Myricetin Exerts Anti-Obesity Effects through Upregulation of SIRT3 in Adipose Tissue.
Myricetin is a biologically active natural polyphenol with beneficial effects on metabolic health. This study aimed to examine the effects of myricetin on the expression levels of genes involved in lipolysis and mitochondrial respiration in adipocytes and the anti-obesity potential of myricetin. The results indicated that myricetin reduced triglyceride (TG) content and increased mitochondrial content and oxygen consumption rate (OCR) in adipocytes in vitro. To determine anti-obesity effect of myricetin, C57BL6/J mice were fed a high-fat diet (HFD) for eight weeks and then treated with myricetin (10 mg/kg) for 2 weeks. The in vivo treatment of myricetin reduced body weight by 11%. Furthermore, it improved the glucose tolerance, and increased fatty acid consumption of HFD-fed mice. Myricetin treatment increased Sirt3 expression and reduced the acetylation of mitochondrial proteins in adipose tissue. Finally, the knockdown of Sirt3 in adipocytes reduced the myricetin-induced increase in mitochondrial oxygen consumption rate by about 27% compared to controls. Our results indicated that myricetin exerted anti-obesity effects through the upregulation of Sirt3 expression and mitochondrial metabolism in adipose tissue.


Individual prostheses and resection templates for mandibular resection and reconstruction.
This new technique uses helical computed tomography data and computer-aided design and manufacturing for preoperative fabrication of individual mandibular prostheses together with corresponding resection templates. Coherent 3D geometries for computer-based models are the basis for the construction of prostheses and provide data for a computerized numerical control fabrication. Fixation plates are fabricated with the titanium prostheses. The identical data of these plates are used for the computer-aided design and manufacturing of resection templates, which guide an oscillating saw in a precisely determined resection plane. This plane again is identical with the prostheses' margins for mandibular body replacement. The use of this technique in four patients is reported on: after temporary insertion of the templates for resection and after resection, the prostheses were stabilized with the same screws in the same screw-holes where the templates had been. Resection and reconstruction were thus highly precise, safe and fast and primarily led to excellent aesthetic and functional results. Wound-healing depends on a safe soft-tissue reconstruction over these large prostheses. Coverage with flaps seems obligatory. In spite of the superior technical aspects, the clinical long-term results of this new technique were poor.


Complementary and alternative medicine use among Asian Indians in the United States: a national study.
Asian Americans use complementary and alternative medicine (CAM) at a higher rate than Americans. This national study on Asian Indians (AIs), the third largest Asian-American subgroup in the United States, examined the CAM use by gender and its association with acculturation, health behaviors, and access to health care. This was a cross-sectional survey. The subjects consisted of 1824 AI adults in six states with higher concentration of AIs. Mean age and years lived in the United States was 45.7 +/- 12.8 and 16.6 +/- 11.1 years, respectively. The respondent majority was male, immigrants, college graduates, and had access to care. Sixty three percent (63%) of AIs used at least one type of CAM; most common was a vegetarian diet, followed by use of dietary and herbal supplements and alternative medical systems. Females reported a significantly higher use of CAM, a vegetarian diet, and use of dietary and herbal supplements than AI males. Older age, female gender, having no access to care, and spirituality predicted CAM use in the logistic regression model. Older age, female gender, unmarried, and higher income was associated with use of dietary and herbal supplements; AIs who reported being vegetarian were more likely to be female, unmarried, spiritual, and self-reported their physical health to be fair or poor. This is the first national study of CAM use among AIs by gender and selected respondent characteristics. Results provide important information on health behaviors, beliefs, and patterns of CAM use in this ethnic subgroup to be factored into patient education.


[The adaptogenic effects of hyperbaric oxygenation within the complex therapy of inflammation diseases of the internal female genitals].
The effects of hyperbaric oxygenation (HBO), when used jointly with the routine complex therapy, made in the clinical signs of inflammatory diseases of the internal female genitals as well as in the lipid peroxidation/antioxidant system were investigated within the case study. The reaction of the antioxidant system to the HBO action was found to depend on a degree of its original exhaustion. An opinion was put forward on the need in the individual approach to whether to add the HBO method to the complex therapy or not. If the reaction to the hyperbaric effects is positive, the HBO method must be used to the full extent. If there is progressing of the exhaustion degree of the antioxidant system, the HBO course must be interrupted.


Do Baseline Executive Functions Mediate Prospective Memory Performance under a Moderate Dose of Alcohol?
Prospective memory (PM) is memory for delayed intentions. While deleterious effects of acute doses of alcohol on PM have been documented previously using between-subjects comparisons, the current study adopted a single blind placebo-controlled within-subjects design to explore whether the extent to which alcohol-related impairments in PM are mediated by executive functions (EFs). To this end, 52 male social drinkers with no history of substance-related treatment were tested using two parallel versions of a clinical measure of PM (the Memory for Intentions Test; Raskin et al., 2010), and a battery of EF measures. Testing took place on two occasions, with the order of administration of the alcohol and placebo conditions being fully counterbalanced. Overall, PM was worse under alcohol and participants showed deficits on five of the six subscales making up the clinical test. Hierarchical multiple regression analyses demonstrated that EFs did not predict PM performance decrements overall but did predict performance when time cues were presented and when verbal responses were required. Phonemic fluency was the strongest of the EF predictors; a greater capacity to gain controlled access to information in long-term memory predicted a smaller difference between placebo- and alcohol-related performance on both the time cue and verbal response scales. PM is crucial to compliance with, and response to, both therapy programs and alcohol harm prevention campaigns. The results indicate that individual differences in cognitive function need to be taken into account when designing such interventions in order to increase their effectiveness.


Berbamine induced activation of the SIRT1/LKB1/AMPK signaling axis attenuates the development of hepatic steatosis in high-fat diet-induced NAFLD rats.
Non-alcoholic fatty liver disease (NAFLD), a chronic metabolic disorder is concomitant with oxidative stress and inflammation. This study aimed to assess the effects of berbamine (BBM), a natural bisbenzylisoquinoline alkaloid with manifold biological activities and pharmacological effects on lipid, cholesterol and glucose metabolism in a rat model of NAFLD, and to explicate the potential mechanisms underlying its activity. BBM administration alleviated the increase in the body weight and liver index of HFD rats. The aberrations in liver function, serum parameters, and microscopic changes in the liver structure of HFD fed rats were significantly improved upon BBM administration. BBM also significantly attenuated oxidative damage and inhibited triglyceride and cholesterol synthesis. The SIRT1 deacetylase activity was also enhanced by BBM through liver kinase B1 and activated AMP-activated protein kinase. Activation of the SIRT1/LKB1/AMPK pathway prevented the downstream target ACC (acetyl-CoA carboxylase) and elevation in the expression of FAS (fatty acid synthase) and SCD1 (steroyl CoA desaturase). BBM also modulated the expression of PPARs maintaining the fatty acid homeostasis regulation. The assessment of berbamine induced ultrastructural changes by TEM analysis and the expression of autophagic markers LC3a/b, Beclin 1 and p62 revealed the induction of autophagy to alleviate fatty liver conditions. These results show novel findings that BBM induced protection against hepatic lipid metabolic disorders is achieved by regulating the SIRT1/LKB1/AMPK pathway, and thus it emerges as an effective phyoconstituent for the management of NAFLD.


Barriers to Seeking and Accepting Treatment for Perinatal Depression: A Qualitative Study in Rio de Janeiro, Brazil.
Few studies have investigated the reasons why pregnant and puerperal women fail to seek or accept treatment for perinatal depression in low- and middle-income countries, where there is a high prevalence of this disorder. To help fill this gap, this study investigated the factors influencing the decision not to seek or to refuse treatment for perinatal depression in a low-income community in Rio de Janeiro, Brazil. Qualitative research was conducted in two primary health care units in Rio de Janeiro, Brazil in 2017-2018. Five focus groups were held with 26 women. Convenience sampling was used, and the sample size was determined by data saturation. A content analysis methodology was used to identify theme categories to objectively describe the group's manifest contents. Ten categories were obtained: stigma and misconception, self-image as a mother, socioeconomic stigma, lack of knowledge, lack of a health service approach to mental health, difficulty recognising depression symptoms, fear of children being removed, negative reaction to patient referral, denial of the problem and previous experience with the care unit. Perinatal depression is permeated with stigma and prejudice, and there is a belief that women with depression are unable to be good mothers. It is important to conduct programmes disseminating information about perinatal depression and implementing an approach that includes routine consultations so that women can access perinatal mental health services.


Mammogenesis and induced lactation with or without reserpine in nulliparous dairy goats.
Nulliparous goats were used to evaluate the effects of a standard protocol for inducing lactation with or without using a prolactin-releasing agent (reserpine). Estrus was synchronized and goats were submitted to daily s.c. injections of estradiol-17beta and progesterone (0.5 and 1.25 mg/kg of body weight, respectively) for 7 d. The goats were divided into 2 groups and injected i.m. with 1 mg/d of reserpine (n = 7) or the vehicle (n = 7) on d 12, 14, 16, 18, and 20. Lactation was initiated by i.m. injections of dexamethasone (10 mg/d) from d 18 to 20. Goats were machine milked once daily from d 21 to 120, at which time they were mated with herd sires. Milk was measured and sampled daily during wk 1 of lactation and weekly thereafter. Udder traits were measured in all goats at d -2 (before the induction treatment) and on d 35 and 100 (during lactation). Goats initiated lactation on d 21 (100%) and milk yield increased thereafter. The milk yield of control and reserpine-treated goats increased as lactation advanced, peaking at wk 10 of lactation, when reserpine-treated goats yielded 1,079 +/- 89 mL/d of milk compared with 850 +/- 96 mL/d for control goats. Yet milk yield at the peak was only 55% of the peak milk yield observed in contemporary primiparous goats. The composition of initial milk (d 21) was different from that expected for colostrum. Milk composition stabilized after d 3 of lactation. There were no differences among groups for milk fat, protein, casein, or whey protein, but milk from control goats contained greater nonprotein nitrogen than that from reserpine-treated goats (0.48 +/- 0.02 vs. 0.41 +/- 0.02%). Teat length increased from 24.7 +/- 1.1 to 34.5 +/- 2.4 mm in control goats during mammogenesis (d -2 to 35), but stabilized in reserpine goats (25.2 +/- 2.2 mm). The distance between teats (11.5 +/- 0.4 cm), and the volume (922 +/- 63 mL) and depth (15.6 +/- 0.60 cm) of the udder increased similarly in both groups during mammogenesis and lactation. After mating, 82% of herdmates became pregnant, whereas only 21% of the lactation-induced goats conceived (1 reserpine-treated and 2 control goats). In conclusion, lactation induction was effective in nulliparous goats, but neither milk yield nor the side effects on fertility seem to support its recommendation.


Multidrug-resistant tumor cells remain sensitive to a recombinant interleukin-4-Pseudomonas exotoxin, except when overexpressing the multidrug resistance protein MRP1.
Tumor cells may become resistant to conventional anticancer drugs through the occurrence of transmembrane transporter proteins such as P-glycoprotein (ABCB1), breast cancer resistance protein (ABCG2), or members of the multidrug resistance-associated protein family (MRP1-MRP5; ABCC1-ABCC5). In this report, we studied whether tumor cells that are cytostatic drug resistant because of overexpression of one of the above mentioned proteins are sensitive to a new anticancer agent, interleukin-4 toxin (IL-4 toxin). IL-4 toxin is a fusion protein composed of circularly permuted IL-4 and a truncated form of Pseudomonas exotoxin (PE) [IL-4(38-37)-PE38KDEL]. Ninety-six-h cytotoxicity assays and 10-day clonogenic assays showed that drug-selected multidrug resistant (MDR) tumor cells that overexpress P-glycoprotein or breast cancer resistance proteins are still sensitive to IL-4 toxin. Also, tumor cells transfected with cDNA for MRP2-5 showed no resistance, or marginal resistance, only to the toxin as compared with the parent cells. In contrast, MRP1-overexpressing cells, both drug selected and MRP1 transfected, are clearly resistant to IL-4 toxin with resistance factors of 4.3 to 8.4. MRP1-overexpressing cells were not resistant to PE itself. IL-4 toxin resistance in MRP1-overexpressing cells could be reversed by the MRP1 inhibitors probenecid or MK571 and were not affected by glutathione depletion by DL-buthionine-S,R-sulfoximine. In a transport assay using plasma membrane vesicles prepared from MRP1-overexpressing cells, IL-4 toxin and IL-4, but not PE, inhibited the translocation of the known MRP1 substrate 17beta-estradiol 17-(beta-D-glucuronide) (E(2)17betaG). These data suggest that MRP1-overexpressing cells are resistant to IL-4 toxin because of extrusion of this agent by MRP1. Still, the results of this study demonstrate that IL-4 toxin effectively kills most MDR tumor cells and, therefore, represents a promising anticancer drug.


C-peptide increases the expression of vasopressin-activated calcium-mobilizing receptor gene through a G protein-dependent pathway.
Although an increasing number of reports suggest that physiological concentrations of C-peptide protect against the development of diabetic nephropathy, possibly through the modulation of Na-K pump activity, the intracellular pathways controlled by C-peptide are still unrecognized. C-peptide and vasopressin share similar intracellular effects including the activation of calcium influx and endothelial nitric oxide synthase. Both hormones stimulate also the activity of Na-K pump activity. Whether the activity of C-peptide is mediated by the recently identified vasopressin-activated calcium-mobilizing receptor (VACM-1) has never been previously investigated. To clarify this issue, we evaluated the effect of C-peptide on VACM-1 RNA (measured by semiquantitative RT-PCR) and protein expression (measured by immunoblotting) in human skin fibroblasts (where a specific binding of C-peptide was demonstrated) and in human mesangial cells, the cellular target of diabetic nephropathy. C-peptide-induced activation of VACM-1 was demonstrated in fibroblasts from six healthy individuals (0.51+/-0.1 vs 1.48+/-0.4, arbitrary units+/-s.e., P = 0.025). This finding was paralleled by an increased VACM-1 protein expression (5.64+/-1.0 vs 8.47+/-1.2, arbitrary units+/-s.e., P= 0.043). Similar results were confirmed in three independent cultures of human mesangial cells. VACM-1 activation in fibroblasts was insensitive to phosphatidylinositol-3-kinase inhibitor LY294002, but was inhibited by pertussis toxin, suggesting that activation of VACM-1 could be mediated by a G protein-coupled receptor. This study demonstrates for the first time that C-peptide activates VACM-1, possibly through a G protein-coupled receptor. Further studies are needed to clarify whether VACM-1 is involved in the protective effect of C-peptide against the development of diabetic nephropathy.


Significant Impact of Coronavirus Disease 2019 (COVID-19) on Human Immunodeficiency Virus (HIV) Care in Hospitals Affecting the First Pillar of the HIV Care Continuum.
During COVID-19 lockdown, the in-hospital number of HIV indicator conditions decreased disproportionally compared with other non-COVID-19 diseases, which was accompanied by reduced HIV testing rates, number and proportion of positive HIV tests, and new HIV referrals, with more late presentation after lockdown cessation, indicating a significantly impacted HIV care continuum.


Identification of novel methylation markers in hepatocellular carcinoma using a methylation array.
Promoter CpG island hypermethylation has become recognized as an important mechanism for inactivating tumor suppressor genes or tumor-related genes in human cancers of various tissues. Gene inactivation in association with promoter CpG island hypermethylation has been reported to be four times more frequent than genetic changes in human colorectal cancers. Hepatocellular carcinoma is also one of the human cancer types in which aberrant promoter CpG island hypermethylation is frequently found. However, the number of genes identified to date as hypermethylated for hepatocellular carcinoma (HCC) is fewer than that for colorectal cancer or gastric cancer, which can be attributed to fewer attempts to perform genome-wide methylation profiling for HCC. In the present study, we used bead-array technology and coupled methylation-specific PCR to identify new genes showing cancer-specific methylation in HCC. Twenty-four new genes have been identified as hypermethylated at their promoter CpG island loci in a cancer-specific manner. Of these, TNFRSF10C, HOXA9, NPY, and IRF5 were frequently hypermethylated in hepatocellular carcinoma tissue samples and their methylation was found to be closely associated with inactivation of gene expression. Further study will be required to elucidate the clinicopathological implications of these newly found DNA methylation markers in hepatocellular carcinoma.


Selenium effects on arsenic cytotoxicity and protein phosphorylation in human kidney cells using chip-based nanoLC-MS/MS.
Although arsenic toxicity is well known, little is known of how it exerts its effects at the proteome level. Protein phosphorylation is an important post-translational modification in the regulation of cell signaling. Despite the importance of protein phosphorylation, the identification and characterization of phosphorylated proteins, as influenced by interaction between arsenic and selenium species have not been fully studied. The aim of this study is to identify phosphorylation in arsenic toxified cells, with and without selenium present. Here, we identify the phosphorylated proteins related to post translational modifications (PTMs) after inorganic arsenic (iAs) and selenomethionine (SeMet) were inoculated together with HEK293 human kidney cells. In this study, using TiO(2)-based nanoLC-phosphochip® coupled to ESI-MS we observed phosphorylated peptide enrichment and significant reduction in sample complexity. The identification of phosphorylated proteins in highly complex digests of cell lysate were markedly different with As toxification only, or when in the presence of SeMet. Several phosphorylation sites and proteins are identified using Spectrum Mill and Mascot protein data base search engines. Cytotoxicity studies showed that SeMet significantly reduces the cytotoxic effect of iAs in HEK293 cells, while inorganic selenium did not.


Topoisomerase IIbeta is required for lamina-specific targeting of retinal ganglion cell axons and dendrites.
The specific partnering of synaptically connected neurons is central to nervous system function. Proper wiring requires the interchange of signals between a postmitotic neuron and its environment, a distinct pattern of transcription in the nucleus, and deployment of guidance and adhesion cues to the cell surface. To identify genes involved in neurite targeting by retinal ganglion cells (GCs), their presynaptic partners in the retina, and their postsynaptic targets in the optic tectum, we undertook a forward genetic screen for mutations disrupting visual responses in zebrafish. This rapid primary screen was subsequently refined by immunohistochemical labeling of retinal and tectal neurites to detect patterning errors. From this unbiased screen, the notorious (noto) mutant exhibited the most specific phenotypes: intact retinal and tectal differentiation but multiple neurite targeting defects in the retinal inner plexiform layer (IPL) and tectal neuropil. Positional cloning and morpholino phenocopy revealed that the mutation disrupts Topoisomerase IIβ (Top2b), a broadly distributed nuclear protein involved in chromatin modifications during postmitotic differentiation. Top2b-DNA interactions are known to regulate transcription of developmentally important genes, including axon guidance factors and cell adhesion molecules, but a specific role in local synaptic targeting has not been previously described. The neurite targeting defects among GC axons are largely restricted to crossovers between sublaminae of a specific layer, SFGS, and were shown by mosaic analysis to be autonomous to the GC axons. The noto mutant provides the first example of the importance of an epigenetic regulator, Top2b, in the intricate series of events that lead to a properly wired visual system.


Availability of ambulance patient care reports in the emergency department.
Clinical handovers of patient care among healthcare professionals is vulnerable to the loss of important clinical information. A verbal report is typically provided by paramedics and documented by emergency department (ED) triage nurses. Paramedics subsequently complete a patient care report which is submitted electronically. This emergency medical system (EMS) patient care report often contains details of paramedic assessment and management that is not all captured in the nursing triage note. EMS patient care reports are often unavailable for review by emergency physicians and nurses. Two processes occur in the distribution of EMS patient care reports. The first is an external process to the ED that is influenced by the prehospital emergency medical system and results in the report being faxed to the ED. The second process is internal to the ED that requires clerical staff to distribute the fax report to accompany patient charts. A baseline audit measured the percentage of EMS patient care reports that were available to emergency physicians at the time of initial patient assessments and showed a wide variation in the availability of EMS reports. Also measured were the time intervals from patient transfer from EMS to ED stretcher until the EMS report was received by fax (external process measure) and the time from receiving the EMS fax report until distribution to patient chart (internal process measure). These baseline measures showed a wide variation in the time it takes to receive the EMS reports by fax and to distribute reports. Improvement strategies consisted of: 1. Educating ED clerical staff about the importance of EMS reports 2. Implementing a new process to minimize ED clerical staff handling of EMS reports for nonactive ED patients 3. Elimination of the automatic retrieval of old hospital charts and their distribution for ED patients 4. Introduction of an electronic dashboard for patients arriving by ambulance to facilitate more efficient distribution of EMS reports. Implementation of change strategies did not result in a significant improvement in the percentage of EMS reports available to emergency physicians at the time of initial patient assessment. However, tracking both external and internal processes that influence EMS report availability showed the internal process time from fax report receipt to distribution significantly improved. This improvement reflected the change strategies that were all directed at improving the internal process. EMS patient care reports are more efficiently processed and distributed in the ED due to change strategies implemented that targeted the ED's internal process of EMS report distribution. The external process responsible for transmitting EMS reports to the ED is the limiting factor that prevents consistent timely access of EMS reports by emergency physicians and will require dedicated improvement strategies.


The Response of Picea abies Somatic Embryos to UV-B Radiation Depends on the Phase of Maturation.
Ultraviolet-B (UV-B) radiation is a key environmental signal which initiates diverse responses that affect the metabolism, development, and viability of plants. In keeping with our previous studies, we concentrated primarily on how UV-B radiation affects Norway spruce [Picea abies (L.) Karst.] somatic embryo maturation and how phenolics and polyamines (PAs) are linked to the defense response invoked by UV-B irradiation. We treated clusters of Norway spruce embryogenic culture (EC) with UV-B during the five stages of embryo maturation (early, cylindrical, precotyledonary, cotyledonary, and mature embryos). For the first time, we take an advantage of the unique environmental scanning electron microscope AQUASEM II to characterize somatic embryos in their native state. The severity of the irradiation effect on embryonal cell viability was shown to be dependent on the intensity of radiation as well as the stage of embryo development, and might be related to the formation of protoderm. The response of early embryos was characterized by an increase in malondialdehyde (MDA), a marked decrease in PA contents and a decline in phenolics. The reduced ability to activate the defense system seems to be responsible not only for the severe cell damage and decrease in viability but also for the inhibition of embryo development. The significant reduction in spermidine (Spd), which has been reported to be crucial for the somatic embryo development of several coniferous species, may be causally linked to the limited development of embryos. The pronounced decrease in cell wall-bound ferulic acid might correspond to failure of somatic embryos to reach more advanced stages of development. Embryos at later stages of development showed stress defense responses that were more efficient against UV-B exposure.


DLC-1 down-regulation via exosomal miR-106b-3p exchange promotes CRC metastasis by the epithelial-to-mesenchymal transition.
Exosomes (Exo) have emerged as potent amplifiers of pro-tumorigenic signals to distant cells. The knowledge of their role in colorectal cancer (CRC) is continuously up-growing, although their contribution to metastasis remains largely unclear. Liu et al. (Clinical Science (2020) 134, https://doi.org/10.1042/CS20191087) in their work have described a novel mechanism by which CRC-derived Exo promote metastasis through the down-regulation of the deleted in liver cancer-1 (DLC-1), a gene involved in the epithelial-to-mesenchymal transition (EMT) event in cancer cells. The Authors also demonstrated an increase in serum exosomal miR-106b-3p in patients with metastatic CRC, suggesting its potential implication as a prognostic biomarker. These findings may be of great effort in clarifying the underlying mechanisms of CRC metastasis and provide new targets for future researches.


Effects of prochloraz on DNA damage, lipid peroxidation and antioxidant system in vitro.
Prochloraz is a broad-spectrum contact imidazol fungicide used against several diseases in wheat, barley and oleaginous plants but also for treatment of flower production. Although prochloraz has endocrine disrupting and hepatocarcinogenic effects, there is lack of data on toxic effects of prochloraz. Therefore, we aimed to investigate the DNA damage effects of prochloraz in NRK-52E cells by using Ames and Comet assay. By using a standard alkaline Comet assay procedure, there was no DNA damage observed after 24 h prochloraz exposure. It also showed that prochloraz caused neither base-pair substitution nor frame shift mutations by using TA98, TA100 strains, respectively, with/without metabolic activation in Ames assay. Both Comet and Ames assays, the exposure concentrations were 12.5, 25, 50 and 100 µM. IC50 value of prochloraz was determined as 110.76 µM in NRK-52E cells by MTT cytotoxicity test. Also, we evaluated possible effects of prochloraz on lipid peroxidation, reduced glutathione (GSH), oxidized glutathione (GSSG) and antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and glutathione reductase (GSH-Rd) in NRK-52E cells at 1-50 µM concentrations. Prochloraz induced lipid peroxidation and altered glutathione contents and antioxidant enzyme activities in NRK-52E cells. Our results indicated that prochloraz showed no evidence of mutagenicity and DNA damage; however, some alterations were observed on lipid peroxidation and antioxidant systems in prochloraz treatment.


Analysis of the spelling patterns of 4th grade students based on a word dictation task.
The purpose of this study was to establish a profile of the spelling patterns studied in students from public and private schools and to describe a word spelling task for clinical and educational settings. Eighty-two fourth grade students belonging to the elementary school of public and private schools in São Paulo, ranging in age from nine to ten years, took part in this study. The spelling task consisted of a list of ten high frequency words (HFW), ten low frequency words (LFW), and ten pseudowords (PW), in which the typology and number of spelling errors were described. To compare the average number of mistakes on the HFWs, LFWs, and PWs, we used an analysis of variance and Tukey's multiple comparisons (p<0.05). Using a cluster analysis, homogeneous groups were formed based on their performance. Results indicated that the average number of mistakes in the LFWs was higher than in the HFWs (p=0.000) and PWs (p=0.000), and the number of mistakes in the HFWs was lower than in the PWs (p=0.009). The highest number of mistakes was found in the following categories: "others", "rule generalization", "omission", "voiced-voiceless", and "addition". There were no mistakes of the type "ão-am" and "blend-separation" in the HFWs. Spelling errors are a part of the process of learning to write, and students can show some variance in spelling performance. Furthermore, students need to be stimulated to analyze words and their aspects of phonology, morphology, and semantics. An analysis from the types of errors is not enough to plan intervention programs, but instead is necessary to understand the strategies that the child uses to write.


Levofloxacin-imipenem combination prevents the emergence of resistance among clinical isolates of Pseudomonas aeruginosa.
A 2-compartment in vitro pharmacokinetic model (IVPM) was used to assess the potential of a levofloxacin-imipenem combination to prevent the emergence of resistance during treatment of Pseudomonas aeruginosa infection. Log-phase cultures (10(8) cfu/mL) of 3 clinical isolates were inoculated into the peripheral compartment of the IVPMs and were treated with simulated human doses of levofloxacin (750 mg) and imipenem (250 mg). Pharmacodynamics and the emergence of resistance were evaluated over the course of 24 h. Resistant mutants were evaluated for transcriptional expression of specific efflux pumps. Initially, rapid killing was observed in association with each regimen. However, with levofloxacin and imipenem alone, rapid regrowth was observed as a result of the selection of resistant subpopulations. Analysis of mutants selected by levofloxacin demonstrated that mexEF-oprN-overexpressing subpopulations resistant to both levofloxacin and imipenem were selected from cultures of all 3 strains. Nevertheless, the levofloxacin-imipenem combination rapidly eradicated all 3 P. aeruginosa strains. These data suggest that levofloxacin-imipenem may be an effective combination for preventing the emergence of resistance among P. aeruginosa strains, even when subpopulations resistant to both drugs are present. Further studies are warranted to evaluate the use of this combination against strains with established resistance to either or both drugs.


Inbreeding uncovers fundamental differences in the genetic load affecting male and female fertility in a butterfly.
Inbreeding depression is most pronounced for traits closely associated with fitness. The traditional explanation is that natural selection eliminates deleterious mutations with additive or dominant effects more effectively than recessive mutations, leading to directional dominance for traits subject to strong directional selection. Here we report the unexpected finding that, in the butterfly Bicyclus anynana, male sterility contributes disproportionately to inbreeding depression for fitness (complete sterility in about half the sons from brother-sister matings), while female fertility is insensitive to inbreeding. The contrast between the sexes for functionally equivalent traits is inconsistent with standard selection arguments, and suggests that trait-specific developmental properties and cryptic selection play crucial roles in shaping genetic architecture. There is evidence that spermatogenesis is less developmentally stable than oogenesis, though the unusually high male fertility load in B. anynana additionally suggests the operation of complex selection maintaining male sterility recessives. Analysis of the precise causes of inbreeding depression will be needed to generate a model that reliably explains variation in directional dominance and reconciles the gap between observed and expected genetic loads carried by populations. This challenging evolutionary puzzle should stimulate work on the occurrence and causes of sex differences in fertility load.


A review of gamma oscillations in healthy subjects and in cognitive impairment.
This review describes a wide range of functional correlates of gamma oscillations in whole-brain work, in neuroethology, sensory-cognitive dynamics, emotion, and cognitive impairment. This survey opens a new window towards understanding the brain's gamma activity. Gamma responses are selectively distributed in the whole brain, and do not reflect only a unique, specific function of the nervous system. Sensory responses from cortex, thalamus, hippocampus, and reticular formations in animal and human brains, and also cognitive responses, were described by several authors. According to reviewed results, it becomes obvious that cognitive disorders, and medication-which influence the transmitter release-change entirely the understanding of the big picture in cognitive processes. Gamma activity is evoked or induced by different sensory stimuli or cognitive tasks. Thus, it is argued that gamma-band synchronization is an elementary and fundamental process in whole-brain operation. In conclusion, reasoning and suggestions for understanding gamma activity are highlighted.


Sec1 regulates intestinal mucosal immunity in a mouse model of inflammatory bowel disease.
Inflammatory bowel disease (IBD) is a common immune-mediated condition with its molecular pathogenesis remaining to be fully elucidated. This study aimed to deepen our understanding of the role of FUT2 in human IBD, by studying a new surrogate gene Sec1, a neighboring gene of Fut2 and Fut1 that co-encodes the α 1,2 fucosyltransferase in mice. CRISPR/Cas9 was used to prepare Sec1 knockout (Sec1-/-) mice. IBD was induced in mice using 3% w/v dextran sulphate sodium. Small interfering RNA (siRNA) was employed to silence Sec1 in murine colon cancer cell lines CT26.WT and CMT93. IBD-related symptoms, colonic immune responses, proliferation and apoptosis of colon epithelial cells were assessed respectively to determine the role of Sec1 in mouse IBD. Impact of Sec1 on the expression of death receptor 5 (DR5) and other apoptosis-associated proteins were determined. Sec1 knockout was found to be associated with deterioration of IBD in mice and elevated immune responses in the colonic mucosa. Silencing Sec1 in CT26.WT and CMT93 cells led to greater secretion of inflammatory cytokines IL-1β, IL-6 and TNF-α. Cell counting kit 8 (CCK8) assay, flow cytometry and TUNEL detection suggested that Sec1 expression promoted the proliferation of colon epithelial cells, inhibited cell apoptosis, reduced cell arrest in G0/G1 phase and facilitated repair of inflammatory injury. Over-expression of DR5 and several apoptosis-related effector proteins was noticed in Sec1-/- mice and Sec1-silenced CT26.WT and CMT93 cells, supporting a suppressive role of Sec1 in cell apoptosis. Our results depicted important regulatory roles of Sec1 in mouse IBD, further reflecting the importance of FUT2 in the pathogenesis of human IBD.


Circadian Change in Blink Reflex Recovery in Restless Legs Syndrome.
Restless legs syndrome (RLS) is associated with dysfunction of the dopaminergic systems in the pathways that specifically link the sensory input and motor output. Keeping in mind that clinical symptomatology in RLS and cerebrospinal fluid dopamine concentrations in healthy individuals show changes throughout the day, we hypothesized that excitability of the related pathways increases during the night in RLS, and in the present study, we aimed to analyze our hypothesis by the assessment of blink reflex (BR) recovery. Eleven patients with primary RLS and eight age- and gender-matched healthy subjects were included in the study. All participants underwent detailed interviews and neurological examinations. BR responses were recorded after single and paired supraorbital stimulation during the early afternoon and late at night. For double stimulation, interstimulus intervals (ISI) of 100, 300, and 500 ms were used. Daytime and nighttime investigations were separately compared between the patient and control groups (between-group analyses). In-group analyses were conducted between daytime and nighttime investigations of the patient and control groups. BR responses to single stimuli were normal in all participants at all sessions. R2 recovery was the highest in the patient group during nighttime investigations. In-group analysis showed a reduction of R2 recovery during the night in healthy subjects. R2 recoveries at ISIs of 300 and 500 ms at nighttime were higher in RLS patients but did not reach statistical significance. The BR circuit is less excitable during the night in healthy individuals, whereas the reduction of excitability is lost in RLS. Despite the limited number of included subjects, we suggest that the normal circadian modulation of the BR circuit is lost in RLS.


From mechanical folding trajectories to intrinsic energy landscapes of biopolymers.
In single-molecule laser optical tweezer (LOT) pulling experiments, a protein or RNA is juxtaposed between DNA handles that are attached to beads in optical traps. The LOT generates folding trajectories under force in terms of time-dependent changes in the distance between the beads. How to construct the full intrinsic folding landscape (without the handles and beads) from the measured time series is a major unsolved problem. By using rigorous theoretical methods--which account for fluctuations of the DNA handles, rotation of the optical beads, variations in applied tension due to finite trap stiffness, as well as environmental noise and limited bandwidth of the apparatus--we provide a tractable method to derive intrinsic free-energy profiles. We validate the method by showing that the exactly calculable intrinsic free-energy profile for a generalized Rouse model, which mimics the two-state behavior in nucleic acid hairpins, can be accurately extracted from simulated time series in a LOT setup regardless of the stiffness of the handles. We next apply the approach to trajectories from coarse-grained LOT molecular simulations of a coiled-coil protein based on the GCN4 leucine zipper and obtain a free-energy landscape that is in quantitative agreement with simulations performed without the beads and handles. Finally, we extract the intrinsic free-energy landscape from experimental LOT measurements for the leucine zipper.


Hepatoprotective standardized EtOH-water extract of the leaves of Ziziphus jujuba.
The hepatoprotective activity of the EtOH-water extract of Ziziphus jujuba leaves was evaluated against CCl4-induced hepatic damage in mice. The EtOH-water extract significantly alleviated liver damage as indicated by the decreased levels of serum ALT and AST and the decreased MDA content, the increased levels of SOD, GSH and GSH-Px, and the reduced pathological tissue injury induced by CCl4. A quantitative analysis of fifteen major constituents (1-15) of the EtOH-water extract of the leaves of Z. jujuba was conducted by HPLC-DAD. Based on our research results, it can be concluded that the EtOH-water extract of the leaves of Z. jujuba is efficacious for prevention and treatment of CCl4-induced hepatic injury in mice. Flavonoids might be the active ingredients responsible for the biological and pharmacological activities towards hepatoprotection.


Disseminated histoplasmosis in an Atlantic bottlenose dolphin (Tursiops truncatus).
An approximately 37-yr-old female Atlantic bottlenose dolphin (Tursiops truncatus) died after a 4-mo illness characterized by intermittent anorexia, lethargy, mild neutrophilic leukocytosis, and mild nonregenerative anemia. At necropsy, the lungs were diffusely consolidated, and histopathology of the lungs revealed severe pneumonia with macrophages containing clusters of numerous yeast cells. Inflammatory lesions and yeast also were found in pulmonary, mediastinal, prescapular, and duodenal lymph nodes, spleen, liver, kidneys, urinary bladder, pancreas, right adrenal gland, and the pyloric stomach. Histomorphology, fungal culture, and polymerase chain reaction analysis indicated that the fungus was Histoplasma capsulatum var. capsulatum. This is the first report of histoplasmosis in a cetacean.


Deficiency of intestinal mucin-2 protects mice from diet-induced fatty liver disease and obesity.
Nonalcoholic fatty liver disease (NAFLD) and obesity are characterized by altered gut microbiota, inflammation, and gut barrier dysfunction. Here, we investigated the role of mucin-2 (Muc2) as the major component of the intestinal mucus layer in the development of fatty liver disease and obesity. We studied experimental fatty liver disease and obesity induced by feeding wild-type and Muc2-knockout mice a high-fat diet (HFD) for 16 wk. Muc2 deficiency protected mice from HFD-induced fatty liver disease and obesity. Compared with wild-type mice, after a 16-wk HFD, Muc2-knockout mice exhibited better glucose homeostasis, reduced inflammation, and upregulated expression of genes involved in lipolysis and fatty acid β-oxidation in white adipose tissue. Compared with wild-type mice that were fed the HFD as well, Muc2-knockout mice also displayed higher intestinal and plasma levels of IL-22 and higher intestinal levels of the IL-22 target genes Reg3b and Reg3g. Our findings indicate that absence of the intestinal mucus layer activates the mucosal immune system. Higher IL-22 levels protect mice from diet-induced features of the metabolic syndrome.


Fifty-year trend in hip fracture incidence.
During the 50-year period 1928 to 1977, 1,250 Rochester, Minnesota residents experienced 1,355 proximal femur (hip) fractures. The incidence of these fractures appeared to rise during the first 15 years of the study, but this was most likely due to underascertainment of cases in the early period since there was no evidence of a cohort effect. From 1943 onward, there was no significant increase in the incidence rates which remained relatively level for total hip fractures, initial hip fractures alone, and for initial hip fractures due to moderate trauma. Thus, we found no evidence to support the contention that hip fracture incidence has risen dramatically in recent years. Secular changes in incidence cannot account for the observation that hip fracture rates in Rochester are greater than those reported from earlier population-based studies.


Avian skin development and the evolutionary origin of feathers.
The discovery of several dinosaurs with filamentous integumentary appendages of different morphologies has stimulated models for the evolutionary origin of feathers. In order to understand these models, knowledge of the development of the avian integument must be put into an evolutionary context. Thus, we present a review of avian scale and feather development, which summarizes the morphogenetic events involved, as well as the expression of the beta (beta) keratin multigene family that characterizes the epidermal appendages of reptiles and birds. First we review information on the evolution of the ectodermal epidermis and its beta (beta) keratins. Then we examine the morphogenesis of scutate scales and feathers including studies in which the extraembryonic ectoderm of the chorion is used to examine dermal induction. We also present studies on the scaleless (sc) mutant, and, because of the recent discovery of "four-winged" dinosaurs, we review earlier studies of a chicken strain, Silkie, that expresses ptilopody (pti), "feathered feet." We conclude that the ability of the ectodermal epidermis to generate discrete cell populations capable of forming functional structural elements consisting of specific members of the beta keratin multigene family was a plesiomorphic feature of the archosaurian ancestor of crocodilians and birds. Evidence suggests that the discrete epidermal lineages that make up the embryonic feather filament of extant birds are homologous with similar embryonic lineages of the developing scutate scales of birds and the scales of alligators. We believe that the early expression of conserved signaling modules in the embryonic skin of the avian ancestor led to the early morphogenesis of the embryonic feather filament, with its periderm, sheath, and barb ridge lineages forming the first protofeather. Invagination of the epidermis of the protofeather led to formation of the follicle providing for feather renewal and diversification. The observations that scale formation in birds involves an inhibition of feather formation coupled with observations on the feathered feet of the scaleless (High-line) and Silkie strains support the view that the ancestor of modern birds may have had feathered hind limbs similar to those recently discovered in nonavian dromaeosaurids. And finally, our recent observation on the bristles of the wild turkey beard raises the possibility that similar integumentary appendages may have adorned nonavian dinosaurs, and thus all filamentous integumentary appendages may not be homologous to modern feathers.


Clinical evaluation of four dental amalgams over a two year period.
The aim of this study was to determine the clinical durability of two locally manufactured amalgams. In each of eighty-two patients one class II cavity was restored with Dispersalloy (Johnson and Johnson) which served as the control and all the other class II cavities were randomly restored with one of the following amalgams: Amalgaphase, Amalga 43 (Amalgam Alloys-South Africa) or Permite C (Southern Dental Industries). Matrix bands were placed. Kalzinol bases and Polyvar varnish were applied in all cavities. All amalgams were mixed according to the manufacturers' instructions. The cavities were overpacked with amalgam, and the amalgam condensed by hand, carved and then burnished with a ball burnisher. Twenty-four hours later all restorations were polished with a Shofu polishing system and colour photographs were taken. The restorations were evaluated using the Ryge and Snyder (1973) evaluation system, as well as comparison of the colour photos. The Fisher's Exact Test was used for the statistical analysis. During the second year there were no significant (p < 0.05) differences between the four brands of amalgam restorations except as regards the gloss category. The two South African manufactured amalgams compared well with the two imported amalgams. During the second year the marginal integrity and surface texture of Amalgaphase compared well with that of Permite C. Dispersalloy and Permite C showed more deterioration in anatomic form than the two South African amalgams. Amalgaphase was the only amalgam to show no bulk fracture within a period of two years. Amalgaphase performed better in the gloss category than the other three amalgams during the second year evaluation. According to the weighted product calculation Amalgaphase was the best amalgam followed by Dispersalloy, Amalga 43 and Permite C.


Review of international safety data for lisinopril-hydrochlorothiazide combination treatment.
A total of 930 patients have been evaluated for safety in a programme of clinical trials for lisinopril-hydrochlorothiazide combination treatment. Combination therapy with these two agents is generally well tolerated. In clinical trials, adverse experiences in patients treated with a lisinopril-hydrochlorothiazide combination were dizziness (7.5%), headache (5.2%), cough (3.9%), fatigue (3.7%), orthostatic effects (3.2%), diarrhoea (2.5%), nausea (2.2%) and upper respiratory tract infection (2.2%). Withdrawals from treatment have been relatively infrequent comprising dizziness (0.8%), headache (0.3%), cough (0.6%), fatigue (0.4%), diarrhoea (0.2%), orthostatic effects and nausea (0.1% each). The most common laboratory adverse experiences in patients on therapy with the lisinopril-hydrochlorothiazide combination are: increases in serum glucose, triglycerides, uric acid, serum creatinine, blood urea nitrogen and blood urea; and decreases in serum potassium. However, in individual controlled studies, the addition of lisinopril to treatment with hydrochlorothiazide results in attenuation of some of the potentially adverse metabolic affects of the diuretic. Adverse experiences in the patients treated for periods of 50 weeks or more, the elderly and the renally impaired are similar to those seen in the total population. Overall the available data indicate that a fixed dose combination of lisinopril-hydrochlorothiazide is a well-tolerated therapeutic option in patients with mild-to-moderate hypertension.


[Osteomatous chronic otitis. 27 new cases. 2 new forms].
The authors report 27 new cases of osteomatous chronic otitis, with (15) or without (12) cholesteatomas, with perforated (13) or intact (14) drum, the latter sometimes having a very atypical appearance (2), frankly purulent or simply inflammatory. This results in a wide range of clinical appearances. There are two macroscopic forms: the diffuse attico-antral form and localized forms (drum - C.S.C.L.). The authors emphasize: the primordial interest of tomography (or CT scan), the keystone of their discovery, as well as the dangers of their surgical treatment. In addition to review of these known concepts, two forms have been identified: 1) Osteomatous attico-chain monoblock, where the ossicles of the ear are adherent in their entirety, either to the external attical wall or, much more rarely, to the medial attical wall. They describe 13 cases, the pattern of which is very different, according to whether the diagnosis is one of cholesteatomatous chronic otitis or not. In the absence of cholesteatoma (8 cases), the clinical picture is suggestive of otospongiosis or of chronic otitis with intact drum. The surgical procedure is related to these findings: 1 completion of myringoplasty, 7 piston-malleus. In cases of cholesteatomatous chronic otitis (5 cases), the picture is completely different, the cholesteatoma developing at leisure, behind the impenetrable osteomatous shield. Once again, the surgical procedure is modified: 4 vast evacuation cavities and one mastoatticotomy for giant cholesteatoma with intact drum. 2) Diffuse and "progressive" osteomatous stenosis of the walls of the E.A.M. The authors describe 6 cases occurring either during cholesteatomatous chronic otitis, or within a few weeks, simulating chronic otitis with intact drum.


Improvements in pincer nail deformity occur earlier and last longer when acetylcysteine gel is added to an overcurvature-correcting device: Results of a phase 3, multicenter, randomized, vehicle-controlled, investigator-blinded study.
Acetylcysteine (AC) destabilizes keratin and softens nails, reducing the time needed to correct pincer nail deformity with an overcurvature-correcting device. The objective of this phase 3, multicenter, randomized, investigator-blinded study was to evaluate the early and sustained therapeutic effectiveness and safety of 10% AC gel plus an overcurvature-correcting device to treat pincer nails. Patients aged 12 years and older with hallux pincer nail were fitted with an overcurvature-correcting device for 7 days, with a 24-h application of AC gel (n = 40) or vehicle (n = 39) on day 1. The primary end point (achievement of a distal narrowed nail width ratio ≥70% on day 8) was met by 47.5% in the 10% AC group and 25.6% in the vehicle group (difference 21.9%; p = 0.0439). Secondary end points showed a greater tendency towards improvement with 10% AC. The nail correction effect was maintained for at least 12 weeks in the majority of AC-treated patients, although the study duration was insufficient to assess the long-term probability of recurrence. No AC-related adverse events were reported. In conclusion, a single application of 10% AC gel combined with short-term device use facilitated earlier correction of pincer nails compared with the device alone, with improvements maintained after device removal.


Proteoglycan IMPG2 Shapes the Interphotoreceptor Matrix and Modulates Vision.
Photoreceptor neurons are surrounded by an extracellular matrix, called the interphotoreceptor matrix (IPM). Activities crucial to vision occur within the IPM, including trafficking of nutrients and metabolites, retinal attachment, and interactions needed for normal outer segment phagocytosis. The IPM includes the following two unique proteoglycans: IPM proteoglycan 1 (IMPG1) and IMPG2. Patients with mutations in IMPG1/IMPG2 develop visual deficits with subretinal material accumulation, highlighting the critical role of the IPM in vision. To determine the role of these proteoglycans in retinal physiology and the pathologic mechanisms that lead to vision loss, we generated mouse models lacking IMPG1/IMPG2. In normal retina, IMPG1 and IMPG2 occupy distinct IPM compartments, represent the main source of chondroitin sulfate and are fundamental for the constitution of the cone-specific glycocalyx stained by the PNA (peanut agglutinin) lectin marker. No evident morphologic or functional deficits were found in mice lacking IMPG1. In the absence of IMPG2, IMPG1 abnormally accumulated at the subretinal space need, likely leading to the formation of subretinal lesions and reduced visual function. Interestingly, mice lacking both IMPG1 and IMPG2, regardless of sex, showed normal retinal structure and function, demonstrating that the aberrant IMPG1 distribution is the main cause of the visual alterations observed in the absence of IMPG2. In conclusion, our results show the dependence of secreted proteoglycans such as IMPG1 on the extracellular environment to properly integrate into the matrix, demonstrate the role of IMPG2 in shaping the IPM, and shed light on the potential mechanisms leading to the development of subretinal lesions and vision loss.SIGNIFICANCE STATEMENT The photoreceptors are specialized neurons that drive phototransduction in the mammalian retina. These cells are organized and surrounded by an extracellular matrix, the interphotoreceptor matrix (IPM). Mutations in IPM proteoglycans are associated with blindness in humans. Our studies show that two specific proteoglycans of the IPM, IPM proteoglycan 1 (IMPG1) and IMPG2, form a dynamic structure with distinct localization and dependency. When IMPG2 is absent, IMPG1 cannot integrate into the IPM, leading to abnormal proteoglycan accumulation and visual deficits. This work adds a new layer of understanding to IPM physiology and describes the pathologic events following deficits in proteoglycans, providing novel possibilities for visual restoration in patients with IMPG-related pathologies.


Measurement of total bilirubin by use of bilirubin oxidase.
We used an enzymatic method for measuring total bilirubin in serum. Results by this method varied linearly with bilirubin concentrations to at least 300 mg/L. The day-to-day precision (CV) of the method ranged from less than 1% to about 11% at bilirubin concentrations of 183 and 12 mg/L, respectively. Commonly used anticoagulants, serum preparation materials, and selected drugs had no effect on the apparent bilirubin concentration, but turbidity caused a slight increase and hemoglobin concentrations of 2 g/L resulted in lower values, by as much as 17 mg/L at a bilirubin concentration of 95 mg/L. Patients' results obtained with this enzymatic method were slightly lower than those obtained with methods based on the Jendrassik-Grof principle. The largest differences, seen in samples with high "direct" bilirubin concentrations, can be decreased by measuring the absorbance at 425 nm instead of at 465 nm as recommended by the supplier of the bilirubin oxidase method.


Chronological development of functional fluorophores for bio-imaging.
Functional fluorophores represent an emerging research field, distinguished by their diverse applications, especially in sensing and cellular imaging. After the discovery of quinine sulfate and subsequent elucidation of the fluorescence mechanism by Sir George Stokes, research in the field of fluorescence gained momentum. Over the past few decades, advancements in sophisticated instruments, including super-resolution microscopy, have further promoted cellular imaging using traditional fluorophores. These advancements include deciphering sensing mechanisms via photochemical reactions and scrutinizing the applications of fluorescent probes that specifically target organelles. This approach elucidates molecular interactions with biomolecules. Despite the abundance of literature illustrating different classes of probe development, a concise summary of newly developed fluorophores remains inadequate. In this review, we systematically summarize the chronological discovery of traditional fluorophores along with new fluorophores. We briefly discuss traditional fluorophores ranging from visible to near-infrared (NIR) in the context of cellular imaging and in vivo imaging. Furthermore, we explore ten new core fluorophores developed between 2007 and 2022, which exhibit advanced optical properties, providing new insights into bioimaging. We illustrate the utilization of new fluorophores in cellular imaging of biomolecules, such as reactive oxygen species (ROS), reactive nitrogen species (RNS), and proteins and microenvironments, especially pH and viscosity. Few of the fluorescent probes provided new insights into disease progression. Furthermore, we speculate on the potential prospects and significant challenges of existing fluorophores and their potential biomedical research applications. By addressing these aspects, we intend to illuminate the compelling advancements in fluorescent probe development and their potential influence across various fields.


Membrane-Bound PenA β-Lactamase of Burkholderia pseudomallei.
Burkholderia pseudomallei is the etiologic agent of melioidosis, a difficult-to-treat disease with diverse clinical manifestations. β-Lactam antibiotics such as ceftazidime are crucial to the success of melioidosis therapy. Ceftazidime-resistant clinical isolates have been described, and the most common mechanism is point mutations affecting expression or critical amino acid residues of the chromosomally encoded class A PenA β-lactamase. We previously showed that PenA was exported via the twin arginine translocase system and associated with the spheroplast fraction. We now show that PenA is a membrane-bound lipoprotein. The protein and accompanying β-lactamase activity are found in the membrane fraction and can be extracted with Triton X-114. Treatment with globomycin of B. pseudomallei cells expressing PenA results in accumulation of the prolipoprotein. Mass spectrometric analysis of extracted membrane proteins reveals a protein peak whose mass is consistent with a triacylated PenA protein. Mutation of a crucial lipobox cysteine at position 23 to a serine residue results in loss of β-lactamase activity and absence of detectable PenAC23S protein. A concomitant isoleucine-to-alanine change at position 20 in the signal peptide processing site in the PenAC23S mutant results in a nonlipidated protein (PenAI20A C23S) that is processed by signal peptidase I and exhibits β-lactamase activity. The resistance profile of a B. pseudomallei strain expressing this protein is indistinguishable from the profile of the isogenic strain expressing wild-type PenA. The data show that PenA membrane association is not required for resistance and must serve another purpose.


The nature of the non-ultrafilterable glycosaminoglycans of normal human urine.
1. The non-ultrafilterable acidic glycosaminoglycans from pooled urine of normal men, aged about 20, were isolated and characterized. The isolation procedure included digestion with sialidase and pronase, and fractionation by stepwise elution from an ECTEOLA-cellulose column. The glycosaminoglycans in each fraction were separated from each other by preparative electrophoresis in sodium barbital buffer and in barium acetate. 2. Approximate relative amounts of the different glycosaminoglycans were: chondroitin sulphate 60%, chondroitin 2%, hyaluronic acid 4%, dermatan sulphate 1%, heparan sulphate 15% and keratan sulphate 18%. Chondroitin sulphate-dermatan sulphate hybrids seemed to occur in trace amounts. 3. Chondroitin sulphate, heparan sulphate and keratan sulphate were heterogeneous with respect to degree of sulphation. Two distinct groups of chondroitin sulphate fractions were found, with sulphate/hexosamine molar ratios of about 0.5 and 1 respectively. The sulphate/hexosamine molar ratios in the heparan sulphate fractions varied from 0.5 to 0.9; the N-sulphate/hexosamine ratio was about 0.5 in all fractions. The sulphate/hexosamine molar ratios in the keratan sulphate fractions varied from 0.2 to 0.7.


Realistic pathologic classification of acute myeloid leukemias.
Most classification systems of acute myeloid leukemia (AML) rely largely on the criteria proposed by the French-American-British (FAB) Cooperative Group. The recently proposed World Health Organization (WHO) classification of neoplastic diseases of the hematopoietic and lymphoid tissues includes a classification of AMLs. The proposed WHO classification of AMLs includes traditional FAB-type categories of disease, as well as additional disease types that correlate with specific cytogenetic findings and AML associated with myelodysplasia. This system includes a large number of disease categories, many of which are of unknown clinical significance, and there seems to be substantial overlap between disease groups in the WHO proposal. Some disease types in the WHO proposal cannot be diagnosed without detailed clinical information, or they are diagnosed only by the cytogenetic findings. In this report, a realistic pathologic classification for AML is proposed that includes disease types that correlate with specific cytogenetic translocations and can be recognized reliably by morphologic evaluation and immunophenotyping and that incorporates the importance of associated myelodysplastic changes. This system would be supported by cytogenetic or molecular genetic studies and could be expanded as new recognizable clinicopathologic entities are described.


Hijacking Factor H for Complement Immune Evasion.
The complement system is an essential player in innate and adaptive immunity. It consists of three pathways (alternative, classical, and lectin) that initiate either spontaneously (alternative) or in response to danger (all pathways). Complement leads to numerous outcomes detrimental to invaders, including direct killing by formation of the pore-forming membrane attack complex, recruitment of immune cells to sites of invasion, facilitation of phagocytosis, and enhancement of cellular immune responses. Pathogens must overcome the complement system to survive in the host. A common strategy used by pathogens to evade complement is hijacking host complement regulators. Complement regulators prevent attack of host cells and include a collection of membrane-bound and fluid phase proteins. Factor H (FH), a fluid phase complement regulatory protein, controls the alternative pathway (AP) both in the fluid phase of the human body and on cell surfaces. In order to prevent complement activation and amplification on host cells and tissues, FH recognizes host cell-specific polyanionic markers in combination with complement C3 fragments. FH suppresses AP complement-mediated attack by accelerating decay of convertases and by helping to inactivate C3 fragments on host cells. Pathogens, most of which do not have polyanionic markers, are not recognized by FH. Numerous pathogens, including certain bacteria, viruses, protozoa, helminths, and fungi, can recruit FH to protect themselves against host-mediated complement attack, using either specific receptors and/or molecular mimicry to appear more like a host cell. This review will explore pathogen complement evasion mechanisms involving FH recruitment with an emphasis on: (a) characterizing the structural properties and expression patterns of pathogen FH binding proteins, as well as other strategies used by pathogens to capture FH; (b) classifying domains of FH important in pathogen interaction; and (c) discussing existing and potential treatment strategies that target FH interactions with pathogens. Overall, many pathogens use FH to avoid complement attack and appreciating the commonalities across these diverse microorganisms deepens the understanding of complement in microbiology.


The quest for a true One Health perspective of brucellosis.
One Health is an interdisciplinary collaboration that aims at mitigating risks to human health arising from microorganisms present in non-human animal species, which have the potential to be transmitted and cause disease in humans. Different degrees of scientific collaboration and sectoral integration are needed for different types of zoonotic diseases, depending on the health and associated economic gains that can be expected from a One Health approach. Indeed, mitigating zoonotic risks related to emerging diseases with pandemic potential is different from mitigating risks related to endemic zoonotic diseases like brucellosis. Likewise, management of brucellosis at the wildlife-livestock interface in wildlife conservation areas is in essence different from mitigating transmission of a given Brucella species within its preferential host species, which in turn is different from mitigating the spillover of a given Brucella species to non-preferential host species, humans included. Brucellosis economic models often oversimplify and/or wrongly assess transmission between reservoir hosts and spillover hosts. Moreover,they may not properly value non-market outcomes, such as avoidance of human disease, consumer confidence and conservation biology issues. As a result, uncertainty is such that the economic predictions of these models can be questionable. Therefore, understanding the infection biology of Brucella species is a prerequisite. This paper reviews and highlights important features of the infection biology of Brucella species and the changing epidemiology of brucellosis that need to be integrated into a true One Health perspective of brucellosis.


Loss of KCNJ10 protein expression abolishes endocochlear potential and causes deafness in Pendred syndrome mouse model.
Pendred syndrome, a common autosomal-recessive disorder characterized by congenital deafness and goiter, is caused by mutations of SLC26A4, which codes for pendrin. We investigated the relationship between pendrin and deafness using mice that have (Slc26a4+/+) or lack a complete Slc26a4 gene (Slc26a4-/-). Expression of pendrin and other proteins was determined by confocal immunocytochemistry. Expression of mRNA was determined by quantitative RT-PCR. The endocochlear potential and the endolymphatic K+ concentration were measured with double-barreled microelectrodes. Currents generated by the stria marginal cells were recorded with a vibrating probe. Tissue masses were evaluated by morphometric distance measurements and pigmentation was quantified by densitometry. Pendrin was found in the cochlea in apical membranes of spiral prominence cells and spindle-shaped cells of stria vascularis, in outer sulcus and root cells. Endolymph volume in Slc26a4-/- mice was increased and tissue masses in areas normally occupied by type I and II fibrocytes were reduced. Slc26a4-/- mice lacked the endocochlear potential, which is generated across the basal cell barrier by the K+ channel KCNJ10 localized in intermediate cells. Stria vascularis was hyperpigmented, suggesting unalleviated free radical damage. The basal cell barrier appeared intact; intermediate cells and KCNJ10 mRNA were present but KCNJ10 protein was absent. Endolymphatic K+ concentrations were normal and membrane proteins necessary for K+ secretion were present, including the K+ channel KCNQ1 and KCNE1, Na+/2Cl-/K+ cotransporter SLC12A2 and the gap junction GJB2. These observations demonstrate that pendrin dysfunction leads to a loss of KCNJ10 protein expression and a loss of the endocochlear potential, which may be the direct cause of deafness in Pendred syndrome.


Ubiquitin is a component of paired helical filaments in Alzheimer's disease.
Paired helical filaments (PHF), which constitute a distinct type of pathological neuronal fiber, are the principal constituent of neurofibrillary tangles that occur in the brain of patients with Alzheimer's disease. Their insolubility in sodium dodecyl sulfate and urea has prevented the analysis of their subunit composition by gel electrophoresis. A monoclonal antibody (DF2) was isolated that specifically labeled PHF at both the light and electron microscopic levels. It labeled a small polypeptide (5 kilodaltons) that was shown to be ubiquitin in immunoblots of the soluble fraction of brain homogenates. To obtain direct evidence that ubiquitin is a component of PHF, PHF were treated with concentrated formic acid and digested with lysylendopeptidase; ubiquitin-derived peptides were then identified by reversed-phase high-performance liquid chromatography. Two fragments in the PHF digest were identified as derived from ubiquitin by protein sequencing. This procedure should make possible definitive identification of other PHF components.


A technique for making high-resolution megapixel mosaic photomicrographs of nematodes.
Multiple images of a whole nematode specimen were taken with a high power oil-immersion objective lens and joined together to form one high-resolution megapixel, mosaic photomicrograph of the entire specimen, with the use of a relatively new mounting technique made with a 4% water agar pad. The agar pad kept the specimen nearly level and lateral, and when amended with 10 mM sodium azide, this mounting technique gradually paralyzed the nematode in a natural pose to enable production of sharp, clear images. The individual photographs were joined together and merged into one very large, seamless image. These montaged images will be useful for teaching because the student has access to a virtual specimen that is mounted in the correct orientation, imaged with a research grade microscope, and preserved in a narcotized, living condition. Such specimen images can also serve as representatives of type and voucher specimens without the deterioration typical of real types. The files can be copied and viewed with a computer almost anywhere and at any time, rather than using a more cumbersome, limiting, and expensive microscope.


Anti-N-Methyl-D-Aspartate Receptor Encephalitis: A Case Study.
Anti-N-methyl-D-aspartate receptor encephalitis is an autoimmune syndrome that presents with personality changes, autonomic dysfunction, and neurologic deterioration. Most patients with this syndrome progress from psychosis to seizure to catatonia, often associated with abnormal movements, autonomic instability, and hypoventilation. First-line treatment constitutes resection of the associated neoplasm, corticosteroids, intravenous immunoglobulin, and plasma exchange. Second-line treatment includes rituximab and cyclophosphamide. A case of confirmed anti-N-methyl-D-aspartate receptor encephalitis is presented that illustrates the diagnostic and treatment challenges associated with this syndrome and underscores the nursing implications of medical management during immunosuppression. This case study recommends surface cooling and a pharmaceutical regimen for management of autonomic storming, which is a hallmark of this disorder.


Five years of data diuresis: what have WEH learned?
This year represents the fifth annual Data Diuresis session of the Water and Electrolyte Homeostasis (WEH) section of the American Physiological Society (APS) at the 2015 Experimental Biology meeting. As opposed to taking a single organ approach to the study of physiology, the WEH section employs an integrative approach to encompass how the different organ systems interact to regulate numerous physiological and pathophysiological processes. The goal of this minireview is to highlight the broad spectrum of research themes that were presented over the first five years of Data Diuresis. Presentation topics include (but are not limited to) oxidative stress, inflammation, obesity, pregnancy, and hypertension spanning the brain, heart and vasculature, and kidney. WEH researchers continue to impact and help drive the direction of physiological research across multiple disciplines, leaving us excited to see what the next five years of Data Diuresis will bring.


Uncovering degrees of workplace bullying: A comparison of baccalaureate nursing students' experiences during clinical placement in Australia and the UK.
Bullying in health workplaces has a negative impact on nurses, their families, multidisciplinary teams, patient care and the profession. This paper compares the experiences of Australian and UK baccalaureate nursing students in relation to bullying and harassment during clinical placement. A secondary analysis was conducted on two primary cross-sectional studies of bullying experiences of Australian and UK nursing students. Data were collected using the Student Experience of Bullying during Clinical Placement (SEBDCP) questionnaire and analysed using descriptive and inferential statistics. The total sample was 833 Australian and 561 UK students. Australian nursing students experienced a higher rate of bullying (50.1%) than UK students (35.5%). Students identified other nurses as the main perpetrators (Aust 53%, UK 68%), although patients were the main source of physical acts of bullying. Few bullied students chose to report the episode/s. The main reason for non-reporting was fear of being victimised. Sadly, some students felt bullying and harassment was 'part of the job'. A culture of bullying in nursing persists internationally. Nursing students are vulnerable and can question their future in the 'caring' profession of nursing after experiencing and/or witnessing bullying during clinical placement. Bullying requires a zero tolerance approach. Education providers must develop clearer policies and implement procedures to protect students - the future nursing workforce.


The copolymeric structure of pig skin dermatan suplhate. Characterization of D-glucuronic acid-containing oligosaccharides isolated after controlled degradation of oxydermatan sulphate.
Selective periodate oxidation of unsubstituted l-iduronic acid residues in copolymeric dermatan sulphate chains was followed by reduction-hydrolysis or alkaline elimination. By this procedure the glucuronic acid-containing periods were isolated in oligosaccharide form; general formula: [Formula: see text] Further degradation of these oligosaccharides with chondroitinase-AC yielded three types of products: (a) sulphated trisaccharide containing an unsaturated uronosyl moiety in the non-reducing terminal and a C(4) fragment in the reducing terminal, DeltaUA-GalNAc-(-SO(4))-R; (b) monosulphated, unsaturated disaccharide, DeltaUA-GalNAc-SO(4) when n is greater than or equal to 2; and (c) N-acetylgalactosamine with or without sulphate. Oligosaccharides containing a single glucuronic acid residue (n=1) comprised more than half of the glucuronic acid-containing oligosaccharides. The terminal N-acetylgalactosamine moiety of the shortest oligosaccharide was largely 4-sulphated, whereas higher oligosaccharides primarily contained 6-sulphated or unsulphated hexosamine moieties in the same position. Moreover, IdUA-SO(4)-containing oligosaccharides were encountered. These oligosaccharides were resistant to the action of chondroitinase-ABC.


Calibrating divergence times on species trees versus gene trees: implications for speciation history of Aphelocoma jays.
Estimates of the timing of divergence are central to testing the underlying causes of speciation. Relaxed molecular clocks and fossil calibration have improved these estimates; however, these advances are implemented in the context of gene trees, which can overestimate divergence times. Here we couple recent innovations for dating speciation events with the analytical power of species trees, where multilocus data are considered in a coalescent context. Divergence times are estimated in the bird genus Aphelocoma to test whether speciation in these jays coincided with mountain uplift or glacial cycles. Gene trees and species trees show general agreement that diversification began in the Miocene amid mountain uplift. However, dates from the multilocus species tree are more recent, occurring predominately in the Pleistocene, consistent with theory that divergence times can be significantly overestimated with gene-tree based approaches that do not correct for genetic divergence that predates speciation. In addition to coalescent stochasticity, Haldane's rule could account for some differences in timing estimates between mitochondrial DNA and nuclear genes. By incorporating a fossil calibration applied to the species tree, in addition to the process of gene lineage coalescence, the present approach provides a more biologically realistic framework for dating speciation events, and hence for testing the links between diversification and specific biogeographic and geologic events.


Stable isotope-resolved metabolomic analysis of lithium effects on glial-neuronal metabolism and interactions.
Despite the long-established therapeutic efficacy of lithium in the treatment of bipolar disorder (BPD), its molecular mechanism of action remains elusive. Newly developed stable isotope-resolved metabolomics (SIRM) is a powerful approach that can be used to elucidate systematically how lithium impacts glial and neuronal metabolic pathways and activities, leading ultimately to deciphering its molecular mechanism of action. The effect of lithium on the metabolism of three different (13)C-labeled precursors ([U-(13)C]-glucose, (13)C-3-lactate or (13)C-2,3-alanine) was analyzed in cultured rat astrocytes and neurons by nuclear magnetic resonance (NMR) spectroscopy and gas chromatography mass spectrometry (GC-MS). Using [U-(13)C]-glucose, lithium was shown to enhance glycolytic activity and part of the Krebs cycle activity in both astrocytes and neurons, particularly the anaplerotic pyruvate carboxylation (PC). The PC pathway was previously thought to be active in astrocytes but absent in neurons. Lithium also stimulated the extracellular release of (13)C labeled-lactate, -alanine (Ala), -citrate, and -glutamine (Gln) by astrocytes. Interrogation of neuronal pathways using (13)C-3-lactate or (13)C-2,3-Ala as tracers indicated a high capacity of neurons to utilize lactate and Ala in the Krebs cycle, particularly in the production of labeled Asp and Glu via PC and normal cycle activity. Prolonged lithium treatment enhanced lactate metabolism via PC but inhibited lactate oxidation via the normal Krebs cycle in neurons. Such lithium modulation of glycolytic, PC and Krebs cycle activity in astrocytes and neurons as well as release of fuel substrates by astrocytes should help replenish Krebs cycle substrates for Glu synthesis while meeting neuronal demands for energy. Further investigations into the molecular regulation of these metabolic traits should provide new insights into the pathophysiology of mood disorders and early diagnostic markers, as well as new target(s) for effective therapies.


[War and oranges: a radiophonic lecture on the nutritional value of Brazilian fruits (1940)].
Since the early twentieth-century, sanitarians have relied on the radio for educational purposes. Men like Geraldo Horácio de Paula Souza and Borges Vieira used radio waves to popularize hygienic habits and disseminate information on sanitary topics. On April 16, 1940, Paula Souza stood before the microphones at Rádio Educadora Paulista and gave a lecture on the nutritional value of oranges and the need to eat them as a patriotic act. Touting the fruit's nutritional qualities, Paula Souza launched the 'orange campaign'. Its success reached beyond the borders of São Paulo state, as the campaign found space in the press and took form as an advertising brochure published by the Agricultural Information Service, with the unconditional support of the Minister of Agriculture, Fernando Costa.


Ectopic expression of MELK in oral squamous cell carcinoma and its correlation with epithelial mesenchymal transition.
Epithelial-mesenchymal transition (EMT) is closely correlated to metastasis formation generation and maintenance of cancer stem cells, nevertheless, the underlying mechanisms are unclear. The aim of this study is to investigate the role of maternal embryonic leucine-zipper kinase (MELK) in EMT regulation in oral squamous cell carcinoma (OSCC). We found that there was overexpression of MELK in human OSCC tissues, and high MELK expression was correlated with lymphatic metastasis and led to poor prognosis in patients with OSCC. We also confirmed that MELK is closely correlated to the EMT process using a human OSCC tissue microarray. Additionally, MELK expression was observed to be regulated in several OSCC cell lines, and knockdown of MELK genes inhibited cell proliferation, migration, invasion and EMT of OSCC cells in vitro. Furthermore, silencing of MELK suppressed tumour growth in vivo, and experimental research verified that MELK may augment OSCC development via mediating the Wnt/Notch signalling pathway. Our findings suggest that MELK serves as an oncogene to improve malignant development of OSCC via enhancing EMT, and MELK might be a potential target for anticancer therapeutic.


New developments in the neurobiology of the tuberous sclerosis complex.
To outline recent developments in the neurobiology of the tuberous sclerosis complex (TSC). TSC may be associated with neuropsychiatric disorders including epilepsy, mental retardation, and autism. The uncontrolled growth of subependymal giant cell astrocytomas may lead to hydrocephalus and death. The recent identification of mutations in two genes (TSC1 and TSC2) that cause TSC has led to rapid progress in understanding the molecular and cellular pathogenesis of this disorder. How distinct mutations lead to the varied clinical phenotype of TSC is under intense investigation. We report the recent diagnostic criteria for TSC and provide an overview of the molecular genetics, molecular pathophysiology, and neuropathology of TSC. Important diagnostic criteria for TSC include facial angiofibromas, ungual fibromas, retinal hamartomas, and cortical tubers. Both familial and sporadic TSC cases occur. Approximately 50% of TSC families show genetic linkage to TSC1 and 50% to TSC2. Among sporadic TSC cases, mutations in TSC2 are more frequent and often accompanied by more severe neurologic deficits. Multiple mutational subtypes have been identified in the TSC1 and TSC2 genes. The TSC1 (chromosome 9) and TSC2 (chromosome 16) genes encode distinct proteins, hamartin and tuberin, respectively, which are widely expressed in the brain and may interact as part of a cascade pathway that modulates cellular differentiation, tumor suppression, and intracellular signaling. Tuberin has a GTPase activating protein-related domain that may contribute to a role in cell cycle passage and intracellular vesicular trafficking. Identification of tuberous sclerosis complex (TSC) gene mutations has fostered understanding of how brain lesions in TSC are formed. Further characterization of the roles of hamartin and tuberin will provide potential therapeutic avenues to treat seizures, mental retardation, and tumor growth in TSC.


An overview of salt intake reduction efforts in the Gulf Cooperation Council countries.
Globally, morbidity and mortality from non-communicable diseases (NCDs) are increasing steadily and at an alarming rate. High blood pressure is a major risk factor for cardiovascular disease (CVD) and salt reduction is an effective measure to decrease mortality rates. In the Eastern Mediterranean region, current salt intake is high, with an average intake of >12 g per person per day. Reducing the intake of salt has been identified as a priority intervention to reduce NCDs. Countries of the Gulf Cooperation Council (GCC) are showing a willingness to comply with the World Health Organization (WHO) recommendations and an eagerness to reduce the burden of NCDs. However, they face some challenges, including lack of political commitment, lack of experience, and shortage of qualified human resources. Salt intake reduction efforts vary in the GCC region, from achieving 20% salt reduction in bread, to the very early stages of planning.


Reduced Lateral Root Branching Density Improves Drought Tolerance in Maize.
An emerging paradigm is that root traits that reduce the metabolic costs of soil exploration improve the acquisition of limiting soil resources. Here, we test the hypothesis that reduced lateral root branching density will improve drought tolerance in maize (Zea mays) by reducing the metabolic costs of soil exploration, permitting greater axial root elongation, greater rooting depth, and thereby greater water acquisition from drying soil. Maize recombinant inbred lines with contrasting lateral root number and length (few but long [FL] and many but short [MS]) were grown under water stress in greenhouse mesocosms, in field rainout shelters, and in a second field environment with natural drought. Under water stress in mesocosms, lines with the FL phenotype had substantially less lateral root respiration per unit of axial root length, deeper rooting, greater leaf relative water content, greater stomatal conductance, and 50% greater shoot biomass than lines with the MS phenotype. Under water stress in the two field sites, lines with the FL phenotype had deeper rooting, much lighter stem water isotopic signature, signifying deeper water capture, 51% to 67% greater shoot biomass at flowering, and 144% greater yield than lines with the MS phenotype. These results entirely support the hypothesis that reduced lateral root branching density improves drought tolerance. The FL lateral root phenotype merits consideration as a selection target to improve the drought tolerance of maize and possibly other cereal crops.


A genetic strategy generating wheat with very high amylose content.
Resistant starch (RS), a type of dietary fibre, plays an important role in human health; however, the content of RS in most modern processed starchy foods is low. Cereal starch, when structurally manipulated through a modified starch biosynthetic pathway to greatly increase the amylose content, could be an important food source of RS. Transgenic studies have previously revealed the requirement of simultaneous down-regulation of two starch branching enzyme (SBE) II isoforms both located on the long arm of chromosome 2, namely SBEIIa and SBEIIb, to elevate the amylose content in wheat from ~25% to ~75%. The current study revealed close proximity of genes encoding SBEIIa and SBEIIb isoforms in wheat with a genetic distance of 0.5 cM on chromosome 2B. A series of deletion and single nucleotide polymorphism (SNP) loss of function alleles in SBEIIa, SBEIIb or both was isolated from two different wheat populations. A breeding strategy to combine deletions and SNPs generated wheat genotypes with altered expression levels of SBEIIa and SBEIIb, elevating the amylose content to an unprecedented ~85%, with a marked concomitant increase in RS content. Biochemical assays were used to confirm the complete absence in the grain of expression of SBEIIa from all three genomes in combination with the absence of SBEIIb from one of the genomes.


Fluorescent Hybridization of Mycobacterium leprae in Skin Samples Collected in Burkina Faso.
Leprosy is caused by Mycobacterium leprae, and it remains underdiagnosed in Burkina Faso. We investigated the use of fluorescent in situ hybridization (FISH) for detecting M. leprae in 27 skin samples (skin biopsy samples, slit skin samples, and skin lesion swabs) collected from 21 patients from Burkina Faso and three from Côte d'Ivoire who were suspected of having cutaneous leprosy. In all seven Ziehl-Neelsen-positive skin samples (four skin biopsy samples and three skin swabs collected from the same patient), FISH specifically identified M. leprae, including one FISH-positive skin biopsy sample that remained negative after testing with PCR targeting the rpoB gene and with the GenoType LepraeDR assay. Twenty other skin samples and three negative controls all remained negative for Ziehl-Neelsen staining, FISH, and rpoB PCR. These data indicate the usefulness of a microscopic examination of skin samples after FISH for first-line diagnosis of cutaneous leprosy. Accordingly, FISH represents a potentially useful point-of-care test for the diagnosis of cutaneous leprosy.


The sensitivity and specificity of the routine kidney biopsy immunofluorescence panel are inferior to diagnosing renal immunoglobulin-derived amyloidosis by mass spectrometry.
Immunoglobulin light chain amyloidosis is the most frequent type of renal amyloidosis in the United States, accounting for 81% of cases. Accurate typing is crucial for early diagnosis and treatment of immunoglobulin-derived amyloidosis and to avoid treating other amyloidoses with potentially toxic chemotherapy. Immunofluorescence is the first step to type renal immunoglobulin-derived amyloidosis but the performance characteristics of this method are largely unknown. Here, we establish the sensitivity and specificity of immunofluorescence for diagnosing immunoglobulin-derived amyloidosis in patients whose amyloid typing was performed by the current gold standard of laser microdissection/mass spectrometry. Renal biopsy pathology reports originating from several institutions with a diagnosis of amyloidosis and which had amyloid typing by laser microdissection/mass spectrometry performed at our center were reviewed. Reported immunofluorescence staining for kappa or lambda of 2+ or more, with weak or no staining for the other light chain was considered positive for light chain amyloidosis by immunofluorescence. Based on microdissection/mass spectrometry results, of the 170 cases reviewed, 104 cases were typed as immunoglobulin-derived amyloidosis and 66 were typed as non-immunoglobulin-derived amyloidosis. Immunofluorescence sensitivity for diagnosing immunoglobulin-derived amyloidosis was 84.6%. The remaining 16 cases could not be diagnosed by immunofluorescence due to reported weak staining for all antigens or reported lack of preferential staining for one antigen. Immunofluorescence specificity was 92.4%. Five cases, all amyloid A amyloidosis, were misdiagnosed as immunoglobulin-derived amyloidosis by immunofluorescence. Immunofluorescence failed to accurately differentiate immunoglobulin-derived from non-immunoglobulin-derived amyloidosis in 12.3% of cases of renal amyloidosis. Relying on immunofluorescence alone for determining immunoglobulin-derived vs. non-immunoglobulin-derived amyloidosis may lead to misdiagnosis. Thus, immunofluorescence has inferior sensitivity and specificity compared with laser microdissection/mass spectrometry in the typing of immunoglobulin-derived amyloidosis.


A critique of the regulation of data science in healthcare research in the European Union.
The EU offers a suitable milieu for the comparison and harmonisation of healthcare across different languages, cultures, and jurisdictions (albeit with a supranational legal framework), which could provide improvements in healthcare standards across the bloc. There are specific ethico-legal issues with the use of data in healthcare research that mandate a different approach from other forms of research. The use of healthcare data over a long period of time is similar to the use of tissue in biobanks. There is a low risk to subjects but it is impossible to gain specific informed consent given the future possibilities for research. Large amounts of data on a subject present a finite risk of re-identification. Consequently, there is a balancing act between this risk and retaining sufficient utility of the data. Anonymising methods need to take into account the circumstances of data sharing to enable an appropriate balance in all cases. There are ethical and policy advantages to exceeding the legal requirements and thereby securing the social licence for research. This process would require the examination and comparison of data protection laws across the trading bloc to produce an ethico-legal framework compatible with the requirements of all member states. Seven EU jurisdictions are given consideration in this critique.


Regional Increase in the Expression of the BCAT Proteins in Alzheimer's Disease Brain: Implications in Glutamate Toxicity.
The human branched chain aminotransferases (hBCATm, mitochondrial and hBCATc, cytosolic) are major contributors to brain glutamate production. This excitatory neurotransmitter is thought to contribute to neurotoxicity in neurodegenerative conditions such as Alzheimer's disease (AD) but the expression of hBCAT in this disease has not previously been investigated. The objective of investigating hBCAT expression is to gain insight into potential metabolic pathways that may be dysregulated in AD brain, which would contribute to glutamate toxicity. Western blot analysis and immunohistochemistry were used to determine the expression and localization of hBCAT in postmortem frontal and temporal cortex from AD and matched control brains. Western blot analysis demonstrated a significant regional increase in hBCATc expression in the hippocampus (↑ 36%; p-values of 0.012), with an increase of ↑ 160% reported for hBCATm in the frontal and temporal cortex (p-values = 4.22 × 10⁻⁴ and 2.79 × 10⁻⁵, respectively) in AD relative to matched controls, with evidence of post-translational modifications to hBCATm, more prominent in AD samples. Using immunohistochemistry, a significant increase in immunopositive labelling of hBCATc was observed in the CA1 and CA4 region of the hippocampus (p-values = 0.011 and 0.026, respectively) correlating with western blot analysis. Moreover, the level of hBCATm in the frontal and temporal cortex correlated significantly with disease severity, as indicated by Braak staging (p-values = 5.63 × 10⁻⁶ and 9.29 × 10⁻⁵, respectively). The expression of the hBCAT proteins is significantly elevated in AD brain. This may modulate glutamate production and toxicity, and thereby play a role in the pathogenesis of the disease.


Beneficial effect of a thromboxane synthetase inhibitor in traumatic shock.
Traumatic shock was induced in anesthetized rats using the Noble-Collip method. This resulted in an abrupt decline in mean arterial blood pressure (MABP) and heart rate. Plasma cathepsin D activity increased sixfold, plasma thromboxane B2 (TxB2) concentration increased 2.5-fold, plasma myocardial depressant factor (MDF) activity increased 3.5 fold, and the mean survival time was 1.4 +/- 0.2 hours. Administration of the selective thromboxane synthetase inhibitor 5-(3-pyridinylmethyl) benzofuran-2-carboxylate (U-63,557A) (4 mg/kg) resulted in a significant improvement in survival time, 3.3 +/- 0.5, p less than 0.01. Plasma cathepsin D activity was not affected by U-63,557A (7.4 +/- 0.8 vs. 8.5 +/- 1.1 U/ml). However, both plasma and peritoneal fluid TxB2 concentration were significantly reduced and accumulation of the toxic peptide, MDF, was significantly blunted (69 +/- 6 vs. 40 +/- 5 U/ml, p less than 0.01). Our data indicate that blockade of thromboxane A2 (TxA2) production by selective synthetase inhibition is beneficial in trauma and support a role for TxA2 in the pathogenesis of circulatory shock.


Ecological rationality or nested sets? Individual differences in cognitive processing predict Bayesian reasoning.
The presentation of a Bayesian inference problem in terms of natural frequencies rather than probabilities has been shown to enhance performance. The effect of individual differences in cognitive processing on Bayesian reasoning has rarely been studied, despite enabling us to test process-oriented variants of the two main accounts of the facilitative effect of natural frequencies: The ecological rationality account (ERA), which postulates an evolutionarily shaped ease of natural frequency automatic processing, and the nested sets account (NSA), which posits analytical processing of nested sets. In two experiments, we found that cognitive reflection abilities predicted normative performance equally well in tasks featuring whole and arbitrarily parsed objects (Experiment 1) and that cognitive abilities and thinking dispositions (analytical vs. intuitive) predicted performance with single-event probabilities, as well as natural frequencies (Experiment 2). Since these individual differences indicate that analytical processing improves Bayesian reasoning, our findings provide stronger support for the NSA than for the ERA.