Cryopreservation of alginate-fibrin beads involving bone marrow derived mesenchymal stromal cells by vitrification.
Application of cell--biomaterial systems in regenerative medicine can be facilitated by their successful low temperature preservation. Vitrification, which avoids ice crystal formation by amorphous solidification, is an emerging approach to cryopreservation. Developing vitrification strategy, effective cryopreservation of alginate-fibrin beads with porcine mesenchymal stromal cells has been achieved in this study. The cell-biomaterial constructs were pre-cultured for 20 days before cryopreservation, allowing for cell proliferation and construct stabilization. Ethylene glycol (EG) was employed as the basic cryoprotectant for two equilibration solutions. Successful cryopreservation of the constructs was achieved using vitrification solution composed of penetrating (EG MW 62 Da) and non-penetrating (sucrose MW 342 Da) cryoprotectants. Stepwise procedure of introduction to and removal of cryoprotectants was brief; direct plunging into liquid nitrogen was applied. Cell viability, evaluated by combining live/death staining and confocal laser microscopy, was similar for both control and vitrified cells in the beads. No detectable damage of microstructure of cryopreserved beads was found as shown by scanning electron microscopy. Both osteogenically induced control and vitrified cells in the constructs were equally capable of mineral production and deposition. There was no statistically significant difference in metabolic activity and proliferation between both groups during the entire culture period. Our study leads to the conclusion that the developed cryopreservation protocol allowed to maintain the integrity of the beads while preserving the ability of the pig bone marrow derived mesenchymal stromal cells to proliferate and subsequently differentiate; demonstrating that vitrification is a promising approach for cryopreservation of "ready-to-use" cell-biomaterial constructs.


Electrophysiological patterns of oropharyngeal swallowing in multiple sclerosis.
We performed an electrophysiological study of swallowing (EPSS) in multiple sclerosis (MS) to describe oropharyngeal swallowing abnormalities and to analyze their correlations with dysphagia and with overall neurological impairment. Neurological examinations were quantified using the Kurtzke Functional Systems and the Expanded Disability Status Scale (EDSS). Dysphagia was evaluated using the Dysphagia in Multiple Sclerosis (DYMUS) questionnaire, while fiberoptic endoscopic evaluation of swallowing (FEES) was used to establish the degree of aspiration and penetration, graded using the penetration-aspiration scale (PAS). The EPSS measured the duration of suprahyoid/submental muscle EMG activity (SHEMG-D), the duration of the laryngeal-pharyngeal mechanogram (LPM-D), and the duration of the pause in cricopharyngeal muscle EMG activity (CPEMG-PD); it also measured the interval between onset of the suprahyoid/submental muscle EMG activity (SHEMG) and onset of the laryngeal-pharyngeal mechanogram (I-SHEMG-LPM). 92% of patients showed at least one electrophysiological abnormality. I-SHEMG-LPM correlated positively with the DYMUS questionnaire. I-SHEMG-LPM, SHEMG-D, and DYMUS correlated positively with the PAS. Moderate to severe bladder sphincter dysfunction was associated with a significant reduction, or absence, of CPEMG-PD. EPSS improves our understanding of the pathophysiology of dysphagia in MS. This investigation could be useful in MS patients with swallowing abnormalities.


Pigmented preretinal membranes.
Continuous growth and an increase in pigmentation of a preretinal membrane occurred in a 46-year-old woman after a scleral buckling procedure for rhegmatogenous retinal detachment. Ten additional patients had pigmented preretinal membranes in eyes treated for rhegmatogenous retinal detachment. Although we made no attempts to determine the incidence of pigmented preretinal membrane formation in eyes with retinal detachments, these 11 cases, in a consecutive series of 500 eyes treated for rhegmatogenous retinal detachment, indicated that the development of pigmented preretinal membranes is not uncommon. Our clinical observations supported recent experimental findings implicating retinal pigment epithelial cells as a cause of preretinal membrane formation.


Brown adipose tissue but not tibia exhibits a dramatic response to acute reduction in environmental temperature in growing male mice.
Mice are typically housed at room temperature (∼22 °C), which is well below their thermoneutral zone and results in cold stress. Chronic cold stress leads to increased adaptive thermogenesis and reductions in cancellous bone volume and bone marrow adipose tissue mass in long bones of growing mice. There is strong evidence that increased neuronal activity initiates the metabolic response of intrascapular brown adipose tissue (BAT) to cold stress, but it is less clear whether bone is regulated through a similar mechanism. Therefore, we compared the short-term response of BAT and whole tibia to a reduction in environmental temperature. To accomplish this, we transferred a group of 6-week-old male mice from 32 °C to 22 °C housing and sacrificed the mice 24 h later. Age-matched controls were maintained at 32 °C. We then evaluated expression levels of a panel of genes related to adipocyte differentiation and fat metabolism in BAT and tibia, and a panel of genes related to bone metabolism in tibia. The decrease in housing temperature resulted in changes in expression levels for 47/86 genes related to adipocyte differentiation and fat metabolism in BAT, including 9-fold and 17-fold increases in Ucp1 and Dio2, respectively. In contrast, only 1/86 genes related to adipocyte differentiation and fat metabolism and 4/84 genes related to bone metabolism were differentially expressed in tibia. These findings suggest that bone, although innervated with sensory and sympathetic neurons, does not respond as rapidly as BAT to changes in environmental temperature.


High-energy phosphate compounds and some glycolytic substrates in the rat brain during hypoxia.
The influence of acute and prolonged (with resuscitation) hypoxia, upon some glycolytic substrates and high energy phosphate compounds in the rat brain, was studies. Acute hypoxia decreased the content of glucose and glucose-6-phosphate as well as of high energy phosphate compounds. Stimultaneously, an increase of lactic level was recorded. Some adaptation features could be observed, when 30-minute hypoxia with resuscitation was studied. Brain content of high energy compounds returned to the normal value, while that of glucose increased twice over control. However, the level of lactic acid was still increased.


Tailored flow sequestration treatment using high-flow and low-flow bypass for partially thrombosed giant internal carotid artery aneurysm-a technical case report.
Direct clipping of giant partially thrombosed intracranial internal carotid artery (ICA) aneurysms is challenging, especially when important perforating arteries are involved. Proximal occlusion with bypass represents a possible alternative approach. An 80-year-old female presented with worsening visual acuity and severe headache caused by partially thrombosed giant (38 mm in diameter) aneurysms of the right ICA, suggestive of impending rupture. Direct clipping in conjunction with temporary occlusion of the lesion involving the anterior choroidal artery (AChA) was considered too risky. Thus, we sequestrated the ipsilateral ICA flow into a low-flow and a high-flow system using two external carotid artery (ECA)-ICA bypasses and one in situ bypass with cervical ICA ligation. As a result, the low-flow system by the superficial temporal artery-middle cerebral artery (MCA) bypass perfused mainly the proximal MCA lesions and aneurysm, whereas the high-flow system by ECA-radial artery-M2 bypass exclusively supplied the residual distal MCA area. This tailored flow sequestration successfully interrupted intra-aneurysmal flow and accelerated near-complete thrombosis of the aneurysm while preserving the AChA and avoiding any significant neurological deterioration. We conclude that this method is effective for the management of giant partially thrombosed aneurysms of the ICA, especially when direct clipping is difficult.


Correction to: Perioperative Management of Elderly patients (PriME): recommendations from an Italian intersociety consensus.
The article Perioperative Management of Elderly patients (PriME).


Associations of active-duty mental health trajectories with post-military adjustment: Results from the STARRS Longitudinal Study.
Many servicemembers experience difficulties transitioning from military to civilian life. We examined whether changes in mental health observed during active duty were associated with indices of post-military adjustment. Survey data from the multi-wave Army STARRS Pre/Post Deployment Study (PPDS; conducted 2012-2014) were linked to follow-up data from wave 1 of the STARRS Longitudinal Study (STARRS-LS1; conducted 2016-2018). Empirical Bayes estimates of intercepts and slopes of posttraumatic stress, problematic anger, and depressive symptoms during the PPDS were extracted from mixed-effects growth models and evaluated as predictors of life stress among 1080 participants who had separated or retired from the Army at STARRS-LS1; and of job satisfaction among 586 veterans who were employed at STARRS-LS1. Higher average levels and larger increases in posttraumatic stress, anger, and depression over the deployment period were each associated with increased stress and (in the case of anger and depression) reduced job satisfaction. Posttraumatic stress and anger slopes were associated with overall stress (b = 5.60, p < 0.01 and b = 15.64, p = 0.04, respectively) and relationship stress (b = 5.50, p = 0.01 and b = 22.86, p = 0.01, respectively) beyond the average levels of those symptoms. Some transition-related difficulties may have resolved before outcome assessment; some measures were not previously validated. Larger increases in posttraumatic stress and anger over a deployment period were associated with increased stress after leaving the Army, even after controlling for average symptom levels during the same period. Monitoring changes in mental health during active duty may help identify personnel who need additional support to facilitate the military-to-civilian transition.


Cross-linking of CD81 by HCV-E2 protein inhibits human intrahepatic plasmacytoid dendritic cells response to CpG-ODN.
Plasmacytoid dendritic cells (pDCs) are reported to be defective in HCV-infected patients, the mechanisms of which remain poorly understood. We isolated liver derived mononuclear cells (LMNCs) and pDCs from normal liver tissues of benign tumor dissections and liver transplant donors. Isolated pDCs and LMNCs were cultured with precoated HCV envelop protein E2 (HCV-E2) or anti-CD81 mAb in the presence of CpG-ODN. Our results show that cross-linking of CD81 by either HCV-E2 or anti-CD81 mAb inhibits IFN-α secretion in CpG-induced pDCs; down-regulates HLA-DR, CD80 and CD86 expression in pDCs; and suppresses CpG-ODN induced proliferation and survival of pDCs. The blockade of CD81 by soluble anti-CD81 antibody restores pDCs response to CpG-ODN. These results suggest that HCV E2 protein interacts with CD81 to inhibit pDC maturation, activation, and IFN-α production, and may thereby contribute to the impaired innate anti-viral immune response in HCV infection.


Quantitative PCR analysis and protein distribution of drug transporter genes in the rat cochlea.
Membrane transporters can be major determinants in the targeting and effectiveness of pharmaceutical agents. A large number of biologically important membrane transporters have been identified and localized to a variety of tissues, organs and cell types. However, little is known about the expression of key membrane transporters in the inner ear, a promising site for targeted therapeutics, as well as a region vulnerable to adverse drug reactions and environmental factors. In this study, we examined the levels of endogenous membrane transporters in rat cochlea by targeted PCR array analysis of 84 transporter genes, followed by validation and localization in tissues by immunohistochemistry. Our studies indicate that several members of the SLC, VDAC and ABC membrane transporter families show high levels of expression, both at the RNA and protein levels in the rat cochlea. Identification and characterization of these membrane transporters in the inner ear have clinical implications for both therapeutic and cytotoxic mechanisms that may aid in the preservation of auditory function.


Inorganic Solar Cells Based on Electrospun ZnO Nanofibrous Networks and Electrodeposited Cu2O.
The nanostructured ZnO/copper oxide (Cu2O) photovoltaic devices based on electrospun ZnO nanofibrous network and electrodeposited Cu2O layer have been fabricated. The effects of the pH value of electrodeposition solution and the Cu2O layer thickness on the photovoltaic performances have been investigated. It is revealed that the pH value influences the morphology and structure of the Cu2O layer and thus the device performances. The Cu2O layer with an appropriate thickness benefits to charge transfer and light absorption. The device prepared at the optimal conditions shows the lowest recombination rate and exhibits a power conversion efficiency of ~0.77 %.


Unimolecular Reactivity of [Cu(R)(CF3 )3 ]- Complexes (R=Organyl): Stepwise versus Concerted Mechanism in Copper-Mediated Trifluoromethylation.
The cuprate complexes [Cu(R)(CF3 )3 ]- (R=organyl) offer an efficient synthetic access to valuable trifluoromethylation products RCF3 . Here, electrospray-ionization mass spectrometry is used to analyze the formation of these intermediates in solution and probe their fragmentation pathways in the gas phase. Furthermore, the potential energy surfaces of these systems are explored by quantum chemical calculations. Upon collisional activation, the [Cu(R)(CF3 )3 ]- complexes (R=Me, Et, Bu, s Bu, allyl) afford the product ions [Cu(CF3 )3 ]⋅- and [Cu(CF3 )2 ]- . The former obviously results from an R⋅ loss, whereas the latter originates either from the stepwise release of R⋅ and CF3 ⋅ radicals or a concerted reductive elimination of RCF3 . The gas-phase fragmentation experiments as well as the quantum chemical calculations indicate that the preference for the stepwise reaction toward [Cu(CF3 )2 ]- increases with the stability of the formed organyl radical R⋅. This finding suggests that the recombination of R⋅ and CF3 ⋅ radicals may possibly contribute to the formation of RCF3 from [Cu(R)(CF3 )3 ]- in synthetic applications. In contrast, the [Cu(R)(CF3 )3 ]- complexes (R=aryl) only yield [Cu(CF3 )2 ]- when subjected to collision-induced dissociation. These species exclusively undergo a concerted reductive elimination because the competing stepwise pathway is disfavored by the low stability of aryl radicals.


[The role of inositol triphosphate and diacylglycerol as intracellular second messengers].
The paper presents the mode of formation and action of inositol trisphosphate and diacylglycerol and their role as secondary messengers.


UTRdb 2.0: a comprehensive, expert curated catalog of eukaryotic mRNAs untranslated regions.
The 5' and 3' untranslated regions of eukaryotic mRNAs (UTRs) play crucial roles in the post-transcriptional regulation of gene expression through the modulation of nucleo-cytoplasmic mRNA transport, translation efficiency, subcellular localization, and message stability. Since 1996, we have developed and maintained UTRdb, a specialized database of UTR sequences. Here we present UTRdb 2.0, a major update of UTRdb featuring an extensive collection of eukaryotic 5' and 3' UTR sequences, including over 26 million entries from over 6 million genes and 573 species, enriched with a curated set of functional annotations. Annotations include CAGE tags and polyA signals to label the completeness of 5' and 3'UTRs, respectively. In addition, uORFs and IRES are annotated in 5'UTRs as well as experimentally validated miRNA targets in 3'UTRs. Further annotations include evolutionarily conserved blocks, Rfam motifs, ADAR-mediated RNA editing events, and m6A modifications. A web interface allowing a flexible selection and retrieval of specific subsets of UTRs, selected according to a combination of criteria, has been implemented which also provides comprehensive download facilities. UTRdb 2.0 is accessible at http://utrdb.cloud.ba.infn.it/utrdb/.


Progressive relaxation as a function of procedural variations and anxiety level.
Progressive relaxation is subject to many procedural variations. Although most investigators follow the 5-7 s tension interval advocated by Bernstein and Borkovec (1973), longer tension intervals may contribute to greater relaxation effects. This study compared tension intervals of 5, 15, and 45 s. Since the development of control over muscle tension requires attention to feedback signals, a focus on internal proprioceptive sensations was contrasted with an external condition in which the subjects silently subtracted numbers immediately following the tension-release cycle. Longer periods of tension resulted in lower EMG levels, with the 45-s tension interval producing the greatest relaxation. Those individuals who focused on physiological sensations during training were able to achieve greater levels of relaxation after the training phase was completed. There was a significant interaction between tension interval duration and focusing; subjects in the 45-s interval who focused on physiological functioning showed the greatest arousal reduction. Anxiety level did not differentially influence the effects of procedural variations in training. Clearly, additional parametric studies in relaxation training are needed.


Relationships between carrier-mediated transport of folate compounds by L1210 leukemia cells: evidence for multiplicity of entry routes with different kinetic properties expressed in plasma membrane vesicles.
Various independent kinetic criteria for indicating multiplicity of carrier-mediated entry of folate compounds into L1210 cell plasma membrane vesicles are studied. We find a marked inconsistency between values for influx Km and Ki in reciprocal experiments measuring competition between various folate compounds as well as inconsistent effects of transloading shown for 5-formyltetrahydrofolate influx, but not folic acid influx. These results argue strongly against a one-carrier model for transport of folate compounds. The most straightforward interpretation of our data is that two distinct transport systems mediate entry of folate compounds in L1210 plasma membrane vesicles. If a two-carrier model is correct, then our data indicate that one of the carriers has low capacity and high affinity for folate coenzymes and methotrexate. This system is apparently negligible as a transport route for folic acid. Transtimulation of initial influx by substrates of the low capacity system is obtained following transloading with coenzymes but not by transloading with folic acid. Our data indicate that the second folate transport system postulated by the two-carrier model has a low affinity for all the folate compounds studied. Nevertheless, the putative second system is significant, especially for folic acid transport, because it has a much higher capacity than the first transport system. In contrast to the first system, transloading with any of the folate compounds studied had no effect on initial influx mediated by the second folate transport system. The two systems are also differentially inhibited by pCMBS, DIDS and SITS and the influx Vmax for the high-affinity/low-capacity system was altered in a vesicle preparation derived from a methotrexate resistant L1210 cell line.


Acute caffeine intake increases performance in the 15-s Wingate test during the menstrual cycle.
In male athletes, caffeine is considered an ergogenic aid to increase anaerobic performance during the Wingate anaerobic test (WANT). However, information about the effect of caffeine on WANT performance in female athletes is contradictory. Furthermore, it is unknown whether the ergogenicity of caffeine is present during all the phases of the menstrual cycle. The aim of this study was to investigate the effects of caffeine intake on WANT performance during 3 phases of the menstrual cycle. Thirteen well-trained eumenorrhoeic triathletes participated in a double-blind, placebo-controlled, cross-over experimental trial. On 2 different days in each phase, and in randomized order, participants ingested caffeine (3 mg kg-1 ) or a placebo (cellulose). The menstrual cycle phases were individually characterized as follows: (i) early follicular; (ii) preovulatory; and (iii) midluteal. In each trial, participants performed a 15-s adapted version of the WANT. In comparison to the placebo, caffeine increased peak power during the WANT in the early follicular (8.6 ± 0.8 vs 8.9 ± 0.9 W/kg, P = .04; effect size [d] = 0.45), preovulatory (8.6 ± 0.9 vs 8.9 ± 0.9 W/kg, P = .04; d = 0.23) and mid-luteal phases (8.6 ± 0.8 vs 8.9 ± 0.9 W/kg, P < .01; d = 0.52). The ergogenic effect of caffeine on WANT peak cycling power was of a similar magnitude in the follicular, preovulatory, and mid-luteal phases. These results suggest that caffeine increases performance in the 15-s Wingate test in women athletes and it might be considered an ergogenic aid to increase anaerobic performance in eumenorrhoeic women during their menstrual cycle.


A computational model for learning from repeated traumatic experiences under uncertainty.
Traumatic events can lead to lifelong, inflexible adaptations in threat perception and behavior, which characterize posttraumatic stress disorder (PTSD). This process involves associations between sensory cues and internal states of threat and then generalization of the threat responses to previously neutral cues. However, most formulations neglect adaptations to threat that are not specific to those associations. To incorporate nonassociative responses to threat, we propose a computational theory of PTSD based on adaptation to the frequency of traumatic events by using a reinforcement learning momentum model. Recent threat prediction errors generate momentum that influences subsequent threat perception in novel contexts. This model fits primary data acquired from a mouse model of PTSD, in which unpredictable footshocks in one context accelerate threat learning in a novel context. The theory is consistent with epidemiological data that show that PTSD incidence increases with the number of traumatic events, as well as the disproportionate impact of early life trauma. Because the theory proposes that PTSD relates to the average of recent threat prediction errors rather than the strength of a specific association, it makes novel predictions for the treatment of PTSD.


Swainsonine, an inhibitor of glycoprotein processing, enhances cytotoxicity of large granular lymphocytes.
In the present study we investigated the effects of inhibitors of glycoprotein processing on cytotoxicity of human large granular lymphocytes (LGL). The incubation of LGL for 36 h with 0.5 microgram/ml swainsonine (SW), which is an inhibitor of mannosidase II, resulted in the augmentation of cytotoxicity of LGL against an NK-resistant colon carcinoma cell line (Colo-320DM) without increase of binding frequency of LGL to target cells or of cell proliferation. The enhanced cytotoxicity was associated with increased binding of concanavalin A to SW-treated LGL. The augmentation of cytotoxicity was also seen by 1-deoxymannojirimycin (1-DMN), an inhibitor of mannosidase I, but much higher amounts of this agent were needed to get the same level of augmentation as that with SW. Other inhibitors of glycoprotein processing such as castanospermine and 1-deoxynojirimycin (1-DN) did not show any augmentative effects on LGL cytotoxicity. The enhancement of cytotoxicity by SW was abolished by the addition of rabbit anti-human interleukin (IL-2) antibody to the culture. This result suggests that IL-2 is involved in the augmentation of cytotoxicity of LGL by SW. The presence of SW in the culture of LGL together with IL-2 also enhanced LAK generation compared to that with IL-2 alone. Thus, our results suggest that SW should be recognized as an efficient immunopotentiator and that modulation of carbohydrate moieties elicited by SW may shed further light on the mechanism of LGL activation.


Obesity Modulates Inflammation and Lipid Metabolism Oocyte Gene Expression: A Single-Cell Transcriptome Perspective.
It is hypothesized that obesity adversely affects the ovarian environment, which can disrupt oocyte maturation and embryonic development. This study aimed to compare oocyte gene expression profiles and follicular fluid (FF) content from overweight/obese (OW) women and normal-weight (NW) women who were undergoing fertility treatments. Using single-cell transcriptomic analyses, we investigated oocyte gene expression using RNA sequencing. Eleven OW women and 13 NW women undergoing fertility treatments were enrolled. Oocyte messenger RNA profiles as well as serum and FF hormone and lipid levels were assessed. OW women had significantly higher body mass index, body fat percentage, and serum homeostatic model assessment-insulin resistance index compared with NW women (P < 0.01). Serum leptin and C-reactive protein (CRP) levels as well as FF leptin, CRP, and triglyceride levels were increased (P < 0.05) in OW compared with NW women. Oocytes from OW women had increased expression of proinflammatory (CXCL2; P = 0.071) and oxidative stress-related (DUSP1; P = 0.051) genes but had decreased expression of GAS7 (fat metabolism; P = 0.065), TXNIP (oxidative stress; P = 0.055), and transcription factors ID3 (P = 0.075) and TWIST1 (P = 0.099) compared with NW women. These findings provide evidence for the significant influence of body composition on oocyte transcript abundance in women undergoing hormonal induction to retrieve oocytes. They further identify the potential for maternal diet to influence oocyte gene expression. The preconception period is, therefore, an important window of opportunity to consider for lifestyle interventions.


[Camillo Golgi: historical profile of a Nobel Prize politically engaged].
The article analyzes the role and activity of Camillo Golgi as Senator of the Italian Parliament in the light of the official acts which testify his modalities of political intervention, his style and his proposals, with the aim of verifying if and how his experience as scientific researcher and teacher at the University of Pavia shaped his political life.


Autotransfusion in the emergency department of a community hospital.
The Bentley-100-Autotransfusion System was used in eight cases in the emergency department of St. Francis Hospital of Lynwood. The majority of the patients sustained massive chest trauma and the amount of blood salvaged ranged from 1,600 to 14,000 cc. Although the clotting factors and platelet counts altered greatly during the first postoperative days, they returned to normal within 72 hours. The advantages, disadvantages, and controversial areas in autotransfusion are discussed.


Melatonin Ameliorates the Progression of Alzheimer's Disease by Inducing TFEB Nuclear Translocation, Promoting Mitophagy, and Regulating NLRP3 Inflammasome Activity.
The NLRP3 inflammasome is overactivated in the brains of APP/PS1 transgenic mice and AD patients, and mitophagy has an obvious negative regulatory role on NLRP3 inflammasome activation. The protective effect of melatonin in AD may be related to the regulation of mitophagy and NLRP3 inflammasome activity. TFEB plays a critical role in maintaining autophagy/mitophagy. Studies have found that TFEB plays a protective role in AD. APP/PS1 transgenic mice were given melatonin in their drinking water for 3 months. Compared with mice without melatonin treatment, the mice given melatonin showed changes in the following features: (1) cognitive function, (2) mitophagy-related proteins in the brain, (3) ROS, (4) NLRP3 inflammasome and related proteins and the concentrations of inflammatory cytokines, and (5) Aβ deposition. In in vitro experiments, effects of melatonin on mitophagy, NLRP3 inflammasome activity, and TFEB in SH-SY5Y cells with Aβ 25-35 were observed. TFEB knockdown was implemented in combination with Aβ 25-35 and melatonin treatment, and the expressions of TFEB, Parkin, p62, IL-1β, caspase-1, ROS, and IL-18 were explored. Melatonin improved cognitive function in APP/PS1 transgenic mice and decreased ROS and senile plaques. Melatonin promoted mitophagy in SH-SY5Y cells with Aβ 25-35 and APP/PS1 transgenic mice. NLRP3 inflammasome activity was inhibited, and the concentrations of IL-18 and IL-1βwere clearly reduced. Compared with C57/BL6J mice, the amount of TFEB in the brain nucleus of APP/PS1 transgenic mice was decreased. Melatonin treatment increased the nuclear translocation of TFEB in SH-SY5Y cells. TFEB knockout was implemented in combination with Aβ 25-35 and MT treatment; the expressions of Parkin, p62, caspase-1, IL-1β, IL-18, and ROS were accelerated. Melatonin promotes mitophagy by inducing TFEB nuclear translocation, downregulates NLRP3 inflammasome activation, and exerts protective effects in SH-SY5Y cells and APP/PS1 transgenic mice.


Experimental fractures of the mandibular body of sheep and dogs. A new technique.
In order to create artificial fractures in animals resembling those which occur naturally, a new technique for fracture production was developed. Using a specially designed clamp, artificial fractures were produced in the mandibular body of seven sheep and 12 dogs. Radiographical analysis of the effects of the clamp on the mandibular body showed simple, single vertical fractures in the buccal cortex, inferior border and the alveolar process comparable to those which occur naturally. Using the technique described it was possible, in all cases, to carry out treatment with the use of 4-hole poly(L-lactide) plates and screws.


Metals in Alzheimer's and Parkinson's diseases.
There has been steadily growing interest in the participation of metal ions (especially, zinc, copper, and iron) in neurobiological processes, such as the regulation of synaptic transmission. Recent descriptions of the release of zinc and copper in the cortical glutamatergic synapse, and influencing the response of the NMDA receptor underscore the relevance of understanding the inorganic milieu of the synapse to neuroscience. Additionally, major neurodegenerative disorders, including Alzheimer's disease and Parkinson's disease, are characterized by elevated tissue iron, and miscompartmentalization of copper and zinc (e.g. accumulation in amyloid). Increasingly sophisticated medicinal chemistry approaches, which correct these metal abnormalities without causing systemic disturbance of these essential minerals, are being tested. These small molecules show promise of being disease-modifying.


Treatment needs and predictive capacity of explanatory variables of oral disease in young athletes with an intellectual disability in Europe and Eurasia.
To evaluate the oral condition and treatment needs of young athletes with intellectual disability (ID) from 53 countries of Europe and Eurasia who participated in the Special Olympics European Games held in Antwerp, October 2014. A cross- sectional study was undertaken with data collected through standardised procedures from consenting athletes under 21 years of age. Oral hygiene habits, reports of oral pain and presence of gingival signs, sealants, untreated caries and missing teeth were recorded. Data analysis was performed in SPSS to produce descriptive statistics and explanatory variables for untreated decay, and gingival signs of disease were tested with Multilevel Generalized Linear Mixed Models. Five hundred three athletes participated in this study (mean age 17 yrs). Untreated decay was recorded in 33.4% of the participants and 38.7% of them had signs of gingival disease. Absence of untreated decay was associated with lower chances of gingival signs, while absence of sealants was related with higher chances of untreated decay. There is consistent evidence of persistent need for increased promotion of oral health, as well as preventive and restorative treatment in young athletes with ID in Europe and Eurasia. Due to the limited predictive capacity of the studied variables for oral disease, further studies including other related factors are needed.


TDDFT study on the sensing mechanism of a fluorescent sensor for fluoride anion: Inhibition of the ESPT process.
The fluoride-sensing mechanism of a reported salicylaldehyde-based sensor (J. Photochem. Photobiol. B 2014, 138, 75) has been investigated by the TDDFT method. The present theoretical study indicates that there is an excited-state proton transfer (ESPT) process from the phenolic O-H moiety to the neighbor N atom in the sensor. The added fluoride anion could capture the proton in the O-H moiety and the corresponding phenolic anion is formed, which could inhibit the ESPT process. The experimental UV/Vis and fluorescence spectra are well reproduced by the calculated vertical excitation energies. Frontier molecular orbital analysis indicates that the local excited state of phenolic anion is responsible for its enhanced fluorescence. Due to this reason, the sensor can be used to sense fluoride anion by monitoring the fluorescent change.


Optimization of an implantable collamer lens sizing method using high-frequency ultrasound biomicroscopy.
To develop and evaluate a new implantable collamer lens (ICL) sizing method that uses high-frequency ultrasound biomicroscopy (UBM). Interventional case series. ICL were implanted in 47 eyes of 25 patients (8 male, 17 female, mean age 34.7±6.8 years). At 3 months after ICL surgery, the vault (distance between ICL and crystalline lens) was measured using anterior optical coherence tomography (OCT). Stepwise multiple regression analysis was performed in order to determine the optimal ICL size that will be expected to achieve a 0.5-mm vault, and the regression equation was calculated. Mean keratometric power, axial length, anterior chamber depth, sulcus-to-sulcus (STS) diameter, and distance between STS plane and crystalline lens were applied as candidates for explanatory variables. Subsequently, the equation was applied to a new group of patients (81 eyes of 43 patients, 20 male and 23 female, mean age 35.6±7.2 years) in order to decide the ICL size. Postoperative vault was evaluated at 3 months after surgery. Main outcome measures were mean postoperative vault and percentages of eyes that achieved moderate vault. The regression equation was determined using 3 explanatory variables: anterior chamber depth (ACD), STS diameter, and distance between STS plane and anterior crystalline lens surface (STSL). The mean vault error (postoperative vault-predicted vault) was -0.06±0.29 mm. Of the total number of eyes, 88.9% had a vault between 0.15 and 1.0 mm. None of the eyes had a low vault (<0.15 mm). Nine eyes (11.1%) had a high vault (>1.0 mm). A novel ICL sizing equation was developed and shown to be an effective method for calculation of the optimal ICL size in order to achieve an appropriate vault.


Histone methyltransferase enzyme enhancer of zeste homolog 2 counteracts ischemic brain injury via H3K27me3-mediated regulation of PI3K/AKT/mTOR signaling pathway.
Epigenetic histone methylation plays a crucial role in cerebral ischemic injury, particularly in the context of ischemic stroke. However, the complete understanding of regulators involved in histone methylation, such as Enhancer of Zeste Homolog 2 (EZH2), along with their functional effects and underlying mechanisms, remains incomplete. Here, we employed a rat model of MCAO (Middle cerebral artery occlusion) and an OGD (Oxygen-Glucose Deprivation) model of primary cortical neurons to study the role of EZH2 and H3K27me3 in cerebral ischemia-reperfusion injury. The infarct volume was measured through TTC staining, while cell apoptosis was detected using TUNEL staining. The mRNA expression levels were quantified through quantitative real-time polymerase chain reaction (qPCR), whereas protein expressions were evaluated via western blotting and immunofluorescence experiments. The expression levels of EZH2 and H3K27me3 were upregulated in OGD; these expression levels were further enhanced by GSK-J4 but reduced by EPZ-6438 and AKT inhibitor (LY294002) under OGD conditions. Similar trends were observed for mTOR, AKT, and PI3K while contrasting results were noted for UTX and JMJD3. The phosphorylation levels of mTOR, AKT, and PI3K were activated by OGD, further stimulated by GSK-J4, but inhibited by EPZ-6438 and AKT inhibitor. Inhibition of EZH2 or AKT effectively counteracted OGD-/MCAO-induced cell apoptosis. Additionally, inhibition of EZH2 or AKT mitigated MCAO-induced infarct size and neurological deficit in vivo. Collectively, our results demonstrate that EZH2 inhibition exerts a protective effect against ischemic brain injury by modulating the H3K27me3/PI3K/AKT/mTOR signaling pathway. The results provide novel insights into potential therapeutic mechanisms for stroke treatment.


Stress, Burden, and Well-Being in Dementia and Nondementia Caregivers: Insights From the Caregiving Transitions Study.
Few population-based studies have directly compared caregivers of persons with dementia to caregivers of persons with other disabilities (nondementia caregivers). We enrolled dementia and nondementia caregivers who were providing substantial and sustained care and compared these groups on measures of caregiver stressors, appraisals of burden, and well-being. Caregivers (N = 251) who provided continuous care for at least 1 year and at least 5 h per week were recruited from the population-based REasons for Geographic And Racial Differences in Stroke (REGARDS) study. Caregivers reported on dementia caregiving status, stressors, burden, and well-being. Forty-seven percent (n = 117) reported caring for a person with dementia. Dementia caregivers reported more stressors, providing more care for self-care and behavioral problems than nondementia caregivers. Dementia caregivers also reported higher appraisals of stress and burden, and more depressive symptoms, but did not differ from nondementia caregivers on mental and physical health quality of life. In multivariable-adjusted models, adjustment for the total number of care recipient problems attenuated differences between dementia and nondementia caregivers on burden and depression measures. Dementia and nondementia caregivers showed relatively few differences in indicators of overall well-being in this population-based sample, perhaps because both groups of caregivers in this study were providing substantial care. Dementia caregivers may require special assistance with dementia-specific problems such as behavioral problems. Clinical interventions and policy changes targeting highly burdened caregivers are needed to support them in allowing their care recipients to age in place at home.


Structural studies of the antigenic polysaccharide of Eubacterium saburreum, strain T27.
The antigenic polysaccharide produced by Eubacterium saburreum, strain T27, is a homoglycan composed of D-glycero-D-galacto-heptose (Hep) residues having a nonasaccharide repeating-unit with the structure (leads to 6)-[alpha-Hepf-(1 leads to 4)]-beta-Hepp-(1) leads to (3)6)-[alpha-Hepf-(1 leads to 2), alpha-Hepf-(1 leads to 4)]-beta-Hepp-(1 leads to. The polysaccharide contains acetyl groups linked to O-2 (except to the 2,4,6-linked heptopyranosyl residue), O-3 and O-7 of part of both heptopyranosyl and heptofuranosyl residues. The assignment of an acetyl group at O-3 of part of the terminal heptofuranosyl and 4,6-linked heptopyranosyl groups is tentative.


Impact of hospital lockdown secondary to COVID-19 and past pandemics on surgical practice: A living rapid systematic review.
The COVID-19 pandemic has disrupted surgical practice worldwide. There is widespread concern for surgeon and provider safety, and the implications of hospital lockdown on patient care during epidemics. Medline, EMBASE, CENTRAL, and PubMed were systematically searched from database inception to July 1, 2020 and ongoing monthly surveillance will be conducted. We included studies that assessed postoperative patient outcomes or protection measures for surgical personnel during epidemics. We included 61 studies relevant to the COVID-19 pandemic and past epidemics. Lockdown measures were noted globally including cancellation of elective surgeries and outpatient clinics. The pooled postoperative complication rate during epidemics was 21.0% among 2095 surgeries. 31 studies followed the health of surgical workers with the majority noting no adverse outcomes with proper safety measures. This review highlights postoperative patient outcomes during worldwide epidemics including the COVID-19 pandemic and identifies specific safety measures to minimize infection of healthcare workers.


The dilemma of essential drugs in primary health care.
This article describes the evolution of essential drug programmes in eastern Africa from national donor assisted programmes to community-based drug programmes. Problems of the transition towards cost recovery are discussed and greater dialogue between government, donors and communities will be required if real community management is to be commenced.


VP4/VP56/VP35 Virus-like Particles Effectively Protect Grass Carp (Ctenopharyngodon idella) against GCRV-II Infection.
Grass carp reovirus (GCRV) seriously threatens the grass carp (Ctenopharyngodon idella) industry. Prophylactic GCRV vaccines prepared by virus-like particle (VLP) assembly biotechnology can improve effectiveness and safety. The highly immunogenic candidate antigens of GCRV vaccines that have been generally considered are the outer capsid proteins VP4, VP56, and VP35. In this study, VP4, VP56, and VP35 were expressed in an Escherichia coli expression system and a Pichia pastoris expression system. The successful assembly of uniform, stable, and non-toxic VP4/VP56/VP35 VLPs was confirmed through various assays. After vaccination and GCRV infection, the survival rate in the VLPs + adjuvant Astragalus polysaccharide (APS) group was the highest (62%), 40% higher than that in control group (22%). Through the antibody levels, tissue viral load, and antioxidant immunity assays, the P. pastoris VLP vaccine effectively improved IgM levels, alleviated tissue virus load, and regulated antioxidant immune-related indicators. The treatment with P. pastoris VLPs enhanced the mRNA expression of important immune-related genes in the head kidney, as measured by qRT-PCR assay. Upon hematoxylin-eosin staining examination, relatively reduced tissue pathological damage was observed in the VLPs + APS group. The novel vaccine using P. pastoris VLPs as an effective green biological agent provides a prospective strategy for the control of fish viral diseases.


Adult Cardiomyocyte Proliferation: a New Insight for Myocardial Infarction Therapy.
Myocardial infarction leads to cardiomyocyte loss, ensuing ventricular pathological remodeling, dramatic impairment of cardiac function, and ultimately heart failure. Unfortunately, the existing therapeutical treatments cannot directly replenish the lost myocytes in the injured myocardium and the long-term prognosis of heart failure after myocardial infarction remains poor. Growing investigations have demonstrated that the adult mammalian cardiomyocytes possess very limited proliferation capacity, and that was not enough to restore the injured heart. Recently, many studies were targeting to promote cardiomyocyte proliferation via inducing cardiomyocyte cell cycle re-entry for cardiac repair after myocardial infarction. Indeed, these results showed it is a feasible way to stimulate terminally differentiated cardiomyocyte proliferation. Here, we reviewed the major mechanisms and the potential targets for stimulating mammalian adult cardiomyocyte proliferation specifically. This will provide a new therapeutic strategy for the clinical treatment of myocardial infarction by activating the endogenous regeneration. Graphical abstract.


Prooxidant activity and toxicity of nordihydroguaiaretic acid in clone-9 rat hepatocyte cultures.
Nordihydroguaiaretic acid (NDGA) is a polyphenol. It is present at high concentrations in the leaves of the evergreen desert shrub, Larrea tridentate (Creosote bush), which has a long history of medicinal use traditionally by the native Americans and Mexicans. It is generally believed that the antioxidant properties of NDGA are responsible for the medicinal value of this desert shrub. The clone-9 rat hepatocyte cultures were used as an in vitro model to assess the hepatotoxic potential of NDGA and to determine whether it exhibits any prooxidant activity. The hepatocyte cultures were treated with NDGA for 2 h at 37 degrees C at concentrations of 0-100 microM. After the treatment period the cells, the culture supernatants and cell lysates were assayed for evaluation of prooxidant activity and toxicity of NDGA. Oxidative stress level and oxidative cell injury as measured by the peroxidation of membrane lipids and DNA double-strand breaks were used to index prooxidant activity. Cytotoxicity as measured by the leakage of the liver enzyme lactate dehydrogenase (LDH) into the culture medium, mitochondrial function and extent of cell proliferation were used as the endpoints of toxicity. Significant concentration-dependent differences were observed in these biomarkers over the concentration range examined demonstrating the prooxidant activity and toxicity of NDGA in clone-9 rat hepatocyte cultures.


Unveiling the immunomodulatory properties of Haemonchus contortus adhesion regulating molecule 1 interacting with goat T cells.
Gastrointestinal nematodes could release excretory-secretory (ES) proteins into the host environment to ensure their survival. These ES proteins act as immunomodulators to suppress or subvert the host immune response via the impairment of immune cell functions, especially in chronic infections. In our preliminary study, Haemonchus contortus adhesion-regulating molecule 1 (HcADRM1) was identified from H. contortus ES proteins (HcESPs) that interacted with host T cells via liquid chromatography-tandem mass spectrometry analysis. However, little is known about HcADRM1 as an ES protein which may play a pivotal role at the parasite-host interface. Based on bioinformatics approaches, multiple amino acid sequence alignment was conducted and the evolutionary relationship of HcADRM1 with ADRM1 orthologues was extrapolated. Employing RT-qPCR and immunohistochemistry assays, temporal transcriptional and spatial expression profiles of HcADRM1 were investigated. Using immunostaining approaches integrated with immunological bioassays, the immunomodulatory potentials of HcADRM1 on goat T cells were assessed. We hereby demonstrated that HcADRM1 with immunodiagnostic utility was a mammalian ADRM1 orthologue abundantly expressed at all developmental stages of H. contortus. Given the implications of ADRM1 proteins in cell growth, survival and development, we further investigated the immunomodulatory property of HcADRM1 as an individual ES protein acting at the parasite-host interface. The rHcADRM1 stimuli notably suppressed T cell viability, promoted intrinsic and extrinsic T cell apoptosis, inhibited T cell proliferation and induced cell cycle arrest at G1 phase. Simultaneously, rHcADRM1 stimuli exerted critical controls on T cell cytokine secretion profiles, predominantly by restraining the secretions of interleukin (IL)-4, IL-10 and interferon-gamma. Importantly, HcADRM1 protein may have prophylactic potential for anti-H. contortus vaccine development. Together, these findings may contribute to the clarification of molecular and immunomodulatory traits of ES proteins, as well as improvement of our understanding of parasite immune evasion mechanism in H. contortus-host biology.


[Analysis of motivations for antiflu vaccination of the Clermont-Ferrand University Hospital staff].
The vaccination of hospital staff decreases patient mortality and disorganization of services due to sick leave. The main aim of our study was to determine the Clermont-Ferrand University hospital (CHU) personnel's motivations for or against antiflu vaccination to increase the effectiveness of prevention campaigns. An autoquestionnaire with multiple choices was given to the 7601 CHU staff in May 2005. It documented socioprofessional characteristics, vaccinal status, and reasons for vaccination, nonvaccination, or stopping antiflu vaccination. The answer rate was 26.5% (2011 autoquestionnaires returned) and representative of the CHU staff. The rate of vaccination in 2004 was 36.35% with a strong correlation between vaccination and former influenza infection (p<0.001). Five of the six principal reasons for vaccination were altruistic including the first two: avoiding transmission to patients (61.8%) and his family (59.8%). The main reason for stopping was the lack of time. The reasons for nonvaccination were linked to a feeling of invulnerability: conviction of not being at risk, of being too young, or in good health. The Haute Autorité de santé's objective of vaccinal coverage against influenza for 75% of the health professionals requires their active compliance. The effectiveness of future campaigns could aim at improving knowledge by insisting particularly on the young age of the risk populations. The lack of time can be compensated by offering on-site vaccination, including at night and by proposing larger schedules for vaccination.


Systematic review and meta-analysis on the use of human platelet lysate for mesenchymal stem cell cultures: comparison with fetal bovine serum and considerations on the production protocol.
Mesenchymal stem cell (MSC) culturing for cell therapies needs a step forward to be routinely used in clinical settings. Main concerns regard the use of animal origin reagents, in particular supplementing the culture medium with FBS. Lately, Human Platelet Lysate (HPL) has been proposed as animal-free alternative, described as an excellent supplement for culturing MSCs. The aim of this systematic review was to analyze the current literature on the effect of HPL and FBS on ASCs and BMSCs. The primary outcome was the proliferation rate of cells cultured with FBS and HPL. Differences in terms of doubling time (DT) and population doubling (PD) were evaluated by meta-analysis, subgrouping data according to the cell type. A total of 35 articles were included. BMSCs and ASCs were used in 65.7% (23) and 28.6% (10) studies, respectively. Only two studies included both cell types. Overall, 22 studies were eligible for the meta-analysis. Among them, 9 articles described ASCs and 13 BMSCs. The results showed that BMSCs and ASCs cultured with 10% HPL and 5% HPL have lower DT and higher PD compared to cells cultured with 10% FBS. A possible correlation between the DT decrease and the application of at least 3 freeze/thaw cycles to induce platelet lysis was found. Additionally, HPL increased VEGF secretion and maintained the immuno-modulatory abilities for both cell types. The clarification reported here of the higher efficiency of HPL compared to FBS can help the transition of the scientific community towards clinical-related procedures. 1. The meta-analysis shows that HPL induces a population doubling increase and a doubling time decrease of both ASCs and BMSCs compared to FBS. 2. When at least 3 freeze/thaw cycles are applied to induce platelet lysis, the doubling time of HPL-cultured cells is lower than FBS-cultured cells (Created with BioRender.com).


Synthesis of novel trifluoromethyl substituted furo[2,3-b]pyridine and pyrido[3',2':4,5]furo[3,2-d]pyrimidine derivatives as potential anticancer agents.
A series of novel trifluoromethyl substituted furo[2,3-b]pyridine and pyrido[3',2':4,5]furo[3,2-d] pyrimidine derivatives 3a-b, 6a-k, 9, 10a-b, 11a-c and 12a-c were prepared from 2-carbethoxy-3-amino-6-trifluoromethyl furo[2,3-b]pyridine 1 under different set of conditions. Compounds functionalized with oxadiazole 11a-c were also prepared from 2-carbohydrazide-3-amino-6-trifluoromethyl furo[2,3-b]pyridine 4. All the final products were screened for anticancer activity against four human cancer cell lines such as Neuro-2a, Hela, A549 and COLO 205 as well as normal human lung cell line, IMR-90. All the compounds showed promising anticancer activity against all the tested cell lines at <25 μM concentration except 5b, 6d, 6e and 6k. The selectivity index (SI) values have also been calculated for all the tested compounds in comparison to the normal cell line. Compounds 6g, 10a, 10b, and 11a were considered as potential leads which showed cytotoxicity with IC50 values of 10, 10.7, 11.0 and 10.5 μM, respectively. Compounds 7 and 12a were considered as highly potent exhibiting promising cytotoxicity with IC50 value of 5.8 and 3.6 μM, respectively.


Synthesis and pharmacological evaluation of carbamic acid 1-phenyl-3-(4-phenyl-piperazine-1-yl)-propyl ester derivatives as new analgesic agents.
A series of carbamic acid 1-phenyl-3-(4-phenyl-piperazine-1-yl)-propyl ester derivatives were synthesized through discovery strategies for balancing target-based in vitro screening and phenotypic in vivo screening. All the newly synthesized compounds were screened for their analgesic activities and compared with standard drug morphine. Among them, compound 44r, a potent analgesic agent that has favorable pharmacokinetic properties in rats and most importantly, has a wide safety margin. We demonstrated with in vitro and in vivo functional assays that its analgesic activity might be through 5-HT(2A) antagonism to some extent. Hence, it is concluded that there is ample scope for further study in developing compound 44r as a good lead candidate for an analgesic agent.


A DNAzyme based knockdown model for Fragile-X syndrome in zebrafish reveals a critical window for therapeutic intervention.
FXS is the leading cause of intellectual disabilities in males and a major monogenic cause of ASD (Autism spectrum disorders). It occurs due to the loss of FMRP, whose role in early development is not well understood. In this study, we have used a novel DNAzyme based approach to create a larval model of FXS in zebrafish with specific focus on the early developmental window. Fmr1specific DNAzymes were electroporated into embryos to create the knockdown. Changes in RNA and protein levels of FMRP and relevant biomarkers were measured in the 0-7dpf window. Behavioral tests to measure anxiety, cognitive impairments and irritability in the larvae were conducted at the 7dpf stage. Drug treatment was carried out at various time points in the 0-7dpf window to identify the critical window for pharmacological intervention. The DNAzyme based knockdown approach led to a significant knockdown of FMRP in the zebrafish embryos, accompanied by increased anxiety, irritability and cognitive impairments at 7dpf, thus creating a robust larval model of FXS. Treatment with the Mavoglurant was able to rescue the behavioral phenotypes in the FXS larvae, and found to be more efficacious in the 0-3dpf window. The results from this study have revealed that a) a DNAzyme based knockdown approach can be used to create robust larval zebrafish model of disease, in a high-throughput manner and b) optimal window for therapeutic intervention for FXS as well as other pediatric diseases with a monogenic cause can be identified using such a model.


Determinants of severity among hospitalised COVID-19 patients: Hospital-based case-control study, India, 2020.
Risk factors for the development of severe COVID-19 disease and death have been widely reported across several studies. Knowledge about the determinants of severe disease and mortality in the Indian context can guide early clinical management. We conducted a hospital-based case control study across nine sites in India to identify the determinants of severe and critical COVID-19 disease. We identified age above 60 years, duration before admission >5 days, chronic kidney disease, leucocytosis, prothrombin time > 14 sec, serum ferritin >250 ng/mL, d-dimer >0.5 ng/mL, pro-calcitonin >0.15 μg/L, fibrin degradation products >5 μg/mL, C-reactive protein >5 mg/L, lactate dehydrogenase >150 U/L, interleukin-6 >25 pg/mL, NLR ≥3, and deranged liver function, renal function and serum electrolytes as significant factors associated with severe COVID-19 disease. We have identified a set of parameters that can help in characterising severe COVID-19 cases in India. These parameters are part of routinely available investigations within Indian hospital settings, both public and private. Study findings have the potential to inform clinical management protocols and identify patients at high risk of severe outcomes at an early stage.


The signaling network of transforming growth factor beta1, protein kinase Cdelta, and integrin underlies the spreading and invasiveness of gastric carcinoma cells.
Integrin-mediated cell adhesion and spreading enables cells to respond to extracellular stimuli for cellular functions. Using a gastric carcinoma cell line that is usually round in adhesion, we explored the mechanisms underlying the cell spreading process, separate from adhesion, and the biological consequences of the process. The cells exhibited spreading behavior through the collaboration of integrin-extracellular matrix interaction with a Smad-mediated transforming growth factor beta1 (TGFbeta1) pathway that is mediated by protein kinase Cdelta (PKCdelta). TGFbeta1 treatment of the cells replated on extracellular matrix caused the expression and phosphorylation of PKCdelta, which is required for expression and activation of integrins. Increased expression of integrins alpha2 and alpha3 correlated with the spreading, functioning in activation of focal adhesion molecules. Smad3, but not Smad2, overexpression enhanced the TGFbeta1 effects. Furthermore, TGFbeta1 treatment and PKCdelta activity were required for increased motility on fibronectin and invasion through matrigel, indicating their correlation with the spreading behavior. Altogether, this study clearly evidenced that the signaling network, involving the Smad-dependent TGFbeta pathway, PKCdelta expression and phosphorylation, and integrin expression and activation, regulates cell spreading, motility, and invasion of the SNU16mAd gastric carcinoma cell variant.


Role of 3D intraoperative imaging in orthopedic and trauma surgery.
Intraoperative three-dimensional (3D) imaging is now feasible because of recent technological advances such as 3D cone-beam CT (CBCT) and flat-panel X-ray detectors (FPDs). These technologies reduce the radiation dose to the patient and surgical team. The aim of this study is to review the advantages of 3D intraoperative imaging in orthopedic and trauma surgery by answering the following 5 questions: What are its technical principles? CBCT with a FPD produces non-distorted digital images and frees up the surgical field. The high quality of these 3D intraoperative images allows them to be integrated into surgical navigation systems. Human-robot comanipulation will likely follow soon after. Conventional multislice CT technology has also improved to the point where it can be used in the operating room. What can we expect from 3D intraoperative imaging and which applications have been validated clinically? We reviewed the literature on this topic for the past 10 years. The expected benefits were determined during the implantation of pedicular screws: more accurate implantation, fewer surgical revisions and time savings. There are few studies in trauma or arthroplasty cases, as robotic comanipulation is a more recent development. What is the tolerance for irradiation to the patient and surgical team? The health drawbacks are the harmful radiation-induced effects. The deterministic effects that we will develop are correlated to the absorbed dose in Gray units (Gy). The stochastic and carcinogenic effects are related to the effective dose in milliSievert (mSv) of linear evolution without threshold. The International Commission on Radiological Protection (ICRP) states that irradiation for medical purposes with risk of detriment is acceptable if it is justified by an optimization attempt. The radioprotection limits must be known but do not constitute opposable restrictions. The superiority of intraoperative 3D imaging over fluoroscopy has been demonstrated for spine surgery and sacroiliac screw fixation. How does the environment need to be adapted? The volume, access, wall protection and floor strength of the operating room must take into account the features of each machine. The instrumentation implants and need for specialized staff result in additional costs. Not every system can track movements during the CBCT acquisition thus transient suspension of assisted ventilation may be required. Is it financially viable? This needs to be calculated based on the expected clinical benefits, which mainly correspond to the elimination of expenses tied to surgical revisions. Our society's search for safety has driven the investments in this technology. LEVEL OF EVIDENCE: V, Expert opinion.


Safety and feasibility of three-dimensional McKeown minimally invasive esophagectomy.
To compare the perioperative outcomes from McKeown minimally invasive esophagectomy (MIE) when performed in three-dimensional versus two-dimensional visualization system, and investigate the learning curve of a single surgeon who implemented three-dimensional McKeown MIE. A total of 335 consecutive cases (three-dimensional or two-dimensional) were identified. Perioperative clinical parameters were compared and cumulative sum learning curve was plotted. Propensity score matching was used to reduce selection bias from confounding factors. Patients in three-dimensional group were associated with more chronic obstructive pulmonary disease (23.9% vs 3.0%, p < 0.01). After propensity score matching (108 matched patients in each groups), this finding was no longer statistically significant. Comparing to two-dimensional group, significant improvement in total retrieved lymph nodes (28 vs 33, p = 0.003) was observed in three-dimensional group. In addition, more lymph nodes around the right recurrent laryngeal nerve were harvested in three-dimensional group than that in two-dimensional group (p = 0.045). However, there were no significantly differences were found between the two groups in terms of other intraoperative parameters (e.g., operative time) and postoperative relevant outcomes (e.g., lung infection). Furthermore, the change point in the cumulative sum learning curves for intraoperative blood loss and thoracic procedure time was 33 procedures, respectively. Three-dimensional visualization system appears to be superior in performing lymphadenectomy during McKeown MIE to that of a two-dimensional technique. For surgeons proficient in performing two-dimensional McKeown MIE, the learning curve for a three-dimensional procedure appears to begin near proficiency after more than 33 cases.


Associations between nutrients in one-carbon metabolism and fetal DNA methylation in pregnancies with or without gestational diabetes mellitus.
Gestational diabetes mellitus (GDM), characterized by hyperglycemia that develops during pregnancy, increases the risk of fetal macrosomia, childhood obesity and cardiometabolic disorders later in life. This process has been attributed partly to DNA methylation modifications in growth and stress-related pathways. Nutrients involved with one-carbon metabolism (OCM), such as folate, choline, betaine, and vitamin B12, provide methyl groups for DNA methylation of these pathways. Therefore, this study aimed to determine whether maternal OCM nutrient intakes and levels modified fetal DNA methylation and in turn altered fetal growth patterns in pregnancies with and without GDM. In this prospective study at a single academic institution from September 2016 to June 2019, we recruited 76 pregnant women with and without GDM at 25-33 weeks gestational age and assessed their OCM nutrient intake by diet recalls and measured maternal blood OCM nutrient levels. We also collected placenta and cord blood samples at delivery to examine fetal tissue DNA methylation of the genes that modify fetal growth and stress response such as insulin-like growth factor 2 (IGF2) and corticotropin-releasing hormone (CRH). We analyzed the association between maternal OCM nutrients and fetal DNA methylation using a generalized linear mixed model. Our results demonstrated that maternal choline intake was positively correlated with cord blood CRH methylation levels in both GDM and non-GDM pregnancies (r = 0.13, p = 0.007). Further, the downstream stress hormone cortisol regulated by CRH was inversely associated with maternal choline intake (r = - 0.36, p = 0.021). Higher maternal betaine intake and serum folate levels were associated with lower cord blood and placental IGF2 DNA methylation (r = - 0.13, p = 0.049 and r = - 0.065, p = 0.034, respectively) in both GDM and non-GDM pregnancies. Further, there was an inverse association between maternal betaine intake and birthweight of infants (r = - 0.28, p = 0.015). In conclusion, we observed a complex interrelationship between maternal OCM nutrients and fetal DNA methylation levels regardless of GDM status, which may, epigenetically, program molecular pathways related to fetal growth and stress response.


Mendelian randomization to evaluate the effect of plasma vitamin C levels on the risk of Alzheimer's disease.
Until now, observational studies have explored the impact of vitamin C intake on Alzheimer's disease (AD) risk, however, reported ambiguous findings. To develop effective therapies or prevention, the causal link between vitamin C levels and AD should be established. Here, we selected 11 plasma vitamin C genetic variants from a large-scale plasma vitamin C GWAS dataset (N = 52,018) as the potential instrumental variables. We extracted their corresponding summary statistics from large-scale IGAP clinically diagnosed AD GWAS dataset (N = 63,926) and UK Biobank AD proxy phenotype GWAS dataset (N = 314,278), as well as two UK Biobank subgroups including the maternal AD group (27,696 cases of maternal AD and 260,980 controls) and paternal AD group (14,338 cases of paternal AD and 245,941 controls). We then performed a Mendelian randomization (MR) study to evaluate the causal association between plasma vitamin C levels and the risk of AD and AD proxy phenotype. Meanwhile, we further verified these findings using a large-scale cognitive performance GWAS dataset (N = 257,841). In IGAP, we found no significant causal association between plasma vitamin C levels and the risk of AD. In UK Biobank, we found that per 1 SD increase in plasma vitamin C levels (about 20.2 μmol/l) was significantly associated with the reduced risk of AD proxy phenotype (OR = 0.93, 95% CI 0.88-0.98, P = 7.00E-03). A subgroup MR analysis in UK Biobank indicated that per 1 SD increase in plasma vitamin C levels could significantly reduce the risk of AD proxy phenotype in the maternal AD group (OR = 0.89, 95% CI 0.84-0.94, P = 7.29E-05), but not in the paternal AD group (OR = 1.02, 95% CI 0.92-1.12, P = 7.59E-01). The leave-one-out permutation further showed that the SLC23A1 rs33972313 variant largely changed the precision of the overall MR estimates in all these four GWAS datasets. Meanwhile, we did not observe any significant causal effect of plasma vitamin C levels on the cognitive performance. We demonstrated that there may be no causal association between plasma vitamin C levels and the risk of AD in people of European descent. The insistent findings in clinically diagnosed AD and AD proxy phenotype may be caused by the phenotypic heterogeneity.


Regeneration of the donor side after autogenous fibula transplantation in 53 patients: evaluation by dual x-ray absorptiometry.
We determined the degree of fibular regeneration at the donor site, using radiographs and dual x-ray absorptiometry, in 53 patients who underwent autogenous nonvascularized fibular transplantation for tumor reconstruction in long bones (mean follow-up 15 (3-26) years). Logistic regression was used to determine whether gender, age at transplantation, time since transplantation, bone mineral density (BMD), and length of the graft were associated with fibular regeneration. 26 patients had spontaneous complete bone regeneration. Younger age at transplant was the only predictor of fibular regeneration. In predicting fibular regeneration, sensitivity was 96% and specificity 74%, using 15 years of age as a cut-off. In the long-term follow-up, we found only gradual changes in the BMD and the values ranged from 24% to 217%. We found no correlations of bone mineral density with age, gender, length of the graft, or time since transplantation.


Mutations in the C-terminus of the X protein of hepatitis B virus regulate Wnt-5a expression in hepatoma Huh7 cells: cDNA microarray and proteomic analyses.
The hepatitis B virus x gene (HBx) is a promiscuous transactivator implicated in the development of hepatocellular carcinoma (HCC). The present study was designed to investigate the molecular events regulated by HBx. Genomic and proteomic expression profiling was performed in Huh7 HCC cells transfected with HBx mutants with a C-terminal deletion. The gene and protein expression of wingless-type murine-mammary-tumour virus (MMTV) integration site family, member 5A (Wnt-5a) was validated by analyses of reverse transcription-polymerase chain reaction (RT-PCR), real-time RT-PCR, western blot and immunohistochemistry. Differentially expressed genes and proteins were found in the transfected Huh7 HCC cells; most of them were involved in transcriptional regulation, although others including oncogenes or tumor suppressor genes, and molecules involved in cell junctions, signal transduction pathways, metabolism or the immune response were also observed. The expression of the Wnt-5a gene was elevated >10-fold in Huh7 cells transfected with the HBx3'-30 amino acid deletion mutant. However, the expression was downregulated by the transfection with the HBx3'-40 amino acid deletion mutant. The changes in Wnt-5a expression were also observed in human HCC tissues, compared with corresponding non-cancerous liver tissues. A negative correlation was found between the expression of Wnt-5a and HBx COOH mutations in HCC tissues. HBx mutants may participate in the development and progression of HCC, at least in part through the Wnt-5a pathway.


Mitochondrial metabolism of guanine nucleotides. Possible role of guanosine.
The catabolism of intramitochondrial guanine nucleotides was examined. During 30 min incubation of rat liver mitochondria at 37 degrees C in the presence of oligomycin and carboxyatractyloside, guanine and xanthine were formed and appeared in the medium. Under these conditions, the direct conversion of GMP to guanine by hypoxanthine-guanine phosphoribosyltransferase is suggested to be the main catabolic route within the organelles. Only very small amounts of guanosine were produced and detected both inside and outside the organelles. [14C]Guanosine and [14C]inosine were taken up by the mitochondria. Therefore, guanosine is suggested to be a precursor of intramitochondrial guanine nucleotides.


[Identification and assessment of multiple human papillomavirus types in condyloma acuminata lesions from patients with genital warts in Beijing area].
To identify and assess multiple human papillomavirus types in condyloma acuminatum lesions from patients with genital warts in Beijing area, and compare different features between otherwise healthy and immunosuppressed patients. PCR, RFLP and nucleotide sequencing analysis were used to determine HPV types from individual lesions. The predominant type from other healthy patients was HPV6, secondly HPV11. The mean age of patients infected by HPV6 was lower than that of HPV11 and HPV6 + 11. While lesions from immunosuppressed patients were often contained HPV11 or mixed with HPV6. Besides, HPV types 16 and 53 were detected from infected lesions than other HPV types. HPV6 was the major pathogen of condyloma acuminatum, but infected patients were at lower ages. While HPV11 was most often detected from immunosuppressed patients. As a low risk virus in normal genital tract, HPV53 also could be a pathogen in genital warts.


Efficient preparation of incensole and incensole acetate, and quantification of these bioactive diterpenes in Boswellia papyrifera by a RP-DAD-HPLC method.
Incensole and incensole acetate, found in incense, are encouraging potent bioactive diterpenic cembrenoids, inhibiting Nuclear Factor-kappaB activation. Furthermore, incensole acetate elicits psycho-activity in mice by activating the TRPV3 channels in the brain. Starting from crude extracts of the incense species Boswellia papyrifera Hochst., a convenient procedure for the efficient large-scale synthesis of incensole and its acetate is presented. Additionally, a reversed-phase, diode-array-detection, high-performance liquid chromatography (RP-DAD-HPLC) method for the quantification of incensole and incensole acetate is reported, indicating that these two compounds are typical biomarkers for B. papyrifera.


Surgical treatment of lung cancer in patients over the age of 70 years.
Eighty patients of 70 years of age with lung cancer have been treated since 1964. Forty-eight received no therapy or chemotherapy and/or irradiation. Mean survival was 3.5 to 10 months, and only three (6%) survived for 2 years. Twenty-two of 32 patients selected for thoracotomy underwent resection for cure (70%). The operative mortality rate was 18% for resection, and 64% of the patients survived for at least 2 years. The incidence of exploratory thoracotomy decreased in the last 5 years of the study with the introduction of more rigorous preoperative evaluation. The poor survival rate without resection and the favorable survival rate following recovery from resection support the continued use of surgical resection for lung cancer in elderly patients, despite an increased operative risk.


Anaesthetic management of a case of nesidioblastosis for subtotal pancreatectomy.
Hyperinsulinism is a rare cause of severe persistent hypoglycaemia in the neonatal period. It is associated with a high incidence of brain damage and mental retardation as a consequence of repeated episodes of hypoglycaemia. Subtotal to near total pancreatectomy is indicated as a matter of urgency to decrease the amount of circulating insulin. The perioperative management of a 45-day-old, 5 kg male infant with hyperinsulinaemia (nesidioblastosis) is described. He had a history of generalized tonic clonic seizures 4 h after birth. The blood sugar at that time was 0.66 mmol x l(-1) (12 mg x dl(-1)) and serum calcium was 2.4 mmol x l(-1) (9.82 mg x dl(-1)). The insulin : glucose ratio was 1.6 (normal < 0.4). Occasional episodes of hypoglycaemia persisted in spite of medical line of management with intravenous dextrose 12%, 2 h gastric tube feeds, hydrocortisone (5 mg x kg(-1) x day(-1) i.v.) and oral diazoxide 10 mg x kg(-1), 8 h for 3 weeks. A CT scan and USG did not reveal any abnormality of the pancreas. However, the EEG varied from one of abnormally low amplitude to an isoelectric record. Renal, liver function tests and coagulation profile were normal. The patient was scheduled for elective subtotal pancreatectomy. The anaesthetic management with emphasis on glucose homeostasis and fluid balance is discussed.


Activity of the Keggin polyoxotungstate PM-19 against herpes simplex virus type 2 infection in immunosuppressed mice: role of peritoneal macrophage activation.
The in vivo antiviral activity of the Keggin polyoxotungstate PM-19 [K7(PTi2W10O40).6H2O] against herpes simplex virus type 2 (HSV-2) was investigated in mice immunosuppressed by cyclophosphamide (CY). When PM-19 was administered intraperitoneally to immunosuppressed mice for 3 days (once daily) starting at the time of infection, it prevented death due to HSV-2 encephalitis in a dose-dependent manner (10-25 mg/kg). The in vivo anti-HSV-2 activity of PM-19 was superior to that of acyclovir. Intraperitoneal administration of PM-19 to the immunosuppressed mice significantly increased the number of peritoneal cells, especially macrophages. PM-19 did not stimulate interferon-inducing activity or natural killer cell activity, but markedly enhanced peritoneal macrophage functions: (1) phagocytic activity as assessed by measuring the amount of 51Cr-labeled sheep red blood cells taken into the macrophages, and (2) extrinsic antiviral activity as monitored by reduction in the numbers of plaque formed upon cocultivation of HSV-2-infected HEL cells with the macrophages. These results point to the role of peritoneal macrophage activation in the activity of PM-19 against HSV-2 infection in immunosuppressed mice.


Micro-RNA expression profile of BALB/c mouse glandular stomach in the early phase of Cryptosporidium muris infection.
Cryptosporidiosis is a zoonotic disease in humans and animals that is caused by infection with the oocysts of Cryptosporidium. MicroRNAs (miRNAs) are important players in regulating the innate immune response against parasitic infection. Public miRNAs data for studying pathogenic mechanisms of cryptosporidiosis, particularly in natural hosts, are scarce. Here, we compared miRNA profiles of the glandular stomach of C. muris-infected and uninfected BALB/c mice using microarray sequencing. A total of 10 miRNAs (including 3 upregulated and 7 downregulated miRNAs) with significant differential expression (|FC| ≥ 2 and P value < 0.05) were identified in the glandular stomach of BALB/c mice 8 h after infection with C. muris. MiRWalk and miRDB online bioinformatics tools were used to predict the target genes of differentially expressed miRNAs. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed to annotate the target genes. GO analysis indicate that gene transcription-related and ion transport-related GO terms were significantly enriched. In addition, the KEGG analyses showed that the target genes were strongly related to diverse types of tumor disease progression and anti-pathogen immunity pathways. In the current study, we firstly report changes in miRNA expression profiles in the glandular stomach of BALB/c mice at the early phase of C. muris invasion. This dysregulation in miRNA expression may contribute to our understanding of cryptosporidiosis pathology. This study provides a new perspective on the miRNA regulatory mechanisms of cryptosporidiosis, which may help in the development of effective control strategies against this pathogen.


Effects of losartan or atenolol in hypertensive patients without clinically evident vascular disease: a substudy of the LIFE randomized trial.
Cardiovascular morbidity and mortality are reduced by treatment with the angiotensin II AT(1)-receptor antagonist losartan compared with conventional treatment with the beta-blocker atenolol in patients with hypertension and electrocardiogram-defined left ventricular hypertrophy, many of whom had known vascular disease. To determine whether losartan reduces cardiovascular event rates in lower-risk hypertensive patients without clinically evident vascular disease. Subgroup analysis of a randomized trial. The Losartan Intervention for Endpoint reduction in hypertension (LIFE) study. 6886 men and women (57% women) 55 to 80 years of age (average, 66 years) with essential hypertension (sitting blood pressure, 160 to 200/95 to 115 mm Hg [average, 174/98 mm Hg]) and electrocardiogram-defined left ventricular hypertrophy who did not have clinically evident vascular disease. Patients were randomly assigned to once-daily double-blind treatment with losartan or atenolol. An end point committee ascertained end points (cardiovascular death, stroke, or myocardial infarction). Blood pressure was reduced similarly by losartan and atenolol. The primary composite end point occurred in 282 losartan-treated patients (17.5 per 1000 patient-years) and 355 atenolol-treated patients (21.8 per 1000 patient-years; relative risk, 0.81 [95% CI, 0.69 to 0.95]; P = 0.008). Cardiovascular death occurred in 103 losartan-treated patients and 132 atenolol-treated patients (relative risk, 0.80 [CI, 0.62 to 1.04]; P = 0.092), stroke (nonfatal and fatal) occurred in 125 losartan-treated patients and 193 atenolol-treated patients (relative risk, 0.66 [CI, 0.53 to 0.82]; P < 0.001), and myocardial infarction (nonfatal and fatal) occurred in 110 losartan-treated patients and 100 atenolol-treated patients (relative risk, 1.14 [CI, 0.87 to 1.49]; P > 0.2). New-onset diabetes occurred less often in patients treated with losartan (n = 173) than in patients treated with atenolol (n = 254) (relative risk, 0.69 [CI, 0.57 to 0.84]; P < 0.001). Benefits of losartan treatment were numerically smaller, but not significantly so, in patients with preexisting vascular disease. In hypertensive patients without clinically evident vascular disease, losartan was more effective than atenolol in preventing cardiovascular morbidity and death, predominantly stroke, independent of blood pressure reduction.


A multidimensional impedance platform for the real-time analysis of single and combination drug pharmacology in patient-derived viable melanoma models.
In today's development of anticancer drugs, there is an enormous demand for sensitive, non-invasive real-time screening technologies to identify pharmacodynamics/-kinetics of single and combined drugs with high precision. The combination of sophisticated drug sensitivity testing with advanced in vitro tumor models reflecting heterogeneous tumor behavior in vivo is needed to more reasonably predict therapeutic outcome in vivo. In this study, the benefits of our real-time, non-invasive multidimensional impedance platform over standard in vitro drug sensitivity assays were demonstrated quantitatively using an advanced melanoma model. Detailed pharmacological profiles of clinically established targeted therapeutics in single and combination treatment have been identified in patient tissue and isolated 2D/3D cell line cultures. Impedance spectroscopy revealed significant differences in tissue structure responsible for BRAF inhibitor pharmacokinetics in BRAFV600E tumor microfragments and cell lines. Remarkably, BRAF-/MEK inhibitor combination treatment of direct patient-derived tissue, but not melanoma cell lines, resulted in short-term antagonistic effects consistent with in vivo findings. In contrast, the clinically validated resistance delay and thus long-term synergy of targeted therapeutics in advanced melanoma models has been demonstrated using impedance technology. The results demonstrate limited clinical transferability of 2D/3D cancer cell line-based chemosensitivity data and underline the importance of in vivo-like direct patient-derived tissue for predictive drug studies. Our non-invasive and highly sensitive multidimensional impedance platform offers great potential for quantifying short- and long-term drug kinetics and synergies to identify the most effective drug combinations in advanced cancer models, thereby improving personalized drug development and treatment planning and ultimately, overall patient outcomes.


Incidence of donor-specific antibodies in kidney transplant patients following conversion to an everolimus-based calcineurin inhibitor-free regimen.
Scarce data exist regarding the incidence of donor-specific antibodies (DSAs) in kidney transplant patients receiving everolimus-based immunosuppression without calcineurin inhibitors (CNIs). The aim of this retrospective case-control study was to compare the incidence of de novo DSAs in patients converted to an everolimus-based regimen without CNIs with that seen in patients maintained on CNIs. Sixty-one DSA-free kidney transplant patients who had been converted to an everolimus-based regimen (everolimus group) were compared to 61 other patients maintained on CNIs-based regimen (control group). Patients were matched according to age, gender, induction therapy, date of transplantation, and being DSA-free at baseline. At last follow-up, the incidence of DSAs was 9.8% in the everolimus group and 5% in the control group (p = ns). In the everolimus group, the increased incidence of DSAs between baseline and last follow-up was statistically significant. Antibody-mediated rejection occurred in 6.5% in the everolimus group and 0% in the CNIs group. The incidence of DSAs is numerically increased in kidney transplant patients treated with an everolimus-based without CNIs. A study including a larger number of patients is required to determine whether a CNI-free everolimus-based immunosuppression significantly increases DSAs formation.


Differential impact of admission C-reactive protein levels on 28-day mortality risk in patients with ST-elevation versus non-ST-elevation myocardial infarction (from the Monitoring Trends and Determinants on Cardiovascular Diseases [MONICA]/Cooperative Health Research in the Region of Augsburg [KORA] Augsburg Myocardial Infarction Registry).
The present study investigated the association between C-reactive protein (CRP) on admission independently and in combination with troponin and short-term prognosis in an unselected sample of patients with acute myocardial infarction (AMI) from the community. The study population consisted of 1,646 patients aged 25 to 74 years who were consecutively hospitalized with AMI within 12 hours after symptom onset. They were divided into the 2 groups of CRP positive (n = 919) or CRP negative (n = 727) with respect to admission CRP (cutoff < or =0.3 mg/dl). CRP-positive patients had significantly more in-hospital complications and a higher 28-day case-fatality rate (9.6% vs 3.4%; p <0.0001). Troponin at admission (n = 1,419) also correlated with 28-day case-fatality rate (troponin-negative 3.4% vs troponin-positive patients 8.0%; p <0.002). Multivariable analysis showed that both troponin positivity and CRP positivity were associated with a 2-fold (adjusted odds ratio 1.99, 95% confidence interval 1.15 to 3.44; adjusted odds ratio 2.05, 95% confidence interval 1.09 to 3.84, respectively) increased risk of dying within 28 days after the acute event for all patients with AMI. Stratifying by AMI type showed that in patients with ST-elevation myocardial infarction (STEMI), troponin positivity, but not CRP positivity, independently predicted 28-day case fatality. In patients with non-STEMI, CRP positivity, but not troponin positivity, predicted outcome. In conclusion, admission CRP was a powerful parameter for risk stratification of patients with AMI. Stratification by AMI type and troponin showed that CRP was a better short-term risk predictor for patients with non-STEMI, and troponin was, for patients with STEMI.


Structure-based functional annotation of hypothetical proteins from Candida dubliniensis: a quest for potential drug targets.
Candida dubliniensis is an emerging pathogenic yeast in humans and infections are usually restricted to mucosal parts of the body. However, its presence in specimens of immunocompromised individuals, especially in HIV-positive patients, is of major medical concern. There is a large fraction of genomes of C. dubliniensis in the database which are uncharacterized for their biochemical, biophysical, and/or cellular functions, and are identified as hypothetical proteins (HPs). Function annotation of Candida genome is, therefore, essentially required to facilitate the understanding of mechanisms of pathogenesis and biochemical pathways important for selecting novel therapeutic target. Here, we carried out an extensive analysis to explain the functional properties of genome, using available protein structure and function analysis tools. We successfully modeled the structures of eight HPs for which a template with moderate sequence similarity was available in the protein data bank. All modeled structures were analyzed and we found that these proteins may act as transporter, kinase, transferase, ketosteroid, isomerase, hydrolase, oxidoreductase, and binding targets for DNA and RNA. Since these unique HPs of Candida showed no homologs in humans, these proteins are expected to be a potential target for future antifungal therapy.


Identification of rat urinary glycoproteome captured by three lectins using gel and LC-based proteomics.
Many different types of urine proteome studies have been done, but urine glycoprotein studies are insufficient. Therefore, we studied the glycoproteins from rat urine, which could be used to identify biomarkers in an animal model. First, urinary proteins were prepared by using the dialysis and lyophilizing methods from rat urine. Glycoproteins enriched with lectin affinity purification, concanavalin A, jacalin and wheat germ agglutinin from the urinary proteins were separated by means of reverse-phase fast protein LC (FPLC) or 1-D PAGE. Each FPLC fraction and 1-D PAGE gel band were trypsin-digested and analyzed by means of nanoLC-MS/MS. LC-MS/MS analyses were carried out by using linear ion trap MS. A total of 318 rat urinary glycoproteins were identified from the FPLC fractions and gel bands; approximately 90% of identified proteins were confirmed as glycoproteins in Swiss-Prot. Many glycoproteins, known as biomarkers, including C-reactive protein, uromodulin, amyloid beta A4 protein, alpha-1-inhibitor 3, vitamin D-binding protein, kallikrein 3 and fetuin-A were identified in this study. By studying urinary glycoproteins collected from rat, these results may help to assist in identifying urinary biomarkers regarding various types of disease models.


Liver disease associated with ZZ alpha1-antitrypsin deficiency and ursodeoxycholic acid therapy in children.
To investigate the effect of ursodeoxycholic acid (UDCA) in children with liver disease associated with ZZ alpha1-antitrypsin (AAT) deficiency. A total of 42 affected children received UDCA (30 mg x kg x day(-1)) and underwent clinical and biochemical follow-up at least yearly. In group 1, 22 children whose mean initial gamma-glutamyl-transpeptidase (GGT) was 7.4 x N normalized serum liver test results after a mean treatment of 2.6 years. In 16 of these children, UDCA was discontinued. Relapse was observed in 11 children, and liver test results returned to normal after UDCA resumption. In the other 5 children, liver test results remained normal during a mean period of 2.5 years. In group 2, 11 children (mean initial GGT 12.8 x N) had improved liver test results after a mean treatment of 2.3 years. In group 3, 9 children (mean initial GGT 33.8 x N) had no liver test result improvement and evolution toward cirrhosis, requiring liver transplantation in 7. Most of the children in group 1 had normal results of clinical examination after UDCA treatment, versus none in group 3 (P < or = 0.00001). Initial GGT (P < or = 0.002) and total bilirubin (P < or = 0.05) levels were significantly lower in group 1 than in group 3. Combined initial values of GGT < or =5.5 x N and total bilirubin < or =66 micromol/L were associated with normalization of liver test results in 90% of children. UDCA may significantly improve clinical status and liver test results in some children with liver disease associated with ZZ AAT deficiency. No beneficial effect of UDCA was shown in children with the most severe liver involvement. Initial levels of GGT and total bilirubin may be of prognostic value for therapy effectiveness.


Evaluation of a subcutaneously implanted chamber for antibody production in rabbits.
Polyclonal antibody production in subcutaneous chambers was compared to traditional antibody production methods in rabbits. The chamber, a sterilized plastic wiffle golf ball that had been surgically implanted in the subcutis of the thoracic region, was immunized via a percutaneous injection of antigen into the core of the ball through one of the perforations in the chamber wall. Rabbits bearing chambers were immunized on the same schedule and with the same concentrations of antigens as were provided the adjuvant injected rabbits. Fluid volumes of 12 to 22 ml could be removed from each chamber at weekly intervals. Chamber antibody to specific microbial antigens was equal to or better than serum antibody produced to the same antigens with Freund's or acrylamide adjuvants. The comfort of the rabbit, the ease in chamber immunization, and the recovery of high titer antibody in large volumes make the subcutaneous chamber an attractive method for polyclonal antibody production.


Identification of mammalian GFRalpha-2 splice isoforms.
Neurturin (NTN) belongs to a structurally related family of bioactive molecules which include glial cell-line derived neutrotrophic factor (GDNF) and perserphin (PSP). NTN exerts its effects through a multicomponent receptor system which include a receptor (GFRalpha-2) and the proto-oncogene c-RET. We report here the identification of three splice isoforms of the GFRalpha-2 receptors (GFRalpha-2a, GFRalpha-2b and GFRalpha-2c) by reverse transcription-PCR (RT-PCR). GFRalpha-2b is a novel splice variant. All three isoforms were found to be expressed in various adult murine tissues as well as in the brain of the newborn human. The identity of these isoforms were further confirmed by the isolation of the gene and the characterisation of the splice junctions.


Spatial analysis of the influence on "microplastic communities" in the water at a medium scale.
The issue of microplastics in freshwater has been growing in concern. Besides their abundance, the characteristics of microplastics are also important issues. The concept of "microplastic communities" has been utilized to assess differences in microplastic characteristics. In this study, we utilized the "microplastic community" approach to evaluate the impact of land use on microplastic characteristics in water at a provincial scale in China. The abundance of microplastics in water bodies in Hubei Province varied between 0.33 items/L and 5.40 items/L, with an average of 1.74 items/L. Microplastics were significantly more abundant in rivers than in lakes and reservoirs, and their abundance was negatively correlated with the distance from the nearest residential district of sampling sites. Similarities of microplastic communities were significantly different in mountainous and plain areas. Anthropogenic surfaces increased microplastic abundance and tended to decrease the size of microplastics, whereas natural vegetation had the opposite effect. The effect of land use on microplastic community similarity was greater than that of geographic distance. However, spatial scale limits the effect of various factors on microplastic community similarity. This study revealed the comprehensive influence of land use on microplastic characteristics in water and emphasized the importance of spatial scale in the study of microplastic characteristics.


Peritoneal-mediastinal leakage complication of peritoneal dialysis.
The authors report a case of mediastinal fluid collection resulting from peritoneal-mediastinal communication after continuous ambulatory peritoneal dialysis (CAPD). To the best of the authors' knowledge, this is the first reported case in the medical literature. A dry cough developed in the patient who had been receiving CAPD for 4 years. A mediastinal mass owing to peritoneal leakage of dialysate to the mediastinum was confirmed by a computed tomography scan taken 4 hours after the intraperitoneal infusion of contrast-mixed dialysate. The leakage persisted for 12 weeks after the discontinuation of CAPD fluid instillation.


B Vitamin Complex and Chemotherapy-Induced Peripheral Neuropathy.
The purpose of this mini review is to evaluate the literature on B vitamins and chemotherapy-induced peripheral neuropathy. One hundred and five journal articles were evaluated and nine manuscripts were included. There was one in vitro, one was an animal and seven were human studies. The in vitro study was a safety study on vitamin B6 and oxaliplatin which was not directly related to CIPN. The animal study evaluated vitamin B3 on paclitaxel administration with positive results. The human studies varied using a vitamin B complex, vitamin B12 only and vitamin B6. Chemotherapy-induced peripheral neuropathy (CIPN) continues to plague patients and the medical fraternity. Currently, there are still no conclusive protective or treatment options. B vitamins have been found to play a role in CIPN prevention, but further studies are required to ascertain possible protection and treatment options.


Biological characteristics, biosafety prevention and control strategies for the 2022 multi-country outbreak of monkeypox.
Monkeypox is a zoonotic disease caused by the monkeypox virus (MPXV), which is a potential biological warfare agent of bioterrorism and poses the greatest threat to the world's public biosafety and health after variola virus (VARV). While the coronavirus disease 2019 (COVID-19) pandemic has not ended yet, monkeypox is spreading menacingly. The first case of monkeypox in a nonendemic country was confirmed on May 6th, 2022, while the first imported case from Asia was found on June 21st. There were more than 16 thousand reported cases as of July 23rd, the day the World Health Organization (WHO) declared the global monkeypox outbreak a public health emergency of international concern (PHEIC) at the same level as smallpox and COVID-19; while there were more than 53 thousand cases as of September 1st. Therefore, we will propose relevant biosafety prevention and control strategies after analyzing the etiology of the 2022 multi-country monkeypox outbreak from the biological feature, transmissibility, epidemic, and variability of MPXV.