High Throughput and Noninvasive Exosomal PD-L1 Detection for Accurate Immunotherapy Response Prediction via Tim4-Functionalized Magnetic Core-Shell Metal-Organic Frameworks.
Exosomal PD-L1 has been increasingly considered a noninvasive and accurate predictive marker for immunotherapy treatment response. However, the clinical monitoring of exosomal PD-L1 expression is still limited by its complex biological environment as well as the lack of a robust isolation strategy. Here, a Tim4-functionalized magnetic core-shell metal-organic framework (denoted as Fe3O4@SiO2-ILI-01@Tim4) was facilely constructed via layer-by-layer assembly. Owing to the strongly hydrophilic organic ligand of 1,3-bis(4-carboxybutyl)imidazolium bromide (ILI), magnetic Fe3O4@SiO2-ILI-01@Tim4 was endowed with the merits of low nonspecific adsorption and quick, easy, and convenient isolation of exosomes. The capture efficiency of Fe3O4@SiO2-ILI-01@Tim4 reached as high as 90.3 ± 0.5% and the recovery rate for exosomes was up to 93.0 ± 6.1%. The purity of the isolated exosomes was 7.5 times higher than that via the ultracentrifugation (UC) method. By further combination with immunofluorescence assay, high throughput and noninvasive exosomal PD-L1 detection for accurate immunotherapy response prediction was achieved. The prognosis accuracy of the developed Fe3O4@SiO2-ILI-01@Tim4-based strategy reached 85.7%, whereas the prognosis accuracy of the clinical gold standard, the PD-L1 combined positive score (CPS) test, was only 57.1%. Most interestingly, the developed method is especially suitable for those patients receiving false negative results in the CPS test. The proposed Fe3O4@SiO2-ILI-01@Tim4 is a highly efficient and robust technique showing great potential in high throughput and noninvasive exosomal PD-L1 detection for accurately predicting immunotherapy efficacy.


Nucleocytoplasmic shuttling of the La motif-containing protein Sro9 might link its nuclear and cytoplasmic functions.
Diverse steps in gene expression are tightly coupled. Curiously, the La-motif-containing protein Sro9 has been shown to play a role in transcription and translation. Here, we show that Sro9 interacts with nuclear and cytoplasmic protein complexes involved in gene expression. In addition, Sro9 shuttles between nucleus and cytoplasm and is exported from the nucleus in an mRNA export-dependent manner. Importantly, Sro9 is recruited to transcribed genes. However, whole genome expression analysis shows that loss of Sro9 function does not greatly change the level of specific transcripts indicating that Sro9 does not markedly affect their synthesis and/or stability. Taken together, Sro9 might bind to the mRNP already during transcription and accompany the mature mRNP to the cytoplasm where it modulates translation of the mRNA.


Lipidome remodeling in response to nutrient replenishment requires the tRNA modifier Deg1/Pus3 in yeast.
In the yeast Saccharomyces cerevisiae, the absence of the pseudouridine synthase Pus3/Deg1, which modifies tRNA positions 38 and 39, results in increased lipid droplet (LD) content and translational defects. In addition, starvation-like transcriptome alterations and induced protein aggregation were observed. In this study, we show that the deg1 mutant increases specific misreading errors. This could lead to altered expression of the main regulators of neutral lipid synthesis which are the acetyl-CoA carboxylase (Acc1), an enzyme that catalyzes a key step in fatty acid synthesis, and its regulator, the Snf1/AMPK kinase. We demonstrate that upregulation of the neutral lipid content of LD in the deg1 mutant is achieved by a mechanism operating in parallel to the known Snf1/AMPK kinase-dependent phosphoregulation of Acc1. While in wild-type cells removal of the regulatory phosphorylation site (Ser-1157) in Acc1 results in strong upregulation of triacylglycerol (TG), but not steryl esters (SE), the deg1 mutation more specifically upregulates SE levels. In order to elucidate if other lipid species are affected, we compared the lipidomes of wild type and deg1 mutants, revealing multiple altered lipid species. In particular, in the exponential phase of growth, the deg1 mutant shows a reduction in the pool of phospholipids, indicating a compromised capacity to mobilize acyl-CoA from storage lipids. We conclude that Deg1 plays a key role in the coordination of lipid storage and mobilization, which in turn influences lipid homeostasis. The lipidomic effects in the deg1 mutant may be indirect outcomes of the activation of various stress responses resulting from protein aggregation.


Ability of some Black Sea organisms to accumulate fission products.
The coefficients of accumulation of strontium-90, cesium-137, and cerium-144 in seaweeds, eelgrass, actinia, mollusks, and crustaceans are presented. The discharge of strontium-90 into sea water from decomposing seaweed and the retention and additional absorption of cesium-137 and cerium-144 onto organic debris is discussed. Some observations are made about the ability of these elements to diffuse into sea water and about the relative hazard to man from strontium-90 and cerium-144 in marine life.


Voltage dependence and stability of the gating kinetics of the fast chloride channel from rat skeletal muscle.
1. The voltage dependence and stability of the gating kinetics of the fast Cl-channel in excised patches of membrane from cultured rat skeletal muscle were studied with the patch clamp technique. Up to 10(6) open and shut intervals were analysed from each of five different patches containing a single fast Cl-channel. 2. To test for kinetic stability, plots of the mean durations of consecutive groups of 5-500 open and shut intervals were examined at each voltage. After excluding infrequent entries into both an apparent subconductance state and a long-lived (inactive) shut state, there were no abrupt and sustained changes in the moving means, indicating the absence of obvious shifts to other kinetic modes. The moving means did, however, fluctuate about the overall mean. 3. A comparison of experimental and simulated data indicated that most, but not all, of the fluctuation in the moving means was due to the stochastic variation inherent in the gating process. The fluctuation not accounted for by stochastic variation was mainly expressed as a slow, low-amplitude, component of drift about the mean. This slow component was unlikely to have arisen from measurement errors. 4. To examine whether the slow drift reflected detectable changes in kinetic modes, the data were divided into consecutive groups of 50,000 intervals. The exponential components describing the distributions were remarkably similar among the different groups, with stochastic variation accounting for most of the observed differences. This finding implies a single kinetic mode throughout the experiment. Thus, any changes in channel activity associated with the slow drift would have little effect on the analysis of gating kinetics presented here. 5. Depolarization increased channel open probability, Popen, for all five channels. This increase had a voltage sensitivity of 17 +/- 4 mV per e-fold change (effective gating charge of 1.6 +/- 0.32 electronic charges at 23 degrees C). Popen was 0.5 at -31 +/- 4 mV. 6. The depolarization-induced increase in Popen typically arose from a decrease in the mean shut time (19 +/- 4 mV per e-fold change; effective gating charge of 1.3 +/- 0.3 at 23 degrees C) and an increase in the mean open time (109 +/- 61 mV per e-fold change; effective gating charge of -0.24 +/- 0.13). 7. Neither plots of Popen versus voltage nor plots of the mean open and mean shut time versus voltage were completely described by a single Boltzmann distribution, suggesting multiple voltage-sensitive steps in channel gating.(ABSTRACT TRUNCATED AT 400 WORDS)


Comparisons of light traps for monitoring adult Culicoides variipennis (Diptera: Ceratopogonidae).
From 1984 to 1986 tests were conducted to evaluate 4 trap types (ABADRL baffle trap, New Jersey trap, CDC trap, and updraft trap) with or without an incandescent light source, a suction fan or dry ice (as a source of CO2) for sampling adult Culicoides variipennis. The fewest flies were caught in the CDC and updraft traps. The New Jersey trap with suction, a strong light source and dry ice caught most flies, due to a significant increase in nulliparous females. The same New Jersey trap without dry ice caught the highest proportion of flies that had taken at least one bloodmeal. This trap would therefore be recommended for use in trapping for virus assay under epizootic conditions.


In vitro and in vivo anti-primary effusion lymphoma activities of fucoidan extracted from Cladosiphon okamuranus Tokida.
Primary effusion lymphoma (PEL) caused by Kaposi's sarcoma-associated herpesvirus (KSHV) is characterized by lymphomatous effusion in body cavities and poor prognosis. There is still no effective treatment for PEL. Fucoidan, a major sulfated polysaccharide isolated from brown seaweeds, has an attractive array of bioactivities such as cancer inhibition. However, the effects of fucoidan on PEL cells remain unclear. We investigated the anti-PEL effects of fucoidan obtained from Cladosiphon okamuranus Tokida cultivated in Okinawa. Fucoidan dose-dependently inhibited the proliferation of KSHV-infected PEL cell lines, and provoked G1 cell cycle arrest, which was accompanied by CDK4 and CDK6 downregulation. Fucoidan also induced apoptosis of PEL cells through caspase-3, -8 and -9 activation; this occurred partly through the downregulation of anti-apoptotic Bcl-xL, Mcl-1 and XIAP proteins. Fucoidan also suppressed nuclear factor-κB, activator protein-1 (AP-1), and T-lymphokine-activated killer cell-originated protein kinase (TOPK) signaling pathways through inhibition of phosphorylation of IκBα and TOPK, and the expression of AP-1 family proteins, JunB and JunD. Oral administration of fucoidan effectively inhibited the development of PEL cells and ascites in a xenograft SCID mouse model, with minimal serious adverse effects. Notably, native fucoidan exhibited a more efficient anti-PEL effect than nanoparticle fucoidan. These preclinical findings highlight the anti-PEL actions of fucoidan, suggesting it could be potentially useful for the prevention and treatment of PEL.


Unbalanced bone turnover in children with beta-thalassemia.
The life expectancy of patients with thalassemia has greatly improved over the last decade as a result of regular transfusions and increased compliance with iron chelation therapy, however, this improvement is often accompanied by a series of serious complications including osteopenia and osteoporosis. The pathogenesis of these skeletal disorders is multifactorial which may be due to hormonal deficiency, compromised nutritional status, bone marrow expansion due to erythroid hyperplasia, increased iron stores or desferrioxamine toxicity. The non invasive assessment of bone turnover has markedly improved with the development of specific and sensitive markers of bone formation. The aim of this work is to assess the value of bone formation markers in patients with beta-thalassemia. To achieve this goal, 36 patients with thalassemia were recruited in this study. There were 20 males (56.6%) and 16 females (44.4%) and their ages ranged from 3 to 18 years. A control group of 20 apparently healthy subjects of matched age and sex was used. The patients were selected from the outpatient clinic and inpatients of the Hematology/Oncology Unit of Mansoura University Children's Hospital (MUCH). The selected subjects were subjected to thorough history taking, clinical examination, radiological evaluation and laboratory investigations in the form of: complete blood count, serum iron, serum ferritin, total iron binding capacity, serum calcium, serum phosphorus and estimation of bone formation markers as alkaline phosphatase and osteocalcin. The results were as follows: serum calcium level was within normal range and showed no statistical significance (p = 0.176) when compared to the control group, while serum phosphorus level was significantly higher in thalassemic patients than the controls (p = 0.002); this may reflect hypoparathyroidism. Analysis of the level of bone formation markers showed serum alkaline phosphatase levels slightly higher in patients than controls but not significant (p = 0.055), and this elevation can be referred to associated liver disease in these patients. On the other hand, osteocalcin level was significantly lower in patients than controls (p = 0.011), and this may be due to osteoblast poisoning by iron overload. In conclusion, thalassemic patients have unbalanced bone turnover between the bone formation and resorption markers and this is evidenced by non significant changes or decreased levels of bone formation markers, while bone resorption is an active process.


Short-term exposure alters adult listeners' perception of segmental phonotactics.
This study evaluates the malleability of adults' perception of probabilistic phonotactic (biphone) probabilities, building on a body of literature on statistical phonotactic learning. It was first replicated that listeners categorize phonetic continua as sounds that create higher-probability sequences in their native language. Listeners were also exposed to skewed distributions of biphone contexts, which resulted in the enhancement or reversal of these effects. Thus, listeners dynamically update biphone probabilities (BPs) and bring this to bear on perception of ambiguous acoustic information. These effects can override long-term BP effects rooted in native language experience.


High Quality QCD Axion at Gravitational Wave Observatories.
The axion solution to the strong CP problem is delicately sensitive to Peccei-Quinn breaking contributions that are misaligned with respect to QCD instantons. Heavy QCD axion models are appealing because they avoid this so-called quality problem. We show that generic realizations of this framework can be probed by the LIGO-Virgo-KAGRA interferometers, through the stochastic gravitational wave (GW) signal sourced by the long-lived axionic string-domain wall network and by upcoming measurements of the neutron and proton electric dipole moments. Additionally, we provide predictions for searches at future GW observatories, which will further explore the parameter space of heavy QCD axion models.


[Diagnosis and therapeutic options in chronic idiopathic intestinal pseudo-obstruction: review of 16 cases].
Chronic idiopathic intestinal pseudo-obstruction (CIPS) is a rare condition in which there is a defective motility of the gastrointestinal tract of unknown cause leading to repeated bouts of intestinal obstruction without organic explanation. This syndrome groups several ill-defined varieties of motor disorders that can sometimes be classified according to the presence of familial incidence and to the presence of muscular or nervous lesions. Nevertheless, a considerable proportion of cases cannot be ascribed to either type. CIPS is a very difficult challenge for pediatric surgeons because our role is never curative and because when we are involved in it is usually as a result of a false diagnosis. We present herein the experience of 2 Pediatric Surgery Departments in this entity. In the last 30 years we have been involved in the management of 16 children with CIPS. Male-to-female ratio was 5:11 and all but 3 patients had symptoms before 6 months of life. Thirteen had abdominal distension, 10 maldevelopment, 9 recurrent bouts of intestinal obstruction, 8 chronic diarrhea, 7 vomiting, 2 dysphagia and 2 constipation. Seven out of the 16 had urinary tract involvement and in three prenatal diagnosis of megacysts was made. The mean delayed time for diagnosis was 3.08 years. Esophageal or antroduodenal manometry was performed in 8 patients and it was abnormal in 7. Histologic and histochemical samples were available in 8 patients, but only in 4 was enough to make a diagnosis of myopathy. Twelve patients underwent 41 surgical procedures. Three are currently included in a program of home parenteral nutrition. Only three have died, and the mean age of the survivors is 13.9 years. In most of the patients with CIPS surgery is only useful for nutritional purposes, for diversion procedures or for intestinal transplantation in extreme cases. Every effort should be made to avoid unnecessary explorations, misdiagnosis and delay in the identification of the syndrome.


Insulin binding, internalization, and insulin receptor regulation in fibroblasts from type II, non-insulin-dependent diabetic subjects.
The ability of insulin to bind, internalize, and regulate its own receptor was investigated in cultured human fibroblasts obtained from 8 normal subjects and 8 patients with type II, non-insulin-dependent diabetes mellitus (NIDDM). The ability of the cells from the two groups to bind insulin was the same, and Scatchard analysis demonstrated identical curvilinear plots. When cells were incubated at 37 degrees C with the lysosomotropic agent, chloroquine, and 125I-insulin, the drug led to a marked, but comparable, increase in cell-associated radioactivity in both control and diabetic fibroblasts (236 and 245% increase, respectively). Insulin pretreatment leads to a loss of insulin receptors in cultured human fibroblasts and preincubation with insulin led to a comparable dose-dependent decrease in subsequent insulin binding in both normal and diabetic fibroblasts. Scatchard analysis demonstrated that this decrease in binding was entirely due to a decrease in receptor number with no change in receptor affinity. These data demonstrate normal insulin binding, insulin internalization, and insulin-mediated receptor loss in fibroblasts from patients with with NIDDM, and these cells are several generations removed form the in vivo milieu. Thus, these results provide direct evidence that the well-known decrease in insulin binding in freshly isolated cells from patients with NIDDM, and these cells are several generations removed from the in vivo milieu. Thus, these results provide direct evidence that the well-known decrease in insulin binding in freshly isolated cells form patients with NIDDM is a reflection of environmental factors rather than an intrinsic (genetic) cellular abnormality.


The cost-utility of cinacalcet in addition to standard care compared to standard care alone for secondary hyperparathyroidism in end-stage renal disease: a UK perspective.
Secondary hyperparathyroidism (SHPT) is a common side effect of end-stage renal disease (ESRD) and is associated with increased risk of fracture and cardiovascular events (CV). Current standard treatment includes dietary control, phosphate binders and vitamin D. However, many patients do not have their parathyroid hormone (PTH), calcium and phosphate levels controlled by this regimen. Cinacalcet is the first of a new class of calcimimetic drugs which suppress PTH production. Although there is convincing evidence of the impact of cinacalcet on serum biomarkers, the long-term clinical implications of treatment are less clear. The aim of this study is to estimate the cost-utility of cinacalcet as an addition to standard treatment of SHPT compared with standard treatment alone. A Markov model was developed to estimate the incremental cost-utility of cinacalcet. Uncertainty was explored through extensive sensitivity analysis. Compared with standard treatment, cinacalcet incurs average additional lifetime costs of pound21,167 per person and confers an additional 0.34 quality adjusted life years, resulting in an incremental cost-effectiveness ratio of pound61,890 (approximately euro89,000) per quality-adjusted life-year (QALY). Extensive one-way sensitivity analysis showed that cinacalcet was only likely to be considered cost-effective if the relative risk of mortality for people with very high levels of PTH was 2.2 compared with people whose PTH reached target levels, or if drug costs were considerably reduced. Probabilistic sensitivity analysis showed cinacalcet was very unlikely to be cost-effective at usual levels of willingness to pay in the National Health Service (NHS). Unless the cost of cinacalcet is considerably reduced, it is unlikely to be considered a cost-effective treatment for people with SHPT.


Neuroanatomy influences speciation rates among anurans.
Evolutionary divergence among animal courtship signals is an important component of the speciation process. In anurans, the preferential response of females to the mating call of conspecific males often maintains reproductive isolation among populations. Much of the information in the call is initially processed in the inner ear, and there is considerable variation in the structure of this organ among lineages of frogs. This variation is responsible for differences in the frequency range to which frog species are sensitive and thus influences the frequency range over which mating calls can effectively diverge. Data suggest that this influence of neuroanatomy on mating call divergence is partly responsible for the different rates of speciation among lineages of frogs.


Differential modulation of neural network and pacemaker activity underlying eupnea and sigh-breathing activities.
Many networks generate distinct rhythms with multiple frequency and amplitude characteristics. The respiratory network in the pre-Bötzinger complex (pre-Böt) generates both the low-frequency, large-amplitude sigh rhythm and a faster, smaller-amplitude eupneic rhythm. Could the same set of pacemakers generate both rhythms? Here we used an in vitro respiratory brainslice preparation. We describe a subset of synaptically isolated pacemakers that spontaneously generate two distinct bursting patterns. These two patterns resemble network activity including sigh-like bursts that occur at low frequencies and have large amplitudes and eupneic-like bursts with higher frequency and smaller amplitudes. Cholinergic neuromodulation altered the network and pacemaker bursting: fictive sigh frequency is increased dramatically, whereas fictive eupneic frequency is drastically lowered. The data suggest that timing and amplitude characteristics of fictive eupneic and sigh rhythms are set by the same set of pacemakers that are tuned by changes in the neuromodulatory state.


Outcomes of Primary Total Knee Arthroplasty Following Septic Arthritis of the Native Knee: A Case-Control Study.
Septic arthritis of the native knee often results in irreversible joint damage leading to the need for total knee arthroplasty (TKA). The purpose of the present study was to examine the intermediate-term risk of periprosthetic joint infection (PJI), aseptic revision, and reoperation following primary TKA in patients with a history of septic arthritis of the native knee as compared with primary TKA performed for the treatment of osteoarthritis. We retrospectively identified 215 primary TKAs performed from 1971 to 2016 at a single institution in patients with a history of septic arthritis of the native knee. Each case was matched 1:1 based on age, sex, body mass index (BMI), and surgical year to a TKA for osteoarthritis. The mean age and BMI were 63 years and 30 kg/m2, respectively. The mean duration of follow-up was 9 years. Survivorships free of PJI at 10 years were 90% and 99% for the septic arthritis and osteoarthritis groups, respectively (hazard ratio [HR] = 6.1; p < 0.01). Ten-year survivorships free of any aseptic revisions were 83% and 93% (HR = 2.5; p < 0.01), and survivorships free of any reoperation were 61% and 84% (HR = 2.9; p < 0.01) for the septic arthritis and osteoarthritis groups, respectively. In addition, as time from the diagnosis of native knee septic arthritis to TKA increased, the relative risk of subsequent infection decreased. Preoperative and 2-year postoperative Knee Society scores were similar between the groups (p = 0.16 and p = 0.19, respectively). There was a 6.1-fold increased risk of PJI in patients undergoing TKA with a history of native knee septic arthritis when compared with controls undergoing TKA for the treatment of osteoarthritis, with a cumulative incidence of 9% at 10 years. Subgroup analysis of the septic arthritis cohort revealed a higher risk of any infection in patients who underwent TKA within the first 5 to 7 years after the diagnosis of septic arthritis as compared with those with a greater duration. Moreover, the 10-year rates of survival free of aseptic revision, any revision, and any reoperation were significantly worse in the native knee septic arthritis cohort. Prognostic Level III. See Instructions for Authors for a complete description of levels of evidence.


The accuracy of sonography for evaluation of internal derangement of the temporomandibular joint in asymptomatic elementary school children: comparison with MR and CT.
In order to clarify the incidence and evolution of disk displacement in the temporomandibular joint (TMJ) in children, we performed a longitudinal analysis in 18 subjects. Some investigators have suggested that sonography can provide information about the articular disk position of the TMJ. The purpose of this study was to determine the diagnostic accuracy of sonography for revealing internal derangement of the TMJ in elementary school children compared with our standard of reference, MR imaging and helical CT. Eighteen children were examined using both sonography and MR imaging or helical CT or both. The sonographically revealed distance between the articular capsule and the lateral surface of the mandibular condyle was measured and compared with that obtained by MR or helical CT scanning. Compared with our MR/CT standard of reference, sonography revealed a sensitivity of 83%, a specificity of 96%, and an accuracy of 92% for identifying disk displacement (defined as a distance of 4 mm or more between the articular capsule and the lateral surface of the mandibular condyle). Although sonography's sensitivity, specificity, and accuracy for the diagnosis of disk displacement were slightly inferior to those of MR or helical CT, we assert it is a useful imaging method for longitudinal investigations of a large group of elementary school children. Internal derangement of the TMJ should be suspected if sonography reveals a distance between the articular capsule and the lateral surface of the mandibular condyle of 4 mm or more.


Synchronous triple carcinoma of the colon and rectum.
Synchronous multiple colorectal cancers are defined as multiple malignant colorectal tumors that occur simultaneously. All tumors are distant from each other, and none are the result of metastasis from other tumors. Here, we present a case of a 79-year-old man who was admitted to our hospital because of a 3-month history of abdominal pain associated with anemia, loss of appetite, and body weight loss. The patient did not have a family history of cancer. Computed tomography revealed bowel wall thickness and mesentery inflammation at the hepatic flexure of the colon and cecum. Colonoscopy revealed a tumor located 10 cm from the anal verge. Colonoscopic examination of the large bowel was not possible because of bowel obstruction due to the rectal tumor. Synchronous triple adenocarcinoma of the colon and rectum was confirmed by pathologic examination. The tumor was surgically resected by two-segment resection of the colon, low anterior resection, and right hemicolectomy. We used intraoperative colonoscopy to confirm that there were no other lesions after the resection of the three tumors. To the best of our knowledge, this is the first case of synchronous triple carcinoma of the colon and rectum in Taiwan. We consider that comprehensive preoperative study, extensive intraoperative exploration, and radical resection can increase the survival rate of patients with synchronous multiple colorectal cancers.


Collagen: finding a solution for the source.
Extracellular matrix (ECM)-based scaffolds, through their inherent bioactivity and molecular recognition signals, provide the ideal substrate for tissue engineering and regenerative applications. Collagen, the most abundant ECM protein, has proven itself to be a very versatile material with applications in many fields, including the leather and food industries, cosmetics, drug delivery, and tissue engineering. However, doubts persist about the optimal source of collagen for tissue engineering applications, given possible immunogenicity and disease transmission associated with animal sources and reduced bioactivity and availability of recombinant technologies. In this special edition, an attempt is made to elucidate the advantages of plant-derived human recombinant collagen and its applications in tissue engineering, particularly skin and wound healing. While results are promising, the widespread use of animal-derived collagen means that recombinant technologies may find applications in niche areas.


Requirement of male-specific dosage compensation in Drosophila females--implications of early X chromosome gene expression.
Dosage compensation equates between the sexes the gene dose of sex chromosomes that carry substantially different gene content. In Drosophila, the single male X chromosome is hypertranscribed by approximately two-fold to effect this correction. The key genes are male lethal and appear not to be required in females, or affect their viability. Here, we show these male lethals do in fact have a role in females, and they participate in the very process which will eventually shut down their function--female determination. We find the male dosage compensation complex is required for upregulating transcription of the sex determination master switch, Sex-lethal, an X-linked gene which is specifically activated in females in response to their two X chromosomes. The levels of some X-linked genes are also affected, and some of these genes are used in the process of counting the number of X chromosomes early in development. Our data suggest that before the female state is set, the ground state is male and female X chromosome expression is elevated. Females thus utilize the male dosage compensation process to amplify the signal which determines their fate.


Tobacco Smoking Behaviors in Cancer Survivors: The Mediation Effect of Personality and Emotional Intelligence.
The smoking behaviour of patients following a cancer diagnosis is a critical risk factor for several physical diseases; it can increase the risk of second primary tumors and lower cancer treatment efficacy. Despite this, a great number of survivors continue to smoke after the diagnosis. This observational, cross-sectional on-line study aimed to assess the relationship between the impact of cancer diagnosis on survivors and their smoking behavior, and whether emotional intelligence and personality might mediate this relationship. Ninety-four Italian survivors completed a set of questionnaires: Big Five Inventory; Fagerström Test for Nicotine Dependence; Impact of Event Scale; Brief Emotional Intelligence Scale. The results obtained from the mediation analyses highlighted that the indirect effect on the relationship between the psychological impact of the diagnosis and smoking behaviors was partially mediated by neuroticism (Intrusion: 95% CI [0.00; 0.11]; Avoidance: 95% CI [0.00; 0.18]). Additionally, the data suggested that the relationship between the psychological impact of the diagnosis and smoking behaviors was partially mediated by the utilization of emotions dimension of emotional intelligence (Intrusion: 95% CI [0.00; 0.10]; Avoidance: 95% CI [0.00; 0.22]). Overall, this study suggests the importance of designing interventions to support smoking interruption based on the "mapping" of individual needs and emotional regulation strategies.


Evaluation of the Incremental Value of a Coronary Artery Calcium Score Beyond Traditional Cardiovascular Risk Assessment: A Systematic Review and Meta-analysis.
Coronary artery calcium scores (CACS) are used to help assess patients' cardiovascular status and risk. However, their best use in risk assessment beyond traditional cardiovascular factors in primary prevention is uncertain. To find, assess, and synthesize all cohort studies that assessed the incremental gain from the addition of a CACS to a standard cardiovascular disease (CVD) risk calculator (or CVD risk factors for a standard calculator), that is, comparing CVD risk score plus CACS with CVD risk score alone. Eligible studies needed to be cohort studies in primary prevention populations that used 1 of the CVD risk calculators recommended by national guidelines (Framingham Risk Score, QRISK, pooled cohort equation, NZ PREDICT, NORRISK, or SCORE) and assessed and reported incremental discrimination with CACS for estimating the risk of a future cardiovascular event. From 2772 records screened, 6 eligible cohort studies were identified (with 1043 CVD events in 17 961 unique participants) from the US (n = 3), the Netherlands (n = 1), Germany (n = 1), and South Korea (n = 1). Studies varied in size from 470 to 5185 participants (range of mean [SD] ages, 50 [10] to 75.1 [7.3] years; 38.4%-59.4% were women). The C statistic for the CVD risk models without CACS ranged from 0.693 (95% CI, 0.661-0.726) to 0.80. The pooled gain in C statistic from adding CACS was 0.036 (95% CI, 0.020-0.052). Among participants classified as being at low risk by the risk score and reclassified as at intermediate or high risk by CACS, 85.5% (65 of 76) to 96.4% (349 of 362) did not have a CVD event during follow-up (range, 5.1-10.0 years). Among participants classified as being at high risk by the risk score and reclassified as being at low risk by CACS, 91.4% (202 of 221) to 99.2% (502 of 506) did not have a CVD event during follow-up. This systematic review and meta-analysis found that the CACS appears to add some further discrimination to the traditional CVD risk assessment equations used in these studies, which appears to be relatively consistent across studies. However, the modest gain may often be outweighed by costs, rates of incidental findings, and radiation risks. Although the CACS may have a role for refining risk assessment in selected patients, which patients would benefit remains unclear. At present, no evidence suggests that adding CACS to traditional risk scores provides clinical benefit.


Loss of Nrf2 in mice evokes a congenital intrahepatic shunt that alters hepatic oxygen and protein expression gradients and toxicity.
The transcription factor Nrf2 (Nfe2l2 nuclear factor, erythroid 2-like 2) regulates gene expression directly, controlling pharmacological and toxicological responses. These processes may also be influenced by the structure of the hepatic vasculature, which distributes blood flow through compartmentalized microenvironments to maintain organismal stability. Castings of the hepatic portal vasculature of albino C57BL/6J but not ICR Nrf2(-/-) mice revealed a congenital intrahepatic shunt that was present in two thirds of Nrf2-disrupted mice. This shunt directly connected the portal vein to the inferior vena cava and displayed characteristics of a patent ductus venosus. Immunohistochemistry revealed that Nrf2(-/-) mice with an intrahepatic shunt manifest changes to hepatic oxygen and protein expression gradients when compared with wild-type (WT) and non-shunted Nrf2(-/-) mice. Centrilobular hypoxia found in WT and Nrf2(-/-) mice without shunts was reduced in Nrf2(-/-) livers with a shunt. Hepatic protein expression of phosphoenolpyruvate carboxykinase (Pepck), normally confined to the periportal zone, exhibited both periportal and centrilobular zonal expression in livers from Nrf2(-/-) mice with an intrahepatic shunt. Centrilobular expression of Cytochrome P450 2E1 (Cyp2e1) was diminished in shunted Nrf2(-/-) livers compared with WT and Nrf2(-/-) livers without shunts. The intrahepatic shunt in Nrf2(-/-) mice was further found to diminish acetaminophen hepatoxicity compared with WT and Nrf2(-/-) non-shunted mice following a 6 h challenge with 250 mg/kg acetaminophen. The presence of an intrahepatic shunt influences several physiological and pathophysiological properties of Nrf2(-/-) mice through changes in blood flow, hepatic oxygenation, and protein expression that extent beyond loss of canonical transactivation of Nrf2 target genes.


Determination of affected risk factors on time to recurrence and death in patients with postoperative gastric cancer using copula function.
In survival studies when the event times are dependent, performing of the analysis by using of methods based on independent assumption, leads to biased. In this paper, using copula function and considering the dependence structure between the event times, a parametric joint distribution has made fitting to the events, and the effective factors on each of these events would be determined. This retrospective cohort study was conducted from March 2003 to March 2007. The data collected from 256 patients with gastric cancer who underwent surgery and that the event time of the two outcomes of death and recurrence for them was recorded. Akaike Information Criterion (AIC) was used to determine of suitable parametric models. Moreover, applying copula function with regard to the relationships between the events, the effect of the risk factors of each of the two outcomes was determined. The data analysis was done using R2.12.1 software. According to the AIC criterion, the Weibull distribution had the best fitting in both of the event times. The median times for recurrence and survival of the patients were estimated 20.2 and 28.1 months respectively. Furthermore, with a fitting of Weibull distribution to the two event times using Clayton copula function, the variables of gender, tumor size and tumor pathological stage on survival, and tumor size and tumor pathological stage on recurrence were significant (P<0.001). Applying copula function for determining specific risk factors of the semi-competing events produces suitable results opposite the common methods which are based on independent assumption of the events.


Impact of Climate Change, Food Insecurity, and COVID-19 on the Health of Neonates and Children: A Narrative Review.
Climate change, food insecurity, and epidemics affect all population sub-groups. This article reviews the current evidence on the relationships between climate change, food insecurity, and the COVID-19 pandemic in the context of newborn and child health. The authors searched Medline, PsycINFO, Cumulative Index to Nursing and Allied Health Literature (CINAHL), and Scopus databases using a structured approach. Food insecurity, particularly from the lack of food access and affordability, increased amidst the COVID-19 pandemic. Factors such as nationwide lockdowns, increased unemployment and financial instability, and school closures precipitated food insecurity. Children born to immigrant parents, belonging to racial and ethnic minority groupsor low-income families, and those who were Autistic were highly vulnerable. Climate change also contributes to food insecurity, with increased susceptibility among neonates and children compared to adults. There is a need for further research on the relationships between climate-linked exposures and COVID-19 transmission. Multisectoral collaborations and multilevel interventions are necessary to mobilize local and national resources for mitigating and preventing the synergistic effects of the three concurrent crises. The evidence-informed discourse on this topic can help in improved preparedness and response for future outbreaks and epidemics. The policy interventions for newborn and child survival need to factor in climate change, food insecurity, and emerging diseases.


Prospects of Graphene-hBN Heterostructure Nanogap for DNA Sequencing.
Recent advances in solid-state nano-device-based DNA sequencing are at the helm of the development of a new paradigm, commonly referred to as personalized medicines. Paying heed to a timely need for standardizing robust nanodevices for cheap, fast, and scalable DNA detection, in this article, the nanogap formed by the lateral heterostructure of graphene and hexagonal boron nitride (hBN) is explored as a potential architecture. These heterostructures have been realized experimentally, and our study boasts the idea that the passivation of the edge of the graphene electrode with hBN will solve many of practical problems, such as high reactivity of the graphene edge and difficulty in controlled engineering of the graphene edge structure, while retaining the nanogap setup as a useful nanodevice for sensing applications. Employing first-principle density-functional-theory-based nonequilibrium Green's function methods, we identify that the DNA building blocks, nucleobases, uniquely couple with the states of the nanogap, and the resulting induced states can be attributed as leaving a fingerprint of the DNA sequence in the computed current-voltage (I-V) characteristic. Two bias windows are put forward: lower (1-1.2 V) and higher (2.7-3 V), where unique identification of all four bases is possible from the current traces, although higher sensitivity is obtained at the higher voltage window. Our study can be a practical guide for experimentalists toward development of a nanodevice DNA sensor based on graphene-hBN heterostructures.


[Risk management in epidural anesthesia].
Epidural anesthesia has many advantages, including block of surgical stress, postoperative pain management and prevention of postoperative complications. Therefore, we should use epidural anesthesia when indicated. However, patients with preexisting spinal stenosis or lumbar radiculopathy have higher incidence of neurologic complications after epidural anesthesia. Epidural abscesses caused by epidural anesthesia are rare. However, epidural abscesses are serious complications in patients. Knowing the risk factor of epidural abscesses is important to prevent epidural abscesses, and early diagnosis and early treatment are needed when suspected. It is important to have measures for safety in performing epidural anesthesia at every hospital. Recently, we have many anesthetic techniques, including epidural anesthesia, remifentanil infusion, ultrasound-guided peripheral nerve blocks and intravenous PCA. Therefore, we should choose an anesthesia method based on the careful evaluation of the benefit and risk balance for the patient's safety to reduce the incidence of complications.


Investigation of Potential Brain Microbiome in Alzheimer's Disease: Implications of Study Bias.
Dysbiotic microbiota in the gastrointestinal tract promotes and aggravates neurodegenerative disorders. Alzheimer's disease (AD) has been shown to correlate to dysbiotic bacteria and the immune, metabolic, and endocrine abnormalities associated with abnormal gut-brain-axis signaling. Recent reports also indicate that brain dysbacteriosis may play a role in AD pathogenesis. To evaluate the presence and differences of brain-region dependent microbiomes in control and AD subjects and the contribution of study bias. Two independent cohorts of postmortem AD brain samples were collected from separate locations, processed with different extraction protocols and investigated for the presence of bacterial DNA indicative of a brain microbiome with V4 16S next generation sequencing. In both cohorts, few differences between the control and AD groups were observed in terms of alpha and beta diversities, phyla and genera proportions. Independent of study in both AD and control subjects the most abundant phyla were Proteobacteria, Firmicutes, Actinobacteria, and Bacteroidetes. Variations in beta diversity between hippocampal and cerebellum samples were observed indicating an impact of brain region on the presence of microbial DNA. Importantly, differences in alpha and beta diversities between the two independent cohorts were found indicating a significant cohort- and processing-dependent effect on the microbiome. Finally, there were cohort-specific correlations between the gut microbiome and subject demographics indicate that postmortem interval may have a significant impact on brain microbiome determination. Regardless of the study bias, this study concludes that bacterial DNA can be isolated from the human brain suggesting that a brain microbiome may exist; however, more studies are required to understand the variation in AD.


Localization of transforming growth factors beta1 and beta2 and epidermal growth factor in IgA nephropathy.
The localization of transforming growth factor (TGF)-beta1. TGF-beta2 and epidermal growth factor (EGF) was investigated in IgA nephropathy, and was compared with the severity of histological damage (including tubulointerstitial lesions). The enzyme antibody method was used to stain paraffin-embedded sections of renal tissue from 42 patients with IgA nephropathy (19 males and 23 females). There was a significant correlation between glomerular positivity for TGF-beta1 and TGF-beta2 and the severity of histological damage. There was also a significant correlation between positivity for TGF-beta1 and TGF-beta2 in the tubular epithelium and tubulointerstitial lesions. In contrast, there was no relationship between glomerular positivity for EGF and histological damage, although there was a significant correlation between positivity for EGF in the tubular epithelium and tubulointerstitial lesions. These findings suggest that TGF-beta1 and TGF-beta2 may be important in the progression of IgA nephropathy, and that the distribution of EGF may also be a useful marker for the progression of renal damage, including tubulointerstitial lesions.


Role of transgelin-2 in diabetes-associated pancreatic ductal adenocarcinoma.
Pancreatic ductal adenocarcinoma (PDAC) is an aggressive malignancy with poor prognosis. Diabetes is a significant risk factor for PDAC and >50% of PDAC patients have concomitant diabetes. How diabetes influences the initiation and progression of PDAC remains elusive. Here, we show that transgelin-2 is dominantly expressed in PDAC tissues compared with adjacent normal tissues. The high level of transgelin-2 indicates poor survival of patients with PDAC. Remarkably, transgelin-2 expression is correlated with diabetic status. Hyperinsulinemia is frequently observed in type 2 diabetes. Our results indicate that upregulation of transgelin-2 is induced by insulin via sterol regulatory element-binding protein (SREBP)-1-mediated transcription in PDAC cells. Transgelin-2 is a novel target of SREBP-1. Our data support a novel mechanism in diabetes-associated PDAC by which transgelin-2 mediates proliferation of PDAC cells upon insulin stimulation. The insulin/SREBP-1/transgelin-2 network should be further explored as a diagnostic marker or a novel therapeutic target for diabetes-associated PDAC.


[Ultrastructure of squamous epithelium in esophageal dysplasia and squamous cell cancer].
Electron microscopic examination of the oesophageal squamous cell carcinoma revealed the presence in the tumour of both undifferentiated and differentiated squamous cells. Keratinocytes appeared and the signs of keratinization were more pronounced in the cytoplasm of differentiated cells. Dysplastic changes were observed at the periphery of the primary node. Two main variants of dysplasia (dark-cell and clear-cell) were distinguished, this at the ultrastructural level being the reflection of the direction of differentiation and the degree of cell maturity in the dysplastic foci. With the exception of few cases with a cell polymorphism in the foci of a severe dysplasia, dysplastic changes of the squamous epithelium were characterized by a monotonous ultrastructural cell composition. The dysplastic cells were distinguished by a degree of differentiation and high synthetic activity.


LABORATORY EVALUATION OF EFFICACY OF THE LARVICIDE SPINOSAD AGAINST ANOPHELES STEPHENSI IN JIGJIGA, ETHIOPIA.
We report the efficacy of a commercial formulation of the insecticide spinosad against larvae of Anopheles stephensi populations found in the city of Jigjiga, Somali Region, eastern Ethiopia. Batches of 25 larvae (late III to early IV instars) collected from large water storage reservoirs associated with construction sites (the primary An. stephensi larval site in the dry season) were tested under laboratory conditions against each insecticide at a dose recommended by the manufacturer (Natular® G30, 0.02 g/5 liter), following World Health Organization guidelines. Mortality at 24-48 h postexposure was 100%. Results show that spinosad is effective against An. stephensi larvae and suggest that it may be a useful tool as part of larval source management plans aimed at controlling this invasive malaria vector in Ethiopia.


Corn hybrid affects in vitro and in vivo measures of nutrient digestibility in dogs.
Corn is a commonly used ingredient in dry pet foods because there is a stable supply and it is a relatively inexpensive source of nutrients. Corn hybrids are available that are higher in CP and amylose and lower in phytate concentration than conventional hybrids. Approximately 500 mg of high-protein (HP), high-protein, low-phytate (HPLP), and high-amylose (HA) corn were compared with conventional (CONV) corn and amylomaize starch (AM) in triplicate and exposed to pepsin/hydrochloric acid and pancreatin to simulate hydrolytic digestion. Substrate remaining after this was used to determine in vitro colonic fermentation. Organic matter disappearances as a result of hydrolytic digestion were >80% for CONV, HP, and HPLP, whereas HA (60.7%) and AM (43.7%) were lower (P < 0.05). Total digestion (TD) values after hydrolytic digestion and 8 h of fermentation using canine fecal inoculum were greater (P < 0.05) for CONV, HP, and HPLP vs. HA and AM. The residue left after hydrolytic digestion of all substrates was poorly fermented. Five ileal-cannulated dogs were fed each corn hybrid at approximately 31% of the diet in a 5 x5 Latin square design. Dogs fed diets containing HP corn had higher (P < 0.05) ileal OM digestibility (70.3%) and tended (P < 0.10) to have higher DM digestibility (64.6%). Ileal starch digestibilities were lower (P < 0.05) for dogs fed HA (64.0%) and AM (63.0%). Ileal digestibilities of essential (71.2%), nonessential (67.4%), and total (69.0%) AA tended to be higher (P < 0.10) for HP diets compared with CONV (66.4, 62.4, and 64.0%, respectively). Total-tract DM, OM, CP, and GE digestibilities (77, 82, 77, and 84%, on average, respectively) were higher (P < 0.05) for dogs fed CONV, HP, and HPLP than for those fed AM (66.9, 71.6, 72.6, and 76.5%) and HA (60.6, 65.7, 69.7, and 71.5%). Total-tract fat digestibilities were lower (P < 0.05) for dogs fed HA diets (86.6%) than for all other treatments (91.0%, on average). Total-tract starch digestibilities were higher (P < 0.05) for dogs fed CONV, HP, and HPLP (98%, on average) compared with HA (72.8%) and AM (76.5%). No differences were detected among treatments in fecal bifidobacteria, lactobacilli, or Clostridium perfringens concentrations. The experiments demonstrated that HP and HPLP corn had hydrolytic digestion and fermentation characteristics similar to those of CONV corn, whereas HA resulted in similar responses to AM, a well-established resistant starch ingredient.


Diffusion and perfusion weighted magnetic resonance imaging for tumor volume definition in radiotherapy of brain tumors.
Accurate target volume delineation is crucial for the radiotherapy of tumors. Diffusion and perfusion magnetic resonance imaging (MRI) can provide functional information about brain tumors, and they are able to detect tumor volume and physiological changes beyond the lesions shown on conventional MRI. This review examines recent studies that utilized diffusion and perfusion MRI for tumor volume definition in radiotherapy of brain tumors, and it presents the opportunities and challenges in the integration of multimodal functional MRI into clinical practice. The results indicate that specialized and robust post-processing algorithms and tools are needed for the precise alignment of targets on the images, and comprehensive validations with more clinical data are important for the improvement of the correlation between histopathologic results and MRI parameter images.


Epigenetic Alterations in Prescription Opioid Misuse: New Strategies for Precision Pain Management.
Prescription opioids are used for some chronic pain conditions. However, generally, long-term therapy has unwanted side effects which may trigger addiction, overdose, and eventually cause deaths. Opioid addiction and chronic pain conditions have both been associated with evidence of genetic and epigenetic alterations. Despite intense research interest, many questions about the contribution of epigenetic changes to this typology of addiction vulnerability and development remain unanswered. The aim of this review was to summarize the epigenetic modifications detected in specific tissues or brain areas and associated with opioid prescription and misuse in patients who have initiated prescribed opioid management for chronic non-cancer pain. The review considers the effects of opioid exposure on the epigenome in central and peripheral tissues in animal models and human subjects and highlights the mechanisms in which opioid epigenetics may be involved. This will improve our current understanding, provide the basis for targeted, personalized pain management, and thus balance opioid risks and benefits in managing chronic pain.


Genomic sequence and expression of a cloned human carbonyl reductase gene with daunorubicin reductase activity.
Carbonyl reductase (NADPH: secondary-alcohol oxidoreductase; EC 18.104.22.168), a widely distributed NADPH-dependent enzyme considered as both an aldo-keto reductase and a quinone reductase, was cloned from a human liver genomic library and transiently expressed in COS7 cells. The gene contains 3142 bases comprising three exons and two introns. The absence of a CAAT and TATA box and the presence of a GC-rich island are characteristic of many "housekeeping" genes. Transient expression of the genomic gene in COS7 cells using an expression vector containing an SV40 origin of replication resulted in a greater than 50-fold increase in both menadione reductase activity and daunorubicin reductase activity, suggesting that both activities are derived from the same enzyme. Carbonyl reductase mRNA levels reflected enzyme activity levels in the transfected cells. Other parameters, such as pH profile, cofactor requirements, substrates, and inhibitors, were similar to those of carbonyl reductase purified by other investigators. Potential regulatory elements with consensus sequences for two GC boxes and the transcriptional activator protein AP-2 were present upstream of the transcriptional start site. Although the precise role of carbonyl reductase is unknown, the enzyme is involved in drug metabolism and in the reduction of activated carbonyl compounds. Its ability to act as a quinone reductase also implies a potential to modulate oxygen free radicals.


Secretion of RNA-Containing Extracellular Vesicles by the Porcine Whipworm, Trichuris suis.
Trichuris suis is a common parasitic helminth of pigs. As with many other parasites, T. suis ensures its own survival by evading host immune responses, but little is known about how this is achieved. MicroRNAs (miRNA) have been shown to be involved in various immunological processes by post-transcriptional regulation of specific genes, and the potential of using these molecules as biomarkers of disease is currently being examined. It has recently been shown that parasites may secrete extracellular structures such as exosomes and microvesicles, containing proteins and miRNA. The fusion of these structures with host cells has been demonstrated, and a role of exosome-derived miRNA in host gene regulation has been suggested. In the present study, we show that exosome- and microvesicular-like structures are secreted by T. suis L1 larvae and also demonstrate the presence of miRNA-sized RNA inside these structures. A potential role of these molecules in host-parasite interactions is suggested. In addition, an electron-dense layer covering the surface of the larvae was observed, which may play a function in host immune evasion.


A continuous responder algorithm to optimize clinical management of small-cell lung cancer with progastrin-releasing peptide as a simple blood test.
This study aimed to investigate whether changes in progastrin-releasing peptide (ProGRP) levels correlate with treatment response and can be used to optimize clinical management of patients with small-cell lung cancer. Patients with small-cell lung cancer (any stage) receiving chemotherapy were eligible. ProGRP was measured in serum/plasma at baseline and after each chemotherapy cycle using the Elecsys® ProGRP assay (Roche Diagnostics). Treatment response was assessed by computed tomography scan. The primary objective was to examine whether changes in ProGRP levels correlated with computed tomography scan results after two cycles of chemotherapy. The prognostic value of ProGRP among patients receiving first-line chemotherapy was also assessed. Overall, 261 patients from six centers were eligible. Among patients with elevated baseline ProGRP (>100 pg/mL), a ProGRP decline after Cycle 2 was associated with nonprogression (area under the curve: 84%; 95% confidence interval: 72.8-95.1; n = 141). ProGRP changes from baseline to end of Cycle 1 were predictive of response, as determined by computed tomography scan 3 weeks later (area under the curve: 87%; 95% confidence interval: 74.1-99.2; n = 137). This was enhanced by repeat measurements, with a 92% area under the curve (95% confidence interval: 85.3-97.8) among patients with ProGRP data after both Cycles 1 and 2 (n = 123); if a patient experienced a ≥25% decline in ProGRP after Cycle 1, and ProGRP remained stable or decreased after Cycle 2, the probability of finding progression on the interim computed tomography scan at the end of Cycle 2 was almost zero (sensitivity: 100%, specificity: 71%). Both ProGRP levels at baseline and at the end of first-line chemotherapy were prognostic; the latter provided a moderately improved hazard ratio of 2.43 (95% confidence interval: 1.33-4.46; n = 110) versus 1.87 (95% confidence interval: 1.04-3.37; n = 216). In summary, for patients with small-cell lung cancer and elevated baseline ProGRP levels, ProGRP may be a simple, reliable, and repeatable tool for monitoring response to chemotherapy and provide valuable prognostic information.


AML-associated Flt3 kinase domain mutations show signal transduction differences compared with Flt3 ITD mutations.
Activating mutations of Flt3 are found in approximately one third of patients with acute myeloid leukemia (AML) and are an attractive drug target. Two classes of Flt3 mutations occur: internal tandem duplications (ITDs) in the juxtamembrane and point mutations in the tyrosine kinase domain (TKD). We and others have shown that Flt3-ITD induced aberrant signaling including strong activation of signal transducer and activator of transcription 5 (STAT5) and repression of CCAAT/estradiol-binding protein alpha (c/EBPalpha) and Pu.1. Here, we compared the signaling properties of Flt3-ITD versus Flt3-TKD in myeloid progenitor cells. We demonstrate that Flt3-TKD mutations induced autonomous growth of 32D cells in suspension cultures. However, in contrast to Flt3-ITD and similar to wild-type Flt3 (Flt3-WT), Flt3-TKD cannot support colony formation in semisolid media. Also, in contrast to Flt3-ITD, neither Flt3-WT nor Flt3-TKD induced activation or induction of STAT5 target genes. Flt3-TKD also failed to repress c/EBPalpha and Pu.1. No significant differences were observed in receptor autophosphorylation and the phosphorylation of Erk-1 and -2, Akt, and Shc. Importantly, TKD but not ITD mutations were a log power more sensitive toward the tyrosine kinase inhibitor protein kinase C 412 (PKC412) than Flt3-WT. In conclusion, Flt3-ITD and Flt3-TKD mutations display differences in their signaling properties that could have important implications for their transforming capacity and for the design of mutation-specific therapeutic approaches.


Prestroke dementia: characteristics and clinical features in consecutive series of patients.
The prestroke level of cognitive function should be taken into account in order to predict the impact of stroke on the subsequent risk of dementia. The aim of the present study was to investigate the presence and correlates of prestroke dementia (PSD) as well as to identify its clinical features. Premorbid clinical and cognitive features of 158 consecutively recruited patients with a diagnosis of acute cerebrovascular pathology were assessed by interviewing the caregivers using multidimensional assessment. Patients were divided into two groups (PSD group and prestroke nondemented group). Baseline cognitive, functional and behavioral variables and neuroradiological hallmarks (medial temporal lobe atrophy, MTLA) were compared between these two groups. In a logistic regression model, older age (OR 1.05), female gender (OR 2.3), Neuropsychiatric Inventory total score (OR 1.1) and MTLA (OR 1.2) were the variables independently associated with PSD. These findings support the hypothesis that cognitive impairment in patients with stroke may not only be a direct consequence of the acute cerebrovascular event but also a consequence of underlying neurodegenerative pathology.


New reactions in the crotonase superfamily: structure of methylmalonyl CoA decarboxylase from Escherichia coli.
The molecular structure of methylmalonyl CoA decarboxylase (MMCD), a newly defined member of the crotonase superfamily encoded by the Escherichia coli genome, has been solved by X-ray crystallographic analyses to a resolution of 1.85 A for the unliganded form and to a resolution of 2.7 A for a complex with an inert thioether analogue of methylmalonyl CoA. Like two other structurally characterized members of the crotonase superfamily (crotonase and dienoyl CoA isomerase), MMCD is a hexamer (dimer of trimers) with each polypeptide chain composed of two structural motifs. The larger N-terminal domain contains the active site while the smaller C-terminal motif is alpha-helical and involved primarily in trimerization. Unlike the other members of the crotonase superfamily, however, the C-terminal motif is folded back onto the N-terminal domain such that each active site is wholly contained within a single subunit. The carboxylate group of the thioether analogue of methylmalonyl CoA is hydrogen bonded to the peptidic NH group of Gly 110 and the imidazole ring of His 66. From modeling studies, it appears that Tyr 140 is positioned within the active site to participate in the decarboxylation reaction by orienting the carboxylate group of methylmalonyl CoA so that it is orthogonal to the plane of the thioester carbonyl group. Surprisingly, while the active site of MMCD contains Glu 113, which is homologous to the general acid/base Glu 144 in the active site of crotonase, its carboxylate side chain is hydrogen bonded to Arg 86, suggesting that it is not directly involved in catalysis. The new constellation of putative functional groups observed in the active site of MMCD underscores the diversity of function in this superfamily.


Bcl-2 and p53 expression in stage I endometrial carcinoma.
The role of p53 and bcl-2 apoptosis related proteins in the pathogenesis of endometrial cancer remains unclear. We immunohistochemically examined 133 surgically removed tumour specimens from patients with stage I endometrial cancer for p53 oncoprotein nuclear expression and bcl-2 cytoplasmic reactivity. 114/133 (86%) cases were of the endometrioid histological sub-type. A cut-off point of 10% of cells with strong reactivity was used for positivity. Positive p53 expression was observed in 12/133 (9%) and positive bcl-2 in 40/133 (30%) cases. p53 expression was not related to bcl-2 expression. No association of p53 and bcl-2 with depth of myometrial invasion, vascular invasion or histological grade was observed. However, continuous variable analysis revealed a trend of low grade cases to have a higher percentage of bcl-2 positive cells (16.3 + 27% vs. 7.8 + 16%; p = 0.09, unpaired two tailed t-test). Interestingly, a statistically significant association of p53 positivity with age was also observed (p = 0.03). A strong association of high grade with depth of myometrial invasion (p = 0.006) and vascular invasion (p = 0.0001) was also noticed. In addition, the presence of adenomyosis was also associated with low grade (p = 0.01) and absence of vascular invasion (p = 0.02). We conclude that although loss of bcl-2 protein expression and p53 mutant protein nuclear accumulation are early events in the endometrial cancer progression, histological grade and vascular invasion remain the most important factors defining local invasive behaviour of endometrial cancer.


No evidence of "weaponized Title IX" here: An empirical assessment of sexual misconduct reporting, case processing, and outcomes.
Little is known about actual incidents of gender-based violence reported by college students or the campus adjudication process or outcomes of reported cases. Data from Annual Security Reports (ASRs) and Title IX Coordinators was used to examine the context, processes, and outcomes of reported incidents of sexual misconduct (N = 1,054) at institutions of higher education (IHEs) in a Mid-Atlantic state. Results showed that ASRs undercounted incidents of sexual misconduct. Few incidents reported to Title IX Coordinators resulted in a formal Title IX complaint, and fewer still resulted in a finding of responsibility or suspension/expulsion of the responsible student. The primary outcome of reports were victim services, not perpetrator punishments. Significant variability within and between IHE types was also uncovered. Findings suggest that better data collection as well as research on victim engagement in the Title IX complaint process and on sexual misconduct at community colleges and independent IHEs is needed. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Six-Month Community-Based Brisk Walking and Balance Exercise Alleviates Motor Symptoms and Promotes Functions in People with Parkinson's Disease: A Randomized Controlled Trial.
In Parkinson's disease (PD), sustained aerobic exercise is a promising therapy in delaying motor disability. Brisk walking is a moderate intensity aerobic training, which could be translated to community practice at low cost, but its effects on motor symptoms remains unclear. To determine the effectiveness of a six-month brisk walking and balance program in alleviating motor symptoms, and promoting functional, gait, and balance performance in people with PD. Seventy individuals with mild to moderate PD were randomly assigned to a brisk walking (BW) group or an active control (CON) group. BW group received ten 90-minute supervised brisk walking and balance exercise for six months (weeks 1-6: once/week, weeks 7-26: once/month). CON group received upper limb training. Both groups performed 2-3 self-practice sessions weekly. Primary outcome was Movement Disorder Society Unified Parkinson's Disease Rating Scale (MDS-UPDRS) motor score. Secondary outcomes were fast gait speed (FGS), timed-up-and-go (TUG) time, six-minute walk distance (6MWD), and Mini-Balance Evaluation Systems Test (Mini-BEST) score. Sixty-four participants (33 BW/31 CON) completed training. BW group showed greater significant decreases from baseline than CON group in MDS-UPDRS motor score after six weeks (-5.5 vs -1.6, p < 0.001) and 6 months (-6.0 vs -1.4, p < 0.001) of training. BW group also showed greater significant improvement from the baseline than CON group for TUG time, FGS, 6MWD, and mini-BEST score (all p < 0.05). The six-month brisk walking and balance program alleviates motor symptoms, promotes functional and gait performance, walking capacity, and dynamic balance in people with mild to moderate PD.


Superior mesenteric artery syndrome caused by celiac axis compression syndrome: a case report and review of the literature.
Celiac axis compression syndrome (CACS) is a rare entity of mesenteric ischemia, secondary to inadequate blood supply to the intestine, resulting in weight loss because of postprandial abdominal pain. Superior mesenteric artery (SMA) syndrome is an uncommon cause of intestinal obstruction manifesting with epigastric pain, bilious vomiting, and postprandial discomfort. Although the coexistence of both syndromes is very rare and has been reported only in eight patients in the literature, the CACS as a rare etiology of SMA syndrome has not yet been reported. Herein, we describe an uncommon case of SMA syndrome secondary to the CACS. The 27-year-old woman presented with epigastric pain, postprandial vomiting, and rapid body weight loss. The diagnosis of SMA syndrome was made by hypotonic duodenography and multidetector computer tomographic angiography. The CACS was also suspected by multidetector computer tomographic angiography. Surgical intervention was performed and the presence of CACS was confirmed. Her symptoms subsided shortly after operation and she was in good health at 1-year follow-up.


Dentist involvement in care provided by the dental hygienist.
This study investigated a nationwide sample of dental hygienists to determine the extent of dentist involvement during the patient's appointment with the dental hygienist. The American Dental Hygienists' Association identified the need for additional study of dentist involvement and contracted for data to be obtained as part of the ongoing longitudinal study of 1982 dental hygiene graduates. Mail questionnaires were sent to a nationwide cohort of 1,008 dental hygienists who graduated in 1982. Responses were received from 812 subjects, an 83% response rate from subjects with valid addresses. Data indicated that, on the average, the dentist spends 10 minutes with the dental hygienist's patient at the end of the dental hygiene appointment. This time is divided almost equally among talking with the patient, performing an oral examination, and discussing diagnosis and treatment needs.


HIV incidence and risk behaviours after voluntary HIV counselling and testing (VCT) among adults aged 19-35 years living in peri-urban communities around Chiang Mai city in northern Thailand, 1999.
The Thai government began HIV voluntary counselling and testing (VCT) in all provinces in 1992. We evaluated HIV incidence after participants utilized VCT and its effect on risk behaviours among northern Thai adults aged 19-35 years. A total of 924 men and 1,327 women volunteered to participate in the study. Subjects were counseled, interviewed (socio-demographic, HIV risk behaviour, and HIV testing history), and tested for HIV infection at baseline and at 6 months follow-up. All participants were provided group pre-test counselling; HIV test results were provided in individual confidential post-test counselling. Overall, 329 of 391 men and 621 of 669 women who reported a prior HIV test before participating in our study reported negative results at the previous test. Of these, nine men and 13 women tested positive at baseline, giving incidence rates of 1.04 and 0.69 per 100 person years (PY), respectively. Recent risk behaviour was rare; as a result, the effects of VCT on risk behaviours among the study participants could not be determined. The HIV incidence after VCT among this study population is substantial. Studies to investigate factors associated with HIV incidence among VCT clients could provide insights for more effective HIV prevention.


State and trait anxiety in patients affected by nasal polyposis before and after medical treatment.
The present study aimed to establish whether anxiety and depression in nasal polyposis play a role in genesis of the disease, or are a consequence of symptoms. Anxiety levels were evaluated in state and trait forms, and depression, in 30 consecutive patients presenting nasal polyposis before and after effective 7 months' medical treatment with nasal mometasone, loratadine and montelukast. Before and at the end of treatment, patients were asked to fill in the State and Trait Anxiety Inventory and the Zung self-rating depression scale. In 63.15% of patients with high levels of state anxiety before therapy, these were reduced (p < 0.004) after treatment. In 61.9% of patients with high levels of trait anxiety before treatment, these were reduced (p < 0.002) after treatment. There were no significant differences in depression. Anxiety in nasal polyposis is present both as a state and as a trait, and is significantly reduced after effective medical treatment, showing that anxiety is a reversible consequence of nasal polyposis in most cases.


Human olfactory detection of packaged cannabis.
Olfactory detection of cannabis aroma by police officers can be the basis for warrantless searches of motor vehicles in many jurisdictions in the United States. The odor source in these cases is often dried cannabis flower contained in various casual wrappings as well as in more elaborate packaging. Here we investigate whether packaging format alters the detectability of the cannabis. Two cannabis strains and five packaging formats were evaluated. Untrained observers were presented with two containers and asked to identify, based only on smell, the container that held a sample of packaged cannabis (the other container held identical, but empty, packaging material). The results showed that open and casually packaged cannabis was identified with high accuracy, while material packaged in doubly vacuum-sealed plastic was correctly identified at rates no different from chance. The results may help address issues involving the detectability of cannabis aroma in law enforcement and other scenarios.


Characterization of bacteria isolated from wild legumes in the north-western regions of China.
Nodule isolates from 11 species of wild legumes in north-western China were characterized by numerical taxonomy, PCR-based 16S rRNA gene RFLP and sequence analyses, DNA-DNA hybridization, restriction patterns of nodDAB and nifH genes, and symbiotic properties. Based on the results of numerical taxonomy, most of the 35 new isolates were grouped into five clusters (clusters 7, 9, 12, 14 and 15). Clusters 7 and 12 were identified as Mesorhizobium amorphae and Agrobacterium tumefaciens, respectively, based on their high DNA homologies with the reference strains for these species, their 16S rRNA gene analysis and their phenotypic features. Results of 16S rDNA PCR-RFLP analysis showed that cluster 9 belonged to Rhizobium. Clusters 14 and 15 were identified as Mesorhizobium based on their moderately slow-growing, acid-producing characters and the high similarity of their 16S rDNA PCR-RFLP patterns to those of Mesorhizobium species. These two clusters were genomic species distinct from all described species based on analysis of DNA relatedness within this genus. The isolates in cluster 12 (Agrobacterium tumefaciens) failed to nodulate their original host and other selected hosts and they did not hybridize to nif or nod gene probes. The possibility of opportunistic nodulation of these isolates is discussed. Identical restriction patterns were obtained in the nif or nod gene hybridization studies from the three isolates within cluster 15, which were isolated from the same host species. The isolates from different host plants in each of clusters 9 and 14 produced different nodDAB RFLP patterns, but similar nifH RFLP patterns appeared (one band for each isolate). Different patterns were observed among different clusters from both the nod and nif gene hybridization studies. Crossnodulation was recorded among the isolates and the host plants in the same cluster and promiscuous properties were found among some of the hosts tested.


Eosinophil granulocytes account for indoleamine 2,3-dioxygenase-mediated immune escape in human non-small cell lung cancer.
Indoleamine 2,3-dioxygenase (IDO), a catabolizing enzyme of tryptophan, is supposed to play a role in tumor immune escape. Its expression in solid tumors has not yet been well elucidated: IDO can be expressed by the tumor cells themselves, or by ill-defined infiltrating cells, possibly depending on tumor type. We have investigated IDO expression in 25 cases of non-small cell lung cancer (NSCLC). Using histochemistry and immunohistochemistry, we found that IDO was expressed not by tumor cells, but by normal cells infiltrating the peritumoral stroma. These cells were neither macrophages nor dendritic cells, and were identified as eosinophil granulocytes. The amount of IDO-positive eosinophils varied in different cases, ranging from a few cells to more than 50 per field at x200 magnification. IDO protein in NSCLC was enzymatically active. Therefore, at least in NSCLC cases displaying a large amount of these cells in the inflammatory infiltrate, IDO-positive eosinophils could exert an effective immunosuppressive action. On analyzing the 17 patients with adequate follow-up, a significant relationship was found between the amount of IDO-positive infiltrate and overall survival. This finding suggests that the degree of IDO-positive infiltrate could be a prognostic marker in NSCLC.


Mechanism of ammonia excretion in the freshwater leech Nephelopsis obscura: characterization of a primitive Rh protein and effects of high environmental ammonia.
Remarkably little is known about nitrogenous excretion in freshwater invertebrates. In the current study, the nitrogen excretion mechanism in the carnivorous ribbon leech, Nephelopsis obscura, was investigated. Excretion experiments showed that the ribbon leech is ammonotelic, excreting 166.0 ± 8.6 nmol·grams fresh weight (gFW)(-1)·h(-1) ammonia and 14.7 ± 1.9 nmol·gFW(-1)·h(-1) urea. Exposure to high and low pH hampered and enhanced, respectively, ammonia excretion rates, indicating an acid-linked ammonia trapping mechanism across the skin epithelia. Accordingly, compared with body tissues, the skin exhibited elevated mRNA expression levels of a newly identified Rhesus protein and at least in tendency the Na(+)/K(+)-ATPase. Pharmacological experiments and enzyme assays suggested an ammonia excretion mechanism that involves the V-ATPase, Na(+)/K(+)-ATPase, and carbonic anhydrase, but not necessarily a functional microtubule system. Most importantly, functional expression studies of the identified Rh protein cloned from leech skin tissue revealed an ammonia transport capability of this protein when expressed in yeast. The leech Rh-ammonia transporter (NoRhp) is a member of the primitive Rh protein family, which is a sister group to the common ancestor of vertebrate ammonia-transporting Rh proteins. Exposure to high environmental ammonia (HEA) caused a new adjustment of body ammonia, accompanied with a decrease in NoRhp and Na(+)/K(+)-ATPase mRNA levels, but unaltered ammonia excretion rates. To our knowledge, this is only the second comprehensive study regarding the ammonia excretion mechanisms in a freshwater invertebrate, but our results show that basic processes of ammonia excretion appear to also be comparable to those found in freshwater fish, suggesting an early evolution of ionoregulatory mechanisms in freshwater organisms.


Complete primary structure of a human plasma membrane Ca2+ pump.
cDNAs coding for a plasma membrane Ca2+ pump were isolated from a human teratoma library and sequenced. The translated sequence contained 1,220 amino acids with a calculated molecular weight of 134,683. All regions of functional importance known from other ion-transporting ATPases could be identified. The translated sequence also contained, near the carboxyl terminus, the calmodulin-binding domain and two domains which are very rich in glutamic acid and aspartic acid. These two domains resemble calmodulin somewhat and one of them may play a role in the binding of Ca2+. The enzyme also contains domains rich in serine and threonine, one of which has a sequence matching those of good cAMP-dependent protein kinase substrates. The carboxyl-terminal region is important for regulation by calmodulin, proteolysis, and phosphorylation. Near the amino terminus are two domains which are very rich in lysine and glutamic acid, as well as two domains resembling EF hands, one of which also has some resemblance to calmodulin. Comparison of the cloned sequence with peptide sequences from the erythrocyte Ca2+ pump showed that the two proteins have a very high proportion of identical residues but are not 100% identical, indicating that they represent different isozymes.


Human behavioural research applied to the leprosy control programme of Sarawak, Malaysia.
In 1984, in Sarawak, there were a total of 1,099 recorded cases of leprosy for a population of 1.3 million. However, for each case recorded, it is estimated that two others remain undiagnosed as a consequence of the stigmatization associated with leprosy. For the five year period, 1979-1983, an average of 29 new cases were detected each year of which 8.6 (30%) were deformed due to the late stages at which it was being reported. To increase the case-finding rate, human behavioural research was applied to the leprosy control programme so as to develop culture-specific health education packages aimed at self diagnosis and self referral in order to detect the large pool of undiagnosed cases hidden behind the veil of aversion, fear and ignorance. This was achieved through anthropological studies to identify how the various major ethnic groups perceived leprosy and their attitudes towards leprosy. Taking into account these findings, health education packages aimed at adults as well as children were developed for the Chinese as well as the non-Chinese, and consisted of newspaper articles, cartoon tape-slides, cartoon story books and posters.


In vivo human skin barrier modulation by topical application of fatty acids.
The in vivo effects of fatty acids on skin barrier function were assessed by measuring: (i) transepidermal water loss (TEWL), (ii) diffusion lag times for hexyl nicotinate (HN), and (iii) irritant skin response using laser Doppler velocimetry (LDV) in combination with visual scoring. Two classes of fatty acids have been investigated: straight-chain saturated fatty acids (SFA), having 6-12 carbon atoms, and unsaturated fatty acids (UFA): oleic, linoleic, alpha-linolenic and arachidonic acids. It has been reported that these acids can enhance the permeation of various compounds across the skin. After topical and occlusive application as a solution in propylene glycol (PG) for 3 h on the volar arm of human subjects, SFA only caused a slight irritation and increase in TEWL. The diffusion lag times of HN were reduced by the application SFA to the same extent as and not more than by the application of the pure solvent PG. In contrast, the application of UFA caused a significant increase in TEWL and LDV (irritation) responses. The TEWL values after oleic acid application were higher than those observed for the other three acids, while the irritation potential of arachidonic acid was the highest among UFA. As with SFA, sites treated with UFA did not show significantly different lag times of HN diffusion from PC-treated sites. The data suggest that the degree of irritation and the degree of barrier modulation for fatty acids are not necessarily correlated.


Introduction to the Research Symposium on Bilingualism Forum.
The purpose of this introduction is to provide an overview of the articles contained within this research forum of the Journal of Speech, Language, and Hearing Research. Each of these articles is based upon presentations from the 2022 ASHA Research Symposium.


Mouse hepatitis virus 3C-like protease cleaves a 22-kilodalton protein from the open reading frame 1a polyprotein in virus-infected cells and in vitro.
The 3C-like proteinase (3CLpro) of mouse hepatitis virus (MHV) is predicted to cleave at least 11 sites in the 803-kDa gene 1 polyprotein, resulting in maturation of proteinase, polymerase, and helicase proteins. However, most of these cleavage sites have not been experimentally confirmed and the proteins have not been identified in vitro or in virus-infected cells. We used specific antibodies to identify and characterize a 22-kDa protein (p1a-22) expressed from gene 1 in MHV A59-infected DBT cells. Processing of p1a-22 from the polyprotein began immediately after translation, but some processing continued for several hours. Amino-terminal sequencing of p1a-22 purified from MHV-infected cells showed that it was cleaved at a putative 3CLpro cleavage site, Gln_Ser4014 (where the underscore indicates the site of cleavage), that is located between the 3CLpro domain and the end of open reading frame (ORF) 1a. Subclones of this region of gene 1 were used to express polypeptides in vitro that contained one or more 3CLpro cleavage sites, and cleavage of these substrates by recombinant 3CLpro in vitro confirmed that amino-terminal cleavage of p1a-22 occurred at Gln_Ser4014. We demonstrated that the carboxy-terminal cleavage of the p1a-22 protein occurred at Gln_Asn4208, a sequence that had not been predicted as a site for cleavage by MHV 3CLpro. Our results demonstrate the usefulness of recombinant MHV 3CLpro in identifying and confirming cleavage sites within the gene 1 polyprotein. Based on our results, we predict that at least seven mature proteins are processed from the ORF 1a polyprotein by 3CLpro and suggest that additional noncanonical cleavage sites may be used by 3CLpro during processing of the gene 1 polyprotein.


Combination design of drive mechanism considering delay of surgical robot.
Surgical robots have contributed greatly to the development of minimally invasive surgery. However, these robots suffer from issues such as the looseness of wires acting as the robot's drive mechanism, and high maintenance cost. The objective of this study was to derive a combination of driving mechanisms other than wires that satisfies the required positional accuracy of a surgical robot. The robot operation trajectory was obtained using a virtual surgical simulator. Based on the operation trajectory obtained in the suture task experiment, the delay of each joint was virtually simulated, and the robot's position was calculated. The positions of the robot with and without delay were compared, and the drive mechanism combinations that satisfy the required positional accuracy were obtained. Thus, satisfactory accuracy was achieved when the alternative drive mechanism was properly placed.Clinical Relevance- This paper presents an accurate and low-maintenance surgical robot that uses few wires and alternative drive mechanisms.


Drosophila as a model for intractable epilepsy: gilgamesh suppresses seizures in para(bss1) heterozygote flies.
Intractable epilepsies, that is, seizure disorders that do not respond to currently available therapies, are difficult, often tragic, neurological disorders. Na(+) channelopathies have been implicated in some intractable epilepsies, including Dravet syndrome (Dravet 1978), but little progress has been forthcoming in therapeutics. Here we examine a Drosophila model for intractable epilepsy, the Na(+) channel gain-of-function mutant para(bss1) that resembles Dravet syndrome in some aspects (parker et al. 2011a). In particular, we identify second-site mutations that interact with para(bss1), seizure enhancers, and seizure suppressors. We describe one seizure-enhancer mutation named charlatan (chn). The chn gene normally encodes an Neuron-Restrictive Silencer Factor/RE1-Silencing Transcription factor transcriptional repressor of neuronal-specific genes. We identify a second-site seizure-suppressor mutation, gilgamesh (gish), that reduces the severity of several seizure-like phenotypes of para(bss1)/+ heterozygotes. The gish gene normally encodes the Drosophila ortholog of casein kinase CK1g3, a member of the CK1 family of serine-threonine kinases. We suggest that CK1g3 is an unexpected but promising new target for seizure therapeutics.


Traumatic haemothorax--experience of a protocol for rapid turnover in 1,845 cases.
Incomplete or prolonged drainage of the pleural cavity for haemothorax may lead to the development of empyema, with long-term morbidity. Using a protocol based on vigorous physiotherapy and early withdrawal of the thoracostomy tube (average drainage time 27.1 hours), hospital stay in 1,845 patients with traumatic haemothorax was 48 hours or less in 81.8% of patients. In all, 152 haemothorax patients (8.1%) required either early or late thoracotomy, 46 of which were for associated cardiac injury. Prophylactic antibiotics were not given routinely. Severe complications occurred in 40 patients (2.2%); 15 developed empyema (0.8%) and 25 died (1.4%), mainly from ongoing haemorrhage. The early identification of patients needing operative or other intervention minimises the hospital stay and complications associated with residual blood in the pleural cavity. This study shows that the short-period drainage protocol used gives very acceptable results in the treatment of traumatic haemothorax.


Temporomandibular juxtaarticular chondroma: case report.
Chondromas are benign tumours composed of mature hyaline cartilage. We present here the first case in the English language medical literature of juxtaarticular chondroma of the temporomandibular joint in the parotid region. Within the rarity of cartilage disorders of the temporo-mandibular joint (TMJ), this particular condition is a diagnostic curiosity. The patient, a 54 year old woman, presented a right preauricular tumour of 3.5 cm. which had been developing for 4 years. It was not painful but there was a recent symptomology of TMJ dysfunction, with pain and clicks. The diagnostic possibilities of a parotid pleomorphic adenoma and of a cartilage tumour of the TMJ suggested a difficult preoperative differential diagnosis, which influenced our approach regarding therapy. The tumour was excised, preserving the parotid gland. This enabled us to confirm the histological diagnosis of chondroma, composed solely of chondroide tissue. We have described the clinical characteristics of our case, and carried out a review of the relevant literature, emphasising the differential diagnoses.


Tendon and pulleys at the metacarpophalangeal joint of a finger.
In fresh frozen traumatically amputated forearms with a constant tension of one kilogram on the flexor profundus tendon and the interphalangeal joints fixed in full extension by a Kirschner wire, the excursion of the tendon at the metacarpophalangeal joint and the force at the finger tip were correlated with different angles of flexion of the joint, first with the finger intact and then after varying amounts of advancement of the metacarpophalangeal joint pulley system. Pulley advancement increased the tendon excursion required to flex this joint and thus the mechanical advantage at this joint, but only when the joint was partly flexed. The extra excursion required at the metacarpophalangeal joint would be expected to weaken the interphalangeal joints at full flexion. Advancement also permitted ulnar-radial displacement of the tendon at the level of the metacarpophalangeal joint and hence could accentuate ulnar or radial drift. Pulley advancement is not recommended.


Loss of growth hormone responsiveness to normal physiological stimuli during pregnancy.
The response of pituitary GH to acute hyperglycaemia induced by 75 g oral glucose load in 73 pregnant women at various stages of gestation was examined. According to the age of gestation, patients were grouped into three groups: less than 20 weeks, between 20-30 and more than 30 weeks. Plasma glucose, GH and C-peptide were measured at fasting and then at 30, 60, 120 and 180 min following the glucose load. There was a significant increase in plasma GH concentration with weeks of gestation. The results also showed a loss of the normal physiological suppressive effects of hyperglycaemia on GH secretion in tests performed after 20 weeks of gestation. Only in tests performed before 20 weeks, there was a significant negative correlation between plasma glucose and GH values. These findings are consistent with recent reports suggesting an almost complete suppression of pituitary GH secretion by a placental variant of GH. These changes in the dynamics of GH secretion suggest that, in pregnancy, GH plays a more significant role than was previously thought.


Dendritic Cells Are Dispensable for T Cell Priming and Control of Acute Lymphocytic Choriomeningitis Virus Infection.
Dendritic cells (DCs) are considered to be the major APCs with potent activity for priming of naive CD4 and CD8 T cells. However, T cell priming can also be achieved by other APCs including macrophages, B cells, or even nonhematopoietic cell types. Systemic low-dose infection of mice with lymphocytic choriomeningitis virus (LCMV) results in massive expansion of virus-specific CD4 and CD8 T cells. To determine the role of DCs as APCs and source of type I IFNs in this infection model, we used ΔDC mice in which DCs are constitutively ablated because of expression of the diphtheria toxin α subunit within developing DCs. ΔDC mice showed lower serum concentrations of IFN-β and IL-12p40, but normal IFN-α levels during the first days postinfection. No differences were found for proliferation of transferred TCR-transgenic cells during the early phase of infection, suggesting that T cell priming occurred with the same efficiency in wild-type and ΔDC mice. Expansion and cytokine expression of endogenous LCMV-specific T cells was comparable between wild-type and ΔDC mice during primary infection and upon rechallenge of memory mice. In both strains of infected mice the viral load was reduced below the limit of detection with the same kinetic. Further, germinal center formation and LCMV-specific Ab responses were not impaired in ΔDC mice. This indicates that DCs are dispensable as APCs for protective immunity against LCMV infection.


Rapid diagnostic tests for malaria diagnosis in Cameroon: impact of histidine rich protein 2/3 deletions and lactate dehydrogenase gene polymorphism.
Malaria rapid diagnostic tests (mRDT) play a vital role in malaria control in endemic areas. In this study, histidine-rich protein (hrp) and lactate dehydrogenase (ldh) genes were genotyped in Plasmodium falciparum (Pf) and Plasmodium ovale (Po) spp. isolates. Deletions in P. falciparum hrp2/3 (pfhrp2/3) proteins and single nucleotide polymorphisms (SNPs) were analyzed. Twenty-four samples were analyzed for pfhrp2/3 gene deletions and 25 for SNPs in ldh gene (18 Pf and 7 Po spp.). Deletions in pfhrp2/3 genes were observed in 1.9% malaria positive isolates. The pfldh gene sequences showed one SNP at codon 272 (D272N) in 22.2% of samples while in Po spp., sequences were 100% similar to P. ovale curtisi but when compared to P. ovale wallikeri reference sequence, SNPs at positions 143 (P143S), 168 (K168N), 204 (V204I) were found. Findings suggest low prevalence in pfhrp2/3 genes and highlight the circulation of P. ovale curtisi in the studies areas.


Two-Electron Reduction versus One-Electron Oxidation of the Type 3 Pair in the Multicopper Oxidases.
Multicopper oxidases (MCOs) utilize an electron shuttling Type 1 Cu (T1) site in conjunction with a mononuclear Type 2 (T2) and a binuclear Type 3 (T3) site, arranged in a trinuclear copper cluster (TNC), to reduce O2 to H2O. Reduction of O2 occurs with limited overpotential indicating that all the coppers in the active site can be reduced via high-potential electron donors. Two forms of the resting enzyme have been observed in MCOs: the alternative resting form (AR), where only one of the three TNC Cu's is oxidized, and the resting oxidized form (RO), where all three TNC Cu's are oxidized. In contrast to the AR form, we show that in the RO form of a high-potential MCO, the binuclear T3 Cu(II) site can be reduced via the 700 mV T1 Cu. Systematic spectroscopic evaluation reveals that this proceeds by a two-electron process, where delivery of the first electron, forming a high energy, metastable half reduced T3 state, is followed by the rapid delivery of a second energetically favorable electron to fully reduce the T3 site. Alternatively, when this fully reduced binuclear T3 site is oxidized via the T1 Cu, a different thermodynamically favored half oxidized T3 form, i.e., the AR site, is generated. This behavior is evaluated by DFT calculations, which reveal that the protein backbone plays a significant role in controlling the environment of the active site coppers. This allows for the formation of the metastable, half reduced state and thus the complete reductive activation of the enzyme for catalysis.


Organophosphate pesticide exposure, hormone levels, and interaction with PON1 polymorphisms in male adolescents.
To examine the association between urinary metabolites of organophosphate (OP) pesticides and serum concentrations of thyroid and reproductive hormones in male adolescents and to assess the potential effect of interactions between OP pesticides and paraoxonase 1 (PON1) polymorphisms on hormone levels. Study subjects (N = 117) were male 16- to 17-year-olds from the Environment and Childhood (INMA)-Granada cohort in Spain. Concentrations of 3,5,6-trichloro-2-pyridinol (TCPy), a metabolite of chlorpyrifos/chlorpyrifos-methyl, 2-isopropyl-6-methyl-4-pyrimidinol (IMPy), a metabolite of diazinon, and diethylthiophosphate (DETP) and diethyldithiophosphate (DEDTP), non-specific metabolites of OP pesticides, were measured in a spot urine sample from each subject and adjusted for creatinine. Levels of reproductive hormones (total testosterone [TT], estradiol [E2], dehydroepiandrosterone sulfate [DHEAS], sex hormone binding globulin [SHBG], luteinizing hormone [LH], follicle stimulating hormone [FSH], anti-Müllerian hormone [AMH], insulin growth factor 1 [IGF-1], and prolactin), thyroid hormones (free thyroxine [FT4], total triiodothyronine [TT3], and thyroid stimulating hormone [TSH]), and PON1 Q192R and L55M polymorphisms were determined in blood drawn during the same clinical visit. Multiple linear regression models showed that detectable levels of TCPy were associated with an increase in DHEAS and decreases in E2, FSH, and AMH; detectable IMPy with increases in E2, DHEAS, FSH, AMH, and prolactin and decreases in SHBG and LH; and detectable DETP with marginally-significant increases in TT and TT3 and decreases in FSH, AMH, and prolactin. The effect of IMPy and DETP on DHEAS and TT levels, respectively, was higher in subjects that carried the PON1 55MM genotype, while the effect of TCPy, IMPy, and DETP on thyroid hormone levels was higher in PON1 192QR/RR or 55MM genotype carriers. In male adolescents, non-occupational exposure to OP pesticides was associated with several changes in reproductive and thyroid hormone levels, and the magnitude of some associations was greater in adolescents genetically more susceptible to OP pesticide exposure who carry the PON1 55MM genotype.