Genetic polymorphism and natural selection of Duffy binding protein of Plasmodium vivax Myanmar isolates.
Plasmodium vivax Duffy binding protein (PvDBP) plays an essential role in erythrocyte invasion and a potential asexual blood stage vaccine candidate antigen against P. vivax. The polymorphic nature of PvDBP, particularly amino terminal cysteine-rich region (PvDBPII), represents a major impediment to the successful design of a protective vaccine against vivax malaria. In this study, the genetic polymorphism and natural selection at PvDBPII among Myanmar P. vivax isolates were analysed. Fifty-four P. vivax infected blood samples collected from patients in Myanmar were used. The region flanking PvDBPII was amplified by PCR, cloned into Escherichia coli, and sequenced. The polymorphic characters and natural selection of the region were analysed using the DnaSP and MEGA4 programs. Thirty-two point mutations (28 non-synonymous and four synonymous mutations) were identified in PvDBPII among the Myanmar P. vivax isolates. Sequence analyses revealed that 12 different PvDBPII haplotypes were identified in Myanmar P. vivax isolates and that the region has evolved under positive natural selection. High selective pressure preferentially acted on regions identified as B- and T-cell epitopes of PvDBPII. Recombination may also be played a role in the resulting genetic diversity of PvDBPII. PvDBPII of Myanmar P. vivax isolates displays a high level of genetic polymorphism and is under selective pressure. Myanmar P. vivax isolates share distinct types of PvDBPII alleles that are different from those of other geographical areas. These results will be useful for understanding the nature of the P. vivax population in Myanmar and for development of PvDBPII-based vaccine.


Developing evidence that is fit for purpose: a framework for payer and research dialogue.
Matching the supply and demand of evidence requires an understanding of when more evidence is needed, as well as the type of evidence that will meet this need. This article describes efforts to develop and refine a decision-making framework that considers payers' perspectives on the utility of evidence generated by different types of research methods, including real-world evidence. Conceptual framework development with subsequent testing during a roundtable dialogue. The framework development process included a literature scan to identify existing frameworks and relevant articles on payer decision making. The framework was refined during a stand-alone roundtable in December 2013 hosted by the research team, which included representatives from public and private payers, pharmacy benefit management, the life sciences industry, and researchers. The roundtable discussion also included an application of the framework to 3 case studies. Application of the framework to the clinical scenarios and the resulting discussion provided key insights into when new evidence is needed to inform payer decision making and what questions should be addressed. Payers are not necessarily seeking more evidence about treatment efficacy; rather, they are seeking more evidence for relevant end points that illustrate the differences between treatment alternatives that can justify the resources required to change practice. In addition, payers are interested in obtaining new evidence that goes beyond efficacy, with an emphasis on effectiveness, longer-term safety, and delivery system impact. We believe that our decision-making framework is a useful tool to increase dialogue between evidence generators and payers, while also allowing for greater efficiency in the research process.


Correction to: More than a bit of fun: the multiple outcomes of a bioblitz.
[This corrects the article DOI: 10.1093/biosci/biac100.].


Can tetracycline-induced fatty liver in pregnancy be attributed to choline deficiency?
Pregnant women, especially those in their third trimester, are particularly susceptible to tetracycline-induced fatty liver. Experimental evidence obtained in rats suggest that the precipitous fall in maternal liver choline concentration nearing the end of pregnancy could lead to a severe reduction in the lipotropic activity of the liver. It is hypothesised that the liver in this state will be at much greater risk of developing fatty degeneration, especially if it is exposed to hepatotoxic agents such as tetracycline.


Magnetic resonance imaging in the evaluation of suspected osteonecrosis of the knee.
Magnetic resonance imaging (MRI) was performed on 19 patients with suspected or proven osteonecrosis of the knee. The results were compared to radionuclide and plain radiographic studies when possible. The patients were grouped into one of three categories: patients with disease predisposing them to osteonecrosis (e.g., systemic lupus erythematosus (SLE), steroid use, and renal transplants), older patients without risk factors with acute onset of symptoms,and patients with knee pain months or years following trauma. In six patients with symptoms and predisposing diseases, MRI was abnormal in four cases, all of whom had bilateral abnormalities. In the ten older patients with classical symptoms, MRI was abnormal in seven, and bilateral abnormalities were present in three patients. The three patients with a history of antecedent trauma had normal MRI studies. Two patients with history and scintigraphic evidence of osteonecrosis had negative MRI scans. MRI may be of value in patients with suspected or proven osteonecrosis of the knee by demonstrating bilateral disease in patients with unilateral symptoms, showing the extent of involvement, and establishing the presence or absence of bone marrow changes in patients with positive bone scans and negative plain films.


Biliverdin reductase/bilirubin mediates the anti-apoptotic effect of hypoxia in pulmonary arterial smooth muscle cells through ERK1/2 pathway.
Inhibition of pulmonary arterial smooth muscle cell (PASMC) apoptosis induced by hypoxia plays an important role in pulmonary arterial remodeling leading to aggravate hypoxic pulmonary arterial hypertension. However, the mechanisms of hypoxia acting on PASMC apoptosis remain exclusive. Biliverdin reductase (BVR) has many essential biologic roles in physiological and pathological processes. Nevertheless, it is unclear whether the hypoxia-induced inhibition on PASMC apoptosis is mediated by BVR. In the present work, we found BVR majorly localized in PASMCs and was up-regulated in levels of protein and mRNA by hypoxia. Then we studied the contribution of BVR to anti-apoptotic response of hypoxia in PASMCs. Our results showed that siBVR, blocking generation of bilirubin, reversed the effect of hypoxia on enhancing cell survival and apoptotic protein (Bcl-2, procasepase-9, procasepase-3) expression, preventing nuclear shrinkage, DNA fragmentation and mitochondrial depolarization in starved PASMCs, which were recovered by exogenous bilirubin. Moreover, the inhibitory effect of bilirubin on PASMC apoptosis under hypoxic condition was blocked by the inhibitor of ERK1/2 pathway. Taken together, our data indicate that BVR contributes to the inhibitory process of hypoxia on PASMC apoptosis, which is mediated by bilirubin through ERK1/2 pathway.


A 90-day toxicology study of high-amylose transgenic rice grain in Sprague-Dawley rats.
A transgenic rice line (TRS) with high amylose level has been developed by antisense RNA inhibition of starch branching enzymes. Compositional analysis of TRS demonstrated that the content of resistant starch (RS) was significantly higher compared to conventional non-transgenic rice. High level of RS is an important raw material in food industry and has various physiological effects for human health. In order to provide the reliable theory basis for field release of TRS rice, we evaluated the potential health effects of long-term consumption of the TRS. The 90-day toxicology feeding experiment was conducted in Sprague-Dawley rats fed with diets containing 70% of either TRS rice flour, its near-isogenic rice flour or the control diet. The clinical performance variables (body weight, body weight gain and food consumption) were measured and pathological responses (hematological parameters and serum chemistry at the midterm and the completion of the experiment, urinalysis profile and serum sex hormone response at the completion of the experiment) were performed. Besides, clinical signs, relative organ weights and microscopic observations were also compared between TRS group and its near-isogenic rice group. The combined data indicates that high-amylose TRS grain is as safe as the conventional non-transgenic rice for rat consumption.


Keratinocyte-Macrophage Crosstalk by the Nrf2/Ccl2/EGF Signaling Axis Orchestrates Tissue Repair.
Unveiling the molecular mechanisms underlying tissue regeneration provides new opportunities to develop treatments for diabetic ulcers and other chronic skin lesions. Here, we show that Ccl2 secretion by epidermal keratinocytes is directly orchestrated by Nrf2, a prominent transcriptional regulator of tissue regeneration that is activated early after cutaneous injury. Through a unique feedback mechanism, we find that Ccl2 from epidermal keratinocytes not only drives chemotaxis of macrophages into the wound but also triggers macrophage expression of EGF, which in turn activates basal epidermal keratinocyte proliferation. Notably, we find dysfunctional activation of Nrf2 in epidermal keratinocytes of diabetic mice after wounding, which partly explains regenerative impairments associated with diabetes. These findings provide mechanistic insight into the critical relationship between keratinocyte and macrophage signaling during tissue repair, providing the basis for continued investigation of the therapeutic value of Nrf2.


The cost of a late-detected outbreak among people who inject drugs. A modeling study.
People who inject drugs (PWID) are at risk for human immunodeficiency virus (HIV) and hepatitis C virus (HCV). In 2009 and 2011, Athens, Greece experienced an HCV and an HIV outbreak among PWID, respectively. Of these, only the 2011 HIV outbreak was detected. However, the public health interventions implemented in response to the HIV outbreak tackled also indirectly the undetected HCV outbreak. The aim of this study is to highlight the potential benefits of an efficient notification system using as a case study the undetected 2009 HCV outbreak among PWID of Athens. More specifically, the study assesses whether an earlier implementation of the same public responses could diminish the scale of the HCV outbreak and estimates the potential cost-savings. A previous dynamic, stochastic, individual-based model was used to simulate HCV transmission among PWID of Athens, Greece. We calibrated the model to reproduce the observed HCV prevalence. We examined the effect of the non-detection scenario, the 1- or 2-years earlier detection scenarios and compared them to the status quo scenario. Under the non-detection scenario, 2800 additional PWID would have been infected with HCV compared to the status quo by 2019. On the contrary, if the outbreak was detected 1- or 2- years earlier with immediate interventions, 440 and 970 HCV cases could be averted by 2019, respectively. Non-detection of the outbreak would cost an additional 43.2 (95% Credible interval: 2.7, 59.4) million euros to the healthcare system, compared to the status quo. On the other hand, if there was an efficient notification system to detect the HCV outbreak 1 or 2 years earlier, 6.8-15.6 million euros could have been saved by 2019. An efficient notification system among PWID is a cost-saving investment that could detect on time and contain future outbreaks, and save valuable resources of the healthcare system.


Randomized trial examining effectiveness of lifestyle intervention in reducing gestational diabetes in high risk Chinese pregnant women in Hong Kong.
Gestational diabetes mellitus (GDM) is of public health concern. This trial examined whether a clinically proven lifestyle modification program (LMP) in early pregnancy was superior to routine antenatal care in improving GDM, maternal and infant outcomes. Chinese pregnant women at risk of GDM (n = 220) were recruited at or before 12-week gestation and randomized to either a LMP group or a routine care control group. Eighty subjects completed a dietitian-led LMP including dietary and exercise components from early pregnancy till 24-week gestation. Data were compared with those of 86 control subjects. Twenty three (26.7%) control subjects and 20 (25.0%) LMP subjects developed GDM (p = 0.798). The proportion of infants born large for gestational age and macrosomia was similar between groups. The LMP group showed a lower proportion of excessive gestational weight gain (GWG). Subgroup analysis suggested that those with higher LMP adherence showed more desirable dietary composition and energy intake, and lower proportion of excessive GWG compared with the low LMP adherence group and the control group. The potential effect of LMP on GDM and other maternal and infant outcomes, in particular GWG, as well as barriers for making lifestyle changes warrant further investigations (ClinicalTrials.gov NCT02368600).


Renal artery pseudoaneurysm after partial nephrectomy: three case reports and a literature review.
To report 3 cases of renal artery pseudoaneurysm (RAP) after partial nephrectomy and review the literature to discuss their unique presentation, clinical course, and management. RAPs are a rare clinical entity. They are a well-documented source of hemorrhage in association with renal biopsy, renal trauma, renal transplant, and percutaneous procedures; however, little is known about their presentation, pathophysiology, and management after nephron-sparing surgery. Between 2000 and 2002, 698 partial nephrectomies were performed at our institution for treatment of suspected localized renal malignancy. A retrospective review of these cases revealed that 3 patients developed a postoperative RAP. These cases were reviewed along with the current medical literature available from the MEDLINE database to characterize this clinical entity further. Three patients developed RAP after 698 uncomplicated partial nephrectomies were performed (0.43% incidence). Illustrating a classic presentation, the patient in case 1 was successfully treated with selective embolization after presenting with symptoms 3 weeks postoperatively. The patient in case 2 was treated with elective partial selective embolization for an asymptomatic lesion that later resolved spontaneously. In case 3, the patient presented early during the hospital course and required urgent selective embolization. RAP after nephron-sparing surgery is a rare, but potentially life-threatening, condition that is often difficult to diagnose and requires a high index of suspicion. Treatment should be tailored to each patient's clinical presentation to minimize morbidity and maximize renal conservation. All urologists should be aware of this potential postoperative complication and prepare accordingly.


Binding pose and affinity prediction in the 2016 D3R Grand Challenge 2 using the Wilma-SIE method.
The Farnesoid X receptor (FXR) exhibits significant backbone movement in response to the binding of various ligands and can be a challenge for pose prediction algorithms. As part of the D3R Grand Challenge 2, we tested Wilma-SIE, a rigid-protein docking method, on a set of 36 FXR ligands for which the crystal structures had originally been blinded. These ligands covered several classes of compounds. To overcome the rigid protein limitations of the method, we used an ensemble of publicly available structures for FXR from the PDB. The use of the ensemble allowed Wilma-SIE to predict poses with average and median RMSDs of 2.3 and 1.4 Å, respectively. It was quite clear, however, that had we used a single structure for the receptor the success rate would have been much lower. The most successful predictions were obtained on chemical classes for which one or more crystal structures of the receptor bound to a molecule of the same class was available. In the absence of a crystal structure for the class, observing a consensus binding mode for the ligands of the class using one or more receptor structures of other classes seemed to be indicative of a reasonable pose prediction. Affinity prediction proved to be more challenging with generally poor correlation with experimental IC50s (Kendall tau ~ 0.3). Even when the 36 crystal structures were used the accuracy of the predicted affinities was not appreciably improved. A possible cause of difficulty is the internal energy strain arising from conformational differences in the receptor across complexes, which may need to be properly estimated and incorporated into the SIE scoring function.


A modern and practical laccase-catalysed route suitable for the synthesis of 2-arylbenzimidazoles and 2-arylbenzothiazoles.
Heterocyclic aromatic compounds containing an imine (C[double bond, length as m-dash]N) bond such as benzimidazoles and benzothiazoles are important active pharmaceutical ingredients. The synthesis of 2-aryl-1H-benzimidazoles and 2-arylbenzothiazoles in good to excellent yields was achieved by reacting 2-aminoaromatics with various benzaldehyde derivatives catalysed by the commercial laccases Novoprime and Suberase® at room temperature and in the presence of atmospheric oxygen.


Chromoxane cyanine R. I. Physical and chemical properties of the dye and of some of its iron complexes.
Chromoxane cyanine R (Colour Index No. 43820, Mordant blue 3; also known as eriochrome cyanine R and solochrome cyanine R) is a valuable biological stain. The dyestuff is supplied as a powder containing a little less than 50% by weight of the monosodium salt of the dye, mixed with colourless crystalline and amorphous fillers. The tetrabasic colour acid was prepared and purified for study of the chemical and spectral properties of the dye. Chromoxane cyanine R is an acid-base indicator, with five different colours corresponding to the colour acid and the four anions. The most conspicuous colour change, from yellow to blue, occurs with ionization of the phenolic hydroxyl group at pH 11-12. The dye is assayed by measuring the absorbance of a strongly alkaline solution at 585 nm, with reference to a standard solution prepared from the purified colour acid. Spectrophotometric evidence has been found for the existence of three dye-metal complexes in solutions of the dye containing added ferric chloride at pH 1.5 (the pH of iron-dye solutions most useful in histological staining). These have the postulated compositions [ Fe2H (dye)]- (red), [ FeH2 (dye)]- (red), and [Fe2(dye)]2- (blue). The first two are probably simple carboxylate complexes of low stability. Increase in pH or molar iron: dye ratio promotes formation of the more stable blue complex, which is a metal chelate. Other blue complexes have been described by other investigators in solutions less acid than those that are useful in microtechnique. The production of blue and various shades of red in tissues stained by solutions containing iron (III) and chromoxane cyanine R probably involves reactions of both the red and the blue complexes of the dye.


It takes a community: The North Carolina Division of Public Health and the North Carolina area health education center program partner to reduce strokes and heart attacks.
The North Carolina Division of Public Health is leading a statewide project using a combination of approaches to address cardiovascular risk factors including obesity, hypertension, diabetes, and cigarette smoking. The objectives are to decrease tobacco use, increase physical activity, improve nutrition, and increase access to evidence-based clinical preventive services targeting hypertension, hypercholestrolemia, tobacco use, and weight management.


Prophylactic antiseizure drugs for spontaneous intracerebral hemorrhage: An updated systematic review and meta-analysis.
There is concern that recommendations on prophylactic antiseizure drugs (PASDs) for patients with spontaneous intracerebral hemorrhage (sICH) are biased by studies using older drugs and no electrographic monitoring. We performed a systematic review and meta-analysis to determine whether PASDs in patients with sICH reduced seizure occurrence and improved functional outcomes. We included analyses of newer trials, newer antiseizure drugs, and effectiveness in patients with consistent electrographic monitoring. Medline, Embase, and Cochrane were searched from inception until 12 August 2022, to identify studies with patients with sICH treated with PASDs, regardless of study design. The studied outcomes were functional status and occurrence of seizures. Fourteen studies were included, including 6742 patients. Risk of bias was low overall. There was no effect of PASD on seizure occurrence overall (odds ratio (OR) 0.73, 95% confidence interval (CI) 0.47-1.15), but they were associated with reduced occurrence in studies with electrographic monitoring (OR 0.36, 95% CI 0.18-0.70). There was no effect of PASDs on functional outcomes (OR 1.15; 95% CI 0.91-1.47) or mortality (OR 0.85, 95% CI 0.65-1.11). Prophylactic antiseizure medications after sICH reduce seizures in studies with electroencephalogram monitoring in high-risk patients. However, this benefit did not reflect in the improvement of functional outcomes, even in studies with newer, less toxic, antiseizure drugs.


Effect of acute hypoxic shock on the rat brain morphology and tripeptidyl peptidase I activity.
Hypoxic events are known to cause substantial damage to the hippocampus, cerebellum and striatum. The impact of hypoxic shock on other brain parts is not sufficiently studied. Recent studies show that tripeptidyl peptidase I (TPPI) activity in fish is altered after a hypoxic stress pointing out at a possible enzyme involvement in response to hypoxia. Similar studies are not performed in mammals. In this work, the effect of sodium nitrite-induced acute hypoxic shock on the rat brain was studied at different post-treatment periods. Morphological changes in cerebral cortex, cerebellum, medulla oblongata, thalamus, mesencephalon and pons were assessed using silver-copper impregnation for neurodegeneration. TPPI activity was biochemically assayed and localized by enzyme histochemistry. Although less vulnerable to oxidative stress, the studied brain areas showed different histopathological changes, such as neuronal loss and tissue vacuolization, dilatation of the smallest capillaries and impairment of neuronal processes. TPPI activity was strictly regulated following the hypoxic stress. It was found to increase 12-24h post-treatment, then decreased followed by a slow process of recovery. The enzyme histochemistry revealed a temporary enzyme deficiency in all types of neurons. These findings indicate a possible involvement of the enzyme in rat brain response to hypoxic stress.


Low-intensity climbing with blood flow restriction over 5 weeks increases grip and elbow flexor endurance in advanced climbers: A randomized controlled trial.
Grip and elbow flexor strength and endurance are crucial performance surrogates in competitive climbing. Thus, we examined the effects of blood flow restricted (BFR) climbing on grip and elbow flexor performance. Fifteen trained climbers (8 females; 20.8 ± 7.0 yrs; 1.72 ± 0.08 m; 63.0 ± 9.7 kg; 21.7 ± 2.7 IRCRCA grade) were either assigned to the intervention (BFR) or control (noBFR) group, using the minimization method (Strata: age, height, body mass, gender, and IRCRA grade). While BFR was used during low-intensity climbing training (2-times 10 min/session; 3-times/week), noBFR followed identical training protocols without BFR over 5 weeks. BFR of the upper limb was applied via customized pneumatic cuffs (occlusion pressure: 120 ± 23 mmHg, 75%; occlusion pressure). Endurance and strength performances were assessed via one-handed rung pulling (GripSTRENGTH), one-handed bent arm lock off at 90° (ArmSTRENGTH), static-intermitted finger hang (GripENDURANCE), and bent arm hang (ArmENDURANCE). Bayesian credible intervals revealed increased GripENDURANCE (+21 s (95% credible interval: -2 to 43 s)) and ArmENDURANCE +11 s (-5 to 27 s); adaptations via BFR. In contrast, GripSTRENGTH +4 N (-40 to 48 N) and ArmSTRENGTH +4 N (-68 to 75 N) were not affected by the BFR intervention. Fifteen cumulative sessions of BFR application with a cumulative total BFR load of 5 h over a 5 weeks macrocycle remarkably increased grip and elbow flexor endurance. Thus, BFR might serve as a promising means to improve relevant performance surrogates in trained climbers.


The food preservative ethoxyquin impairs zebrafish development, behavior and alters gene expression profile.
In the present study, we investigated the detrimental effects of ethoxyquin (EQ) on zebrafish embryonic development using different endpoints including lethality, malformations, locomotion and gene expression. EQ is primarily used as a preservative in animal feed and it has been shown to have negative impacts on different laboratory animals. However, studies on the adverse effects of EQ in aquatic animals are still limited. In this study, zebrafish eggs were exposed to different concentrations of EQ ranging from 1 to 100 μM for six days. In the 100 μM treated groups 95 and 100% mortality was observed at 24 and 48 h, respectively. Delayed development, decreased pigmentation and pericardial edema were observed in larvae. Behavioral analysis of larvae demonstrated a distinct locomotive pattern in response to EQ both in light and dark indicating a possible developmental neurotoxicity and deficits in locomotion. The expression levels of genes involved in several physiological pathways including stress response, cell cycle and DNA damage were altered by EQ. Our results demonstrate that EQ could cause developmental and physiological toxicity to aquatic organisms. Hence, its toxic effect should be further analyzed and its use and levels in the environment must be monitored carefully.


A sesquiterpene lactone glucoside from Ixeris denticulata f. pinnatipartita.
An extract from the entire Ixeris denticulata f. pinnatipartita plant afforded the guaianolide sesquiterpene lactone glucoside, 8 beta,15-dihydroxy-1(10),3,11(13)-guaiatrien-12,6-olide-15-O- glucopyranoside, along with the known flavonoids luteolin-7-O-glucoside and luteolin-7-O-glucuronide-6'-methyl ester; their structures were determined by spectroscopic methods. Ixerin Y inhibited the growth of human breast cancer MCF7 and MDA468 cell lines.


Information exchange between patients with Lynch syndrome and their genetic and non-genetic health professionals: whose responsibility?
Individuals at high risk for Lynch syndrome (LS) should be offered genetic counselling, since preventive options are available. However, uptake of genetic services and follow-up care are currently suboptimal, possibly caused by inadequate exchange of information. Therefore, this qualitative study aims to gain insight in the process of information exchange between patients diagnosed with LS and their non-genetic (i.e., general practitioner, gastroenterologist, gynaecologist) and genetic (i.e., clinical geneticist or genetic counsellor) health professionals concerning referral for genetic counselling and follow-up care. Participants comprised 13 patients diagnosed with LS (8 index patients and 5 of their affected relatives) and 24 health professionals (6 general practitioners, 8 gastroenterologists, 6 gynaecologists and 4 genetic health professionals). Analysis of the interview transcripts was performed in parallel and again after the interviews, following guidelines for qualitative research and using MAXQDA software. The main finding is that patients may 'get lost' between health professionals who lack a clear overview of their own and each other's role and responsibilities in the referral and follow-up care for patients with possible LS. Education of non-genetic health professionals and optimisation of communication between health professionals might help to enable more timely diagnosis of LS and allow patients to address their doubts and questions to the most appropriate healthcare professional.


Metabolic fate of combretastatin A-1: LC-DAD-MS/MS investigation and biological evaluation of its reactive metabolites.
Combretastatin A-1, an antineoplastic agent characterized by a remarkable cytotoxicity and a strong antitubulinic activity, is currently under investigation in phase I clinical trials. Yet a comprehensive metabolic study of CA-1 in human and animal models has so far not been reported in the literature. We have therefore investigated, through LC-UV and LC-MS analysis the in vitro/in vivo metabolism of CA-1, have synthesized its reactive quinone metabolite Q1, and have evaluated its cytotoxic and antitubulinic activities. In vitro CA-1 metabolism was studied in rat and human liver microsomes in the presence of the nucleophilic trapping agent GSH were performed while urine samples of the CA-1-treated rats were analyzed to establish the in vivo metabolic pathways. The metabolite Q1, that was synthesized in good yield using a polymer supported oxidant, displayed a significant cytotoxicity but was devoid of significant antitubulinic activity. Finally the chemical interaction of Q1 and the other combretastatin quinone metabolites with sulphydryl groups of tubulin was measured by Ellman's method: the results suggested the haptenization of the tubulin through the formation of a covalent bond.


Survival of 4 dogs with persistent atrial standstill treated by pacemaker implantation.
Pacemakers were implanted in 4 client-owned female dogs which had persistent atrial standstill. Three dogs were alive after 14 to 39 months and 1 dog was euthanized after 10.5 years. This report demonstrates that some dogs with persistent atrial standstill can survive for extended time periods.


Cytokine trafficking of IL-9 and IL-13 through TfnRc+ vesicles in activated human eosinophils.
Eosinophils are granulocytes that are elevated in lung mucosa in approximately half of patients with allergic asthma. These highly granulated cells can synthesize and secrete many cytokines, including IL-9 and IL-13. We hypothesized that IL-9 and IL-13 are found as preformed mediators in crystalloid granules and secreted using distinct trafficking pathways. Human eosinophils were purified from peripheral venous blood, adhered to coverslips, and stimulated with platelet activating factor (PAF). Cells were immunolabeled with antibodies to IL-9 or IL-13 and colocalized with markers for secretory organelles, using CD63 for crystalloid granules and transferrin receptor (TfnRc) for vesicles. Fixed cells were imaged using super-resolution microscopy and quantified by colocalization using Pearson's correlation coefficient. IL-9 immunofluorescence increased in a time-dependent manner to PAF, whereas colocalization of IL-9 and CD63 significantly increased from 0.52 to 0.67 after 5 min PAF. Colocalization of IL-9 with TfnRc significantly increased at 60 min of stimulation with PAF (0.54 at 0 min to 0.60 at 60 min). IL-13 showed lower colocalization with CD63 (0.55) than TfnRc (0.63) in unstimulated cells. Upon PAF stimulation, IL-13 intensity transiently decreased at 5 and 60 min, whereas colocalization of IL-13 with CD63 decreased throughout stimulation to 0.43. While colocalization of IL-13 with TfnRc transiently increased to 0.66 at 5 min PAF, it returned to near baseline levels (0.64) after 15 min PAF. Our results suggest that IL-9 and IL-13 are stored in crystalloid granules as well as endosomal structures, and that IL-9 is primarily trafficked to the cell surface via TfnRc+ endosome-like vesicles.


Aggressive osteoblastoma of the temporal bone: a case report and review of the literature.
Osteoblastoma can take place in any part of the skeleton. Its occurrence in the calvarium is extremely rare. The aggressive form of osteoblastoma is a rare variant. To our knowledge, this is the second case of aggressive osteoblastoma arising in the temporal bone. We report an 18-year-old patient with aggressive osteoblastoma involving the left temporal bone. Computed tomography of temporal bone showed an osteolytic lesion with granular mineralization and mottled ossified density. Permanent histopathologic sections and immunohistochemical stains revealed an aggressive osteoblastoma.


A new bisbenzylisoquinoline alkaloid isolated from Thalictrum foliolosum, as a potent inhibitor of DNA topoisomerase IB of Leishmania donovani.
Chemical investigation of the stem of Thalictrum foliolosum resulted in the isolation of two new bisbenzylisoquinoline alkaloids (1 and 2) along with known protoberberine group of isoquinoline alkaloids thalifendine (3) and berberine (4). The structures of the new compounds were established by detailed 2D NMR spectral analysis with their configurations determined from their optical rotation values and confirmed using circular dichroism. Inhibitory activities of these four compounds against DNA topoisomerase IB of Leishmania donovani were evaluated. Compound 2 exhibited almost complete inhibition of the enzyme activity at 50 μM concentration and it was found to be effective in killing both wild type as well as SAG resistant promastigotes of the parasite.


Thoracic splenosis.
Two rare cases of intrathoracic splenosis are reported. This phenomenon occurs almost exclusively in young men who have experienced splenic rupture and diaphragmatic laceration. The late occurrence of splenosis in the thoracic cavity poses problems in the differential diagnosis of solitary or multiple pleural-based nodules. It is important to recognize this entity from a clinical, radiographic, and therapeutic perspective.


Agricultural reuse of reclaimed water and uptake of organic compounds: pilot study at Mutah University wastewater treatment plant, Jordan.
The residues of polynuclear aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), chlorinated benzenes (CBs) and phenols were investigated for soil, wastewater, groundwater and plants. The uptake concentration of these compounds was comparatively determined using various plant types: Zea mays L., Helianthus annus L., Capsicum annum L., Abelmoschus esculentus L., Solanum melongena L. and Lycopersicon esculentum L. which were grown in a pilot site established at Mutah University wastewater treatment plant, Jordan. Soil, wastewater, groundwater and various plant parts (roots, leaves and fruits) samples were extracted in duplicate, cleaned up by open-column chromatography and analyzed by a multi-residue analytical methods using gas chromatography equipped with either mass selective detector (GC/MS), electron capture detector (GC/ECD), or flame ionization detector (FID). Environmentally relevant concentrations of targeted compounds were detected for wastewater much higher than for groundwater. The overall distribution profiles of PAHs and PCBs appeared similar for groundwater and wastewater indicating common potential pollution sources. The concentrations of PAHs, PCBs and phenols for different soils ranged from 169.34 to 673.20 microg kg(-1), 0.04 to 73.86 microg kg(-1) and 73.83 to 8724.42 microg kg(-1), respectively. However, much lower concentrations were detected for reference soil. CBs were detected in very low concentrations. Furthermore, it was found that different plants have different uptake and translocation behavior. As a consequence, there are some difficulties in evaluating the translocation of PAHs, CBs, PCBs and phenols from soil-roots-plant system. The uptake concentrations of various compounds from soil, in which plants grown, were dependent on plant variety and plant part, and they showed different uptake concentrations. Among the different plant parts, roots were found to be the most contaminated and fruits the least contaminated.


The proliferation of human tumor cell lines in the presence of different agars, agaroses, and methyl cellulose.
Human tumor cell lines, derived from cancers of the colon, ovary, and cervix, were grown in liquid tissue culture media and media made semisolid with agar (Bacto + deoxycholate lactose agar), agarose [LE, ME, Sea Plaque and Sea Prep (15/45)], and methyl cellulose. The effects of each agent on overall cell proliferation and rate of overall cell proliferation were examined. The agents, used to make media semisolid, were observed to inhibit or, in some cases, enhance cell growth in a fashion that was characteristic of individual cell lines. These phenomena may be of consequence to the optimization of nutrient media for primary tumor cell preparations.


Preventing Diabetes and Atherosclerosis in Sub-Saharan Africa: Should the Metabolic Syndrome Have a Role?
Obesity, hypertension, atherosclerosis, and type 2 diabetes mellitus are increasing in all regions of sub-Saharan Africa. The metabolic syndrome is a valuable tool in predicting atherosclerosis and type 2 diabetes in populations in Europe and North America. However, the applicability of the metabolic syndrome to African populations has not been studied. Prior to investing scarce funds into diagnosing and treating the metabolic syndrome, primary research needs to be designed to determine the prevalence of the metabolic syndrome and its ability to detect early, treatable disease in Africa. Assessment of these data should make it possible to determine if it is more effective to focus on the metabolic syndrome as a whole or on obesity, hypertension, atherosclerosis, and type 2 diabetes as individual conditions. This article is an overview of the presentation of metabolic syndrome variables in the Eastern, Western, and Southern regions of sub-Saharan Africa.


Treatment-induced plasticity in cerebral palsy: a diffusion tensor imaging study.
Diffusion tensor imaging is used as a measure of white-matter organization to probe mechanisms underlying clinical responses. Diffusion tensor imaging and clinical assessment in 8 patients with spastic quadriparesis (mean age, 6.13 years) was performed before and 6 months after therapy (botulinum injection, followed by physiotherapy). All patients were graded on the basis of gross motor function. Serial diffusion tensor imaging was also performed on 10 age/sex-matched controls at baseline and after 6 months. Regions of interests were placed on corticospinal tracts at different levels (i.e., corona radiata, posterior limb of internal capsule, midbrain, pons, and upper medulla) and on other major white-matter tracts, in both patients and controls. A significant increase in fractional anisotropy was evident in corticospinal tracts at the level of the posterior limb of the internal capsule and periventricular white matter of the temporal lobe, relative to baseline values in the patient group. Gross motor function classification system grades improved in all patients during follow-up relative to baseline values. The increase in fractional anisotropy in corticospinal tracts, along with improved clinical motor scores, suggests plasticity of the central motor pathway after combined therapy.


"M1/M2" Muscularis Macrophages Are Associated with Reduction of Interstitial Cells of Cajal and Glial Cells in Achalasia.
Several studies showed muscularis macrophages (MMφ) are associated with GI motility disorders. The purpose of this study was to preliminary explore the association between MMφ and achalasia. Tissue samples of the lower esophageal sphincter (LES) high-pressure zone were obtained from 27 achalasia patients and 10 controls. Immunohistochemistry for MMφ, interstitial cells of Cajal (ICC), neuronal nitric oxide synthase (nNOS), and glial cells were conducted. Histological characteristics were compared between groups, and correlation analysis was performed. Fewer ICC was found in achalasia compared with controls (P = 0.018), and the level of M1 macrophages was higher than that in controls no matter in terms of the number or the proportion of M1(P = 0.026 for M1 and 0.037 for M1/MMφ). Statistical differences were found between two groups in terms of proportion of M2 and ratio of M1 to M2 (P = 0.048 for M2/ MMφ and < 0.001 for M1/M2). For the correlation analysis, significant correlations were detected between levels of nNOS, ICC, and glial cells in patients with achalasia (P = 0.026 for nNOS and ICC, 0.001 for nNOS and glial cells, 0.019 for ICC and glial cells). There were significant correlations between M2/MMφ and levels of ICC (P = 0.019), glial cells (P = 0.004), and nNOS (P = 0.135). Patients with achalasia had a higher level of M1/M2 ratio in LES and significant correlations were found between M2/MMφ and numbers of ICC and glial cells, which suggested that MMφ were probably associated with occurrence and development of achalasia.


Street design scenarios using vegetation for sustainable thermal comfort in Erzurum, Turkey.
Urbanization models that do not comply with the planning criteria are affecting human lives. In urban areas, street trees have positive contributions to the ecosystem and human thermal comfort. In this study, the thermal comfort of the main streets that connect people to each other and provide access and transportation has been thermally explored. Cumhuriyet Street, which is one of the vibrant streets in Erzurum, was selected as a case study scenario in the winter and summer periods in 2018 by using the ENVI-met V. 4.4.2 winter model. A different green scenario is proposed, and the best thermal comfort scenario in both seasons is determined. The results show that, in the summer period, the air temperature of the greener street scenario is about 1.0 °C cooler than the existing condition and about 2.0 °C warmer in the winter period. Physiological equivalent temperature (PET) value was better in narrow canyon streets in winter months, but in wide canyon streets in summer months. The green scenarios of wide canyon streets positively affect the outdoor thermal comfort in both seasons. These results clearly imply that green streets are an appropriate strategy for city streets that suffer from discomfort levels in cold winter and hot summer periods. It has been concluded that it is possible to increase thermal comfort through improvement in the open space in street and more suitable plant preferences for livable urbanization. Planning streets in a new city characterized by summer and winter seasons should take into consideration an accurate decision for providing a thermal comfort level and healthy urbanization.


Lymphovascular space invasion does not predict vaginal relapses in stage I endometrioid adenocarcinoma of the endometrium.
This study was conducted to determine whether, in a pure population of patients with International Federation of Gynecology and Obstetrics stage I endometrioid endometrial (S1EE) carcinoma that is confined to the uterus and without lymph node metastases, the presence of lymphovascular space invasion (LVSI) is positively associated with vaginal relapses. Pathologic reports for all S1EE diagnosed in a hysterectomy specimen during a 9-year period (1997-2005) were reviewed. Cases with LVSI were selected and immunohistochemical staining for CD34, factor VIII-related antigen and pancytokeratin were performed on the relevant slides for confirmation. Various established prognostic/predictive clinicopathologic parameters were documented for the whole cohort and were correlated with the presence or absence of LVSI. One-way analysis of variance with Bonferroni correction and Fisher exact/chi(2) tests were used in the respective comparisons of continuous and categoric variables among the different groups. A total of 345 patients were diagnosed with S1EE during this period. The mean patient age for the cohort was 61.9 (+/-12.2) years (range, 28-89 years) and median follow-up was 80 months. Among these 345 patients, LVSI was present in 52 (15%), representing 1 (0.8%) of 121 stage 1A, 23 (14.5%) of 159 stage 1B, and 28 (43%) of 65 stage 1C cases (P < .001). Pelvic and/or para-aortic lymph node dissection was performed in 216 cases, and none had positive lymph nodes. We noted that LVSI correlated significantly with nuclear grade, with LVSI being present in 48% (10/21), 18% (32/175), and 5% (7/140) of cases with nuclear grades 3, 2, and 1 cases, respectively (P < .001). It was also more likely to be present in cases with architectural grades 2 and 3 (35/105) than grade 1 (16/234) (P < .01). Vaginal recurrences occurred in 2 (3.8%) of 52 patients with LVSI and 8 (2.7%) of 293 patients without LVSI (P = .65). In addition, on univariate analysis, LVSI did not correlate significantly with patient age, body mass index (using 50-year and 30 body mass index thresholds, respectively), or diabetic status. We conclude that LVSI is seen in approximately 15% of S1EE, and correlates with established prognostic parameters such as the level of myometrial invasion (substage), nuclear grade, and architectural grade. However, LVSI does not denote an increased likelihood of vaginal recurrences in S1EE. The presence of LVSI should not, in of itself, alter clinical staging, or trigger unnecessary therapeutic intervention.


Irritable Bowel Syndrome: Straightening the road from the Rome criteria.
A sequence of consensus-based Rome criteria for irritable bowel syndrome (IBS) has been published since 1989. The fundamental definition based on abdominal pain in association with bowel dysfunction has been consistent. However, two major changes occurred in the Rome II and IV criteria. The former change involved "splitting off" of symptoms that were not consistently associated with pain, such as functional, constipation, diarrhea, and bloating. In Rome IV, the main changes were the exclusion of discomfort (in contrast to pain) and the more stringent frequency criteria for the pain to be eligible for diagnosis of IBS (specifically, on average, at least 1 day per week in the last 3 months). Validation studies of the consensus, symptom-based criteria have identified multiple deficiencies that question the rationale for "splitting" the different syndromes, and favor a simpler identification of the classical symptoms of abdominal pain, bowel dysfunction, and bloating, and exclusion of alarm symptoms. Advances in the identification of actionable biomarkers related to the symptoms suggestive of functional gastrointestinal disorders have the potential to usher a change in practice from positive diagnosis of symptom complexes followed by empirical treatment to identification of the mechanisms causing the symptoms and targeted therapy.


How parents process child health and nutrition information: A grounded theory model.
The aim of the present study was to investigate low-income parents' experiences receiving, making meaning of, and applying sociocultural messages about childhood health and nutrition. Semi-structured interviews were conducted with parents from 16 low-income Early Head Start families. Verbatim interview transcripts, observations, field notes, documentary evidence, and follow-up participant checks were used during grounded theory analysis of the data. Data yielded a potential theoretical model of parental movement toward action involving (a) the culture and context influencing parents, (b) parents' sources of social and cultural messages, (c) parental values and engagement, (d) parental motivation for action, (e) intervening conditions impacting motivation and application, and (f) parent action taken on the individual and social levels. Parent characteristics greatly impacted the ways in which parents understood and applied health and nutrition information. Among other implications, it is recommended that educators and providers focus on a parent's beliefs, values, and cultural preferences regarding food and health behaviors as well as his/her personal/family definition of "health" when framing recommendations and developing interventions.


De novo transcriptome sequencing and analysis of salt-, alkali-, and drought-responsive genes in Sophora alopecuroides.
Salinity, alkalinity, and drought stress are the main abiotic stress factors affecting plant growth and development. Sophora alopecuroides L., a perennial leguminous herb in the genus Sophora, is a highly salt-tolerant sand-fixing pioneer species distributed mostly in Western Asia and northwestern China. Few studies have assessed responses to abiotic stress in S. alopecuroides. The transcriptome of the genes that confer stress-tolerance in this species has not previously been sequenced. Our objective was to sequence and analyze this transcriptome. Twelve cDNA libraries were constructed in triplicate from mRNA obtained from Sophora alopecuroides for the control and salt, alkali, and drought treatments. Using de novo assembly, 902,812 assembled unigenes were generated, with an average length of 294 bp. Based on similarity searches, 545,615 (60.43%) had at least one significant match in the Nr, Nt, Pfam, KOG/COG, Swiss-Prot, and GO databases. In addition, 1673 differentially expressed genes (DEGs) were obtained from the salt treatment, 8142 from the alkali treatment, and 17,479 from the drought treatment. A total of 11,936 transcription factor genes from 82 transcription factor families were functionally annotated under salt, alkali, and drought stress, these include MYB, bZIP, NAC and WRKY family members. DEGs were involved in the hormone signal transduction pathway, biosynthesis of secondary metabolites and antioxidant enzymes; this suggests that these pathways or processes may be involved in tolerance towards salt, alkali, and drought stress in S. alopecuroides. Our study first reported transcriptome reference sequence data in Sophora alopecuroides, a non-model plant without a reference genome. We determined digital expression profile and discovered a broad survey of unigenes associated with salt, alkali, and drought stress which provide genomic resources available for Sophora alopecuroides.


The effect of intranasal zinc sulfate treatment on odor-mediated behavior and on odor-induced metabolic activity in the olfactory bulbs of neonatal rats.
This study describes the effect of lesions of the peripheral olfactory receptor sheet on odor-mediated behavior and on odor-induced metabolic activity in the neonatal olfactory bulb. Nine-day-old rats were treated by intranasal irrigation with a solution of 1% ZnSO4 or 5% ZnSO4 or saline. At 1 and 5 days following treatment they were tested for maternally-directed behaviors which are mediated in large part by olfactory cues. At 1 day following treatment the pups treated with ZnSO4 solutions had significant deficits in their odor-directed behavior. By 5 days however, most pups treated with 1% ZnSO4 had recovered to control levels, while the pups treated with 5% ZnSO4 continued to exhibit profound behavioral deficits. Following the behavioral testing, the pups were tested using the 2-deoxyglucose method for amyl acetate odor-induced metabolic activity in their olfactory bulbs. Control pups had large regions of high focal activity in their anterolateral and caudomedial olfactory bulbs. The 1% ZnSO4 pups had smaller foci which were confined primarily to the medial olfactory bulb. The 5% ZnSO4 pups had very little focal uptake in their olfactory bulbs. There was a clear correlation between intact olfactory nerve and glomerular layers and sites of increased functional activity. The results of this study demonstrate that there is a relationship between the degree of behavioral deficit and the extent of odor-induced functional activity in the olfactory bulb.


Staining of endothelial cells does not change the result of cell density.
To determine the corneal endothelial cell density human donor corneas are usually placed in hypotonic balanced salt solution for unstained cell counting. Experimental studies often stain the corneal endothelium to highlight cells and cell borders and simplify counting. However, staining normally terminates the experiments. Up to date, there is no publication comparing endothelial cell counts before and after staining. This study compared the counting results of unstained (hypotonic balanced salt solution) and stained (trypan blue & alizarin red S) assessment of the corneal endothelial cell density. No evidence for significantly differing cell counts was found between unstained and stained analysis of the corneal endothelial cell density. Therefore, we consider unstained counting non-inferior to stained counting.


Linkage analysis merging replicate phenotypes: an application to three quantitative phenotypes in two African samples.
We report two approaches for linkage analysis of data consisting of replicate phenotypes. The first approach is specifically designed for the unusual (in human data) replicate structure of the Genetic Analysis Workshop 17 pedigree data. The second approach consists of a standard linkage analysis that, although not specifically tailored to data consisting of replicate genotypes, was envisioned as providing a sounding board against which our novel approach could be assessed. Both approaches are applied to the analysis of three quantitative phenotypes (Q1, Q2, and Q4) in two sets of African families. All analyses were carried out blind to the generating model (i.e., the "answers"). Using both methods, we found numerous significant linkage signals for Q1, although population colocalization was absent for most of these signals. The linkage analysis of Q2 and Q4 failed to reveal any strong linkage signals.


Improved protocols for the isolation and in-situ cryopreservation of cell colonies.
Stable transfection and cloning of cells often require physical separation of cell colonies. In order to conveniently isolate cell clones from petri dishes, we developed a protocol starting with a soft agar overlay of cells. This reduces the risk of cell diffusion between different colonies. Cells from individual colonies are mechanically removed, incubated with trypsin, and cell suspensions are seeded onto parallel microtiter plates. The cell clones on one microtiter plate can be cryopreserved in situ using the protocol described here which was tested for a variety of cell lines. Replica plates can be used for screening and further expansion of interesting clones. If screening can also be performed in situ, e.g., by immunocytochemistry, immunofluorescence, or the polymerase chain reaction, it is possible to perform most steps necessary in cell cloning experiments on microtiter plates.


Reduced work capacity after lumbar disc surgery: the role of cognitive-behavioral and work-related risk factors.
A significant number of patients who have had surgery for lumbosacral radicular syndrome still have a reduced work capacity several months later. In a prospective cohort study of 182 people who underwent lumbar disc surgery, we determined the predictive value of preoperatively measured cognitive-behavioral and work-related factors on work capacity 6 months after surgery. Logistic regression analyses indicated that these factors independently predicted work capacity 6 months after surgery. Specifically, fear of movement/(re)injury, more passive pain coping, and higher physical work-load predicted reduced work capacity in multiple logistic regression analyses, taking into account the role of a wide range of control variables including demographic variables, preoperative disability and pain intensity, neurological deficits, intake of analgesics, duration of complaints, and pain intensity 3 days postoperatively. The study supports the need to develop and evaluate preoperative risk screening measures that include both cognitive-behavioral and work-related factors and to evaluate the effectiveness of cognitive-behavioral and work-related interventions in patients at risk of reduced work capacity after surgery for LRS.


Use of lipoplex-induced nuclear factor-kappaB activation to enhance transgene expression by lipoplex in mouse lung.
Although lipofection-induced TNF-alpha can activate nuclear factor kappaB (NF-kappaB), which, in turn, increases the transgene expression from plasmid DNA in which any NF-kappaB responsive element is incorporated, no attempts have been made to use such biological responses as NF-kappaB activation against a vector to enhance vector-mediated gene transfer. A lipoplex composed of N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium and cholesterol liposome and plasmid DNA encoding firefly luciferase under the control of the cytomegalovirus immediate early promoter (pCMV-Luc) was intravenously injected into mice. Luciferase activity as well as NF-kappaB activation in the lung were evaluated. Then, a novel plasmid DNA, pCMV-kappaB-Luc, was constructed by inserting 5 repeats of NF-kappaB-binding sequences into the pCMV-Luc. NF-kappaB in the lung was activated by injection of the lipoplex and its nuclear localization was observed. An injection of lipopolysaccharide 30 min prior to the lipofection further activated NF-kappaB. At the same time, the treatment significantly increased the transgene expression by lipoplex, suggesting a positive correlation between expression and NF-kappaB activity. Based on these findings, we tried to enhance the lipoplex-based transgene expression by using NF-kappaB activation. The lipoplex consisting of pCMV-kappaB-Luc showed a 4.7-fold increase in transgene expression in the lung compared with that with pCMV-Luc. We demonstrated that NF-kappaB activation by lipoplex can be used to enhance lipoplex-mediated transgene expression by inserting NF-kappaB-binding sequences into plasmid DNA. These findings offer a novel method for designing a vector for gene transfer in conjunction with biological responses to it.


A new surgical drape support.
A lightweight, malleable surgical drape support attaches to the patient's forehead by sterile tape strips and rests comfortably over the patient's nose. It keeps the surgical drape from occluding the nasal and oral airway of the patient, allowing for comfort as well as observation during an ophthalmic surgical procedure performed under local anesthesia. It is sterilizeable and compact and thus does not impede the surgeon or assistant during microsurgical procedures.


Neuroradiological Features of Mild and Severe SARS-CoV-2 Infection.
An increasing number of neurological complications and corresponding radiological findings have been reported in patients with COVID-19 infection. The purpose of this study is to systematically review the current literature on COVID-19-associated neuroradiological findings and examine the prevalence of different findings in patients with both severe and mild COVID-19 infection. A comprehensive literature search of the PubMed and Embase databases was performed. Any studies reporting CT or MRI neuroimaging findings in patients with confirmed COVID-19 infection were included. Patient demographics, main radiological findings, neurological symptoms, and severity of COVID-19 infection were tabulated and quantified according to infection severity. Sixty-one studies published between 2019 and 2020 comprising 711 patients were analyzed according to severity of respiratory symptoms. The main neuroradiological findings for patients with mild classification were cranial nerve abnormalities, ischemic infarction, and white matter abnormalities, while the main findings in patients with severe classification were white matter abnormalities, ischemic infarction, and hemorrhagic events. Neuroradiological manifestations in COVID-19 infection are highly heterogeneous and differ based on the severity of COVID-19 infection. Cranial nerve abnormalities appear exclusive to mild infection, with a high degree of olfactory tract involvement, while hemorrhagic events are more common in severe infection. Notably, ischemic infarction was equally prevalent in both mild and severe COVID-19 infection. Healthcare providers treating COVID-19 patients should be aware of these potential complications and consider neurological assessment and neuroimaging studies when indicated.


Supralethal magnesemia with patient survival.
We report a patient with supralethal magnesemia of 38.7 mg/dL who survived. Our treatment consisted of assisted ventilation with intubation, accompanied by calcium chloride infusion and forced diuresis with mannitol.


Void formation by axial expansion of a gypsum-bonded investment.
The lengths of castings have been found to be dependent, in part, on the length of the ring lining material relative to the length of the casting ring. Five MOD inlay wax patterns were invested individually in open metal, in closed metal, and in plastic casting rings 35 mm or 42 mm long and 28 mm in diameter. The gingival walls of the wax patterns were placed either 8 mm or 15 mm from the uppermost ends of the rings. Specimens were broken in a mesiodistal direction, and the voids at the gingival walls were measured. The largest voids (139 microns) occurred in 42-mm-long plastic rings with the gingival walls 8 mm from the uppermost ends of the rings. The smallest voids (20 microns) occurred in the 35-mm-long closed metal rings. The boundaries between the investment and the voids were curved rather than accurately reflecting the straight gingival walls of the wax patterns.


[Cutaneous granulomatous reactions at botulinum neurotoxin A injection sites: First manifestation of systemic sarcoidosis].
We report a case of granulomatous skin reaction after injection of an unknown botulinum neurotoxin (BoNT) A preparation. Four years after occurrence of skin lesions, systemic sarcoidosis became manifest. In addition to injection trauma, the BoNT A preparation may have acted as an immunogenic stimulus leading to cutaneous manifestation of sarcoidosis.


Amelioration of inflammation and tissue damage in sickle cell model mice by Nrf2 activation.
Sickle cell disease (SCD) is an inherited disorder caused by a point mutation in the β-globin gene, leading to the production of abnormally shaped red blood cells. Sickle cells are prone to hemolysis and thereby release free heme into plasma, causing oxidative stress and inflammation that in turn result in damage to multiple organs. The transcription factor Nrf2 (nuclear factor erythroid 2-related factor 2) is a master regulator of the antioxidant cell-defense system. Here we show that constitutive Nrf2 activation by ablation of its negative regulator Keap1 (kelch-like ECH-associated protein 1) significantly improves symptoms in SCD model mice. SCD mice exhibit severe liver damage and lung inflammation associated with high expression levels of proinflammatory cytokines and adhesion molecules compared with normal mice. Importantly, these symptoms subsided after Nrf2 activation. Although hemolysis and stress erythropoiesis did not change substantially in the Nrf2-activated SCD mice, Nrf2 promoted the elimination of plasma heme released by sickle cells' hemolysis and thereby reduced oxidative stress and inflammation, demonstrating that Nrf2 activation reduces organ damage and segregates inflammation from prevention of hemolysis in SCD mice. Furthermore, administration of the Nrf2 inducer CDDO-Im (2-cyano-3, 12 dioxooleana-1, 9 diene-28-imidazolide) also relieved inflammation and organ failure in SCD mice. These results support the contention that Nrf2 induction may be an important means to protect organs from the pathophysiology of sickle cell-induced damage.


Evaluation of seven commercial RT-PCR kits for COVID-19 testing in pooled clinical specimens.
There are more than 350 real-time polymerase chain reaction (RT-PCR) coronavirus disease-2019 (COVID-19) testing kits commercially available but these kits have not been evaluated for pooled sample testing. Thus, this study was planned to compare and evaluate seven commercially available kits for pooled samples testing. Diagnostic accuracy of (1) TRUPCR SARS-CoV-2 Kit (Black Bio), (2) TaqPath RT-PCR COVID-19 Kit (Thermo Fisher Scientific), (3) Allplex 2019-nCOV Assay (Seegene), (4) Patho detect COVID-19 PCR kit (My Lab), (5) LabGun COVID-19 RT-PCR Kit (Lab Genomics, Korea), (6) Fosun COVID-19 RT-PCR detection kit (Fosun Ltd.), (7) Real-time Fluorescent RT-PCR kit for SARS CoV-2 (BGI) was evaluated on precharacterised 40 positive and 10 negative COVID-19 sample pools. All seven kits detected all sample pools with low Ct values (<30); while testing weak positive pooled samples with high Ct value (>30); the TRUPCR Kit, TaqPath Kit, Allplex Assay, and BGI RT-PCR kit showed 100% sensitivity, specificity, and accuracy. However, the Fosun kit, LabGun Kit, and Patho detect kit could detect only 90%, 85%, and 75% of weakly positive samples, respectively. We conclude that all seven commercially available RT-PCR kits included in this study can be used for routine molecular diagnosis of COVID-19. However, regarding performing pooled sample testing, it might be advisable to use those kits that performed best regarding positive identification in samples' pool, that is TRUPCR SARS-CoV-2 Kit, TaqPath RT-PCR COVID-19 Kit, Allplex 2019-nCOV Assay, and BGI Real-time RT-PCR kit for detecting SARS CoV-2.


Relationships between vitamin E status and erythrocyte stability in sheep.
Stability of erythrocytes from sheep with vitamin E deficiency (E-deficient) was compared by two haemolytic tests, one based on osmotic fragility in hypotonic saline and the other on detergent sensitivity in Tween 20. Relationships between haemolysis, plasma alpha-tocopherol concentration and creatine kinase (CK) activity were evaluated. E-deficient animals were clinically healthy but had elevated (greater than 1000 iu per 1) CK activities and low (less than 1 mumol per 1) alpha-tocopherol concentrations compared with control sheep (less than 500 iu per 1 and greater than 1.5 mumol per 1, respectively). Erythrocytes from E-deficient sheep were markedly more susceptible to detergent treatment than those from controls, but osmotic fragility was similar in both groups. Detergent sensitivity was directly related to plasma alpha-tocopherol concentration and CK activity but not to erythrocyte glutathione peroxidase activity. Within 24 h of supplementation (300 mg alpha-tocopherol subcutaneously) of E-deficient sheep, plasma alpha-tocopherol concentrations were increased, CK activities decreased and erythrocyte susceptibility to detergent-induced haemolysis was significantly reduced. Concentrations of alpha-tocopherol returned rapidly (less than 3 days) to pre-supplement values, although decreased haemolytic responses persisted longer (greater than 7 days). Cells from E-deficient animals were protected against detergent-induced haemolysis by preincubation with alpha-tocopherol in vitro. The results indicate that detergent sensitivity of erythrocytes may provide the basis of a simple functional test for vitamin E deficiency in sheep.


Eco synthesized chitosan/zinc oxide nanocomposites as the next generation of nano-delivery for antibacterial, antioxidant, antidiabetic potential, and chronic wound repair.
The present research work aimed at synthesizing chitosan-coated Zinc oxide nanocomposites (NS-CS/ZnONCs) by a bio-inspired method using an aqueous extract of Nigella sativa (NS) seeds and employing a quality-by-design approach (Box-Behnken design). The biosynthesized NS-CS/ZnONCs were physicochemically characterized and subjected to their in-vitro and in-vivo therapeutic potential. The zeta potential value of -11.2 mV and -12.6 mV indicated the stability of NS-mediated synthesized zinc oxide nanoparticles (NS-ZnONPs) and NS-CS/ZnONCs, respectively. The particle size of NS-ZnONPs and NS-CS/ZnONCs were 288.1 nm and 130.2 nm, respectively, with PDI of 0.198 and 0.158. NS-ZnONPs and NS-CS/ZnONCs showed superior radical scavenging abilities, excellent α-amylase, and α-glucosidase inhibitory activities. Also, NS-ZnONPs and NS-CS/ZnONCs demonstrated effective antibacterial activity against selected pathogens. Furthermore, NS-ZnONPs and NS-CS/ZnONCs demonstrated significant (p < 0.001) wound closure with 93.00 ± 0.43 % and 95.67 ± 0.43 % on the 15th day of treatment at the dose of 14 mg/wound, compared to 93.42 ± 0.58 % of standard. Collagen turnover was represented by hydroxyproline, which was shown to be significantly (p < 0.001) higher in the NS-ZnONPs (60.70 ± 1.44 mg/g of tissue) and NS-CS/ZnONCs (66.10 ± 1.23 mg/g of tissue) treatment groups than in the control group (47.7 ± 0.81 mg/g of tissue). Thus the NS-ZnONPs and NS-CS/ZnONCs could effectively develop promising drugs to inhibit pathogens and chronic tissue repair.


Zoledronic acid acutely increases sclerostin serum levels in women with postmenopausal osteoporosis.
Sclerostin is a circulating inhibitor of the Wnt-signaling pathway produced by osteocytes, which acts as a negative regulator of bone formation. Effects of zoledronic acid on sclerostin serum levels in postmenopausal osteoporosis are unknown. The purpose of this study was to evaluate sclerostin serum levels after zoledronic acid administration and correlate variations with bone turnover markers. We conducted a prospective intervention study in an ambulatory care setting. Forty women (mean age 62.6 ± 4.9 years) with postmenopausal osteoporosis were enrolled in this study and randomized into 2 groups to receive zoledronic acid (5 mg) or placebo. At baseline and then at 2, 7, 30, and 360 days after zoledronic acid or placebo administration, serum levels of sclerostin, bone-specific alkaline phosphatase (BSAP), as a bone formation marker, and serum C-telopeptide of type 1 collagen (CTX), as a bone resorption marker, were measured. Sclerostin serum levels increased by day 2, reached a peak at day 7 (3-fold baseline, P < .001), and then decreased at day 30 and returned near to baseline after 360 days in the zoledronic acid group. Both CTX and BSAP were reduced, and a significant negative correlation was observed between the percentage changes of sclerostin and the variation in BSAP and CTX at all time points in the zoledronic acid group (P < .05). No changes were observed in the placebo group. Our data demonstrate that zoledronic acid increases sclerostin serum levels and that sclerostin could play a role in coupling bone resorption to bone formation.


Effect of paper filter windows on indoor exposure to particles of outdoor origin.
Buildings are often located near ambient air pollution sources such as wildfire or heavy traffic areas. While windows in buildings are intermittently open for free cooling or natural ventilation, increased leakage area can lead to elevated human exposure to air pollutants of outdoor origin. The objective of this study is to investigate the effectiveness of paper filter windows in reducing exposure to outdoor air pollution and improving indoor air quality. The physical properties of paper windows as filtration media were experimentally determined, based on which multi-zone indoor air quality and ventilation analysis (CONTAM) simulations were performed for a full-scale building. The results show that the outdoor-indoor air exchange rate of a building can increase about 100% when conventional windows are replaced with paper filter windows. Even with the increased air exchange rate, the infiltration of outdoor particles into the building was reduced about 57-77% for the particle size range of 7-300 nm. These findings imply that paper windows have potential benefits for controlling both outdoor originated pollutants and indoor-generated pollutants with minimal energy inputs, especially in cities and communities impacted by urban air pollution and wildfires.


Effects of orchidectomy and male hormone administration on the adrenal of common myna, Acridotheres tristis.
When orchidectomized, the adrenals of semi-domestic, adult common myna, Acridotheres tritis atrophied. The cortical cord width and cortico-medullary ratio decreased and ascorbic acid and cholesterol contents of the adrenal increased significantly. Testosterone, depending upon the dose employed either restored the atrophic adrenals to normal condition or caused further atrophy of the glands of the castrated birds, suggesting a stimulatory effect on the adrenals at low and inhibitory at high dose levels.


Behavior of the muscle quality index and isometric strength in elderly women.
This study aimed (I) to compare the muscle quality index (MQI) and the isometric hip strength between younger and older women, and (II) to determine the relationship of the MQI with the sit-to-stand test (STS) and isometric hip strength in younger and older women. Twenty-eight elderly women (age= 66.2 ± 5.6 years) and twenty younger women (21.2 ± 2.2 years) participated in the study. The following dependent variables were measured: MQI, STS, body composition, and the peak isometric strength of the hip (PF) which was also normalized using three different methods (PF/Body Mass, PF/Fat-Free Mass, and PF/Body Mass0,335). Older women presented a lower PF in hip flexion, extension and external rotation regardless of the method of normalization (p < 0.001), but the PF of hip abductors and internal rotators was higher for older women (p < 0.05). No significant differences were found in the MQI between older and younger women (p = 0.443). The MQI was negatively correlated with the time in the STS in older women (r = -0.706, p < 0.001) and younger women (r = -0.729, p < 0.001), while the correlations of MQI with isometric hip strength were weaker in older women (r range: -0.082 - 0.556) and younger women (r range: -0.020 - 0.309). MQI is a clinical and practical tool to assess the muscular power of the lower extremities.


Multiplex liquid chromatography-tandem mass spectrometry for the detection of wheat, oat, barley and rye prolamins towards the assessment of gluten-free product safety.
Celiac patients should feel confident in the safety of foods labelled or expected to be gluten-free. In this context, a targeted proteomic approach based on liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) technique was proposed to assess the presence of celiotoxic cereals, namely wheat, oats, barley and rye, in raw and processed food products. To this aim, unique marker peptides were properly selected in order to distinguish between the different cereal types. A revised cocktail solution based on reducing and denaturing agents was exploited for prolamin extraction from raw and processed food; in addition, defatting with hexane was carried out for sample clean-up, allowing to largely reduce problems related to matrix effect. Method validation on fortified rice flour showed good analytical performance in terms of sensitivity (limits of detection in the 2-18 mg kg(-1) range). However, poor trueness was calculated for self-made incurred bread (between 3 and 30% depending on the peptide), probably due to baking processes, which reduce gluten extractability. Thus, it is evident that in the case of processed foods further insights into sample treatment efficiency and reference materials for protein calibration are required to obtain accurate gluten determination. Finally, the developed method was applied for the analysis of market food products, offering the possibility to discriminate among cereals, with good agreement with labelled ingredients for gluten-containing foodstuffs.


A deep neural framework for real-time vehicular accident detection based on motion temporal templates.
Vehicular accident prediction and detection has recently garnered curiosity and large amounts of attention in machine learning applications and related areas, due to its peculiar and fascinating application potentials in the development of Intelligent Transportation Systems (ITS) that play a pivotal role in the success of emerging smart cities. In this paper, we present a new vision-based framework for real-time vehicular accident prediction and detection, based on motion temporal templates and fuzzy time-slicing. The presented framework proceeds in a stepwise fashion, starting with automatically detecting moving objects (i.e., on-road vehicles or roadside pedestrians), followed by dynamically keep tracking of the detected moving objects based on temporal templates, clustering and supervised learning. Then, an extensive set of local features is extracted from the temporal templates of moving objects. Finally, an effective deep neural network (DNN) model is trained on the extracted features to detect abnormal vehicle behavioral patterns and thus predict an accident just before it occurs. The experiments on real-world vehicular accident videos demonstrate that the framework can yield mostly promising results by achieving a hit rate of 98.5% with a false alarm rate of 4.2% that compare very favorably to those from existing approaches, while still being able to deliver delay guarantees for realtime traffic monitoring and surveillance applications.


Infectious causes of pityriasis lichenoides: a case of fulminant infectious mononucleosis.
Pityriasis lichenoides is a rare cutaneous eruption of unknown cause that spans a spectrum of clinical severity. Infectious agents have long been suspected as etiologic factors. The present case is the first to demonstrate a known EBV-mediated process evolving and resolving in concert with pityriasis lichenoides. Epstein-Barr virus, Toxoplasma gondii, and HIV are the most frequently reported infectious triggers of pityriasis lichenoides. Pityriasis lichenoides may arise secondary to EBV-mediated acute infectious mononucleosis.


Distribution of isoaccepting tRNAs and codons for proline and glycine in collagenous and noncollagenous chicken tissues.
The relation between codon usage and tRNA content for proline and glycine, the major constituents of collagen, was studied in two tissues: the magnum of laying hen oviduct and the leg tendons of chick embryo where collagen is produced. Although the relative contents of tRNA(GCCGly) and tRNA(IGGPro) in tendons, as compared to magnum indicate a specialization of the tRNA population for collagen synthesis, the distribution of the preponderant codons in collagen mRNA is correlated but at a lesser extent to that of their cognate tRNAs.


[Rat liver S-adenosyl-L-homocysteine hydrolase purification by affinity column chromatography (author's transl)].
S-adenosyl-L-homocysteine hydrolase (EC 18.104.22.168) has been purified 240-fold from rat liver by affinity column chromatography on aminohexyl sepharose bound 6-mercaptopurine 9 D-riboside. The purified enzyme was homogeneous by gel electrophoresis.


Ventilator-associated pneumonia in a tertiary care ICU: analysis of risk factors for acquisition and mortality.
To investigate the incidence, risk factors and mortality of ventilator-associated pneumonia (VAP) in intensive care unit (ICU) patients. Prospective, observational, population-based study. The medical (14-bed) and surgical ICU (26-bed) of the Ghent University Hospital. All 1295 patients admitted to the ICU during 4 three-month periods between 1996 and 1998 were included. A set of demographic and clinical variables were collected at the day of admission and during the ICU course. The incidence of VAP among ICU patients ventilated at least 48 hours was 23.1%. The mean time to the development of VAP was 9.6 days with a median of 6 days. In the population of patients ventilated for at least 48 hours, a comparison was made between patients with (n = 89) and without VAP (n = 296). Patients with VAP had a significant longer ICU stay, with a longer ventilation dependency. Logistic regression analysis identified admission diagnosis other than trauma (OR: 0.51, 95% CI: 0.29-0.89; p = 0.02) and the length of ICU stay (OR: 1.05, 95% CI: 1.03-1.07; p < 0.001) to be independently associated with the acquisiton of VAP. In comparison with the total study population, patients with VAP had a higher ICU mortality (20.2% vs. 12.0%; p = 0.04), but not in the cohort group of patients at risk for VAP (ventilated > 48 hours)(20.2% vs. 31.3%; p = 0.03). The factors independently associated with death were higher SAPS II scores (OR 1.02, 95% CI: 1.003-1.032; p = 0.02), an admission diagnosis other than trauma (OR 0.36, 95% CI: 0.17-0.75; p = 0.006) and length of ICU stay (OR 0.97, 95% CI: 0.946-0.995; p = 0.02). This model did not recognize VAP as an independent predictor of death (OR 0.79, 95% CI: 0.41-1.53; p = 0.492). The incidence of VAP in our ICU is 23.1%. Length of ICU stay and an admission diagnosis other than trauma are major risk factors for the development of this nosocomial infection. VAP is associated with a high fatality rate. However, after adjustment for disease severity and length of ICU stay, VAP was not identified as an independent predictor of death.


Forerunners of mesh grafting machines. From cupping glasses and scarificators to modern mesh graft instruments.
Lanz described mesh grafting in 1907. A similar device was used earlier by Paré and Heister for different applications. Historical analysis reveals a definite link between old scarificators and mesh graft machines.


Backward bifurcations and multiple equilibria in epidemic models with structured immunity.
Many disease pathogens stimulate immunity in their hosts, which then wanes over time. To better understand the impact of this immunity on epidemiological dynamics, we propose an epidemic model structured according to immunity level that can be applied in many different settings. Under biologically realistic hypotheses, we find that immunity alone never creates a backward bifurcation of the disease-free steady state. This does not rule out the possibility of multiple stable equilibria, but we provide two sufficient conditions for the uniqueness of the endemic equilibrium, and show that these conditions ensure uniqueness in several common special cases. Our results indicate that the within-host dynamics of immunity can, in principle, have important consequences for population-level dynamics, but also suggest that this would require strong non-monotone effects in the immune response to infection. Neutralizing antibody titer data for measles are used to demonstrate the biological application of our theory.


Ogt-mediated O-GlcNAcylation inhibits astrocytes activation through modulating NF-κB signaling pathway.
Previous studies have shown that Ogt-mediated O-GlcNAcylation is essential for neuronal development and function. However, the function of O-GlcNAc transferase (Ogt) and O-GlcNAcylation in astrocytes remains largely unknown. Here we show that Ogt deficiency induces inflammatory activation of astrocytes in vivo and in vitro, and impairs cognitive function of mice. The restoration of O-GlcNAcylation via GlcNAc supplementation inhibits the activation of astrocytes, inflammation and improves the impaired cognitive function of Ogt deficient mice. Mechanistically, Ogt interacts with NF-κB p65 and catalyzes the O-GlcNAcylation of NF-κB p65 in astrocytes. Ogt deficiency induces the activation of NF-κB signaling pathway by promoting Gsk3β binding. Moreover, Ogt depletion induces the activation of astrocytes derived from human induced pluripotent stem cells. The restoration of O-GlcNAcylation inhibits the activation of astrocytes, inflammation and reduces Aβ plaque of AD mice in vitro and in vivo. Collectively, our study reveals a critical function of Ogt-mediated O-GlcNAcylation in astrocytes through regulating NF-κB signaling pathway.


Effects of obesity and weight-loss surgery shift the microbiome and impact alloimmune responses.
Obesity is a worldwide health problem with increasing rates in both children and adults. Bariatric surgery (BS) represents the only effective long-term treatment. Beneficial effects of BS may be mediated through shifts of the gut microbiome. Here, we introduce data linking the microbiome to alloimmune responses. The rapid development of microbiome sequencing technologies in addition to the availability of gnotobiotic facilities have enabled mechanistic investigations on modulations of alloimmune responses through microbiomes. BS has been shown to improve comorbidities and chronic inflammation caused by obesity. Changes in microbiota and microbiota-related metabolites may play a role. Patients either listed or having received a transplant have undergone weight loss surgery, thus allowing to dissect mechanisms of microbial shifts to alloimmunity. Weight loss and BS have the potential to improve transplant outcomes by ameliorating alloimmune responses. Those effects may be carried out through alterations of the gut microbiome.


TBHP-mediated oxidative synthesis of substituted pyrimido[4,5-d]pyrimidines from N-uracil amidines and methylarenes under metal free conditions.
An efficient and operationally simple protocol has been demonstrated for the synthesis of 1,3,5,7-tetrasubstituted pyrimido[4,5-d]pyrimidines via TBHP-mediated direct oxidative coupling of N-uracil amidines and methylarenes under metal-free conditions. Due to the inherent stability of methylarenes compared to aldehydes, the presented synthetic protocol is adaptable to a broad substrate scope, is operationally simple, has no need for stringent protection in the whole preparation process, and has the potential to prepare valuable products that are currently inaccessible or challenging to prepare using conventional methods. It is a significantly important complement to the conventional synthetic methods. The reaction possesses an efficient tandem oxidation-imination-cyclization process.


Development of the rat thalamus: IV. The intermediate lobule of the thalamic neuroepithelium, and the time and site of origin and settling pattern of neurons of the ventral nuclear complex.
Short-survival, sequential, and long-survival thymidine radiograms of rat embryos, fetuses, and young pups were analyzed in order to examine the time of origin, settling pattern, migratory route, and site of origin of neurons of the ventral nuclear complex of the thalamus. Quantitative examination of long-survival radiograms established that the bulk of the neurons of the ventral nuclear complex are generated between days E14 and E16 but with statistically significant differences between its three nuclei. The ventrobasal nucleus is the oldest component (97% of the cells are generated on days E14 and E15); the ventrolateral nucleus is next (82% of the cells are generated on days E14 and E15); and the ventromedial nucleus is last (51% of the cells are generated on days E14 and E15). In addition to this caudal-to-rostral (from the ventrobasal nucleus to the ventrolateral nucleus) and lateral-to-medial (from the ventrobasal nucleus to the ventromedial nucleus) internuclear gradients, there are lateral-to-medial and ventral-to-dorsal intranuclear neurogenetic gradients within the ventrobasal and ventrolateral nuclei. Qualitative examination of short and sequential survival thymidine radiograms indicate that the neurons of the ventral nuclear complex originate in the unique intermediate thalamic neuroepithelial lobule, which is distinguished from the rest of the thalamic neuroepithelium by the presence of a mitotically active secondary neuroepithelial matrix. Two sublobules can be distinguished in the intermediate lobule during the early stages of thalamic development. On the basis of their location and chronological pattern of cell production and differentiation, it is inferred that the neurons of the ventrobasal nucleus originate in the earlier differentiating, posteroventrally situated inverted sublobule, and the neurons of the ventrolateral nucleus are produced in the later differentiating, anterodorsally situated everted sublobule. The neurons of the ventromedial nucleus appear to originate from the intermediate neuroepithelial lobule after its two sublobules are no longer distinguishable. The heavily labeled neurons generated soon after injection on day E15 form a wave front that translocates in a lateral direction at a steady rate of 215 microns/day. Examination of methacrylate-embedded materials showed that, in day E15 rats the actively migrating cells are spindle-shaped, with their long axis oriented horizontally. The far-laterally situated differentiating cells (the oldest neurons) become vertically oriented by day E16. Associated with this change in polarity, vertically oriented fibers appear among the cells. These fibers can be traced to the inte


De novo transcriptome profiling of highly purified human lymphocytes primary cells.
To help better understand the role of long noncoding RNAs in the human immune system, we recently generated a comprehensive RNA-seq data set using 63 RNA samples from 13 subsets of T (CD4(+) naive, CD4(+) TH1, CD4(+) TH2, CD4(+) TH17, CD4(+) Treg, CD4(+) TCM, CD4(+) TEM, CD8(+) TCM, CD8(+) TEM, CD8(+) naive) and B (B naive, B memory, B CD5(+)) lymphocytes. There were five biological replicates for each subset except for CD8(+) TCM and B CD5(+) populations that included 4 replicates. RNA-Seq data were generated by an Illumina HiScanSQ sequencer using the TruSeq v3 Cluster kit. 2.192 billion of paired-ends reads, 2×100 bp, were sequenced and after filtering a total of about 1.7 billion reads were mapped. Using different de novo transcriptome reconstruction techniques over 500 previously unknown lincRNAs were identified. The current data set could be exploited to drive the functional characterization of lincRNAs, identify novel genes and regulatory networks associated with specific cells subsets of the human immune system.


A state-level application of the chronic illness breakthrough series: results from two collaboratives on diabetes in Washington State.
Breakthrough Series Collaboratives addressing chronic conditions have been conducted at the national level and in single health care delivery systems but not at the state level. Two state-level collaboratives were conducted: Diabetes Collaborative I (October 1999-November 2000) included 17 clinic teams from across the state, and Diabetes Collaborative II (February 2001-March 2002) included 30 teams and 6 health plans. Both collaboratives took place in Washington State, where a diverse group of primary care practices participated, and health insurance plans partnered with the clinic teams. Teams individually tested and implement changes in their systems of care to address all components of the Chronic Care Model. All 47 teams completed the collaboratives, and all but one maintained a registry throughout the 13 months. Most teams demonstrated some amount of improvement on process and outcome measures that addressed blood sugar testing and control, blood pressure control, lipid testing and control, foot exams, dilated eye exams, and self-management goals. The benefits of holding collaboratives more locally include increased technical support and increased participation, translating into wider implementation of prevention-focused, patient-centered care.


The effects of baclofen on the stretch reflex parameters of the cat.
Experiments were done in cats decerebrated at the precollicular postmammillary level to determine how a tonic increase of presynaptic inhibition of the intraspinal terminals of muscle spindle afferents changes the mechanical properties of the soleus stretch reflex (s.r.). Baclofen, a specific GABAB receptor agonist, was injected i.v. (1-2 mg/kg) so as to induce a tonic increase in presynaptic inhibition. The effects of baclofen on the stiffness and threshold of the s.r. were determined, respectively, from plots of stiffness vs background force and force vs length (length-tension plot). Baclofen, at these doses, had no effect on the excitation-contraction coupling properties of muscle or on the intrinsic stiffness-force relation. Changes of the soleus background force, required to obtain the stiffness vs force plots, were produced by stimulation of the contralateral common peroneal nerve or the posterior tibial nerve and occasionally by electrical stimulation in the area of the red nucleus. The stiffness of the s.r. as a function of the background force level was determined by stretching the muscle with a square pulse of 1-2 mm amplitude and 200-300 ms duration. The stiffness at each force level was calculated by dividing the change in force by the change in length, at a point where the force trace had stabilized. The length-tension relation of the s.r. was determined by stretching the muscle 12-17 mm at a constant rate of 1-2 mm/s. At all force levels, baclofen produced a significant decrease (40% or more) in the s.r. stiffness, within 10-15 min of injection as determined from the stiffness-force plots. The length-tension plus revealed that the decrease of s.r. stiffness was always accompanied by an increase in the s.r. threshold (typically 2-3 mm). It is suggested, therefore, that the s.r. threshold is not an independent variable, depending on the membrane potential of the alpha-motoneurons, and additionally on the level of presynaptic inhibition of the muscle spindle afferent terminals. The present results also imply that it may be possible for the CNS to adaptively modify the s.r. stiffness via presynaptic inhibition of the intraspinal terminals of muscle afferents. However, any such change of s.r. stiffness will be accompanied by a change in the s.r. threshold.


Clinical outcomes and survivorship of cementless triathlon total knee arthroplasties: a systematic review.
Over the last decade, cementless total knee arthroplasty has demonstrated improved outcomes and survivorship due to advances in technologies of implant design, manufacturing capabilities, and biomaterials. Due to increasing interest in cementless implant design for TKA, our aim was to perform a systematic review of the literature to evaluate the clinical outcomes and revision rates of the Triathlon Total Knee system over the past decade. A systematic review of the literature was conducted following PRISMA guidelines for patients who underwent total knee arthroplasty with cementless Triathalon Total Knee System implants. Patients had a minimum of two-year follow-up and data included clinical outcome scores and survivorship data. Twenty studies were included in the final analysis. The survivability of the Stryker Triathlon TKA due to all causes was 98.7%, with an aseptic survivability of 99.2%. The overall revision incidence per 1,000 person-years was 3.4. Re-revision incidence per 1,000 person-years was 2.2 for infection, and 1.3 for aseptic loosening. The average KSS for pain was 92.2 and the average KSS for function was 82.7. This systematic review demonstrated excellent clinical outcomes and survivorship at a mean time of 3.8 years. Additional research is necessary to examine the long-term success of the Stryker Triathlon TKA and the use of cementless TKAs in obese and younger populations. III.


Diagnostic Aspects of Vitamin D: Clinical Utility of Vitamin D Metabolite Profiling.
The assay of vitamin D that began in the 1970s with the quantification of one or two metabolites, 25-OH-D or 1,25-(OH)2D, continues to evolve with the emergence of liquid chromatography tandem mass spectrometry (LC-MS/MS) as the technique of choice. This highly accurate, specific, and sensitive technique has been adopted by many fields of endocrinology for the measurement of multiple other components of the metabolome, and its advantage is that it not only makes it feasible to assay 25-OH-D or 1,25-(OH)2D but also other circulating vitamin D metabolites in the vitamin D metabolome. In the process, this broadens the spectrum of vitamin D metabolites, which the clinician can use to evaluate the many complex genetic and acquired diseases of calcium and phosphate homeostasis involving vitamin D. Several examples are provided in this review that additional metabolites (eg, 24,25-(OH)2D3, 25-OH-D3-26,23-lactone, and 1,24,25-(OH)3D3) or their ratios with the main forms offer valuable additional diagnostic information. This approach illustrates that biomarkers of disease can also include metabolites devoid of biological activity. Herein, a case is presented that the decision to switch to a LC-MS/MS technology permits the measurement of a larger number of vitamin D metabolites simultaneously and does not need to lead to a dramatic increase in cost or complexity because the technique uses a highly versatile tandem mass spectrometer with plenty of reserve analytical capacity. Physicians are encouraged to consider adding this rapidly evolving technique aimed at evaluating the wider vitamin D metabolome toward streamlining their approach to calcium- and phosphate-related disease states. © 2021 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.


Structural, electronic, and magnetic properties of heterofullerene C(58)Si with odd number of atoms and a near planar tetracoordinate Si atom.
Density functional calculations and minimization techniques have been employed to characterize the structural and electronic properties of [5,6]-heterofullerene-C(58)Si-C(2v). Since it has odd number of atoms and a near planar tetracoordinate Si atom on the skeleton of the cage, it has odd number of atoms assembling a cage and is a novel molecule. Vibrational frequencies of the molecule have been calculated at the B3LYP/6-31G* level of theory. The absence of imaginary vibrational frequency confirms that the molecule corresponds to a true minimum on the potential energy hypersurface. Sixteen (13)C nuclear magnetic resonance (NMR) spectral signals of C(58)Si are characterized, and its heat of formation was estimated in this work.


Copper deficiency results in AMP-activated protein kinase activation and acetylCoA carboxylase phosphorylation in rat cerebellum.
Copper (Cu) deficiency impairs cerebellar development including biosynthetic processes like myelination and synaptogenesis. The activity of cerebellar mitochondrial cuproenzyme cytochrome c oxidase is markedly lower in Cu deficient rat pups and is accompanied by higher lactate levels indicating mitochondrial inhibition. Cu deficiency impaired energy metabolism is thought to contribute to developmental delays, but specific mechanisms linking these phenomena have remained unexplored. AMP-activated protein kinase (AMPK) is a cellular energy sensor that is activated during mitochondrial inhibition and shuts down biosynthetic processes to help conserve cellular ATP levels. Activated AMPK phosphorylates and inhibits acetylCoA carboxylase (ACC), the first enzyme in fatty acid biosynthesis. We hypothesize that AMPK is activated and ACC inhibited in Cu deficient cerebella. Perinatal copper deficiency was studied in young rats in rapidly frozen cerebella. Compared to copper-adequate (Cu+) pups, copper-deficient (Cu-) pups were hypothermic, had lower brain copper levels and markedly higher cerebellar lactate. Concentration of phosphorylated AMPK (pAMPK), indicating AMPK activation, was robustly higher in Cu- cerebella of rat pups at two ages and in two separate experiments. Compared to Cu+ cerebella, pACC content was significantly higher in all Cu- samples. Mechanisms leading to AMPK activation remain elusive. Higher AMP/ATP ratios and increased reactive nitrogen species (RNS) can lead to AMPK activation. ATP and AMP concentrations were unaltered and nitric oxide metabolites and 3-nitrotyrosine peptide levels remained unchanged in Cu- cerebella. AMPK activation may explain how ATP levels can be maintained even with a severe mitochondrial loss of CCO function.