Experimental and Computational Analysis of Newly Identified Pathogenic Mutations in the Creatine Transporter SLC6A8.
Creatine is an essential metabolite for the storage and rapid supply of energy in muscle and nerve cells. In humans, impaired metabolism, transport, and distribution of creatine throughout tissues can cause varying forms of mental disability, also known as creatine deficiency syndrome (CDS). So far, 80 mutations in the creatine transporter (SLC6A8) have been associated to CDS. To better understand the effect of human genetic variants on the physiology of SLC6A8 and their possible impact on CDS, we studied 30 missense variants including 15 variants of unknown significance, two of which are reported here for the first time. We expressed these variants in HEK293 cells and explored their subcellular localization and transport activity. We also applied computational methods to predict variant effect and estimate site-specific changes in thermodynamic stability. To explore variants that might have a differential effect on the transporter's conformers along the transport cycle, we constructed homology models of the inward facing, and outward facing conformations. In addition, we used mass-spectrometry to study proteins that interact with wild type SLC6A8 and five selected variants in HEK293 cells. In silico models of the protein complexes revealed how two variants impact the interaction interface of SLC6A8 with other proteins and how pathogenic variants lead to an enrichment of ER protein partners. Overall, our integrated analysis disambiguates the pathogenicity of 15 variants of unknown significance revealing diverse mechanisms of pathogenicity, including two previously unreported variants obtained from patients suffering from the creatine deficiency syndrome.


Detailed severity assessment of Cincinnati Prehospital Stroke Scale to detect large vessel occlusion in acute ischemic stroke.
Selecting stroke patients with large vessel occlusion (LVO) based on prehospital stroke scales could provide a faster triage and transportation to a comprehensive stroke centre resulting a favourable outcome. We aimed here to explore the detailed severity assessment of Cincinnati Prehospital Stroke Scale (CPSS) to improve its ability to detect LVO in acute ischemic stroke (AIS) patients. A cross-sectional analysis was performed in a prospectively collected registry of consecutive patients with first ever AIS admitted within 6 h after symptom onset. On admission stroke severity was assessed using the National Institutes of Health Stroke Scale (NIHSS) and the presence of LVO was confirmed by computed tomography angiography (CTA) as an endpoint. A detailed version of CPSS (d-CPSS) was designed based on the severity assessment of CPSS items derived from NIHSS. The ability of this scale to confirm an LVO was compared to CPSS and NIHSS respectively. Using a ROC analysis, the AUC value of d-CPSS was significantly higher compared to the AUC value of CPSS itself (0.788 vs. 0.633, p < 0.001) and very similar to the AUC of NIHSS (0.795, p = 0.510). An optimal cut-off score was found as d-CPSS≥5 to discriminate the presence of LVO (sensitivity: 69.9%, specificity: 75.2%). A detailed severity assessment of CPSS items (upper extremity weakness, facial palsy and speech disturbance) could significantly increase the ability of CPSS to discriminate the presence of LVO in AIS patients.


Goodley polyaxial cervical traction. A new approach to a traditional treatment.
A cervical traction system is discussed that offers advantages over traditional methods of applying cervical traction. The major advantage of this system is that it allows precise application of traction to only the restricted spinal segments. Because of this specificity, traction may be applied at reduced force, decreasing the risk of injury. The traction apparatus is adjusted easily to the individual and may be used in the clinic or at home. My experience with the apparatus has shown greater patient compliance, resulting in increased treatment effectiveness, when compared with traditional traction methods. Four representative case studies demonstrate the effectiveness of this traction system. Closely controlled studies are needed to substantiate the effectiveness and safety of this traction system.


Seeking the Roles for Fungal Small-Secreted Proteins in Affecting Saprophytic Lifestyles.
Small secreted proteins (SSPs) comprise 40-60% of the total fungal secretome and are present in fungi of all phylogenetic groups, representing the entire spectrum of lifestyles. They are characteristically shorter than 300 amino acids in length and have a signal peptide. The majority of SSPs are coded by orphan genes, which lack known domains or similarities to known protein sequences. Effectors are a group of SSPs that have been investigated extensively in fungi that interact with living hosts, either pathogens or mutualistic systems. They are involved in suppressing the host defense response and altering its physiology. Here, we aim to delineate some of the potential roles of SSPs in saprotrophic fungi, that have been bioinformatically predicted as effectors, and termed in this mini-review as "effector-like" proteins. The effector-like Ssp1 from the white-rot fungus Pleurotus ostreatus is presented as a case study, and its potential role in regulating the ligninolytic system, secondary metabolism, development, and fruiting body initiation are discussed. We propose that deciphering the nature of effector-like SSPs will contribute to our understanding of development and communication in saprophytic fungi, as well as help, to elucidate the origin, regulation, and mechanisms of fungal-host, fungal-fungal, and fungal-bacterial interactions.


A comparison of end-of-life care quality for Veterans receiving hospice in VA nursing homes and community nursing homes.
While the Veterans Health Administration (VA) has long provided hospice care within VA community living centers (CLCs, i.e., VA nursing homes), an increasing number of Veterans are receiving hospice in VA-contracted community nursing homes (CNHs). However, little data exist about the quality of end-of-life (EOL) care provided in CNHs. The aim of this study was to compare family ratings of the quality of EOL care provided to Veterans receiving hospice in VA CLCs and VA-contracted CNHs. We conducted a retrospective analysis of national data from VA's electronic medical record and Bereaved Family Survey (BFS) for Veterans who received hospice in VA CLCs or VA-contracted CNHs between October 2021 and March 2022. The final sample included 1238 Veterans who died in either a CLC (n = 1012) or a CNH (n = 226) and whose next-of-kin completed the BFS. Our primary outcome was the BFS global rating of care received in the last 30 days of life. Secondary outcomes included BFS items related to symptom management, communication, emotional and spiritual support, and information about burial and survivor benefits. We compared unadjusted and adjusted proportions for all BFS outcomes between those who received hospice in CLCs and CNHs. The adjusted proportion of family members who gave the best possible rating (a score of 9 or 10 out of a possible 10) for the overall care received near EOL was more than 13 percentage points higher for Veterans who received hospice in VA CLCs compared to VA-contracted CNHs. Our findings also revealed quality gaps of even greater magnitude in specific EOL care-focused domains. Our findings document inadequacies in the quality of multiple aspects of EOL care provided to Veterans in CNH-based hospice and illuminate the urgent need for policy and practice interventions to improve this care.


Assessment of sexual dysfunction in patients with multiple sclerosis: a perspective from neurologist.
Sexual dysfunction (SD) is a common comorbidity in people with multiple sclerosis (pwMS). It affects the quality of life and remains an overlooked condition. The objective of this study was to describe how Colombian neurologists assess and treat SD and explore the barriers during sexual function evaluation. In this observational cross-sectional study we developed a questionnaire for neurologists with 4 sections (demographic data, evaluation and treatment of SD, and possible reasons for not discussing sexual dysfunction.) It was sent via email to 326 Colombian neurologists. We grouped the answers according to the type of consultation (neurologists from a MS program or no MS program). We described through absolute frequencies and proportions. Fifty neurologists answered the survey. 64% stated that they usually study sexual dysfunction in neurological disorders. The main methods employed were private reading (86%) and attending conferences (14%). 5/50 participants have never attend pwMS; the Sect. 2-4 was not answered by them. 29% work in a MS program, all of them asked their patients about sexual function, but 18.75% of physicians working outside an MS program have never asked about it. Main reasons for not talking about sexual dysfunction were lack of knowledge (65.1%), presence of a companion (65.1%) and lack of time (55.8%). 91% of the neurologists reported that their patients usually and frequently ask about sexual function. Neurologists use informal questions to assess sexual function (80%), although 64.4% said that they are aware of SD questionnaires. When sexual dysfunction is detected, 91% of neurologists refer patients to another specialist and 87% do not start any treatment. Colombian neurologists are concerned with sexual function in pwMS, however it remains an underdiagnosed an undertreated condition. It is necessary to strengthen knowledge about the diagnosis and treatment of sexual dysfunction in pwMS, for neurologists and patients. It is also imperative to eliminate barriers around the topic and include sexual function evaluation and treatment as the routine care of pwMS.


[Phase I study of alizapride in cancer patients treated with cisplatin].
Alizapride in a new methoxy-2 benzamide derivative with antiemetic properties. The dose-effect relationship of this compound was studied in 24 cancer patients treated with cisplatin regimens. Alizapride was administered as 15 min intravenous infusions at doses of 1 to 5 mg/kg repeated every 2 hours for 5 administrations. No adverse reactions were noted at doses of 1 to 3 mg/kg. Serious orthostatic hypotension was observed at the highest dose level. Other side effects were essentially mild to moderate and consisted of somnolence and diarrhea. The study indicated that the dosage level of 4 mg/kg is suitable for clinical use. The antiemetic activity of alizapride in patients receiving cisplatin therapy remains to be firmly established in a randomized study. This trial suggested a better antiemetic effect of alizapride in naive patients as compared to those previously treated with anticancer chemotherapy.


Action plans used in action observation.
How do we understand the actions of others? According to the direct matching hypothesis, action understanding results from a mechanism that maps an observed action onto motor representations of that action. Although supported by neurophysiological and brain-imaging studies, direct evidence for this hypothesis is sparse. In visually guided actions, task-specific proactive eye movements are crucial for planning and control. Because the eyes are free to move when observing such actions, the direct matching hypothesis predicts that subjects should produce eye movements similar to those produced when they perform the tasks. If an observer analyses action through purely visual means, however, eye movements will be linked reactively to the observed action. Here we show that when subjects observe a block stacking task, the coordination between their gaze and the actor's hand is predictive, rather than reactive, and is highly similar to the gaze-hand coordination when they perform the task themselves. These results indicate that during action observation subjects implement eye motor programs directed by motor representations of manual actions and thus provide strong evidence for the direct matching hypothesis.


Alkylaminophenol and GPR17 Agonist for Glioblastoma Therapy: A Combinational Approach for Enhanced Cell Death Activity.
Drug resistance and tumor heterogeneity limits the therapeutic efficacy in treating glioblastoma, an aggressive infiltrative type of brain tumor. GBM cells develops resistance against chemotherapeutic agent, temozolomide (TMZ), which leads to the failure in treatment strategies. This enduring challenge of GBM drug resistance could be rational by combinatorial targeted therapy. Here, we evaluated the combinatorial effect of phenolic compound (2-(3,4-dihydroquinolin-1(2H)-yl)(p-tolyl)methyl)phenol (THTMP), GPR17 agonist 2-({5-[3-(Morpholine-4-sulfonyl)phenyl]-4-[4-(trifluoromethoxy)phenyl]-4H-1,2,4-triazol-3-yl}sulfanyl)-N-[4-(propan-2-yl)phenyl]acetamide (T0510.3657 or T0) with the frontline drug, TMZ, on the inhibition of GBM cells. Mesenchymal cell lines derived from patients' tumors, MMK1 and JK2 were treated with the combination of THTMP + T0, THTMP + TMZ and T0 + TMZ. Cellular migration, invasion and clonogenicity assays were performed to check the migratory behavior and the ability to form colony of GBM cells. Mitochondrial membrane permeability (MMP) assay and intracellular calcium, [Ca2+]i, assay was done to comprehend the mechanism of apoptosis. Role of apoptosis-related signaling molecules was analyzed in the induction of programmed cell death. In vivo validation in the xenograft models further validates the preclinical efficacy of the combinatorial drug. GBM cells exert better synergistic effect when exposed to the cytotoxic concentration of THTMP + T0, than other combinations. It also inhibited tumor cell proliferation, migration, invasion, colony-forming ability and cell cycle progression in S phase, better than the other combinations. Moreover, the combination of THTMP + T0 profoundly increased the [Ca2+]i, reactive oxygen species in a time-dependent manner, thus affecting MMP and leading to apoptosis. The activation of intrinsic apoptotic pathway was regulated by the expression of Bcl-2, cleaved caspases-3, cytochrome c, HSP27, cIAP-1, cIAP-2, p53, and XIAP. The combinatorial drug showed promising anti-tumor efficacy in GBM xenograft model by reducing the tumor volume, suggesting it as an alternative drug to TMZ. Our findings indicate the coordinated administration of THTMP + T0 as an efficient therapy for inhibiting GBM cell proliferation.


Calcofluor white combination antifungal treatments for Trichophyton rubrum and Candida albicans.
Superficial mycoses caused by dermatophyte fungi are among the most common infections worldwide, yet treatment is restricted by limited effective drugs available, drug toxicity, and emergence of drug resistance. The stilbene fluorescent brightener calcofluor white (CFW) inhibits fungi by binding chitin in the cell wall, disrupting cell wall integrity, and thus entails a different mechanism of inhibition than currently available antifungal drugs. To identify novel therapeutic options for the treatment of skin infections, we compared the sensitivity of representative strains of the dermatophyte Trichophyton rubrum and Candida albicans to CFW and a panel of fluorescent brighteners and phytoalexin compounds. We identified the structurally related stilbene fluorescent brighteners 71, 85, 113 and 134 as fungicidal to both T. rubrum and C. albicans to a similar degree as CFW, and the stilbene phytoalexins pinosylvan monomethyl ether and pterostilbene inhibited to a lesser degree, allowing us to develop a structure-activity relationship for fungal inhibition. Given the abilities of CFW to absorb UV(365 nm) and bind specifically to fungal cell walls, we tested whether CFW combined with UV(365 nm) irradiation would be synergistic to fungi and provide a novel photodynamic treatment option. However, while both treatments individually were cytocidal, UV(365 nm) irradiation reduced sensitivity to CFW, which we attribute to CFW photoinactivation. We also tested combination treatments of CFW with other fungal inhibitors and identified synergistic interactions between CFW and some ergosterol biosynthesis inhibitors in C. albicans. Therefore, our studies identify novel fungal inhibitors and drug interactions, offering promise for combination topical treatment regimes for superficial mycoses.


Focused Ultrasound Stimulates the Prefrontal Cortex and Prevents MK-801-Induced Psychiatric Symptoms of Schizophrenia in Rats.
Treatment of schizophrenia remains a major challenge. Recent studies have focused on glutamatergic signaling hypoactivity through N-methyl-D-aspartate (NMDA) receptors. Low-intensity pulsed ultrasound (LIPUS) improves behavioral deficits and ameliorates neuropathology in dizocilpine (MK-801)-treated rats. The aim of this study was to investigate the efficacy of LIPUS against psychiatric symptoms and anxiety-like behaviors. Rats assigned to 4 groups were pretreated with or without LIPUS for 5 days. The open field and prepulse inhibition tests were performed after saline or MK-801 (0.3 mg/kg) administration. Then, the neuroprotective effects of LIPUS on the MK-801-treated rats were evaluated using western blotting and immunohistochemical staining. LIPUS stimulation of the prefrontal cortex (PFC) prevented deficits in locomotor activity and sensorimotor gating and improved anxiety-like behavior. MK-801 downregulated the expression of NR1, the NMDA receptor, in rat medial PFC (mPFC). NR1 expression was significantly higher in animals receiving LIPUS pretreatment compared to those receiving only MK-801. In contrast, a significant increase in c-Fos-positive cells in the mPFC and ventral tegmental area was observed in the MK-801-treated rats compared to those receiving only saline; this change was suppressed by pretreatment with LIPUS. This study provides new evidence for the role of LIPUS stimulation in regulating the NMDA receptor and modulating c-Fos activity, which makes it a potentially valuable antipsychotic treatment for schizophrenia.


Computing the joint distribution of the total tree length across loci in populations with variable size.
In recent years, a number of methods have been developed to infer complex demographic histories, especially historical population size changes, from genomic sequence data. Coalescent Hidden Markov Models have proven to be particularly useful for this type of inference. Due to the Markovian structure of these models, an essential building block is the joint distribution of local genealogical trees, or statistics of these genealogies, at two neighboring loci in populations of variable size. Here, we present a novel method to compute the marginal and the joint distribution of the total length of the genealogical trees at two loci separated by at most one recombination event for samples of arbitrary size. To our knowledge, no method to compute these distributions has been presented in the literature to date. We show that they can be obtained from the solution of certain hyperbolic systems of partial differential equations. We present a numerical algorithm, based on the method of characteristics, that can be used to efficiently and accurately solve these systems and compute the marginal and the joint distributions. We demonstrate its utility to study the properties of the joint distribution. Our flexible method can be straightforwardly extended to handle an arbitrary fixed number of recombination events, to include the distributions of other statistics of the genealogies as well, and can also be applied in structured populations.


Probing the electronic structure and chemical bonding of gold oxides and sulfides in AuOn(-) and AuSn(-) (n = 1, 2).
The Au-O and Au-S interactions are essential in nanogold catalysis and nanotechnology, for which monogold oxide and sulfide clusters serve as the simplest molecular models. We report a combined photoelectron spectroscopy and ab initio study on AuO (-) and AuO 2 (-) and their valent isoelectronic AuS (-) and AuS 2 (-) species to probe their electronic structure and to elucidate the Au-O and Au-S chemical bonding. Vibrationally resolved spectra were obtained at different photon energies, providing a wealth of electronic structure information for each species. Similar spectra were observed for AuO (-) and AuS (-) and for the linear OAuO (-) and SAuS (-) species. A bent isomer was also observed as Au(S 2) (-) in the AuS 2 (-) spectra, whereas a similar Au(O 2) (-) complex was not observed in the case of AuO 2 (-). High-level ab initio calculations were conducted to aid spectral assignments and provide insight into the chemical bonding in the AuX (-) and AuX 2 (-) molecules. Excellent agreement is achieved between the calculated electronic excitations and the observed spectra. Configuration interactions and spin-orbit couplings were shown to be important and were necessary to achieve good agreement between theory and experiment. Strong covalent bonding was found in both the AuX (-) and the XAuX (-) species with multiple bonding characters. While Au(S 2) (-) was found to be a low-lying isomer with a significant binding energy, Au(O 2) (-) was shown to be unbound consistent with the experimental observation. The latter is understood in the context of the size-dependent reactivity of Au n (-) clusters with O 2.


Predicting lung cancer prior to surgical resection in patients with lung nodules.
Existing predictive models for lung cancer focus on improving screening or referral for biopsy in general medical populations. A predictive model calibrated for use during preoperative evaluation of suspicious lung lesions is needed to reduce unnecessary operations for a benign disease. A clinical prediction model (Thoracic Research Evaluation And Treatment [TREAT]) is proposed for this purpose. We developed and internally validated a clinical prediction model for lung cancer in a prospective cohort evaluated at our institution. Best statistical practices were used to construct, evaluate, and validate the logistic regression model in the presence of missing covariate data using bootstrap and optimism corrected techniques. The TREAT model was externally validated in a retrospectively collected Veteran Affairs population. The discrimination and calibration of the model was estimated and compared with the Mayo Clinic model in both the populations. The TREAT model was developed in 492 patients from Vanderbilt whose lung cancer prevalence was 72% and validated among 226 Veteran Affairs patients with a lung cancer prevalence of 93%. In the development cohort, the area under the receiver operating curve (AUC) and Brier score were 0.87 (95% confidence interval [CI], 0.83-0.92) and 0.12, respectively compared with the AUC 0.89 (95% CI, 0.79-0.98) and Brier score 0.13 in the validation dataset. The TREAT model had significantly higher accuracy (p < 0.001) and better calibration than the Mayo Clinic model (AUC = 0.80; 95% CI, 75-85; Brier score = 0.17). The validated TREAT model had better diagnostic accuracy than the Mayo Clinic model in preoperative assessment of suspicious lung lesions in a population being evaluated for lung resection.


Sugammadex: another milestone in clinical neuromuscular pharmacology.
Sugammadex is a revolutionary investigational reversal drug currently undergoing Phase III testing whose introduction into clinical practice may change the face of clinical neuromuscular pharmacology. A modified gamma-cyclodextrin, sugammadex exerts its effect by forming very tight water-soluble complexes at a 1:1 ratio with steroidal neuromuscular blocking drugs (rocuronium > vecuronium >> pancuronium). During rocuronium-induced neuromuscular blockade, the IV administration of sugammadex creates a concentration gradient favoring the movement of rocuronium molecules from the neuromuscular junction back into the plasma, which results in a fast recovery of neuromuscular function. Sugammadex is biologically inactive, does not bind to plasma proteins, and appears to be safe and well tolerated. Additionally, it has no effect on acetylcholinesterase or any receptor system in the body. The compound's efficacy as an antagonist does not appear to rely on renal excretion of the cyclodextrin-relaxant complex. Human and animal studies have demonstrated that sugammadex can reverse very deep neuromuscular blockade induced by rocuronium without muscle weakness. Its future clinical use should decrease the incidence of postoperative muscle weakness, and thus contribute to increased patient safety. Sugammadex will also facilitate the use of rocuronium for rapid sequence induction of anesthesia by providing a faster onset-offset profile than that seen with 1.0 mg/kg succinylcholine. Furthermore, no additional anticholinesterase or anticholinergic drugs would be needed for antagonism of residual neuromuscular blockade, which would mean the end of the cardiovascular and other side effects of these compounds. The clinical use of sugammadex promises to eliminate many of the shortcomings in our current practice with regard to the antagonism of rocuronium and possibly other steroidal neuromuscular blockers.


Continuous vs. interval training: a review for the athlete and the coach.
Both interval and continuous training are necessary in order to maximize the endurance athlete's potential for competition. Continuous submaximal training exerts its greatest effect upon the oxygen transport system of the body, while high intensity endurance-interval training exerts its greatest effect on the structural and biochemical properties of the muscle. The physiological basis for both forms of training is discussed and recommendations are made for year-round training and training to elevate the anaerobic threshold.


Effects of scatter and attenuation correction on quantitative analysis of beta-CIT brain SPET.
The aim of this study was to evaluate the effects of scatter correction (SC) and attenuation correction (AC) on the quantification of dopamine transporters using 123I-beta-CIT brain SPET images. Quantitative analysis was carried out using static SPET images obtained 23 h after injection. We calculated V3" [(striatal-occipital)/occipital ratio] values from images without correction, with AC, and with SC and AC. Two types of regions of interest (ROI) were placed on the striatum: a small square ROI and a larger ROI containing most of the striatum. After validating the correction method in a phantom experiment, a human study was carried out involving eight normal volunteers and 15 patients. The larger ROI yielded smaller V3" values. The effect of attenuation correction was modest, whereas that of scatter correction was marked. It was shown that beta-CIT SPET quantification was affected by the size of the ROI, photon scattering and attenuation, and that scatter and attenuation correction improved the accuracy of the quantification. Methodological standardization in image processing and the type of ROI should be considered when a multi-centre trial is planned.


Rat mammary-gland acetyl-coenzyme A carboxylase interaction with milk fatty acids.
1. Highly purified rat mammary-gland acetyl-CoA carboxylase was inhibited by milk obtained from rats 12h after their young were weaned. 2. All the inhibitory activity was found in the particulate fraction (R(105)) obtained on centrifuging the milk. It could be extracted from milk fraction R(105) with acetone and identified as a complex mixture of non-esterified fatty acids, present in high concentration (nearly 10mm) in the milk. 3. Inhibition of acetyl-CoA carboxylase was observed at low concentrations (0.2-20mum) of several of these fatty acids when fresh fully active enzyme was used. Enzyme that had been partly inactivated by aging, or by storing in the absence of citrate, was stimulated by low concentrations but inhibited by high concentrations of fatty acids. 4. Various experiments suggested that fatty acids produce irreversible inactivation of acetyl-CoA carboxylase. 5. The effects of palmitoyl-CoA on mammary-gland acetyl-CoA carboxylase were found to resemble those of fatty acids, except that palmitoyl-CoA was effective at lower concentration. 6. The effect of milk fraction R(105) was tested on six other enzymes previously shown to decline to various extents after weaning. Although several of these enzymes were affected by unfractionated milk fraction R(105), none was significantly inhibited by the acetone extract or by low concentrations of lauric acid. 7. The findings are consistent, both qualitatively and quantitatively, with a regulatory mechanism whereby milk fatty acids shut off fatty acid synthesis in the mammary gland after weaning by inhibiting acetyl-CoA carboxylase.


Development of a bead-based multiplex immunoassay for simultaneous quantitative detection of IgG serum antibodies against seven vaccine-preventable diseases.
Serosurveillance and seroprevalence studies should be carried out to monitor vaccine-preventable diseases. Multiplex immunoassay (MIA) systems are useful tools for this purpose, allowing the simultaneous quantitative detection of antibodies in one small serum sample, which presents an advantage over conventional methods, such as enzyme-linked immunosorbent assays (ELISAs). Therefore, we developed a multiplex immunoassay for the measurement of antibodies against seven vaccine-preventable infections (measles, rubella, mumps, tetanus, diphtheria, pertussis and Haemophilus influenza type b (Hib) infection). In our multiplex system, heterologous inhibition generally did not exceed 10%, while homologous inhibition varied between 90 and 98%. The intra- and inter-assay variability was ≤11%. The results of in-house MIA showed satisfactory correlation with commercial ELISAs, with Spearman correlation coefficients from 0.90 to 0.98. At the cut-off values defined for our MIA the serostatus can be determined with high sensitivity (89-100%) and specificity (92-98%). Thus, the developed in-house MIA represents a feasible alternative to conventional ELISAs and could be used for large-scale serosurveillance/seroprevalence studies of vaccine-preventable diseases.


Proopiocortin-related peptide plasma levels throughout prepuberty and puberty.
The plasma patterns of ACTH, beta-lipotropin (beta LPH) and beta-endorphin (beta EP), in addition to those of cortisol and dehydroepiandrosterone sulfate (DHAS), were studied in 139 prepubertal children (subdivided into different age groups) and 38 adolescents (subdivided according to Tanner's pubertal stages) aged 10-16 yr. The adult control group was composed of 23 females and 12 males aged 17-40 yr. No sex differences were found in ACTH, beta LPH, beta EP, and cortisol plasma levels. ACTH plasma levels were slightly lower in the 1- to 3-yr-old groups than in males at 4-5 yr and females at 8-9 yr. No further significant differences were observed in any of the age or pubertal groups, the concentrations being constantly in the adult range. beta LPH and beta EP plasma levels were lowest at 1-3 yr in both males (beta LPH: 2.1 +/- 0.25, beta EP: 1.85 +/- 0.59 fmol/ml, mean +/- SE) and females (beta LPH: 2.8 +/- 0.31; beta EP: 2.41 +/- 0.41 fmol/ml); plasma levels of both hormones increased progressively in both sexes until Puberty 1 stage of sexual maturation, at which time levels were 7.3 +/- 0.78 and 8.69 +/- 1.0 fmol/ml in males and 7.1 +/- 0.34 and 6.76 +/- 0.13 fmol/ml in females; these levels are similar to adult values. A highly significant linear correlation was found between both beta LPH and beta EP concentrations and the age of the subjects; this was not true for ACTH plasma levels. Cortisol plasma levels were similar in all groups. DHAS plasma levels increased progressively from 1-3 yr to the end of sexual maturation when adult values were reached. During prepuberty, DHAS levels were significantly correlated with both beta LPH and beta EP, but not ACTH. These data indicate that plasma beta LPH and beta EP concentrations, in contrast to ACTH levels, increase progressively throughout prepuberty and suggest that the processing of the parent proopiocortin molecule or secretion of the processed peptides from the anterior pituitary (or other sources) may change from early infancy to adulthood. Furthermore, the correlation between both beta LPH and beta EP with DHAS plasma levels in prepuberty suggests a role of proopiocortin-related peptides in adrenarche.


ContinuousFlex: Software package for analyzing continuous conformational variability of macromolecules in cryo electron microscopy and tomography data.
ContinuousFlex is a user-friendly open-source software package for analyzing continuous conformational variability of macromolecules in cryo electron microscopy (cryo-EM) and cryo electron tomography (cryo-ET) data. In 2019, ContinuousFlex became available as a plugin for Scipion, an image processing software package extensively used in the cryo-EM field. Currently, ContinuousFlex contains software for running (1) recently published methods HEMNMA-3D, TomoFlow, and NMMD; (2) earlier published methods HEMNMA and StructMap; and (3) methods for simulating cryo-EM and cryo-ET data with conformational variability and methods for data preprocessing. It also includes external software for molecular dynamics simulation (GENESIS) and normal mode analysis (ElNemo), used in some of the mentioned methods. The HEMNMA software has been presented in the past, but not the software of other methods. Besides, ContinuousFlex currently also offers a deep learning extension of HEMNMA, named DeepHEMNMA. In this article, we review these methods in the context of the ContinuousFlex package, developed to facilitate their use by the community.


A resilience-based robustness evaluation framework for sustainable urban flood management under uncertainty.
Urban drainage systems (UDSs) may experience failure encountering uncertain future conditions. These uncertainties arise from internal and external threats such as sedimentation, blockage, and climate change. In this paper, a new resilience-based framework is proposed to assess the robustness of urban flood management strategies under some distinct future scenarios. The robustness values of flood management strategies are evaluated by considering reliability, resiliency, and socio-ecological resilience criteria. The socio-ecologic resilience criteria are proposed considering the seven principles of building resilience proposed by Biggs et al. (2012). The evidential reasoning (ER) approach and the regret theory are utilized to calculate the total robustness of the flood management strategies. In this framework, the non-dominated sorting genetic algorithms III (NSGA-III) optimization model and the storm water management model (SWMM) simulation model are linked and run to quantify the criteria. The novelty of this paper lies in presenting a new framework to increase the sustainability and resilience of cities against floods considering the deep uncertainties in the main economic, social, and hydrological factors. This methodology provides policies for redesigning and sustainable operation of urban infrastructures to deal with floods. To evaluate the applicability and efficiency of the framework, it is applied to the East drainage catchment of the Tehran metropolitan area in Iran. The results show that real-time operation of existing flood detention reservoirs, along with implementing five new relief tunnels with a construction cost of 37.1 million dollars, is the most robust non-dominated strategy for flood management in the study area. Comparing the results of the proposed framework with those of a traditional framework shows that it can increase the robustness value by about 40% with the same implementation cost.


Pesticide occurrence in a subtropical estuary, Australia: Complementary sampling methods.
Monitoring pesticide run-off in the aquatic environment is ecologically important. Effective methods are required to detect the wide range of possible pesticides that enter estuaries from the surrounding catchment. Here, we investigate the occurrence of pesticides in the Richmond River estuary, Australia, and compare the effectiveness of using oysters and Chemcatcher® passive sampling devices against composite water samples. Samples were collected from six sites during two sampling periods: from January to March 2020 (4 weekly composite water samples and oyster collections) and from February to March 2021 (8 twice weekly composite water samples and Chemcatcher® deployment). Samples were analysed for up to 174 pesticides. A total of 21 pesticides were detected across all sites using all methods. The number of pesticides and mixture of pesticides detected in the 2020 sampling was higher in oyster samples than in water samples. In 2021, Chemcatcher® samplers detected more pesticides than in water samples. Herbicides were the most common in all samples. Insecticides and most fungicides were detected only in oysters and Chemcatcher®. Overall, the use of three complementary sampling approaches demonstrated a high level of pesticide input into the Richmond River estuary, highlighting the usefulness of oysters as biomonitors for some pesticides.


[The clinical value of air sonography in the diagnosis of meniscus lesions. A prospective study of 50 knee joints].
This paper shows the results of using air as a contrast medium in ultrasonography of the menisci (aerosonography). 100 menisci (50 knees) were examined with aerosonography and in 93 = 93% the findings were proven to be true by subsequent arthroscopy. 6% of the findings were false positive (2 medial menisci and 4 lateral menisci). One false negative finding concerned a lesion at the free inner edge of the lateral meniscus. Not one single lesion of the medial meniscus was totally missed. The sensitivity of aerosonography is 97.8%, the specificity 89.1%. The predictive value is 88%, the accuracy is 93%. Considering the medial meniscus alone the aerosonographic diagnosis proved to be true in 96% (sensitivity for the medial meniscus 100%, specificity 87.5%, predictive value 94.4%, accuracy 96%). 90% of the aerosonographic diagnosis concerning the lateral meniscus were confirmed by arthroscopy (sensitivity for the lateral meniscus 90.9%, specificity 89.1%, predictive value 71.4%, accuracy 90%).


Assessing the Clinical, Economic, and Health Resource Utilization Impacts of Prefilled Syringes Versus Conventional Medication Administration Methods: Results From a Systematic Literature Review.
The objective of this systematic review was to assess the clinical, economic, and health resource utilization outcomes associated with the use of prefilled syringes in medication administration compared with traditional preparation methods. We conducted a systematic literature review to evaluate outcomes such as medication errors, wastage, time savings, and contamination in prefilled syringes. Our search encompassed multiple databases, including PubMed and Embase, for studies published between January 1, 2017, and November 1, 2022. Peer-reviewed publications meeting our inclusion criteria underwent rigorous screening, including title, abstract, and full-text article assessments, performed by two reviewers. Among reviewed articles, 24 met our eligibility criteria. Selected studies were primarily observational (46%) and conducted in Europe (46%). Our findings indicated that prefilled syringes consistently reduced medication errors (by 10%-73%), adverse events (from 1.1 to 0.275 per 100 administrations), wastage (by up to 80% of drug), and preparation time (from 4.0 to 338.0 seconds) (ranges varied by drug type, setting, and dosage). However, there was limited data on contamination. Economically, prefilled syringes reduced waste and error rates, which may translate into overall savings. This review highlights the value of prefilled syringes, which can streamline medication delivery, save nursing time, and reduce preventable medication errors. Moreover, prefilled syringes have the potential to minimize medication wastage, optimizing resource utilization and efficiency in health care settings. Our findings provide new insights into clinical and economic benefits of prefilled syringe adoption. These benefits include improved medication delivery and safety, which can lead to time and cost reductions for health care departments, hospitals, and health systems. However, further real-world research on clinical and economic outcomes, especially in contamination, is needed to better understand the benefits of prefilled syringes.


Function approximation based on fuzzy rules extracted from partitioned numerical data.
We present an efficient method for extracting fuzzy rules directly from numerical input-output data for function approximation problems. First, we convert a given function approximation problem into a pattern classification problem. This is done by dividing the universe of discourse of the output variable into multiple intervals, each regarded as a class, and then by assigning a class to each of the training data according to the desired value of the output variable. Next, we partition the data of each class in the input space to achieve a higher accuracy in approximation of class regions. Partition terminates according to a given criterion to prevent excessive partition. For class region approximation, we discuss two different types of representations using hyperboxes and ellipsoidal regions, respectively. Based on a selected representation, we then extract fuzzy rules from the approximated class regions. For a given input datum, we convert, or in other words, defuzzify, the resulting vector of the class membership degrees into a single real value. This value represents the final result approximated by the method. We test the presented method on a synthetic nonlinear function approximation problem and a real-world problem in an application to a water purification plant. We also compare the presented method with a method based on neural networks.


Real-time observation of bimodal proton transfer in acid-base pairs in water.
The neutralization reaction between an acid and a base in water, triggered after optical excitation, was studied by femtosecond vibrational spectroscopy. Bimodal dynamics were observed. In hydrogen-bonded acid-base complexes, the proton transfer proceeds extremely fast (within 150 femtoseconds). In encounter pairs formed by diffusion of uncomplexed photoacid and base molecules, the reaction upon contact was an order of magnitude slower, in agreement with earlier reported values. These results call for a refinement of the traditional Eigen-Weller picture of acid-base reactions: A three-stage model is introduced to account for all observed dynamics.


[11C]raclopride-PET studies of the Huntington's disease rate of progression: relevance of the trinucleotide repeat length.
We used [11C]raclopride and positron emission tomography (PET) to assess the relationship between striatal dopamine D2 receptor binding, trinucleotide repeat number (CAG), and subject age in 10 asymptomatic and 8 symptomatic carriers of the Huntington's disease (HD) mutation. In both preclinical and symptomatic gene carriers, we found significant correlations between CAG repeat length and the ratio of percent loss in striatal D2 receptor binding divided by age. In accord with neuropathological studies, we obtained an intercept at 35.5 CAG repeats in the symptomatic HD patients. Nonetheless, we noted that the slopes of the correlation lines differed significantly for the presymptomatic and symptomatic cohorts. These PET results support the notion that the HD disease process is a function of trinucleotide length and age, and that the development of clinical signs and symptoms is associated with CAG repeat lengths greater than 35.5. However, our analysis also suggests that striatal degeneration may proceed in a nonlinear fashion. These findings have implications for the design of neuroprotective strategies for the treatment of HD.


Comparing the loss of functional independence of older adults in the U.S. and China.
Functional loss among older adults is known to follow a hierarchical sequence, but little is known about whether such sequences differ across socio-cultural contexts. The aim of this study is to construct activities of daily livings (ADL) scales for oldest-old adults in the United States and China so as to compare their functional loss sequences. We use data from the Asset and Health Dynamics of the Oldest Old (n=1607) and Chinese Longitudinal Healthy Longevity Survey (n=5570) for years 1998-2008. ADL items are calibrated within a scale using the Rasch measurement model. Rasch scores are averaged across survey waves to identify the ADL loss sequence for each study population. We also assess scale stability over measurement periods. Factor analyses confirm that the ADL items in each study population can be combined meaningfully to form a hierarchical sequence. Internal consistency assessed by Cronbach's alpha is high (0.81 to 0.95). We find that bathing is the first activity that both older Americans and Chinese have difficulty with, while eating is the last activity. There are, however, differences in the rank order for toileting (ranked more challenging in the Chinese sample) and dressing (ranked more challenging in the U.S. sample). Item orderings are stable over time. The results highlight the relative importance of bathing in the functional loss sequence for older adults, regardless of socio-cultural context. Health interventions are needed to address deficits in the bathroom environment, especially in developing countries like China.


Trimming and Washing of Beef Carcasses as a Method of Improving the Microbiological Quality of Meat.
A study to compare procedures and interventions for removing physical and bacterial contamination from beef carcasses was conducted in six carcass conversion operations that were representative of modern, high-volume plants and located in five different states. Treatment procedures included trimming, washing, and the current industry practice of trimming followed by washing. In addition, hot (74 to 87.8°C at the pipe) water washing and rinsing with ozone (0.3 to 2.3 ppm) or hydrogen peroxide (5%) were applied as intervention treatments. Beef carcasses were deliberately contaminated with bovine fecal material at >4.0 log colony-forming units (CFU)/cm2 in order to be better able to observe the decontaminating effects of the treatments. Carcasses were visually scored by 2 to 3 trained personnel for the level of gross contamination before and after treatment. Samples (10 by 15 cm, 0.3 to 0.5 cm thick) for microbiological testing were excised as controls or after application of each procedure or intervention and analyzed for aerobic mesophilic plate counts, Escherichia coli Biotype I counts, and presence or absence of Listeria spp., Salmonella spp., and Escherichia coli O157:H7. Average reductions in aerobic plate counts were 1.85 and 2.00 log CFU/cm2 for the treatments of trimming-washing and hot-water washing, respectively. Hydrogen peroxide and ozone reduced aerobic plate counts by 1.14 and 1.30 log CFU/cm2, respectively. In general, trimming and washing of beef carcasses consistently resulted in low bacterial populations and scores for visible contamination. However, the data also indicated that hot- (74 to 87.8°C at the pipe) water washing was an effective intervention that reduced bacterial and fecal contamination in a consistent manner.


Continuous drug delivery in Parkinson's disease.
Development of motor and non-motor complications during the course of Parkinson's disease (PD) is a major challenge for therapeutic management. At advanced disease stages, patients frequently fluctuate between PD symptoms-such as bradykinesia-and dyskinesias, in response to fluctuations in drug concentrations. Continuous subcutaneous infusion of the dopamine agonist apomorphine or intestinal infusion of levodopa reduce such fluctuations in both pharmacokinetics and motor function. This is the basis for the concept of continuous drug delivery in PD, and the more theoretical concept of continuous dopaminergic stimulation. These expressions are sometimes used to describe a treatment that is more continuous in its pharmacokinetic profile or that produces more sustained effects, compared with immediate-release levodopa, i.e. not only pump treatments. For example, sustained-release formulations of levodopa or dopamine agonists, transdermal delivery of rotigotine, and addition of catechol-O-methyltransferase inhibitors or monoamine oxidase-B inhibitors have been developed with the aim to provide more continuous drug concentrations, sustained benefits and minimized side effects. Progress has been made, but there are still knowledge gaps regarding how these treatment alternatives can be optimally used. New treatments are currently being developed to provide the continuous drug delivery that is known to successfully alleviate motor and non-motor complications. Hopefully, although not yet proven, these new methods may also prevent or postpone some of the late-stage complications.


Total radical yields from tropospheric ethene ozonolysis.
The gas-phase reactions of ozone with alkenes can be significant sources of free radicals (OH, HO(2) and RO(2)) in the Earth's atmosphere. In this study the total radical production and degradation products from ethene ozonolysis have been measured, under conditions relevant to the troposphere, during a series of detailed simulation chamber experiments. Experiments were carried out in the European photoreactor EUPHORE (Valencia, Spain), utilising various instrumentation including a chemical-ionisation-reaction time-of-flight mass-spectrometer (CIR-TOF-MS) measuring volatile organic compounds/oxygenated volatile organic compounds (VOCs/OVOCs), a laser induced fluorescence (LIF) system for measuring HO(2) radical products and a peroxy radical chemical amplification (PERCA) instrument measuring HO(2) + ΣRO(2). The ethene + ozone reaction system was investigated with and without an OH radical scavenger, in order to suppress side reactions. Radical concentrations were measured under dry and humid conditions and interpreted through detailed chemical chamber box modelling, incorporating the Master Chemical Mechanism (MCMv3.1) degradation scheme for ethene, which was updated to include a more explicit representation of the ethene-ozone reaction mechanism.The rate coefficient for the ethene + ozone reaction was measured to be (1.45 ± 0.25) × 10(-18) cm(3) molecules(-1) s(-1) at 298 K, and a stabilised Criegee intermediate yield of 0.54 ± 0.12 was determined from excess CO scavenger experiments. An OH radical yield of 0.17 ± 0.09 was determined using a cyclohexane scavenger approach, by monitoring the formation of the OH-initiated cyclohexane oxidation products and HO(2). The results highlight the importance of knowing the [HO(2)] (particularly under alkene limited conditions and high [O(3)]) and scavenger chemistry when deriving radical yields. An averaged HO(2) yield of 0.27 ± 0.07 was determined by LIF/model fitting. The observed yields are interpreted in terms of branching ratios for each channel within the postulated ethene ozonolysis mechanism.


Autologous micro-fragmented adipose tissue for the treatment of diffuse degenerative knee osteoarthritis: an update at 3 year follow-up.
Conservative therapies for the treatment of knee degenerative processes are used before resorting to surgery; nonetheless, they may offer only short-term benefits. Encouraging preliminary results have been reported using mesenchymal stem cells (MSCs), either alone or in association with surgery. Among the many sources, adipose tissue has created a huge interest, because of its anti-inflammatory and regenerative properties ascribed to the cells of its stromal vascular fraction. We previously reported the safety and feasibility of autologous micro-fragmented adipose tissue as adjuvant for the surgical treatment of diffuse degenerative chondral lesions at 1 year. Here we present the outcomes of the same cohort of patients evaluated at 3 year follow-up. Micro-fragmented adipose tissue was obtained using a minimal manipulation technique in a closed system. The safety of the procedure was evaluated by recording type and incidence of any adverse event. The clinical outcomes were determined using the KOOS, IKDC-subjective, Tegner Lysholm Knee, and VAS pain scales taken pre-operatively and at 12 and 36 months follow-up. No adverse events, lipodystrophy cases at the harvesting site nor atypical inflammatory reactions at the joint level were reported. Of the 30 patients previously treated, one was lost, and seven received additional treatments in the period of observation. On average, the 22 patients that had no other treatments in the 3-year period showed that the results observed at 1 year were maintained. Moreover, 41, 55, 55 and 64% of the patients improved with respect to the 1-year follow-up in the Tegner Lysholm Knee, VAS, IKDC-subjective and total KOOS, respectively. Our results point to autologous and micro-fragmented adipose tissue injection as an innovative and safe approach for the management of diffuse degenerative knee chondropathy in the mid-term. The procedure is simple, affordable, minimally invasive, and compliant with the regulatory panorama.


Validation of a screening tool for the early identification of sepsis.
Sepsis is the leading cause of mortality in noncoronary intensive care units. Recent evidence based guidelines outline strategies for the management of sepsis and studies have shown that early implementation of these guidelines improves survival. We developed an extensive logic-based sepsis management protocol; however, we found that early recognition of sepsis was a major obstacle to protocol implementation. To improve this, we developed a three-step sepsis screening tool with escalating levels of decision making. We hypothesized that aggressive screening for sepsis would improve early recognition of sepsis and decrease sepsis-related mortality by insuring early appropriate interventions. Patients admitted to the surgical intensive care unit were screened twice daily by our nursing staff. The initial screen assesses the systemic inflammatory response syndrome parameters (heart rate, temperature, white blood cell count, and respiratory rate) and assigns a numeric score (0-4) for each. Patients with a score of > or = 4 screened positive proceed to the second step of the tool in which a midlevel provider attempts to identify the source of infection. If the patients screens positive for both systemic inflammatory response syndrome and an infection, the intensivist was notified to determine whether to implement our sepsis protocol. Over 5 months, 4,991 screens were completed on 920 patients. The prevalence of sepsis was 12.2%. The screening tool yielded a sensitivity of 96.5%, specificity of 96.7%, a positive predictive value of 80.2%, and a negative predictive value of 99.5%. In addition, sepsis-related mortality decreased from 35.1% to 23.3%. The three step sepsis screening tool is a valid tool for the early identification of sepsis. Implementation of this tool and our logic-based sepsis protocol has decreased sepsis-related mortality in our SICU by one third.


Trends in roll-your-own smoking: findings from the ITC Four-Country Survey (2002-2008).
To establish the trends in prevalence, and correlates, of roll-your-own (RYO) use in Canada, USA, UK and Australia, 2002-2008. Participants were 19,456 cigarette smokers interviewed during the longitudinal International Tobacco Control (ITC) Four-Country Survey in Canada, USA, UK, and Australia. "Predominant" RYO use (i.e., >50% of cigarettes smoked) increased significantly in the UK and USA as a proportion of all cigarette use (both P < .001) and in all countries as a proportion of any RYO use (all P < .010). Younger, financially stressed smokers are disproportionately contributing to "some" use (i.e., ≤50% of cigarettes smoked). Relative cost was the major reason given for using RYO, and predominant RYO use is consistently and significantly associated with low income. RYO market trends reflect the price advantages accruing to RYO (a product of favourable taxation regimes in some jurisdictions reinforced by the enhanced control over the amount of tobacco used), especially following the impact of the Global Financial Crisis; the availability of competing low-cost alternatives to RYO; accessibility of duty-free RYO tobacco; and tobacco industry niche marketing strategies. If policy makers want to ensure that the RYO option does not inhibit the fight to end the tobacco epidemic, especially amongst the disadvantaged, they need to reduce the price advantage, target additional health messages at (young) RYO users, and challenge niche marketing of RYO by the industry.


Consensus guidelines for the management of inflammatory bowel disease.
This is the first Brazilian Consensus on inflammatory bowel disease, carried out by the Brazilian Study Group of Inflammatory Bowel Disease, and discusses the treatment of Crohn's disease and ulcerative colitis in acute and remission phases. The first part of the text, brings out a review on the main drugs used in the treatment of inflammatory bowel disease, as well as their mechanisms of action and cautions during their use. In the second part, the committee's opinions about the most recommended medical and surgical approaches for both diseases are presented on the basis of disease activity, location and behaviour status. The recommendations here presented were widely discussed in several scientific meetings with active participation of all members of the group and were highly based on scientific evidence covered by the literature.


Uptake of various trace elements during germination of wheat, buckwheat and quinoa.
The practice of sprouting is widely used to improve the nutritional value of grain seeds. Several nutritive factors such as vitamin concentrations and bioavailability of trace elements and minerals increase during germination. The objective of this work was to study the enrichment of various essential trace elements during germination of wheat (Triticum aestivum), buckwheat (Fagopyrum esculentum), and quinoa (Chenopodium quinoa) seeds in order to improve their nutritional role as a source of bioavailable trace elements. Seeds were sprouted either in distilled- or tap-water and in five different electrolyte solutions to investigate the concentration-dependent uptake. The time-dependence was investigated by analyzing aliquots of the sprouts after certain germination periods. Samples were analyzed after freeze drying for their Li, V, Cr, Fe, Mn, Co, Cu, Zn, Sr, Mo, As and Se concentrations with inductively-coupled plasma mass-spectrometry (ICP-MS). As a control for possible changes in the biochemical metabolism of the sprouts, the biosynthesis of vitamin C was also determined by using reversed-phase ion-pair HPLC. It was shown that quinoa was the most resistant to the applied electrolyte solutions and had the highest uptake rates for almost all elements, followed by buckwheat and wheat. Greatest increases were observed for Co, Sr, and Li. No significant changes in vitamin C biosynthesis were observed between sprouts grown in different electrolyte solutions. The time-dependent uptake for most elements was characterized by a significant absorption during soaking of the seeds, followed by a lag phase during the first day of germination and an increased uptake during the second and third day. Se and As showed distinctly different uptake behaviors.


Phospholipid, cholesterol, polypeptide and glycoprotein composition of hepatic endosome subfractions.
The composition of hepatic endosome subfractions was compared directly with that of plasma membranes and Golgi-apparatus fractions. The neutral lipid and phospholipid composition of two endosome subfractions separated on Nycodenz gradients from a parent endosome fraction was similar to that of plasma membranes. The phospholipid/cholesterol ratios and the sphingomyelin contents were high, as in plasma membranes. However, the phosphatidylserine content was low. Endosomal subfractions contained a simpler polypeptide profile than did plasma membranes. However, a large number of glycoproteins were common to both fractions. Two endosome-specific glycoproteins, of Mr 59 000 and 38 000, were identified. Sialic acid was present at concentrations higher than in plasma membranes. The results indicate that endosomal membranes have a similar composition to plasma membranes that probably reflects their functional interaction during endocytosis and receptor recycling.


Evolution and Stagnation of Image Guidance for Surgery in the Lateral Skull: A Systematic Review 1989-2020.
Objective: Despite three decades of pre-clinical and clinical research into image guidance solutions as a more accurate and less invasive alternative for instrument and anatomy localization, translation into routine clinical practice for surgery in the lateral skull has not yet happened. The aim of this review is to identify challenges that need to be solved in order to provide image guidance solutions that are safe and beneficial for use during lateral skull surgery and to synthesize factors that facilitate the development of such solutions. Methods: Literature search was conducted via PubMed using terms relating to image guidance and the lateral skull. Data extraction included the following variables: image guidance error, imaging resolution, image guidance system, tracking technology, registration method, study endpoints, clinical target application, and publication year. A subsequent search of FDA 510(k) database for identified image guidance systems and extraction of the year of approval, intended use, and indications for use was performed. The study objectives and endpoints were subdivided in three time phases and summarized. Furthermore, it was analyzed which factors correlated with the image guidance error. Factor values for which an error ≤0.5 mm (μerror + 3σerror) was measured in more than one study were identified and inspected for time trends. Results: A descriptive statistics-based summary of study objectives and findings separated in three time intervals is provided. The literature provides qualitative and quantitative evidence that image guidance systems must provide an accuracy ≤0.5 mm (μerror + 3σerror) for their safe and beneficial application during surgery in the lateral skull. Spatial tracking accuracy and precision and medical image resolution both correlate with the image guidance accuracy, and all of them improved over the years. Tracking technology with accuracy ≤0.05 mm, computed tomography imaging with slice thickness ≤0.2 mm, and registration based on bone-anchored titanium fiducials are components that provide a sufficient setting for the development of sufficiently accurate image guidance. Conclusion: Image guidance systems must reliably provide an accuracy ≤0.5 mm (μerror + 3σerror) for their safe and beneficial use during surgery in the lateral skull. Advances in tracking and imaging technology contribute to the improvement of accuracy, eventually enabling the development and wide-scale adoption of image guidance solutions that can be used safely and beneficially during lateral skull surgery.


The Efficacy of Pralidoxime in the Treatment of Organophosphate Poisoning in Humans: A Systematic Review and Meta-analysis of Randomized Trials.
Introduction The benefits of atropine in the treatment of acute organophosphate (OP) poisoning has been well established, while that of oximes is still uncertain. Pralidoxime is the most often used oxime worldwide. In vitro experiments have consistently shown that oximes are effective reactivators of human acetylcholinesterase enzyme, inhibited by OP compounds. However, the clinical benefit of pralidoxime is still unclear. A recent meta-analysis has found that pralidoxime provides no significant improvement in outcome and rather may cause harm while increasing the economic burden in low-income communities where its use is the most prevalent. Objectives This study aimed to provide an updated evaluation of the efficacy of pralidoxime in addition to atropine alone in the treatment of patients with acute OP poisoning in terms of mortality, need for ventilator support, and the incidence of intermediate syndrome. The intermediate syndrome is a clinical syndrome that occurs 24 to 96 hours after the ingestion of an OP compound and is characterized by prominent weakness of neck flexors, muscles of respiration, and proximal limb muscles. Materials and methods We searched MEDLINE, EMBASE, CENTRAL, and ClinicalTrials.gov databases until January 2019 for randomized controlled trials (RCTs) in the English language that evaluated the use of pralidoxime in individuals of any age, gender or nationality presenting with an alleged history of OP intake. The primary outcome was mortality. Secondary outcomes were the need for ventilator support and the incidence of intermediate syndrome. The risk of bias in included studies was assessed using the tool recommended by the Cochrane Handbook of Systematic Review of Interventions. Treatment/control differences in these outcomes across included studies were combined using risk ratios (RR). Results Six randomized controlled trials (n = 646) fulfilled the inclusion criteria, including one further trial missed from the most recent systematic review. The risk of bias varied across studies, with Eddleston 2009 being of the lowest risk and Cherian 2005 being of high risk. The risk of mortality (RR = 1.53, 95% confidence interval (CI) 0.97 to 2.41, P = 0.07) and the need for ventilator support (RR = 1.29, 95% CI 0.97 to 1.71, P = 0.08) were not significantly different between the pralidoxime and the control group. There was a significant increase in the incidence of intermediate syndrome in the pralidoxime group (RR = 1.63; 95% CI 1.01 to 2.62, P = 0.04). Conclusions Based on our meta-analysis of the available RCTs, pralidoxime was not shown to be beneficial in patients with acute OP poisoning. Our findings are consistent with the other literature.


Comparison of pulsatile luteinizing hormone secretion between prepubertal children and young adults: evidence for a mass/amplitude-dependent difference without gender or day/night contrasts.
The assessment of pulsatile GnRH activity in children has become possible since the introduction of the sensitive third generation immunochemiluminescent assays, permitting detection of previously unmeasurable levels of LH and FSH. Despite this, however, studies differ with regard to pulse frequency and the presence of a diurnal variation in LH secretion in clinically prepubertal children. Discrepancies may reflect the limitations of relatively long intersampling intervals, less sensitive LH assays such as RIAs, and the use of algorithms to analyze pulsatile LH secretion, which do not account for endogenous production rates and metabolic clearance. To address this, we studied LH secretion in 10 prepubertal children (4 boys and 6 girls, age 8.5-10.8 y) and 12 young adults (7 men and 5 women in the early follicular phase, age 18.6-32.8 y). Blood was sampled every 5 min from 2000 h to 0200 h (nighttime) and from 0800 h to 1400 h (daytime) for LH determination, using an immunochemiluminescent assay. Deconvolution analysis revealed no difference between daytime and nighttime LH secretion, including LH secretory amplitude and pulse frequency, within any of the 4 groups, permitting pooling of the data from the 2 sampling intervals for analysis. In addition, there was no difference in LH secretion or half-life between genders. Comparison of pulsatile LH secretion between children and adults revealed a marked increase in the mass and amplitude of LH secreted per burst. LH secretory burst mass rose 9.5-fold in females, increasing the mean LH concentration by nearly 13-fold and the production rate by nearly 9-fold. The trend in males was similar although less remarkable, with a 4.2-fold rise in LH secretory burst mass from childhood to adulthood. No differences in pulse frequency, interburst interval, half-life, or approximate entropy were found between prepubertal children and adults. We conclude that the maturational change in LH secretion occurs via a highly specific mass/amplitude-dependent mechanism without significant gender or day/night differences.


Genetic mismatch affects the immunosuppressive properties of mesenchymal stem cells in vitro and their ability to influence the course of collagen-induced arthritis.
The immunological and homing properties of mesenchymal stem cells (MSCs) provide a potentially attractive treatment for arthritis. The objective of this study was to determine effects of genetic disparity on the immunosuppressive potential of MSCs in vitro and in vivo within collagen induced arthritis (CIA). The ability of DBA/1, FVB and BALB/c MSC preparations to impact the cytokine release profile of CD3/CD28 stimulated DBA/1 T cells was assessed in vitro. The effect of systemically delivered MSCs on the progression of CIA and cytokine production was assessed in vivo. All MSC preparations suppressed the release of TNFα and augmented the secretion of IL-4 and IL-10 by stimulated DBA/1 T-cells. However, assessment of the ratio of IFNγ to IL-4 production indicated that the more genetically distant BALB/c MSCs had significantly less immunosuppressive capacity. Systemic delivery of BALB/c MSC resulted in an exacerbation of CIA disease score in vivo and a higher erosive disease burden. This was not seen after treatment with syngeneic or partially mismatched MSCs. An increase in serum levels of IL-1β was observed up to 20 days post treatment with allogeneic MSCs. An initial elevation of IL-17 in these treatment groups persisted in those treated with fully mismatched BALB/c MSCs. Over the course of the study, there was a significant suppression of serum IL-17 levels in groups treated with syngeneic MSCs. These data demonstrate a significant difference in the immunosuppressive properties of syngeneic and allogeneic MSCs in vitro and in vivo, which needs to be appreciated when developing MSC based therapies for inflammatory arthritis.


Sex differences in body weight gains following amygdaloid lesions in rats.
Lesions of the most posterodorsal aspects of the amygdala resulted in equal weight gains (mean = 58 g) in male and female rats during a 22-day observation period. However, the absolute weight gains in the first 5 days after lesions were greater in females (+41.4 g) than in males (+18.8 g), as were the longer-term gains relative to their respective control groups. In a second study with female rats, it was found that amygdaloid lesions had little effect on the estrous cycle and that ovariectomy resulted in additional excessive weight gains in both rats with sham lesions and those with amygdaloid lesions. The weight gains produced by amygdaloid lesions and ovariectomy were additive. It is concluded that there is a sex difference in weight gains after amygdaloid lesions, but that the lesion-induced obesity is independent of estrogen levels. Similarities to lesions of the ventromedial hypothalamus are noted, and an amygdaloid-ventromedial hypothalamic pathway for the regulation of feeding behavior is proposed.


Predicting Persistent, Limited, and Delayed Problematic Cannabis Use in Early Adulthood: Findings From a Longitudinal Study.
To identify risk profiles associated with patterns of problematic cannabis use in early adulthood. Data came from 1,229 participants in the Great Smoky Mountains Study, a prospective 20-year cohort study from 1993 to 2015 that is representative of western North Carolina with yearly assessments conducted from ages 9 and 16 years, and assessments at ages 19, 21, 26, and 30 years. Patterns of problematic cannabis use (i.e., DSM-5 cannabis use disorder or daily use) in early adulthood included the following: nonproblematic use in late adolescence (ages 19-21) and early adulthood (ages 26-30); limited problematic use in late adolescence only; persistent problematic use in late adolescence and early adulthood; and delayed problematic use in early adulthood only. Multinominal logistic regression models examined pairwise associations between these patterns and risk factors in childhood/early adolescence (ages 9-16) and late adolescence (ages 19-21). Risk factors included psychiatric disorders (e.g., anxiety, depressive), other substance use (smoking, alcohol, illicit drugs), and challenging social factors (e.g., low socioeconomic status, family functioning, peers). Sex and race/ethnicity (white, African American, American Indian) interactions were tested. The persistent pattern (6.7% of sample) was characterized by more anxiety disorders across development and more DSM-5 CUD symptoms during late adolescence compared to the limited pattern (13.3%), which, in turn, had more childhood family instability and dysfunction. The delayed pattern (3.7%) was characterized by more externalizing disorders, maltreatment, and peer bullying in childhood compared to those in nonproblematic users. There were no significant interactions of sex or race/ethnicity. Problematic cannabis use patterns during early adulthood have distinctive risk profiles, which may be useful in tailoring targeted interventions.


Fibulin-5 initiates epithelial-mesenchymal transition (EMT) and enhances EMT induced by TGF-beta in mammary epithelial cells via a MMP-dependent mechanism.
Epithelial-mesenchymal transition (EMT) is a normal physiological process that regulates tissue development, remodeling and repair; however, aberrant EMT also elicits disease development in humans, including lung fibrosis, rheumatoid arthritis and cancer cell metastasis. Transforming growth factor-beta (TGF-beta) is a master regulator of EMT in normal mammary epithelial cells (MECs), wherein this pleiotropic cytokine also functions as a potent suppressor of mammary tumorigenesis. In contrast, malignant MECs typically evolve resistance to TGF-beta-mediated cytostasis and develop the ability to proliferate, invade and metastasize when stimulated by TGF-beta. It therefore stands to reason that establishing how TGF-beta promotes EMT may offer new insights into targeting the oncogenic activities of TGF-beta in human breast cancers. By monitoring alterations in the actin cytoskeleton and various markers of EMT, we show here that the TGF-beta gene target, fibulin-5 (FBLN5), initiates EMT and enhances that induced by TGF-beta. Whereas normal MECs contain few FBLN5 transcripts, those induced to undergo EMT by TGF-beta show significant upregulation of FBLN5 messenger RNA, suggesting that EMT and the dedifferentiation of MECs override the repression of FBLN5 expression in polarized MECs. We also show that FBLN5 stimulated matrix metalloproteinase expression and activity, leading to MEC invasion and EMT, to elevated Twist expression and to reduced E-cadherin expression. Finally, FBLN5 promoted anchorage-independent growth in normal and malignant MECs, as well as enhanced the growth of 4T1 tumors in mice. Taken together, these findings identify a novel EMT and tumor-promoting function for FBLN5 in developing and progressing breast cancers.


Blockade of angiotensin signaling improves myocardial function in hypercholesterolemia independent of changes in eicosanoid release.
In hypercholesterolemia in the presence or absence of atherosclerosis, cardiovascular dysfunction and altered signaling of angiotensin, nitric oxide, or prostanoids are closely related to enhanced oxidant stress. We analyzed the potentially beneficial effects of the specific angiotensin-converting enzyme inhibitor enalapril and the specific angiotensin receptor blocker losartan on cardiac performance, eicosanoid metabolism, and parameters of oxidant stress in hypercholesterolemic animals. Guinea pigs were fed a 1% cholesterol diet for 8 weeks (Chol) with or without equieffective doses of either enalapril (1.5 mg/kg/d; Ena) or losartan (3 mg/kg/d; Los). Hemodynamics were analyzed in Langendorff hearts. Detection of eicosanoids was by enzyme immunoassay. Estimation of plasma xanthine oxidase (XO) activity was determined by spectrophotometry. In hypercholesterolemic guinea pigs, enhanced oxidant stress (e.g., increased plasma XO activities) was associated with profound myocardial and coronary (e.g., endothelial) dysfunction. Both enalapril and losartan lowered plasma cholesterol levels slightly, but only the angiotensin receptor antagonist effectively suppressed the increased plasma XO activities (from 11.4 +/- 0.7 to 7.6 +/- 2.2 U/L), and at the same time decreased the augmented coronary flow (from 26.0 +/- 1.0 to 23.0 +/- 1.0 mL/min/g tissue) observed in hypercholesterolemic animals. Assessment of left ventricular pressure and contractility (e.g., dp/dtmax) as well as the diastolic relaxation parameter (tau) revealed substantial myocardial dysfunction (systolic and diastolic) in Chol that was more substantially (and comparably) improved during administration of losartan (Los) than during enalapril (Ena). Surprisingly, angiotensin signaling blockade by either antagonist further suppressed the diminished coronary dilator responses to bradykinin (BK; not significant for enalapril) or adenosine (Ado) was demonstrated in Chol Langendorff hearts [delta CPPBK/Ado: from 5.0 +/- 0.5/0.9 +/- 0.1 to 4.4 +/- 1.5/0.4 +/- 0.1 (Ena) or to 1.9 +/- 0.5/0.4 +/- 0.1 (Los) cm2 (area under the curve), respectively]. Finally, as expected from control studies using heart preparations from normocholesterolemic guinea pigs, enhanced cardiac release of eicosanoids, prostacyclin, and thromboxane in Chol (0.48 +/- 0.03 and 0.6 +/- 0.1 ng/min/g) was augmented even further by treatment with enalapril (Ena: 1.6 +/- 0.4 and 1.0 +/- 0.1 ng/min/g), but was significantly reduced to or below control levels in losartan-treated animals (Los: 0.4 +/- 0.1 and 0.2 +/- 0.1 ng/min/g). Blockade of angiotensin signaling via angiotensin-converting enzyme inhibition or receptor antagonism--although differentially acting on enhanced cardiac prostanoid metabolism and oxidant stress--efficiently restored proper systolic and diastolic myocardial performance (losartan was more beneficial than enalapril), probably by counterbalancing altered angiotensin II-->angiotensin receptor signaling in the cardiovascular system of hypercholesterolemic animals. Impaired coronary vasodilator capacity seems to be irreversible after 8 weeks of a high-cholesterol diet, as shown by the unexpected lack of a dilator effect with both enalapril and losartan.


Steroid Anti-Inflammatory Effects Did Not Improve Organ Quality in Brain-Dead Rats.
Effect of glucocorticoid administration on improving the outcomes of kidney and liver allografts has not been clearly elucidated. This study investigated the effect of prednisolone administration after onset of brain death (BD) on kidney and liver in a controlled rat model of BD. BD was induced in rats by inflating an epidurally placed balloon catheter. Animals were treated with saline or prednisolone (5, 12.5, or 22.5 mg/kg) one hour after the onset of BD. After 4 hours of BD, experiments were terminated and serum and tissues were collected. Tissue gene and protein expression were measured for markers of inflammation, apoptosis, and cellular stress response markers. Prednisolone caused a reduction of plasma levels of IL-6, while the tissue expression of IL-6, IL-1β, and MCP-1 in both kidney and liver were also reduced. Creatinine plasma levels, complement (C3) expression, HSP-70, HO-1, Bcl2/BAX ratio, and PMN influx did not significantly change in kidney nor liver. Plasma AST and LDH levels were increased in the prednisolone treated group. Our results demonstrate prednisolone can has an anti-inflammatory effect mediated through reducing serum circulating cytokines. However, this anti-inflammatory effect does not translate into improved kidney function and indeed was associated with increased liver injury markers.


Insulin resistance in non-obese subjects is associated with activation of the JNK pathway and impaired insulin signaling in skeletal muscle.
The pathogenesis of insulin resistance in the absence of obesity is unknown. In obesity, multiple stress kinases have been identified that impair the insulin signaling pathway via serine phosphorylation of key second messenger proteins. These stress kinases are activated through various mechanisms related to lipid oversupply locally in insulin target tissues and in various adipose depots. To explore whether specific stress kinases that have been implicated in the insulin resistance of obesity are potentially contributing to insulin resistance in non-obese individuals, twenty healthy, non-obese, normoglycemic subjects identified as insulin sensitive or resistant were studied. Vastus lateralis muscle biopsies obtained during euglycemic, hyperinsulinemic clamp were evaluated for insulin signaling and for activation of stress kinase pathways. Total and regional adipose stores and intramyocellular lipids (IMCL) were assessed by DXA, MRI and (1)H-MRS. In muscle of resistant subjects, phosphorylation of JNK was increased (1.36±0.23 vs. 0.78±0.10 OD units, P<0.05), while there was no evidence for activation of p38 MAPK or IKKβ. IRS-1 serine phosphorylation was increased (1.30±0.09 vs. 0.22±0.03 OD units, P<0.005) while insulin-stimulated tyrosine phosphorylation decreased (10.97±0.95 vs. 0.89±0.50 OD units, P<0.005). IMCL levels were twice as high in insulin resistant subjects (3.26±0.48 vs. 1.58±0.35% H(2)O peak, P<0.05), who also displayed increased total fat and abdominal fat when compared to insulin sensitive controls. This is the first report demonstrating that insulin resistance in non-obese, normoglycemic subjects is associated with activation of the JNK pathway related to increased IMCL and higher total body and abdominal adipose stores. While JNK activation is consistent with a primary impact of muscle lipid accumulation on metabolic stress, further work is necessary to determine the relative contributions of the various mediators of impaired insulin signaling in this population.


Non-invasive Imaging of Rice Roots in Non-compacted and Compacted Soil.
Roots are the prime organ for nutrient and water uptake and are therefore fundamental to the growth and development of plants. However, physical challenges of a heterogeneous environment and diverse edaphic stresses affect root growth in soil. Compacted soil is a serious global problem, causing inhibition of root elongation, which reduces surface area and impacts resource foraging. Visualisation and quantification of roots in soil is difficult due to this growth substrate's opaque nature; however, non-destructive imaging technologies are now becoming more widely available to plant and soil scientists working to address this challenge. We have recently developed an integrated approach, combining X-ray Computed Tomography (X-ray CT) and confocal microscopy to image roots grown in compacted soil conditions from a plant to a cellular scale. The method is suited to visualize cellular responses of root tips grown in both non-compacted and compacted soils. This protocol presents a fully integrated workflow, including soil column preparation, creation of compaction conditions, plant growth, imaging, and quantification of root adaptive responses at a cellular scale.


[Low-invasive interventions in abdominal surgery].
Low-invasive interventions have now become the usual practice of a surgical unit. Nowadays operations on abdominal and small pelvic organs can be made by using the Russian equipment. In all, more than half the patients with cholelithiasis and its complications, chronic appendicitis, inguinal hernias were operated on by employing video endoscopic equipment (Endomedium, Kazan) and miniaccess-intervention instruments (SAN, Yekaterinburg). The outcomes of surgical treatment are summarized in the paper. The benefits and drawbacks of low-invasive intervention procedures, the advantages of various operation in different groups of patients are dealt with. The feasibility of combined operations and treatment in patients with cholelithiasis is discussed.


Biopsychosocial Processes of Health and Disease During the COVID-19 Pandemic.
The biopsychosocial model provides a useful perspective for understanding the development and characteristics of the COVID-19 pandemic and its anticipated long-term consequences for society as well as individuals. This article provides a biopsychosocial perspective on the COVID pandemic and an editorial comment on the articles in this Special Issue of Psychosomatic Medicine. Based on analysis of the PubMed database, it is shown that the attention to psychological and social factors is 74% higher in COVID-19-related articles compared to all other health-related scientific articles published during the same time-period (between 1/1/2020 and 4/18/2021). Specifically, 18.6% of the ≈123,500 articles addressing COVID-19-related topics also included psychological or social factors in their content vs. 10.7% of articles that did not address COVID-19. The biopsychosocial model is relevant to understanding the interrelationships among risk factors and the multidimensional clinical and psychosocial COVID-19 outcomes. Clinical outcomes directly related to COVID-19 range from severe but rare events (mortality and intensive care treatment) to less severe common outcomes such as positive screening tests for COVID-19 with or without symptoms. In addition, psychosocial outcomes range in severity from frequently observed reduced psychological wellbeing to less common clinical mood and anxiety disorders and, in rare cases, suicidality. The COVID-19 pandemic is characterized by an unusually strong and short-term link between social factors and biological aspects of the disease, without mediating psychological factors. After a review of the articles presented in this Special Issue, this editorial concludes with suggestions for biopsychosocial models in research on COVID-19 and other large-scale health threats.


Uridine Administration Promotes Normalization of Heart Mitochondrial Function in Dystrophin-Deficient Mice and Decreases Tissue Fibrosis.
The work shows the effect of the metabolic modulator uridine on the functioning and ultrastructure of heart mitochondria in dystrophin-deficient mdx mice. Intraperitoneal administration of uridine (30 mg/kg/day for 28 days) improved K+ transport and increased its content in the heart mitochondria of mdx mice to the level of wild-type animals. This was accompanied by a significant decrease in the level of malondialdehyde and an increase in the number of mitochondria in the heart of mdx mice. At the same time, uridine did not affect the hyperfunctionality of mitochondria in mdx mice, which manifested in an increase in the calcium retention capacity. Nevertheless, we noted that uridine causes a significant decrease in the level of fibrosis in the heart of mdx mice, which attested to a positive effect of therapy.


Protein components of bacteriophages lambda and lambda virulent.
Parallel studies have been made of the protein coats of the temperate bacteriophage lambda and of a deletion mutant, lambda virulent. A new method for preparing ghosts of both phages by the action of Cu(++) is described. Protein ghosts of both phages can be dissolved in citrate at pH values below 3, more rapidly in the presence of 8 m urea. Both phages yielded three apparently identical protein components which can be separated by thin-layer gel filtration and thin-layer gel electrophoresis. The protein of molecular weight 47,000 +/- 1,500 represents about 55% of the protein of the ghosts and is therefore likely to be the subunit of the head. The other proteins of molecular weight 30,000 +/- 1,500 and 16,000 +/- 1,500 represent approximately 25% and 20% of the protein, respectively. Amino acid analyses of the ghosts from the two phages have been carried out and show no significant differences. The buoyant density of phage lambda virulent is 0.016 g/ml less than that of lambda. Since no differences have been found in the protein components of the two phages, this indicates that the virulent mutant contains approximately 16% less deoxyribonucleic acid than the temperate phage.


Incidence and predictors of post-ERCP pancreatitis in patients with suspected sphincter of Oddi dysfunction undergoing biliary or dual sphincterotomy: results from the EPISOD prospective multicenter randomized sham-controlled study.
Pancreatitis following endoscopic retrograde cholangiopancreatography (ERCP) is a significant and potentially life-threatening adverse event and is common in patients with suspected sphincter of Oddi dysfunction (SOD). Here we aimed to identify predictors of the risk in this population. The Evaluating Predictors and Interventions in SOD (EPISOD) study prospectively enrolled 214 post-cholecystectomy patients with SOD type III in seven US centers. Patients were randomized, using a 2:1 allocation, to sphincterotomy or sham procedure, irrespective of the results of sphincter of Oddi manometry. Patients in the sphincterotomy arm who had elevated pancreatic sphincter pressure were randomized to biliary only or to dual (biliary and pancreatic) sphincterotomy. All but one patient received prophylactic pancreatic stents, but none received pharmacological prophylaxis. Post ERCP pancreatitis (PEP) was defined as acute pancreatitis within the subsequent 7 days. Blinded research coordinators at each site called patients at 1 week post-procedure. PEP occurred in 26 patients, in 10.6 % (15/141) in the sphincterotomy arm and 15.1 % (11/73) in the sham arm; unadjusted relative risk 0.71 (95 % confidence interval [95 %CI] 0.34 - 1.46). PEP rate was not significantly different in patients who received sphincterotomy compared with those undergoing sham treatment. In addition, the proportion was not statistically different in those who received biliary sphincterotomy alone (12/94; 12.8 % [95 %CI 6.0 % - 19.5 %]) compared with dual sphincterotomy (3/47; 6.4 % [95 %CI 0.0 % - 13.4 %]). Multivariate analysis identified an interaction between duration of ERCP and sedation type (P < 0.02). The performance of biliary or dual sphincterotomy does not increase the risk of PEP in patients suspected of SOD. However, the high rate of PEP in patients with suspected SOD, despite pancreatic stenting in expert centers, is confirmed in this prospective study. The combined effect of duration of ERCP and sedation type on the development of PEP should be further explored.Clinicaltrials.gov registration: NCT00688662.


Psychostimulant augmentation during treatment with selective serotonin reuptake inhibitors in men with paraphilias and paraphilia-related disorders: a case series.
We describe an open trial of psychostimulants (primarily methylphenidate sustained release [SR]) added to selective serotonin reuptake inhibitors (SSRIs; primarily fluoxetine) during the course of pharmacologic treatment of men with paraphilias and paraphilia-related disorders (PRDs). Twenty-six men with paraphilias (N = 14) or PRDs (N = 12) were assessed for life-time mood disorders and attention-deficit/hyperactivity disorder (ADHD) as defined by DSM-IV. All men were assessed at baseline for total sexual outlet and average time per day associated with paraphilia/PRD sexual behaviors. The indications for the addition of a psychostimulant to a stable dose of SSRI included the retrospective diagnosis of ADHD with persistent adult symptoms despite pharmacotherapy with an SSRI (N = 17); residual paraphilia/PRD fantasies, urges, and activities despite SSRI pharmacotherapy (N = 16); the persistence or presence of residual depressive symptoms despite SSRI pharmacotherapy (N = 6); relapse or loss of SSRI efficacy during the treatment of sexual impulsivity disorders (N = 4); and treatment of SSRI-induced side effects (N = 4). SSRI pharmacotherapy (mean +/- SD duration = 8.8+/-11.1 months) had statistically significant effects in diminishing paraphilia/PRD-related total sexual outlet (p < .001) and average time/day spent in paraphilia/PRD sexual behavior (p < .001). Addition of methylphenidate SR (mean dose = 40 mg/day; mean +/- SD duration = 9.6+/-8.2 months) was associated with additional statistically significant effects on paraphilia/PRD-related total sexual outlet (p = .003) and average time per day (p = .04) in addition to improvement of putative residual ADHD and depressive symptoms. Methylphenidate SR can be cautiously and effectively combined with SSRI antidepressants to ameliorate paraphilias and paraphilia-related disorders for the indications listed above.


[Quantitative evaluation of the content of antibodies to hepatitis of A virus in serial immunoglobulin preparations].
A total of 156 samples of several lots of immune serum globulin (ISG) were tested for antibodies to hepatitis A virus (anti-HAV). In this study the blocking variant of the solid-phase enzymoimmunological method was used. The samples under test were the production of 12 manufacturing enterprises in the USSR and I enterprise in Czechoslovakia. All the samples under test were found to contain anti-HAV within the limits from 2500-5000 to 20000-40000. The samples produced by different manufacturing enterprises showing no essential differences in the content of anti-HAV. Most preparations (93.6%) contained antibodies at levels of 5000-20000. The ISG proposed as the international standard preparation had a titer of 8000-10000. No differences in the content of anti-HAV were detected in comparing ISG whose shelf life expired 4 years before the test with ISG whose shelf life was to expire 3 years later. The results obtained in this study are discussed in connection with the immunoprophylaxis of viral hepatitis A.


Isolation and characterization of a novel pyrene-degrading Bacillus vallismortis strain JY3A.
The PAHs-degrading bacterium strain JY3A was newly isolated from the polluted soil in the Jinan Oil Refinery Factory, Shandong Province of China. The isolate was identified as Bacillus vallismortis with respect to its 16S rDNA sequence, DNA-DNA relatedness and fatty acid profiles, as well as various physiological characteristics. The strain was Gram-positive, motile, endospore forming, aerobic, oxidase and catalase-positive. The cells were 0.8-1.0μm wide and 2.0-2.5μm long, single or in pairs and sometimes in chains. Bacillus vallismortis strain JY3A could utilize naphthalene, phenanthrene, anthracene, pyrene, fluorene, benzene, toluene, phenol, methanol, ethanol, Tween 80, cyclohexane or catechol as sole carbon source. The strain alone removed 90.5% of pyrene at an initial concentration of 150ppm in 15days in the presence of 0.5% (w/w) Tween 80. However, in co-culture with Phanerochaete chrysosporium, JY3A reduced the concentration of pyrene by nearly 55.4% after 7days of incubation.


Synchrotron grazing incidence X-ray scattering study of the morphological structures in thin films of a polymethacrylate diblock copolymer bearing POSS moieties.
The morphological structures in thin films of a diblock copolymer of methyl methacrylate and polyhedral oligomeric silsesquioxane (POSS) functionalized methacrylate (PMMA-b-PMAPOSS) with a volume ratio of 13/87 were investigated in detail by using synchrotron grazing incidence small and wide-angle X-ray scattering (GISAXS and GIWAXS). In addition, its thermal properties were studied. Thin films of this diblock copolymer were found to undergo phase-separation during solvent-annealing with carbon disulfide and post thermal annealing. To quantitatively analyze the scattering data, GISAXS and GIWAXS formulas were derived and applied. Our detailed analysis found that cylinders of PMMA blocks are induced to form in the diblock copolymer films by solvent-annealing and are hexagonally packed in the PMAPOSS matrix, in which the cylinders are oriented vertically with respect to the film plane. In the solvent-annealed films, both the PMMA cylinders and the PMAPOSS matrix are featureless, i.e., amorphous. However, the post thermal annealing process induces aggregation of the POSS moieties, which results in the formation of crystals with an orthorhombic lattice unit cell. These crystals were found to consist of PMAPOSS block chains in a helical conformation in which the molecular PMAPOSS cylinders are aligned in the film plane. The formation of these crystals is induced by the ordering ability of the POSS moieties. The crystals were found to melt above 190 degrees C during heating and subsequent cooling. In contrast, the hexagonally packed structure of the PMMA cylinders in the solvent-annealed and post thermally annealed films was found to be retained during the heating and the subsequent cooling. In addition, the scattering analysis provides detailed structural parameters. The 2D GISAXS and GIWAXS patterns were reconstructed from the determined structural parameters by using the derived scattering formulas, and found to be in good agreement with the experimental patterns. Moreover, a model for the structure of the films of the diblock copolymer is proposed.


Initiation of simian virus 40 DNA synthesis in vitro.
Simian virus 40 (SV40) T antigen can efficiently initiate SV40 origin-dependent DNA synthesis in crude extracts of HeLa cells. Therefore, initiation of SV40 DNA synthesis can be analyzed in detail. We present evidence that antibodies which neutralize proliferating cell nuclear antigen (PCNA) inhibit but do not abolish pulse-labeling of nascent DNA. The lengths of DNA products formed after a 5-s pulse in the absence and presence of anti-PCNA serum averaged 150 and 34 nucleotides, respectively. The small DNAs formed in the presence of anti-PCNA serum underwent little or no increase in size during further incubation periods. The addition of PCNA to reaction mixtures inhibited with anti-PCNA serum largely reversed the inhibitory effect of the antiserum. The small nascent DNAs formed in the presence or absence of anti-PCNA serum products arose from the replication of lagging strands. These results suggest that a PCNA-dependent elongation reaction participates in the synthesis of lagging strands as well as leading strands. We also present evidence that in crude extracts of HeLa cells, DNA synthesis generally does not initiate within the core origin. Initiation of DNA synthesis outside of a genetically defined origin region has not been previously described in a eukaryotic replication system but appears to be a common feature of initiation events in many prokaryotic organisms. Additional results presented indicate that in the absence of nucleoside triphosphates other than ATP, the preinitiation complex remains within or close to the SV40 origin.