ESR study of development of RFM/Un murine myeloid leukaemia.
The blood, spleen and liver of RFM/Un mice were examined by means of electron spin resonance (ESR) throughout the course of myeloid leukaemia induced by i.v. injection of leukaemic spleen cells. Marked changes in the concentration of iron transferrin and caeruloplasmin were observed in the blood 1 day after injection. As the disease progressed, changes occurred in the concentrations of the ascorbyl radical and of paramagnetic metal complexes in both spleen and liver. These changes are compared with those induced in RF/J mice injected with normal and leukaemic spleen cells from RFM/Un mice.


Facilitators and Barriers of Human Milk Donation.
The purpose of this study is to examine perceptions and experiences of women who donate human milk and highlight various aspects of the breast milk donation process. A cross-sectional descriptive study. An online survey was conducted with a convenience sample of women who donated milk at several milk banks in the United States. A questionnaire of 36 closed and open-ended items were developed and validated by the research team. Descriptive statistics and content analysis were used. Semantic content analysis involved three procedures: coding, categorizing text units, and refining the identified themes. A total of 236 women who donated breast milk completed the questionnaire. Mean age of participants was 32.7±4.27 and 89.40% were non-Hispanic White women with a bachelor's degree (32.20%) or graduate degree (54.70%). Most participants were women who actively donated breast milk, ranging from one to four times. Two themes, facilitators and barriers of milk donation, were identified. Facilitators to milk donation included attitudes toward milk donation, commitment for donating, motivation in donating, and support. Barriers included personal factors, environment, milk donor process, and psychosocial factors. Nurses, health care providers, and lactation professionals should educate women about milk donation resources and opportunities. Strategies to increase awareness about milk donation among underrepresented groups such as women of color are highly recommended. Future research is needed to further explore specific factors that increase milk donation awareness and minimize barriers to potential donors.


State-dependent TMS reveals the differential contribution of ATL and IPS to the representation of abstract concepts related to social and quantity knowledge.
The neural representation of abstract concepts is a current matter of debate. While social concepts have been associated to superior anterior temporal lobe (sATL), the neural correlates of quantity-related concepts have seldom been investigated. The right intraparietal sulcus (IPS) is known to be involved in the processing of numerical information and of quantifiers. In the present study, we investigated the causal role of right sATL and right IPS in representing social and quantity-related concepts, using a state-dependent transcranial magnetic stimulation (TMS)-priming paradigm. We modulated the neural activation by priming to a category label (either SOCIAL or QUANTITY), before applying TMS and presenting the target (i.e., an exemplar of one of the two abstract classes). When the target word belonged to the quantity category, TMS applied over the IPS (but not sATL) abolished the expected priming effect by speeding up reaction times (RTs) in incongruent trials. For the social category, both IPS and sATL stimulation abolished the priming effect, by reducing RTs in incongruent trials. Our results suggest a specialization of distinct brain areas in processing different classes of abstract concepts. The right IPS contains neuronal representations tuned to quantity-related concepts, in line with its well-known role in numerical and magnitude representation. Social concepts, in contrast, are represented in both the right sATL and the right IPS, known to be involved in social cognition and person-related knowledge.


Inhibition of human chondrosarcoma cell growth via apoptosis by peroxisome proliferator-activated receptor-gamma.
A rare immunohistochemical study using 28 surgical sections of human chondrosarcoma revealed that 67.9% of tumour cells had weak (10-40%) or strong (>40%) positive immunoreaction for peroxisome proliferator-activated receptor-gamma. The expression of peroxisome proliferator-activated receptor-gamma mRNA and protein in human chondrosarcoma cell line OUMS-27 was also determined by reverse transcription-polymerase chain reaction and immunocytochemistry, respectively. Furthermore, the effects of peroxisome proliferator-activated receptor-gamma ligands on cell proliferation and survival were investigated in OUMS-27 cells. Pioglitazone, a selective ligand for peroxisome proliferator-activated receptor-gamma, and 15-deoxy-Delta(12,14)-prostaglandin J(2) (15d-PGJ(2)), a putative endogenous ligand for peroxisome proliferator-activated receptor-gamma, inhibited the proliferation of OUMS-27 cells in a dose-dependent manner. The mechanism of cytotoxic effects of 15d-PGJ(2) was via apoptosis as shown by DNA fragmentation using TUNEL stain and DNA ladder formation, and by ultrastructural analysis using transmission electron microscopy. Flow-cytometric analysis using annexin-V-fluorescein and propidium iodide detected the early change of apoptosis, as well as necrosis of OUMS-27 cells at 4 h after co-incubation with 15d-PGJ(2). These results suggest that peroxisome proliferator-activated receptor-gamma may play a significant role in the pathogenesis of chondrosarcoma, and peroxisome proliferator-activated receptor-gamma ligands, especially 15d-PGJ(2), may be of therapeutic value in the treatment of human chondrosarcoma.


The Lathyrus sativus neurotoxin: resistance of the squirrel monkey to prolonged oral high doses.
Prolonged oral administration of the Lathyrus sativus neurotoxin, L-3-oxalylamino-2-aminopropionic acid (OAP), to three young male squirrel monkeys at dose rates of 0.6 to 6.0 mg/g body weight/day produced no neurologic signs and no adverse effects other than depressed activity and occasional foaming at the mouth. When the dose was increased to 8.0 to 8.5 mg/kg body weight/day in two animals, seizures and death occurred after 3 and 5 days. The signs were typical of acute OAP intoxication observed previously to occur in the squirrel monkey within an hour after a single ip dose (2 mg/g body weight) of OAP. Total cumulative dosage of OAP and duration of the experiments were as follows: Experiment 1, 9.5 g, 38 days; Experiment 2, 157 g, 178 days; Experiment 3, 64 g, 33 days. Histologic examination revealed no abnormalities in brain, spinal cord, or other organs. These experiments suggest that the adult squirrel monkey is highly resistant to chronic oral OAP intoxication. Under our experimental conditions, this species did not provide a satisfactory animal model for human neurolathyrism, a disease postulated to result from chronic OAP intoxication.


Progress in metamodulation and receptor-receptor interaction: From physiology to pathology and therapy.
The organization and the role of receptor-receptor interaction (RRI) and metamodulation in physiological conditions have been extensively analyzed and discussed. In this Special Issue of Neuropharmacology, we review recent advances in the understanding of the RRI and the mechanisms underlying its adaptation that could be relevant to the etiopathogenesis of central neuropsychiatric disorders, as well as to the development of new therapeutic approaches to control the activity and to restore the physiological functions, posing the basis for new targeted pharmacological interventions.


ST2 blockade reduces sST2-producing T cells while maintaining protective mST2-expressing T cells during graft-versus-host disease.
Graft-versus-host disease (GVHD) remains a devastating complication after allogeneic hematopoietic cell transplantation (HCT). We previously identified high plasma soluble suppression of tumorigenicity 2 (sST2) as a biomarker of the development of GVHD and death. sST2 sequesters interleukin-33 (IL-33), limiting its availability to T cells expressing membrane-bound ST2 (mST2) [T helper 2 (TH2) cells and ST2(+)FoxP3(+) regulatory T cells]. We report that blockade of sST2 in the peritransplant period with a neutralizing monoclonal antibody (anti-ST2 mAb) reduced GVHD severity and mortality. We identified intestinal stromal cells and T cells as major sources of sST2 during GVHD. ST2 blockade decreased systemic interferon-γ, IL-17, and IL-23 but increased IL-10 and IL-33 plasma levels. ST2 blockade also reduced sST2 production by IL-17-producing T cells while maintaining protective mST2-expressing T cells, increasing the frequency of intestinal myeloid-derived suppressor cells, and decreasing the frequency of intestinal CD103 dendritic cells. Finally, ST2 blockade preserved graft-versus-leukemia activity in a model of green fluorescent protein (GFP)-positive MLL-AF9 acute myeloid leukemia. Our findings suggest that ST2 is a therapeutic target for severe GVHD and that the ST2/IL-33 pathway could be investigated in other T cell-mediated immune disorders with loss of tolerance.


Application of a novel second-order differential equation grey model to forecast NOx emissions in China.
Nitrogen oxide (NOx) contains two harmful air pollutants: nitric oxide (NO) and nitrogen dioxide (NO2). The reasonable prediction of China's NOx emissions is of positive significance for the government to formulate environmental protection policies. To this end, a new grey prediction model with second-order differential equation is proposed in this paper, which has more reasonable model structure and better modeling performance than the traditional grey model. Secondly, according to the data characteristics of NOx emissions of China in recent years, a smoothing algorithm and weakening buffer operator are employed to process the original data to solve the rationality of the prediction results of the new model. Thirdly, the model for predicting China's NOx emissions has been constructed by the new proposed model. The results show that the mean comprehensive error of the new model is only 0.0692%, and its performance is much better than that of several other mainstream grey prediction models. Finally, the new model is applied to China's carbon dioxide prediction in the next 5 years, and the rationality of the prediction results is analyzed. Based on the prediction results, relevant countermeasures and suggestions are put forward.


Radioimmunoassay of methotrexate: use of 75Se-labelled methotrexate.
75Se-labelled methotrexate has been used as the radiolabel in a radioimmunoassay for methotrexate and its use compared with that of 3H-methotrexate. The gamma-labelled drug has proved to be extremely reliable and easier to use than 3H-methotrexate. Results obtained with it compare well with those obtained with tritiated drug. There are practical advantages in using a gamma emitter, and it is suggested that the modified radioimmunoassay is invaluable for routine monitoring of methotrexate in patients undergoing chemotherapy with the drug.


Vitellogenesis in Drosophila: sequestration of a yolk polypeptide/invertase fusion protein into developing oocytes.
The mechanism of yolk deposition into developing oocytes of Drosophila was investigated by following the fate of a reporter protein fused to a vitellogenin, or yolk polypeptide (YP). Embryos were transformed with a hybrid gene consisting of the promotor and amino terminal 430 codons of the Yp2 gene fused to the cytoplasmic form of the invertase gene from the yeast Saccharomyces cerevisiae. RNA hybridization experiments with established lines of transformed flies showed that the hybrid gene was expressed in female fat bodies and ovaries but not in any male cells. Immunoblotting and endoglycosidase digestion showed that the hybrid protein was secreted from fat body cells via the secretory pathway, transported in hemolymph, and sequestered into developing oocytes. Transfusion experiments with hemolymph and pure invertase showed that sequestration of invertase depended on its attachment to YP. Immunocytochemistry demonstrated that the hybrid protein became localized in yolk granules as oocytes developed. Females homozygous for the fusion gene are generally sterile; their eggs containing the hybrid protein often collapse and their embryos fail to develop, suggesting that the structure of the yolk polypeptides is important for embryonic development. These experiments show that YP2 carries structural information sufficient to direct a reporter protein from fat body cells, through the hemolymph, and into the yolk granules of developing oocytes. This work provides a means of identifying the features of yolk polypeptides that are responsible for their deposition into yolk during oogenesis.


Frequency and fidelity of translesion synthesis of site-specific N-2-acetylaminofluorene adducts during DNA replication in a human cell extract.
We have previously analyzed the effects of site-specific N-2-acetylaminofluorene (AAF) adducts on the efficiency and frameshift fidelity of SV40-based DNA replication in a human cell extract (Thomas, D. C., Veaute, X., Kunkel, T. A., and Fuchs, R. P. P. (1994) Proc. Natl. Acad. Sci. U.S.A. 91, 7752-7756). Here we use two sets of substrates to examine the probability of replication termination and error-free and error-prone bypass of AAF adducts. The substrates contained site-specific adducts at one of three guanines in a NarI sequence (5'-GGCGCC-3') placed within the lacZ alpha reporter gene and located on the template for either leading or lagging strand replication. The presence of the adduct at any position strongly reduces the efficiency of a single round of replication in a HeLa cell extract. Product analysis reveals preferential replication of the undamaged strand and termination of replication of the damaged strand occurring one nucleotide before incorporation opposite either a leading or lagging strand adduct. Products resistant to restriction endonuclease cleavage at the adducted site were generated in amounts consistent with 16-48% lesion bypass during replication. Most of this bypass was error-free. However, two-nucleotide deletion errors were detected in the replication products of DNA containing an AAF adduct in either the leading or lagging strand, but only when present at the third guanine position. Collectively, the data suggest that the replication apparatus in a HeLa cell extract generates a template-primer slippage error at an AAF adduct once for every 30-100 bypass events.


Destroyed by Slavery? Slavery and African American Family Formation Following Emancipation.
This study introduces a new sample that links people and families across 1860, 1880, and 1900 census data to explore the intergenerational impact of slavery on black families in the United States. Slaveholding-the number of slaves owned by a single farmer or planter-is used as a proxy for experiences during slavery. Slave family structures varied systematically with slaveholding sizes. Enslaved children on smaller holdings were more likely to be members of single-parent or divided families. On larger holdings, however, children tended to reside in nuclear families. In 1880, a child whose mother had been on a farm with five slaves was 49 % more likely to live in a single-parent household than a child whose mother had been on a farm with 15 slaves. By 1900, slaveholding no longer had an impact. However, children whose parents lived in single-parent households were themselves more likely to live in single-parent households and to have been born outside marriage.


Cleaners among wrasses: Phylogenetics and evolutionary patterns of cleaning behavior within Labridae.
Cleaner fishes remove and consume ectoparasites and are often categorized by whether they perform this behavior: (1) predominately as juveniles, (2) facultatively throughout ontogeny, or (3) obligately. Through a literature search, we confirmed that with at least 58 species exhibiting cleaning behavior, the Labridae (wrasses, parrotfishes, and allies) contain the highest diversity of cleaner fishes. In fact, there are 3-4 times as many cleaners within labrids as there are in any other marine group. The distribution and underlying causes of this exceptional diversity have not been determined. Here, we assess the topological and temporal patterns of labrid cleaner evolution. We used maximum likelihood and Bayesian approaches to infer the phylogenetic relationships and divergence times between 320 labrid species (50.7% of nominal species). We then employed stochastic character mapping to infer how and when cleaning behavior evolved. We estimate that cleaning has independently evolved 26-30 times in the Labridae, and all such events likely occurred no earlier than in the late Miocene. Given the current sampling and pattern of transitions, we hypothesize that the majority of facultative or obligate cleaning may have evolved through heterochrony.


Oesophageal bacterial biofilm changes in gastro-oesophageal reflux disease, Barrett's and oesophageal carcinoma: association or causality?
Barrett's oesophagus (BO) and gastro-oesophageal reflux disease (GERD) are precursors of oesophageal adenocarcinoma (OAC). There is an oesophageal biofilm, which changes in disease, but its role in aetiopathogenesis remains unclear. To define the oesophageal microbiota of patients with GERD, BO and OAC compared with controls and to investigate mucosal responses related to the microbiota. Cultural analysis identified the dominant bacterial species from a subset of each disease group. Based on this, molecular techniques were used to define the cohort. Host responses were analysed in tissues and co-culture experiments. A total of 111 species belonging to 26 genera were isolated. There was a significant decrease in bacterial counts in the GERD and BO groups for all genera except Campylobacter, which colonised GERD and Barrett's patients in increasing numbers. Campylobacter concisus was the dominant species. This relationship was not seen in the cancer group. Significant increases in IL-18 were seen in GERD and BO colonised by Campylobacter. This study defines differences in the oesophageal biofilm in disease states, revealing the emergence of C. concisus as the dominant new colonist in the refluxed oesophagus. We also associate the presence of these bacteria with increased expression of cytokines related to carcinogenesis.


Zebra leaf 15, a receptor-like protein kinase involved in moderate low temperature signaling pathway in rice.
Zebra leaf mutants are an important resource for studying leaf colour in rice. In most such mutants, the zebra leaf phenotype results from defective chloroplast biogenesis. The molecular mechanism by which zebra leaves develop remains unclear, so additional zebra-leaf mutants need to be identified. We isolated a novel rice zebra-leaf mutant, named zebra leaf 15 (z15), which showed transversely striped leaves with yellow-green or white-green sectors, in which chloroplast structure was disturbed. Transmission electron microscopy revealed that the structure of various organelles was impaired in yellow/white sectors. Z15, a single-copy gene in the rice genome, encodes a receptor-like protein kinase. Subcellular localization analysis indicates that Z15 and z15 are localized on the plasma membrane. The expression of Z15 is induced by moderate low temperature (18 °C). The mutation of Z15 influenced the expression of two downstream genes, OsWRKY71 and OsMYB4, that were responsive to moderate low temperature. The results show that Z15 plays a crucial role in the early stages of the response to moderate low temperature in rice. We identified a novel zebra-leaf mutant (z15) that impaired chloroplast structure in rice, LOC_Os05g12680, encoding a receptor-like protein kinase. Further study indiceted that Z15 plays a crucial role in the early stages of the response to moderate low temperature in rice.


Excess length of stay and economic consequences of adverse events in Dutch hospital patients.
To investigate the average and extrapolated excess length of stay and direct costs of adverse events (AEs) and preventable AEs in Dutch hospitals, and to evaluate patient characteristics associated with excess length of stay and costs. Data of a large retrospective patient record review study on AEs was used. A stratified sample of 20 Dutch hospitals was included. Excess length of stay and costs attributable to AEs and preventable AEs were calculated and extrapolated to a national estimate. The association between patient characteristics and excess length of stay (and costs thereof) attributable to AEs and preventable AEs was investigated through multilevel linear regression analyses. A total of 2975 patient records were included in the analysis, of which 325 experienced one or more AEs. Hospital patients experiencing an AE stayed 5.11 (95% CI 3.91-6.30) more days in hospital and cost €2600 (95% CI €1968-€3232) more compared to those without an AE. There was no significant difference in days and costs between preventable and non-preventable AEs. Extrapolated to a national level, AEs cost more than €300 million, which was 1.3% of the national hospital care budget. Patients with hospital-acquired infections had a statistically significant longer length of stay compared to the reference group (patients with AEs on the cardiovascular system). This study showed that AEs lead to substantial excess length of stay and increased costs. Special attention should be paid to patients with AEs due to an hospital-acquired infection.


Removal of tumour cells from bone marrow: an evaluation of the available techniques.
Autologous bone marrow transplantation has offered a way of increasing the dose of drugs and radiotherapy which can be used to treat patients with malignant disease in an attempt to eradicate tumour. Bone marrow is taken prior to treatment and returned to the patient to 'rescue' haemopoietic function after ablative therapy is completed. Bone marrow removed for autograft may be contaminated with tumour cells at the time of harvest, and it is undesirable to return these to patients even though there are little data available concerning the number of tumour cells necessary to reseed various malignancies. This review considers the various methods available for removing tumour cells from bone marrow destined for autologous transplantation, and evaluates their advantages and disadvantages.


Chromosomal polymorphism in the house mouse (Mus domesticus) of Greece and Yugoslavia.
A total of 88 wild mice from the Dalmatian coast of Yugoslavia (35 animals), and Peloponnesus (30 animals) and Thebes (23 animals) on mainland Greece were karyotyped. In all but five animals Robertsonian translations were found. Mice from the Dalmatian region were homozygous for translocations Rb(5.15), Rb(6.12), Rb(8.17), Rb(9.13), and Rb(10.14); they were homo- or heterozygous for the translocation Rb(1.11). Some of them lacked the Rb(1.11) translocation altogether so that the diploid numbers in the Yugoslavian mice were 2n = 28, 29, 30, or 40. The mice from the vicinity of Olympia in northwestern Peloponnesus were homozygous for eight Robertsonian translocations: Rb(1.3), Rb(2.5), Rb(4.6), Rb(8.12), Rb(9.16), Rb(10.14), Rb(11.17), and Rb(13.15). Their diploid chromosome number was therefore 2n = 24. Mice from the vicinity of Patras in northwest Peloponnesus carried all except the first three of these eight translocations; their chromosome number was 2n = 30. Finally, the mice from Thebes were homozygous for translocations Rb(2.15), Rb(4.14), Rb(5.12), and Rb(10.13). They were homo- or heterozygous for Rb(6.9), Rb(8.17), and Rb(1.11); some mice lacked the Tb(1.11) translocation altogether. The translocations Rb(6.9)40Tu and Rb(10.13)42Tu represent new arm combinations not found previously in any wild mouse population. The remaining translocations have previously been found in different Mediterranean countries, in Scotland and in southern Germany. The findings suggest that each translocation arose only once and that different translocations have come together in different populations to generate a unique karyotype characterizing this population.


Prospective analysis of association between statin use and breast cancer risk in the women's health initiative.
Statins are a class of cholesterol-lowering drugs that affect many intracellular pathways that may have implications for chemoprevention against cancer. Epidemiologic data on statins and breast cancer are conflicting. We analyzed updated data from the Women's Health Initiative (WHI) to assess the relationship between statins and breast cancer risk. The population included 154,587 postmenopausal women ages 50 to 79 years, with 7,430 pathologically confirmed cases of breast cancer identified over an average of 10.8 (SD, 3.3) years. Information on statins was collected at baseline and years one, three, six, and nine. Self- and interviewer-administered questionnaires were used to collect information on risk factors. Cox proportional hazards regression was used to calculate HRs with 95% confidence intervals (CI) to evaluate the relationship between statin use and cancer risk. Statistical tests were two-sided. Statins were used by 11,584 (7.5%) women at baseline. The annualized rate of breast cancer was 0.42% among statin users and 0.42% among nonusers. The multivariable adjusted HR of breast cancer for users versus nonusers was 0.94 (95% CI, 0.83-1.06). In the multivariable-adjusted, time-dependent model, the HR for simvastatin was 0.87 (95% CI, 0.71-1.07). There was no significant trend by overall duration of use (P value for trend 0.68). There was no effect of tumor stage, grade, or hormone receptor status. Overall, statins were not associated with breast cancer risk. Our study is one of the largest prospective observational studies on this topic, and substantially adds to the literature suggesting no relationship between statins and breast cancer risk.


Mycoplasma pneumoniae in children with acute and refractory asthma.
The presence of Mycoplasma pneumoniae has been associated with worsening asthma in children. Sensitive assays have been developed to detect M pneumoniae-derived community-acquired respiratory distress syndrome (CARDS) toxin. To identify the frequency and persistence of M pneumoniae detection in respiratory secretions of children with and without asthma and to evaluate antibody responses to M pneumoniae and the impact of M pneumoniae on biological markers, asthma control, and quality of life. We enrolled 143 pediatric patients (53 patients with acute asthma, 26 patients with refractory asthma, and 64 healthy controls; age range, 5-17 years) during a 20-month period with 2 to 5 follow-up visits. We detected M pneumoniae using CARDS toxin antigen capture and polymerase chain reaction and P1 adhesin polymerase chain reaction. Immune responses to M pneumoniae were determined by IgG and IgM levels directed against CARDS toxin and P1 adhesin. pH was measured in exhaled breath condensates, and asthma control and quality of life were assessed using the Asthma Control Test and Pediatric Asthma Quality of Life Questionnaire. M pneumoniae was detected in 64% of patients with acute asthma, 65% with refractory asthma, and 56% of healthy controls. Children with asthma had lower antibody levels to M pneumoniae compared with healthy controls. Exhaled breath condensate pHs and asthma control and quality of life scores were lower in M pneumoniae-positive patients with asthma. The results suggest that M pneumoniae detection is common in children, M pneumoniae detection is associated with worsening asthma, and children with asthma may have poor humoral immune responses to M pneumoniae.


Anthocyanin supplementation improves obesity-related inflammatory characteristics: A systematic review and meta-analysis of randomized controlled trials.
The relationship between anthocyanin intake and obesity-related inflammatory markers remains unclear in existing research. To investigate this, we hypothesized that anthocyanin supplementation could reduce plasma concentrations of inflammatory markers, including C-reactive protein (CRP), tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), vascular cell adhesion molecule-1, and other cytokines in obesity. We conducted a systematic search of PubMed, Web of Science, Scopus, SinoMed, and other related literature and identified 16 randomized controlled trials that met our inclusion criteria. Our findings showed that anthocyanin intake was significantly associated with a reduction in vascular cell adhesion molecule-1 mean plasma concentrations (-53.56 ng/mL; 95% confidence interval [CI], -82.10 to -25.03). We also observed a modest decrease in CRP (-0.27 ng/mL; 95% CI, -0.58 to 0.05), TNF-α (-0.20 ng/mL; 95% CI, -0.54 to 0.15), and IL-6 (-0.53 ng/mL; 95% CI, -1.16 to 0.10) mean plasma concentrations. Subgroup analysis revealed that anthocyanin intake tended to decrease CRP and IL-6 concentrations in overweight or dyslipidemic individuals. Additionally, the intervention duration subgroup analysis showed that anthocyanin supplementation had a stronger effect on plasma IL-6 and TNF-α in participants after 8 to 12 weeks of intervention. In conclusion, our meta-analysis indicated that anthocyanin supplementation can effectively reduce obesity-related inflammatory markers associated with chronic low-grade inflammation.


The automatic detection of known beta-propeller structural motifs from protein tertiary structure.
Following our previous work on the analysis of 'structural plasticity' associated with the beta-propeller structural motifs, we have now developed a simple method that can automatically detect all the known beta-propellers in protein tertiary structure, given a list of Protein Data Bank (PDB) codes as input to the computer program. Our beta-propeller detection (BPD) method identifies the location of beta-propellers in the protein structure, specifies the beta-propeller type, the beta-sheet associated beta-strand pattern and the structurally similar beta-propellers observed in other proteins. When tested on 21,566 proteins in the PDB, the BPD method was capable of correctly identifying all the known 245 beta-propellers described in the structural classification of proteins (SCOP) with the number of false positives detected being less than 0.2%. Forty-one false positives were detected that correspond to eight known protein families. When compared with some of the popular web-based programs that can automatically detect 'structural similarities' between the query and target proteins, our method has the advantage of also being capable of detecting beta-propellers associated with 'structural plasticity' and in situations where the target and query proteins differ in amino acid sequence length.


[The adaptational reactions in mild experimental craniocerebral trauma and its combination with alcoholic intoxication].
The article deals with the results of experimental study of adaptation reactions in mild craniocerebral trauma (MCCT) combined with acute alcoholic intoxication on the basis of appraisal of the dynamics of endocrine and neuromediator shifts. A stress character of these shifts was revealed. Acute alcoholic intoxication levelled the stress reaction occurring in MCCT.


Subjective symptoms contributing to the quality of life of rheumatoid arthritis patients with clinical remission from the IORRA database.
To explore patient-reported outcomes (PROs) related to quality of life (QOL) in patients with rheumatoid arthritis (RA) who achieved clinical remission. In the Institute of Rheumatology, Rheumatoid Arthritis dataset, RA patients >18 years old who met the simplified disease activity index (SDAI) remission criteria in April 2017 were enrolled in this analysis. Pain-visual analogue scale (pain-VAS) (0-100 mm), patient's global assessment of disease activity (Pt-GA; 0-100 mm), Japanese version of the Health Assessment Questionnaire, duration of morning joint stiffness, and fatigue [Checklist Individual Strength 8R (CIS)] were the tools used to evaluate PROs. To assess the contribution of each PRO to the European QOL-5 Dimensions-5 Level (EQ-5D-5L) score, an analysis of variance was conducted. Among the 2443 patients with remission, the mean EQ-5D-5L was 0.9. The mean pain-VAS and Pt-GA were 7.2 and 7.4, respectively. Factors that significantly contributed to the EQ-5D-5L were pain-VAS (48.8%), CIS score (18.1%), and Pt-GA (15.6%). Around 82.5% of the variance in EQ-5D-5L was explained by the three PROs. This study demonstrated that pain-VAS, CIS, and Pt-GA were significant contributors to the EQ-5D-5L score in patients with RA who achieved the simplified disease activity index remission criteria.


Correction to: Interdependence of laforin and malin proteins for their stability and functions could underlie the molecular basis of locus heterogeneity in Lafora disease.
Correction to: J. Biosci. 40(5), December 2015, 863-871 https://doi.org/10.1007/s12038-015-9570-0 The image of anti-Myc blot of figure 2C (third panel; Malin-Myc [C26S]) was inadvertently used once again for the c-tubulin loading control of figure 2B. The revised figure 2B with the correct image of the c-tubulin loading control is given below. The interpretation and conclusion provided in the article do not change because of the correction.


Influence of IFNgamma co-expression on the safety and antiviral efficacy of recombinant fowlpox virus HIV therapeutic vaccines following interruption of antiretroviral therapy.
Therapeutic immunization to stimulate host immune responses and control human immunodeficiency virus (HIV-1) replication is being investigated as a supplementary treatment for the management of HIV infection. On completion of an earlier study involving three vaccinations while taking combination antiretroviral therapy (CART), twenty-five subjects with plasma viral load (pVL) <50 copies/mL received a booster vaccination with either placebo (n = 7); fowl pox vaccine (rFPV) expressing HIV-1 Gag/Pol; [partial construct- PC (n = 8)] or rFPV coexpressing HIV-1 Gag/Pol and human interferon gamma[full construct - FC (n = 10)]. One week after the booster vaccination, participants stopped ART and were monitored for safety, pVL and immunological parameters for < or =20 weeks. The time weighted mean change (SD) from baseline plasma HIV RNA was 1.80 (0.72), 1.78 (0.91) and 0.96 (0.91) log(10) copies/mL for placebo, PC and FC recipients respectively (p = 0.06; mean differences between placebo and FC). Laboratory evaluations did not reveal differences in anti-HIV specific immune responses between study arms. No difference between treatment arms for host genetic factors known to affect pVL was demonstrated. In conclusion, vaccination with FC was associated with a trend toward lower rates of HIV replication following cessation of ART relative to placebo or PC. The promising antiretrovirological effect supports further study of FC in a larger trial with a broader population of patients with HIV disease.


Radioautographic demonstration of receptors for epidermal growth factor in various cells of the oral cavity.
Mouse iodinated epidermal growth factor (EGF) was localized by light and electron microscopic radioautography in basal cells of oral epithelium, papillary cells of the enamel organ, periodontal ligament fibroblasts, preodontoblast precursor cells, and preosteoblasts of the alveolar bone of 13-day-old Sprague-Dawley rats. The specificity of binding in these cells was suggested by an observed reduction of about 90% in the labeling when excess unlabeled EGF was injected along with the 125I-EGF. In contrast, fully differentiated cells, such as ameloblasts, odontoblasts, and osteoblasts, were only poorly labeled. Quantitative analysis of the light microscopic radioautographs revealed that the papillary cells had the highest level of labeling (5.5 grains per 100 micron 2 of cell area). The significance of the rather high labeling of the preosteoblasts of the alveolar bone and the fibroblasts of the periodontal ligament is unknown. However, the well-known effect of EGF in producing precocious eruption of teeth may be a consequence of an effect on these two cell types.


Mechanisms of nuclear protein import.
The past two years have seen a significant increase in our understanding of nuclear protein import. Five cytosolic import factors have been identified, two of which have been shown to directly interact with components of the nuclear pore complex. These findings enable refinement of previous models for steps in the nuclear import pathway, and provide a framework for future research.


Assessment of Kidney Injury as a Severity Criteria for Clostridioides Difficile Infection.
The Infectious Diseases Society of America (IDSA) and Society for Healthcare Epidemiology of America (SHEA) revised their Clostridioides difficile infection (CDI) severity classification criteria in 2017 to include an absolute serum creatinine (SCr) value above a threshold (≥1.5 mg/dL) rather than a relative increase from baseline (≥1.5 times the premorbid level). To date, how to best define kidney injury as a CDI disease severity marker has not been validated to assess severe outcomes associated with CDI. This multicenter cohort study included adult hospitalized patients with CDI. Patients were assessed for the presence of acute kidney injury (AKI), chronic kidney disease (CKD), and CDI severity using the 2010 and 2017 IDSA/SHEA CDI guidelines. Primary outcome was all-cause inpatient mortality. The final study cohort consisted of 770 CDI episodes from 705 unique patients aged 65 ± 17 years (female, 54%; CKD, 36.5%; AKI, 29.6%). Eighty-two episodes (10.6%) showed discordant severity classification results due to the inclusion of more patients with preexisting CKD in the severe disease category using an absolute SCr threshold criterion. The absolute SCr criterion better correlated with all-cause mortality (odds ratio [OR], 4.04; 95% confidence interval [CI], 1.76-9.28; P = .001) than the relative increase in SCr (OR, 1.34; 95% CI, 0.62-2.89; P = .46). This corresponded to an increased likelihood of the 2017 CDI severity classification criteria to predict mortality (OR, 5.33; 95% CI, 1.81-15.72; P = .002) compared with the 2010 criteria (OR, 2.71; 95% CI, 1.16-6.32; P = .02). Our findings support the 2017 IDSA/SHEA CDI severity classification criteria of a single pretreatment SCr in future CDI guideline updates.


Natural birth-induced UCP2 in brain development.
The prevalence of convenience-driven delivery via Caesarian-section (C-section) has been steadily increasing in Western as well as in developing societies. Today, significantly more deliveries occur via surgical means than medical necessity mandates. Despite tremendous advances in surgical techniques and pre- and post-operative surgical care, there remains to be limited understanding on the physiological influence of vaginal birth with particular emphasis on long term outcome on the offspring. Serendipitously, we recently uncovered in mice that vaginal birth induces significantly higher level of Ucp2 mRNA expression in the hippocampus than experimental C-section. We showed that during the early postnatal period, UCP2 expression promotes neuronal differentiation, axonal outgrowth and synapse formation in the hippocampus. We also observed that Ucp2 knockout animals show adult behaviors associated with the hippocampal formation that is different from wild type littermates. Taken together these observations suggest that vaginal birth may play a crucial role in determining adult brain structure and function that is different from that of the effect of surgically assisted birth.


Pharmacotherapy: Ketamine and Esketamine.
Ketamine and esketamine have rapid-onset antidepressant effects and may be considered for the management of treatment-resistant depression. Intranasal esketamine has regulatory approval in the United States and European Union. Intravenous ketamine is often administered off-label as an antidepressant, though no standard operating procedures exist. Repeated administrations and the use of a concurrent standard antidepressant may maintain antidepressant effects of ketamine/esketamine. Possible adverse effects of ketamine and esketamine include psychiatric, cardiovascular, neurologic and genitourinary effects, and the potential for abuse. The long-term safety and efficacy of ketamine/esketamine as antidepressants require further study.


Evaluation of a "Subintern" Role: Action Over Observation For Final-Year Medical Students in Surgery.
This study sought to investigate the potential of a subinternship apprentice role as a structured experiential learning opportunity by enabling final-year medical students to learn through action rather than observation. A convenience sample of 65 final-year medical students was allocated to "intern shadow" (control group) or "subintern" (research group) for 2 weeks during the first clinical attachment of their final year. "Intern shadow" involved direct observation of the intern role, whereas "subintern" involved a fully immersive performance of the intern role under direct supervision by the surgical team. All students completed an evaluation form that was devised in-house and based on course learning objectives. This consisted of Likert scales and free text. Wilcoxon rank sum test results showed that students participating in the "subintern" group had a statistically significantly better clinical experience than the "intern-shadow" group regarding role modeling (p = 0.0002), learning and teaching (p = 0.0021), organization and support (p = 0.0219), and patient interaction (p = 0.0063). The "subintern" apprentice role shows strong potential for enabling final-year students to learn on the job by actually performing the intern role rather than merely observing an intern at work. Subinterns reported feeling valued within the team structure and expressed a desire to emulate expert practice directly experienced through close collaboration with consultant surgeons.


Chromomycosis caused by Philaphora pedrosoi in eastern Nigeria.
Three cases of chromomycosis from Eastern Nigeria are described. The aetiological agent was identified as Phialophora pedrosoi. Amphotericin B cleared most of the lesions in one case while 5-fluorocytosine gave a complete cure in another case.


Substance P receptor desensitization in the dorsal horn: possible involvement of receptor-G protein complexes.
The repeated administration of a high dose of substance P (SP) onto the spinal cord has been shown to attenuate behavioral responses to an intense heat (tail-flick) or noxious mechanical stimulus (paw pressure). Studies performed to investigate the action of spinal SP have suggested that changes in behavioral responses involve endogenous opiate or neurokinin systems. This study was performed to investigate whether the binding characteristics of SP receptors in the dorsal horn are altered following successive administration of SP. Two populations of [3H]-SP binding sites were distinguished on the basis of their binding affinity. Gpp(NH)p, a stable analogue of GTP, decreased the size and affinity of the high affinity binding component selectively labelled with [125I]-Bolton Hunter-SP. Repeated intrathecal administration of SP (15 micrograms) which reduced behaviors also reduced the number and affinity of high affinity binding sites. Thus, attenuated behaviors in response to repeated administration of SP are paralleled by an alteration of SP binding in the dorsal horn. The altered agonist affinity seen under desensitizing conditions raises the possibility that SP receptor desensitization involves an uncoupling of receptor-G protein complexes.


KiSS-1 and GPR54 as new players in gonadotropin regulation and puberty.
The recent identification of loss-of-function mutations in the gene encoding GPR54, the receptor for the KiSS-1-derived peptides, kisspeptins, has highlighted a previously unrecognized pathway in the physiologic regulation of puberty and reproduction. Patients with loss-of-function mutations in GPR54 have idiopathic hypogonadotropic hypogonadism, and mice lacking GPR54 similarly fail to undergo puberty and have immature reproductive organs and low levels of sex steroids and gonadotropins. These observations have led to the hypothesis that kisspeptins activate hypothalamic GnRH release, thereby serving as a pivotal factor in the pubertal activation of the reproductive cascade. This hypothesis is supported by subsequent studies in rodent and primate models that have demonstrated localization of KiSS-1 mRNA in the hypothalamus, colocalization of GPR54 in GnRH neurons, GnRH-dependent activation of LH and FSH release by intracerebroventricular or peripheral administration of kisspeptin, and increased hypothalamic KiSS-1 and GPR54 mRNA levels at the onset of puberty. Taken together, these findings weave a compelling case for a role of the kisspeptin-GPR54 system in the activation of GnRH neurons at the time of pubertal awakening of the reproductive axis.


Functional profiling of LDLR variants: Important evidence for variant classification: Functional profiling of LDLR variants.
Familial Hypercholesterolemia (FH) is a semidominant disorder of the lipid metabolism associated with premature atherosclerosis and coronary heart disease. So far, about 3,000 unique LDLR variants have been described, most of which lack functional evidence proving their effect on LDLR function, despite the important role that functional studies play in variant classification. In this work, we aimed to functionally characterize 13 rare missense variants, identified worldwide and in Portugal, in clinical FH patients. LDLR-deficient CHO-ldlA7 cells were transfected with plasmids carrying different LDLR variants generated by site-directed mutagenesis. LDLR activity and expression were assessed by FACS. 11/13 variants affect LDLR function (p.Cys109Phe; p.Cys143Arg; p.Glu267Lys; p.Cys352Ser; p.Ile451Thr; p.His485Gln; p.Asp492Asn; p.Val500Ala; p.Gly529Arg; p.Phe614Ile; p.Glu626Lys) and 2/13 are inconclusive (p.Arg81Cys; p.Gly98Arg;). Of the 13 variants studied, 8 were classified as VUS by ACMG criteria, but for 7 of these 8, our functional studies were able to reassign them as Likely pathogenic or Pathogenic. For an accurate diagnosis, an effort must be made to improve functional characterization of putative disease-causing variants.


Influence of category identity on letter matching: conceptual penetration of visual processing or response competition?
Participants performed same-different matching tasks, with physical-identity instructions, on letter pairs composed from the letters B, b, and p. The letters in a pair were presented simultaneously or sequentially, with the experiments differing in whether (1) the letters could appear in two or four positions, (2) two or five SOAs were used, (3) the font was Arial or one in which the two loops of the letter B were of the same size, and (4) p did or did not occur in same pairs. With sequential presentation, different RTs were longer when the letters had the same name (Bb; within-category pair) than when they did not (Bp; between-category pair), replicating a finding by Lupyan et al. (2010). However, unlike in their study, this category effect was also significant with simultaneous presentation, tending to be nonsignificantly smaller for RTs but larger for accuracy than that obtained with sequential presentation. A similar pattern was observed when we removed a bias to respond different whenever the letter p was detected in the experiments in which p did not appear in same pairs. The presence of a category effect with simultaneous presentation is predicted by a response competition account, but not by Lupyan et al.'s conceptual-penetration-of-visual-processing account.


Obese and nonobese patients with essential hypertension show similar N-terminal proBNP plasma levels.
Multiple studies have focused on the influence of obesity on natriuretic peptide levels. However, the effect of obesity on amino-terminal propeptide of B-type natriuretic peptide (NT-proBNP) levels in hypertensive (HT) patients remains uncertain. We studied 252 asymptomatic patients (60 +/- 13 years, 136 men) with essential HT. A routine physical examination, anthropometry, laboratory analyses, echo-Doppler study, and NT-proBNP level determination were performed. NT-proBNP levels were similar in both obese and nonobese HT (median 56 (25-130) pg/ml vs. median 51 (26-129) pg/ml, P = 0.488). No significant differences were found in obese or nonobese patients with left ventricular hypertrophy (LVH) (median 135 (73-425) pg/ml vs. median 151 (64-274) pg/ml, P = 0.597). The area under the curve was 0.89 +/- 0.03 for NT-proBNP to diagnose LVH in the obese HT patients and 0.88 +/- 0.03 in the nonobese. A logistic regression analysis showed that age, gender, and left ventricular mass index (LVMI) were independent predictors of NT-proBNP levels. Body mass index (BMI) was not significantly associated with NT-proBNP in LVH HT patients. Obesity is not statistically associated with NT-proBNP levels in HT asymptomatic patients. The same results were observed in our group of patients with LVH. These data are in contrast with those previously found in heart failure, and raise questions about the role of obesity per se as primary cause of decreased NT-proBNP levels in other pathophysiological conditions.


Iron-binding fragments from the N-terminal and C-terminal regions of human lactoferrin.
Digestion of lactoferrin with pepsin at pH3.0 gave an iron-binding half-molecule that represents the C-terminal part of the native protein. Tryptic or chymotryptic digestion of 30%-iron-saturated lactoferrin yielded the N- and C-terminal half molecules, which could be separated by DEAE-Sephadex chromatography. The N- and C-terminal fragments did not show any immunological cross-reaction. The carbohydrate of lactoferrin was distributed equally between the two fragments.


Acid-sensing ion channels are expressed in the ventrolateral medulla and contribute to central chemoreception.
The role of acid-sensing ion channels (ASICs) in the ventrolateral medulla (VLM) remains uncertain. Here, we found that ASIC1a and ASIC2 are widely expressed in rat medulla, and the expression level is higher at neonatal stage as compared to adult stage. The two ASIC subunits co-localized in medualla neurons. Furthermore, pH reduction triggered typical ASIC-type currents in the medulla, including the VLM. These currents showed a pH50 value of 6.6 and were blocked by amiloride. Based on their sensitivity to psalmotoxin 1 (PcTx1) and zinc, homomeric ASIC1a and heteromeric ASIC1a/2 channels were likely responsible for acid-mediated currents in the mouse medulla. ASIC currents triggered by pH 5 disappeared in the VLM neurons from ASIC1-/-, but not ASIC2-/- mice. Activation of ASICs in the medulla also triggered neuronal excitation. Moreover, microinjection of artificial cerebrospinal fluid at a pH of 6.5 into the VLM increased integrated phrenic nerve discharge, inspiratory time and respiratory drive in rats. Both amiloride and PcTx1 inhibited the acid-induced stimulating effect on respiration. Collectively, our data suggest that ASICs are highly expressed in the medulla including the VLM, and activation of ASICs in the VLM contributes to central chemoreception.


A concordance study of the Altona RealStar Varicella-Zoster virus real-time quantitative PCR and in-house conventional qualitative PCR.
This study compared the Altona RealStar™ VZV Kit 1.0 real time quantitative VZV PCR with in-house qualitative conventional VZV PCR on cerebrospinal fluid, mucocutaneous, and other uncommon clinical specimens. Overall, positive and negative agreement percentages were respectively 97.9% (95%CI: 93.8-99.6), 100.0% (95%CI: 93.1-100.0), and 96.3% (95%CI: 89.4-99.2) while Cohen's kappa statistic value was 0.96 (95%CI: 0.91-1.00). RealStar™ VZV quantitative PCR assay reported average quantitative viral loads of 4.4 × 105 and 1.1 × 107 copies/mL in cerebrospinal fluid and cutaneous specimens, respectively (P < 0.01). RealStar™ VZV PCR assay showed excellent agreement with in house conventional assay for various clinical specimens.


Elective rotations and their impact on resident and faculty decision making.
Restrictions on resident-duty work hours have led to fewer opportunities to participate in elective rotations. To investigate the potential impact of elective rotations on resident and faculty physicians using allergy/immunology (A/I) as a model. A questionnaire was sent to 375 primary care physicians. It assessed demographic information, training background, A/I-referral history and intentions, and perceived knowledge of A/I-specific issues. A total of 228 surveys were returned. Resident physicians who were more senior in their training or who had taken an A/I rotation were more likely to have referred a patient to A/I in the past, more likely to refer a patient to A/I for chronic sinusitis, and more likely to report a greater knowledge of A/I topics. Faculty physicians with a history of an A/I rotation were more likely to feel knowledgeable about A/I topics. Elective rotations are positively correlated with perceived knowledge, referral intentions, and past history of referrals.


Homogeneous-heterogeneous interfaces in 2D/2D CoAl/Co9S8/Ni3S4 heterostructures for electromagnetic wave absorption.
Exploring electromagnetic wave (EMW) absorbers with ultrathin matching thickness (d ≤ 1.5 mm), strong reflection loss (RL ≤ -50 dB), and wide effective absorption bandwidth (EAB, RL ≤ -10 dB) is urgent and essential for reducing EMW radiation and interference. Herein, a 2D/2D CoAl/Co9S8/Ni3S4 heterostructure was constructured using simple hydrothermal and pyrolysis methods. 2D porous CoAl nanosheets and 2D Co9S8/Ni3S4 ultrathin nanosheets are assembled by small nanoparticle chains. Strikingly, the CoAl/Co9S8/Ni3S4 heterostructure exhibits remarkable EMW absorption performance with a RL value of -61.56 dB, a high EAB of 4 GHz, and an ultrathin matching thickness of 1.25 mm. Mechanism investigations reveal that the CoAl/Co9S8/Ni3S4 heterostructure delivers dual metal sulfides behavior, high specific surface area, strong interactions, rich defects (N doping), and abundant homogeneous and heterogeneous interfaces, which promote good impedance matching, dielectric loss (interface polarization, conductive loss, and dipole polarization), as well as magnetic loss (natural resonance, exchange resonance, and eddy current loss) characteristics. This work can provide insights into the mechanism of dual metal sulfides used as high-performance EMW absorbers and deepen our understanding of the design and application of 2D/2D heterostructures.


Depression in people with epilepsy in West China: Status, risk factors and treatment gap.
To evaluate the prevalence, suicidality and treatment gap (TG) of depression in people with epilepsy (PWE) in West China, and to explore the related risk factors. A consecutive cohort of PWE from West China Hospital was recruited. The Chinese version of the Neurological Disorders Depression Inventory for Epilepsy scale was used to assess depression and suicidality. Prevalence and TG of depression were calculated. Logistic regression analysis was used to assess risk factors of depression and suicidality. Among a total of 461 participants, there were 29.9% with depression and 15.8% had suicidality. Being female, an unmarried status, disease course, seizure frequency, seizure occurrence in the last 6 months, and focal impaired awareness seizure were risk factors for depression. Depression was an independent risk factor for suicidality. The TG of depression was 72.5%. The TG was larger in the 30-40 year age group, in rural populations, and among those with a disease course between 5 and 10 years. The TG figures dropped with increasing annual family income. The prevalence of depression and the rate of suicide risk for PWE in West China are high. A large TG for depression exists in PWE. Clinicians should focus on depression and suicidality for PWE and identify the related risk factors in a timely manner so as to reduce the incidence of depression and suicide. Management including public education as well as professional training of neurologists are necessary to fill the TG.


Distribution of substance P, somatostatin and neurotensin in the human hypothalamus.
The distribution of substance P, somatostatin and neurotensin in the human hypothalamus was studied by radioimmunoassay analysis of 11 nuclei and areas within the hypothalamus, in 10 human post-mortem cases. The samples were microdissected according to a recently developed standardized method. Substance P was present in higher concentrations both rostrally in the hypothalamus, and ventrally. Somatostatin was found predominantly in the basal tuberal region, and neurotensin in the anterior hypothalamic area. The chromatographic behaviour of human hypothalamic neurotensin is also presented.


Mavacamten in Patients With Hypertrophic Cardiomyopathy Referred for Septal Reduction: Week 56 Results From the VALOR-HCM Randomized Clinical Trial.
There is an unmet need for novel medical therapies before recommending invasive therapies for patients with severely symptomatic obstructive hypertrophic cardiomyopathy (HCM). Mavacamten has been shown to improve left ventricular outflow tract (LVOT) gradient and symptoms and may thus reduce the short-term need for septal reduction therapy (SRT). To examine the cumulative longer-term effect of mavacamten on the need for SRT through week 56. This was a double-blind, placebo-controlled, multicenter, randomized clinical trial with placebo crossover at 16 weeks, conducted from July 2020 to November 2022. Participants were recruited from 19 US HCM centers. Included in the trial were patients with obstructive HCM (New York Heart Association class III/IV) referred for SRT. Study data were analyzed April to August 2023. Patients initially assigned to mavacamten at baseline continued the drug for 56 weeks, and patients taking placebo crossed over to mavacamten from week 16 to week 56 (40-week exposure). Dose titrations were performed using echocardiographic LVOT gradient and LV ejection fraction (LVEF) measurements. Proportion of patients undergoing SRT, remaining guideline eligible or unevaluable SRT status at week 56. Of 112 patients with highly symptomatic obstructive HCM, 108 (mean [SD] age, 60.3 [12.5] years; 54 male [50.0%]) qualified for the week 56 evaluation. At week 56, 5 of 56 patients (8.9%) in the original mavacamten group (3 underwent SRT, 1 was SRT eligible, and 1 was not SRT evaluable) and 10 of 52 patients (19.2%) in the placebo crossover group (3 underwent SRT, 4 were SRT eligible, and 3 were not SRT evaluable) met the composite end point. A total of 96 of 108 patients (89%) continued mavacamten long term. Between the mavacamten and placebo-to-mavacamten groups, respectively, after 56 weeks, there was a sustained reduction in resting (mean difference, -34.0 mm Hg; 95% CI, -43.5 to -24.5 mm Hg and -33.2 mm Hg; 95% CI, -41.9 to -24.5 mm Hg) and Valsalva (mean difference, -45.6 mm Hg; 95% CI, -56.5 to -34.6 mm Hg and -54.6 mm Hg; 95% CI, -66.0 to -43.3 mm Hg) LVOT gradients. Similarly, there was an improvement in NYHA class of 1 or higher in 51 of 55 patients (93%) in the original mavacamten group and in 37 of 51 patients (73%) in the placebo crossover group. Overall, 12 of 108 patients (11.1%; 95% CI, 5.87%-18.60%), which represents 7 of 56 patients (12.5%) in the original mavacamten group and 5 of 52 patients (9.6%) in the placebo crossover group, had an LVEF less than 50% (2 with LVEF ≤30%, one of whom died), and 9 of 12 patients (75%) continued treatment. Results of this randomized clinical trial showed that in patients with symptomatic obstructive HCM, mavacamten reduced the need for SRT at week 56, with sustained improvements in LVOT gradients and symptoms. Although this represents a useful therapeutic option, given the potential risk of LV systolic dysfunction, there is a continued need for close monitoring. ClinicalTrials.gov Identifier: NCT04349072.


Toxicity of an herbicide and adjuvant to saltmarsh invertebrates in the management of invasive grass; Comparative laboratory and field tests.
Coastal weeds are often treated with herbicides without knowledge of non-target impacts, and toxicity data from standardized test species can have limited applicability. We evaluated toxicity to invertebrates from Fusilade Forte® and the adjuvant Hasten® in the control of invasive salt marsh grass, Spartina anglica. For 3 of 4 local invertebrates, Fusilade Forte® was moderately toxic (96h LC50 5.4-144mgL(-1)), whereas Hasten® was less toxic (14.2-450mgL(-1)). For most species, the mixture was more toxic than the herbicide alone, with 96h LC50 reduced 23-45%. However, a field experiment applying typical concentrations (1000×the lowest 96h LC50) showed low concentrations of herbicide residues and no detrimental impacts on invertebrates over 6months. The results reveal the importance of testing locally relevant species for potential toxicity, and of comparison tests with field exposures to determine the realised toxicity in nature.


The effect of work motivation on a sample of nurses in an Italian healthcare setting.
Research in Human Resources Management has highlighted that implementing strategies for increasing employees' motivation can be an advantage for organizations because this improves their performance. Self Determination Theory (SDT) was used to analyze the mediating role of autonomous vs. controlled motivation on the relationship between organizational factors (perceived organizational support and job autonomy) and work outcomes (affective commitment, job satisfaction, and intent to quit). A total of 304 nurses from an Italian hospital participated in the study. The results supported the importance of the mediating role of autonomous motivation on the relationship between both perceived organizational support (POS) and job autonomy, and work outcomes. Conversely, controlled motivation did not show mediating effects on that relationship. POS was directly related to affective commitment and job satisfaction. Practical implications for organizations and employees are discussed along with a call for further research in this area.


The other side of the coin: mesenchymal stromal cell immortalization beyond evasion of senescence.
Mesenchymal stromal cells (MSC) are promising options to cellular therapy to several clinical disorders, mainly because of its ability to immunomodulate and differentiate into different cell types. Even though MSC can be isolated from different sources, a major challenge to understanding the biological effects is that the primary cells undergo replicative senescence after a limited number of cell divisions in culture, requiring time-consuming and technically challenging approaches to get a sufficient cell number for clinical applications. Therefore, a new isolation, characterization, and expansion is necessary every time, which increases the variability and is time-consuming. Immortalization is a strategy that can overcome these challenges. Therefore, here, we review the different methodologies available to cellular immortalization, and discuss the literature regarding MSC immortalization and the broader biological consequences that extend beyond the mere increase in proliferation potential.


Utilization of vinasse for the production of polyhydroxybutyrate by Haloarcula marismortui.
Vinasse, a recalcitrant waste of the ethanol industry was employed for the production of polyhydroxyalkanoate (PHA) by the extremely halophilic archaeon, Haloarcula marismortui in shake flasks. The PHA was recovered by osmotic lysis of the cells and subsequent purification by sodium hypochlorite and organic solvents. Through UV-vis spectroscopy, differential scanning calorimetry, Fourier transform infrared, and nuclear magnetic resonance spectroscopy, the PHA was found to have characteristics very similar to that of the standard polyhydroxybutyrate (PHB) from Sigma. Inhibitory effect of polyphenols contained in vinasse was assessed by a quick and reliable cup-plate agar-diffusion method. Raw vinasse (10%) was utilized leading to accumulation of 23% PHA (of cell dry weight) and following an efficacious pre-treatment process through adsorption on activated carbon, 100% pre-treated vinasse could be utilized leading to 30% accumulation of PHB by H. marismortui. Maximum specific growth rate, specific production rate, and volumetric productivity attained using 10% raw vinasse were comparable to that obtained using a previously reported nutrient deficient medium (NDM), while the values with 100% pre-treated vinasse were higher than that determined using NDM medium. This is the first report of polyhydroxybutyrate production by a halophilic microorganism utilizing vinasse.


Inflammation and infections in asthma.
Asthma is driven by an inflammatory response against normally harmless environmental inorganic and organic compounds in the respiratory tract. Immune responses to airborne pathogens such as viruses and bacteria may reduce the allergic responses but are also known to trigger asthma attacks and eventually lead to severe disease condition. To investigate the role of respiratory pathogens concerning the induction or protection against acute or chronic asthma manifestations. We included 131 articles for the final review according to their relevance with the subject. There is apparently contradictory interaction of respiratory germs in the airways of asthmatics which may be protective on one angle but deleterious on the other. The relationship between inflammation and remodeling and the pathogenic role of viral and bacterial infection in the airways of asthmatic patients is still highly debatable and incompletely understood.


Synergism between environmental lighting and taurine depletion in causing photoreceptor cell degeneration.
Experiments were conducted to examine the possible interaction between retinal taurine depletion and environmental lighting in causing photoreceptor cell degeneration. Albino rats were raised from birth in either dim (2 lx) or relatively bright (300 lx) cyclic light. Beginning at weaning, half the animals from both light environments were taurine-depleted by treating them for 10 weeks with guanidinoethyl sulfonate as a 1% solution in their drinking water. The remaining animals were given ordinary tap-water and served as controls. In the 2 lx light environment, taurine depletion caused a decrease in electroretinogram (ERG) a- and b-wave amplitude of 36 and 46%, respectively; however, no photoreceptor cells were lost in this group. Tap-water controls kept in the 300 lx light environment had a 59- and 43% decrease in ERG a- and b-wave amplitude, respectively, and a 21% reduction in the number of photoreceptor cells. In contrast to the other groups, animals that were taurine-depleted in the 300 lx environment showed a marked retinal degeneration. ERG a- and b-wave amplitude was decreased by 94- and 89% respectively and there was a 62% loss of photoreceptor cells. The greatest cell loss occurred in the central superior region of the retina, in which the outer nuclear layer was typically reduced to one to two rows of nuclei. The results of a two-way analysis of variance applied to the data indicated that the effects of taurine depletion on the retina were greater in the 300 lx as compared with the 2 lx environment in terms of loss of photoreceptor cells and reduction in log ERG a- and b-wave amplitude. These findings demonstrate a synergism between environmental lighting and taurine depletion in causing photoreceptor cell degeneration.


Contextual Inferences, Nonlocality, and the Incompleteness of Quantum Mechanics.
It is known that "quantum non locality", leading to the violation of Bell's inequality and more generally of classical local realism, can be attributed to the conjunction of two properties, which we call here elementary locality and predictive completeness. Taking this point of view, we show again that quantum mechanics violates predictive completeness, allowing the making of contextual inferences, which can, in turn, explain why quantum non locality does not contradict relativistic causality. An important question remains: if the usual quantum state ψ is predictively incomplete, how do we complete it? We give here a set of new arguments to show that ψ should be completed indeed, not by looking for any "hidden variables", but rather by specifying the measurement context, which is required to define actual probabilities over a set of mutually exclusive physical events.


Choice of first-line antiretroviral therapy regimen and treatment outcomes for HIV in a middle income compared to a high income country: a cohort study.
The range of combination antiretroviral therapy (cART) regimens available in many middle-income countries differs from those suggested in international HIV treatment guidelines. We compared first-line cART regimens, timing of initiation and treatment outcomes in a middle income setting (HIV Centre, Belgrade, Serbia - HCB) with a high-income country (Royal Free London Hospital, UK - RFH). All antiretroviral-naïve HIV-positive individuals from HCB and RFH starting cART between 2003 and 2012 were included. 12-month viral load and CD4 count responses were compared, considering the first available measurement 12-24 months post-cART. The percentage that had made an antiretroviral switch for any reason, or for toxicity and the percentage that had died by 36 months (the latest time at which sufficient numbers remained under follow-up) were investigated using standard survival methods. 361/597 (61 %) of individuals initiating cART at HCB had a prior AIDS diagnosis, compared to 337/1763 (19 %) at RFH. Median pre-ART CD4 counts were 177 and 238 cells/mm(3) respectively (p < 0.0001). The most frequently prescribed antiretrovirals were zidovudine with lamivudine (149; 25 %) and efavirenz [329, 55 %] at HCB and emtricitabine with tenofovir (899; 51 %) and efavirenz [681, 39 %] at RFH. At HCB, a median of 2 CD4 count measurements in the first year of cART were taken, compared to 5 at RFH (p < 0.0001). Median (IQR) CD4 cell increase after 12 months was +211 (+86, +359) and +212 (+105, +318) respectively. 287 (48 %) individuals from HCB and 1452 (82 %) from RFH had an available viral load measurement, of which 271 (94 %) and 1280 (88 %) were <400 copies/mL (p < 0.0001). After 36 months, comparable percentages had made at least one antiretroviral switch (77 % HCB vs. 78 % RFH; p = 0.23). However, switches for toxicity/patient choice were more common at RFH. After 12 and 36 months of cART 3 % and 8 % of individuals died at HCB, versus 2 % and 4 % at RFH (p < 0.0001). In middle-income countries, cART is usually started at an advanced stage of HIV disease, resulting in higher mortality rates than in high income countries, supporting improved testing campaigns for early detection of HIV infection and early introduction of newer cART regimens.


Polycystic Kidney Disease-Related Disease Burden in Adolescents With Autosomal Dominant Polycystic Kidney Disease: An International Qualitative Study.
Little is known about symptoms and disease impacts in adolescents with autosomal dominant polycystic kidney disease (ADPKD). The objective of the study was to explore these issues from the adolescent patient's perspective. Observational, qualitative study. Eligible participants were 12-17 years old and had a diagnosis of ADPKD. Semi-structured interviews were conducted in 18 cities in 13 countries to elicit participant experiences of ADPKD-related symptoms and physical, social, and emotional impacts. Interviews were recorded, transcribed, and coded. Symptom and impact frequencies from the interviews were calculated, and representative quotes concerning elicited concepts were collated. Thirty-three participants (mean age, 14.6 years; 42.4% female) completed interviews. Frequently reported symptoms included urinary urgency (n = 10; 30.3%) and back pain (n = 9; 27.3%). Consistent with previous findings in adults, participants experienced 3 primary types of pain: dull kidney pain, severe or sharp kidney pain, and a feeling of fullness and/or discomfort. Reported disease impacts included avoiding sports and physical activity (n = 10; 30.3%), missing school (n = 6; 18.2%) and social activities (n = 6; 18.2%), and feeling worried (n = 6; 18.2%), sad (n = 4; 12.1%), or frustrated (n = 3; 9.1%) about the disease and their future. Approximately one-fifth of participants (n = 7; 21.2%) reported that they were bothered or impacted by dietary limitations (primarily the need for reduced sodium intake and increased water intake). The study had a small sample size. The researchers were unable to conduct focus groups with participants because of parental preferences. The findings from this exploratory study indicate that a substantial proportion of adolescents with ADPKD experience physical, social, and emotional impacts from their disease.


The effect of cognitive behavioral therapy for insomnia in schizophrenia patients with sleep Disturbance: A non-randomized, assessor-blind trial.
This non-randomized, assessor blind study evaluated the effects of cognitive behavioral therapy for insomnia (CBT-I) delivered in a group format on insomnia symptoms as well as psychotic, depressive, and anxiety symptoms in schizophrenia patients (n = 63) recruited from residential or rehabilitative facilities in Seoul, South Korea. Thirty-one patients received four sessions of CBT-I in groups of 2-9 patients in addition to usual care, while the control group (n = 32) received no additional intervention. The Insomnia Severity Index (ISI) and Pittsburgh Sleep Quality Index (PSQI), Psychotic Symptoms Rating Scale (PSYRATS), Anxiety Sensitivity Index (ASI), and Beck Depression Inventory (BDI) were administered at baseline, week 4, and week 8. Both groups showed significant time-group interactions on the ISI and PSQI. Post hoc testing showed that, compared to the control group, the CBT-I group showed significant reductions in ISI and PSQI at both week 4 and week 8. For the PSYRATS, ASI, and BDI scores, the CBT-I and control groups showed significant time-group interactions, but post hoc testing revealed no significant group differences at either week 4 or week 8. Therefore, CBT-I was effective for reducing insomnia symptoms in patients with schizophrenia and the effect lasted for 4 weeks after the intervention.


Crystal forms in pharmaceutical applications: olanzapine, a gift to crystal chemistry that keeps on giving.
This contribution reviews the efforts of many scientists around the world to discover and structurally characterize olanzapine crystal forms, clearing up inconsistencies in the scientific and patent literature and highlighting the challenges in identifying new forms amidst 60+ known polymorphs and solvates. Owing to its remarkable solid-state chemistry, olanzapine has emerged over the last three decades as a popular tool compound for developing new experimental and computational methods for enhanced molecular level understanding of solid-state structure, form diversity and crystallization outcomes. This article highlights the role of olanzapine in advancing the fundamental understanding of crystal forms, interactions within crystal structures, and growth units in molecular crystallization, as well as influencing the way in which drugs are developed today.


The effect of manganese and iron on mediating resuscitation of lactic acid-injured Escherichia coli.
Lactic acid can induce sublethal injury of E. coli through oxidative stress. In this study, we investigated changes in SOD activity, CAT activity, GSH production and ROS production during sublethal injury and resuscitation of E. coli. Then, the effect of manganese and iron during resuscitation were studied. Both cations (≥1 mmol l-1 ) significantly promoted the resuscitation of sublethally injured E. coli induced by lactic acid and shortened the repair time (P < 0·05). Conversely, addition of N,N,N',N'-tetrakis (2-pyridylmethyl) which is a metal chelator extended the repair time. Compared with minA, manganese and iron significantly improved SOD activity at 40, 80 and 120 min and decreased ROS production at 40 and 80 min, thereby recovering injured E. coli quickly (P < 0·05). The deletion of sodA encoding Mn-SOD, sodB encoding Fe-SOD or gshA/gshB encoding GSH significantly strengthened sublethal injury and extended the repair time (P < 0·05). It meant these genes-related oxidative stress played important roles in the acid resistance of E. coli and recovery of sublethal injury. Therefore, manganese and iron can promote the recovery of lactic-injured E. coli by the way of increasing SOD activity, scavenging ROS, and relieving oxidative stress.


Subarachnoid-pleural fistula after resection of a pancoast tumor with hyponatremia.
Resection of superior sulcus neoplasms is associated with a number of complications resulting from the extensive nature of the resection and the necessity to sacrifice certain adjacent structures. One of the complications of resection is the development of subarachnoid-pleural fistula, with the subsequent appearance of air in the cerebrospinal fluid circulation. We report a case in which a subarachnoid-pleural fistula led to persistent pneumocephaly in a patient who exhibited postoperative hyponatremia, confusion, and gait disturbance.


Synergistic upregulation of ADAMTS4 (aggrecanase-1) by cytokines and its suppression in knee osteoarthritic synovial fibroblasts.
The ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) family includes nine members with aggrecan-degrading activity, i.e., ADAMTS1, 4, 5, 8, 9, 15, 16, 18, and 20. However, their systematic expression profile in knee osteoarthritis (OA) synovium and effects of cytokines and growth factors on the expression in OA synovial fibroblasts remain elusive. In this study, expression of all nine aggrecanolytic ADAMTS species was assessed by quantitative real-time PCR in OA and control normal synovial tissues. OA synovial fibroblasts were treated with interleukin-1α (IL-1α), IL-1β, tumor necrosis factor-α (TNF-α), transforming growth factor-β (TGF-β), vascular endothelial growth factor165, and heparin-binding epidermal growth factor, and analyzed for the expression of the ADAMTS species. The signaling pathways and inhibition of ADAMTS4 expression by high-molecular-weight hyaluronan, adalimumab, tocilizumab, and signaling molecule inhibitors were studied. ADAMTS1, 4, 5, 9, and 16 were expressed in OA synovium, but only ADAMTS4 expression was significantly higher in OA as compared to normal synovium. IL-1α, TNF-α, and TGF-β markedly increased ADAMTS4 expression, while their effects were minimal for the other ADAMTS species. ADAMTS4 was synergistically upregulated by treatment with IL-1α and TNF-α, IL-1α and TGF-β, or IL-1α, TNF-α and TGF-β. The signaling molecules' inhibitors demonstrated that IL-1α-induced ADAMTS4 expression is predominantly through TGF-β-associated kinase 1 (TAK1), and the TNF-α-stimulated expression is via TAK1 and nuclear factor-κB (NF-κB). The TGF-β-promoted expression was through the activin receptor-like kinase 5 (ALK5)/Smad2/3, TAK1, and non-TAK1 pathways. Adalimumab blocked TNF-α-stimulated expression. ADAMTS4 expression co-stimulated with IL-1α, TNF-α and TGF-β was abolished by treatment with adalimumab, TAK1 inhibitor, and ALK5/Smad2/3 inhibitor. These data demonstrate marked and synergistic upregulation of ADAMTS4 by IL-1α, TNF-α and TGF-β in OA synovial fibroblasts, and suggest that concurrent therapy with an anti-TNF-α drug and inhibitor(s) may be useful for prevention against aggrecan degradation in OA.


Peer-Mentoring: A Call to Create Inclusive Programs to Support All Students' Ability to Be Successful.
Support for diverse nursing students needs to be built into existing peer-mentoring programs to foster success for all students. Students were intentionally matched in a peer-mentoring program, BUDDY-UP, on several factors, which included students' race and ethnicity (Black, Indigenous, and People of Color [BIPOC]) and gender, and whether they were first-generation. Mentors and mentees expressed satisfaction and derived benefits from the intentionally matched mentoring program. Participant comments demonstrated that these are not just extra supports but instead are fundamental to their essential everyday needs and ability to be successful both academically and socially. Incorporating support for all students needs to be a foundational aspect in the development of peer-mentoring programs to support the ability for all students to be successful in nursing education. [J Nurs Educ. 2022;61(11):633-635.].


Microtentacle Formation in Ovarian Carcinoma.
The development of chemoresistance to paclitaxel and carboplatin represents a major therapeutic challenge in ovarian cancer, a disease frequently characterized by malignant ascites and extrapelvic metastasis. Microtentacles (McTNs) are tubulin-based projections observed in detached breast cancer cells. In this study, we investigated whether ovarian cancers exhibit McTNs and characterized McTN biology. We used an established lipid-tethering mechanism to suspend and image individual cancer cells. We queried a panel of immortalized serous (OSC) and clear cell (OCCC) cell lines as well as freshly procured ascites and human ovarian surface epithelium (HOSE). We assessed by Western blot β-tubulin isotype, α-tubulin post-translational modifications and actin regulatory proteins in attached/detached states. We studied clustering in suspended conditions. Effects of treatment with microtubule depolymerizing and stabilizing drugs were described. Among cell lines, up to 30% of cells expressed McTNs. Four McTN morphologies (absent, symmetric-short, symmetric-long, tufted) were observed in immortalized cultures as well as ascites. McTN number/length varied with histology according to metastatic potential. Most OCCC overexpressed class III ß-tubulin. OCCC/OSC cell lines exhibited a trend towards more microtubule-stabilizing post-translational modifications of α-tubulin relative to HOSE. Microtubule depolymerizing drugs decreased the number/length of McTNs, confirming that McTNs are composed of tubulin. Cells that failed to form McTNs demonstrated differential expression of α-tubulin- and actin-regulating proteins relative to cells that form McTNs. Cluster formation is more susceptible to microtubule targeting agents in cells that form McTNs, suggesting a role for McTNs in aggregation. McTNs likely participate in key aspects of ovarian cancer metastasis. McTNs represent a new therapeutic target for this disease that could refine therapies, including intraperitoneal drug delivery.


Emerging tissue engineering strategies for annulus fibrosus therapy.
Low back pain is a major public health concern experienced by 80% of the world's population during their lifetime, which is closely associated with intervertebral disc (IVD) herniation. IVD herniation manifests as the nucleus pulposus (NP) protruding beyond the boundaries of the intervertebral disc due to disruption of the annulus fibrosus (AF). With a deepening understanding of the importance of the AF structure in the pathogenesis of intervertebral disc degeneration, numerous advanced therapeutic strategies for AF based on tissue engineering, cellular regeneration, and gene therapy have emerged. However, there is still no consensus concerning the optimal approach for AF regeneration. In this review, we summarized strategies in the field of AF repair and highlighted ideal cell types and pro-differentiation targeting approaches for AF repair, and discussed the prospects and difficulties of implant systems combining cells and biomaterials to guide future research directions. STATEMENT OF SIGNIFICANCE: Low back pain is a major public health concern experienced by 80% of the world's population during their lifetime, which is closely associated with intervertebral disc (IVD) herniation. However, there is still no consensus concerning the optimal approach for annulus fibrosus (AF) regeneration. In this review, we summarized strategies in the field of AF repair and highlighted ideal cell types and pro-differentiation targeting approaches for AF repair, and discussed the prospects and difficulties of implant systems combining cells and biomaterials to guide future research directions.


Epithelioid sarcoma presenting as the reflex sympathetic dystrophy syndrome.
A case of reflex sympathetic dystrophy caused by an epithelioid sarcoma is presented. This is the first report of a local peripheral tumour associated with the reflex sympathetic dystrophy syndrome.


Chronic kidney disease, insulin resistance, and incident diabetes in older adults.
Insulin resistance is a complication of advanced CKD. Insulin resistance is less well characterized in earlier stages of CKD. The response of the pancreatic β cell, effects on glucose tolerance, and risk of diabetes are not clear. The Cardiovascular Health Study included 4680 adults without baseline diabetes. The Chronic Kidney Disease Epidemiology Collaboration creatinine equation was used to obtain the estimated GFR (eGFR). Insulin resistance was evaluated as fasting insulin concentration. The insulin sensitivity index, β cell function, and glucose tolerance were assessed by oral glucose tolerance testing. Incident diabetes was defined as fasting glucose ≥126 mg/dl, nonfasting glucose ≥200 mg/dl, or use of glucose-lowering medications. Mean age was 72.5 years (range, 65-98 years). Mean eGFR was 72.2 (SD 17.1) ml/min per 1.73 m(2). After adjustment, each 10 ml/min per 1.73 m(2) lower eGFR was associated with a 2.2% higher fasting insulin concentration (95% confidence interval [CI], 1.4%, 2.9%; P<0.001) and a 1.1% lower insulin sensitivity index (95% CI, 0.03%, 2.2%; P=0.04). Surprisingly, eGFR was associated with an augmented β cell function index (P<0.001), lower 2-hour glucose concentration (P=0.002), and decreased risk of glucose intolerance (P=0.006). Over a median 12 years' follow-up, 437 participants (9.3%) developed diabetes. eGFR was not associated with the risk of incident diabetes. Among older adults, lower eGFR was associated with insulin resistance. However, with lower eGFR, β cell function was appropriately augmented and risks of impaired glucose tolerance and incident diabetes were not increased.


Dopamine output upon death reflects intraneuronal aspects while alive: accumulation of releasable dopamine during tetrodotoxin perfusion.
Intraneuronal and extracellular dopamine (DA) metabolism in rat striatum during inhibition of neurotransmission processes by tetrodotoxin (TTX) perfusion was examined by analysis of the striatal DA content and the amount of initial post-mortem DA output. In spite of there being an excess amount of DA already present in the striatum, the contents of DA and its metabolites were increased time-dependently by TTX perfusion through a microdialysis membrane. A massive increase in extracellular level of DA occurred immediately after death. The amount of the post-mortem DA output was further increased in proportion to the preceding TTX perfusion time. The ratio of the amount of post-mortem DA output to the striatal DA content was also increased by previous perfusion with TTX. Verapamil, a Ca2+ channel blocker, delayed the appearance of the peak of the DA output. These results suggest that releasable DA accumulates in neurons during TTX perfusion, and that the early part of the DA output upon death reflects intraneuronal aspects of DA metabolism while the animal is alive.


Transcriptional response of BmToll9-1 and RNAi machinery genes to exogenous dsRNA in the midgut of Bombyx mori.
Injection of dsRNA is widely applied to silence endogenous genes and study gene function in insects. However, it is not yet clear to what extent it can also exert non-specific effects, for instance by interference with the innate immune response. In this study, we report on the transcriptional response of BmToll9-1 to lipopolysaccharide (LPS) and dsRNA in the silkmoth, Bombyx mori. BmToll9-1 encodes a Toll receptor highly expressed in midgut tissue and that shows limited similarity to the mammalian TLR3 endolysosome receptor for dsRNA; while Dcr2 and Ago2 encode two key components of the RNAi machinery. An expression pattern study of all 14 Toll receptors in B. mori showed that BmToll9-1 was expressed in different larval and pupal tissues with the highest expression level detected in the midgut, indicating a possible function in immunity against pathogens taken up by the food. In order to investigate the response of BmToll9-1, different ways to deliver dsRNA, specific for GFP (dsGFP), and LPS were applied in Bombyx 5th instar larvae. The feeding experiments suggested that dsGFP did not suppress the expression of BmToll9-1 significantly, while LPS could suppress the expression of BmToll9-1 after 3h of feeding. On the other hand, the injection experiments showed that dsGFP, as well as LPS, could significantly inhibit the expression of BmToll9-1 in 3h. Bacteria that constantly expressed dsGFP could also down-regulate the expression of BmToll9-1 to a greater extent than bacteria that do not express dsGFP. The failure of dsGFP by feeding to affect the expression of BmToll9-1 was correlated with the rapid degradation of dsGFP by dsRNase in the midgut juice. Expression of the RNAi machinery genes Dcr2 and Ago2, as well as dsRNase, was also affected by injection of dsRNA and not by feeding, but in these cases an induction was observed instead of a down-regulation. Because LPS is a well-known pathogen-associated molecular pattern (PAMP), it suggested that the decrease in BmToll9-1 expression is a consequence of the activation of the innate immune response by LPS. The similar response of BmToll9-1 between the two triggers, LPS and dsRNA, suggests that dsRNA can also act as a PAMP in the midgut of Bombyx. Furthermore, induction of the genes Dcr2, Ago2 and dsRNase may also constitute a defense mechanism against invading dsRNA.


Androgen binding sites on nuclear matrix of normal and hyperplastic human prostate.
To further characterize human prostatic androgen receptor, nuclei were isolated from normal prostate (no. = 3) and benign prostatic hyperplasia specimens (no. = 10). High ionic strength (0.6 M KCl) treatment of nuclei released nuclear extractable androgen receptor and DNase I digestion then yielded nuclear matrices. Androgen receptor was quantified in the nuclear extract and nuclear matrix preparations by Scatchard analysis of specific R1881 binding. Only 1 of the 3 normal tissues had extractable androgen receptor (113 fmol. per gm. of tissue) while the mean concentration of extractable androgen receptor for BPH was 189 fmol. per gm. of tissue. The mean concentrations of matrix-bound androgen receptor were 325 fmol. per gm. of tissue and 548 fmol. per gm. of tissue for normal and hyperplastic prostate, respectively. The androgen binding sites on nuclear matrix may represent the functional intranuclear androgen receptor and a characterization of these sites may provide an understanding of the etiology of BPH.


Pathways for alpha-D-ribose utilization for nucleobase salvage and 5-fluorouracil activation in rat brain.
Recently, interest has increased in the use of alpha-D-ribose (Rib) as a naturally occurring nutriceutical for enhancement of cardiac and muscular performance. Most likely the elevation of available PRPP, following Rib administration, plays a key role in the salvage of purine nucleobases, thus, accelerating the restitution of ATP pool. In addition, administration of Rib improves some of the neurological symptoms in patients with adenylosuccinase deficiency. In this paper, we show that rat brain extract can catalyze the Rib-mediated salvage of both adenine and uracil, as well as the activation of the pyrimidine pro-drug, 5-fluorouracil (5-FU). The results strongly support that the pentose may be converted to both PRPP and Rib1-P for the salvage of the adenine and uracil, respectively. Most likely two-reaction pathway, composed of ribokinase and PRPP synthetase, is responsible of the PRPP formation, needed to salvage adenine to adenine nucleotides. A two-reaction pathway, composed of ribokinase and phosphopentomutase, appears to be responsible of the Rib1-P formation, needed to salvage uracil to uracil-nucleotides and to activate 5-FU to cytotoxic 5-FU-ribonucleotides. alpha-D-2'-Deoxyribose (deoxyRib) has a negligible effect on both the salvage of natural nucleobases to deoxyribonucleotides and on the activation of 5-FU to cytotoxic 5-FU-deoxynucleotides.


Enhancement of sonochemical production of hydroxyl radicals from pulsed cylindrically converging ultrasound waves.
Sonochemistry is the use of ultrasound to generate highly reactive radical species through the inertial collapse of a gas/vapour cavity and is a green alternative for hydrogen production, wastewater treatment, and chemical synthesis and modifications. Yet, current sonochemical reactors often are limited by their design, resulting in low efficacy and yields with slow reaction kinetics. Here, we constructed a novel sonochemical reactor design that creates cylindrically converging ultrasound waves to create an intense localised region of high acoustic pressure amplitudes (15 MPaPKPK) capable of spontaneously nucleating cavitation. Using a novel dosimetry technique, we determined the effect of acoustic parameters on the yield of hydroxyl radicals (HO), HO production rate, and ultimately the sonochemical efficiency (SE) of our reactor. Our reactor design had a significantly higher HO production rate and SE compared to other conventional reactors and across literature.


Integrating Activities of Laminins that Drive Basement Membrane Assembly and Function.
Studies on extracellular matrix proteins, cells, and genetically modified animals have converged to reveal mechanisms of basement membrane self-assembly as mediated by γ1 subunit-containing laminins, the focus of this chapter. The basic model is as follows: A member of the laminin family adheres to a competent cell surface and typically polymerizes followed by laminin binding to the extracellular adaptor proteins nidogen, perlecan, and agrin. Assembly is completed by the linking of nidogen and heparan sulfates to type IV collagen, allowing it to form a second stabilizing network polymer. The assembled matrix provides structural support, anchoring the extracellular matrix to the cytoskeleton, and acts as a signaling platform. Heterogeneity of function is created in part by the isoforms of laminin that vary in their ability to polymerize and to interact with integrins, dystroglycan, and other receptors. Mutations in laminin subunits, affecting expression or LN domain-specific functions, are a cause of human diseases that include those of muscle, nerve, brain, and kidney.


Effects of polygenic risk for suicide attempt and risky behavior on brain structure in young people with familial risk of bipolar disorder.
Bipolar disorder (BD) is associated with a 20-30-fold increased suicide risk compared to the general population. First-degree relatives of BD patients show inflated rates of psychopathology including suicidal behaviors. As reliable biomarkers of suicide attempts (SA) are lacking, we examined associations between suicide-related polygenic risk scores (PRSs)-a quantitative index of genomic risk-and variability in brain structures implicated in SA. Participants (n = 206; aged 12-30 years) were unrelated individuals of European ancestry and comprised three groups: 41 BD cases, 96 BD relatives ("high risk"), and 69 controls. Genotyping employed PsychArray, followed by imputation. Three PRSs were computed using genome-wide association data for SA in BD (SA-in-BD), SA in major depressive disorder (SA-in-MDD) (Mullins et al., 2019, The American Journal of Psychiatry, 176(8), 651-660), and risky behavior (Karlsson Linnér et al., 2019, Nature Genetics, 51(2), 245-257). Structural magnetic resonance imaging processing employed FreeSurfer v5.3.0. General linear models were constructed using 32 regions-of-interest identified from suicide neuroimaging literature, with false-discovery-rate correction. SA-in-MDD and SA-in-BD PRSs negatively predicted parahippocampal thickness, with the latter association modified by group membership. SA-in-BD and Risky Behavior PRSs inversely predicted rostral and caudal anterior cingulate structure, respectively, with the latter effect driven by the "high risk" group. SA-in-MDD and SA-in-BD PRSs positively predicted cuneus structure, irrespective of group. This study demonstrated associations between PRSs for suicide-related phenotypes and structural variability in brain regions implicated in SA. Future exploration of extended PRSs, in conjunction with a range of biological, phenotypic, environmental, and experiential data in high risk populations, may inform predictive models for suicidal behaviors.


IMP3 predicts aggressive superficial urothelial carcinoma of the bladder.
In this study, we investigated whether an oncofetal protein, IMP3, can serve as a new biomarker to predict progression and metastasis of early-stage urothelial carcinoma of the bladder. The expression of IMP3 in 242 patients with primary superficial bladder urothelial carcinoma and metastatic urothelial carcinoma was evaluated by immunohistochemistry. Patients with primary superficial urothelial carcinoma of the bladder were further investigated by use of survival analysis. Twenty percent (42 of 214) of primary superficial urothelial carcinomas and 93% (26 of 28) of metastatic urothelial carcinomas expressed IMP3. Kaplan-Meier plots and log-rank tests showed that patients with IMP3-positive tumors had a much lower progression-free survival (P = 0.0002) and disease-free survival rate (P = 0.0067) than did those with IMP3-negative tumors. The 5-year progression-free and disease-free survival rates were 91% and 94% in IMP3-negative patients versus 64% and 76% in IMP3-positive patients, respectively. Sixty percent of IMP3-positive patients with superficial invasive urothelial carcinoma at initial diagnosis went on to develop metastases, whereas no metastasis was found in IMP3-negative patients (P = 0.0017). In the multivariable Cox analysis, patients with IMP3 expression in their superficial urothelial carcinomas subsequently developed invasive tumors or metastasis at a rate that was about five times greater than cases without expression of IMP3 adjusting for other well-known clinical variables (tumor stage and grade, etc.). Our findings indicate that IMP3 is an independent prognostic marker that can identify a group of patients with a high potential to develop progression and who might benefit from early aggressive therapy.