiate into macrophages by phorbol 12-myristate 13-acetate (PMA) significantly reduced both spontaneous and fludarabine-induced cell death of leukemic cells. Furthermore, compared with their M1-polarized counterparts, M2-polarized macrophages derived from PMA-differentiated THP-1 cells conferred to CLL cells greater protection from spontaneous and fludarabine-induced apoptosis. Since NLCs resemble M2 tumor-associated macrophages, this cell-line model could be useful for investigating the mechanisms through which NLCs protect CLL cells from spontaneous and drug-induced apoptosis.


Structure of solid tumors and their vasculature: implications for therapy with monoclonal antibodies.
Delivery of monoclonal antibodies to solid tumors is a vexing problem that must be solved if these antibodies are to realize their promise in therapy. Such success as has been achieved with monoclonal antibodies is attributable to the local hyperpermeability of the tumor vasculature, a property that favors antibody extravasation at tumor sites and that is mediated by a tumor-secreted vascular permeability factor. However, leaky tumor blood vessels are generally some distance removed from target tumor cells, separated by stroma and by other tumor cells that together represent significant barriers to penetration by extravasated monoclonal antibodies. For this reason, alternative approaches may be attractive. These include the use of antibody-linked cytotoxins, which are able to kill tumor cells without immediate contact, and direction of antibodies against nontumor cell targets, for example, antigens unique to the tumor vascular endothelium or to tumor stroma.


Trypanosoma cruzi Infection Impairs the Endocytosis of Zymosan A by Cardiomyocytes.
We have previously reported that mannose receptors participate and are regulated during Trypanosoma cruzi cardiomyocyte (CM) infection. Our present aim is to characterize the endocytosis of mannosylated ligands like zymosan A (Zy) in uninfected and T. cruzi-infected CM. CM infected or not by T. cruzi were incubated with Zy for different periods of time and their internalization was analyzed at light microscopy level. Fluorescent approaches were performed by treating Zy with concanavalin-A-TRITC and washing it exhaustively prior to incubation with CM. The cultures were further stained with phalloidin-FITC and DAPI for actin and DNA visualization, respectively. CM internalized Zy particles in a time-dependent fashion. The ligand specificity was confirmed by the addition of mannan, which efficiently blocked the Zy endocytosis. Designed fluorescent approaches extended and confirmed the Zy internalization by striated cells. Infected cultures displayed impairment in Zy endocytosis, which seems to be directly related to host infection rates. Altogether, our results show the ability of CM to ingest large particles such as the mannosylated ligand Zy. During their infection with T. cruzi, there is a loss in Zy internalization possibly due to the negative modulation of mannose receptors.


Nonlinear optoacoustic transformation in the system of dielectric substrate/submicron metal coating/liquid.
The optoacoustic method has been shown to be an accurate technique for the measurement of the properties of submicron metal coatings deposited on a dielectric substrate, i.e., mirrors. The method has been previously theoretically described in terms of a linear model of optoacoustic transformation in a system substrate/coating/liquid. The goal of the present work was to determine the limits at which the linear model is still applicable. The modification of the laser induced acoustic signal profiles and transfer functions of optoacoustic transformation versus the laser fluence was studied for two liquids: ethanol and water.


Anatomic guidance for ablation: atrial flutter, fibrillation, and outflow tract ventricular tachycardia.
After initial documentation of excellent efficacy with radiofrequency ablation, this procedure is being performed increasingly in more complex situations and for more difficult arrhythmia. In these circumstances, an accurate knowledge of the anatomic basis for the ablation procedure will help maintain this efficacy and improve safety. In this review, we discuss the relevant anatomy for electrophysiology interventions for typical right atrial flutter, atrial fibrillation, and outflow tract ventricular tachycardia. In the pediatric population, maintaining safety is a greater challenge, and here again, knowing the neighboring and regional anatomy of the arrhythmogenic substrate for these arrhythmias may go a long way in preventing complications.


European Zebrafish Meeting 2015 Husbandry Session Report.
A workshop to address husbandry and animal welfare was held during the 9th European Zebrafish Meeting in Oslo, Norway, from June 28 to July 2, 2015. The husbandry workshop took place on Monday, June 29, and was well attended by ∼100 audience members. It highlighted problems arising from the diversity of current husbandry practices and included presentations on recent efforts to find common husbandry and animal welfare standards from a variety of international contributors, from Norway, Portugal, the United Kingdom, as well as the United States and Japan. Presentations included zebrafish and medaka as representatives of aquatic species used in biomedical research and addressed a diverse range of topics such as proposed European guidelines for zebrafish husbandry, general fish facility health and husbandry standards, cryopreservation, publication standards, and feeding strategies. The workshop highlighted the desire to develop common husbandry standards for the aquatic research community across the world.


[Pathological gambling: risk factors].
In France, consumption of gambling games increased by 148% between 1960 and 2005. In 2004, gamblers lost approximately 0.9% of household income, compared to 0.4% in 1960. This represents approximately 134 Euros per year and per head. In spite of this important increase, the level remains lower than the European average (1%). However, gambling practices may continue to escalate in France in the next few years, particularly with the recent announce of the legalisation of online games and sports betting. With the spread of legalised gambling, pathological gambling rates may increase in France in the next years, in response to more widely available and more attractive gambling opportunities. In this context, there is a need for better understanding of the risk factors that are implicated in the development and maintenance of pathological gambling. This paper briefly describes the major risk factors for pathological gambling by examining the recent published literature available during the first quarter of 2008. This documentary basis was collected by Inserm for the collective expert report procedure on Gambling (contexts and addictions). Seventy-two articles focusing on risk factors for pathological gambling were considered in this review. Only 47 of them were taken into account for analysis. The selection of these 47 publications was based on the guide on literature analysis established by the French National Agency for Accreditation and Assessment in Health (ANAES, 2000). Some publications from more recent literature have also been added, mostly about Internet gambling. We identify three major types of risk factors implicated in gambling problems: some of them are related to the subject (individual factors), others are related to the object of the addiction, here the gambling activity by itself (structural factors), and the last are related to environment (contextual or situational factors). Thus, the development and maintenance of pathological gambling seems to be conditioned by the interaction of a person and a gambling activity, in a particular context. This conceptual model is based on the well-known theory of Olivenstein on toxicomania, which was proposed in the seventies. The structural factors that appeared to be highly related to pathological gambling development and maintenance are payment modality, entertaining dimension, temporality, reward level, educational messages, gambling ambiance, gambling medium and part of hazard. Among contextual factors, availability and accessibility of gambling opportunities are well known. However, social and economic factors (e.g. culture, ethnicity, religion, education) are also important. Lastly, among individual factors, psychosocial factors are gender, age, familial and personal antecedents and psychiatric comorbidities. Neurobiological factors are not discussed here. This paper briefly summarises risk factors for development and maintenance of pathological gambling. It highlights that these factors are very similar to those that are implicated in substance use disorders, except for the gambling activity and context. Prevention regarding behavioural addictions should, in the future, take advantage of the findings on substance use disorder, since behavioural addictions are less known. Conversely, it seems obvious that findings on pathological gambling, and more widely behavioural addictions, could highlight all types of addiction. It appears that this fact is especially valid with regards to the evolution and stability of addictive status, which must be taken into account for treatment proposals.


Increasing the success of reverse sural flap from proximal part of posterior calf for traumatic foot and ankle reconstruction: patient selection and surgical refinement.
In this report, we present our experience on the use of the reverse sural flap for traumatic foot and ankle reconstruction. The patient selection and surgical refinement are discussed. From 2007 to 2010, 11 consecutive patients underwent modified reverse sural flap at the Chang Gung Memorial Hospital. The defects were located at the ankle (three cases), foot (two cases), and heel (six cases). Particular attention was paid to precise patient selection and surgical refinements. Patient selection was based on the lower limb vascular status by palpable distal pedal pulses and ankle brachial index ranging from 0.9 to 1.2. Surgical techniques were refined as precisely locating the perforators of peroneal artery, placing the skin paddle in upper third of leg for a distal region coverage, designing a 7-cm-wide adipofascial pedicle with a 2 cm skin paddle on it, preserving the mesentery structure of sural nerve and concomitant artery with or without including gastrocnemius muscles cuff, no tunneling when inset this flap and supercharging with lesser saphenous vein whenever needed. All the flaps survived completely. Only one patient required immediate anastomosis of lesser saphenous vein to local vein around defect in order to relieve the venous congestion during operation. Patients felt diminished but adequate recovery of sense of touch and temperature at the flap. Following the precise patient selection and surgical refinements, the modified reverse sural flap seemed to be a reliable and effective local flap for reconstruction of the soft tissue defects on ankle and foot.


Exonucleolytic degradation of high-density labeled DNA studied by fluorescence correlation spectroscopy.
The exonucleolytic degradation of high-density labeled DNA by exonuclease III was monitored using two-color fluorescence correlation spectroscopy (FCS). One strand of the double stranded template DNA was labeled on either one or two base types and additionally at one end via a 5' Cy5 tagged primer. Exonucleolytic degradation was followed via the diffusion time, the brightness of the remaining DNA as well as the concentration of released labeled bases. We found a hydrolyzation rate of about 11 to 17 nucleotides per minute per enzyme (nt/min/enzyme) for high-density labeled DNA, which is by a factor of about 4 slower than for unlabeled DNA. The exonucleolytic degradation of a 488 base pair long double stranded DNA resulted in a short double stranded DNA segment of 112 ± 40 base pairs (bp) length with two single-stranded tails.


Subcellular distribution of zinc in Daphnia magna and implication for toxicity.
We examined the subcellular partitioning of zinc (Zn) in Daphnia magna both under acute and chronic exposures. In the acute Zn toxicity tests, the daphnids were exposed to different Zn concentrations for 48 h or to one lethal concentration (1,000 microg/L) for different durations (time to death for up to 47 h). Significant mortality of daphnids was observed when the newly accumulated Zn concentration reached a threshold level of approximately 40 microg/g wet weight (or 320 microg/g dry wt), approximately 3.5 times higher than the background tissue concentration (92 microg/g dry wt). Chronic exposure (14 d) to Zn resulted in nonobservable effect on survivorship and growth at newly accumulated tissue concentration of over 40 microg/g wet weight. With increasing Zn acute exposure, more Zn was partitioned into the cellular debris fraction, indicating that this fraction was presumably the first targeted site of binding for Zn upon entering the animals. The importance of other subcellular fractions either decreased accordingly or remained comparable. We found that the metal-sensitive fraction (Zn distribution in the organelles and heat-denatured proteins) did not predict the acute Zn toxicity in Daphnia. During chronic exposure, however, no major change of the subcellular partitioning of Zn with increasing Zn exposure was documented. Zinc was mainly found in the organelles and heat-stable protein fractions during chronic exposure, suggesting that any subcellular repartitioning occurred primarily during acute exposure. Metallothioneins were induced upon chronic Zn exposure, but its induction evidently lagged behind the Zn accumulation. Our present study showed that the subcellular fractionation approach could not be readily used to predict the acute and chronic toxicities of Zn in Daphnia. A tissue-based Zn accumulation approach with a threshold Zn tissue concentration was better in predicting acute Zn toxicity.


Interaction of taurine with methionine: inhibition of myocardial phospholipid methyltransferase.
Perfusion of isolated, working rat heart with buffer containing 300 microM methionine in the presence of 2.5 U/L insulin led to a 15% decrease in cardiac work and a four-fold decrease in sarcolemmal Na(+)-Ca2+ exchange activity. These effects of methionine were largely prevented by inclusion of 10 mM taurine in the buffer supplemented with methionine and insulin. Taurine also reduced the extent of 3H-methyl group incorporation from radioactive methionine into myocardial phospholipids by approximately 45%. Assays of sarcolemmal phospholipid methyltransferase activity at catalytic sites I, II, and III revealed that taurine inhibited N-methylation activity approximately 30%. The data imply that the ability of taurine to modulate myocardial contraction and calcium transport may be related to taurine-mediated inhibition of phospholipid N-methylation.


Role of polymorphisms in the TNFRSF13B (TACI) gene in Spanish patients with immunoglobulin A deficiency.
Mutations in the TNFRSF13B (TACI) gene have been associated with common variable immunodeficiency, and a role in immunoglobulin A deficiency (IgAD) has also been suggested. We aimed at studying the role of several polymorphisms along this gene in IgAD susceptibility. Three TNFRSF13B mutations (C104R, A181E and R202H) and eight additional single nucleotide polymorphisms in the gene were genotyped in 338 Spanish IgAD patients and 553 ethnically matched healthy controls and tested for association. Data from parents of 114 IgAD patients were also collected and used for additional analysis. No statistically significant differences were observed after comparing patients and controls for any single nucleotide polymorphism analysed. Therefore, our work seems to discard a role of TNFRSF13B mutations in IgAD, concordantly with the most recent published studies.


Discriminative motif analysis of high-throughput dataset.
High-throughput ChIP-seq studies typically identify thousands of peaks for a single transcription factor (TF). It is common for traditional motif discovery tools to predict motifs that are statistically significant against a naïve background distribution but are of questionable biological relevance. We describe a simple yet effective algorithm for discovering differential motifs between two sequence datasets that is effective in eliminating systematic biases and scalable to large datasets. Tested on 207 ENCODE ChIP-seq datasets, our method identifies correct motifs in 78% of the datasets with known motifs, demonstrating improvement in both accuracy and efficiency compared with DREME, another state-of-art discriminative motif discovery tool. More interestingly, on the remaining more challenging datasets, we identify common technical or biological factors that compromise the motif search results and use advanced features of our tool to control for these factors. We also present case studies demonstrating the ability of our method to detect single base pair differences in DNA specificity of two similar TFs. Lastly, we demonstrate discovery of key TF motifs involved in tissue specification by examination of high-throughput DNase accessibility data. The motifRG package is publically available via the bioconductor repository. email@example.com Supplementary data are available at Bioinformatics online.


Effects of Tocilizumab, an Anti-Interleukin-6 Receptor Antibody, on Serum Lipid and Adipokine Levels in Patients with Rheumatoid Arthritis.
Patients with rheumatoid arthritis (RA) are at increased risk of cardiovascular disease. Dyslipidemia is a known adverse effect of tocilizumab (TCZ), an anti-interleukin-6 receptor antibody used in RA treatment. We aimed to assess the effect of TCZ on lipid profile and adipokine levels in RA patients. Height, weight, disease activity scores, lipid profile and atherogenic indices (AI), leptin, adiponectin, resistin, interleukin-6, and high-sensitivity C-reactive protein (CRP) were measured before and four months after initiation of TCZ in 40 RA patients and 40 healthy controls. Following TCZ treatment, total cholesterol, high density lipoprotein (HDL), and triglycerides were significantly elevated, but no significant changes in weight, body mass index (BMI), low density lipoprotein (LDL), and AI were observed. Compared with controls, significantly higher adiponectin levels were measured in the RA group at baseline. Following TCZ treatment, resistin levels and the leptin-to-adiponectin ratio increased, adiponectin levels decreased, and leptin levels remained unchanged. No correlation was found between the change in adipokine serum levels and changes in the disease activity indices, nor the lipid profile. In conclusion, the changes observed suggest a protective role for TCZ on the metabolic and cardiovascular burden associated with RA, but does not provide a mechanistic explanation for this phenomenon.


Artificial nutritional support in clinical practice in Britain.
Malnutrition is common in hospitalised patients. It often develops insidiously and its diagnosis is frequently delayed or missed. A multidisciplinary nutrition support team can improve the quality of nutritional support, reduce inappropriate feeding, reduce the complications associated with enteral and parenteral nutrition, and so improve clinical outcome and reduce hospitalisation. These improvements have obvious financial advantages, yet only a minority of British hospitals (25-30%) has a nutrition team. The increasing use of parenteral and enteral nutrition at home, which represents one of the most important areas of recent developments in artificial nutritional support, also has financial and clinical advantages, but the management of such patients is also less than optimal. Better education and greater awareness of nutritionally related problems, as well as changes in the local and national infrastructure of nutrition support services, are required to improve the quality of care and the clinical outcome for patients being treated by parenteral and enteral nutrition in hospital and at home.


Super resolution methodology based on temperature dependent Raman scattering.
The recent advances in far-field super-resolution (SR) microscopy rely on, and therefore are limited by the ability to control the fluorescence of label molecules. We suggest a new, label-free, far-field SR microscopy based on temperature dependence of Raman scattering. Here, we present simulation and experimental characterization of the method. In an ultrafast pump-probe scheme, a spatial temperature profile is optically excited throughout the diffraction-limited spot; the Raman spectrum is probed with an overlapping laser. Thermally induced shifts, recorded in a specific spectral region of interest (ROI), enable spatial discrimination between areas of different temperature. Our simulations show spatial resolution that surpasses the diffraction limit by more than a factor of 2. Our method is compatible with material characterization in ambient, vacuum and liquid, thin and thick samples alike.


Ca2+ imaging in plants using genetically encoded Yellow Cameleon Ca2+ indicators.
Temporally and spatially defined changes in cellular calcium (Ca(2+)) concentration represent stimulus-specific signals and regulate a myriad of biological processes. The development of ratiometric Ca(2+) reporter proteins like Yellow Cameleons (YCs) has greatly advanced our ability to analyze Ca(2+) dynamics in vivo with unprecedented spatial and temporal resolution. In plants, the application of these Ca(2+) reporter proteins has been pioneered for the investigation of Ca(2+) dynamics in guard cells, and recently their use has been extended to other single-cell models like growing pollen tubes and root hairs. However, in plants, the use of YC reporter proteins has largely remained restricted to the investigation of cytoplasmic alterations of Ca(2+) concentrations. Here, we provide an introduction to current methods for imaging Ca(2+) dynamics with increasing sophistication.


A simple protocol to characterize bacterial cell-envelope lipoproteins in a native-like environment.
Physiological conditions in living cells are strictly regulated to allow, optimize, and coordinate biological processes. The bacterial cell envelope is the compartment where the communication with the external environment takes place. This involves membrane proteins, key players in many biological processes that ensure bacterial survival. The biochemical characterization of membrane proteins, either integral, lipidated or peripheral is challenging due to their mixed protein-lipid nature, making it difficult to purify and obtain considerable amounts of samples. In contrast to integral membrane proteins, lipidated proteins are usually purified as truncated soluble versions, neglecting the impact of the membrane environment. Here we report a simple and robust protocol to characterize bacterial lipidated proteins in spheroplasts from Escherichia coli using a β-lactamase as a model. The Metallo-β-lactamase NDM-1 is an enzyme anchored to the inner leaflet of the outer membrane of Gram-negative bacteria. Kinetic parameters and stability of the lipidated NDM-1 and the soluble unbound version (NDM-1 C26A) were measured in spheroplasts and periplasm, respectively. These studies revealed that membrane anchoring increases the KM of the enzyme, consequently decreasing the catalytic efficiency, while not affecting its kinetic stability. This approach can be used to characterize lipidated proteins avoiding the purification step while mimicking its native environment. This approach also helps in filling the gap between in vitro and in vivo studies.


Heat-Inactivation of Streptococcus faecium var. casseliflavus in Skim Milk Cultures with Pseudomonas fluorescens.
Thermal destruction of Streptococcus faecium var. casseliflavus (SFC) at 57°C in autoclaved skim milk was determined at several pH values, and when skim milk was inoculated with Pseudomonas fluorescens and incubated 3 d at 10°C plus 4 d at room temperature before SFC was added. The pH values between 6.4 and 6.6 had little effect on heat resistance of SFC. At pH 5.6, however, accelerated thermal destruction occurred in sterile skim milk as compared to skim milk at pH 6.5. (D-values were 4.8 min and 10.2 min, respectively). Presence of large populations (log10 bacterial count = 9.8 to 9.9/ml) of P. fluorescens had a protective effect on SFC (D = 7.6 ± .7 min in skim milk preincubated with P. fluorescens and 6.2 min in skim milk without P. fluorescens ).


Fusion of the green fluorescent protein to amino acids 1 to 71 of bovine respiratory syncytial virus glycoprotein G directs the hybrid polypeptide as a class II membrane protein into the envelope of recombinant bovine herpesvirus-1.
It was recently shown that the class II membrane glycoprotein G of bovine respiratory syncytial virus (BRSV) is integrated into the envelope of recombinant bovine herpesvirus-1 (BHV-1) virions in the correct orientation. To verify the hypothesis that the membrane anchor of BRSV G might be suitable to target heterologous polypeptides into the membrane of recombinant BHV-1 particles, an open reading frame encoding a fusion protein between amino acids 1 to 71 of the BRSV G glycoprotein and the green fluorescent protein (TMIIGFP) was recombined into the genome of BHV-1. The resulting recombinant BHV-1/eTMIIGFP had growth properties similar to those of wild-type BHV-1. Live-cell analysis of cells infected with BHV-1/eTMIIGFP indicated that the fusion protein localized to the cell surface. Immunoprecipitations and virus neutralization assays using a GFP-specific antiserum proved that TMIIGFP was incorporated as a class II membrane protein into virions.


A midgut lysate of the Riptortus pedestris has antibacterial activity against LPS O-antigen-deficient Burkholderia mutants.
Riptortus pedestris, a common pest in soybean fields, harbors a symbiont Burkholderia in a specialized posterior midgut region of insects. Every generation of second nymphs acquires new Burkholderia cells from the environment. We compared in vitro cultured Burkholderia with newly in vivo colonized Burkholderia in the host midgut using biochemical approaches. The bacterial cell envelope of in vitro cultured and in vivo Burkholderia differed in structure, as in vivo bacteria lacked lipopolysaccharide (LPS) O-antigen. The LPS O-antigen deficient bacteria had a reduced colonization rate in the host midgut compared with that of the wild-type Burkholderia. To determine why LPS O-antigen-deficient bacteria are less able to colonize the host midgut, we examined in vitro survival rates of three LPS O-antigen-deficient Burkholderia mutants and lysates of five different midgut regions. The LPS O-antigen-deficient mutants were highly susceptible when cultured with the lysate of a specific first midgut region (M1), indicating that the M1 lysate contains unidentified substance(s) capable of killing LPS O-antigen-deficient mutants. We identified a 17 kDa protein from the M1 lysate, which was enriched in the active fractions. The N-terminal sequence of the protein was determined to be a soybean Kunitz-type trypsin inhibitor. These data suggest that the 17 kDa protein, which was originated from a main soybean source of the R. pedestris host, has antibacterial activity against the LPS O-antigen deficient (rough-type) Burkholderia.


Enhanced translation of heme oxygenase-2 preserves human endothelial cell viability during hypoxia.
Heme oxygenases (HOs) -1 and -2 catalyze the breakdown of heme to release carbon monoxide, biliverdin, and ferrous iron, which may preserve cell function during oxidative stress. HO-1 levels decrease in endothelial cells exposed to hypoxia, whereas the effect of hypoxia on HO-2 expression is unknown. The current study was carried out to determine if hypoxia alters HO-2 protein levels in human endothelial cells and whether this enzyme plays a role in preserving their viability during hypoxic stress. Human umbilical vein endothelial cells (HUVECs), human aortic endothelial cells (HAECs), and human blood outgrowth endothelial cells were exposed to 21% or 1% O(2) for 48 or 16 h in the presence or absence of tumor necrosis factor-alpha (10 ng/ml) or H(2)O(2) (100 microm). In all three endothelial cell types HO-1 mRNA and protein levels were decreased following hypoxic incubation, whereas HO-2 protein levels were unaltered. In HUVECs HO-2 levels were maintained during hypoxia despite a 57% reduction in steady-state HO-2 mRNA level and a 43% reduction in total protein synthesis. Polysome profiling revealed increased HO-2 transcript association with polysomes during hypoxia consistent with enhanced translation of these transcripts. Importantly, inhibition of HO-2 expression by small interference RNA increased oxidative stress, exacerbated mitochondrial membrane depolarization, and enhanced caspase activation and apoptotic cell death in cells incubated under hypoxic but not normoxic conditions. These data indicate that HO-2 is important in maintaining endothelial viability and may preserve local regulation of vascular tone, thrombosis, and inflammatory responses during reductions in systemic oxygen delivery.


Adoptive immunotherapy of solid tumors with activated macrophages: experimental and clinical results.
Adoptive immunotherapy in cancer has been essentially restricted to the use of lymphoid effector cells (NK, TIL, LAK) stimulated with IL-2. Differentiated macrophages represent another key effector population even more important for the immune control of cancer. We have shown that activated murine macrophages reduced primary tumors and experimental metastases. Human macrophages differentiated from circulating monocytes and activated with IFN gamma (MAK) were cytotoxic in vitro for a variety of tumor cell and caused regression of human tumors implanted in nude mice. A large scale technology has been developed for the generation of antitumor macrophages. These MAK cells (10(8) to 10(9] were injected in cancer patients in pilot clinical trials and were well tolerated. MAK treatment is technically feasible, clinically safe and presents several advantages compared to other immunotherapies.


Keratinocyte growth factor, tumor necrosis factor-alpha and interleukin-1 beta gene expression in cultured fibroblasts and keratinocytes from burned patients.
To evaluate the gene expression of KGF, TNF-alpha and IL-1 beta in skin fibroblasts and keratinocytes cultured from burned patients. Three patients with large burns and three patients with small burns, as well as two controls, were included. The cell culture was initiated by the enzymatic method. After extraction and purification of mRNA, qPCR was used to assess the gene expression of KGF, TNF-alpha and IL-1 beta. The expression of KGF was increased on average 220-fold in large burns and 33.33-fold in small burns in fibroblasts, and 11.2-fold in large burns and 3.45-fold in small burns in keratinocytes compared to healthy patients (p<0.05). Expression of TNF-alpha was not observed. IL-1 beta is down-regulated in fibroblasts of burned patients, and much more repressed in small burns (687-fold, p<0.05). In keratinocytes, the repression of IL-1 beta expression occurs in patients with small burns (28-fold), while patients with large burns express this gene intensively (15-fold). The study showed a quantitative pattern in the expression of KGF gene, which is more expressed according to the size of the burn. TNF-alpha was not expressed. A qualitative pattern in the expression of IL-1 beta gene was demonstrated.


[Squamous carcinoma of the prostate].
A primary squamous cell carcinoma occurring in the prostate of 84 year-old man is described. The patient died of local progression six months after diagnosis. Review of the literature suggests that such a cancer of the prostate is rare, highly aggressive, and responds poorly to any mode of therapy.


Prediction of fracture risk in postmenopausal white women with peripheral bone densitometry: evidence from the National Osteoporosis Risk Assessment.
Low bone mineral density (BMD) is a risk factor for fracture. Although the current "gold standard" test is DXA of the hip and spine, this method is not universally available. No large studies have evaluated the ability of new, less expensive peripheral technologies to predict fracture. We studied the association between BMD measurements at peripheral sites and subsequent fracture risk at the hip, wrist/forearm, spine, and rib in 149,524 postmenopausal white women, without prior diagnosis of osteoporosis. At enrollment, each participant completed a risk assessment questionnaire and had BMD testing at the heel, forearm, or finger. Main outcomes were new fractures of the hip, wrist/forearm, spine, or rib within the first 12 months after testing. After 1 year, 2259 women reported 2340 new fractures. Based on manufacturers' normative data and multivariable adjusted analyses, women who had T scores < or = -2.5 SD were 2.15 (finger) to 3.94 (heel ultrasound [US]) times more likely to fracture than women with normal BMD. All measurement sites/devices predicted fracture equally well, and risk prediction was similar whether calculated from the manufacturers' young normal values (T scores) or using SDs from the mean age of the National Osteoporosis Risk Assessment (NORA) population. The areas under receiver operating characteristic (ROC) curves for hip fracture were comparable with those published using measurements at hip sites. We conclude that low BMD found by peripheral technologies, regardless of the site measured, is associated with at least a twofold increased risk of fracture within 1 year, even at skeletal sites other than the one measured.