Simultaneous detection of multiple bone-related mRNAs and protein expression during osteoblast differentiation: polymerase chain reaction and immunocytochemical studies at the single cell level.
Messenger RNA expression analyzed by in situ hybridization and Northern analysis and protein expression analyzed biochemically or immunocytochemically have been used to study the developmental expression of various osteoblast (OB)-associated molecules. These approaches have shown that over a time course of OB differentiation in vivo and in vitro, the expression of macromolecules associated with OB cells changes. However, ambiguities in data from different approaches and in populations representative of cells at different developmental stages are extant. To begin to discriminate differentiation stages with more precision and to address intercellular heterogeneity, fetal rat calvaria cells were grown at low densities under conditions in which bone nodules form and mineralize and colonies were classified morphologically as fibroblastic or osteoblastic (early, intermediate, or mature). Whole discrete colonies and single cells from individual colonies were analyzed molecularly by a random amplification poly(A)-polymerase chain reaction (PCR) and for protein expression by immunocytochemistry; we analyzed the expression of known bone-related macromolecules (collagen type I, alkaline phosphatase, osteopontin, bone sialoprotein, and osteocalcin). Both PCR and immunocytochemistry revealed that different colony types were reproducibly distinguishable in their expression of either general (collagen type I) or bone-associated (alkaline phosphatase, osteopontin, bone sialoprotein, and osteocalcin) macromolecules, such that fibroblastic colonies were distinguishable from osteoblastic colonies and the latter could be subdivided into less mature or more mature osteoblastic colonies. While some aspects of the temporal differentiation sequence defined earlier were confirmed, several additional features were evident from these single cell-single colony studies. First, different repertoires of OB-associated markers were expressed in different cells, suggesting variation in the switch-on of the OB differentiation program and heterogeneity in the OB phenotype. Second, among colonies classified as fibroblastic on the basis of morphology heterogeneity was also evident and there were some cells expressing features consistent with their being osteoprogenitor cells. Our data support the hypothesis that individual fibroblastic and osteoblastic cells are heterogeneous in expression of marker molecules. We also conclude that individual cells and colonies analyzed by poly(A)-PCR will be useful in lieu of mass populations to extend investigation of stages in the progression of OB differentiation.


Circulating Anti-cytolethal Distending Toxin B and Anti-vinculin Antibodies as Biomarkers in Community and Healthcare Populations With Functional Dyspepsia and Irritable Bowel Syndrome.
Anti-cytolethal distending toxin B (CdtB) and anti-vinculin antibodies have been proposed as biomarkers that discriminate irritable bowel syndrome (IBS) diarrhea from inflammatory bowel disease; however, it is unknown whether they can also discriminate patients with IBS and IBS subtypes and functional dyspepsia (FD) from healthy individuals in the general population. We aimed to determine whether anti-CdtB and anti-vinculin can discriminate IBS and FD from health and from organic gastrointestinal (GI) disease. Adults were enrolled from 2 Australian studies: (i) a random, population-based study (n = 331) with subjects diagnosed with IBS (n = 63) or FD (n = 61) by modified Rome III criteria or healthy control subjects (n = 246) who did not meet criteria for IBS and/or FD and (ii) an outpatient-based study with subjects diagnosed with IBS (n = 256) and/or FD (n = 55) or organic GI disease (n = 182) by an independent clinician. Serum levels of anti-CdtB/anti-vinculin antibodies were determined by enzyme-linked immunosorbent assay. There was a significantly higher mean value of anti-CdtB in FD vs healthy controls (mean = 2.46 [SD = 0.72] vs mean = 2.14 [SD = 0.77]; P = 0.005) and IBS/FD overlap vs healthy controls (mean = 2.47 [SD = 0.78] vs mean = 2.14 [SD = 0.77]; P = 0.02). There were no significant differences in anti-CdtB in IBS and FD outpatients or IBS/FD subgroups compared with patients with organic GI disease. In terms of anti-vinculin, there were no significant differences between IBS and FD and healthy controls or between IBS and FD and organic GI disease controls. We did not confirm that anti-CdtB/anti-vinculin discriminated IBS diarrhea from organic GI disease in Australian subjects. However, we did find higher anti-CdtB in FD and IBS/FD overlap vs healthy controls. Postinfectious FD may be more common than currently recognized.


Immunolocation of TNF-alpha/cachectin in human melanoma cells: studies on co-cultivated malignant melanoma.
We have investigated the ability of metastatic cells to produce the macrophage cytokine, TNF-alpha/cachectin, as these cells have macrophage-like properties such as infiltration and migration. We looked for TNF-alpha/cachectin in three tumor cell lines derived from human malignant melanomas and six co-cultivated malignant melanomas derived, in vitro, from these three cell lines plus angioma fibroblasts. Immunohistochemistry with an anti-TNF-alpha/cachectin monoclonal antibody showed that TNF-alpha/cachectin was produced by two of the three parent melanoma cell lines. All the tumor cells in both the co-cultivated malignant melanomas and their in vitro tumorous nodules produced TNF-alpha/cachectin, even those derived from the melanoma cell line, which originally did not. The results clearly show that TNF-alpha/cachectin can be produced by non-hematopoietic tumor cells. A co-cultivated tumor model prepared from other types of human tumor cell lines promises to provide a useful tool for exploring the relationship between TNF-alpha/cachectin and oncogenesis.


Poisoned Wine: Regulation, Chemical Analyses, and Spanish-French Trade in the 1930s.
This paper describes the resources, scientific spaces, and experts involved in the study of a mass poisoning caused by the drinking of arsenic-contaminated wine exported from Spain to France in 1932. Local and international periodicals record the poisoning of 300 French sailors, and stressed the commercial implications of the case. We discuss the reports prepared by different experts (mainly physicians, agricultural engineers, and customs chemists). Their work was not limited to preparing technical publications or chemical analyses; they also actively defended the quality of their local wine, and played a major role in the discussions regarding the regulation of the international wine market in the 1930s, when new standards regarding the analysis of wine were being considered. Curiously, this well-publicised case of mass poisoning did not have any noticeable consequences in the international regulation of wine. This absence of subsequent regulatory action and the role of experts are central topics of the paper.


The predictability of infant visual-evoked response testing on future visual acuity.
We reviewed the records of 27 infants with abnormal eye examinations and visual-evoked response (VER) testing (mean age, 10.5 months) who subsequently underwent a long-term follow-up ophthalmology examination (mean duration, 41 months). The infants were initially diagnosed with various ocular disorders including cortical blindness (eight), optic nerve hypoplasia (six), congenital cataract (two), and retinopathy of prematurity (one). Standard optotype visual-acuity determinations were available in the follow-up records of 11 children (21 eyes), and fixation behavior was obtained in the remaining 16 children (32 eyes). Results revealed that pattern-reversal VER P1 latency was predictive (87%) of whether visual acuity was equal to (or better than) or worse than 20/100 and whether a patient would have good fixation behavior (fix and follow, FF) or poor fixation (no FF) (86%) (P less than .001). Although flash VER P1 latency was also predictive of later visual acuity or good fixation (73%), it was not statistically significant. Pattern VER P1 amplitude and flash VER P1 amplitude were not predictive of later visual function. The predictive power of pattern VER P1 latency for later visual function probably relates to its reflection of macular function and low variability. An analysis of the variability of each of the four VER factors in normal infants (n = 50) indicated that pattern VER P1 latency was the least variable, and consequently most sensitive, VER factor for detecting and quantifying pathology. Overall, the results of this retrospective study suggest that pattern VER P1 latency may have important predictive power for later visual function in infants with an initially abnormal ophthalmologic examination.


The evolution of rod photoreceptors.
Photoreceptors in animals are generally of two kinds: the ciliary or c-type and the rhabdomeric or r-type. Although ciliary photoreceptors are found in many phyla, vertebrates seem to be unique in having two distinct kinds which together span the entire range of vision, from single photons to bright light. We ask why the principal photoreceptors of vertebrates are ciliary and not rhabdomeric, and how rods evolved from less sensitive cone-like photoreceptors to produce our duplex retina. We suggest that the principal advantage of vertebrate ciliary receptors is that they use less ATP than rhabdomeric photoreceptors. This difference may have provided sufficient selection pressure for the development of a completely ciliary eye. Although many of the details of rod evolution are still uncertain, present evidence indicates that (i) rods evolved very early before the split between the jawed and jawless vertebrates, (ii) outer-segment discs make no contribution to rod sensitivity but may have evolved to increase the efficiency of protein renewal, and (iii) evolution of the rod was incremental and multifaceted, produced by the formation of several novel protein isoforms and by changes in protein expression, with no one alteration having more than a few-fold effect on transduction activation or inactivation.This article is part of the themed issue 'Vision in dim light'.


Solvolysis of 4-methylcyclohexylidenemethyliodonium salt: chirality probe approach to a primary vinyl cation intermediate.
Optically active 4-methylcyclohexylidenemethyl(aryl)iodonium tetrafluoroborate (1.BF(4)(-)) was prepared and its solvolysis was carried out at 60 degrees C in various solvents. The main product is optically active 4-methylcycloheptanone (or its enol derivative) in unbuffered solvents, accompanied by the iodoarene. The rearranged product always maintains the optical purity of the starting 1. Its stereochemistry conforms to a mechanism involving the rearrangement via the sigma-bond participation in departure of the nucleofuge, followed by trapping of the resulting chiral 5-methylcyclohept-1-enyl cation with a nucleophilic solvent. That is, the achiral, primary vinyl cation is not involved during the reaction. The unrearranged substitution product is also obtained in a minor fraction in unbuffered methanol, ethanol, and acetic acid, but not in trifluoroethanol or hexafluoro-2-propanol: the methoxy product from methanolysis is largely racemized, but the acetolysis product is obtained mainly via retention of configuration. Reactions of 1 with bromide, acetate, and trifluoroacetate in chloroform give unrearranged substitution products in different degrees of inversion. These unrearranged products are concluded to be formed via the direct nucleophilic substitutions. Added bases such as sodium acetate in methanol lead to the unrearranged methoxy products of complete racemization, which is ascribed to the alpha elimination (to give an alkylidenecarbene) followed by the solvent insertion.


The prognostic and therapeutic implications of cytologic atypia in patients with endometrial hyperplasia.
From 1970 to 1992, 136 patients with a histologic diagnosis of endometrial hyperplasia underwent total abdominal hysterectomy at the University of Kentucky Medical Center. Slides of the curettage or biopsy specimens were reviewed and classified according to the International Society of Gynecologic Pathologists System as simple or complex endometrial hyperplasia with or without cytologic atypia. Slides of the hysterectomy specimens were likewise reviewed independently and classified according to the same system. Eighty-two patients had a preoperative diagnosis of simple or complex endometrial hyperplasia without atypia. There were no cases of occult endometrial carcinoma in the hysterectomy specimens of these patients. Simple or complex hyperplasia with atypia was present in 54 patients and endometrial adenocarcinoma was observed in 19 of these cases (35%). The International Federation of Gynecology and Obstetrics stage and histologic grade of these patients was as follows: Stage IA grade 1--5; Stage IB grade 1--10; Stage IB grade 2--1; Stage IC grade 1--1; Stage IC grade 2--1; and Stage IIIA grade 2--1. The risk of associated endometrial cancer in patients with atypical hyperplasia was independent of age, diabetes mellitus, hypertension, or the use of exogenous estrogens. All patients with endometrial cancer have been followed for 1-12 years (mean 3.0 years) after therapy and no patient has experienced tumor recurrence. These data suggest that there is a significant risk of endometrial cancer in patients with histologic evidence of atypical endometrial hyperplasia on curettage or biopsy. At the time of surgery, patients with atypical endometrial hyperplasia should have careful inspection of the uterine specimen. Any endometrial tissue suspicious for malignancy should be examined histologically, and if cancer is confirmed, complete surgical staging should be performed.


Performance of host-response biomarkers to risk-stratify children with pneumonia in Bhutan.
Pneumonia is the leading cause of post-neonatal death amongst children under five years of age; however, there is no simple triage tool to identify children at risk of progressing to severe and fatal disease. Such a tool could assist for early referral and prioritization of care to improve outcomes and enhance allocation of scarce resources. We compared the performance of inflammatory and endothelial activation markers in addition to clinical signs or scoring scales to risk-stratify children hospitalized with pneumonia at the national referral hospital of Bhutan with the goal of predicting clinical outcome. Of 118 children, 31 evolved to a poor prognosis, defined as either mortality, admission in the paediatric intensive care unit, requirement of chest drainage or requirement of more than five days of oxygen therapy. Soluble triggering receptor expressed on myeloid cells 1 (sTREM-1) was the best performing biomarker and performed better than clinical parameters. sTREM-1 levels upon admission had good predictive accuracy to identify children with pneumonia at risk of poor prognosis. Our findings confirm that immune and endothelial activation markers could be proactively used at first encounter as risk-stratification and clinical decision-making tools in children with pneumonia; however, further external validation is needed.


Release of enzymes from human leucocytes during incubation with Neisseria gonorrhoeae.
The effect of Neisseria gonorrhoeae on release of enzymes from human leucocytes was determined. Supernatants from incubation mixtures containing leucocytes and gonococci were assayed for activity of the cytoplasmic enzyme, lactic acid dehydrogenase, as well as for activity of the hydrolytic enzymes, β-glucuronidase and lysozyme, which are found primarily in leucocyte granules. Thirty-minute incubation of leucocytes with pilated T1 gonococci resulted in a negligible release of lactic acid dehydrogenase and little release of β-glucuronidase even at bacteria to leucocyte ratios as high as 50 to 1. Lysozyme release, however, was significant at this ratio and at 20 to 1 but not at 5 to 1. Incubation with non-pilated T4 bacteria yielded no significant release of lactic acid dehydrogenase or β-glucuronidase, but it caused a significant release of lysozyme at bacteria to leucocyte ratios as low as 2 to 1. These results suggested that the lysozyme release might be related to the degree of phagocytic activity since, at low ratios, T4 was readily ingested but T1 was not. Consistent with this hypothesis, serum which promoted the phagocytosis of the pilated gonococci also stimulated lysozyme release at low ratios of T1 to leucocyte. Absorption of the serum with T1 abolished the opsonic effect and markedly diminished the amount of lysozyme released.


Patients over 70 years of age with moderate ischemic mitral regurgitation undergoing surgical revascularization plus mitral valve repair: insights from a single-center study of propensity-matched data.
Elderly patients, compared with the young, have a higher burden of surgical risk factors with reduced functional capacity and increased comorbidities conditions, and may have worse clinical outcomes. So far, few reports have focused on clinical outcomes of patients over 70 years of age with moderate chronic ischemic mitral regurgitation (IMR) undergoing mitral valve repair at the time of coronary artery bypass grafting (CABG). This single-center study of propensity-matched data attempts to answer a question: compared with patients with age of 70 or less, whether patients over 70 years of age with moderate IMR undergoing CABG plus mitral valve repair receive poor outcomes. All eligible patients were included in this study and were entered into either an elderly group (n=142) or a control group (n=182) according to patients' age. In-hospital outcomes (consisting of surgical mortality and major postoperative morbidity) and midterm clinical outcomes (including all-cause mortality and recurrent mitral regurgitation) were compared after propensity score matching (1:1). Using propensity-score matching, 103 pairs of patients were successfully established in a 1:1 ratio. No significant differences between the two matched groups were found with regard to surgical mortality (5.8% vs. 3.9%, P=0.754) and major postoperative morbidity. A total of 184 patients (91 in the elderly group and 93 in the control group) received regular follow-up visit with the median duration of 38 months [interquartile range (IQR), 27-56 months]. There were not any significant differences between the two matched groups regarding overall survival and recurrent IMR-free survival (stratified log-rank P=0.185 and stratified log-rank P=0.453, respectively). The elderly group as compared to the control group did not affect midterm mortality via cox proportional hazard regression (propensity score adjusted hazard ratio, 1.143; 95% confidence interval, 0.761-1.943; P=0.285). Patients over 70 years of age with moderate chronic IMR undergoing combined CABG and mitral valve repair may receive favorable in-hospital and midterm clinical outcomes.


Labor Curves in Multiparous Women Related to Interbirth Intervals.
Previous studies have shown that risk of cesarean section increases among multiparous women as interbirth interval increases. One possibility is that progress of labor may vary with interbirth interval, such that with longer intervals, labor curves of multiparas more closely resemble those of nulliparas. We sought to define labor curves among a cohort of multiparas with varying interbirth intervals. This was a retrospective cohort study of term multiparas with known interval from last delivery and only vaginal deliveries. Subjects were grouped by interval between the studied pregnancy and the most recent birth: 0 to 59, 60 to 119, and ≥120 months. Statistical analysis was performed using linear mixed effects model. Group slopes and intercepts were compared using model t-tests for individual effects. Length of second stage was compared using a Wilcoxon's rank-sum test. Groups did not differ significantly in demographic or obstetrical characteristics. Rate of dilation was similar between the 0 to 59 and 60 to 119 month groups (p = 0.38), but faster in the ≥120 month group compared with the 60 to 119 month group (p = 0.037). Median duration of second stage increased slightly with increased interbirth interval (p = 0.003). Prolonged interbirth interval is not associated with slower active phase of labor.


Low oxygenation index and pulmonary artery hypertension in predicting early death from adult respiratory distress syndrome (ARDS).
Hemodynamic and respiratory parameters were continuously monitored in 45 septic shock patients, 15 of whom developed adult respiratory distress syndrome (ARDS). Low oxygenation index (OI = PaO2/FIO2), pulmonary artery hypertension (high mean pulmonary artery pressure, MPAP) and elevated pulmonary vascular resistance (PVR) were observed in all ARDS, as well as in non-ARDS septic patients, as a baseline. These same pulmonary factors were compared between those who survived and those who died during the first few days (early fatalities) in both the ARDS group (5 patients) and the non-ARDS group (8 patients). The results showed that the early fatalities in the ARDS group had a significantly lower (p less than 0.001) OI than the survivors, as well as a lower MPAP (p less than 0.01). In the non-ARDS group, the MPAP of the early fatalities was significantly lower (p less than 0.01) than that of survivors, but their OI was not significantly lower. PVR, when compared between groups (ARDS versus non-ARDS) or between subgroups within each group, was elevated concurrently with the elevation of MPAP. It is concluded that patients with septic shock and ARDS who show a severely depressed OI and a modestly elevated MPAP and PVR during the first few days can be predicted to have a poor outcome.


Dissociation between PGC-1alpha and GLUT-4 expression in skeletal muscle of rats fed a high-fat diet.
It has recently been reported that a 4-wk high-fat diet gradually increases skeletal muscle peroxisome proliferator activated receptor (PPAR) gamma coactivator-1alpha (PGC-1alpha) protein content, which has been suggested to regulate GLUT-4 gene transcription. However, it has not been reported that a high-fat diet enhances GLUT-4 mRNA expression and protein content in skeletal muscle, suggesting that an increase in PGC-1alpha protein content is not sufficient to induce muscle GLUT-4 biogenesis in a high-fat fed animal. Therefore, we first evaluated the relationship between PGC-1alpha and GLUT-4 expression in skeletal muscle of rats fed a high-fat diet for 4 wk. The PGC-1alpha protein content in rat epitrochlearis muscle significantly increased by twofold after the 4-wk high-fat diet feeding. However, the high-fat diet had no effect on GLUT-4 protein content and induced a 30% decrease in GLUT-4 mRNA expression in rat skeletal muscle (p<0.05). To clarify the mechanism by which a high-fat diet downregulates GLUT-4 mRNA expression, we next examined the effect of PPARdelta activation, which is known to occur in response to a high-fat diet, on GLUT-4 mRNA expression in L6 myotubes. Incubation with 500 nM GW501516 (PPARdelta activator) for 24 h significantly decreased GLUT-4 mRNA in L6 myotubes. Taken together, these findings suggest that a high-fat diet downregulates GLUT-4 mRNA, possibly through the activation of PPARdelta, despite an increase in PGC-1alpha protein content in rat skeletal muscle, and that a posttranscriptional regulatory mechanism maintains GLUT-4 protein content in skeletal muscle of rats fed a high-fat diet.


Statin Use and Delirium Risk: An Updated Systematic Review and Meta-Analysis.
Findings on the association of statin use with delirium risk are inconsistent. Is statin use associated with delirium risk? We searched PubMed, the Cochrane Library, and the EMBASE database, limiting the search to human patients and articles in English published until December 31, 2021. The effect size and 95% confidence interval (CI) were defined as the odds ratio (OR) and 95% CI, respectively, to indicate the difference in the incidence of delirium between statin use and nonuse groups. A random-effects model was selected in the case of high heterogeneity of study populations. We used funnel plots, Egger test, Duval and Tweedie trim-and-fill approach, and the classic fail-safe N to assess publication bias. Of a total of 264 identified studies, 13 were selected for the qualitative review-4 RCTs and 9 observational cohort studies. Statin use was not associated with low delirium risk (pooled OR, 0·82; 95% CI, 0·64-1·04; P = 0·09). Substantial statistical heterogeneity was observed ( I2 , 90%). Visual inspection of the funnel plot of ORs from the studies revealed symmetry. Using the Grading of Recommendations Assessment, Development, and Evaluation approach, we assigned the evidence a rating of C and a weak recommendation for this review. Statin use is not associated with delirium risk. More comprehensive RCTs are required to confirm the results.


Application of risk prediction models to lung cancer screening: a review.
Globally, lung cancer is the leading cause of cancer death and is a major public health problem. Because lung cancer is usually diagnosed at an advanced stage, survival is generally poor. In recent decades, clinical advances have not led to marked improvements in outcomes. A recent advance of importance arose when the National Lung Screening Trial (NLST) findings indicated that low-dose computed tomography screening of high-risk individuals can lead to a lung cancer mortality reduction of 20%. NLST identified high-risk individuals using the following criteria: age 55 to 74 years; ≥30 pack-years of smoking; and number of years since smoking cessation ≤15 years. Medical screening is most effective when applied to high-risk individuals. The NLST criteria for high risk were practical for enrolling individuals into a clinical trial but are not optimal for risk estimation. Lung cancer risk prediction models are expected to be superior. Indeed, recently, 3 studies have provided quantitative evidence that selection of individuals for lung screening on the basis of estimates from high-quality risk prediction models is superior to using NLST criteria or similar criteria, such as the United States Preventive Services Task Force (USPSTF) criteria. Compared with NLST/USPSTF criteria, selection of individuals for screening using high-quality risk models should lead to fewer individuals being screened, more cancers being detected, and fewer false positives. More lives will be saved with greater cost-effectiveness. In this paper, we review methodological background for prediction modeling, existing lung cancer risk prediction models and some of their findings, and current issues in lung cancer risk prediction modeling and discuss future research.


Bayesian shrinkage analysis of quantitative trait Loci for dynamic traits.
Many quantitative traits are measured repeatedly during the life of an organism. Such traits are called dynamic traits. The pattern of the changes of a dynamic trait is called the growth trajectory. Studying the growth trajectory may enhance our understanding of the genetic architecture of the growth trajectory. Recently, we developed an interval-mapping procedure to map QTL for dynamic traits under the maximum-likelihood framework. We fit the growth trajectory by Legendre polynomials. The method intended to map one QTL at a time and the entire QTL analysis involved scanning the entire genome by fitting multiple single-QTL models. In this study, we propose a Bayesian shrinkage analysis for estimating and mapping multiple QTL in a single model. The method is a combination between the shrinkage mapping for individual quantitative traits and the Legendre polynomial analysis for dynamic traits. The multiple-QTL model is implemented in two ways: (1) a fixed-interval approach where a QTL is placed in each marker interval and (2) a moving-interval approach where the position of a QTL can be searched in a range that covers many marker intervals. Simulation study shows that the Bayesian shrinkage method generates much better signals for QTL than the interval-mapping approach. We propose several alternative methods to present the results of the Bayesian shrinkage analysis. In particular, we found that the Wald test-statistic profile can serve as a mechanism to test the significance of a putative QTL.


Primary repair with semitendinosus tendon augmentation of acute anterior cruciate ligament injuries.
A retrospective review of 72 acute ACL injuries in 70 athletically active patients (50 recreational and 20 competitive athletes) treated with primary repair and semitendinosus tendon augmentation was conducted (mean follow-up time, 38.5 months). All patients had open primary multiple loop depth-varying suture repair and semitendinosus tendon augmentation at a mean injury to surgery interval of 9.1 days. Fifty-one cases (70.8%) were supplemented with an extraarticular procedure consisting in all cases of an iliotibial band lateral sling reinforcement. All patients underwent 6 weeks of postoperative immobilization followed by a graduated rehabilitation regimen lasting a mean 7.2 months. In 22 of the ACL tears, no other associated injury could be defined at arthrotomy, while in 27, significant medial collateral ligament injuries were noted. Lateral meniscal injuries (24) were more commonly noted than medial meniscal injuries (19). Good to excellent subjective results were reported in 82%, while 77.1% returned to preinjury sport level and participation without limitation. One patient developed "giving way" symptoms and overall, only four patients did not return to sports participation. Objective examination revealed 93.1% to have a 1+ or less Lachman test and 86.1% to have a negative pivot shift. Of 60 knees tested, 93.3% had KT-1000 side-to-side difference values (measured at 25 degrees +/- 5 degrees of flexion with an 89 N load) of less than or equal to 3 mm of anterior displacement. A 100 point Hospital for Special Surgery ligament rating score was used with 25 points assigned to subjective, 45 points to objective, and 30 points to functional assessment. The mean score at followup was 93.1. Analysis of results in patients with or without an extraarticular reinforcement revealed no difference in objective outcome. The incidence of patellofemoral pain was 27.8%. Thirty knees had some degree of loss of range of motion. Loss of terminal flexion was noted in 29, while 5 had loss of terminal extension. No correlation could be found between patellofemoral pain and diminished range of motion. Generalized ligamentous laxity was found in 37.1% of the patients; this was not associated with a poor result. This study suggests that primary repair with semitendinosus tendon augmentation of acute ACL injuries with a graduated rehabilitation regimen provides good subjective, and excellent functional and objective, results in active patients that were followed for more than 3 years.


Transplantation of liver organoids in the omentum and kidney.
Liver organoids were reconstructed by mouse-immortalized hepatocytes and nonparenchymal cells (sinusoidal endothelial cells and hepatic stellate cells) in a radial-flow bioreactor (RFB). A biodegradable apatite-fiber scaffold (AFS) was used as a scaffold packed in the RFB, which enables three-dimensional cell cultures. The organoids cocultured in the RFB showed a liver-like structure with high-density layers of hepatocytes and the formation of vessel-like structures. A liver organoid consisting of three cocultured cells was transplanted under the kidney capsule (kidney group) or into the omentum (omentum group) using BALB/c nude mice. Transplanted liver organoids survived in the kidney or omentum. The expression of mRNAs of albumin, connexin 26 and 32, hepatocyte nuclear factor 4α, and glucose-6-phosphatase was increased in both groups at 8 weeks after transplantation in comparison to the pretransplant status. Tyrosine aminotransferase appeared only in the omentum group. The results suggested that the functions of liver organoids differed depending on the transplanted site in the recipient animals.


Medication Administration Practices in United States' Schools: A Systematic Review and Meta-synthesis.
Schools often provide medication management to children at school, yet, most U.S. schools lack a full-time, licensed nurse. Schools rely heavily on unlicensed assistive personnel (UAP) to perform such tasks. This systematic review examined medication management among K-12 school nurses. Keyword searches in three databases were performed. We included studies that examined: (a) K-12 charter, private/parochial, or public schools, (b) UAPs and licensed nurses, (c) policies and practices for medication management, or (d) nurse delegation laws. Three concepts were synthesized: (a) level of training, (b) nurse delegation, and (c) emergency medications. One-hundred twelve articles were screened. Of these, 37.5% (42/112) were comprehensively reviewed. Eighty-one percent discussed level of training, 69% nurse delegation, and 57% emergency medications. Succinct and consistent policies within and across the United States aimed at increasing access to emergency medications in schools remain necessary.


The Protein Extract of Chlorella minutissima Inhibits The Expression of MMP-1, MMP-2 and MMP-9 in Cancer Cells through Upregulation of TIMP-3 and Down Regulation of c-Jun.
Considering the bioactivities exhibited by microalgae, the effect of protein extract of Chlorella minutissimma (CP extract) was investigated on the expression of human matrix metalloproteinases-1 (MMP-1) in the breast cancer cell line MDA-MB231, and that of MMP-2 and -9 in hepatocellular cancer cell line HepG2 at different expression levels. The study aimed identification and analysis of inhibitory activity of microalgal components extracted from Chlorella minutissima against human MMPs. In this experimental study, we analysed the effect of Chlorella extracts on MMP-1, -2, and -9 expression at various levels. Gelatin zymography was performed to study the inhibitory effect of Chlorella exracts on human gelatinases at the activity level, followed by western blotting to analyse the expression of all three MMPs at the protein level. The similar effect at the mRNA level along with the probable mechanism underlying inhibition of MMPs was assessed using real-time polymerase chain reaction (PCR). The results reveal that the treatment with CP extract decreased the mRNA expression of MMP-1, MMP-2, and MMP-9 by 0.26-, 0.29-, and 0.40-fold, respectively, at 20 μg/ml concentration as well as inhibited the activity of MMP-2 and MMP-9 by 37.56 and 42.64%, respectively, at 15 μg/ml concentration. Additionally, upregulated mRNA expression of tissue inhibitor of metalloproteinases-3 (TIMP-3) by 1.68-fold was seen in HepG2 cells at 20 μg/ml concentration treatment group. However, CP extract did not induce any change in the mRNA expression of the TIMP-1, -2 and -4 in HepG2 and TIMP-1, -2, -3 and -4 in MDA-MB231 cells. Activator protein-1 (AP-1)-dependent c-Jun-mediated transcriptional regulation of MMP-1, -2, and -9 was also studied to elucidate the appropriate mechanism involved in the inhibition of MMPs. The CP extract successfully inhibited MMP-1, -2, and -9 at different expression levels through TIMP-3 upregulation and c-Jun downregulation.


Validation of the YLS/CMI on an Australian Juvenile Offending Population.
There is a wealth of research that shows juvenile justice systems that utilize structured and validated assessment tools, such as the YLS/CMI, are far more effective at reducing rates of recidivism than those who do not. In line with this research, the Department of Justice (DoJ) in Western Australia adopted the YLS/CMI as the standard risk assessment tool for evaluating the criminogenic risk and needs of youth entering the justice system. While there is evidence supporting the utility of the YLS/CMI in predicting recidivism, there is little research demonstrating its effectiveness in Australian juvenile populations and no such research in a West Australian population. There is also a lack of research on the utility of the tool with young Indigenous offenders, which is particularly concerning given the significant overrepresentation of Indigenous people in the Australian criminal justice system. Our study was the first to examine the reliability and predictive validity of the YLS/CMI on a sample of West Australian juvenile offenders. In this paper, we present the results of two analyses. The first examines the properties of the YLS/CMI in a cohort of 4,653 juvenile offenders in Western Australia, including factor structure, internal consistency, and differences between male and female youth and between Indigenous and non-Indigenous youth. Consistent with our hypotheses, the tool demonstrated excellent internal consistency (α = .91) and cross-validation analyses identified significant differences between groups on total YLS scores and risk domains. The second analysis examines the predictive validity of the YLS/CMI in a subsample of 921 youth with a minimum follow-up period of 2 years. The overall recidivism rate was 74.8% and there were differences in scores and recidivism rates for Indigenous compared to non-Indigenous youth, although the predictive accuracies in terms of AUC were similar (c. = 0.65 and 0.66, respectively).


Midbrain hemorrhage presenting with isolated downward gaze palsy: A case report.
Diseases presenting with only downward gaze palsy are extremely rare particular in cerebral hemorrhage. A 63-year-old man with no medical history developed a downward-dominant vertical gaze paralysis with a convergence disorder. Computed tomography and magnetic resonance imaging showed a small hemorrhage of 13 mm in diameter in the right midbrain tegmental area. The patient was conservatively treated. His symptoms showed a gradual improvement from upward gaze paralysis and convergence disorder followed by downward gaze paralysis. All symptoms disappeared in 3 weeks after the onset. The center of vertical eye movement was thought to be the rostral interstitial nucleus of medial longitudinal fasciculus (riMLF) and posterior commissure (PC). In this case, bilateral riMLF and PC were impaired, resulting in bilateral vertical ocular motility disorder with upward gaze paralysis. Brainstem hemorrhage rarely presents with vertical gaze palsy. Most are cerebral infarctions, and few are due to cerebral hemorrhage. This case was important for better understanding the pathophysiology of an ocular motility disorder. We also summarized the characteristics of isolated vertical gaze palsy caused by cerebral hemorrhage.


A set-point model with oscillatory behavior predicts the time course of 8-OH-DPAT-induced hypothermia.
Agonists for the 5-hydroxytryptamine (HT)(1A) receptor induce a hypothermic response that is believed to occur by lowering of the body's set-point temperature. We have developed a physiological model that can be used to predict the complex time course of the hypothermic response after administration of 5-HT(1A) agonists to rats. In the model, 5-HT(1A) agonists exert their effect by changing heat loss through a control mechanism with a thermostat signal that is proportional to the difference between measured and set-point temperature. Agonists exert their effect in a direct concentration-dependent manner, with saturation occurring at higher concentrations. On the basis of simulations, it is shown that, depending on the concentration and the intrinsic efficacy of a 5-HT(1A) agonist, the model shows oscillatory behavior. The model was successfully applied to characterize the complex hypothermic response profiles after administration of the reference 5-HT(1A) agonists R-8-hydroxy-2-(di-n-propylamino)tetralin (R-8-OH-DPAT) and S-8-OH-DPAT. This analysis revealed that the observed difference in effect vs. time profile for these two reference agonists could be explained by a difference in in vivo intrinsic efficacy.


Members of the 30- to 32-kilodalton mycolyl transferase family (Ag85) from culture filtrate of Mycobacterium avium subsp. paratuberculosis are immunodominant Th1-type antigens recognized early upon infection in mice and cattle.
The characterization of protective antigens is essential for the development of an effective, subunit-based vaccine against paratuberculosis. Surface-exposed and secreted antigens, present abundantly in mycobacterial culture filtrate (CF), are among the well-known protective antigens of Mycobacterium tuberculosis and Mycobacterium bovis. Culture filtrate, prepared from Mycobacterium avium subsp. paratuberculosis ATCC 19698 grown as a surface pellicle on synthetic Sauton medium, was strongly and early recognized in experimentally infected B6 bg/bg beige mice and cattle, as indicated by elevated spleen cell gamma interferon (IFN-gamma) secretion and lymphoproliferative responses of peripheral blood mononuclear cells, respectively. Strong proliferative and ex vivo IFN-gamma responses against antigen 85 (Ag85) complex (a major protein component from M. bovis BCG culture filtrate) could be detected in cattle as early as 10 weeks after oral M. avium subsp. paratuberculosis infection. Synthetic peptides from the Ag85A and Ag85B components of this complex were strongly recognized, whereas T-cell responses were weaker against peptides from the Ag85C protein. A promiscuous T-cell epitope spanning amino acids 145 to 162 of Ag85B (identical sequence in M. bovis and M. avium subsp. paratuberculosis) was identified in experimentally infected cattle. Finally, young calves, born from cows with confirmed paratuberculosis, demonstrated proliferative responses to purified, recombinant Ag85A and Ag85B from M. avium subsp. paratuberculosis. These results indicate that the M. avium subsp. paratuberculosis Ag85 homologues are immunodominant T-cell antigens that are recognized early in experimental and natural infection of cattle.


How clean is "clean"? Regulations and standards for workplace clothing and personal protective equipment.
To compile current regulations and advisory recommendations on cleanliness of worker clothing and personal protective equipment and to evaluate the adequacy of criteria for determining whether cleanliness has been achieved. Systematic review of information provided by federal agencies (eg, OSHA, MSHA, and NIOSH), nongovernmental advisory bodies (eg, ACGIH, AIHA, and ANSI), and manufacturers of protective clothing and equipment. We identified an array of terms describing "cleanliness" and the processes for achieving "cleanliness" that were almost never defined in regulations and recommendations. We also found a general lack of criteria for determining whether cleanliness and/or sterility have been achieved. There is need to harmonize cleanliness-related terminology, establish best practices for equipment cleaning and sterilization, implement a signage systems to provide equipment-specific cleaning instructions, and adopt objective criteria for determining what is "clean."


Invisible barriers: anthropogenic impacts on inter- and intra-specific interactions as drivers of landscape-independent fragmentation.
Anthropogenically induced fragmentation constitutes a major threat to biodiversity. Presently, conservation research and actions focus predominantly on fragmentation caused directly by physical transformation of the landscape (e.g. deforestation, agriculture, urbanization, roads, etc.). While there is no doubt that landscape features play a key role in fragmenting populations or enhancing connectivity, fragmentation may also come about by processes other than the transformation of the landscape and which may not be readily visible. Such landscape-independent fragmentation (LIF) usually comes about when anthropogenic disturbance alters the inter- and intra-specific interactions among and within species. LIF and its drivers have received little attention in the scientific literature and in the management of wildlife populations. We discuss three major classes of LIF processes and their relevance for the conservation and management of species and habitats: (i) interspecific dispersal dependency, in which populations of species that rely on other species for transport and propagation become fragmented as the transporting species declines; (ii) interspecific avoidance induction, where species are excluded from habitats and corridors owing to interspecific interactions resulting from anthropogenically induced changes in community structure (e.g. exclusions by increased predation pressure); and (iii) intraspecific behavioural divergence, where populations become segregated owing to anthropogenically induced behavioural differentiation among them. This article is part of the theme issue 'Linking behaviour to dynamics of populations and communities: application of novel approaches in behavioural ecology to conservation'.


Simulation Studies Provide Evidence of Aerosol Transmission of SARS-CoV-2 in a Multi-Story Building via Air Supply, Exhaust and Sanitary Pipelines.
A cross-layer non-vertical transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) occurred in a quarantine hotel in Guangzhou, Guangdong Province, China in June 2021. To explore the cross-layer transmission path and influencing factors of viral aerosol, we set up different scenarios to carry out simulation experiments. The results showed that the air in the polluted room can enter the corridor by opening the door to take food and move out the garbage, then mix with the fresh air taken from the outside as part of the air supply of the central air conditioning system and re-enter into different rooms on the same floor leading to the same-layer transmission. In addition, flushing the toilet after defecation and urination will produce viral aerosol that pollutes rooms on different floors through the exhaust system and the vertical drainage pipe in the bathroom, resulting in cross-layer vertical transmission, also aggravating the transmission in different rooms on the same floor after mixing with the air of the room and entering the corridor to become part of the air supply, and meanwhile, continuing to increase the cross-layer transmission through the vertical drainage pipe. Therefore, the air conditioning and ventilation system of the quarantine hotel should be operated in full fresh air mode and close the return air; the exhaust volume of the bathroom should be greater than the fresh air volume. The exhaust pipe of the bathroom should be independently set and cannot be interconnected or connected in series. The riser of the sewage and drainage pipeline of the bathroom should maintain vertical to exhaust independently and cannot be arbitrarily changed to horizontal pipe assembly.


Comparison of porcine epidemic diarrhea viruses from Germany and the United States, 2014.
Since 2013, highly virulent porcine epidemic diarrhea virus has caused considerable economic losses in the United States. To determine the relation of US strains to those recently causing disease in Germany, we compared genomes and found that the strain from Germany is closely related to variants in the United States.


Microfluorometric measurements of cytoplasmic calcium in chief and oxyphil parathyroid cells of adenomatous and hyperplastic glands and of normal-sized glands associated with adenomas.
The effects of extracellular calcium on the cytoplasmic Ca2+ concentration (Ca2+i) were studied by dual-wavelength microfluorometry in individual human parathyroid cells obtained from adenomatous glands and normal-sized glands associated with adenomas in hypercalcemic hyperparathyroidism (HPT), as well as from enlarged glands of patients with uremia with HPT. In comparison with the normal parathyroid tissue, chief cells of the adenomatous and hyperplastic glands showed significantly lower Ca2+, and also right-shifted responses of Ca2+i to increases in the extracellular calcium concentration within the 0.5 to 3.0 mmol/L range. This pathophysiologic disturbance apparently was independent of the cell size. Oxyphil cells of nodules from the hyperplastic glands had lower Ca2+i and responded less to increments in extracellular Ca2+ than the chief cells from the surrounding parts of the same glands. Also the chief cells from the normal-sized glands associated with single adenomas exhibited a disturbance of the regulation of Ca2+i, which was less pronounced than that in the cells of the adenomas. These findings support the presence of relative calcium insensitivity of Ca2+i in chief and oxyphil parathyroid cells from adenomatous and hyperplastic glands. This derangement may also be found in all parathyroid glands of individuals with adenomatous HPT.


Gustatory sweating of the external auditory canal.
Gustatory sweating of the external auditory canal is extremely rare. A clinical case, that is only the second in the English literature, is presented. The potential pathogenesis and its treatment options are discussed.


Synchronous Pancreatic Serous Cystic Neoplasm and Duodenal Neuroendocrine Tumor: Case Report and Review of the Literature.
Simultaneous presence of pancreatic serous cystic neoplasms and neuroendocrine neoplasms is rare. We present a case with the incidental finding of a duodenal neuroendocrine tumor (NET) with 2 lymph node metastases in a Whipple resection specimen performed to remove a pancreatic cystic neoplasm that postoperatively turned out to represent a serous cystic neoplasm (SCN). The patient was a 75-year-old female. She presented with loss of appetite and weight. Preoperative contrast-enhanced computed tomography scan of the abdomen showed a multicystic lesion in the head of pancreas. On histologic examination of the resection specimen, a pancreatic SCN was found, and in addition in 2 peripancreatic lymph nodes, metastases from a NET. Further examination of the resected specimen revealed a duodenal NET. Review of the literature revealed only one prior study with 4 cases of pancreatic SCN associated with a duodenal NET. In 1 of the 4 cases, the patient had von Hippel-Lindau disease. Our report emphasizes the importance of careful examination of pancreatic resection specimens including the peripancreatic lymph nodes also when dealing with SCNs, as coexisting and more malignant tumors may otherwise be missed.


[Stomatological identification after air disasters].
The paper describes experiences of stomatological identification after two air disasters and shows potentialities and limits of an odontologic(al) comparison. The application of a 16-bit-computer is pointed out as the essential improvement of the work.


Adenovirus vector-mediated expression of TMEM166 inhibits human cancer cell growth by autophagy and apoptosis in vitro and in vivo.
TMEM166 is a novel programmed cell death-related molecule. In this report, we constructed a recombinant adenovirus 5-TMEM166 vector (Ad5-TMEM166) and evaluated its expression and anti-tumor activities in vitro and in vivo. Cell viability analysis revealed that the adenovirus-mediated increase of TMEM166 inhibited tumor cell growth in a dose- and time-dependent manner. This inhibitory effect was mediated by both autophagy (via inhibition of mTOR and activation of p70S6K) and apoptosis (via caspase-3 activation), both of which contributed to cell death and suppression of tumorigenicity. Our data indicated that Ad5-TMEM166 may be a novel gene therapy candidate for cancer.


Cell Type and Nuclear Size Dependence of the Nuclear Deformation of Cells on a Micropillar Array.
While various cellular responses to materials have been published, little concerns the deformation of cell nuclei. Herein we fabricated a polymeric micropillar array of appropriate dimensions to trigger the significant self-deformation of cell nuclei and examined six cell types, which could be classified into cancerous cells (Hela and HepG2) versus healthy cells (HCvEpC, MC3T3-E1, NIH3T3, and hMSC) or epithelial-like cells (Hela, HepG2, and HCvEpC) versus fibroblast-like cells (MC3T3-E1, NIH3T3, and hMSC). While all of the cell types exhibited severe nuclear deformation on the poly(lactide- co-glycolide) (PLGA) micropillar array, the difference between the epithelial-like and fibroblast-like cells was much more significant than that between the cancerous and healthy cells. We also examined the statistics of nuclear shape indexes of cells with an inevitable dispersity of nuclear sizes. It was found that larger nuclei favored more significant deformation on the micropillar array for each cell type. In the same region of nuclear size, the parts of the epithelial-like cells exhibited more significant nuclear deformation than those of the fibroblast-like cells. Hence, this article reports the nuclear size dependence of the self-deformation of cell nuclei on micropillar arrays for the first time and meanwhile strengthens the cell-type dependence.


Rapid-cycle testing cuts bed turnaround by 85%.
You can use rapid-cycle testing to try out new approaches to overcrowding much more frequently than with more traditional process improvement strategies. Improving bed turnaround notification can yield dramatic improvements. Telling staff they have to try a new process only for three days makes it easier to gain buy-in. Look for old policies that are no longer needed, yet continue to keep your staff bogged down.


Angiogenesis in peritoneal dialysis.
Long-term exposure to peritoneal dialysis fluid induces morphological alterations, including angiogenesis, leading to a loss of ultrafiltration (UF) capacity. We discuss the effect of different factors in peritoneal dialysis (PD) on angiogenesis. In addition, we describe the process of angiogenesis and the possible role of different cell types in the peritoneum upon PD contributing to new blood vessel formation. Furthermore, we review several interventions used in our rat PD exposure model to decrease angiogenesis in PD. Moreover, we show new data on the use of sunitinib to inhibit angiogenesis in this rat model. Although various interventions seem to be promising, well-randomised clinical trials showing absolute prevention of angiogenesis and UF failure are, yet, still missing. To make real progress in PD treatment, the aim should be to prevent angiogenesis as well as peritoneal fibrosis and PD-induced inflammation.


Quantitative aspects of analyzing small molecules - monitoring singly or doubly charged ions? A case study of ximelagatran.
Precision, reproducibility and lower limit of quantitation (LLOQ) are important characteristics of a quantitative method. We have investigated these properties for Ximelagatran (Xi), which has a high tendency to form doubly charged ions in electrospray ionization (ESI), by studying the percentage of doubly charged species formed when varying the formic acid (FA) concentration, analyte concentration, amount of organic modifier and flow rate. It was found that the percentage of [Xi + 2H](2+) can be controlled to be more than 90% or less than 10% by varying the amount of FA present, and that the change between these values is dramatic. Furthermore, the percentage of [Xi + 2H](2+) formed decreases with increased analyte concentration and increased flow rate. No apparent relationship with the amount of organic modifier was found. The results have the implication that, by carefully controlling the selected parameters, the LLOQ, precision and reproducibility can be improved. We have compared the fragmentation of the singly and doubly charged species and concluded that the [Xi + 2H](2+) ion is more inclined to undergo fragmentation than [Xi + H](+). As a consequence, unusual instrumental settings had to be used for the experiments. The fragmentation patterns are to a great extent similar, but the doubly charged species is more inclined to generate low-mass product ions.


The vasopressin-sensitive adenylate cyclase in collecting tubules and in thick ascending limb of Henle's loop of human and canine kidney.
The major tubular effects of [8-Arg]vasopressin (AVP) in regulation of renal water excretion are initiated by stimulation of adenylate cyclase (AdC) coupled with V2 receptors. We explored whether the AVP-sensitive AdC is present in both collecting tubules and the thick ascending limb of Henle's loop of human and canine kidney. In cortical collecting tubule (CCT) and medullary collecting tubules (MCT) of human kidney, AdC was markedly stimulated by AVP [maximum change from basal level (delta), +2700%] and the the nonhormonal stimulatory agent forskolin (delta, +2000%). In human CCT, the effects of both compounds were synergistic. In contrast, AVP had no effect on AdC in either the medullary (MAL) or cortical (CAL) segment of the thick ascending limb of Henle's loop of human kidney; AVP also did not stimulate AdC in CAL or MAL in the presence of forskolin. Similar to that in the human kidney, in the canine kidney, AdC in CCT and MCT was markedly stimulated by AVP and forskolin (delta, +1000%), but AVP had no effect on AdC in CAL and MAL of the canine kidney. In intact tubules dissected from dog kidney and incubated in vitro, AVP markedly increased cAMP accumulation in MCT. AVP also elicited a small but detectable increase in cAMP accumulation in MAL. From these observations, we conclude that AVP-sensitive AdC is well developed in collecting tubules, but that AVP-sensitive AdC is absent in MAL and CAL of human kidney. Likewise, in canine nephron, the AVP-sensitive AdC of MAL and CAL is rudimentary or very labile. These findings suggest that the unresponsiveness of the AdC-cAMP system to AVP in segments of the thick ascending limb of Henle's loop may be a factor that accounts for a relatively low maximum osmotic concentration of urine which can be achieved by human or canine kidneys.


Mapping evaporative water loss in desert passerines reveals an expanding threat of lethal dehydration.
Extreme high environmental temperatures produce a variety of consequences for wildlife, including mass die-offs. Heat waves are increasing in frequency, intensity, and extent, and are projected to increase further under climate change. However, the spatial and temporal dynamics of die-off risk are poorly understood. Here, we examine the effects of heat waves on evaporative water loss (EWL) and survival in five desert passerine birds across the southwestern United States using a combination of physiological data, mechanistically informed models, and hourly geospatial temperature data. We ask how rates of EWL vary with temperature across species; how frequently, over what areas, and how rapidly lethal dehydration occurs; how EWL and die-off risk vary with body mass; and how die-off risk is affected by climate warming. We find that smaller-bodied passerines are subject to higher rates of mass-specific EWL than larger-bodied counterparts and thus encounter potentially lethal conditions much more frequently, over shorter daily intervals, and over larger geographic areas. Warming by 4 °C greatly expands the extent, frequency, and intensity of dehydration risk, and introduces new threats for larger passerine birds, particularly those with limited geographic ranges. Our models reveal that increasing air temperatures and heat wave occurrence will potentially have important impacts on the water balance, daily activity, and geographic distribution of arid-zone birds. Impacts may be exacerbated by chronic effects and interactions with other environmental changes. This work underscores the importance of acute risks of high temperatures, particularly for small-bodied species, and suggests conservation of thermal refugia and water sources.


Processing of the 5'-UTR and existence of protein factors that regulate translation of tobacco chloroplast psbN mRNA.
The chloroplast psbB operon includes five genes encoding photosystem II and cytochrome b 6 /f complex components. The psbN gene is located on the opposite strand. PsbN is localized in the thylakoid and is present even in the dark, although its level increases upon illumination and then decreases. However, the translation mechanism of the psbN mRNA remains unclear. Using an in vitro translation system from tobacco chloroplasts and a green fluorescent protein as a reporter protein, we show that translation occurs from a tobacco primary psbN 5'-UTR of 47 nucleotides (nt). Unlike many other chloroplast 5'-UTRs, the psbN 5'-UTR has two processing sites, at -39 and -24 upstream from the initiation site. Processing at -39 enhanced the translation rate fivefold. In contrast, processing at -24 did not affect the translation rate. These observations suggest that the two distinct processing events regulate, at least in part, the level of PsbN during development. The psbN 5'-UTR has no Shine-Dalgarno (SD)-like sequence. In vitro translation assays with excess amounts of the psbN 5'-UTR or with deleted psbN 5'-UTR sequences demonstrated that protein factors are required for translation and that their binding site is an 18 nt sequence in the 5'-UTR. Mobility shift assays using 10 other chloroplast 5'-UTRs suggested that common or similar proteins are involved in translation of a set of mRNAs lacking SD-like sequences.


Quantitative structure-cytotoxicity relationship of newly synthesized tropolones determined by a semiempirical molecular-orbital method (PM5).
A semiempirical molecular-orbital method (CONFLEX/PM5) was applied to delineate the relationship between the cytotoxicity (evaluated by 50% cytotoxic concentration, CC(50)) of twenty-four tropolone-related compounds and their molecular weight or one of the following eleven chemical descriptors: the heat of formation (COSMO, non-COSMO; kcal/mole), stability of hydration (=COSMO-nonCOSMO (DeltaH); kcal/mole), dipole moment (D), hydrophobicity (log P), highest occupied molecular orbital energy (E(HOMO); eV), lowest unoccupied molecular orbital energy (E(LUMO); eV), absolute hardness [eta=(E(LUMO)-E(HOMO))/2; eV)], absolute electron negativity [chi=-(E(LUMO)+E(HOMO))/2; eV], reactivity index (omega=chi(2)/2eta; eV), surface area (A(2)) and volume (A(3)) of the molecule. No good correlation was found with the unseparated twenty-four compounds all together, but modest to high correlation was found after separation into three different groups of compounds, depending on the structural similarity. Particular descriptors could be used to evaluate the biological activity of newly synthesized tropolones.


[Australian method of first aid for European snake bites. Spreading its use in Italy].
Antiquated first-aid procedures, such as the incision and sucking of the wound and the application of a tourniquet, are still used in Italy consequent to the bite of vipers. The A. hope that such procedures will be finally abandoned, since they are ineffective, impractical, painful and can furthermore produce invalidating or disfiguring permanent outcomes. In this report they extensively describe Australian first-aid measures consequent to snakebites, constituted by a compressive bandage and by immobilization of the bitten limb by splinting. They hope therefore that the divulgation of this procedure in Italian will bring about, within a short period of time, its extensive use in Italy consequent to viper bites, as it is very effective, practical and easy to carry out.


Addressing the shortage. Strategies for building the nursing workforce.
For more than two decades, nursing educators and administrators have grappled with issues of declining student enrollments and the concurrent "defection" of nurses from the labor market. This article describes both short- and long-term strategies for addressing the nursing shortage and identifies new opportunities for collaboration in building the nursing workforce of the future.


Evaluation of Upper Limb Joint Contribution to Racket Head Speed in Elite Tennis Players Using IMU Sensors: Comparison between the Cross-Court and Inside-Out Attacking Forehand Drive.
This study aimed to quantify and compare the upper limb angular kinematics and its contributions to the racket head speed between the cross-court (CC) and inside-out (IO) attacking tennis forehand of elite tennis players in a competitive environment. A new approach was used to study the forehand drive with mini-inertial sensors of motion capture to record the kinematic data. Six strokes in each direction per participant (72 shots in total) were chosen for analysis. Upper limb kinematics were calculated in the Visual 3D platform (Visual 3D Professional V5.01.21, C-motion, Germantown, MD, USA). The method used to calculate the upper limb's contributions was performed with MATLAB software and used the segment's (upper arm, forearm and hand) angular velocities and their respective displacement vectors obtained through the inertial sensors. Upper limb kinematics demonstrated a higher shoulder rotation in the IO direction with significant differences at the end of the backswing, which could be a key factor in distinguishing the two directions of the shot. Results also demonstrated that the horizontal flexion of the upper arm (around the shoulder joint) was primarily responsible for the racket velocity in the anteroposterior direction (48.1% CC and 45.2% IO), followed by the extension of the forearm (around the elbow joint) (17.3% CC and 20.9% IO) and the internal rotation of the upper arm (around the shoulder joint) (15.6% CC and 14.2% IO). No significant differences were shown in the contributions of upper limbs to the racket head velocity between the two directions of the shot. Tennis coaches and players should develop a specific training programme to perform higher angular velocities in these specific joint rotations.


Sex-changing fish as a manipulable system for the study of the determination, differentiation, and stability of sex in vertebrates.
Two fundamental questions concerning vertebrate sexuality are what controls the stability of gender throughout the life of an individual and what are the genetic, immunological, and endocrine factors controlling sex determination and differentiation. Significant aspects of both issues cannot readily be examined experimentally in standard laboratory vertebrates partly because these species are gonochores and provide no opportunity to examine plasticity in sexual systems, and partly because few means have yet been found experimentally to manipulate immunogenetic factors, such as sex-specific DNA or cell-surface antigens, thought to be involved in the determination and differentiation of sex. Behaviorally induced adult sex change in hermaphroditic fishes constitutes a unique system of controlled sexual plasticity in which these issues can be addressed. Several species of tropical marine fishes satisfy the requirements for an appropriate laboratory experimental animal. Sex change in them is known to involve alterations in external coloration, behavior, gonadal structure, hormonal enzyme activity, levels of circulating steroid hormones, and concentrations of H-Y antigen. Genetic, immunological, endocrine, and behavioral factors interact with one another causally in ways permitting relatively simple manipulations simultaneously to analyze the network of sequential causes for sex change and to address the above-stated fundamental issues concerning sexuality generally in vertebrates. Since sex change can be started at will, these fish species become powerful model systems for the analysis of basic mechanisms of sex determination, differentiation, and long-term stability.


Positron emission tomography and magnetic resonance imaging in primary central nervous system lymphoma-a narrative review.
This review addresses the challenges of primary central nervous system (CNS) lymphoma diagnosis, assessment of treatment response, and detection of recurrence. Primary CNS lymphoma is a rare form of extra-nodal non-Hodgkin lymphoma that can involve brain, spinal cord, leptomeninges, and eyes. Primary CNS lymphoma lesions are most commonly confined to the white matter or deep cerebral structures such as basal ganglia and deep periventricular regions. Contrast-enhanced magnetic resonance imaging (MRI) is the standard diagnostic modality employed by neuro-oncologists. MRI often shows common morphological features such as a single or multiple uniformly well-enhancing lesions without necrosis but with moderate surrounding edema. Other brain tumors or inflammatory processes can show similar radiological patterns, making differential diagnosis difficult. [18F]-fluorodeoxyglucose (FDG) positron emission tomography (PET) has selected utility in cerebral lymphoma, especially in diagnosis. Primary CNS lymphoma can sometimes present with atypical findings on MRI and FDG PET, such as disseminated disease, non-enhancing or ring-like enhancing lesions. The complementary strengths of PET and MRI have led to the development of combined PET-MR systems, which in some cases may improve lesion characterization and detection. By highlighting active developments in this field, including advanced MRI sequences, novel radiotracers, and potential imaging biomarkers, we aim to spur interest in sophisticated imaging approaches.


Hydroxyapatite Biobased Materials for Treatment and Diagnosis of Cancer.
Great advances in cancer treatment have been undertaken in the last years as a consequence of the development of new antitumoral drugs able to target cancer cells with decreasing side effects and a better understanding of the behavior of neoplastic cells during invasion and metastasis. Specifically, drug delivery systems (DDS) based on the use of hydroxyapatite nanoparticles (HAp NPs) are gaining attention and merit a comprehensive review focused on their potential applications. These are derived from the intrinsic properties of HAp (e.g., biocompatibility and biodegradability), together with the easy functionalization and easy control of porosity, crystallinity and morphology of HAp NPs. The capacity to tailor the properties of DLS based on HAp NPs has well-recognized advantages for the control of both drug loading and release. Furthermore, the functionalization of NPs allows a targeted uptake in tumoral cells while their rapid elimination by the reticuloendothelial system (RES) can be avoided. Advances in HAp NPs involve not only their use as drug nanocarriers but also their employment as nanosystems for magnetic hyperthermia therapy, gene delivery systems, adjuvants for cancer immunotherapy and nanoparticles for cell imaging.


Identification of new cactus alkaloids in Backebergia militaris by tandem mass spectrometry.
Tandem mass spectrometry, applied to simple extracts of Backebergia militaris, indicated the presence of a number of new alkaloids, including fully aromatic oxygenated isoquinolines, their di- and tetra-hydro analogs, and beta-phenethylamines. These conclusions were supported by separation using radial tlc and comparison with authentic compounds. Traces of seven alkaloids new to this cactus species were identified; four (3,4,8,11) were known previously from other cacti. Three novel cactus alkaloids were identified and confirmed by synthesis as 7,8-dimethoxy-3,4-dihydroisoquinoline (12, dehydrolemaireocereine), 6,7-dimethoxyisoquinoline (13, backebergine), and 7,8-dimethoxyisoquinoline (14, isobackebergine). The last two compounds are the first simple, fully aromatic, isoquinoline alkaloids to be reported from the Cactaceae. The sensitivity of this approach to new alkaloid discovery is emphasized; the entire project consumed only 10 g of dried plant material.


Efficacy and safety of nebulized recombinant human DNase as rescue treatment for persistent atelectasis in newborns: case-series.
To evaluate the efficacy and safety of using recombinant human DNase (rhDNase) in diminishing persistent atelectasis unresponsive to conventional treatment and mucus plugging in newborns with insufficient ability to clear thick and purulent airway secretions. Twelve newborns (10 preterms), who did not respond to conventional methods, received rhDNase nebulized therapy at a dose of 1.25 mg over a 15-minute period, twice a day (2 hours between the doses) for up to 3 days. The application of the drug was continued for up to 3 days or until the improvement of atelectasis. After a-three-day therapy, if atelectasis did not improve, a single dose (1.25 mg) of the same drug in liquid form was administered endotracheally. Clinical (respiration rate, requirement for oxygen concentration) and radiological response (chest x-ray scoring), duration of the treatment, recurrence of atelectasis and requirement for additional therapy were evaluated. Ten out of 12 patients showed rapid clinical and radiological improvement after nebulized treatment. Two patients who did not respond to the three-day regimen received a single dose of the drug endotracheally and both recovered completely. Six patients did not require completion of three day regimen for radiological recovery. Chest x-ray scores and respiratory parameters showed significant improvement after the treatment. The respective median (range) values before and after treatment were 4 (1-5) and 0 (0-4) points for chest x-ray scores, 66 (60-78) and 49 (44-64) breaths/min for respiratory rates, and 45% (35-64) and 30% (21-40) for oxygen requirement. Comparison of pCO(2) before (median, 56 mm Hg; range, 46-64) and after treatment (median, 41 mm Hg; range 38-58) in 7 patients showed significant improvement. In a large series of newborns to receive rhDNase and we demonstrated the usefulness of rhDNase as a mucolytic agent in treating newborns with persistent atelectasis who do not respond to other treatments.


Cancer Mortality in Residents of the Cadmium-Polluted Jinzu River Basin in Toyama, Japan.
After 26 years, we followed up 7348 participants in a 1979-1984 health screening survey in the Jinzu River basin, the heaviest cadmium-polluted area in Japan. We assessed the associations of cadmium exposure levels and mortality from cancer and renal damage, indicated by records of proteinuria and glucosuria in the original survey. Mortality risks (hazard ratios) were analyzed using the Cox proportional hazards model, stratified by sex, after adjusting for age, smoking status, and hypertension, as indicated in the original survey records. In men, the adjusted hazard ratio for mortality from lung cancer was significantly lower in individuals residing in an area of historically high cadmium exposure and in subjects with a historical record of proteinuria, glucosuria, and glucoproteinuria. The risk of mortality from prostate cancer was borderline higher in cadmium-exposed men. In women, historical cadmium exposure was not associated with an increased risk of mortality from malignant neoplasms, but the adjusted hazard ratios for death from total malignant neoplasms or from renal and uterine cancers were significantly higher in exposed subjects with a historical record of proteinuria, glucosuria, and glucoproteinuria. These findings suggest that women residing in cadmium-polluted areas who exhibit markers of renal damage may be at risk of dying of cancer.


Nursing team knowledge on care for patients with fungating wounds.
to evaluate the nursing team knowledge of a cancer hospital on care for patients with Malignant Fungating Wounds (MFW) and to analyze associated sociodemographic and educational factors. an observational and cross-sectional study, conducted between September and October 2015, after approval by the Research Ethics Committee. A questionnaire was applied containing sociodemographic, educational and related components to the accomplishment of dressings, dressings choice and orientation. Data were analyzed by using Chi-square, Fisher's exact test, Student's t-Test and Pearson's correlation. 37 professionals participated in the study, most of whom were technicians (56.8%), women (91.9%) and with a mean age of 32 years. The professionals presented 56.5% of correct answers. There were no statistically significant associations between sociodemographic/educational variables and number of correct answers. there was a lack of important knowledge about care for patients with MFW, which should guide strategies for the oncology staff training.


A constitutive, transient receptor potential-like Ca2+ influx pathway in presynaptic nerve endings independent of voltage-gated Ca2+ channels and Na+/Ca2+ exchange.
Calcium levels in the presynaptic nerve terminal are altered by several pathways, including voltage-gated Ca(2+) channels, the Na(+)/Ca(2+) exchanger, Ca(2+)-ATPase, and the mitochondria. The influx pathway for homeostatic control of [Ca(2+)](i) in the nerve terminal has been unclear. One approach to detecting the pathway that maintains internal Ca(2+) is to test for activation of Ca(2+) influx following Ca(2+) depletion. Here, we demonstrate that a constitutive influx pathway for Ca(2+) exists in presynaptic terminals to maintain internal Ca(2+) independent of voltage-gated Ca(2+) channels and Na(+)/Ca(2+) exchange, as measured in intact isolated nerve endings from mouse cortex and in intact varicosities in a neuronal cell line using fluorescence spectroscopy and confocal imaging. The Mg(2+) and lanthanide sensitivity of the influx pathway, in addition to its pharmacological and short hairpin RNA sensitivity, and the results of immunostaining for transient receptor potential (TRP) channels indicate the involvement of TRPC channels, possibly TRPC5 and TRPC1. This constitutive Ca(2+) influx pathway likely serves to maintain synaptic function under widely varying levels of synaptic activity.


The prevalence of bronchiectasis in patients with Marfan syndrome.
An association has been suggested between Marfan syndrome (MFS) and the nodular bronchiectatic form of lung disease caused by non-tuberculous mycobacteria (NTM). We evaluated the prevalence of bronchiectasis in 79 adult patients with MFS using computed tomography (CT) imaging. Airway dilation indicative of bronchiectasis (22/79, 28%) and bronchioloectasis (10/79, 13%) were relatively common, although the extent of dilation was not severe and was frequently confined to a single lobe. However, bronchiolitis was evident in only three patients (4%), and no patient was diagnosed with NTM lung disease.


[A clinical application of the T-Scan II system--usefulness for evaluating occlusal contacts of complete denture wearers].
The purpose of this study was to assess the reproducibility of the T-Scan II system and its clinical usefulness for evaluating occlusal contacts of complete denture wearers. The occlusal contacts of 13 dentate subjects, and 14 complete denture wearers, were recorded using a T-Scan II system during maximum voluntary clenching. The recordings for complete denture wearers were taken after each treatment for the new denture, and continued until the completion of all corrections. The recordings for complete denture wearers were analyzed using repeated measured ANOVA. In addition, the variables obtained with the system, the delta of the occlusal area and load, and the maximum-load time (MLT), which represented the time length taken to reach the occlusal load at the maximum level, were then compared between dentate subjects and complete denture wearers, using a t-test. The conclusions were as follows: 1. The standard errors values for both occlusal area and the load recordings for the dentate subjects were limited to within 10% of the means. 2. The values of the occlusal area and load significantly increased, and delta of the occlusal area and load significantly decreased, as the denture corrections were repeated. 3. The means of the MLT were about 0.3 seconds for dentate subjects, and 0.8 seconds for complete denture subjects. It was revealed that the T-Scan II showed acceptable reproducibility, and it was useful to evaluate occlusal contacts of complete denture wearers.


Cytotoxicity and genotoxicity of sodium percarbonate: a comparison with bleaching agents commonly used in discoloured pulpless teeth.
To evaluate the cytotoxicity and genotoxicity of sodium percarbonate (SPC) in comparison with bleaching agents used on discoloured pulpless teeth. The cytotoxicity and genotoxicity of bleaching agents were evaluated both in their pure form as well as at concentrations commonly used in clinical practice. Hydrogen peroxide (HP), carbamide peroxide (CP), sodium perborate (SP) and SPC were diluted in Dulbecco's modified Eagle's medium (DMEM) in series. To evaluate the cytotoxicity, the survival of 3T3/NIH mouse fibroblasts was measured photometrically using an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay after a 24 h-exposure period. Genotoxicity was indicated by micronuclei (MN) formation, and modification of the normal cell was analysed by light microscopy (400x). Statistical analysis was performed by one-way anova, followed by a multiple-comparison Tukey post hoc test (P < 0.05). All groups exhibited a dose-dependent cytotoxicity. However, CP showed a similar cytotoxic effect when compared with DMEM-untreated control (UC) group. HP and SPC were significantly more cytotoxic than SP. The genotoxicity test showed that SPC and SP had an intermediate rate of MN frequency when compared with the UC group. The mean rate of MN frequency for HP was higher and statistically more significant than for the other groups tested. No difference was observed when CP and UC groups were compared. Sodium percarbonate showed cytotoxicity and genotoxicity similar to those of the other products tested. However, before SPC is used clinically, studies should be conducted to confirm its safety in vivo.


Study of 500 patients with limb joint osteoarthritis. I. Analysis by age, sex, and distribution of symptomatic joint sites.
Five hundred subjects with symptomatic limb joint osteoarthritis, who had been referred to a rheumatologist, were enrolled into a continuing study. They comprised 342 women (mean age 65.3) and 158 men (mean age 59.7), with a mean symptom duration of 15.4 years at entry. Only 31 patients (6%) had symptomatic osteoarthritis of one joint alone; however, in a further 205 (41%) the disease was limited to one site. One hundred and eighty two (36.4%) had two sites affected and 82 (16.4%) three or more sites of symptomatic osteoarthritis. Of 847 affected joints the most commonly involved were 349 (41.2%) knees, 254 (30%) hands, and 161 (19%) hips. Hip disease stood out as a separate entity, often occurring alone, and having a stronger male preponderance and different associations than osteoarthritis at other joint sites. Knee and hand disease were significantly associated in women. Obesity, hypertension, and Heberden's nodes were common. The number of sites affected, as well as the distribution, was strongly related to age as well as sex, suggesting that polyarticular osteoarthritis arises from slow acquisition of new joint sites in a non-random distribution. 'Generalised' osteoarthritis did not emerge as a distinct entity.


Prostate-specific membrane antigen for prostate cancer theranostics: from imaging to targeted therapy.
In recent years, major advances in molecular imaging of prostate cancers (PCa) were made with the development and clinical validation of highly accurate PET tracers to stage and restage the disease. Prostate-specific membrane antigen (PSMA) is a transmembrane protein highly expressed in PCa, and its expression has led to the development of PSMA-binding radiopharmaceuticals for molecular imaging or radioligand therapy (RLT). We herein review the recent literature published on diagnostic and therapeutic (i.e. theranostic) PSMA tracers. Development in small PSMA-targeted molecules labeled with gallium-68 and fluorine-18 show promising results for primary staging and detection of disease at biochemical recurrence using PET/computed tomography (PET/CT). Studies show a higher sensitivity and specificity, along with an improved detection rate over conventional imaging (CT scan and bone scan) or choline PET tracers, especially for restaging after prostate-specific antigen failure following loco-regional therapy. In addition, some PSMA tracers can be labeled with beta-minus and alpha particle emitters, yielding encouraging response rates and low toxicity, and potentially offering a new line of targeted therapy for metastatic castration-resistant PCa. PSMA-targeted tracers have shown unprecedented accuracy to stage and restage PCa using PET/CT. Given their specific biodistribution toward PCa tissue, PSMA RLT now offers new therapeutic possibilities to target metastatic PCa. Prospective multicenter randomized studies investigating the clinical impact of PSMA-targeted molecules are urgently needed.


Strange attractors and their periodic repetition.
In this paper, we present some important findings regarding a comprehensive characterization of dynamical behavior in the vicinity of two periodically perturbed homoclinic solutions. Using the Duffing system, we illustrate that the overall dynamical behavior of the system, including strange attractors, is organized in the form of an asymptotic invariant pattern as the magnitude of the applied periodic forcing approaches zero. Moreover, this invariant pattern repeats itself with a multiplicative period with respect to the magnitude of the forcing. This multiplicative period is an explicitly known function of the system parameters. The findings from the numerical experiments are shown to be in great agreement with the theoretical expectations.


An Eighteen-Month Helicobacter Infection Does Not Induce Amyloid Plaques or Neuroinflammation in Brains of Wild Type C57BL/6J Mice.
There is increasing evidence to support the role of infectious agents in the progression of Alzheimer's disease (AD), especially Helicobacter pylori (H. pylori). The impact of Helicobacter infection on the brain of non-AD predisposed mice was studied. For that, C57BL/6J mice were infected by oral gavage with H. pylori SS1 (n = 6) and Helicobacter felis (H. felis) (n=6) or not infected (n = 6) for evaluation of neuroinflammation (anti-GFAP and anti-iba1 immunohistochemistry) and amyloid-β deposition (thioflavin-S stain and anti-Aβ immunohistochemistry). After 18-month of infection, H. pylori SS1 and H. felis infection induced a strong gastric inflammation compared to non-infected mice, but did not induce brain neuroinflammation or amyloid-β deposition.


MAPkinases regulate secondary metabolism, sexual development and light dependent cellulase regulation in Trichoderma reesei.
The filamentous fungus Trichoderma reesei is a prolific producer of plant cell wall degrading enzymes, which are regulated in response to diverse environmental signals for optimal adaptation, but also produces a wide array of secondary metabolites. Available carbon source and light are the strongest cues currently known to impact secreted enzyme levels and an interplay with regulation of secondary metabolism became increasingly obvious in recent years. While cellulase regulation is already known to be modulated by different mitogen activated protein kinase (MAPK) pathways, the relevance of the light signal, which is transmitted by this pathway in other fungi as well, is still unknown in T. reesei as are interconnections to secondary metabolism and chemical communication under mating conditions. Here we show that MAPkinases differentially influence cellulase regulation in light and darkness and that the Hog1 homologue TMK3, but not TMK1 or TMK2 are required for the chemotropic response to glucose in T. reesei. Additionally, MAPkinases regulate production of specific secondary metabolites including trichodimerol and bisorbibutenolid, a bioactive compound with cytostatic effect on cancer cells and deterrent effect on larvae, under conditions facilitating mating, which reflects a defect in chemical communication. Strains lacking either of the MAPkinases become female sterile, indicating the conservation of the role of MAPkinases in sexual fertility also in T. reesei. In summary, our findings substantiate the previously detected interconnection of cellulase regulation with regulation of secondary metabolism as well as the involvement of MAPkinases in light dependent gene regulation of cellulase and secondary metabolite genes in fungi.


Extraction efficiency of shosaikoto (xiaochaihu tang) and investigation of the major constituents in the residual crude drugs.
Shosaikoto (Xiaochaihu Tang) is one of the frequently used traditional herbal medicines (Kampo medicines, Japan). To realize the effective use of precious crude drugs, we investigated the major constituents in the residual crude drugs after decoction and aimed for extraction efficiency of shosaikoto with regard to the extract, tannin content, and major constituents. We found that the residual crude drugs had large amounts of compounds, especially saikosaponin b(2), which had a 78.3% yield compared to that in the first decoction. The extraction efficiency increased when decoction time and volume of water increased. Both increases had an additive effect on the yield of the extract and saikosaponin b(2) in particular. We also found that the size of crude drug pieces that are available in Japanese markets is suitable for decoction because of quick permeation of water. From our study, the second decoction may be a valuable contribution to medical treatment and of effective use of crude drugs. Moreover, time and volume of water should be increased when patients have trouble in preparing a decoction. Our study revealing the factors that influence the extraction efficiency of shosaikoto will be the basis for empirical evidence about decocting Kampo medicine.


Local linear model and restoration method of underwater images.
When light transports in water, it will be scattered and absorbed by the water body and water particles, resulting in blurred images and color distortion. In order to improve the quality of underwater imaging, the local linear model and restoration method of underwater images are proposed in this paper. Based on the distance-invariant feature in the local region, the local linear model is established, and the slope and intercept of the model represent the transmission rate and the backscattered light of the local region of the image, respectively. Utilizing this model, the problem of underwater image restoration has been transformed into the problem of solving the slope and intercept of linear equations. To solve the linear imaging model, the concept of local special-value is defined in this paper, and several fitting points can be obtained through the special-value. Then the linear model is solved by the fitting method, and the restoration of underwater images is completed. The restoration results of different underwater scene images verify that the linear model has a good effect in improving the image clarity and removing the color distortion.


Chloroplast Protein 12 Expression Alters Growth and Chilling Tolerance in Tropical Forage Stylosanthes guianensis (Aublet) Sw.
Stylosanthes guianensis (Aublet) Sw. is a tropical forage legume with soil acidity tolerance and excellent adaptation to infertile soils, but sensitive to chilling. To understand the molecular responses of S. guianensis to chilling, differentially expressed genes between a chilling tolerant mutant 7-1 and the wild type were identified using suppression subtractive hybridization, and eight of them were confirmed and the regulation pattern were analyzed using quantitative reverse transcription PCR (qRT-PCR). Chloroplast protein 12 (CP12) functions to regulate the Calvin cycle by forming a ternary complex with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and phosphoribulokinase (PRK). SgCP12 transcript was induced by chilling in both plants, and higher levels were observed in 7-1 than in the wild type, implying a potential role of SgCP12 in chilling tolerance. To confirm this, transgenic S. guianensis plants over-expressing or down-regulating SgCP12 were generated, respectively. Higher Fv/Fm and survival rate and lower ion leakage were observed in transgenic plants overexpressing SgCP12 as compared with the wild type after chilling treatment, while lower Fv/Fm and survival rate and higher ion leakage were found in SgCP12 antisense plants. SgCP12 overexpression plants showed promoted growth with increased plant height and fresh weight, while the antisense plants exhibited reduced growth with decreased plant height and fresh weight as compared with the wild type. The results indicated that regulation of SgCP12 expression alters plant growth and chilling tolerance in S. guianensis. In addition, higher levels of net photosynthetic rate (Pn), GAPDH and PRK activities were observed in SgCP12 overexpression transgenic plants, while lower levels in antisense plants than in the wild type under both control and chilling conditions, indicating that altered activities of GAPDH and PRK were associated with the changed Pn in transgenic S. guianensis. Our results suggest that SgCP12 regulates GAPDH and PRK activities, Pn, and chilling tolerance in S. guianensis.


Anaerobic mineralization of quaternary carbon atoms: isolation of denitrifying bacteria on pivalic acid (2,2-dimethylpropionic acid).
The degradability of pivalic acid was established by the isolation of several facultative denitrifying strains belonging to Zoogloea resiniphila, to Thauera and Herbaspirillum, and to Comamonadaceae, related to [Aquaspirillum] and Acidovorax, and of a nitrate-reducing bacterium affiliated with Moraxella osloensis. Pivalic acid was completely mineralized to carbon dioxide. The catabolic pathways may involve an oxidation to dimethylmalonate or a carbon skeleton rearrangement, a putative 2,2-dimethylpropionyl coenzyme A mutase.


The respective contribution of cognitive control and working memory to semantic and subjective organization in aging.
Organizing information is beneficial to episodic memory performance. Among several possible organizational strategies, two consist of organizing the information in semantic clusters (semantic organization) or self-organizing the information based on new associations that do not exist in semantic memory (subjective organization). Here, we investigated in a single study how these two organizational behaviors were underlined by different controlled processes and whether these relations were subjected to age-related differences. We tested 123 younger adults (n = 63) and older adults (n = 60) on two episodic memory tasks, one where the words were organizable and another where the words were not organizable, allowing for semantic and subjective organization, respectively. Additionally, participants were tested on three cognitive control tasks (Wisconsin Card Sorting Test, Stroop Test and Trail Making Test) and three working memory tasks (Backward Digit Span, Alpha Span and N-back test). Results revealed well-established age-related differences in terms of recall performance and organizational strategy implementation. More importantly, we found evidence that the different cognitive tests statistically yielded two different latent factors, a cognitive control factor and a working memory factor. Based on this dissociation, we found that only cognitive control contributed to semantic organization in all age groups whereas only working memory contributed to subjective organization, also in all age groups. These results shed new lights on our understanding of how controlled processes differently contribute to organizational behaviors in episodic memory. (PsycInfo Database Record (c) 2023 APA, all rights reserved).


Autologous fat injection to treat leakage around tracheoesophageal puncture.
To evaluate prospectively autologous fat injection as a treatment for leakage around tracheoesophageal puncture. For ten patients who exhibited leakage of saliva or liquid around their tracheoesophageal puncture, autologous fat injection was carried out into the tracheoesophageal wall around the maintained prosthesis. Short-term success was evaluated at one month, and long-term follow-up was continued as long as the patients remained alive. The effectiveness of the procedure was evaluated based on the presence or absence of leakage around the puncture as the patient swallowed methylene blue liquid. Patients were adjudged completely improved when no leakage of blue liquid was observed, partially improved when a slight stasis was evident, or unimproved when a marked leakage of blue liquid was noted. Short-term success was achieved for 6 patients, 2 patients partially improved, and the procedure failed for 2 others. Long-term success (from 10 to 65 months) was achieved in all 4 of the completely improved patients who were free of disease, including 1 patient who required an additional injection. Removal of the puncture was later carried out in the 2 others for cervical node metastases or tracheal tumor. Definitive or transitional puncture closure or removal of the prosthesis was required in 3 partially or unimproved patients, whereas 1 partially improved patient died of lung metastasis 3 months later. Fat injection around a voice prosthesis is a procedure that may be conducted as an alternative to other conservative techniques to decrease the size of the puncture. Close endoscopic observation is required however to eliminate secondary tumor that might expand the size of the puncture.


Molecular cloning of the common acute lymphoblastic leukemia antigen (CALLA) identifies a type II integral membrane protein.
Common acute lymphoblastic leukemia antigen (CALLA) is a 100-kDa cell-surface glycoprotein expressed on most acute lymphoblastic leukemias and certain other immature lymphoid malignancies and on normal lymphoid progenitors. The latter are either uncommitted to B- or T-cell lineage or committed to only the earliest stages of B- or T-lymphocyte maturation. To elucidate to homogeneity, obtained the NH2-terminal sequence from both the intact protein and derived tryptic and V8 protease peptides and isolated CALLA cDNAs from a Nalm-6 cell line lambda gt10 library using redundant oligonucleotide probes. The CALLA cDNA sequence predicts a 750-amino acid integral membrane protein with a single 24-amino acid hydrophobic segment that could function as both a transmembrane region and a signal peptide. The COOH-terminal 700 amino acids, including six potential N-linked glycosylation sites compose the extracellular protein segment, whereas the 25 NH2-terminal amino acids remaining after cleavage of the initiation methionine form the cytoplasmic tail. CALLA+ cells contain CALLA transcripts of 2.7 to 5.7 kilobases with the major 5.7- and 3.7-kilobase mRNAs being preferentially expressed in specific cell types.