A comparative UPLC-Q-TOF/MS-based metabolomics approach for distinguishing peach (Prunus persica (L.) Batsch) fruit cultivars with varying antioxidant activity.
Peaches (Prunus persica (L.) Batsch) are nutritionally and economically important and they are one of the most popular fruits consumed worldwide. Understanding metabolite-caused bioactivity differences among cultivars is essential for designing a peach with enhanced nutritional traits. In this study, we report an untargeted UPLC-Q-TOF/MS-based metabolomics approach for comprehensively discriminating between peaches with different antioxidant activities. Mature fruit from 40 peach cultivars were distinguished using principal component analysis and orthogonal partial least squares discrimination analysis. Seventeen differential metabolites were tentatively identified between peach cultivars with high and low antioxidant potency composite indices, and eight metabolites, including procyanidin C1, procyanidin trimer isomer 1, procyanidin trimer isomer 2, procyanidin B1, procyanidin B2, procyanidin B3, prunus inhibitor b, and phloridzin, were identified as marker compounds responsible for the discrimination of the cultivars base on potential antioxidant activity. Our study highlights the essence and predictive power of metabolomics for detecting small differences and for identifying potential marker metabolites based on their levels and composition in plants exhibiting varying bioactivities. Overall, the variations in the metabolites in peach pulp reflected the diversity in the peach germplasm, and these eight compounds are good candidate markers for future genetic breeding of peach fruit with enhanced antioxidant activity.


Impairment on sperm quality and fertility of adult rats after antiandrogen exposure during prepuberty.
This study evaluated the effects of antiandrogen exposure during the prepubertal period on reproductive development and reproductive competence in adults. Male rats were divided into two groups: flutamide, receiving 25 mg/kg/day of flutamide by oral gavage and control, receiving vehicle daily. Dosing continued from PND 21 to 44, and animals were killed on PND 50 or PND 75-80. The epididymis, prostate, vas deferens and seminal vesicle weights were lower in Flutamide group on PND 50, while on PND 80 only seminal vesicle weight was reduced. Fertility assessed by IUI revealed a decrease in the fertility potential in the flutamide-treated adults. Flutamide accelerated sperm transit time through the epididymis, impairing sperm motility and storage. A quantitative analysis of the cauda sperm membrane proteome revealed a few significant changes in protein expression. Thus, exposure to flutamide during the prepubertal period compromises the function of the epididymis along with epididymal sperm quality at adulthood.


Men's use of positive strategies for preventing and managing depression: A qualitative investigation.
One in eight men experience depression and men account for 75% of suicides. Previous research has focused on men's reluctance to seek help and use of unhelpful coping strategies. Thematic analysis was used on transcripts from 21 focus groups and 24 in-depth interviews focused on positive strategies men use to prevent and manage depression. In total, 168 men were recruited and the majority (63%) reported no current depression. Four major themes were identified, where men: (1) used a broad variety of positive strategies and made clear distinctions between prevention and management, (2) used strategies that were "typically masculine", as well as challenged expectations of manliness, (3) felt powerless in the face of suicide, and (4) had accumulated wisdom they felt was beneficial for others. Men specifically advised others to talk about problems. Prevention relied upon regular routines for "balance", while management relied upon "having a plan". The majority of the men were aged over 55 years and highly educated. Younger men or those without tertiary education may favour different strategies. In contrast to using only unhelpful strategies, the men used a broad range of positive strategies and adapted their use depending on mood, symptom or problem severity. Use of positive strategies was sophisticated, nuanced, and often underlined by a guiding philosophy. Rather than simply reacting to problems, men actively engaged in preventing the development of depressed moods, and made conscious choices about when or how to take action. Clinical and public health implications are discussed.


The use of micropatterning to control smooth muscle myosin heavy chain expression and limit the response to transforming growth factor β1 in vascular smooth muscle cells.
In the healthy artery, contractile vascular smooth muscle cells (VSMCs) have an elongated shape and are highly aligned but transition to a synthetic phenotype in culture, while additionally becoming well spread and randomly organized. Thus, controlling VSMC phenotype is a challenge in tissue engineering. In this study, we investigated the effects of micropatterning on contractile protein expression in VSMCs at low and high passage and in the presence of transforming growth factor beta 1 (TGFβ1). Micropatterning led to significantly decreased cell area, increased elongation, and increased alignment compared to non-patterned VSMCs independent of passage number. In the presence of serum, micropatterning led to increased smooth muscle myosin heavy chain (SM-MHC) and α-actin expression in low passage VSMCs, but had no effect on high passage VSMCs. Micropatterning was as effective as TGFβ1 in up-regulating SM-MHC at low passage; however, micropatterning limited VSMC response to TGFβ1 at both low and high passage. Investigation of TGFβ receptor 1 revealed higher expression in non-patterned VSMCs compared to patterned at high passage. Our studies demonstrate that micropatterning is an important regulator of SM-MHC expression in contractile VSMCs and that it may provide a mechanism for phenotype stabilization in the presence of growth factors.


Gonadotrophin secretion in the second half of the menstrual cycle: a comparison of women with normal cycles, luteal phase defects and disturbed follicular development.
In order to study the relationship between episodic gonadotrophin secretion and alterations of ovarian hormone secretion, we examined women with normal menstrual cycles (n = 26), luteal phase defects (n = 10) or disturbed follicular oestradiol secretion (n = 8) as established by daily (except weekends) determinations of oestradiol and progesterone. Pulsatile gonadotrophin secretion was studied during the luteal phase or the second half of the menstrual cycle sampling at 15 min intervals for 12 h. LH and FSH mean concentrations and LH pulse frequency were significantly (P less than 0.01) increased in the group with disturbed follicular development in the presence of decreased oestradiol (E2) and progesterone (P4) levels. In women with luteal phase defects mean LH and FSH concentrations and pulsatile LH secretion showed a nonsignificant trend to lower values in the presence of significantly decreased P4 concentrations during the luteal phase.


Maternal acute fatty liver of pregnancy and the associated risk for long-chain 3-hydroxyacyl-coenzyme a dehydrogenase (LCHAD) deficiency in infants.
Acute fatty liver disease of pregnancy has been recognized as a clinical problem since the 1980s. In the past 8 years, the association of this disease with a genetic inborn error of metabolism in the infant has been recognized. Women who are heterozygous for this disorder are usually asymptomatic until the capacity of their livers to metabolize free fatty acids (FFA) is overwhelmed by a homozygous fetus. The inborn error of metabolism, long-chain 3-hydroxyacyl-coenzyme A dehydrogenase (LCHAD) deficiency, may not be immediately recognizable in the infant. Symptoms in the infant are often triggered by an increased long-chain fatty acid load in the diet, or by illness that results in breakdown of endogenous fat. The following case study reviews the clinical pathophysiology of this perinatal health problem and highlights the priorities for the care of infants born to mothers with acute fatty liver disease of pregnancy.


Characteristics of pulsed electric field cardiac ablation porcine treatment zones with a focal catheter.
Pulsed electric field (PEF) therapies employ punctuated energy delivery to kill cells in a volume of tissue through mechanisms that are not dependent on thermal processes. A key component to successful cardiac ablation procedures is ensuring the generation of transmural, contiguous ablation zones, which requires in-depth knowledge regarding treatment sizes for a given therapeutic application. In this study, a series of acute treatments were delivered to porcine ventricles, where triphenyl tetrazolium chloride (TTC) vitality stain was used to identify treatment effect sizes for the three focal monopolar CENTAURI PEF cardiac ablation energy settings. Treatment depths were 5.7, 7.2, and 8.2 mm for the 19, 22, and 25 A energy settings, respectively. Gross pathology indicated umbral zones of hemorrhage surrounded by pale avital TTC-negative-negative tissue, which contrasted significantly from radiofrequency ablation (RF) controls. Histologically, treatment zones are identified by regions of contraction band necrosis and cardiomyocytolysis, which contrasted with RF control lesions composed primarily of coagulation necrosis. Together, these data indicate the ability for focal monopolar PEF treatments to generate deep treatment zones in cardiac ablation without incurring the gross or histological coagulative characteristics of RF thermal lesions.


Pilot randomized trial of bupropion for adolescent methamphetamine abuse/dependence.
To perform a pilot clinical trial of bupropion for methamphetamine abuse/dependence among adolescents. Nineteen adolescents with methamphetamine abuse (n = 2) or dependence (n = 17) were randomly assigned to bupropion SR 150 mg twice daily or placebo for 8 weeks with outpatient substance abuse counseling. Bupropion was well-tolerated except for one female in the bupropion group who was hospitalized for suicidal ideation during a methamphetamine relapse. Adolescents receiving bupropion and females provided significantly fewer methamphetamine-free urine tests compared to participants receiving placebo (p = .043) and males (p = .005) respectively. Results do not support the feasibility of additional trials of bupropion for adolescent methamphetamine abuse/dependence. Future studies should investigate the influence of gender on adolescent methamphetamine abuse and treatment outcomes.


Tear osmolarity in unilateral pseudoexfoliation syndrome.
The aim was to evaluate tear osmolarity of patients with clinically unilateral pseudoexfoliation (PEX) syndrome and to compare the values with otherwise normal subjects. Sixty-two eyes of 31 patients with unilateral PEX syndrome and 31 eyes of 31 normal subjects were studied. The TearLab osmolarity system (OcuSense, Inc, San Diego, CA, USA) was used to measure tear osmolarity. Eyes were grouped as normal eyes of patients having unilateral PEX syndrome (group A), eyes of patients having unilateral PEX syndrome with deposits of PEX material (group B) and eyes of normal subjects (group C). Differences in tear osmolarity were compared among groups A, B and C. Independent t-tests and paired t-tests were used for statistical analysis. The mean tear osmolarity was 298.7 ± 7.8 mOsm/l (range 285 to 306) in the control group (group C), 306.3 ± 6.6 mOsm/l (range 290 to 314) in the eyes of patients having PEX syndrome with deposits of pseudoexfoliative material (group B) and 301.4 ± 7.1 mOsm/l (range 284 to 305) in the fellow eye of these patients (group A). There was a significant difference between tear osmolarity of groups C and B (independent t-test, p = 0.011). Also, there was a significant difference between the osmolarity of each eye of patients with unilateral PEX syndrome (groups A and B) (paired t-test, p = 0.001). The difference between tear osmolarity of groups A and C was not statistically significant (independent t-test, p = 0.146). In conclusion, tear osmolarity is higher in both eyes of patients when compared with normal subjects, using the independent t-test (although highest in clinically positive eyes of these patients). Both eyes of patients having PEX syndrome could be more prone to the development of dry eye syndrome.


Optokinetic nystagmus correlates with severity of vection-induced motion sickness and gastric tachyarrhythmia.
The present study investigated the relationship of the frequency of nystagmus to the severity of optokinetic rotation-induced motion sickness. Eighty-seven subjects viewed a vertically striped, rotating drum for 16 min. Subjects' electrooculograms, ratings of vection, and subjective symptoms of motion sickness (SSMS) were measured during the drum rotation period. Subjects' electrogastrograms (EGGs) were recorded throughout the baseline and drum rotation periods. The ratios of the spectral intensity of EGG 4-9 cycles per minute (cpm) between drum rotation and baseline periods were calculated. The results indicated that the frequency of nystagmus was positively correlated with the ratings of vection (r = 0.40, p < 0.0001), scores of SSMS (r = 0.61, p < 0.0001), and ratios of EGG 4-9 cpm spectral intensity between drum rotation and baseline periods (r = 0.54, p < 0.0001). These results demonstrated that more rapid eye movement indexed by higher frequency of optokinetic nystagmus are related to the development of symptoms of motion sickness and gastric tachyarrhythmia.


Governance, expertise, and the 'culture of care': The changing constitutions of laboratory animal research in Britain, 1876-2000.
This article examines why early twenty-first century animal research governance in Britain foregrounds the 'culture of care' as its key problem. It adopts a historical perspective to understand why the regulation of animal research became primarily a problem of 'culture', a term firmly associated with the social relations of animal research, at this time and not before. Drawing on the theoretical insights of Sheila Jasanoff, Stephen Hilgartner and others, we contrast the British regulatory framework under the Cruelty to Animals Act (1876), which established statutory regulation of animal research for the first time in the world, with its successor the Animals (Scientific Procedures) Act 1986 (ASPA), in an attempt to chart two closely related yet distinct 'constitutions' of animal research each shaped by a historically situated sociotechnical imaginary. Across this longue durée, many concerns remained consistent yet inevitably, as the biomedical sciences transformed in scale and scope, new concerns emerged. Animal care, at least as far as it entailed a commitment to the prevention of animal suffering, was a prominent feature of animal research governance across the period. However, a concern for the culture and social relations of animal research emerged only in the latter half of the twentieth century. We account for this change primarily through a gradual distribution of responsibility for animal research from a single coherent community with broadly shared expertise ('scientists' with experience of animal research) to a diversified community of multiple experience and skillsets which included, importantly, a more equitable inclusion of animal welfare as a form of expertise with direct relevance to animal research. We conclude that animal research governance could only become conceived as a problem of 'culture' and thus social relations when responsibility for care and animal welfare was distributed across a differentiated community, in which diverse forms of expertise were required for the practice of humane animal research.


All-Cause Mortality in Reproductive-Aged Females by State: An Analysis of the Effects of Abortion Legislation.
To estimate the association between state-level abortion legislation and all-cause mortality among all females of reproductive age and maternal, fetal, and infant mortality. We conducted a retrospective cohort study using the Centers for Disease Control and Prevention's WONDER (Wide-ranging ONline Data for Epidemiologic Research) database. Generalized estimating equations were used to estimate the association between supportive, moderate, and restrictive state abortion regulations and all-cause mortality in reproductive-aged females. Secondary outcomes included maternal, fetal, and infant mortality. The association of the number and type of laws on mortality were estimated. Moderate and supportive states were not associated with a significant decrease in all-cause mortality compared with restrictive states. Maternal mortality (per 100,000 live births) was significantly lower in moderate (-5.79, 95% CI -9.88 to -1.70) compared with restrictive states, but not supportive states (-2.51, 95% CI -6.75 to 1.72). Infant mortality (per 1,000 live births) was significantly lower in both moderate (-0.56, 95% CI -1.09 to -0.04) and supportive (-1.10, 95% CI -1.56 to -0.64) states. Fetal mortality was lower in moderate states (-0.69, 95% CI -1.18 to -0.20) but not in supportive states (-0.64, 95% CI -1.14 to 0.13). Each additional abortion regulation was associated with an increase in maternal mortality (1.09/100,000 live births, 95% CI 0.36-1.82) and infant mortality (0.20/1,000 live births, 95% CI 0.12-0.26). Moderate state abortion legislation was associated with lower rates of maternal, fetal, and infant mortality but not lower all-cause mortality in reproductive-aged females compared with restrictive laws. An increasing number of laws restricting abortion was associated with increased maternal and infant mortality.


Effect of the antiepileptic di-n-propylacetamide on 5-hydroxytryptamine turnover in the brain and 5-hydroxyindoleacetic acid level in the cerebrospinal fluid.
The effect of antiepileptic drug di-n-propylacetamide (DPM) on 5-hydroxytryptamine (5-HT) turnover in rat brain and 5-hydroxyindoleacetic acid (5-HIAA) in cat cerebrospinal fluid (CSF) was investigated. DPM (200 mg/kg) increased brain 5-HIAA without altering the 5-HT level. DPM augmented the accumulation of 5-HT induced by monoamine oxidase inhibition with pargyline (80 mg/kg) and enhanced the accumulation of 5-HIAA in the brain following blockade of transport of this metabolite by probenecid (200 mg/kg). Prior inhibition of 5-HT synthesis by p-chlorophenylalanine (300 mg/kg) abolished the DPM-induced increase in cerebral 5-HIAA. DPM (100 mg/kg) given daily for 5 days considerably elevated 5-HIAA in the CSF of cat during the treatment period. We conclude that DPM increases the turnover of 5-HT in brain and that this can be observed by monitoring the 5-HIAA content of CSF.


Effective encoder-decoder neural network for segmentation of orbital tissue in computed tomography images of Graves' orbitopathy patients.
To propose a neural network (NN) that can effectively segment orbital tissue in computed tomography (CT) images of Graves' orbitopathy (GO) patients. We analyzed orbital CT scans from 701 GO patients diagnosed between 2010 and 2019 and devised an effective NN specializing in semantic orbital tissue segmentation in GO patients' CT images. After four conventional (Attention U-Net, DeepLab V3+, SegNet, and HarDNet-MSEG) and the proposed NN train the various manual orbital tissue segmentations, we calculated the Dice coefficient and Intersection over Union for comparison. CT images of the eyeball, four rectus muscles, the optic nerve, and the lacrimal gland tissues from all 701 patients were analyzed in this study. In the axial image with the largest eyeball area, the proposed NN achieved the best performance, with Dice coefficients of 98.2% for the eyeball, 94.1% for the optic nerve, 93.0% for the medial rectus muscle, and 91.1% for the lateral rectus muscle. The proposed NN also gave the best performance for the coronal image. Our qualitative analysis demonstrated that the proposed NN outputs provided more sophisticated orbital tissue segmentations for GO patients than the conventional NNs. We concluded that our proposed NN exhibited an improved CT image segmentation for GO patients over conventional NNs designed for semantic segmentation tasks.


Co-purification, co-imniunoprecipitation, and coordinate expression of acetyl-coenzyme A carboxylase activity, biotin carboxylase, and biotin carboxyl carrier protein of higher plants.
Acetyl-CoA carboxylase (ACCase; EC 126.96.36.199) is a regulatory enzyme of fatty acid synthesis, and in some higher-plant plastids is a multi-subunit complex consisting of biotin carboxylase (BC), biotin-carboxyl carrier protein (BCCP), and carboxyl transferase (CT). We recently described a Nicotiana tabacum L. (tobacco) cDNA with a deduced amino acid sequence similar to that of prokaryotic BC. We here provide further biochemical and immunological evidence that this higher-plant polypeptide is an authentic BC component of ACCase. The BC protein co-purified with ACCase activity and with BCCP during gel permeation chromatography of Pisum sativum L. (pea) chloroplast proteins. Antibodies to the Ricinus communis L. (castor) BC co-precipitated ACCase activity and BCCP. During castor seed development, ACCase activity and the levels of BC and BCCP increased and subsequently decreased in parallel, indicating their coordinate regulation. The BC protein comprised about 0.8% of the soluble protein in developing castor seed, and less than 0.05% of the protein in young leaf or root. Polypeptides cross-reacting with antibodies to castor BC were detected in several dicotyledons and in the monocotyledons Hemerocallis fulva L. (day lily), Iris L., and Allium cepa L. (onion), but not in the Gramineae species Hordeum vulgare L. (barley) and Panicum virgatum L. (switchgrass). The castor endosperm and pea chloroplast ACCases were not significantly inhibited by long-chain acyl-acyl carrier protein, free fatty acids or acyl carrier protein. The BC polypeptide was detected throughout Brassica napus L. (rapeseed) embryo development, in contrast to the multi-functional ACCase isoenzyme which was only detected early in development. These results firmly establish the identity of the BC polypeptide in plants and provide insight into the structure, regulation and roles of higherplant ACCases.


Imidazoline receptor mediated natriuresis: central and/or peripheral effect?
The ability of imidazoline agonists, such as moxonidine and rilmenidine, to lower blood pressure has been attributed to a central effect resulting in a decrease in peripheral sympathetic nerve activity. A similar decrease in sympathetic nerve activity to the kidney has been proposed to explain the increase in sodium excretion. The observed increase in sodium excretion following an intrarenal infusion of moxonidine or rilmenidine suggested the existence of a direct renal action. We therefore tested the hypothesis that direct renal infusions were acting at a central rather than a peripheral site. Thus, interventions which would decrease the natriuretic effects of central administered moxonidine would also block the effects of intrarenal administered moxonidine. Studies were performed in anesthetized Sprague-Dawley rats (280-320 g) which had undergone unilateral nephrectomy 7 to 10 days prior to the experiment. The interventions utilized resulted in minimal effects on blood pressure and creatinine clearance. Intracerebroventricular (icv) or intrarenal (ir) administration of moxonidine produced a significant increase in urine flow rate and sodium excretion. Intravenous (iv) prazosin was used to block the ability of the sympathetic nerves to alter sodium excretion secondary to alpha1-adrenoceptor stimulation. Prazosin prevented the natriuresis following icv moxonidine but only partially antagonized the effects of ir moxonidine. To determine if central imidazoline receptors mediated the effects of moxonidine, animals were pretreated with icv idazoxan. Following icv idazoxan, the effects of icv moxonidine were blocked, whereas the response to intrarenal moxonidine was only partially blocked. Peripheral (iv) administration of idazoxan blocked the actions of intrarenal moxonidine but left the response to icv moxonidine intact. Finally, chemical sympathectomy with reserpine did not alter the response to intrarenal moxonidine suggesting that this effect was independent of the sympathetic nervous system. In conclusion, these studies indicate the ability of central and peripheral moxonidine to increase urine flow rate through sodium excretion at two unique sites of action, one central and the other one peripheral, most conceivably within the kidney.


Health effects due to endotoxin inhalation (review).
Endotoxins are ubiquitous in the environment and represent important components of bioaerosols. High exposure occurs in rural environment and at several workplaces (e.g. waste collecting, textile industry etc.). Adverse effects on human health induced by inhalation of endotoxin are described in several studies. Up to now the endotoxin levels are mainly measured using the Limulus amoebocyte-lysate (LAL) assay. This assay is well established, but for a suitable characterization of bioaerosols more parameters are necessary. Additional information, e.g. concerning the pyrogenic activity of organic dust samples may be delivered by whole blood assay. Whereas on the one hand protection measures at workplaces are demanded to avoid lung function impairment due to endotoxin exposure, on the other hand a protective effect of exposure to microbial agents like endotoxins with regard to allergy development has been observed. On the cellular level toll-like receptor 4 (TLR4) and IL-1 receptor as well as surface molecules like CD14 have been shown to play a pivotal role in the endotoxin activation cascade. In this review we summarize the mechanism of endotoxin recognition and its manifold effects on human health.


Co-delivery of docetaxel and palmitoyl ascorbate by liposome for enhanced synergistic antitumor efficacy.
Palmitoyl ascorbate (PA) as an antioxidant has the potential for the treatment of cancer. In the present study, a nanocarrier system was developed for co-delivery of docetaxel (DOC) with palmitoyl ascorbate and the therapeutic efficacy of a combination drug regimen was investigated. For this purpose, different ratios of docetaxel and palmitoyl ascorbate were co-encapsulated in a liposome and they all showed high encapsulation efficiency. The average diameters of the liposomes ranged from 140 to 170 nm. Negative zeta potential values were observed for all systems, ranged from -40 mV to -56 mV. Studies on drug release and cellular uptake of the co-delivery system demonstrated that both drugs were effectively taken up by the cells and released slowly. Moreover, the liposome loading drugs with DOC/PA concentration ratio of 1:200 showed the highest anti-tumor activity to three different types of tumor cells. The higher in vivo therapeutic efficacy with lower systemic toxicity of the DOC-PA200-LPs was also verified by the H22 tumor bearing mice model. Our results showed that such co-loaded delivery systems could serve as a promising therapeutic approach to improve clinical outcomes against hepatic carcinoma.


Influence of fetal blood flow redistribution on fetal and childhood growth and fat distribution: the Generation R Study.
A suboptimal intrauterine environment leads to fetal blood flow redistribution and fetal growth restriction. Not much is known about childhood growth consequences. We examined the associations of fetal blood flow redistribution with birth outcomes, and repeatedly measured fetal and childhood growth and fat mass measures. Prospective cohort study. Population-based. One thousand one hundred and ninety-five pregnant women and their children. We measured umbilical and cerebral artery blood flow at a gestational age of 30.3 weeks (95% range, 28.5-32.6 weeks). A higher umbilical/cerebral (U/C) pulsatility index ratio is an indicator of preferential blood flow to the brain cerebral circulation at the expense of the lower body parts. Fetal and childhood growth were repeatedly measured from the third trimester until childhood. We measured the total body fat mass, lean fat mass and android/gynoid fat mass ratio by dual-energy X-ray absorptiometry and preperitoneal fat by ultrasound at 6 years. A higher fetal U/C ratio was associated with increased risks of preterm birth and small size for gestational age at birth [odds ratios, 1.41 (95% confidence interval, 1.08-1.85) and 1.63 (95% confidence interval, 1.21-2.19), respectively, per SDS increase in U/C ratio]. Longitudinal growth analyses showed that a higher fetal U/C ratio was associated with persistently lower head circumference, length and weight from third trimester fetal life until childhood (all P < 0.05). The fetal U/C ratio was not associated with total body and abdominal fat measures at 6 years. Our results suggest that fetal blood flow redistribution affects fetal development and has persistent consequences for childhood growth. Fetal blood flow redistribution affects fetal development and has persistent consequences for childhood growth.


A core mutation affecting the folding properties of a soluble domain of the ATPase protein CopA from Bacillus subtilis.
The two N-terminal domains of the P-type copper ATPase, CopAa and CopAb, from Bacillus subtilis differ in their folding capabilities in vitro. Whereas CopAb has the typical betaalphabetabetaalphabeta structure and is a rigid protein, CopAa is found to be largely unfolded. A sequence analysis of the two and of orthologue homologous proteins indicates that Ser46 in CopAa may destabilise the hydrophobic core, as also confirmed through a bioinformatic energy study. CopAb has a Val in the corresponding position. The S46V and S46A mutants are found to be folded, although the latter displays multiple conformations. S46VCopAa, in both apo and copper(I) loaded forms, has very similar structural and dynamic properties with respect to CopAb, besides a different length of strand beta2 and beta4. It is intriguing that the oxygen of Thr16 is found close, though at longer than bonding distance, to copper in both domains, as it also occurs in a human orthologue domain. This study contributes to understanding the behaviour of proteins that do not properly fold in vitro. A possible biological significance of the peculiar folding behaviour of this domain is discussed.


Topical application of a melanotropic peptide induces systemic follicular melanogenesis.
We determined the relative effectiveness of alpha-MSH and a highly potent melanotropin analogue, [Nle4, D-Phe] - alpha-MSH, in stimulating a shift from pheomelanogenesis to eumelanogenesis within hair bulbs of mice. The analogue proved to be at least a hundred times more effective than the native hormone when injected subcutaneously. The two melanotropins were then incorporated into an ointment base and topically applied to a shaved area of the skin on the back of a yellow strain of mice (C57BL/6JAY). Within 24-48 hours eumelanin production was visible within hair bulb melanocytes in both treated and untreated areas of animals. The presence of melanized organelles (eumelanosomes) within melanocytes was confirmed by electron microscopy. These results document the delivery of a peptide hormone through the skin and into the systemic circulation. This is the first demonstration of the delivery of a peptide hormone by percutaneous absorption and may provide a model for a similar route of delivery of other peptide hormones. The hormone analogue has also been delivered across human skin in vitro. Delivery of a melanotropin by a transdermal route may prove to be clinically useful in the treatment of some integumental hypopigmentary disorders in humans.


Hydroxylation of lithocholic acid by selected actinobacteria and filamentous fungi.
Selected actinobacteria and filamentous fungi of different taxonomy were screened for the ability to carry out regio- and stereospecific hydroxylation of lithocholic acid (LCA) at position 7β. The production of ursodeoxycholic acid (UDCA) was for the first time shown for the fungal strains of Bipolaris, Gibberella, Cunninghamella and Curvularia, as well as for isolated actinobacterial strains of Pseudonocardia, Saccharothrix, Amycolatopsis, Lentzea, Saccharopolyspora and Nocardia genera. Along with UDCA, chenodeoxycholic (CDCA), deoxycholic (DCA), cholic (CA), 7-ketodeoxycholic and 3-ketodeoxycholic acids were detected amongst the metabolites by some strains. A strain of Gibberella zeae VKM F-2600 expressed high level of 7β-hydroxylating activity towards LCA. Under optimized conditions, the yield of UDCA reached 90% at 1g/L of LCA and up to 60% at a 8-fold increased substrate loading. The accumulation of the major by-product, 3-keto UDCA, was limited by using selected biotransformation media.


In vitro inhibition of human malignant brain tumour cell line proliferation by anti-urokinase-type plasminogen activator monoclonal antibodies.
A brain tumour-associated marker, urokinase (UK), was investigated using rabbit anti-UK polyclonal and murine anti-UK monoclonal antibodies, which were prepared by immunization with low molecular weight UK (LMW-UK) and high molecular weight urokinase (HMW-UK) synthetic peptide respectively. The polyclonal antibody cross-reacted with both LMW-UK and HMW-UK, whereas the murine MAbs were specific for HMW-UK. These immunological probes were used to study urokinase in glioma extracts, tissues, sera and cell lines that had been prepared from primary cultures of freshly dissected gliomas. Radioimmunoassays showed that glioma extracts had much higher level (5- to 44-fold) of UK than normal human brain extracts. This result was confirmed by immunoblotting of electrophoresis gels of glioma and human brain extracts. Immunohistochemical study using anti-UK MAb demonstrated much higher levels of UK in glioma tissue than normal brain tissue. Immunohistochemical study using anti-UK MAbs localized UK on the cell surface of glioma cells. Anti-UK MAbs inhibited the proliferation of AA cell lines and GB cell lines (50% to > 90%) and exerted minor effects (< or = 20%) on normal human liver, intestine and lymphocyte cell lines. Taken together, these results suggest that anti-UK MAbs may have therapeutic potential for human gliomas and cancer metastasis.


Multifunctional Monoamine Oxidases and Cholinesterases Inhibitory Effects, as well as UPLC-DAD-MS Chemical Profile of Alkaloid Fractions Obtained from Species of the Palicoureeae Tribe.
In the present study, the effects were evaluated of alkaloid fractions (AFs) from Psychotria species and correlated genera, Palicourea and Rudgea, on monoamine oxidases (MAOs) and cholinesterases (ChEs). By HPLC-DAD and UPLC-DAD-MS analyses, indole alkaloids (IA) were detected in all AFs. For the Psychotria and Palicourea species, these IA corresponded to tetrahydro-p-carboline alkaloids (THPCA). On the other hand, pyrrolidinoindoline core compounds were observed for Rudgea species. Regarding their pharmacological activities, none of the AFs was able to inhibit AChE. However, the BChE activity was impaired by the Psychotria and Palicourea AFs. In addition, MAO-A was inhibited by both AFs, but only Psychotria nemorosa AF was able to inhibit significantly MAO-B. Rudgea AFs demonstrated a poor inhibitory profile on MAO-A. Taken together, our results highlighted the Psychotria and Palicourea genera as important sources of scaffolds for the development of MAO-A and BChE inhibitors aiming at the treatment of neurodegenerative and neuropsychiatric diseases.


Synchronous CIN 3 and cervical lymphoma: a case report and review of the literature.
Primary cervical lymphoma is rare, with less than 60 cases reported in the English literature, including a few cases of synchronous cervical intraepithelial neoplasia (CIN) with cervical lymphoma. We describe a 42-year-old woman with CIN 3 and malignant lymphoma of the cervix. The Papanicolaou cytology only revealed dysplastic cells of CIN 3, but not the malignant lymphoid cells. The histology of the cervix revealed diffuse infiltration of monotonous medium-sized lymphoid cells with dispersing tingible bodies, and a Burkitt's lymphoma in combination with a small CIN 3 lesion was diagnosed. Neither human papillomavirus nor Epstein-Barr virus was identified by polymerase chain reaction or in situ hybridization. The patient developed leukemic spreading and died soon after surgery.


Three years of neonatal morbidity and mortality at the national hospital in Dili, East Timor.
The aim of this study was to undertake a retrospective review of admissions and discharges to the neonatal unit at the National Hospital Guido Valadares, Dili, in order to gain insight into the epidemiology of hospitalised neonates in East Timor, as the information cannot be obtained from the hospital health management information system. Data were sourced from unit registers for 3 years, 2008-2010 inclusive. Demographic characteristics and diagnoses were related to the risk of dying using stepwise multivariate logistic regression and adjusting for potential confounders of age, sex and weight. Two thousand eighty-eight babies were admitted to the unit over the study period. Over a quarter of babies weighed <2.5 kg on admission. Almost half were admitted from emergency or outpatient departments and only 27% were admitted within their first week of life. The most common reasons for admission were sepsis and respiratory disease (38 and 22%, respectively). Overall mortality was 11.4%, mainly attributed to prematurity (28%), infection (26%) and asphyxia (24%). Home birth, male gender, very low weight, young age and a short duration of hospitalisation were independently associated with an increased risk of death. Half of all babies weighing <1.5 kg died. Two-thirds of deaths occurred within 2 days of admission. The study provides, for the first time, an insight into the admissions and outcomes of the largest neonatal unit in East Timor. It is a baseline from which improvements to the quality of clinical care and data collection can be made.


Misdiagnosed localized arteriovenous malformation.
We report a case of an 11-year-old girl with a pulsatile mass in the thigh diagnosed by angiography as localized arteriovenous malformation. Recent increase in tenderness and size of the lesion brought the patient to hospital and excision was performed at the parents' request. Subsequent histologic examination revealed that this lesion was an alveolar soft-part sarcoma in disguise. The angiographic and pathologic results are presented. Other investigators should be aware of this uncommon and highly vascular sarcoma, which might be misdiagnosed as a localized arteriovenous malformation and thus treated only conservatively.


Therapeutic potential of uracil and its derivatives in countering pathogenic and physiological disorders.
Uracil is one of the most notable pharmacophores in medicinal chemistry as the pyrimidine nucleobase forms an integral part of many commercial drugs. Though the name uracil is usually associated with cancer drugs, there are many uracil-based compounds which can treat different diseases when they are employed. So far, there has been no in-depth review concerning uracil drugs in the market, or in the different stages of clinical trials, including those approved or discontinued. The current work focuses on the importance of uracil and its derivatives in treating different diseases. The use of uracil compounds in treating viral infections, cancer, diabetic, thyroid and autosomal recessive disorders are discussed in the review. The mechanism of action of each uracil drug with emphasis on their structure and properties are discussed in detail. The targeted action of these drugs on sites or on the different stages of a disorder/pathogenic life cycle are also discussed. This review encompasses uracil drugs approved as well as those in development from the 1950's onwards. The utility of uracil in drug discovery and its association with a wide range of diseases is brought forth within this review to demonstrate its potential to a wider audience.


Advantages of CEMiTool for gene co-expression analysis of RNA-seq data.
Gene co-expression analysis is widely applied to transcriptomics data to associate clusters of genes with biological functions or identify therapeutic targets in diseases. Recently, the emergence of high-throughput technologies for gene expression analyses allows researchers to establish connections through gene co-expression analysis to identify clinical disease markers. However, gene co-expression analysis is complex and may be a daunting task. Here, we evaluate three co-expression analysis packages (WGCNA, CEMiTool, and coseq) using published RNA-seq datasets derived from ischemic cardiomyopathy and chronic obstructive pulmonary disease. Results show that the packages produced consensus co-expression clusters using default parameters. CEMiTool package outperformed the other two packages and required less computational resource and bioinformatics experience. This evaluation provides a basis on which data analysts can select bioinformatics tools for gene co-expression analysis.


Phospholipid and fatty acid composition of tetrodotoxin receptor-rich membrane fragments from Electrophorus electricus.
To study the interaction of voltage-sensitive Na+-channels with membrane lipids, the phospholipid and fatty acid composition of highly purified membrane fragments from the remarkably differentiated plasma membrane of Electrophorus electricus has been analyzed. After density gradient fractionation and carrier free electrophoresis, fractions with up to 30 pmol tetrodotoxin binding/mg protein can be obtained, which may correspond to a 50% pure preparation of the extrasynaptic part of the excitable face. Phospholipid classes and cholesterol are separated by one-dimensional thin-layer chromatography in acidic and alkaline solvent systems. The following mean molar contents are found: 40% phosphatidylcholine, 23% phosphatidylserine, 30% phosphatidylethanolamine and 7% sphingomyelin. In a series of 11 animals, significant deviations from these mean values have been observed. The fatty acid composition of the phospholipids has been determined by gas chromatography. Phosphatidylcholine contains more than 50% 16:0, and about 20% unsaturated fatty acids in the C-18 group. Compared to other plasma membrane fractions, this phospholipid is the least differentiated. By contrast, phosphatidylethanolamine and phosphatidylserine show many characteristics in different membrane fractions, especially in their unsaturated components representing more than 50%. 22:6, as the major constituent in these fractions, accounts for a quarter to a third of all fatty acids in these fractions. 18:0 is the main saturated component in these two phospholipids with abundances of typically a quarter or less of all fatty acids. Knowledge of the lipid composition of these excitable membranes may help to conserve binding and structural properties when analyzing lipid-sensitive Na+-channels in vitro. It is also useful as a guideline for systematic reconstitution studies.


Antipsychotic-like profile of combined treatment with raclopride and 8-OH-DPAT in the rat: enhancement of antipsychotic-like effects without catalepsy.
The administration of the 5-HT1A agonist 8-OH-DPAT, 0.1 mg kg-1 sc-20 min, produced a moderate suppression of conditioned avoidance behavior (60% of controls) in the rat. This effect, however, was not seen after administration of higher doses, 0.4 and 1.6 mg kg-1 sc. The number of intertrial crosses were not affected by the lower dose but significantly increased by administration of the two higher doses of 8-OH-DPAT. The dopamine D2 receptor blocking agent raclopride, 0.05 mg kg-1, by itself did not suppress the avoidance behavior, but in combination with 8-OH-DPAT produced suppression of avoidance behavior (30% of controls) as well as intertrial crosses. Open field locomotor activity was suppressed by raclopride, 0.1 mg kg-1 sc, or by 8-OH-DPAT, 0.1 mg kg-1 sc. The combined treatment produced a further suppression of locomotor activity and a marked increase in "immobility" (stationary movements). Treadmill locomotion, however, was not affected by either compound by itself, whereas the combined treatment impaired treadmill performance. Suppression of treadmill performance by a higher dose of raclopride, 0.4 mg kg-1 sc, was not altered by the additional treatment with 8-OH-DPAT, 0.1 mg kg-1. In contrast to the additive effects of 8-OH-DPAT and raclopride on conditioned avoidance behavior, open field locomotion and treadmill performance, the catalepsy produced by raclopride, 16 mg kg-1 was completely antagonised by treatment with 8-OH-DPAT 0.1 mg kg-1. Taken together, the present findings demonstrate strong interactions between a 5-HT agonist and a DA D2 antagonist on some critical tests for antipsychotic-like actions and extrapyramidal motor effects in rats, and suggest new possibilities in the search for new antipsychotic drugs with higher clinical efficacy and less extrapyramidal side effects.


Patient satisfaction and ward atmosphere during a crisis in an open psychiatric ward.
There is great need for useful indicators of quality of care for inpatient psychiatric treatment. The study examines whether patient satisfaction and ward atmosphere can be used as a means of evaluating an assumed decline in quality of care during a crisis in an open psychiatric ward. Twice a year for 2 years ten patients filled out a standardized patient satisfaction questionnaire developed by the Swedish Institute for Health Services Development (SPRI) on their day of departure. At the same time all the patients in the ward evaluated the ward atmosphere using the Ward Atmosphere Scale (WAS). In the third study period the ward went through a crisis where quality of care was expected to be lower. The SPRI questionnaire confirmed the decline in patients' perceived quality of care on six of 41 questions, WAS was markedly lower on five of ten subscales. Both WAS and the SPRI questionnaire gave useful information on how the ward was affected by the crisis. WAS seemed to be a more sensitive way of measuring quality of care, while the SPRI questionnaire indicated more clearly the practical consequences.


The enhancement of cyclosporin A-induced nephrotoxicity by gentamicin.
Normal rats receiving treatment with either cyclosporin A or gentamicin show dose-dependent nephrotoxicity, the changes with cyclosporin A centering on the straight segment of the proximal tubule, while gentamicin affects the convoluted segment. When the two drugs are given together, nephrotoxicity is enhanced with significant reductions in renal function even at therapeutic dosage. The study emphasises the caution needed when potentially nephrotoxic agents are administered in combination.


Amantadine treatment and delayed onset of levodopa-induced dyskinesia in patients with early Parkinson's disease.
Levodopa-induced dyskinesia (LID) is a common motor complication in patients with Parkinson's disease (PD). Although amantadine is indicated for LID treatment, it is uncertain whether early treatment with amantadine reduces the risk of LID in patients with PD. We aimed to evaluate the association between amantadine treatment and LID onset in patients with early-stage PD. This was a hospital-based retrospective cohort study that used electronic medical records from January 1, 2009 to October 31, 2016. The effect of amantadine on LID onset was compared with those of anticholinergics and monoamine oxidase type B inhibitors in patients with PD. Propensity-score weighting and landmark analysis were used to reduce potential confounding. The time to LID onset was analyzed using Cox models. Sensitivity analyses were performed to determine the robustness of the results. The analyses included 807, 661, and 518 patients at 6-, 12-, and 18-month landmark points, respectively. Amantadine use was associated with delayed LID onset in the 6- and 12-month landmark analyses, with adjusted hazard ratios of 0.65 (95% confidence interval [CI] = 0.49-0.86) and 0.64 (95% CI = 0.47-0.88), respectively. Sensitivity analysis findings were comparable to those of the main analysis. Early treatment with amantadine may delay LID onset more than treatment with other symptomatic agents. Further studies are needed to elucidate the mechanism of amantadine in LID onset delay and to validate our findings.


Characteristics of retained foreign bodies and near-miss events in the operating room: a ten-year experience at one institution.
Retained foreign bodies (RFBs) are a major complication of surgical procedures. However, the efficacy of preventive measures is not well defined. This study investigates the characteristics of potential (near miss) and actual RFBs, and the contributions of routine practice for the prevention of RFB events. We conducted a retrospective review of incident reports regarding near-miss and RFB events in patients who underwent surgery under general anesthesia in our institution between October 2008 and November 2018. Among 49,831 operations under general anesthesia, there were 106 (2.13/1000) near-miss events and 24 (0.48/1000) RFB events. Counting surgical materials and intraoperative X-rays detected the remaining items before completion of surgery in 59 (56%) and 15 (14%) cases, respectively. The operator or staff noticed the surgical materials in the remaining 32 (30%) near-miss events. RFBs included 4 sponges (17%), 4 instruments (17%), 4 needles (17%), and 12 miscellaneous items (50%). Of these, 12 (50%) RFBs were discovered on postoperative X-rays and 16 (67%) patients required operative removal. Four incidents (17%) with RFBs were attributable to ignoring count discrepancies during surgery. The actual incidence of RFB events is higher than previously reported. A standardized counting protocol, communication among staff, and intra- and postoperative X-rays may contribute to the prevention and detection of RFBs.


[IgG responses to candidate malaria vaccine antigens in the urban area of Dakar (Senegal): evolution according to age and parasitemia in patients with mild symptoms].
Malaria remains a major problem in African countries despite substantial decreases in morbidity and mortality due to sustained control programs. Studies for the evaluation of qualitative or quantitative Ab responses to key targets of anti-plasmodium immunity were mostly done in rural endemic setting compared to urban area. In a cohort of 200 patients with mild malaria and living in Dakar, we analyze total and subclasses IgG responses to a panel of P. falciparum blood stage antigens: MSP1p19, MSP3, EB200, GST-5 and R23. A mean age of 15 yrs (4 to 56 yrs) and parasitemia between 0.1 to 17% were found. Levels of IgG anti-MSP3 were higher in patients with low parasitemia (≤1%) and appear negatively correlated to parasite densities (Rho =. 0.54; p= 0.021). This correlation is more significant in children (≤ 15 yrs). In addition, an increase of IgG responses against MSP1p19 is highly observed in adults having a parasitemia less than 1%. In those patients, we find that IgG1 subclasses were predominant (p <0.01). Our study shows an association between Ab responses and parasitemia. This association is dependant to IgG anti-MSP3 in children and IgG anti-MSP1p19 in adults living in urban area.


Aberrant thymus in infants: sonographic evaluation.
We report five children with aberrant thymus, four of whom presented with an upper suprasternal mass and one with a lateral neck mass. Awareness of this entity, together with knowledge of its variable presentation, is essential for unnecessary surgery to be avoided. US should be the initial diagnostic approach and occasionally leads to the correct diagnosis.


Caries and fluorosis prevalence in communities with different concentrations of fluoride in the water.
The need to defluoridate and fluoridate the water supplies in areas with drinking water naturally containing above-optimal (>/=2.5 mg/l) and suboptimal (</=0.3 mg/l) fluoride concentration and caries and fluorosis prevalence of 12-year-old schoolchildren were assessed in Italy. In the low-fluoride area, 48.4% children were caries-free (DMFT = 0) and the DMFT and DMFS were 1.5 and 2.6; in the high-fluoride area, 46.8% had a DMFT = 0 and the values of the indices were 1.4 and 1.6, respectively. Multiple logistic regression analysis showed a significant association in the caries-free status according to parents' employment status (OR = 1.2, 95% CI = 1.1-1.3) and children's sweets consumption, since children who consumed sweets at least once a day had an adjusted odds ratio of 1.8 (95% CI = 1.4-2.3) compared to those with a lower consumption. Multiple linear regression analysis showed that DMFT and DMFS were significantly higher in children with a lower socioeconomic status and in those who consumed sweets at least once a day, with the DMFS significantly associated also with the area of residence. DT and FT scores were higher in the high- and low-fluoride areas, respectively. No evidence of fluorosis was reported in 94.5 and 55.3% of children in the low- and high-fluoride areas, respectively. The Community Fluorosis Index (CFI) for all permanent teeth was significantly higher in the high-fluoride area, 0.8, than the value, 0.1, found in the low-fluoride community. Our results substantiate the difficulties in defining universal guidelines for the fluoridation or defluoridation of drinking water and the need for an epidemiological approach to the decision as to fluoridate and defluoridate the water supply.


Lumbar plexus terminal branch block, a safe alternative for transfemoral aortic valve implantation. Case report.
Transcatheter aortic valve implantation is an alternative treatment for patients with severe aortic stenosis, it is conventionally performed under general anaesthesia or local anaesthesia plus sedation. We present the first case of trans-femoral, trans-catheter aortic valve implantation, performed in our hospital in a patient with severe aortic stenosis, who was a high surgical risk. Anaesthesia consisted of a combination of bilateral selective blockade of the iliohypogastric, ilioinguinal and genitofemoral nerves with the patient awake without sedation, using an ultrasound-guided approach. Transcatheter aortic valve implantation was successful and passed without incident, the patient remained immobile, calm, did not report pain, and sedation or hypnotics were not necessary during dilation of the vascular accesses with the introducer. Standard monitoring demonstrated haemodynamic stability, without cardiovascular repercussions and did not necessitate vasopressor or inotropic drug support. After the intervention, the patient was sent to the Cardiac Intensive Care Unit, where he remained asymptomatic and stable. Subsequently, the patient was admitted to the cardiology ward from where he was discharged without complications.


Drug-induced osteoporosis: beyond glucocorticoids.
To prevent osteoporotic fractures, it is important to identify patients at high risk of fracture by assessing risk factors such as underlying diseases and drugs. Prior and current use of oral corticosteroids is strongly associated with increased fracture risk, but other medications could be involved and should be assessed as part of the optimal care of patients at risk of osteoporosis. Strong evidence points to gonadotropin-releasing hormone agonists, aromatase inhibitors, and antiepileptic drugs. Further studies are necessary to confirm the deleterious effect of other treatments on bone.


Sugarbeet mitochondria contain an open reading frame showing extensive sequence homology to the subunit 2 gene of the NADH: ubiquinone reductase complex.
Sequence analysis of a transcribed region of mitochondrial DNA (mtDNA) from male fertile sugarbeet (Beta vulgaris L.) revealed an open reading frame showing extensive sequence homology to the subunit 2 gene of the NADH: ubiquinone reductase complex (nad2). Sugarbeet nad2 in common with its proposed counterpart in animal mitochondria has no intron and thereby differs from the corresponding chloroplast gene. Northern RNA analysis of sugarbeet nad2 suggested that transcription of this locus gives rise to at least three transcripts. No differences in transcript profile were detected between male fertile and cytoplasmic male sterile sugarbeet. This constitutes the first report of a mitochondrial nad2 gene in higher plants.


In silico and in vitro safety assessment of a fungal biomass from Rhizomucor pusillus for use as a novel food ingredient.
To address the growing world population and reduce the impact of environmental changes on the global food supply, ingredients are being produced using microorganisms to yield sustainable and innovative products. Food ingredients manufactured using modern biotechnology must be produced by non-toxigenic and nonpathogenic production organisms that do not harbor antimicrobial resistance (AMR). Several fungal species represent attractive targets as sources of alternative food products. One such product is a fungal biomass obtained from the fermentation of Rhizomucor pusillus strain CBS 143028. The whole genome sequence of this strain was annotated and subjected to sequence homology searches and in silico phenotype prediction tools to identify genetic elements encoding for protein toxins active via oral consumption, virulence factors associated with pathogenicity, and determinants of AMR. The in silico investigation revealed no genetic elements sharing significant sequence homology with putative virulence factors, protein toxins, or AMR determinants, including the absence of mucoricin, an essential toxin in the pathogenesis of mucormycosis. These in silico findings were corroborated in vitro based on the absence of clinically relevant mycotoxin or antibacterial secondary metabolites. Consequently, it is unlikely that R. pusillis strain CBS 143028 would pose a safety concern for use in food for human consumption.


Effects of Marantodes pumilum (Kacip Fatimah) on vaginal pH and expression of vacoular ATPase and carbonic anhydrase in the vagina of sex-steroid deficient female rats.
In sex-steroid deficiency, increased in the pH of vaginal fluid is due to low estrogen levels. Consumption of Marantodes pumilum leaves helps to ameliorate increased in vaginal fluid pH in sex-steroid deficient condition. To investigate changes in vaginal fluid pH and expression of proteins that participate in pH changes i.e vacoular (V)-ATPases and carbonic anhydrases (CA) in the vagina following M. pumilum leaves consumption. Ovariectomized adult female rats were treated orally with M. pumilum leaves extract (MPE) at 100, 250 and 500 mg/kg.b.w and estradiol at 0.2 µg/kg/b.w for 28 days. At the end of the treatment, vaginal fluid pH was measured in anesthetised rats by using micropH probe. Following sacrificed, levels of V-ATPase and CA proteins and mRNAs in the vagina were identified by Western blotting and real-time PCR, respectively. Protein distribution was visualized by immunohistochemistry. Administration of MPE causes the pH of vaginal fluid to decrease and expression and distribution of vaginal V-ATPase A & B and CA II, III, IX, XII and XIII to increase. The decrease in vaginal fluid pH following MPE treatment suggested that this herb has potential to be used to ameliorate vaginal fluid pH changes in sex-steroid deficient condition.


Cystic paraganglioma of the anterior mediastinum.
Paraganglioma of the mediastinum is an extremely uncommon neoplasm, and the concurrent finding of cystic changes has yet to be reported in the literature. We report a case of a 43-year-old man presenting with chest pain and left arm paresthesias. Imaging studies revealed a well-defined cystic lesion located in the anterior mediastinum, with the patient undergoing subsequent surgical resection of the mass. Grossly, the heterogenous mass measured 6.0 × 3.6 × 1.7 cm, with prominent cystic spaces and scattered solid areas. Microscopic examination revealed pseudocystic spaces with proliferation of large epithelioid cells with abundant eosinophilic cytoplasm, moderate pleomorphic nuclei, and prominent nucleoli, in a nested pattern of growth along the cysts walls. Immunohistochemical stains showed strong positivity for chromogranin, synaptophysin, and CD56 in the epithelioid cells, with negative staining for cytokeratin AE1/AE3, confirming the diagnosis of paraganglioma. S-100 was positive in the intervening sustentacular cells throughout the tumor. Cystic paraganglioma of the anterior mediastinum is a rare neoplasm, which , to our knowledge, has not yet been reported in the English literature, and should be added to the differential diagnosis of cystic lesions of the mediastinum.


Loss of rapid phonological recoding in reading Hanja, the logographic script of Korean.
The role that recent experience has in the processing of Korean Hanja characters was investigated in two masked priming experiments. Two groups of Korean native speakers that differed in their recent exposure to Hanja were asked to name single Hanja characters (targets) that were immediately preceded by masked presentations of the same characters (repetition priming), Hanja characters that were homophones of the target (homophone priming), or unrelated characters (baseline). The results showed that the group that had been less recently exposed made more errors and were slower than the group that had been the more recently exposed. Furthermore, there was no character homophone priming for the less recently exposed group, although they did show a robust repetition priming effect. On the other hand, the more recently exposed group showed both strong character repetition and homophone priming effects. We suggest that regular exposure to characters supports their rapid and automatic processing. It is argued that the different patterns of priming for the two groups were due to how rapidly the orthographic and phonological information of the prime could be resolved.


pH-sensitive nanocarrier based on gold/silver core-shell nanoparticles decorated multi-walled carbon manotubes for tracing drug release in living cells.
We fabricate a multifunctional nanocarrier based on multi-walled carbon nanotubes (MWCNTs) decorated with gold/silver core-shell nanoparticles (Au@Ag NPs) and fluorescein isothiocyanate (FITC) for tracking the intracellular drug release process. In the demonstrated nanocarrier, the Au@Ag NPs adsorbed on the surface of MWCNTs were labeled with the pH-dependent SERS reporter 4-Mercaptobenzoic acid (4MBA) for SERS based pH sensing. FITC was conjugated on MWCNTs to provide fluorescence signal for tracing the MWCNTs. Fluorescent doxorubicin (DOX) was used as the model drug which can be loaded onto MWCNTs via π-π stacking and released from the MWCNTs under acidic condition. By detecting the SERS spectrum of 4MBA, the pH value around the nanocarrier could be monitored. Besides, by tracing the fluorescence of FITC and DOX, we can also investigate the drug release process in cells. Experimental results show that the proposed nanocarrier retained a well pH-sensitive performance in living cells, and the DOX detached from MWCNTs inside the lysosomes and entered into the cytoplasm with the MWCNTs being left in lysosomes. To further investigate the drug release dynamics, 2-D color-gradient pH mapping were plotted, which were calculated from the SERS spectra of 4MBA. The detailed release process and carrier distribution have been recorded as environmental pH changes during cell endocytosis. Furthermore, we also confirmed that the proposed nanocarrier has a good biocompatibility. It indicates that the designed nanocarrier have a great potential in intraceable drug delivery, cancer cells imaging and pH monitoring.


Drowning Injuries: Analysis of a Decennial Air Medical Rescue Center Experience.
Drowning is one of the leading injury death causes in younger children. Common intensive care measures seem not to improve neurologic outcome, and early prognostic options appear partially unreliable. Therefore, we evaluated a cohort of drowning patients cotreated by a helicopter emergency medical service regarding typical incident constellations, early and subsequent prognostic options, and relevant interventions. All patients prehospitally cotreated by helicopter emergency medical service "Christoph 4" in primary missions because of drowning incidents during a 10-year period were evaluated. Patient, prehospital, and clinical data were recorded retrospectively; correlations and prognostic values were evaluated with appropriate statistical tests. Fifty-one patients were included. Various examination results (several vital, neurologic, and laboratory parameters) and sufficient prehospital first aid measures were significantly correlated with the final outcome (P < .05, respectively). Aspartate aminotransferase and alanine aminotransferase values precisely discriminated between the final outcome groups (P = .001 and area under the receiver operating characteristic curve = 1.0 in both cases). Aspartate aminotransferase and alanine aminotransferase values were the most useful predictors of outcome in our study. Sufficient prehospital first aid measures were correlated with improved outcome. Regular first aider training is recommended.


Aqua-cyanido{6,6'-dimeth-oxy-2,2'-[1,2-phenyl-enebis(nitrilo-methanylyl-idene)]diphenolato}cobalt(III) acetonitrile hemisolvate.
In the title complex, [Co(C(22)H(18)N(2)O(4))(CN)(H(2)O)]·0.5CH(3)CN, the Co(III) cation is N,N',O,O'-chelated by a 6,6'-dimeth-oxy-2,2'-[1,2-phenyl-enebis(nitrilo-methanylyl-idene)]diphenolate dianion, and is further coordinated by a cyanide anion and a water mol-ecule in the axial sites, completing a distorted octa-hedral coordination geometry. In the crystal, pairs of bifurcated O-H⋯(O,O) hydrogen bonds link adjacent mol-ecules, forming centrosymmetric dimers. The acetonitrile solvent mol-ecule shows 0.5 occupancy.


Indian Academy of Pediatrics Revised (2021) Guidelines on Prevention and Treatment of Vitamin D Deficiency and Rickets.
The emerging literature on prevalence of vitamin D deficiency in India, prevention and treatment strategies of rickets, and extra-skeletal benefits of vitamin D suggest the need for revising the existing guidelines for prevention and treatment of vitamin D deficiency in India. To review the emerging literature on vitamin D prevalence and need for universal vitamin D supplementation. To suggest optimum vitamin D therapy for treatment of asymptomatic and symptomatic vitamin D deficiency, and rickets. To evaluate the extra-skeletal health benefits of vitamin D in children. A National consultative committee was formed that comprised of clinicians, epidemiologists, endocrinologists, and nutritionists. The Committee conducted deliberations on different aspects of vitamin D deficiency and rickets through ten online meetings between March and September, 2021. A draft guideline was formulated, which was reviewed and approved by all Committee members. The group reiterates the serum 25-hydroxy vitamin D cutoffs proposed for vitamin D deficiency, insufficiency, and sufficiency as <12 ng/mL, 12-20 ng/mL and >20 ng/mL, respectively. Vitamin D toxicity is defined as serum 25OHD >100 ng/mL with hypercalcemia and/or hypercalciuria. Vitamin D supplementation in doses of 400 IU/day is recommended during infancy; however, the estimated average requirement in older children and adolescents (400-600 IU/day, respectively) should be met from diet and natural sources like sunlight. Rickets and vitamin D deficiency should be treated with oral cholecalciferol, preferably in a daily dosing schedule (2000 IU below 1 year of age and 3000 IU in older children) for 12 weeks. If compliance to daily dosing cannot be ensured, intermittent regimens may be prescribed for children above 6 months of age. Universal vitamin D supplementation is not recommended in childhood pneumonia, diarrhea, tuberculosis, HIV and non-infectious conditions like asthma, atopic dermatitis, and developmental disorders. Serum 25-hydroxy vitamin D level of >20 ng/mL should be maintained in children with conditions at high-risk for vitamin deficiency, like nephrotic syndrome, chronic liver disease, chronic renal failure, and intake of anticonvulsants or glucocorticoids.


Corneoscleral ulceration and corneal perforation after pterygium excision and topical mitomycin C therapy.
A 78-year-old woman complained of ocular pain in the right eye. Thirty-three months before, the patient had a pterygium excised and mitomycin C, 0.04%, was instilled 3 times a day postoperatively for 10 days in the right eye. The same surgery was performed and the same drug was used for a course of 5 days in the left eye. Corneoscleral ulceration and corneal performation occurred in the right eye. The lesions were successfully treated with a lamellar patch graft with reserved corneosclera. We believe that it may not be safe to instill 0.04% mitomycin C 3 times a day postoperatively for 10 days.


Pyrosequencing reveals highly diverse microbial communities in microbial electrolysis cells involved in enhanced H2 production from waste activated sludge.
Renewable H(2) production from a plentiful biomass, waste activated sludge (WAS), can be achieved by fermentation, but the yields are low. The use of a microbial electrolysis cell (MEC) can increase the H(2) production yields to several times that of fermentation. We have proved that the enhancement of H(2) production was due to the ability of MECs to use a wider range of organic matter in WAS than in fermentation. To support this result strongly, we here investigated the microbial community structures of WAS and anode biofilms in WAS-fed MECs. A pyrosequencing analysis based on the bacterial 16S rRNA gene showed that dominant populations in MECs were more diverse than those in WAS (inoculum and substrate) after enrichment, and there was a clear distinction between MECs and WAS in microbial community structure. Diverse acid-producing bacteria and exoelectrogens (predominance of Geobacter) were detected in MECs but they were only rarely found in WAS. It has been reported that these acid-producing bacteria can ferment various sugars and amines with acetate, propionate, and butyrate as their major by-products. This was consistent with our chemical analyses. Detected exoelectrogens are known to use these organic acids (mainly acetate) and certain sugars to directly produce current for H(2) generation at the cathodes in the MECs. Using quantitative real-time PCR, we demonstrated that a consistent feed of alkaline-pretreated WAS containing large amounts of acetate led to a predominance of acetoclastic methanogens, while hydrogenotrophic methanogens were abundant in MECs fed both raw and alkaline-pretreated WAS. Syntrophic interactions between phylogenetically diverse microbial populations in anodophilic biofilms were found to drive the efficient cascade utilization of organic matter in WAS.


An unusual effect of N-octylamine on the cytochrome b5 spectrum of insect and mammalian microsomes.
The addition of n-octylamine to microsomes prepared from the midgut of tobacco hornworn (Manduca sexta) larvae causes an unusual spectral interaction. The initial optical difference spectrum appears to be the sum of reduced cytochrome b5 and a type II difference spectrum of cytochrome P-450. This initial spectrum is unstable and diminishes in size, with a concurrent shift in peak (424 to 428 nm) and trough (409 and 392 to approx. 400 nm) positions, to yield a stable spectrum identical to the type II spectrum of cytochrome P-450. Thus, in addition to its interaction with cytochrome P-450, n-octylamine causes a reduction of cytochrome b5 which subsequently becomes reoxidized. The causal factor for this unusual spectral interaction occurs in the cytoplasm and appears to be protein-bound. It was also present in similar preparations from the tobacco budworm (Heliothis virescens) but not in those from rat or mouse liver or abdomens from insecticide-resistant or susceptible houseflies (Musca domestica). Microsomes from rat and mouse liver, but not those from housefly abdomens, exhibit similar unusual spectral interactions with n-octylamine when supplemented with the soluble factor from the hornworm.


The mechanisms associated with the development of hypertension after exposure to lead, mercury species or their mixtures differs with the metal and the mixture ratio.
Hypertension is considered to be the most important risk factor for the development of cardiovascular diseases. Beside life-style risk factors, exposure to lead and mercury species are increasingly discussed as potential risk factors. Although there are a few previous studies, the underlying mechanism by which exposure to lead and mercury disturb blood pressure regulation is not currently understood. Potential mechanisms are oxidative stress production, kidney damage and activation of the renin-angiotensin system (RAS), all of which can interact to cause dysregulation of blood pressure. Male rats (Wistar) were exposed to lead, inorganic mercury, methylmercury or two mixtures of all three metals for four weeks through the drinking water. The two mixture ratios were based on ratios of known reference values or environmental exposure from the literature. To investigate the potential mechanism of actions, blood pressure was measured after four weeks and compared to plasma nitrotyrosine or reduced/oxidized glutathione levels in liver as markers for oxidative stress. Plasma renin and angiotensin II levels were used as markers for RAS activation. Finally, kidney function and injury were assessed via urinary and plasma creatinine levels, creatinine clearance and urinary kidney-injury molecule (KIM-1). While exposure to lead by itself increased oxidative stress and kidney damage along with blood pressure, inorganic mercury did not affect blood pressure or any end-point examined. Conversely, methylmercury instead increased RAS activation along with blood pressure. Surprisingly, when administered as mixtures, lead no longer increased oxidative stress or altered kidney function. Moreover, the mixture based on an environmental ratio no longer had an effect on blood pressure, while the reference value ratio still retained an increase in blood pressure. Based on our results, the prominent mechanism of action associated with the development of hypertension seems to be oxidative stress and kidney damage for lead, while increased RAS activation links methylmercury to hypertension, but these mechanisms along with hypertension disappear when metals are present in some mixtures.


Implementing peer review at an emergency medicine blog: bridging the gap between educators and clinical experts.
Emergency physicians are leaders in the ''free open-access meducation'' (FOAM) movement. The mandate of FOAM is to create open-access education and knowledge translation resources for trainees and practicing physicians (e.g., blogs, podcasts, and vodcasts). Critics of FOAM have suggested that because such resources can be easily published online without quality control mechanisms, unreviewed FOAM resources may be erroneous or biased. We present a new initiative to incorporate open, expert, peer review into an established academic medical blog. Experts provided either pre- or postpublication reviews that were visible to blog readers. This article outlines the details of this initiative and discusses the potentially transformative impact of this educational innovation.


Lower Extremity Osteoarthritis: Management and Challenges.
Osteoarthritis is the most common form of arthritis, a significant contributor to disability and the major cause for joint replacements, but treatment for the disease is still limited. Although no disease-modifying treatments exist, there are evidence-based algorithms and guidelines for the symptomatic treatment of the disease.


Structural and functional diversity of copper-metallothioneins from the American lobster Homarus americanus.
The role of copper metallothionein (CuMT) in copper metabolism and metalloenzyme activation is poorly understood. We have chosen marine crustaceans, in which a direct correlation exists between levels of Cu(I)MT and Cu(I)-hemocyanin during the molt cycle (Engel and Brouwer, Biol. Bull. 173, 239-251, 1987) as unique model systems to study the involvement of MTs in metalloprotein activation and degradation. We have isolated three low-molecular weight, cysteine-rich copper proteins from the American lobster Homarus americanus, which we designate as CuMT-1, CuMT-2, and CuMT-3, respectively. As a first attempt to fully characterize these proteins, we have determined the sequence of the first 56 amino acids of CuMT-1. The results show this protein to belong to the class I MTs, i.e., related in primary structure to equine renal MT. CuMT-1 cannot transfer its copper to copper-depleted apohemocyanin. CuMT-2 belongs to the same class of MTs as CuMT-1, but CuMT-3 does not. The latter can reactivate lobster hemocyanin containing reduced amounts of Cu(I). Spectroscopic studies show that Cu(I) transfer from CuMT-3 to apohemocyanin initially results in the formation of distorted binuclear-copper sites, which subsequently slowly return to their native stereochemical configuration. Finally, we present evidence that shows that the class I MTs in marine crustacea are involved in the sequestration of elevated levels of heavy-metal ions. These observations strongly suggest that the different forms of MT have different biological functions.


The analgesic effect of oxygen during percutaneous coronary intervention (the OXYPAIN Trial).
Oxygen is considered to have analgesic effects, but the evidence is weak. Oxygen may be harmful to the ischemic myocardium. The aim was to investigate the analgesic effect of oxygen during percutaneous coronary intervention (PCI) and to evaluate cardiac injury. The OXYPAIN was a phase II randomized trial with a double blind design. 305 patients were randomized to receive oxygen or atmospheric air during PCI. The patients were asked to score chest pain by the Visual-Analog Scale (VAS). The use of analgesic agents and troponin-t was measured. There was no significant difference in pain between the groups: oxygen: 2.0, [2.0-4.0], air: 2.0, [2.0-5.0] (median, interquartile range: 25-75%, P = 0.12). The median difference in score of VAS was [95% CI]: 0, [0-1.0]. The oxygen group received 0.44 ± 0.11 mg of morphine versus 0.46 ± 0.13, P = n.s. The peak value of troponin-t post-PCI was 38, [11-352] nmol/ml in the oxygen group and 61, [16-241] for patients treated with air, P = 0.46. The use of oxygen during PCI did not demonstrate any analgesic effect. There was no difference in myocardial injury measured with troponin-t or in the morphine dose. Our results do not support routine use of oxygen. (NCT01413841.).


Spontaneous mutations in the Big Blue transgenic system are primarily mouse derived.
The Big Blue transgenic mouse mutation detection system provides a powerful approach for measuring spontaneous and induced mutations in vivo. The observed mutations may contain a fraction of ex vivo or prokaryotic mutational events. Indeed, a modified, selectable form of the Big Blue assay seem to generate artifactual mutants under certain circumstances. Herein we review the evidence that circular mutants (i.e., the plaque circumference is at least 50% blue) collected in the standard Big Blue assay are derived primarily from the mouse. The most direct evidence is the similarity in the types of mutations found in jackpot and nonjackpot mutations. In addition, about half of the spontaneous mutations in the lacI transgene are transitions and transversions at CpG dinucleotides, a mammalian-specific feature. The mutation pattern observed at lacI is consistent with AT mutation pressure operating in a GC rich DNA and approaches that reported for observed germline human factor IX mutations. Furthermore, the spontaneous mutation pattern of circular Big Blue mutants differs significantly from that of an endogenous lacI gene in E. coli. Pinpoint mutants (a dot of blue color peripherally located in a wild type plaque), which a priori were not expected to be mouse-derived, have a mutation pattern consistent with the mutation pattern of an endogenous E. coli lacI gene. Analysis of induced mutagenesis studies reveals mutation frequencies and patterns for the Big Blue circular mutants which are comparable to endogenous genes. In reconstruction experiments, blue plaques derived from a superinfection with wild type and mutant phage produced approximately 50% blue and 50% clear plaques on replating. This phenomenon has not been seen when plaques derived from mouse were replated in the Big Blue assay. Collectively, the evidence strongly supports a murine origin for circular mutants recovered in the standard Big Blue assay. Validation of current assays is an essential step in determining the frequency and pattern of spontaneous murine-specific mutations. Defining this benchmark will be helpful in evaluating the next generation of transgenic mutation detection systems.


Morphologically Tunable Supramolecular Rectangular Microsheet and Microsaw Constructed by Hierarchical Self-Assembly Based on Hydrogen Bonds.
An amino acid derivative, thiophene (TDAV), as new building blocks for 2D supramolecular assembly is designed. Various square and rectangular microsheets are achieved and the aspect ratios are precisely regulated by controlling the polarity of cosolvent or water content. By the introduction of chirality, the novel microsaw is also achieved. It provides a new approach to prepare various kinds of unique supramolecular 2D materials with controllable shapes and sizes for future biological applications.


Endometrial carcinoma located in the right septate uterus cavity: a case report.
Endometrial cancer in patients with uterine congenital malformations is exceptional and there are only a few rare cases published in the literature. We report the case of a 67 years-old patient with an endometrial cancer located in the right cavity of a complete septate uterus.


Paradoxical development of pleural-based masses in patients with pleural tuberculosis during treatment: a clinical observational study in China.
To explored the clinical, pathological, and bacteriological characteristics of pleural-based masses occurred during anti-tuberculosis (TB) treatment in patients with pleural TB. Patients referred with newly diagnosed pleural TB were prospectively enrolled into the study. Patients were followed up throughout the treatment, and clinical data were recorded. Percutaneous biopsy and surgical tissues from pleural-based masses were examined histologically and samples sent for PCR. Cytokines in the pleural effusions and clinical factors were collected and compared between different patients. A total of 122 patients with pleural TB were enrolled, and 34.4% (42/122) displayed newly observed pleural-based mass during the treatment. Twelve cases underwent surgical resection at the 12 ± 0.5 months during the treatment course. Based on the surgical observation, 58.3% (7 /12) were located in pleura, 41.7% (5/12) were located in the lung parenchyma. Pathological observations showed that the pleural-based masses were typed as granulomatous inflammation, fibrous hyperplasia and necrosis. Mycobacterium tuberculosis PCR was positive in 57.1% of the cases (24/42). Any first-line anti-TB drug resistance gene mutations were positive in only 9.5% (4/42). Aside from 12 cases who underwent the surgical operation, 86.7% of the patients (26/30) still had a pleural-based mass at the end of 12 months treatment course. Patients with a pleural-based mass were younger, had a thicker pleural, a higher proportion of pleural adhesive, loculated pleural effusion and residual pleural effusion, and a higher level of LDH, ADA and lower glucose in pleural effusion than those without a pleural-based mass occurrence during the treatment (all Pcorr < 0.05). Pleural-based masses were observed in about one-third of patients with pleural TB. The masses were in the lung or pleura and were divided into three pathological types.


Antibody-induced changes in levels of cyclic adenosine monophosphate in leukaemic lymphocytes.
When L2C leukaemic B lymphocytes from guinea-pigs were incubated in vitro with antibody directed to their surface immunoglobulin (Ig), a rapid rise in intracellular adenosine 3':5'-phosphate (cyclic adenosine monophosphate, cAMP) was observed. Estimation of cAMP was by a protein-binding assay using bovine adrenal protein kinase. Increases up to 30-fold occurred within 30 seconds of incubation at 37 degrees C, to be succeeded by a fall which reached the basal level between 5 and 7 min. The response was proportional to the amount of antibody present. Cross-linking of surface Ig by the antibody was necessary, bivalent (Fab'gamma)2 from the antibody gave a rise in cAMP similar to that given by the parent molecule, whereas monomeric Fab'gamma was ineffective unless it was subsequently cross-linked by anti-antibody. The rise was too rapid to have required capping of the surface Ig for its induction. Not all perturbations of the plasma membrane by antibody induce such a surge in cAMP, since anti-beta2 microglobulin, also reacting with the lymphocyte surface, failed to alter cAMP concentration. The results emphasize that immunotherapy can be influenced by antibody altering the metabolic activity of target cells, quite apart from activation of immunological cytotoxic pathways.


Pathways to community transmission of COVID-19 due to rapid evaporation of respiratory virulets.
The formation of virus-laden colloidal respiratory microdroplets - the sneeze or cough virulets and their evaporation driven miniaturization in the open air are found to have a significant impact on the community transmission of COVID-19 pandemic. We simulate the motions and trajectories of virulets by employing laminar fluid flow coupled with droplet tracing physics. A force field analysis has been included considering the gravity, drag, and inertial forces to unleash some of the finer features of virulet trajectories leading to the droplet and airborne transmissions of the virus. Furthermore, an analytical model corroborates temperature (T) and relative humidity (RH) controlled droplet miniaturization. The study elucidates that the tiny (1-50 µm) and intermediate (60-100 µm) size ranged virulets tend to form bioaerosol and facilitate an airborne transmission while the virulets of larger dimensions (300 to 500 µm) are more prone to gravity dominated droplet transmission. Subsequently, the mapping between the T and RH guided miniaturization of virulets with the COVID-19 cases for six different cities across the globe justifies the significant contribution of miniaturization-based bioaerosol formation for community transmission of the pandemic.


Allobacillus salarius sp. nov., and Allobacillus saliphilus sp. nov., isolated from shrimp paste (ka-pi) in Thailand.
Two strains of moderately halophilic, Gram-stain-positive and spore-forming rods, designated as SKP4-8T and SKP8-2T isolated from shrimp paste (Ka-pi), were taxonomic studied based on polyphasic approach. Strain SKP4-8T grew at pH 6.0-9.0 (optimum 7.0), at 25-45 °C (optimum 37 °C) and in 1-16% (w/v) NaCl (optimum 5-10%). Strain SKP8-2T grew at pH 6.0-9.0 (optimum 8.0), at 25-45 °C (optimum 37 °C) and in 0-20% (w/v) NaCl (optimum 3-10%). The strains contained meso-diaminopimelic acid in cell-wall peptidoglycan and the major menaquinone was MK-7. Strain SKP4-8T contained iso-C15:0, anteiso-C15:0 and iso-C17:0; and strain SKP8-2T contained anteiso-C15:0, iso-C15:0, iso-C16:0 and antesio-C17:0 as major cellular fatty acids. Phosphatidylglycerol, diphosphatidylglycerol, unknown phospholipids and an unknown glycolipid were detected as major polar lipids. On the basis of 16S rRNA gene sequence analysis, strains SKP4-8T and SKP8-2T belonged to the genus Allobacillus and were closely related to Allobacillus halotolerans LMG 24826T with 98.8% and 99.3% similarity, respectively. The comparative genome analysis based on average nucleotide identity (ANI) and digital DNA-DNA hybridization revealed that both strains showed the values below 95 and 70%, from each other and from Allobacillus halotolerans LMG 24826T, respectively. Based on the data from this polyphasic study, strains SKP4-8T and SKP8-2T represent novel species of the genus Allobacillus and the name Allobacillus salarius sp. nov. was proposed for SKP4-8T (= KCTC 33905T = LMG 30016T = TISTR 2499T); and Allobacillus saliphilus sp. nov. for SKP8-2T (= KCTC 33906T = LMG 29682T = TISTR 2558T).


CD229 (Ly9) a Novel Biomarker for B-Cell Malignancies and Multiple Myeloma.
CD229 (Ly9) homophilic receptor, which belongs to the SLAM family of cell-surface molecules, is predominantly expressed on B and T cells. It acts as a signaling molecule, regulating lymphocyte homoeostasis and activation. Studies of CD229 function indicate that this receptor functions as a regulator of the development of marginal-zone B cells and other innate-like T and B lymphocytes. The expression on leukemias and lymphomas remains poorly understood due to the lack of CD229 monoclonal antibodies (mAb) for immunohistochemistry application (IHC). In this study, we used a new mAb against the cytoplasmic region of CD229 to study the expression of CD229 on normal tissues and B-cell malignancies, including multiple myeloma (MM), using tissue microarrays. We showed CD229 to be restricted to hematopoietic cells. It was strongly expressed in all cases of MM and in most marginal-zone lymphomas (MZL). Moderate CD229 expression was also found in chronic lymphocyte leukemia (CLL), follicular (FL), classic mantle-cell (MCL) and diffuse large B-cell lymphoma. Given the high expression on myeloma cells, we also analyzed for the presence of soluble CD229 in the sera of these patients. Serum levels of soluble CD229 (sCD229) at the time of diagnosis in MM patients could be useful as a prognostic biomarker. In conclusion, our results indicate that CD229 represents not only a useful biomarker but also an attractive therapeutic target.


Model spread in tropical low cloud feedback tied to overturning circulation response to warming.
Among models participating in the Coupled Model Intercomparison Project phase 6 (CMIP6), here we show that the magnitude of the tropical low cloud feedback, which contributes considerably to uncertainty in estimates of climate sensitivity, is intimately linked to tropical deep convection and its effects on the tropical atmospheric overturning circulation. First, a reduction in tropical ascent area and an increased frequency of heavy precipitation result in high cloud reduction and upper-tropospheric drying, which increases longwave cooling and reduces subsidence weakening, favoring low cloud reduction (Radiation-Subsidence Pathway). Second, increased longwave cooling decreases tropospheric stability, which also reduces subsidence weakening and low cloudiness (Stability-Subsidence Pathway). In summary, greater high cloud reduction and upper-tropospheric drying (negative longwave feedback) lead to a more positive cloud feedback among CMIP6 models by contributing to a greater reduction in low cloudiness (positive shortwave feedback). Varying strengths of the two pathways contribute considerably to the intermodel spread in climate sensitivity.


molecular mechanism of lysidine synthesis that determines tRNA identity and codon recognition.
Lysidine (2-lysyl cytidine) is a lysine-containing cytidine derivative commonly found at the wobble position of bacterial AUA codon-specific tRNA(Ile). This modification determines both codon and amino acid specificities of tRNA(Ile). We previously identified tRNA(Ile)-lysidine synthetase (tilS) that synthesizes lysidine, for which it utilizes ATP and lysine as substrates. Here, we show that lysidine synthesis consists of two consecutive reactions that involve an adenylated tRNA intermediate. A mutation study revealed that Escherichia coli TilS discriminates tRNA(Ile) from the structurally similar tRNA(Met) having the same anticodon loop by recognizing the anticodon loop, the anticodon stem, and the acceptor stem. TilS was shown to bind to the anticodon region and 3' side of the acceptor stem, which cover the recognition sites. These findings reveal a dedicated mechanism embedded in tRNA(Ile) that controls its recognition and discrimination by TilS, and indicate the significance of this enzyme in the proper deciphering of genetic information.


Experimental Reproduction of Dynamic Fluctuation of TSH Receptor-Binding Antibodies Between Stimulation and Inhibition.
Hyperthyroidism in Graves disease (GD) is caused by autoantibody stimulation of the TSH receptor (TSHR). TSHR autoantibody (TSHR-Ab) activity is measured routinely by inhibition of labeled ligand (TSH or M22) binding to the TSHR [TSH-binding inhibitory immunoglobulins (TBIIs)] or by stimulation of cAMP production in isolated cells [TSH receptor-stimulating antibodies (TSAbs)]. Usually, measurements of TSHR-Abs by TBIIs agree reasonably well with TSAb values at least in the setting of hyperthyroidism, and both measurements tend to change in parallel during treatment with some exceptions. In this study, we describe three unusual cases, which illustrate nearly pure stimulating, blocking, or neutral properties of TSHR-Abs. Whether patient serum TSHR-Abs can be reproduced by mixtures of human monoclonal autoantibodies to the TSHR was studied because the sera in most patients show moderate properties having both of TBII and TSAb activities. We compared the TBII and TSAb activities of serum from four unusual patients in detail with mixtures of human monoclonal TSHR-Abs (mAbs) M22 (stimulating), K1-18 (stimulating), and K1-70 (blocking). Characteristic of a patient's serum was similar to M22 or K1-18, another was similar to K1-70, whereas another was similar to a mixture of K1-70 and M22 (or K1-18). Additionally, some patients seemed to have neutral TSHR-Abs in their sera. Our studies suggest that the characteristics of TSHR-Abs in the patient serum can be mimicked by mixtures of human mAbs to the TSHR, stimulating, blocking, and neutral if any.


Serum glycoproteins in the liver diseases. VIII. Desialylated glycoproteins in the liver cirrhosis.
Serum desialylated glycoprotein livel of cirrhotic patients was determined and a diagnostically significant elevation of these proteins was observed. The level of these patients was usually 2--10 times of that seen in normal subjects and the elevation was signifi-ant (p less than 0.001) when compared to the level in patients with chronic aggressive hepatitis, severe (2B). Serial determinations of these proteins in the cirrhotic patients showed no correlation between them and SGPT as a whole but in several cases in which SGPT fluctuated the former associated with the latter. In patients with decompensated cirrhotic liver these proteins returned nearly to the level of compensated patients when it was improved. The level of these proteins in cirrhotic patients correlated, not always, with serum albumin (r = -0.46, p less than 0.02) and indocyanine green clearance rate (r = -0.73, p less than 0.05), but not with SGPT as well as the other liver function tests.


Strain, age, but not gender, influence ulcer severity induced by water-restraint stress.
Two ulcerogenic procedures, supine restraint (SR) and water restraint (WR) were compared. In Experiment 1, Fischer-344 (F344), Sprague-Dawley (S-D), Wistar, Long-Evans (L-E), Spontaneously Hypertensive rats (SHR) and Wistar Kyoto normotensive (WKY) rats were exposed to SR and WR. WR produced more ulcers than SR. There was no difference in ulcer scores between WKY, F344 and L-E but these rats had significantly more ulcers as compared to SHR, Wistar and S-D rats. In Experiment 2, 4- and 16-month-old SHR, WKY and F344 rats were exposed to SR and WR. The older WKY rats had more ulcers than all other treatment groups. Experiment 3 revealed no significant differences between male and female rats exposed to either SR or WR. Body temperature (BT) scores obtained after restraint and after 2-hr postrestraint rest were only marginally related to ulcer severity. Rats exposed to WR had lower BT scores but the strain and age ulcer differences did not have corresponding BT differences. These studies revealed the following: the ulcer susceptibility of WKY rats; the WR technique is a useful ulcerogenic procedure; and hypothermia is a weak covariant to restraint-induced stress ulcer.


Nuclear granules recognized by some monoclonal antibodies against intermediate filament protein locate on chromosomes during mitosis.
AC54 monoclonal antibody (MAb), an anti-desmin MAb, recognizes both intermediate filaments (IFs) and nuclear granules in BHK21/C13 cells. To investigate nuclear granules, similar MAbs were obtained by using desmin fraction as an antigen. Among them, DSB389 MAb recognized mainly nuclear granules in HeLa and rat liver cells. The nuclear granules in HeLa cells were aligned in arrays, sometimes connected by, or part of, a rope-like structure, and stable against treatment with 0.5% Triton X-100 and 2 M NaCl. They located on or around the chromosomes during mitosis. Essentially the same results were obtained with DSB860 and AC54 MAbs. The distribution of the granules in liver nuclei recognized by DSB389 MAb was similar to that of DNA and was different from that of the nuclear pore complexes. The biological significance of the nuclear granules is discussed.