GLI1 is involved in cell cycle regulation and proliferation of NT2 embryonal carcinoma stem cells.
Hedgehog (HH) signaling plays a critical role during embryogenesis and regulates early development of multiple tissues and organs, including the central nervous system. Although much has been revealed of the diverse functions of the HH signaling pathway, it is still unclear how the effects of altered HH signaling are interpreted by specific cell types. We have investigated the role of the HH transcription factor glioma-associated oncogene homolog 1 (GLI1) in the human Ntera2/D1 (NT2) embryonal carcinoma stem cell line. The study revealed that expression of GLI1 and its direct transcriptional target Patched (PTCH) is downregulated in the early stages of retinoic acid-induced neuronal differentiation of NT2 cells. To identify transcriptional targets of the HH transcription factor GLI1 in NT2 cells, we performed global expression profiling following GLI1 RNA interference (RNAi). Of the 8500 transcripts represented on the microarrays, expression of 88 genes was downregulated and expression of 26 genes was upregulated. Nineteen of these genes are involved in cell cycle and proliferation. Further, GLI1 RNAi leads to a significant decrease in NT2 proliferation and changes expression of G1 phase cyclins. In conclusion, our results suggest that GLI1 is involved in cell cycle and proliferation control in the embryonal carcinoma stem cell line NT2.


[Knowledge in nursing: from the area representation to the Nursing Advisory Committee at CNPq].
The aim was to describe aspects of the nursing evolution in the National Council for Scientific and Technological Development (Conselho Nacional de Desenvolvimento Científico e Tecnológico - CNPq), focusing on the organizational structure and positions of the Area Representation and the advances in nursing knowledge. This is an experience report, accompanied by reflections and attitudes towards science, technology and innovation of Brazilian nursing and the creation of the Nursing Advisory Committee at the CNPq, in 2006. This paper is intended for the special issue of the Brazilian Nursing Journal (Revista Brasileira de Enfermagem), a prominent scientific nursing journal of nursing, in the celebration of its 80 years of existence. Brazilian nursing records scientific qualification advances in the preparation of its researchers, marking a new era in the consolidation and recognition of the discipline and profession.


Serum levels of astroglial S100-beta and neuron-specific enolase in hepatic encephalopathy patients.
To find a reliable, noninvasive method for the diagnosis of cognitive impairment in patients with hepatic cirrhosis we measured serum levels of astroglial S100beta and neuron-specific enolase in cirrhotic patients with and without hepatic encephalopathy (HE). S100beta levels showed a significant increase in groups with HE stage 1 and 2 compared to both control and cirrhosis patients. However serum neuron-specific enolase levels were not significantly different between the studied groups. S100beta levels had a specificity of 91.3% and sensitivity of 51.7% for detection of HE from cirrhosis. Serum S100beta may be a useful surrogate marker for the diagnosis of mild cognitive impairment in cirrhotic patients before they progress to more advanced stages of HE.


[Repeat operations on the biliary tract in a district hospital].
An experience with 79 reoperations in 68 patients who had been subjected to operations for acute and chronic cholecystitis enabled the author to conclude that retained calculi in bile ducts were the main cause of reoperations. The amount of repeated surgical interventions and postoperative lethality can be decreased by a complex intraoperative examination of bile ducts during the first operation and by higher qualification of the surgeons.


Elementary steps in the reaction mechanism of chicken liver fatty acid synthase: reduced nicotinamide adenine dinucleotide phosphate binding and formation and reduction of acetoacetyl-enzyme.
The kinetics of reduced nicotinamide adenine dinucleotide phosphate (NADPH) binding to fatty acid synthase from chicken liver and of the reduction of enzyme-bound acetoacetyl by NADPH (beta-ketoacyl reductase) and the steps leading to formation of the acetoacetyl-enzyme have been studied in 0.1 M potassium phosphate-1 mM ethylenediaminetetraacetic acid (EDTA), pH 7.0, at 25 degrees C by monitoring changes in NADPH fluorescence with a stopped-flow apparatus. Improved fluorescence detection has permitted the use of NADPH concentrations as low as 20 nM. The kinetics of the binding of NADPH to the enzyme is consistent with a simple bimolecular binding mechanism and four equivalent sites on the enzyme (presumably two beta-ketoacyl reductase sites and two enoyl reductase sites). The bimolecular rate constant is 12.7 X 10(6) M-1 s-1, and the dissociation rate constant is 76.7 s-1, which gives an equilibrium dissociation constant of 6.0 microM. The formation of the acetoacetyl-enzyme and its subsequent reduction by NADPH could be analyzed as two consecutive pseudo-first-order reactions by mixing enzyme-NADPH with acetyl-CoA and malonyl-CoA under conditions where [acetyl-CoA], [malonyl-CoA] much greater than [enzyme] much greater than [NADPH]. From the dependence of the rate of reduction of aceto-acetyl-enzyme by NADPH on enzyme concentration, an independent estimate of the equilibrium dissociation constant for NADPH binding to the enzyme of 5.9 microM is obtained, and the rate constant for the reduction is 17.5 s-1.(ABSTRACT TRUNCATED AT 250 WORDS)


Three Diagnoses for Problematic Hypersexuality; Which Criteria Predict Help-Seeking Behavior?
This study aimed to assess the best combination of indicators of problematic hypersexuality (PH), in a survey (n = 58,158) targeting individuals wondering if they were sex addicted. The survey allowed for testing of criteria from three theoretical models used to conceptualize PH. Factor analyses for women and men yielded an interpretable grouping of indicators consisting of four factors. In subsequent logistic regressions, these factors were used as predictors for experiencing the need for help for PH. The factors Negative Effects and Extreme positively predicted experiencing the need for help, with Negative Effects as the most important predictor for both women and men. This factor included, among others, withdrawal symptoms and loss of pleasure. The Sexual Desire factor negatively predicted the need for help, suggesting that for the targeted population more sexual desire leads to less PH. The Coping factor did not predict experiencing the need for help. Outcomes show that a combination of indicators from different theoretical models best indicates the presence of PH. Therefore, a measurement instrument to assess existence and severity of PH should consist of such a combination. Theoretically, this study suggests that a more comprehensive model for PH is needed, surpassing existing conceptualizations of PH.


The swarming motility of Pseudomonas aeruginosa is blocked by cranberry proanthocyanidins and other tannin-containing materials.
Bacterial motility plays a key role in the colonization of surfaces by bacteria and the subsequent formation of resistant communities of bacteria called biofilms. Derivatives of cranberry fruit, predominantly condensed tannins called proanthocyanidins (PACs) have been reported to interfere with bacterial adhesion, but the effects of PACs and other tannins on bacterial motilities remain largely unknown. In this study, we investigated whether cranberry PAC (CPAC) and the hydrolyzable tannin in pomegranate (PG; punicalagin) affected the levels of motilities exhibited by the bacterium Pseudomonas aeruginosa. This bacterium utilizes flagellum-mediated swimming motility to approach a surface, attaches, and then further spreads via the surface-associated motilities designated swarming and twitching, mediated by multiple flagella and type IV pili, respectively. Under the conditions tested, both CPAC and PG completely blocked swarming motility but did not block swimming or twitching motilities. Other cranberry-containing materials and extracts of green tea (also rich in tannins) were also able to block or impair swarming motility. Moreover, swarming bacteria were repelled by filter paper discs impregnated with many tannin-containing materials. Growth experiments demonstrated that the majority of these compounds did not impair bacterial growth. When CPAC- or PG-containing medium was supplemented with surfactant (rhamnolipid), swarming motility was partially restored, suggesting that the effective tannins are in part acting by a rhamnolipid-related mechanism. Further support for this theory was provided by demonstrating that the agar surrounding tannin-induced nonswarming bacteria was considerably less hydrophilic than the agar area surrounding swarming bacteria. This is the first study to show that natural compounds containing tannins are able to block P. aeruginosa swarming motility and that swarming bacteria are repelled by such compounds.


Is History of Periodontitis a Risk Factor for Peri-implant Disease? A Pilot Study.
The aim of this pilot study was to evaluate clinical outcomes of implant treatment in periodontally compromised patients. Partially edentulous patients who were diagnosed and treated for chronic periodontitis and later rehabilitated with implant-supported single crowns or fixed partial dentures were enrolled in this study. At the final follow-up, data on probing pocket depth and bleeding on probing were collected, and changes in peri-implant bone levels were assessed on periapical radiographs. Accordingly, patients were categorized into peri-implant disease-free (PID-free), peri-implant mucositis (PIM), or peri-implantitis (PI) groups. Additionally, information on the history of systemic diseases was obtained and correlated with the findings. Seven females and six males with a mean age of 55.2 (standard deviation [SD] ± 8.38) years were included in this study. Fifty-five implants were placed in 13 partially edentulous patients with a history of chronic periodontitis. The mean follow-up in the study was 35.1 (SD ± 2.40) months. No implants were lost in the 13 patients, for a cumulative survival of 100%. PI was confirmed in 3 patients and PIM in 8 patients, while 2 patients were found to be PID-free. Diabetes and hypertension were found to be the predominant systemic factors among patients affected by peri-implant diseases. Patients with a history of periodontitis are at considerable risk of being affected by peri-implant mucositis, if not by peri-implantitis.


CFTR: a hub for kinases and crosstalk of cAMP and Ca2+.
Cystic fibrosis (CF) is caused by mutations in the CF transmembrane conductance regulator (CFTR). The resulting disease is pleiotropic consistent with the idea that CFTR acts as a node within a network of signalling proteins. CFTR is not only a regulator of multiple transport proteins and controlled by numerous kinases but also participates in many signalling pathways that are disrupted after expression of its commonest mutant (F508del-CFTR). It operates in membrane compartments creating a scaffold for cytoskeletal elements, surface receptors, kinases and phosphodiesterases. CFTR is exposed to membrane-local second messengers such that a CFTR-interacting, low cellular energy sensor kinase (AMP- and ADP-activated kinase, AMPK) signals through a high energy phosphohistidine protein kinase (nucleoside diphosphate kinase, NDPK). CFTR also translocates a Ca(2+)-dependent adenylate cyclase to its proximity so that a rigid separation between cAMP-dependent and Ca(2+)-dependent regulation of Cl(-) transport becomes obsolete. In the presence of wild-type CFTR, parallel activation of CFTR and outwardly rectifying anoctamin 6 Cl(-) channels is observed, while the Ca(2+)-activated anoctamin 1 Cl(-) channel is inhibited. In contrast, in CF cells, CFTR is missing/mislocalized and the outwardly rectifying chloride channel is attenuated while Ca(2+)-dependent Cl(-) secretion (anoctamin 1) appears upregulated. Additionally, we consider the idea that F508del-CFTR when trapped in the endoplasmic reticulum augments IP3-mediated Ca(2+) release by providing a shunt pathway for Cl(-). CFTR and the IP3 receptor share the characteristic that they both assemble their partner proteins to increase the plasticity of their hub responses. In CF, the CFTR hub fails to form at the plasma membrane, with widespread detrimental consequences for cell signalling.


[Functional condition of temporal and masticatory muscles in patients with malocclusion complicated by crowding of frontal teeth].
To study functional condition of temporal and masticatory muscles in patients with malocclusion, complicated by crowding of frontal teeth, 54 patients were subjected to electromyography. It was found that the degree of functional imbalance of masticatory and temporal muscles depends on the type of crowding of frontal teeth. Investigating phases of physiological rest in patients with crowding of maxilla frontal teeth high indices of oscillation amplitude of temporal muscles biopotentials (P < 0,05) were detected. The study has established that in state of compression in patients with crowding of frontal teeth of maxilla and mandibula temporal muscles are more active, what is connected with insufficient activity of masticatory muscles (P < 0,05).


Effect of booster seat laws on appropriate restraint use by children 4 to 7 years old involved in crashes.
To quantify the independent contribution of recently enacted booster seat laws on appropriate restraint use by child passengers in motor vehicles. Longitudinal study of children involved in crashes with data collected via insurance claims records and a validated telephone survey. Sixteen states and Washington, DC, from December 1, 1998, through December 31, 2004. Probability sample of 5198 vehicles in crashes involving 6102 children aged 4 to 7 years, representing 78 159 vehicles and 91 752 children. Booster seat law provisions, child age, state, and secular trends. Reported appropriate restraint use for this age group, including forward-facing child safety seats, belt-positioning booster seats, and combination seats. Children aged 4 to 7 years in states with booster seat laws were 39% more likely to be reported as appropriately restrained than were children in other states (prevalence ratio [PR], 1.39; 95% confidence interval [CI], 1.14-1.70). Children aged 4 to 5 years were 23% more likely (PR,1.23; 95% CI, 0.80-1.42) and children 6 to 7 years twice as likely (PR, 2.09; 95% CI, 1.46-2.99) to be reported as appropriately restrained. For children aged 6 to 7 years, when compared with no law, laws through age 7 years were most effective (PR, 3.71; 95% CI, 2.49-5.42), followed by laws through age 4 or 5 years (PR, 1.43; 95% CI, 0.89-2.24). Given the higher current use of age-appropriate restraints among children 4 to 5 years compared with older children, future upgrades to child restraint laws should include children through at least age 7 years to maximize the number of children properly restrained for their age.


Safety of percutaneous dilatational tracheostomy in patients on extracorporeal lung support.
To evaluate the safety of percutaneous dilatational tracheostomy (PDT) in critically ill patients on an extracorporeal lung assist device requiring therapeutic anticoagulation. This was a retrospective, observational study on all patients undergoing tracheostomy while on pumpless extracorporeal lung assist or extracorporeal membrane oxygenation in intensive care units of two university hospitals in Germany between 2007 and 2013. During the study period PDT was performed on 118 patients. The median platelet count, international normalized ratio, and activated partial thromboplastin time before tracheostomy were 126 × 10(9)/L (range 16-617 × 10(9)/L), 1.1 (0.9-2.0) and 49 s (28-117 s), respectively. Seventeen patients (14.4%) received a maximum of three bags of pooled platelets, and eight patients (6.8%) received a maximum of four units of fresh frozen plasma before the procedure. In all patients the administration of intravenous heparin was briefly paused periprocedurally. No periprocedural clotting complication within the extracorporeal circuit was observed. Two patients (1.7%) suffered from procedure-related major bleeding, with one patient requiring conversion to a surgical tracheostomy. Two pneumothoraces (1.7%) were related to the PDT. One patient (0.8%) had analgosedation-related hypotension with brief and successful cardiopulmonary resuscitation. Minor bleeding from the tracheostomy site occurred in 37 cases (31.4%). No fatality was attributable to tracheostomy. The complication rates of PDT in the patients on extracorporeal lung support were low and comparable to those of other critically ill patients. Based on these results, we conclude that PDT performed by experienced operators with careful optimization of the coagulation state is a relatively safe procedure and not contraindicated for this patient group.


Rumen morphometrics and the effect of digesta pH and volume on volatile fatty acid absorption.
The effects of rumen digesta volume and pH on VFA absorption and its relation to rumen wall morphology were evaluated. Nine rumen cannulated cows formed 3 groups based on desired variation in rumen morphology: The High group was formed by Holsteins yielding 25.9 kg milk/d and fed on a high-grain total mixed ration (TMR); the Medium group by Holstein-Zebu crossbreds yielding 12.3 kg milk/d and fed on corn silage, tropical pasture, and a commercial concentrate; and the Dry group by nonlactating grazing Jerseys fed exclusively on tropical pasture. Within each group, a sequence of 3 ruminal conditions was induced on each cow in 3 × 3 Latin Squares, with 7-d periods: high digesta volume and high pH (HVHP), low volume and high pH (LVHP), and low volume and low pH (LVLP). Rumen mucosa was biopsied on the first day of Period 1. Ruminal morphometric variables evaluated were mitotic index, absorptive surface and papillae number per square centimeter of wall, area per papillae, papillae area as a percentage of absorptive surface, and epithelium, keratinized layer, and nonkeratinized layer thickness. There was marked variation in rumen morphology among the groups of cows. Grazing Jerseys had decreased rumen wall absorptive surface area and basal cells mitotic index, and increased thickness of the epithelium and of the keratin layer compared with cows receiving concentrates. Mean rumen pH throughout the 4 h sampling period was: 6.78 for HVHP, 7.08 for LVHP, and 5.90 for LVLP (P < 0.01). The capacity of the rumen wall to absorb VFA was estimated by the Valerate/CrEDTA technique. The fractional exponential decay rate for the ratio of valeric acid to Cr (k Val/Cr) was determined by rumen digesta sampling at 20-min intervals during 4 h, after the mixing of markers and the return of the evacuated ruminal content. The k Val/Cr values for treatments HVHP, LVHP, and LVLP were, respectively: 19.6, 23.9, and 35.0 %/h (SEM = 2.01; P = 0.21 for contrast HVHP vs. LVHP and P < 0.01 for contrast LVHP vs. LVLP). The k Val/Cr was faster under low pH, but decreasing digesta volume under high pH did not elicit such a response. The correlation between the absorptive surface area per square centimeter of rumen wall and the mean of the 3 k Val/Cr values of each cow was 0.90 (P < 0.01). Cows capable of maintaining a less-acidic rumen environment had greater inflow of water into the digestive cavity, had a more developed rumen mucosa, and were more efficient VFA absorbers.


Association between VDR genetic polymorphisms and risk of gestational diabetes mellitus in the Chinese population.
Abnormal metabolism of vitamin D was the primary mechanism in many pregnancy diseases. Our study was the first to examine the hypothesis that VDR gene polymorphisms contribute to the risk of gestational diabetes mellitus (GDM) in the Chinese population at high altitudes. One hundred and eighteen women with GDM and 104 women with normal glucose tolerance (NGT) were included in this study using a case-control design. Four single nucleotide polymorphisms (g.47879112G > A, g.47846052C > T, g.47844974A > G, and g.47845054C > A) of mother and fetus were genotyped. Maternal and fetal frequency of the A allele of g.47879112G > A was significantly increased in women with GDM than in those with NGT (p < .05). A correlation between the AA homozygous genotype of g.47879112G > A and GDM was noted. Compared with non-carriers, A allele carriers showed higher fasting plasma insulin and two-hour post-challenge plasma glucose (2h-PPG), and lower levels of vitamin D. Furthermore, both maternal and fetal 4-marker haplotype ACCG were found to be significantly associated with GDM (p < .05). Association and haplotype analysis indicated that the A allele of g.47879112G > A could be a risk factor for GDM development in the Chinese population at high altitudes. Additionally, the VDR gene polymorphism of the fetus and mother may have a synergistic effect. The VDR polymorphism is associated with an increased risk of GDM and may be useful for predicting the development of the disease.


Why do people with complex regional pain syndrome take longer to recognize their affected hand?
People with complex regional pain syndrome (CRPS) take longer to recognize the laterality of a pictured hand when it coincides with their affected hand. The author explored two aspects of this phenomenon: whether the duration of symptoms relates to the extent of the delay and whether guarding-type mechanisms are involved. Eighteen patients with CRPS type 1 of the wrist and 18 matched control subjects performed a hand laterality recognition task. McGill pain questionnaire, Neuropathic Pain Scale, and response time (RT) to recognize hand laterality were analyzed. Regressions related 1) mean RT for patients to the duration of symptoms and to pain intensity; and 2) mean RT for each picture to the predicted pain on executing that movement as judged by the patient, and to the awkwardness of the movement that would be required. For patients, the duration of symptoms correlated with mean RT (Spearman rho = 0.44; p = 0.02). Predicted pain rating explained 45% of the variance in RT for each picture for each patient (p < 0.01). The results suggest that in patients with complex regional pain syndrome type 1, delayed recognition of hand laterality is related to the duration of symptoms and to the pain that would be evoked by executing the movement. The former is consistent with chronic pain and disuse and may involve reorganization of the cortical correlate of body schema. The latter is consistent with a guarding-type response that probably occurs upstream of the motor cortex at a motor planning level.


[Pediatric femoral shaft fractures. Evaluation of 504 follow-up studies].
From 657 children who suffered from fractures of the femoral shaft during the years 1965-1980 504 could be followed-up in 13 hospitals of the GDR clinically and roentgenologically. 313 had undergone conservative treatment, 191 surgery with different methods. In all surgically treated cases there was overgrowth of the broken femoral shaft there was a shortening only in a few cases. In most cases the rotational deformity of the distal fragment persisted. Dislocation ad latus always had disappeared, whereas varus and expecially valgus deformities as well as antecurvation had improved partially. Coxa valga deformity was found most frequently after internal fixation.


What do adult squash players think about protective eyewear?
To determine the attitudes of adult squash players towards protective eyewear. A survey of 197 competition and social squash players from seven squash centres in the outer eastern suburbs of Melbourne, Australia was conducted in September to October 1995. Information about participation in squash, previous injuries, use of protective eyewear, barriers towards eyewear use, and attitudes towards protective eyewear was obtained by a self report questionnaire. Squash is a popular sport in Australia. Of the players surveyed, 6% played in junior competitions, 67% in senior competitions, and 27% were social players. Most had been playing for more than ten years. Some 15% of players had previously suffered an eye injury, most commonly caused by a racquet. Less than 10% of players reported that they wore protective eyewear when they played squash, and 35% of these wore prescriptive lenses which they considered to be protective. The major reason for not wearing protective eyewear was the perception that it was unnecessary. Poor vision and a lack of comfort were also stated as reasons by a significant number of players. More than half (57%) of the respondents agreed that more players should wear protective eyewear, yet only 16% thought it should be compulsory for all players. There was considerable support for protective eyewear use by junior players, however. The rate of protective eyewear use is low among competition and social squash players in Melbourne. The major areas that need to be addressed are the ignorance of the need for protective eyewear among social and experienced players and the mistaken belief that prescription lenses provide adequate protection on a squash court.


Hemodynamic effects of a new right ventricular assist device.
A right ventricular assist device (VAD) based on the principle of counterpulsation has been developed at our institution. The device is a valveless, pneumatically actuated, 40 cc, sac-type pump, with a single inlet-outlet port. For right ventricular support, the "Uniport" pump is anastamosed end-to-side to the pulmonary artery. In previous experimental trials, the device has been shown to impart minimal trauma to blood components. In this study, biventricular failure was induced in eight Holstein calves by normothermic ischemia during cardiopulmonary bypass. A Pierce-Donachy left VAD (LVAD) was used for left ventricular support following the ischemic insult. Hemodynamic measurements were obtained throughout the study, and each animal served as its own control. A significant increase in post injury cardiac output (33.5 +/- 11.4%) was obtained with use of the Uniport and LVAD, as compared to use of the LVAD alone (p less than or equal to 0.005). Other hemodynamic parameters of right heart failure, including right atrial pressure (RAP), pulmonary artery pressure (PAP), and left atrial pressure (LAP) were not significantly affected. These data suggest that the Uniport right ventricular assist device significantly improves cardiac output in this model of moderate right ventricular failure. Additional studies are required, however, to optimize pump stroke volume, and to further define the performance envelope of the device.


The expression and cellular localization of the sperm flagellar protein MC31/CE9 in the rat testis: possible posttranscriptional regulation during rat spermiogenesis.
We isolated the MC31 cDNA clone coding the antigen specifically recognized by the monoclonal antibody mMC31, and found that MC31 was identical to rat CE9. Therefore, this molecule is called MC31/CE9. MC31/CE9, a member of the immunoglobulin superfamily molecules, was localized on the rat sperm flagellar plasma membrane. We analyzed the expression and cellular localization of MC31/CE9 mRNA and protein in the adult rat testis by use of Northern hybridization, in situ hybridization, and immunohistochemical analyses. In the course of spermatogenesis, MC31/CE9 mRNA first appeared in type B spermatogonia. The mRNA signal intensity increased progressively to pachytene spermatocytes and remained constantly at a considerable level throughout the subsequent phases of spermatocytes and round spermatids, and then decreased gradually from step-11 spermatids to disappear in step-15 spermatids. On the other hand, MC31/CE9 protein expression showed a bimodal pattern. Immunohistochemical analysis for the MC31/CE9 protein revealed its most intense immunoreactivity on the flagella of step-8 to step-19 elongated spermatids. The cytoplasmic immunoreactivity of the MC31/CE9 protein also appeared in preleptotene to early pachytene spermatocytes and elongated spermatids, with particularly intense immunoreactivity in the Golgi complexes of zygotene and early pachytene spermatocytes (stage XIII to III) as well as step-8 to step-13 spermatids. Between these two phases, the MC31/CE9 protein proved undetectable in the cytoplasm of any spermatogenic cells. Sertoli cells and Leydig cells were devoid of MC31/CE9 mRNA and its protein. Therefore, the production of MC31/CE9 is thought to be posttranscriptionally regulated during spermiogenesis.


Xylose reductase from the thermophilic fungus Talaromyces emersonii: cloning and heterologous expression of the native gene (Texr) and a double mutant (TexrK271R + N273D) with altered coenzyme specificity.
Xylose reductase is involved in the first step of the fungal pentose catabolic pathway. The gene encoding xylose reductase (Texr) was isolated from the thermophilic fungus Talaromyces emersonii, expressed in Escherichia coli and purified to homogeneity. Texr encodes a 320 amino acid protein with a molecular weight of 36 kDa, which exhibited high sequence identity with other xylose reductase sequences and was shown to be a member of the aldoketoreductase (AKR) superfamily with a preference for reduced nicotinamide adenine dinucleotide phosphate (NADPH) as coenzyme. Given the potential application of xylose reductase enzymes that preferentially utilize the reduced form of nicotinamide adenine dinucleotide (NADH) rather than NADPH in the fermentation of five carbon sugars by genetically engineered microorganisms, the coenzyme selectivity of TeXR was altered by site-directed mutagenesis. The TeXR K271R+N273D double mutant displayed an altered coenzyme preference with a 16-fold improvement in NADH utilization relative to the wild type and therefore has the potential to reduce redox imbalance of xylose fermentation in recombinant S. cerevisiae strains. Expression of Texr was shown to be inducible by the same carbon sources responsible for the induction of genes encoding enzymes relevant to lignocellulose hydrolysis, suggesting a coordinated expression of intracellular and extracellular enzymes relevant to hydrolysis and metabolism of pentose sugars in T. emersonii in adaptation to its natural habitat. This indicates a potential advantage in survival and response to a nutrient-poor environment.


The release and induced immune responses of a plant-made and delivered antigen in the mouse gut.
This study investigated the site of release of a model vaccine antigen from plant cells and the corresponding induced immune response. Three plant tissues (leaf, fruit and hairy root) and two formulations (aqueous and lipid) were compared in two mouse trials. A developed technique that enabled detection of antigen release by plant cells determined that antigen release occurred at early sites of the gastrointestinal tract when delivered in leaf material and at later sites when delivered in hairy roots. Lipid formulations delayed antigen release from all plant materials tested. While encapsulation in the plant cell provided some protection of the antigen in the gastrointestinal tract and influenced antigen release, formulation medium was also an important consideration with regard to vaccine delivery and immunogenicity. Systemic immune responses induced from the orally delivered vaccine benefited from late release of antigen in the mouse gastrointestinal tract. The influences to the mucosal immune response induced by these vaccines were too complex to be determined by studies performed here with no clear trend regarding plant tissue site of release or formulation medium. Expression and delivery of the model antigen in plant material prepared in an aqueous formulation provided the optimal systemic and mucosal, antigen-specific immune responses.


Geographic determinants of reported human Campylobacter infections in Scotland.
Campylobacteriosis is the leading cause of bacterial gastroenteritis in most developed countries. People are exposed to infection from contaminated food and environmental sources. However, the translation of these exposures into infection in the human population remains incompletely understood. This relationship is further complicated by differences in the presentation of cases, their investigation, identification, and reporting; thus, the actual differences in risk must be considered alongside the artefactual differences. Data on 33,967 confirmed Campylobacter infections in mainland Scotland between 2000 and 2006 (inclusive) that were spatially referenced to the postcode sector level were analysed. Risk factors including the Carstairs index of social deprivation, the easting and northing of the centroid of the postcode sector, measures of livestock density by species and population density were tested in univariate screening using a non-spatial generalised linear model. The NHS Health Board of the case was included as a random effect in this final model. Subsequently, a spatial generalised linear mixed model (GLMM) was constructed and age-stratified sensitivity analysis was conducted on this model. The spatial GLMM included the protective effects of the Carstairs index (relative risk (RR) = 0.965, 95% Confidence intervals (CIs) = 0.959, 0.971) and population density (RR = 0.945, 95% CIs = 0.916, 0.974. Following stratification by age group, population density had a significant protective effect (RR = 0.745, 95% CIs = 0.700, 0.792) for those under 15 but not for those aged 15 and older (RR = 0.982, 95% CIs = 0.951, 1.014). Once these predictors have been taken into account three NHS Health Boards remain at significantly greater risk (Grampian, Highland and Tayside) and two at significantly lower risk (Argyll and Ayrshire and Arran). The less deprived and children living in rural areas are at the greatest risk of being reported as a case of Campylobacter infection. However, this analysis cannot differentiate between actual risk and heterogeneities in individual reporting behaviour; nevertheless this paper has demonstrated that it is possible to explain the pattern of reported Campylobacter infections using both social and environmental predictors.


Three-year results of an investigator-driven multicenter, international, randomized open-label de novo trial to prevent BOS after lung transplantation.
Chronic lung allograft dysfunction (CLAD), predominantly manifest as bronchiolitis obliterans syndrome (BOS), is the primary cause of morbidity and death after lung transplantation. We assessed the efficacy and safety of 2 de novo immunosuppression protocols to prevent BOS. This was a multicenter, prospective, international, randomized (1:1) open-label superiority study of de novo enteric-coated mycophenolate sodium (MPS) vs delayed-onset everolimus (RAD), both arms in combination with cyclosporine (CsA) monitored by 2-hour post-dose (C2) levels, and corticosteroids. Target C2 levels were lower in the RAD group because RAD is known to potentiate CsA nephrotoxicity. Cytolytic induction therapy was not used. Patients were stratified at entry for cystic fibrosis. Confirmation of anastomotic healing was required for randomization. Primary efficacy was freedom from BOS Grade 1 on intention-to-treat (ITT) analysis. Secondary efficacy parameters were patient and graft survival and severity of rejection. Treatment failure was defined by graft loss, patient death, drug cessation, or need for other therapy. The 3-year freedom from BOS Grade 1 was 70% for MPS (n = 80) vs 71% for RAD (n = 84; p = 0.95 by log-rank) in ITT but was lower in the RAD arm of the per-protocol population (p = 0.03). The 3-year survival was 84% (MPS) vs 76% (RAD; p = 0.19 by log-rank). Thirteen patients switched from MPS vs 31 from RAD (p < 0.01). Days on MPS were greater than days on RAD (p < 0.01). Rejection events proven by biopsy specimen were more common on MPS (p = 0.02), as were leucopenia (p < 0.01), diarrhea (p < 0.01), and cytomegalovirus infection (p = 0.04). Venous thromboembolism was more frequent on RAD (p = 0.02). Creatinine at 3 years was 160 ± 112 μmol/1iter in MPS patients vs 152 ± 98 μmol/1iter in RAD patients (p = 0.67). This 3-year ITT analysis found no significant difference between arms but was underpowered to accept the null hypothesis that RAD and MPS have equivalent efficacy in preventing BOS or death after lung transplantation.


The drift diffusion model as the choice rule in inter-temporal and risky choice: A case study in medial orbitofrontal cortex lesion patients and controls.
Sequential sampling models such as the drift diffusion model (DDM) have a long tradition in research on perceptual decision-making, but mounting evidence suggests that these models can account for response time (RT) distributions that arise during reinforcement learning and value-based decision-making. Building on this previous work, we implemented the DDM as the choice rule in inter-temporal choice (temporal discounting) and risky choice (probability discounting) using hierarchical Bayesian parameter estimation. We validated our approach in data from nine patients with focal lesions to the ventromedial prefrontal cortex / medial orbitofrontal cortex (vmPFC/mOFC) and nineteen age- and education-matched controls. Model comparison revealed that, for both tasks, the data were best accounted for by a variant of the drift diffusion model including a non-linear mapping from value-differences to trial-wise drift rates. Posterior predictive checks confirmed that this model provided a superior account of the relationship between value and RT. We then applied this modeling framework and 1) reproduced our previous results regarding temporal discounting in vmPFC/mOFC patients and 2) showed in a previously unpublished data set on risky choice that vmPFC/mOFC patients exhibit increased risk-taking relative to controls. Analyses of DDM parameters revealed that patients showed substantially increased non-decision times and reduced response caution during risky choice. In contrast, vmPFC/mOFC damage abolished neither scaling nor asymptote of the drift rate. Relatively intact value processing was also confirmed using DDM mixture models, which revealed that in both groups >98% of trials were better accounted for by a DDM with value modulation than by a null model without value modulation. Our results highlight that novel insights can be gained from applying sequential sampling models in studies of inter-temporal and risky decision-making in cognitive neuroscience.


Conversion of human steroid 5β-reductase (AKR1D1) into 3β-hydroxysteroid dehydrogenase by single point mutation E120H: example of perfect enzyme engineering.
Human aldo-keto reductase 1D1 (AKR1D1) and AKR1C enzymes are essential for bile acid biosynthesis and steroid hormone metabolism. AKR1D1 catalyzes the 5β-reduction of Δ(4)-3-ketosteroids, whereas AKR1C enzymes are hydroxysteroid dehydrogenases (HSDs). These enzymes share high sequence identity and catalyze 4-pro-(R)-hydride transfer from NADPH to an electrophilic carbon but differ in that one residue in the conserved AKR catalytic tetrad, His(120) (AKR1D1 numbering), is substituted by a glutamate in AKR1D1. We find that the AKR1D1 E120H mutant abolishes 5β-reductase activity and introduces HSD activity. However, the E120H mutant unexpectedly favors dihydrosteroids with the 5α-configuration and, unlike most of the AKR1C enzymes, shows a dominant stereochemical preference to act as a 3β-HSD as opposed to a 3α-HSD. The catalytic efficiency achieved for 3β-HSD activity is higher than that observed for any AKR to date. High resolution crystal structures of the E120H mutant in complex with epiandrosterone, 5β-dihydrotestosterone, and Δ(4)-androstene-3,17-dione elucidated the structural basis for this functional change. The glutamate-histidine substitution prevents a 3-ketosteroid from penetrating the active site so that hydride transfer is directed toward the C3 carbonyl group rather than the Δ(4)-double bond and confers 3β-HSD activity on the 5β-reductase. Structures indicate that stereospecificity of HSD activity is achieved because the steroid flips over to present its α-face to the A-face of NADPH. This is in contrast to the AKR1C enzymes, which can invert stereochemistry when the steroid swings across the binding pocket. These studies show how a single point mutation in AKR1D1 can introduce HSD activity with unexpected configurational and stereochemical preference.


Maintaining T lymphocyte homeostasis: another duty of autophagy.
First identified as a pathway for nutrient recovery during periods of starvation, the role of autophagy has expanded to the clearance of "toxic" intracellular material including ubiquitin-positive protein aggregates, damaged organelles as well as microbial pathogens in various cell types. We have examined the role of autophagy in the development and function of the adaptive immune system. Genes encoding autophagy machinery are expressed in T lymphocytes, and autophagy occurs in primary CD4+ and CD8+ T cells. By generating fetal liver chimeric mice, we found that thymocyte development is largely normal but the mature T cell compartment is severely reduced in the absence of the essential autophagy gene Atg5. Consistent with a critical role for autophagy in promoting T cell survival, Atg5-/- CD8+ T cells display high levels of apoptosis. Surprisingly, Atg5-deficient T cells were also unable to efficiently proliferate after T-cell receptor (TCR) stimulation. These findings suggest that autophagy regulates T lymphocyte homeostasis by promoting both survival and proliferation. In addition, T cells offer a new, physiologically relevant system to study the regulation and function of autophagy pathways in vivo.


Natural history of coronary artery disease in relation to angiographic, hemodynamic and clinical factors.
Studying the natural history of coronary artery disease could provide a frame of reference for prognosis and appraisal of treatment for patients having this disease. We studied a total of 465 consecutive patients with angiographically significant coronary artery disease, defined as greater than 50% stenosis in at least one principal artery, who were followed with medical treatment only from 1 to 7 years. Excluding patients with left main coronary disease, there were 73 deaths, of whom 63 were cardiac. The 5-year cumulative survival rates were 72% for the entire group, 87% for single vessel disease, 73% for double vessel disease and 51% for triple vessel disease. In single vessel disease, patients with left anterior descending artery involvement tended to have higher mortality. In double vessel disease, survival was worse with the combination of left anterior descending and right coronary artery involvements than the other 2 combinations. A history of myocardial infarction was not significantly different from angina in 5-year survival rate. Nevertheless, an abnormal Q-wave in ECG was associated with lower survival. History of hypertension and electrocardiographic left ventricular hypertrophy did not affect survival. While congestive heart failure, abnormal resting LVEDP and left ventricular asynergy were all associated with reduced survival. The left ventricular ejection fraction had highly prognostic value, only 42% of patients survived with ejection fraction less than 0.3 at the end of 5 years after angiography.


Case series of misdiagnosis with rK39 strip test in Indian leishmaniasis.
This report presents three cases where the rK39 strip test failed to diagnose two cases of post-kala-azar dermal leishmaniasis and one case of visceral leishmaniasis. However, a strong clinical suspicion prompted further evaluation by polymerase chain reaction (PCR), which established the etiology. The present case series highlights the usefulness of PCR in the diagnosis of leishmaniasis.


The Microbiome Meets Nanotechnology: Opportunities and Challenges in Developing New Diagnostic Devices.
Monitoring of the human microbiome is an emerging area of diagnostics for personalized medicine. Here, the potential of different nanomaterials and nanobiosensing technologies is reviewed for the development of novel diagnostic devices for the detection and measurement of microbiome-related biomarkers. Moreover, the current and future landscape of microbiome-based diagnostics is defined by exploring the advantages and disadvantages of current nanotechnology-based approaches, especially in the context of developing point-of-care (PoC) devices that would meet the international guidelines known as REASSURED (Real-time connectivity; Ease of specimen collection; Affordability; Sensitivity; Specificity; User-friendliness; Rapid & robust operation; Equipment-free; and Deliverability). Finally, the strategies of the latest international scientific consortia working in this field are analyzed, the current microbiome diagnostics market are reported and the principal ethical, legal, and societal issues related to microbiome R&D and innovation are discussed.


Cost-Effectiveness Thresholds in Global Health: Taking a Multisectoral Perspective.
Good health is a function of a range of biological, environmental, behavioral, and social factors. The consumption of quality health care services is therefore only a part of how good health is produced. Although few would argue with this, the economic framework used to allocate resources to optimize population health is applied in a way that constrains the analyst and the decision maker to health care services. This approach risks missing two critical issues: 1) multiple sectors contribute to health gain and 2) the goods and services produced by the health sector can have multiple benefits besides health. We illustrate how present cost-effectiveness thresholds could result in health losses, particularly when considering health-producing interventions in other sectors or public health interventions with multisectoral outcomes. We then propose a potentially more optimal second best approach, the so-called cofinancing approach, in which the health payer could redistribute part of its budget to other sectors, where specific nonhealth interventions achieved a health gain more efficiently than the health sector's marginal productivity (opportunity cost). Likewise, other sectors would determine how much to contribute toward such an intervention, given the current marginal productivity of their budgets. Further research is certainly required to test and validate different measurement approaches and to assess the efficiency gains from cofinancing after deducting the transaction costs that would come with such cross-sectoral coordination.


Novel mesostructured inclusions in the epidermal lining of Artemia franciscana ovisacs show optical activity.
Biomineralization, e.g., in sea urchins or mollusks, includes the assembly of mesoscopic superstructures from inorganic crystalline components and biopolymers. The resulting mesocrystals inspire biophysicists and material scientists alike, because of their extraordinary physical properties. Current efforts to replicate mesocrystal synthesis in vitro require understanding the principles of their self-assembly in vivo. One question, not addressed so far, is whether intracellular crystals of proteins can assemble with biopolymers into functional mesocrystal-like structures. During our electron microscopy studies into Artemia franciscana (Crustacea: Branchiopoda), we found initial evidence of such proteinaceous mesostructures. EM preparations with high-pressure freezing and accelerated freeze substitution revealed an extraordinary intracellular source of mesostructured inclusions in both the cyto-and nucleoplasm of the epidermal lining of ovisacs of A. franciscana. Confocal reflection microscopy not only confirmed our finding; it also revealed reflective, light dispersing activity of these flake-like structures, their positioning and orientation with respect to the ovisac inside. Both the striation of alternating electron dense and electron-lucent components and the sharp edges of the flakes indicate self-assembly of material of yet unknown origin under supposed participation of crystallization. However, selected area electron diffraction could not verify the status of crystallization. Energy dispersive X-ray analysis measured a marked increase in nitrogen within the flake-like inclusion, and the almost complete absence of elements that are typically involved in inorganic crystallization. This rise in nitrogen could possibility be related to higher package density of proteins, achieved by mesostructure assembly. The ovisac lining of A. franciscana is endowed with numerous mesostructured inclusions that have not been previously reported. We hypothesize that their self-assembly was from proteinaceous polycrystalline units and carbohydrates. These mesostructured flakes displayed active optical properties, as an umbrella-like, reflective cover of the ovisac, which suggests a functional role in the reproduction of A. franciscana. In turn, studies into ovisac mesostructured inclusions could help to optimizing rearing Artemia as feed for fish farming. We propose Artemia ovisacs as an in vivo model system for studying mesostructure formation.


[Peripheral facial paralysis, polyneuritis and chronic Chagas' disease].
The author reports the case of a 42-year-old woman who developed right peripheral facial paralysis and mixed polyneuritis in the course of an undiagnosed Chagas' disease. The clinical diagnosis could be confirmed by serological reactions for Chagas' disease. A likely relationship between chronic Chagas' disease, peripheral facial paralysis and mixed polyneuritis is discussed.


Peroxiredoxin 1, a Novel HBx-Interacting Protein, Interacts with Exosome Component 5 and Negatively Regulates Hepatitis B Virus (HBV) Propagation through Degradation of HBV RNA.
Hepatitis B virus (HBV) infection is a major risk factor for the development of chronic liver diseases, including cirrhosis and hepatocellular carcinoma (HCC). A growing body of evidence suggests that HBV X protein (HBx) plays a crucial role in viral replication and HCC development. Here, we identified peroxiredoxin 1 (Prdx1), a cellular hydrogen peroxide scavenger, as a novel HBx-interacting protein. Coimmunoprecipitation analysis coupled with site-directed mutagenesis revealed that the region from amino acids 17 to 20 of the HBx, particularly HBx Cys17, is responsible for the interaction with Prdx1. Knockdown of Prdx1 by siRNA significantly increased the levels of intracellular HBV RNA, HBV antigens, and extracellular HBV DNA, whereas knockdown of Prdx1 did not increase the activities of HBV core, enhancer I (Enh1)/X, preS1, and preS2/S promoters. Kinetic analysis of HBV RNA showed that knockdown of Prdx1 inhibited HBV RNA decay, suggesting that Prdx1 reduces HBV RNA levels posttranscriptionally. The RNA coimmunoprecipitation assay revealed that Prdx1 interacted with HBV RNA. The exosome component 5 (Exosc5), a member of the RNA exosome complexes, was coimmunoprecipitated with Prdx1, suggesting its role in regulation of HBV RNA stability. Taken together, these results suggest that Prdx1 and Exosc5 play crucial roles in host defense mechanisms against HBV infection.IMPORTANCE Hepatitis B virus (HBV) infection is a major global health problem. HBx plays important roles in HBV replication and viral carcinogenesis through its interaction with host factors. In this study, we identified Prdx1 as a novel HBx-binding protein. We provide evidence suggesting that Prdx1 promotes HBV RNA decay through interaction with HBV RNA and Exosc5, leading to downregulation of HBV RNA. These results suggest that Prdx1 negatively regulates HBV propagation. Our findings may shed new light on the roles of Prdx1 and Exosc5 in host defense mechanisms in HBV infection.


Radio-chemotherapy of glioblastoma cells promotes phagocytosis by macrophages in vitro.
Immunotherapy is actively explored in glioblastoma (GBM) to improve patient prognosis. Tumor-associated macrophages (TAMs) are abundant in GBM and harnessing their function for anti-tumor immunity is of interest. They are plastic cells that are influenced by the tumor microenvironment, by radio-chemotherapy and by their own phagocytic activity. Indeed, the engulfment of necrotic cells promotes pro-inflammatory (and anti-tumoral) functions while the engulfment of apoptotic cells promotes anti-inflammatory (and pro-tumoral) functions through efferocytosis. To model the effect of radio-chemotherapy on the GBM microenvironment, we exposed human macrophages to supernatant of treated GBM cells in vitro. Macrophages were derived from human monocytes and GBM cells from patient-resected tumors. GBM cells were exposed to therapeutically relevant doses of irradiation and chemotherapy. Apoptosis and phagocytic activity were assessed by flow cytometry. The phagocytic activity of macrophages was increased, and it was correlated with the proportion of apoptotic GBM cells producing the supernatant. Whether uptake of apoptotic tumor cells could occur would depend upon the expression of efferocytosis-associated receptors. Indeed, we showed that efferocytosis-associated receptors, such as AXL, were upregulated. and Perspectives We showed that macrophage phagocytic activity increased when exposed to supernatant from GBM cells treated by radio-chemotherapy. However, as efferocytosis-associated receptors were up-regulated, this effect could be deleterious for the anti-GBM immune response. We speculate that by inducing GBM cell apoptosis in parallel to an increase in efferocytosis receptor expression, the impact of radio-chemotherapy on phagocytic activity could promote anti-inflammatory and pro-tumoral TAM functions.


Physical Preparation Strategies of Professional Jockeys.
Kiely, M, Warrington, G, McGoldrick, A, and Cullen, S. Physical preparation strategies of professional jockeys. J Strength Cond Res 36(11): 3184-3189, 2022-Professional horse racing is a physically demanding sport. The aim of the study was to examine the physical preparation strategies of jockeys for racing. A questionnaire was developed and validated which comprised of 4 sections; (a) background information, (b) making weight and current associated exercise habits, (c) current physical activity practices, and (d) jockey perceptions of strength and conditioning (S&C) and current practices. Eighty-five jockeys ( n = 38 professional flat, n = 47 professional jump) completed the questionnaire in race course weighing rooms representing 80% of the professional athlete population. In total, 77.6% of jockeys participate in physical activity outside of riding. Jockeys that participated in S&C (42.4%) reported their most frequent type of S&C practice; cardio (52.8%), high-intensity interval training (33.3%), flexibility and mobility training (8.3%), resistance training (5.6%). There was no significant difference in S&C participation between total flat and total jump licenses ( p = 0.530; [PHI] = 0.068). Difficulty making weight was reported by 55.3% of jockeys. Exercise alone was used by 29.4% of jockeys to rapidly reduce weight. There was no significant difference ( p = 0.201, [PHI] = 0.357) between the frequency of rapid weight loss per month for total flat (1.7 ± 1.7) and total jump jockeys (1.6 ± 0.5). This study represents the only published data on the physical preparation strategies of jockeys. Jockeys do not partake in physical activity, which mimics the repeated high-intensity demands of racing. Future research is required to examine the effects of specific S&C interventions on riding performance.


Optical Control of the Resonant Dipole-Dipole Interaction between Rydberg Atoms.
We report on the local control of the transition frequency of a spin 1/2 encoded in two Rydberg levels of an individual atom by applying a state-selective light shift using an addressing beam. With this tool, we first study the spectrum of an elementary system of two spins, tuning it from a nonresonant to a resonant regime, where "bright" (super-radiant) and "dark" (subradiant) states emerge. We observe the collective enhancement of the microwave coupling to the bright state. We then show that after preparing an initial single spin excitation and letting it hop due to the spin-exchange interaction, we can freeze the dynamics at will with the addressing laser, while preserving the coherence of the system. In the context of quantum simulation, this scheme opens exciting prospects for engineering inhomogeneous XY spin Hamiltonians or preparing spin-imbalanced initial states.


18F-FDG PET response and clinical outcomes after stereotactic body radiation therapy for metastatic melanoma.
Clinical data that support stereotactic body radiation therapy (SBRT) metastatic malignant melanoma (MM) are limited. Furthermore, functional imaging with 18F-fludeoxyglucose positron emission tomography (PET) may offer a more accurate post-SBRT assessment. Therefore, we assessed the clinical outcomes and metabolic response of metastatic MM after SBRT. Patients with MM who were treated with SBRT and had pre- and post-PET scans (>1) were included in this study. A total of 390 pre- and post-SBRT PET/computed tomography (CT) scans for 80 metastases were analyzed. The PET metabolic response was evaluated per the PET Response Criteria in Solid Tumors (PERCIST), version 1.0, criteria. Single-fraction equivalent dose (SFED) was calculated as per the standard. The Kaplan-Meier method was used for estimates of overall survival (OS) and progression-free survival. The cumulative incidence method was used to estimate metastasis control (MC). A Wilcoxon test was used to compare survival estimates. The prognostic factors for MC and OS were assessed using the Cox proportional hazards model, and the Likelihood Ratio was also used for comparisons between groups. A median of 6 PET scans (range, 2-6 scans) was evaluated for each metastasis. The median SFED was 42.8 Gy (range, 18-56.4 Gy) and the median biologically effective dose was 254.4 Gy2.5 (range, 100.8-540 Gy2.5). Twenty percent of patients received chemotherapy and 59% received immunotherapy: granulocyte-macrophage colony-stimulating factor (64%) and ipilimumab (34%). MC was 94% and 90% at 1 year and 3 years, respectively. The OS was 74% and 27% and 1 year and 3 years, respectively. Complete response was achieved in 90% at a median of 2.8 months (range, 0.4-25.2 months). SFED >24 Gy correlated with improved MC (93% vs 75%, P = .01). Acute and late grade 3+ toxicities were 4% and 11%, respectively, with no grade 5 toxicity. Post-SBRT PET/CT for extracranial metastatic MM resulted in high rates of complete response at a median of 2.8 months, and durable MC was achieved with SFED >24 Gy. SBRT, in addition to surgery and ablation, should be discussed with patients with MM, especially those with oligometastases.


Same-sex sexual behavior and evolution.
Same-sex sexual behavior has been extensively documented in non-human animals. Here we review the contexts in which it has been studied, focusing on case studies that have tested both adaptive and non-adaptive explanations for the persistence of same-sex sexual behavior. Researchers have begun to make headway unraveling possible evolutionary origins of these behaviors and reasons for their maintenance in populations, and we advocate expanding these approaches to examine their role as agents of evolutionary change. Future research employing theoretical, comparative and experimental approaches could provide a greater understanding not only of how selection might have driven the evolution of same-sex sexual behaviors but also ways in which such behaviors act as selective forces that shape social, morphological and behavioral evolution.


CO Gas Induced Phase Separation in PtPb@Pt Catalyst and Formation of Ultrathin Pb Nanosheets Probed by In Situ Transmission Electron Microscopy.
PtPb@Pt catalysts are very useful and widely applied in various industrial reactions. Here, the phase stability of such catalysts is compared in both CO gas and vacuum conditions at elevated temperatures using aberration-corrected in situ transmission electron microscopy (TEM). A Pt aggregation process takes place affected by CO gas, which results in direct exposure of the PtPb core to CO. A phase separation process, in which Pb atoms are stripped off the original PtPb@Pt nanoparticles, is unambiguously identified in CO gas. At initial stages, the as nucleated Pb islands are amorphous. Once the ultrathin Pb islands reach ≈3.5 nm or higher, they suddenly became crystalline. The interaction between Pb and CO gas stabilizes the ultrathin Pb nanosheets, resulting in the formation of a large quantity of Pb nanosheets and Pb-depleted PtPb0.08 nanoparticles. In sharp contrast, when heated up in a vacuum, the PtPb@Pt catalyst remains intact. The results of this study shine light on the "toxic" effect of CO that results in failures of many Pt-based catalysts and discloses formation mechanism of ultrathin Pb nanosheets.


Blood coagulation changes in mice bearing Lewis lung carcinoma, a metastasizing tumor.
In view of the possible role of platelets and coagulation mechanisms in the growth and dissemination of solid tumors, a number of hematological parameters were followed during development of an experimental syngeneic tumor in mice, Lewis lung carcinoma. This tumor, when transplanted i.m. in C57BL/6 mice, grows locally and spontaneously metastasizes to the lungs. The transplanted animals survive for about 4 weeks. Metastases are visible from the third week. A slight but constant increase in plasma fibrinogen level and marked thrombocytopenia were first observed during the second week after tumor implantation. No other significant changes in coagulation and fibrinolysis parameters were detected. Moreover, the animals developed marked hemolytic anemia, possibly microangiopathic in origin. 125I-labelled fibrinogen survival was decreased by about 20% during the second week after tumor implantation and was not further reduced later. Fibrinogen turnover was progressively accelerated, being more than doubled by the end of the third week. Labeled fibrinogen accumulated in the primary tumor and in the lungs (its rate of disappearance from the tumor was much slower than that from lungs or blood). 51Cr-labeled platelet survival did not change throughout the observation period, whereas platelet turnover was markedly reduced from the end of the second week, suggesting defective platelet production. 51Cr-labeled RBC survival was drastically reduced to about 30% of the controls starting from the second week. The occurence of low-grade, localized intravascular coagulation could be suggested on the basis of these data. Moreover, when Lewis lung carcinoma cells were abruptly injected i.v. through the tall vein, more impressive signs of intravascular coagulation could be seen. Indeed, there was a rapid decrease in the number of platelets, a reduction in fibrinogen, and an increase in fibrin-fibrinogen degradation products. The effects of i.v. injection of Lewis lung carcinoma cells indicate a relevant interference of cancer cells with the hematostatic system. In contrast, the tenuous evidence fo coagulation disorders in animals receiving injections of tumor cells i.m. seems to indicate a limited effect on hemostasis of the same cells during i.m. tumor growth.


Salivary cortisol levels and allergy in children: the ALADDIN birth cohort.
Pre- and postnatal stress have been related to allergy in children, but evidence from prospective studies is limited. Several environmental factors can influence the salivary cortisol level, which is used as a measure of activity of the hypothalamic-pituitary-adrenal axis. The aim of this study was to assess the association between salivary cortisol levels at 6 months of age and allergic manifestations during the first 2 years of life. Salivary samples for the analysis of cortisol level were collected at 6 months of age on 3 occasions during 1 day from 203 children. Blood samples were collected at 6, 12, and 24 months of age for analyses of specific IgE. Information on allergy-related symptoms was obtained by repeated examinations of the children. Generalized estimating equation statistics were used to calculate the overall risk for outcome measures. The adjusted odds ratio for the relationship between morning cortisol level and IgE sensitization was 1.60 (95% CI, 1.22-2.10, P = .001) and for eczema it was 1.28 (95% CI, 1.03-1.59, P = .026). The odds ratio for afternoon cortisol level in relation to sensitization and eczema was 1.56 (95% CI, 1.26-1.94, P < .001) and 1.33 (95% CI, 1.12-1.58, P = .001), respectively, and for evening cortisol level it was 1.49 (95% CI, 1.22-1.83, P < .001) and 1.37 (95% CI, 1.18-1.59, P < .001). Salivary cortisol level in the evening was associated with food allergy. The association between salivary cortisol levels in infancy and allergic sensitization and allergic symptoms suggests a role of an altered hypothalamic-pituitary-adrenal axis in the etiological process of allergies.


Identification and characterization of a novel enhancer for the human MCT-1 oncogene promoter.
Cloning and characterization of the promoter region for the MCT-1 oncogene is described. We used luciferase assays to identify cis-acting elements responsible for human MCT-1 promoter function. The MCT-1 promoter is TATA-less with a consensus initiator element located at the transcription start site and facilitated by two Sp1 sites that directs basal transcription. Deletion of a region of the MCT-1 promoter (-133 to -122) resulted in significant decrease in luciferase activity, suggesting that this region contains a positive cis-acting element. Using mobility shift assays with a 26-mer oligonucleotide, which contains this fragment and its flanking regions, we demonstrated the presence of sequence-specific DNA-binding protein in both Jurkat and Hela nuclear extracts that we designated as LMBF (for lymphoid MCT-1 binding factor). This 26-mer oligonucleotide containing the LMBF binding site is required for maximum transcriptional activity of the MCT-1 promoter. Although the 26-mer oligonucleotide contains a sequence with strong homology to a heat-shock factor consensus, competitive electrophoretic mobility shift assay (EMSA) analysis demonstrated that the binding protein is not a known member of heat shock family. Furthermore, this sequence when placed in reverse orientation downstream of the luciferase gene was able to enhance luciferase activity driven by a minimal promoter. These data are consistent with this sequence behaving as an enhancer. Finally, Southwestern blot analysis revealed a 96-kDa protein capable of binding a probe containing the LMBF binding site.


Muscle contraction and the elasticity-mediated crosstalk effect.
Cooperative action of molecular motors is essential for many cellular processes. One possible regulator of motor coordination is the elasticity-mediated crosstalk (EMC) coupling between myosin II motors whose origin is the tensile stress that they collectively generate in actin filaments. Here, we use a statistical mechanical analysis to investigate the influence of the EMC effect on the sarcomere -- the basic contractile unit of skeletal muscles. We demonstrate that the EMC effect leads to an increase in the attachment probability of motors located near the end of the sarcomere while simultaneously decreasing the attachment probability of the motors in the central part. Such a polarized attachment probability would impair the motors' ability to cooperate efficiently. Interestingly, this undesired phenomenon becomes significant only when the system size exceeds that of the sarcomere in skeletal muscles, which provides an explanation for the remarkable lack of sarcomere variability in vertebrates. Another phenomenon that we investigate is the recently observed increase in the duty ratio of the motors with the tension in muscle. We reveal that the celebrated Hill's equation for muscle contraction is very closely related to this observation.


Keeping it between us: Managerial endorsement of public versus private voice.
When employees use public settings such as team meetings to engage in voice-the expression of work ideas or concerns, they can spur useful discussions, action planning, and problem solving. However, we make the case that managers, whose support is essential for voice to have a functional impact, are averse to publicly expressed voice and prefer acting on voice that is privately brought up to them in one-on-one settings. Drawing on face management theory (Goffman, 1967), we argue that voice expressed in front of an audience, compared with that expressed one-on-one, is more threatening to the image that managers seek to portray as competent and unerring leaders, and that leads managers to respond more defensively to public voice and endorse it less. This, we propose, is especially true when the relationship quality between manager and employee is weak as public voice from relationally distant employees is perceived as a stronger challenge. Across five studies (correlational and experimental), we find support for our arguments and rule out alternative explanations such as that managers are aversive to public voice because it threatens their ego or that managers feel more accountable to act on publicly provided input. We discuss the implications of our findings for theory and practice. (PsycInfo Database Record (c) 2021 APA, all rights reserved).


Dipeptidylpeptidase--IV, a key enzyme for the degradation of incretins and neuropeptides: activity and expression in the liver of lean and obese rats.
Given the scarcity of donors, moderately fatty livers (FLs) are currently being considered as possible grafts for orthotopic liver transplantation (OLT), notwithstanding their poor tolerance to conventional cold preservation. The behaviour of parenchymal and sinusoidal liver cells during transplantation is being studied worldwide. Much less attention has been paid to the biliary tree, although this is considered the Achille's heel even of normal liver transplantation. To evaluate the response of the biliary compartment of FLs to the various phases of OLT reliable markers are necessary. Previously we demonstrated that Alkaline Phosphatase was scarcely active in bile canaliculi of FLs and thus ruled it out as a marker. As an alternative, dipeptidylpeptidase-IV (DPP-IV), was investigated. This ecto-peptidase plays an important role in glucose metabolism, rapidly inactivating insulin secreting hormones (incretins) that are important regulators of glucose metabolism. DPP-IV inhibitors are indeed used to treat Type II diabetes. Neuropeptides regulating bile transport and composition are further important substrates of DPP-IV in the enterohepatic axis. DPP-IV activity was investigated with an azo-coupling method in the liver of fatty Zucker rats (fa/fa), using as controls lean Zucker (fa/+) and normal Wistar rats. Protein expression was studied by immunofluorescence with the monoclonal antibody (clone 5E8). In Wistar rat liver, DPP-IV activity and expression were high in the whole biliary tree, and moderate in sinusoid endothelial cells, in agreement with the literature. Main substrates of DPP-IV in hepatocytes and cholangiocytes could be incretins GLP-1 and GIP, and neuropeptides such as vasoactive intestinal peptide (VIP) and substance P, suggesting that these substances are inactivated or modified through the biliary route. In lean Zucker rat liver the enzyme reaction and protein expression patterns were similar to those of Wistar rat. In obese rat liver the patterns of DPP-IV activity and expression in hepatocytes reflected the morphological alterations induced by steatosis as lipid-rich hepatocytes had scarce activity, located either in deformed bile canaliculi or in the sinusoidal and lateral domains of the plasma membrane. These findings suggest that bile canaliculi in steatotic cells have an impaired capacity to inactivate incretins and neuropeptides. Incretin and/or neuropeptide deregulation is indeed thought to play important roles in obesity and insulin-resistance. No alteration in enzyme activity and expression was found in the upper segments of the biliary tree of obese respect to lean Zucker and Wistar rats. In conclusion, this research demonstrates that DPP-IV is a promising in situ marker of biliary functionality not only of normal but also of fatty rats. The approach, initially devised to investigate the behaviour of the liver during the various phases of transplantation, appears to have a much higher potentiality as it could be further exploited to investigate any pathological or stressful conditions involving the biliary tract (i.e., metabolic syndrome and cholestasis) and the response of the biliary tract to therapy and/or to surgery.


Effects of GSM modulated radio-frequency electromagnetic radiation on permeability of blood-brain barrier in male & female rats.
With the increased use of mobile phones, their biological and health effects have become more important. Usage of mobile phones near the head increases the possibility of effects on brain tissue. This study was designed to investigate the possible effects of pulse modulated 900MHz and 1800MHz radio-frequency radiation on the permeability of blood-brain barrier of rats. Study was performed with 6 groups of young adult male and female wistar albino rats. The permeability of blood-brain barrier to intravenously injected evans blue dye was quantitatively examined for both control and radio-frequency radiarion exposed groups. For male groups; Evans blue content in the whole brain was found to be 0.08±0.01mg% in the control, 0.13±0.03mg% in 900MHz exposed and 0.26±0.05mg% in 1800MHz exposed animals. In both male radio-frequency radiation exposed groups, the permeability of blood-brain barrier found to be increased with respect to the controls (p<0.01). 1800MHz pulse modulated radio-frequency radiation exposure was found more effective on the male animals (p<0.01). For female groups; dye contents in the whole brains were 0.14±0.01mg% in the control, 0.24±0.03mg% in 900MHz exposed and 0.14±0.02mg% in 1800MHz exposed animals. No statistical variance found between the control and 1800MHz exposed animals (p>0.01). However 900MHz pulse modulated radio-frequency exposure was found effective on the permeability of blood-brain barrier of female animals. Results have shown that 20min pulse modulated radio-frequency radiation exposure of 900MHz and 1800MHz induces an effect and increases the permeability of blood-brain barrier of male rats. For females, 900MHz was found effective and it could be concluded that this result may due to the physiological differences between female and male animals. The results of this study suggest that mobile phone radation could lead to increase the permeability of blood-brain barrier under non-thermal exposure levels. More studies are needed to demonstrate the mechanisms of that breakdown.


Purification and properties of adenosine triphosphate-creatine phosphotransferase from muscle of the dogfish Scylliorhinus canicula.
1. Creatine kinase occurs in high concentration in the soluble proteins of dogfish muscle. A fourfold purification gives essentially pure enzyme but with a low specific activity. This appears to be a property of the native enzyme and not a result of the isolation procedures used. 2. The amino acid composition is similar to that of other phosphagen kinases, but the enzyme differs from mammalian creatine kinases in having four thiol groups readily reactive towards 5,5'-dithiobis-(2-nitrobenzoic acid). Titration of two thiol groups is accompanied by almost complete loss of activity. The remaining two thiol groups react at different rates, suggesting that modifying the third thiol group affects the reactivity of the fourth thiol group. 3. The enzyme is markedly protected against inactivation by iodoacetamide by MgATP or MgADP. Addition of creatine to MgADP decreases protection, but the further addition of Cl(-) restores protection to the original value. The quaternary MgADP-creatine-enzyme-nitrate complex protects very strongly as is found for the rabbit enzyme. The involvement of the conformational state of the enzyme in such effects is discussed. 4. Creatine kinase from both dogfish and rabbit is equally sensitive to urea denaturation. Urea protects the dogfish enzyme by about 9% against inhibition by iodoacetamide. 5. The formation of a hybrid between the dogfish and rabbit enzymes in vitro has been demonstrated. 6. At high substrate concentrations the dogfish enzyme shows apparent ordered kinetics. The effect of temperature on V(max.) and the Michaelis constants for MgATP and creatine were determined. These and changes in the apparent activation energy suggest that limited adaptation has occurred commensurate with physiological need.


Pulsed Electromagnetic Field Versus Whole Body Vibration on Cartilage and Subchondral Trabecular Bone in Mice With Knee Osteoarthritis.
Pulsed electromagnetic field (PEMF) and whole body vibration (WBV) interventions are expected to be important strategies for management of osteoarthritis (OA). The aim of the study was to investigate the comparative effectiveness of PEMF versus WBV on cartilage and subchondral trabecular bone in mice with knee OA (KOA) induced by surgical destabilization of the medial meniscus (DMM). Forty 12-week-old male C57/BL mice were randomly divided into four groups (n = 10): Control, OA, PEMF, and WBV. OA was induced (OA, PEMF, and WBV groups) by surgical DMM of right knee joint. Mice in PEMF group received 1 h/day PEMF exposure with 75 Hz, 1.6 mT for 4 weeks, and the WBV group was exposed to WBV for 20 min/day with 5 Hz, 4 mm, 0.3 g peak acceleration for 4 weeks. Micro-computed tomography (micro-CT), histology, and immunohistochemistry analyses were performed to evaluate the changes in cartilage and microstructure of trabecular bone. The bone volume fraction (BV/TV), trabecular thickness (Tb.Th), and trabecular number (Tb.N) increased, and bone surface/bone volume (BS/BV) decreased by micro-CT analysis in PEMF and WBV groups. The Osteoarthritis Research Society International (OARSI) scores in PEMF and WBV groups were significantly lower than in the OA group. Immunohistochemical results showed that PEMF and WBV promoted expressions of Aggrecan, and inhibited expressions of IL-1β, ADAMTS4, and MMP13. Superior results are seen in PEMF group compared with WBV group. Both PEMF and WBV were effective, could delay cartilage degeneration and preserve subchondral trabecular bone microarchitecture, and PEMF was found to be superior to WBV. Bioelectromagnetics. 2020;41:298-307 © 2020 Bioelectromagnetics Society.


Biological and clinical aspects in laser hair removal.
In the past century, unwanted hair has been traditionally treated with multitudes of techniques that were found to be slow, tedious, painful, impractical, and resulted in poor long-term efficacy. Consequently, there has been a public demand for a novel, rapid, reliable, safe, and affordable hair removal technique. In the last decade, laser and light-based technology for hair removal became one of the fastest growing procedures in modern cosmetic dermatology. To discuss the latest scientific and clinical issues in the field of photoepilation as evolved in the past decade: hair biology, laser physics and skin optics, technology and clinical experience. From substantial clinical experience, it becomes apparent that in the ideal subject with fair skin and dark hair, a single treatment can reduce hair by 10-40%; three treatments by 30-70%; and repeated treatments by as much as 90%. These results persist for as long as 12 months. Diffuse and perifollicular cutaneous erythema and pigmentary changes are the most common adverse side effects. Most complications are generally temporary. Photoepilation, when properly used, offers clear advantages when compared with older, traditional techniques. Although an ever-increasing number of published studies have confirmed the safety and short and long-term efficacy of photoepilation, the technology still has limits and risks.


[Influence of berotec inhalations on pulmonary gas exchange in patients with bronchial asthma].
To determine mechanisms inducing changes in lung gas exchange after berotec inhalation in patients with bronchial asthma (BA). A study was made of external respiration function including estimation of lung transfer factor (TLCO sbh.) and its components--transfer coefficient (KCO), membrane diffusion capacity (DM), pulmonary capillary blood volume (Vc)--by the single breath holding method on CO in 14 healthy persons and 39 patients with BA before and after berotec inhalation. The data were obtained in the absolute values and their changes after berotec inhalation were measured in percent to initial ones. In healthy subjects we did not observe changes of TLCO sbh. and its components after berotec inhalation, only an insignificant decrease of VA eff. was registered (p < 0.01). After berotec inhalation a considerable improvement in Sgaw, FEV1, RPV and a decrease of TLCO sbh., KCO were observed. Changes in TLCO sbh. were caused by a decrease in DM without changes in VA eff. in BA patients with initial normal values of KCO. In patients with smaller initial values of KCO moderate improvement of bronchial permeability without changes in KCO, PaO2 were observed. These changes occurred due to a decrease of VA eff. Changes of the transfer factor and its components in patients with BA reflect two types of adaptation reactions in response to berotec inhalation and their role in mechanisms of compensation of gas exchange in the lungs.


Multivariate modeling of club drug use initiation among gay and bisexual men.
This paper documents patterns and sequence of initiation of club drug use in a sample of 450 gay and bisexual men in New York City. Quantitative and qualitative baseline data from a year-long longitudinal investigation conducted between 2001 and 2005 were analyzed. The study focused on the use of five club drugs-cocaine, GHB, ketamine, ecstasy, and methamphetamine-using self-reported indications of use for a period of 4 months prior to assessment. Patterns of club drug use among gay and bisexual demonstrated that poly-club-drug use is common, and that patterns of use can be differentiated along the lines of age, race/ethnicity, and sexual orientation, with those who are older, Black, and bisexual, reporting less club drug use. The majority of the men initiated use of the five club drugs as follows: (a) cocaine, (b) ecstasy, (c) ketamine, (d) methamphetamine, and (e) GHB. Variations in patterns were related to both age and level of poly-club-drug use. The sequencing and/or patterns of club drug use may be better explained by socialization processes in the gay community than by Gateway Theory, which has been traditionally used to explain patterns of drug use in the population. Future research should more closely examine the synergy of drug use combinations with an emphasis on measuring the extent to which the drugs are taken in synchronicity.


This new field of inclusive education: beginning a dialogue on conceptual foundations.
Numerous scholars have suggested that the standard knowledge base of the field of special education is not a suitable intellectual foundation for the development of research, policy, and practice in the field of inclusive education. Still, we have yet to have a dialogue on what conceptual foundations may be most generative for the growth and development of the field of inclusive education. This article imagines and initiates such a new dialogue among educational researchers and teacher educators about the intellectual resources that can best support inclusive educators everywhere. As inclusive education gets increasingly taken up within international policy discourses, it may be imperative to explore and identify theories and ideas that can be responsive to diverse and hugely unequal contexts of schooling. This article forwards an initial collection of intellectual resources for an inclusive education that can accommodate such complex schooling conditions and invites rich scholarly exchange on this issue.


Human vascular endothelial cells stimulate a lower frequency of alloreactive CD8+ pre-CTL and induce less clonal expansion than matching B lymphoblastoid cells: development of a novel limiting dilution analysis method based on CFSE labeling of lymphocytes.
We have previously shown that human endothelial cells (EC) are less efficient than professional APC, e.g., B lymphoblastoid cells (BLC), at stimulating allogeneic CD8(+) T cells to develop into CTL. In this study we describe FACS-based limiting dilution analyses using the dilution of the intracellular dye CFSE as an indicator of CD8(+) T cell alloactivation and expansion with significantly increased sensitivity compared with conventional, cytotoxicity-based assays. In addition, this assay permits the relative size of clonal CTL populations that are generated in individual CD8(+) T cell cultures to be determined (clonal burst size). We have applied this method to quantitatively compare the generation of CTL at the clonal level following stimulation of allogeneic CD8(+) T cells by either BLC or HUVEC derived from the same donor. CD8(+) T cells expanded by allostimulation were identified as CD8(+), CFSE(low) cells and were categorized as CTL by the expression of intracellular perforin and IFN-gamma. Precursor frequencies for EC-stimulated CTL were 5- to 40-fold (mean, 7.5-fold) lower compared with BLC-stimulated CTL (p < 0.01). Concomitantly, the average clonal burst sizes in EC-stimulated CTL cultures were significantly smaller than those in conventional CTL cultures, primarily due to the occurrence of some very large clone sizes exclusively with BLC stimulation. Although EC-stimulated CTL were generated only from the memory subset of CD8(+) T cells, BLC-stimulated very large burst sizes of CTL were observed from both naive and memory CD8(+) T cell precursors. These data establish that both a lower frequency of reactive precursors and more limited clonal expansion, but not regulatory T cells, contribute to the reduced capacity of EC to promote alloreactive CTL differentiation compared with that of professional APC.


Exploring the benefits of bilateral cochlear implants.
Several recent reports indicate that both localization and speech intelligibility in spatially separated noise are substantially improved by using cochlear implants (CIs) in both ears rather than in just one. Benefits appear to be largely derived from the effects of level variations at the two ears due to the head shadow whereas contributions from interaural time differences (ITDs) seem smaller than in normal hearing listeners. The effect of binaural unmasking estimated from speech studies to date varies from study to study and is possibly confounded by issues such as listening experience, bias or loudness effects when comparing the performance for the better ear with that using both ears. To improve the contribution from timing information at the two ears, it may be necessary to change present clinical sound-processing schemes that currently preserve only envelope cues so that they also preserve fine-timing information. However, recently published data show that basic psychophysical sensitivity to fine-timing ITDs in CI patients is very poor for rates beyond a few hundred hertz, suggesting that subjects do not actually hear ITD cues at those rates anyway. Data from a number of new studies are presented to discuss these and other issues related to the potential to benefit from bilateral implantation.


Bone marrow niches in haematological malignancies.
Haematological malignancies were previously thought to be driven solely by genetic or epigenetic lesions within haematopoietic cells. However, the niches that maintain and regulate daily production of blood and immune cells are now increasingly being recognized as having an important role in the pathogenesis and chemoresistance of haematological malignancies. Within haematopoietic cells, the accumulation of a small number of recurrent mutations initiates malignancy. Concomitantly, specific alterations of the niches, which support haematopoietic stem cells and their progeny, can act as predisposition events, facilitating mutant haematopoietic cell survival and expansion as well as contributing to malignancy progression and providing protection of malignant cells from chemotherapy, ultimately leading to relapse. In this Perspective, we summarize our current understanding of the composition and function of the specialized haematopoietic niches of the bone marrow during health and disease. We discuss disease mechanisms (rather than malignancy subtypes) to provide a comprehensive description of key niche-associated pathways that are shared across multiple haematological malignancies. These mechanisms include primary driver mutations in bone marrow niche cells, changes associated with increased hypoxia, angiogenesis and inflammation as well as metabolic reprogramming by stromal niche cells. Consequently, remodelling of bone marrow niches can facilitate immune evasion and activation of survival pathways favouring malignant haematopoietic cell maintenance, defence against excessive reactive oxygen species and protection from chemotherapy. Lastly, we suggest guidelines for the handling and biobanking of patient samples and analysis of the niche to ensure that basic research identifying therapeutic targets can be more efficiently translated to the clinic. The hope is that integrating knowledge of how bone marrow niches contribute to haematological disease predisposition, initiation, progression and response to therapy into future clinical practice will likely improve the treatment of these disorders.


[Stimulating effect of gamma-aminobutyric acid on rat hippocampal neurons in vitro].
Effects of GABA on the background and electrically stimulated activity of single neurons and population spikes were investigated in isolated hippocampal slices. Application of relatively large GABA concentrations (10(-3) mol/l and more) depressed an antidromic population spike, field EPSP and neuronal background activity. Low concentrations of GABA (less than 10(-3) mol/l) added to the bath increased the population spikes amplitude and the late component of field EPSP, facilitated single neurone responses, their background activity and epileptiform discharges. GABA-evoked depolarization was observed in the majority of the studied neurons. The duality of the GABA action on central neurons are discussed.


Clonal Expansion and Interrelatedness of Distinct B-Lineage Compartments in Multiple Myeloma Bone Marrow.
Multiple myeloma is characterized by the clonal expansion of malignant plasma cells in the bone marrow. But the phenotypic diversity and the contribution of less predominant B-lineage clones to the biology of this disease have been controversial. Here, we asked whether cells bearing the dominant multiple myeloma immunoglobulin rearrangement occupy phenotypic compartments other than that of plasma cells. To accomplish this, we combined 13-parameter FACS index sorting and t-Stochastic Neighbor Embedding (t-SNE) visualization with high-throughput single-cell immunoglobulin sequencing to track selected B-lineage clones across different stages of human B-cell development. As expected, the predominant clones preferentially mapped to aberrant plasma cell compartments, albeit phenotypically altered from wild type. Interestingly, up to 1.2% of cells of the predominant clones colocalized with B-lineage cells of a normal phenotype. In addition, minor clones with distinct immunoglobulin sequences were detected in up to 9% of sequenced cells, but only 2 out of 12 of these clones showed aberrant immune phenotypes. The majority of these minor clones showed intraclonal silent nucleotide differences within the CDR3s and varying frequencies of somatic mutations in the immunoglobulin genes. Therefore, the phenotypic range of multiple myeloma cells in the bone marrow is not confined to aberrant-phenotype plasma cells but extends to low frequencies of normal-phenotype B cells, in line with the recently reported success of B cell-targeting cellular therapies in some patients. The majority of minor clones result from parallel nonmalignant expansion. Cancer Immunol Res; 5(9); 744-54. ©2017 AACR.


NKT cells act through third party bone marrow-derived cells to suppress NK cell activity in the liver and exacerbate hepatic melanoma metastases.
Uveal melanoma (UM) is the most common intraocular tumor in adults and liver metastasis is the leading cause of death in UM patients. We have previously shown that NKT cell-deficient mice develop significantly fewer liver metastases from intraocular melanomas than do wild-type (WT) mice. Here, we examine the interplay between liver NKT cells and NK cells in resistance to liver metastases from intraocular melanomas. NKT cell-deficient CD1d(-/-) mice and WT C57BL/6 mice treated with anti-CD1d antibody developed significantly fewer liver metastases than WT mice following either intraocular or intrasplenic injection of B16LS9 melanoma cells. The increased number of metastases in WT mice was associated with reduced liver NK cytotoxicity and decreased production of IFN-γ. However, liver NK cell-mediated cytotoxic activity was identical in non-tumor bearing NKT cell-deficient mice and WT mice, indicating that liver metastases were crucial for the suppression of liver NK cells. Depressed liver NK cytotoxicity in WT mice was associated with production of IL-10 by bone marrow-derived liver cells that were neither Kupffer cells nor myeloid-derived suppressor cells and by increased IL-10 receptor expression on liver NK cells. IL-10(-/-) mice had significantly fewer liver metastases than WT mice, but were not significantly different from NKT cell-deficient mice. Thus, development of melanoma liver metastases is associated with upregulation of IL-10 in the liver and an elevated expression of IL-10 receptor on liver NK cells. This impairment of liver NK activity is NKT cell-dependent and only occurs in hosts with melanoma liver metastases.


Differential Expression of Immune Genes between Two Closely Related Beetle Species with Different Immunocompetence following Attack by Asecodes parviclava.
Endoparasitoid wasps are important natural enemies of many insect species and are major selective forces on the host immune system. Despite increased interest in insect antiparasitoid immunity, there is sparse information on the evolutionary dynamics of biological pathways and gene regulation involved in host immune defense outside Drosophila species. We de novo assembled transcriptomes from two beetle species and used time-course differential expression analysis to investigate gene expression differences in closely related species Galerucella pusilla and G. calmariensis that are, respectively, resistant and susceptible against parasitoid infection by Asecodes parviclava parasitoids. Approximately 271 million and 224 million paired-ended reads were assembled and filtered to form 52,563 and 59,781 transcripts for G. pusilla and G. calmariensis, respectively. In the whole-transcriptome level, an enrichment of functional categories related to energy production, biosynthetic process, and metabolic process was exhibited in both species. The main difference between species appears to be immune response and wound healing process mounted by G. pusilla larvae. Using reciprocal BLAST against the Drosophila melanogaster proteome, 120 and 121 immune-related genes were identified in G. pusilla and G. calmariensis, respectively. More immune genes were differentially expressed in G. pusilla than in G. calmariensis, in particular genes involved in signaling, hematopoiesis, and melanization. In contrast, only one gene was differentially expressed in G. calmariensis. Our study characterizes important genes and pathways involved in different immune functions after parasitoid infection and supports the role of signaling and hematopoiesis genes as key players in host immunity in Galerucella against parasitoid wasps.


CD8+CD28-CD127loCD39+ regulatory T-cell expansion: A new possible pathogenic mechanism for HIV infection?
HIV-associated immunodeficiency is related to loss of CD4+ T cells. This mechanism does not explain certain manifestations of HIV disease, such as immunodeficiency events in patients with greater than 500 CD4+ T cells/μL. CD8+CD28-CD127loCD39+ T cells are regulatory T (Treg) lymphocytes that are highly concentrated within the tumor microenvironment and never analyzed in the circulation of HIV-infected patients. We sought to analyze the frequency of CD8+CD28-CD127loCD39+ Treg cells in the circulation of HIV-infected patients. The frequency of circulating CD8+CD28-CD127loCD39+ Treg cells was analyzed and correlated with viral load and CD4+ T-cell counts/percentages in 93 HIV-1-infected patients subdivided as follows: naive (n = 63), elite controllers (n = 19), long-term nonprogressors (n = 7), and HIV-infected patients affected by tumor (n = 4). The same analyses were performed in HIV-negative patients with cancer (n = 53), hepatitis C virus-infected patients (n = 17), and healthy donors (n = 173). HIV-infected patients had increased circulating levels of functional CD8+CD28-CD127loCD39+ Treg cells. These cells showed antigen specificity against HIV proteins. Their frequency after antiretroviral therapy (ART) correlated with HIV viremia, CD4+ T-cell counts, and immune activation markers, suggesting their pathogenic involvement in AIDS- or non-AIDS-related complications. Their increase after initiation of ART heralded a lack of virologic or clinical response, and hence their monitoring is clinically relevant. HIV infection induces remarkable expansion of CD8+CD28-CD127loCD39+ Treg cells, the frequency of which correlates with both clinical disease and signs of chronic immune cell activation. Monitoring their frequency in the circulation is a new marker of response to ART when effects on viremia and clinical response are not met.