Walking slope and heavy backpack loads affect torso muscle activity and kinematics.
The independent effects of sloped walking or carrying a heavy backpack on posture and torso muscle activations have been reported. While the combined effects of sloped walking and backpack loads are known to be physically demanding, how back and abdominal muscles adapt to walking on slopes with heavy load is unclear. This study quantified three-dimensional pelvis and torso kinematics and muscle activity from longissimus, iliocostalis, rectus abdominis, and external oblique during walking on 0° and ± 10° degree slopes with and without backpack loads using two different backpack configurations (hip-belt assisted and shoulder-borne). Iliocostalis activity was greater during downhill and uphill compared to level walking, but longissimus was only greater during uphill. Rectus abdominis activity was greater during downhill and uphill compared to level, while external oblique activity decreased as slopes progressed from down to up. Longissimus, but not iliocostalis, activity was reduced during both backpack configurations compared to walking with no pack. Hip-belt assisted load carriage required less rectus abdominis activity compared to using shoulder-borne only backpacks; however, external oblique was not influenced by backpack condition. Our results revealed different responses between iliocostalis and longissimus, and between rectus abdominis and external obliques, suggesting different motor control strategies between anatomical planes.


Contemporary Trends and Outcomes Associated With the Preprocedural Use of Oral P2Y12 Inhibitors in Patients Undergoing Percutaneous Coronary Intervention: Insights From the Blue Cross Blue Shield of Michigan Cardiovascular Consortium (BMC2).
We sought to describe trends in the use of preprocedural P2Y12 inhibitors and their clinical impact in patients undergoing percutaneous coronary intervention (PCI). Oral P2Y12 inhibitors are ubiquitously used medications; however, the specific timing of initial P2Y12 inhibitor administration remains intensely debated. Our study population comprised 74,053 consecutive patients undergoing PCI at 47 hospitals in Michigan from January 2013 through June 2015. In-hospital outcomes included stent thrombosis, bleeding, need for transfusion, and death. Hierarchical logistic regression, propensity matching, and targeted maximum likelihood estimation were used to adjust for baseline patient differences and clustering, and to minimize bias. Of 24,733 patients who received a preprocedural P2Y12 inhibitor, 82% received clopidogrel, 8% prasugrel, and 10% ticagrelor. Preprocedural administration of P2Y12 inhibitors declined during the study (49.3% to 24.8%; P<.001), and varied greatly across hospitals (14.5%-95.9%). No significant differences in outcomes were observed between patients receiving preprocedural clopidogrel and a matched cohort of those not receiving any preprocedural P2Y12 inhibitor (stent thrombosis: adjusted odds ratio [OR], 1.55; 95% confidence interval [CI], 0.30-7.84; bleeding: OR, 0.96; 95% CI, 0.63-1.46; transfusion: OR, 1.03; 95% CI, 0.69-1.55; and death: OR, 0.95; 95% CI, 0.38-2.37). Similar findings were demonstrated for preprocedural ticagrelor and prasugrel. Results from a subgroup analysis of patients with non-ST segment elevation acute coronary syndrome (n = 28,072) were consistent with the overall findings. There was a substantial decline in the rate of preprocedural P2Y12 inhibitor administration during the study. Furthermore, there were no significant differences in outcomes between patients treated with preprocedural P2Y12 inhibitors and those who were not.


Low-Temperature H2 Reduction of Copper Oxide Subnanoparticles.
Subnanoparticles (SNPs) with sizes of approximately 1 nm are attractive for enhancing the catalytic performance of transition metals and their oxides. Such SNPs are of particular interest as redox-active catalysts in selective oxidation reactions. However, the electronic states and oxophilicity of copper oxide SNPs are still a subject of debate in terms of their redox properties during oxidation reactions for hydrocarbons. In this work, in situ X-ray absorption fine structure (XAFS) measurements of Cu28 Ox SNPs, which were prepared by using a dendritic phenylazomethine template, during temperature-programmed reduction (TPR) with H2 achieved lowering of the temperature (T50 =138 °C) reported thus far for the CuII →CuI reduction reaction because of Cu-O bond elongation in the ultrasmall copper oxide particles.


Safety of repeat aprotinin administration for LVAD recipients undergoing cardiac transplantation.
Anecdotal reports of allergic and anaphylactic reactions after aprotinin therapy have raised concern that its repeat use may be associated with substantial morbidity. To address this concern, we reviewed our experience with all patients who underwent implantation of a left ventricular assist device and subsequent cardiac transplantation with perioperative use of aprotinin. Twenty-three patients received full-dose aprotinin during left ventricular assist device implantation and subsequent cardiac transplantation. All patients tolerated primary exposure to aprotinin without complication. One episode of anaphylaxis after secondary exposure was treated with rapid institution of cardiopulmonary bypass. Although renal dysfunction was observed shortly after cardiac transplantation in 30.4% of patients, the effect was transient and occurred in the presence of cyclosporine. The one perioperative death after secondary exposure was unrelated to bleeding complications. No clinically evident thromboembolic events were documented. Primary and secondary exposure to aprotinin during operation with cardiopulmonary bypass is associated with limited intraoperative blood use, a low incidence of transient renal dysfunction and anaphylaxis, a rare need of reoperation for bleeding, and no clinical thromboembolic events.


Using Analogies to Help Adolescents Unlearn Self-Blame.
One of the changes included in DSM-5 to the diagnosis of posttraumatic stress disorder (PTSD) was the inclusion of negative cognitions and emotions related to the trauma, including distorted self-blame for the trauma.1 Studies show that many young people experience self-blame after experiencing various types of trauma,2-4 that self-blame from trauma is a key mediating factor in internalizing symptoms in adolescents,2 and that self-blame is tied to higher levels of trauma symptoms.3 However, studies have also suggested that young people develop self-blame when they do not directly experience PTSD from trauma, such as maternal depression, parental divorce or separation, bullying, and general negative events.5 There appear to be multiple pathways in which young people learn self-blame, and self-blame should be assessed on an individualized level. It has been suggested that psychotherapy with children and adolescents should involve translating the goals of treatment into simple concrete concepts, such as "a simple situation that is familiar to them from day-to-day life."6 In addition to evidence-based psychotherapeutic interventions that target symptoms of self-blame resulting from trauma, analogies may serve as potential intervention tools to translate the goal of unlearning self-blame relevant to the individual, situational, and cultural contexts.


Genetic Variations of ferroportin-1(FPN1-8CG), TMPRSS6 (rs855791) and Hemojuvelin (I222N and G320V) Among a Cohort of Egyptian β-Thalassemia Major Patients.
Iron overload remains a major cause of morbidity and mortality among β-thalassemia major (β-TM) patients. Iron regulatory proteins and their genetic variants together with changes in hepcidin levels in thalassemic patients could affect the disease manifestations. This work aimed to study genetic variations of ferroportin-1 (FPN1-8CG), Transmembrane Serine Protease 6 (TMPRSS6 rs855791) and hemojuvelin (HJV I222N and G320V) genes within a cohort of 97 β-TM Egyptian patients by Polymerase chain reaction Restriction Fragment Length Polymorphism (PCR-RFLP) in comparison to fifty normal control subjects. Among β-TM patients; the CG variant of FPN1 was significantly higher, while the TT and TC variants of TMPRSS6 were significantly lower in comparison to controls. Liver Iron Concentration (LIC) was significantly higher among β-TM patients harboring the FPN1 (GG) genotype and we found that FPN1gene mutation acts as independent predictor of MRI LIC (p = 0.011), Pulmonary artery pressure (PAP) was significantly higher in patients harboring the mutant FPN1 (GG and CG) genotypes (p value 0.04). β-TM patients having the HJV I222N (AA) genotype were having significantly higher cardiac iron overload (p value = 0.026). The studied genetic variants of iron regulatory proteins could alter the manifestations of iron overload thus resulting in different clinical phenotypes of thalassemic patients, these findings need to be confirmed by larger cohorts of patients with longer follow-up periods. The online version contains supplementary material available at 10.1007/s12288-022-01580-8.


Effects of quercetin and quercetin-3-O-glycosides on oxidative damage in rat C6 glioma cells.
Flavonoids are reported to be powerful antioxidants in cell free systems. They naturally occur as glycosides rather than as aglycon. In this study the ability of the flavonoid quercetin and its glycosides, quercetin-3-O-rutinoside (rutin), quercetin-3-O-glucoside and quercetin-3-O-(6″-O-acetyl)-glucoside, to protect in vitro rat C6 glioma cells from oxidative damage induced by cumene hydroperoxide was investigated. Cumene hydroperoxide induced cell death and lipid peroxidation. The cytotoxicity of cumene hydroperoxide could be prevented by the radical scavenger dimethyl thiourea and the ferric iron chelator deferoxamine, indicating that its cytotoxic activity is related to the generation of reactive oxygen radicals in the ferrous iron dependent Fenton reaction. Quercetin, in a concentration range of 10-100 μM, but neither rutin nor the other two glycosides, were able to protect C6 cells from cytotoxicity and lipid peroxidation. Furthermore, cytoprotective concentrations of quercetin proved to be cytotoxic itself. These results call in question potential beneficial effects of dietary intake or therapeutic use of naturally occurring flavonoids.


Multicentre, clinical trial of burst spinal cord stimulation for neck and upper limb pain NU-BURST: a trial protocol.
Spinal cord stimulation (SCS) is an established therapy for chronic neuropathic pain and most frequently utilised for Failed Back Surgery Syndrome (FBSS). BurstDR™ also known as DeRidder Burst-SCS, a novel waveform, has demonstrated superiority to conventional tonic stimulation of the thoracic spine in FBSS. There are case reports of an improvement in multidimensional pain outcomes using DeRidder Burst-SCS in the cervical spine for chronic neck and cervical radicular pain. The safety and efficacy of cervical DeRidder Burst-SCS stimulation still however remain undetermined. This is a prospective, multicentre feasibility trial evaluating the safety and therapeutic efficacy of DeRidder Burst-SCS stimulation for the treatment of chronic intractable neck pain with or without radiation to the arm, shoulder, and upper back. After baseline evaluation, subjects will undergo an SCS trial using the Abbott Invisible Trial system according to standard clinical procedures. During the trial phase, SCS leads will be implanted in the cervical epidural space. At the end of the SCS trial, subjects experiencing at least 50% pain relief will be considered for permanent implant. Pain intensity, medication usage, and other multidimensional pain outcomes will be collected. The timing of these will be at baseline, end of the SCS trial and at 3-, 6-, and 12-month visits. Incidence of adverse events will be collected throughout the study duration. The results of this feasibility study will validate the efficacy and safety of DeRidder Burst-SCS stimulation in the cervical spine. The results obtained in this study will potentially be used to generate a level 1 evidence-based study with formal statistical hypotheses testing. www.clinicaltrials.gov Identifier: NCT03159169.


Comparative anatomical studies of the SmL face cortex with special reference to the occurrence of "barrels" in layer IV.
In the SmL cortex of mice and rats there are cytoarchitestonically identificable groups of cells -- called barrels -- some of which have been shown to be directly related to whiskers and other sensory hairs on the contralateral face. In this study we have used a comparative approach to determine the incidence and variation of the barrels. The brains of 27 mammalian species have been examined histologically to determine whether barrels exist in layer IV of what is known or likely to be the face area of SmI. Thick sections (50-100 mum) were taken tangential to the pia overlying SmI and stained with thionin. The patterns of facial whiskers were also mapped by dissection of the facial skin. Barrels were seen only in brains of species belonging to three of the seven mammalian orders examined. We have confirmed Weller's ('72) observation of barrels in the Australian brush-tailed possum but have not found barrels in two marsupials from the western hemisphere. Barrels were demonstrable in representatives of four of five rodent suborders examined and in the rabbit. From the study of the rodent brains, a number of trends emerge. (1) The organization of the barrel fields is "dictated" by the organization of the sensory periphery. Animals with five rows of large mystacial (moustache-like) vibrissae have five rows of PMBSF (Posteromedial barrel sub-field) barrels. (2) The barrels are confined to layer IV of (what is known or likely to be) the SmI face area. The pattern and cortical location of the barrel field is consistent among different specimens of the same species. (3) Certain behavioral patterns do not preclude the existence of the barrels. Species which possess well developed visual systems and behaviors (e.g., grey squirrel) and forms which do not actively explore the environment by whisking their vibrissae (e.g., guinea pig) have barrels. (4) Within a given rodent suborder, the barrels become more difficult to identify, as the brains become larger. We have not yet been able to demonstrate barrels in the largest rodent, the capybara.


Non-Necrotizing Soft Tissue Infection and Streptococcal Toxic Shock Syndrome Caused by a Novel Streptococcus pyogenes Subtype (emm76.10).
We report a case of non-necrotizing soft tissue infection and streptococcal toxic shock syndrome (STSS) caused by a novel emm subtype (emm76.10) of group A Streptococcus pyogenes (GAS). A 54-year-old Japanese woman suffering from fever, fatigue, and lower abdominal pain along with erythema for 3 days was admitted to our hospital. Additionally, she presented with hypotension and multiple organ failure. Exploratory incision was performed due to the presence of STSS and for an examination of the necrotizing soft tissue infection from her lower abdomen to the left thigh. Tissue cultures from the exudates and fascia yielded positive results for GAS growth, although blood cultures returned as negative. After 15 days of antimicrobial therapy, she recovered fully without any complications. Genotyping of the isolate indicated a novel emm subtype (emm76.10), with 5 amino acid substitutions in the emm76.0 subtype sequence and the full-length sequence of 780 bp. This isolate was resistant to tetracyclines, macrolides-lincosamides, and fluoroquinolones, owing to the presence of antibiotic resistance determinants, tet(M) and erm(B), and point mutations, Ser79Phe/Ser81Phe, in the quinolone resistance-determining regions of parC/gyrA. In conclusion, our observations suggest the importance of early-stage exploratory incision and drainage from the infected region for the isolation and characterization of causative bacteria to facilitate selection of appropriate antibiotics for treatment.


Subdivision of the tribe Oligaphorurini in the light of new and lesser known species from North-East Russia (Collembola, Onychiuridae, Onychiurinae).
The paper is devoted to a taxonomic review of Oligaphorurini from the north-eastern part of Palaearctic. Two new species, i.e. Oligaphoruraambigua sp. n. and Oligaphoruraduocellata sp. n., are described. Four species, Oligaphoruranataliae (Fjellberg, 1987), Oligaphorurainterrupta (Fjellberg, 1987), Oligaphorurapingicola (Fjellberg, 1987), and Micraphoruraalnus (Fjellberg, 1987), are redescribed on base of the types and new material, and remarks on other species known for the region, Oligaphoruragroenlandica (Tullberg, 1876), Oligaphoruraursi (Fjellberg, 1984), Oligaphoruraaborigensis (Fjellberg, 1987), and Micraphoruraabsoloni (Börner, 1901), are given to clarify their generic affiliation. Finally, merits and disadvantages of the current subdivision of the tribe are discussed and a key to the northern species of the tribe is provided.


"Desert" gene (Chr9p21) variants as novel markers for coronary artery disease.
Previous reports have denoted to the possible link of Chr9p21 locus to the incidence of coronary artery disease (CAD). The entire core of chr9p21 is covered by "ANRIL" (Antisense noncoding RNA in INK4 Locus) and lies in a region that is free from any coding proteins; therefore, it is called the desert gene. The major objectives of this study were to examine the association of rs10757278 and rs2383206 SNPs on Chr9p21 with the incidence of CAD in the presence and absence of type 2 diabetes (T2D) in Egyptians and to correlate these genetic variants with several disease biomarkers (TC, CRP, and HbA1c). The study subjects consisted of 150 subjects; 50 healthy controls and 100 patients that were divided into two groups; CAD patients and CAD T2D patients. The genotyping of SNPs was performed using qPCR. Genotype distribution for both SNPs were found to be significantly different (p=0.0009 for rs10757278 and p=0.001 for rs2383206) between patients and controls. The allele frequency was also different for rs10757278. The current study showed that rs10757278/rs2383206-G allele increases the risk for CAD in Egyptians. Moreover, AA variant appeared as a protective genotype. However, SNPs did not noticeably contribute in the elevation of TC, hs-CRP, and HbA1c in non-diabetic and diabetic CAD patients.


Evaluation of five cell types for the isolation of herpes simplex virus.
Five cells were evaluated in a comparative analysis for sensitivity, specificity, and rapidity in detecting the presence of herpes simplex virus HSV-1 and HSV-2. Included in this study were human embryonic kidney (HEK), rabbit kidney (RK), MRC-5, mink lung (ML), and Microtus agrestis (UMMA). A total of 274 specimens from genital, throat, skin, or other sources that were submitted for HSV isolation were used in the study. The sensitivity of the different cells was assessed by the total number of positive cultures detected by all the cells under evaluation. At 48 hr, HEK and RK detected 80% of the positives, ML detected 79%, MRC-5 detected 73%, and UMMA detected 60%. All cells tested were satisfactory; however, the choice of which cell to use for isolation of HSV depends upon the needs of the specific laboratory.


Direct-injection chemiluminescence detector. Properties and potential applications in flow analysis.
We present a novel chemiluminescence detector, with a cone-shaped detection chamber where the analytical reaction takes place. The sample and appropriate reagents are injected directly into the chamber in countercurrent using solenoid-operated pulse micro-pumps. The proposed detector allows for fast measurement of the chemiluminescence signal in stop-flow conditions from the moment of reagents mixing. To evaluate potential applications of the detector the Fenton-like reaction with a luminol-H2O2 system and several transition metal ions (Co(2+), Cu(2+), Cr(3+), Fe(3+)) as a catalyst were investigated. The results demonstrate suitability of the proposed detector for quantitative analysis and for investigations of reaction kinetics, particularly rapid reactions. A multi-pumping flow system was designed and optimized. The developed methodology demonstrated that the shape of the analytical signals strongly depends on the type and concentration of the metal ions. The application of the detector in quantitative analysis was assessed for determination of Fe(III). The direct-injection chemiluminescence detector allows for a sensitive and repeatable (R.S.D. 2%) determination. The intensity of chemiluminescence increased linearly in the range from about 0.5 to 10 mg L(-1) Fe(III) with the detection limit of 0.025 mg L(-1). The time of analysis depended mainly on reaction kinetics. It is possible to achieve the high sampling rate of 144 samples per hour.


Minimally Invasive Robotic-Assisted Patellofemoral Arthroplasty.
Isolated patellofemoral arthritis is a common debilitating condition in adults older than 40 years of age. Surgical options such as patellofemoral arthroplasty exist for those who failed to respond to nonoperative treatment. However, early patellofemoral arthroplasty techniques often resulted in poor outcomes due to mal-tracking and malalignment of components. Robotic-assisted surgery recently has been introduced as an alternative to classic patellofemoral arthroplasty, with the potential to improve the anatomical fit and reproducibility of implant positioning. We present the technique for minimally invasive robotic-assisted patellofemoral arthroplasty system.


25 years since the eradication of smallpox: why poxvirus research is still relevant.
The World Health Organization (WHO) announced the eradication of smallpox twenty-five years ago this month. This conquest of an infectious disease, which has been the bane of humankind for centuries, still stands as the WHO's greatest achievement. The anniversary of such a scientific and medical landmark provides an appropriate occasion to reflect on this feat and to assess the significance and necessity of the poxvirus research that has followed this.


A novel thermostable hemoglobin from the actinobacterium Thermobifida fusca.
The gene coding for a hemoglobin-like protein (Tf-trHb) has been identified in the thermophilic actinobacterium Thermobifida fusca and cloned in Escherichia coli for overexpression. The crystal structure of the ferric, acetate-bound derivative, was obtained at 2.48 A resolution. The three-dimensional structure of Tf-trHb is similar to structures reported for the truncated hemoglobins from Mycobacterium tuberculosis and Bacillus subtilis in its central domain. The complete lack of diffraction patterns relative to the N- and C-terminal segments indicates that these are unstructured polypeptides chains, consistent with their facile cleavage in solution. The absence of internal cavities and the presence of two water molecules between the bound acetate ion and the protein surface suggest that the mode of ligand entry is similar to that of typical hemoglobins. The protein is characterized by higher thermostability than the similar mesophilic truncated hemoglobin from B. subtilis, as demonstrated by far-UV CD melting experiments on the cyano-met derivatives. The ligand-binding properties of Tf-trHb, analyzed in stopped flow experiments, demonstrate that Tf-trHb is capable of efficient O2 binding and release between 55 and 60 degrees C, the optimal growth temperature for Thermobifida fusca.


New antibodies immobilization system into a graphite-polysulfone membrane for amperometric immunosensors.
Polysulfone membrane is used for the first time for the preparation of electrochemical immunosensors. A disposable immunosensor based on a porous conductor polymer graphite-polysulfone-electrode has been developed using a phase inversion technique for the determination of anti-rabbit IgG (anti-RIgG) as a model analyte. To construct the sensor, a conductor membrane was deposited on the surface of working graphite-epoxy composite (GEC) electrode. The membrane was characterized by SEM. This sensor was based on the competitive assay between free and labeled anti-RIgG for the available binding sites of immobilized rabbit IgG (RIgG). Incubation parameters were optimized in this work. The immunological reaction was detected using an enzymatic-labeling procedure (HRP enzyme) combined with the amperometric detection using H(2)O(2) as substrate and hydroquinone as mediator. This sensor shows stability during a week and a good reproducibility. The current was monitored amperometrically at -0.1 V versus SCE and this method showed a linear range of the anti-RIgG from 1 to 6 microg/ml. The detection limit was determined to be 0.77 microg/ml.


Traversing the diverse avenues of exopolysaccharides-based nanocarriers in the management of cancer.
Exopolysaccharides are unique polymers generated by living organisms such as algae, fungi and bacteria to protect them from environmental factors. After a fermentative process, these polymers are extracted from the medium culture. Exopolysaccharides have been explored for their anti-viral, anti-bacterial, anti-tumor, and immunomodulatory effects. Specifically, they have acquired massive attention in novel drug delivery strategies owing to their indispensable properties like biocompatibility, biodegradability, and lack of irritation. Exopolysaccharides such as dextran, alginate, hyaluronic acid, pullulan, xanthan gum, gellan gum, levan, curdlan, cellulose, chitosan, mauran, and schizophyllan exhibited excellent drug carrier properties. Specific exopolysaccharides, such as levan, chitosan, and curdlan, have demonstrated significant antitumor activity. Moreover, chitosan, hyaluronic acid and pullulan can be employed as targeting ligands decorated on nanoplatforms for effective active tumor targeting. This review shields light on the classification, unique characteristics, antitumor activities and nanocarrier properties of exopolysaccharides. In addition, in vitro human cell line experiments and preclinical studies associated with exopolysaccharide-based nanocarriers have also been highlighted.


The opportunistic pathogen Stenotrophomonas maltophilia utilizes a type IV secretion system for interbacterial killing.
Bacterial type IV secretion systems (T4SS) are a highly diversified but evolutionarily related family of macromolecule transporters that can secrete proteins and DNA into the extracellular medium or into target cells. It was recently shown that a subtype of T4SS harboured by the plant pathogen Xanthomonas citri transfers toxins into target cells. Here, we show that a similar T4SS from the multi-drug-resistant opportunistic pathogen Stenotrophomonas maltophilia is proficient in killing competitor bacterial species. T4SS-dependent duelling between S. maltophilia and X. citri was observed by time-lapse fluorescence microscopy. A bioinformatic search of the S. maltophilia K279a genome for proteins containing a C-terminal domain conserved in X. citri T4SS effectors (XVIPCD) identified twelve putative effectors and their cognate immunity proteins. We selected a putative S. maltophilia effector with unknown function (Smlt3024) for further characterization and confirmed that it is indeed secreted in a T4SS-dependent manner. Expression of Smlt3024 in the periplasm of E. coli or its contact-dependent delivery via T4SS into E. coli by X. citri resulted in reduced growth rates, which could be counteracted by expression of its cognate inhibitor Smlt3025 in the target cell. Furthermore, expression of the VirD4 coupling protein of X. citri can restore the function of S. maltophilia ΔvirD4, demonstrating that effectors from one species can be recognized for transfer by T4SSs from another species. Interestingly, Smlt3024 is homologous to the N-terminal domain of large Ca2+-binding RTX proteins and the crystal structure of Smlt3025 revealed a topology similar to the iron-regulated protein FrpD from Neisseria meningitidis which has been shown to interact with the RTX protein FrpC. This work expands our current knowledge about the function of bacteria-killing T4SSs and increases the panel of effectors known to be involved in T4SS-mediated interbacterial competition, which possibly contribute to the establishment of S. maltophilia in clinical and environmental settings.


Cortical tubers, cognition, and epilepsy in tuberous sclerosis.
Tuberous sclerosis complex is an autosomal-dominant genetic disorder characterized by hamartomatous growth in various organs. Patients who have this disorder exhibit a high rate of epilepsy and cognitive problems. We investigated number of tubers, location, seizure types, and cognitive outcome, and we analyzed the relationships among them in our tuberous sclerosis patients in the Comprehensive Epilepsy Program at the University of Alberta. We also examined the seizure outcome after tuber resection. Our study cohort included 24 patients with tuberous sclerosis complex. We obtained seizure history, electroencephalogram, and neuropsychologic parameters. Magnetic resonance imaging was used to examine tuber numbers and locations. Ten patients underwent surgical removal of tubers responsible for intractable epilepsy. A negative correlation was found between the number of tubers and intelligent quotient score. Epilepsy surgery led to freedom from seizures in this patient group. We demonstrated that the total number and location of cortical tubers play a significant role in the extent of mental retardation in patients with tuberous sclerosis complex. In addition, patients with intractable seizures and well-defined epileptic focus had excellent surgical outcome.


Infection of mouse bone marrow-derived dendritic cells by live attenuated Japanese encephalitis virus induces cells maturation and triggers T cells activation.
An attenuated Japanese encephalitis virus (JEV) strain SA14-14-2, generated from the wild strain SA14, is an effective live vaccine against JEV infection. It has led to a significant decrease in JEV infection around the world. Although it is highly effective, the mechanism for its robust immunity was not well investigated. In this study, the interaction of SA14-14-2 with bone marrow-derived dendritic cells (bmDCs) was investigated. Our results showed that the infection of bmDCs with SA14-14-2 resulted in viral replication and upregulation of bmDC maturation marker molecules (CD40, CD80, CD83 and MHC I). SA14-14-2 infection also stimulated the production of interferon-α (IFN-α), monocyte chemoattractant protein-1 (MCP-1/CCL2), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) of bmDC. Both MLR and ELISPOT assay showed an enhanced allostimulatory capacity of SA14-14-2-infected bmDCs. Furthermore, the SA14-14-2-infected bmDCs impaired the expansion of Foxp3+ regulatory T (Treg) cells with immunosuppressive potential, suggesting that SA14-14-2 infection induced antiviral immunity rather than immunosuppression. Taken together, our results indicated that SA14-14-2 infection caused bmDC maturation, changed the expression profiles of several cytokines, and triggered T cell activation. This offered an insight in the immunologic mechanisms associated with the high efficiency of the SA14-14-2 vaccine.


Time trends in socioeconomic inequalities in cancer mortality: results from a 35 year prospective study in British men.
Socioeconomic inequalities in cancer mortality in Britain have been shown to be present in the 1990s and early 2000s. Little is known about on-going patterns in such inequalities in cancer mortality. We examined time trends in socioeconomic inequalities in cancer mortality in Britain between 1978 and 2013. A socially representative cohort of 7489 British men with data on longest-held occupational social class, followed up for 35 years, in whom 1484 cancer deaths occurred. The hazard ratio for cancer mortality for manual vs. non-manual social classes remained unchanged; among men aged 50-59 years it was 1.62 (95%CI 1.17-2.24) between 1980-1990 and 1.65 (95%CI 1.14-2.40) between 1990-2000. The absolute difference (non-manual minus manual) in probability of surviving death from cancer to 70 years remained at 3% over the follow-up. The consistency of risks over time was similar for both smoking-related and non-smoking related cancer mortality. Socioeconomic inequalities in cancer mortality in Britain remain unchanged over the last 35 years and need to be urgently addressed.


Outcomes of a Pharmacist-Led Hepatitis C Virus Treatment Program in an Urban Safety-Net Health System, Chicago, 2017-2019.
A recommendation in March 2020 to expand hepatitis C virus (HCV) screening to all adults in the United States will likely increase the need for HCV treatment programs and guidance on how to provide this service for diverse populations. We evaluated a pharmacist-led HCV treatment program within a routine screening program in an urban safety-net health system in Chicago, Illinois. We collected data on all patient treatment applications submitted from January 1, 2017, through June 30, 2019, and assessed outcomes of and patient retention in the treatment cascade. During the study period, 203 HCV treatment applications were submitted for 187 patients (>1 application could be submitted per patient): 49% (n = 91) were aged 55-64, 62% (n = 116) were male, 67% (n = 125) were Black, and 15% (n = 28) were Hispanic. Of the 203 HCV treatment applications, 87% (n = 176) of patients were approved for treatment, 91% (n = 161) of whom completed treatment. Of the 161 patients who completed treatment, 81% (n = 131) attended their sustained virologic response (SVR) follow-up visit, 98% (n = 129) of whom reached SVR. The largest drop in the treatment cascade was the 19% decline from receipt of treatment to SVR follow-up visit. The pharmacist-led model for HCV treatment was effective in navigating patients through the treatment cascade and achieving SVR. Widespread implementation of pharmacist-led HCV treatment models may help to hasten progress toward 2030 HCV elimination goals.


Genetics of affective (mood) disorders.
The enormous public health importance of mood disorders, when considered alongside their substantial heritabilities, has stimulated much work, predominantly in bipolar disorder but increasingly in unipolar depression, aimed at identifying susceptibility genes using both positional and functional molecular genetic approaches. Several regions of interest have emerged in linkage studies and, recently, evidence implicating specific genes has been reported; the best supported include BDNF and DAOA but further replications are required and phenotypic relationships and biological mechanisms need investigation. The complexity of psychiatric phenotypes is demonstrated by (a) the evidence accumulating for an overlap in genetic susceptibility across the traditional classification systems that divide disorders into schizophrenia and mood disorders, and (b) evidence suggestive of gene-environment interactions.


Vanadyl (IV) and vanadate (V) binding to selected endogenous phosphate, carboxyl, and amino ligands; calculations of cellular vanadium species distribution.
Vanadium enters cells as vanadate (V) where it is reduced to vanadyl (IV), VO2+. Vanadate species at plasma pH, H2VO4-, and HVO4(2-) are referred to as VO3-. To gain an insight into the subcellular vanadium distribution we measured the binding of VO3- and VO2+ to extra- and intracellular ligands, and calculated free and bound fractions of these ions for expected in vivo conditions. The association constants (K) were determined by the pH shift caused by an addition of VOSO4 or NaVO3 to individual ligand solutions at 20 degrees C and a pH equal to the pK of the reactive groups. The pk's for binding of VO2+ were ATP, 5.9; ADP, 5.5; AMP, 5.1; Pi 4.3; creatine phosphate (CP), 3.6; glutamic acid, 3.4; aspartic acid, 3.1; human serum albumin, 3.1; glutathione, 2.7; ascorbic acid, 3.3; citric acid, 4.0. The pk of VO3- and human serum albumin was 3.3 and of that VO3- and glutathione was 4.2. VO3- did not bind to ATP, even via Mg2+ or Ca2+ bridges. We calculated that in cells approximately 1% of total VO2+ is unbound, which is 10(-10)-10(-9) M since published values for total vanadium (mainly VO2+) concentrations in tissues are on the order of 10(-8)-10(-7) M. Free VO2+ may be even less because of binding to additional ligands not considered and due to spontaneous hydrolysis to VOOH+ and VO(OH)2(2+) at intracellular pH. The binding of VO2+ to each ligand was corrected for presence of multiple ligands and competition by H+, K+, and Mg2+. In cells with no CP, up to 70% of VO2+ is bound to phosphates and up to 29% to proteins; in cells with 30 mM CP (as in muscle), approximately 95% is bound to phosphates (CP binds up to 61% of total VO2+) and approximately 4% to proteins; in cells with 2 mM ascorbic acid (as in brain), the vitamin binds approximately 3% of total VO2+. These binding values apply for the total VO2+ concentration range of 10(-8)-10(-5) M. The intracellular binding and a reducing environment protect the freshly reduced VO2+ from oxidation to VO3- that would otherwise occur at neutral pH. This strong affinity of VO2+ primarily for phosphates also explains the mechanism for the intracellular accumulation of vanadium which is a factor in previously observed transport of VO3- into cells.(ABSTRACT TRUNCATED AT 400 WORDS)


"Azole menace"-An underappreciated trigger perpetuating the epidemic of antifungal therapeutic failure in cutaneous mycoses.
The South-Asian epidemic of anti-fungal therapeutic failures (AFTF) is on the rise. Although many demographic, environmental, and socioeconomic factors have been implicated in the genesis of this problem, two pharmacological issues warrant attention. While detailed discussions on the role of topical corticosteroid (TCS) in the changing landscape of the superficial mycotic infections in this region have been making headlines, another equally, rather more important pharmacological factor seems to have been undermined by the hype around TCS. The fastidious pharmacokinetic properties and related practical aspects of the triazole group of oral and topical antifungals, especially oral itraconazole seem to contribute significantly to the persistence of AFTF epidemic. In this paper, we shall discuss the broad aspects of the spectral precariousness of oral triazole antifungals with special emphasis to itraconazole, a concept known as the "azole menace" in the overall pathogenesis and tenacity of the AFTF epidemic.


Flexibility between the protease and helicase domains of the dengue virus NS3 protein conferred by the linker region and its functional implications.
The dengue virus (DENV) NS3 protein is essential for viral polyprotein processing and RNA replication. It contains an N-terminal serine protease region (residues 1-168) joined to an RNA helicase (residues 180-618) by an 11-amino acid linker (169-179). The structure at 3.15 A of the soluble NS3 protein from DENV4 covalently attached to 18 residues of the NS2B cofactor region (NS2B(18)NS3) revealed an elongated molecule with the protease domain abutting subdomains I and II of the helicase (Luo, D., Xu, T., Hunke, C., Grüber, G., Vasudevan, S. G., and Lescar, J. (2008) J. Virol. 82, 173-183). Unexpectedly, using similar crystal growth conditions, we observed an alternative conformation where the protease domain has rotated by approximately 161 degrees with respect to the helicase domain. We report this new crystal structure bound to ADP-Mn(2+) refined to a resolution of 2.2 A. The biological significance for interdomain flexibility conferred by the linker region was probed by either inserting a Gly residue between Glu(173) and Pro(174) or replacing Pro(174) with a Gly residue. Both mutations resulted in significantly lower ATPase and helicase activities. We next increased flexibility in the linker by introducing a Pro(176) to Gly mutation in a DENV2 replicon system. A 70% reduction in luciferase reporter signal and a similar reduction in the level of viral RNA synthesis were observed. Our results indicate that the linker region has evolved to an optimum length to confer flexibility to the NS3 protein that is required both for polyprotein processing and RNA replication.


Clinical profile of acute pancreatitis following treatment with protease inhibitors: a real-world analysis of post-marketing surveillance data.
Acute pancreatitis (AP) has been reported in patients treated with protease inhibitors (PIs), but there are few real-world studies comparing the occurrence and characteristics of AP after different PI regimens. Disproportionality analysis and Bayesian analysis were utilized for data mining of the Food and Drug Administration's Adverse Event Reporting System (FAERS) database for suspected adverse events involving AP after PI. The times to onset and fatality rates of AP following different PI regimens were also compared. Based on 33,832 reports related to PIs, 285 cases were associated with AP, involving with 12 out of the 15 studied PIs. Of all the reported AP events related to PIs, 64.56% occurred in men and the median time to onset of AP was 103 (IQR: 26-408) days after the initiation of PI treatment with a fatality rate of 14.02%. Among all PI therapies, indinavir was notably associated with AP, and ritonavir and lopinavir/ritonavir-induced AP cases appeared to be associated with a higher risk of death. Most of PIs were associated with AP-related adverse events, among which indinavir has a stronger association with AP but there is no significant difference in fatality rates.


Autoantigen Ro52 is an E3 ubiquitin ligase.
Anti-Ro/SSA antibodies are classic autoantibodies commonly found in patients with Sjögren's syndrome, a chronic autoimmune disease characterized by dryness of the eyes and mouth. The autoantibodies recognize a RING-finger protein, Ro52, whose function is still unknown. Since many RING-finger proteins have been identified as E3 ubiquitin ligases, this study was designed to determine whether Ro52 functions as an E3 ubiquitin ligase. For this purpose, recombinant Ro52 was purified from bacterial lysate and used to investigate its activity of E3 ubiquitin ligase in vitro. Its enzymatic activity was also tested in HEK293T cells using wild-type Ro52 and its RING-finger mutant. Our results indicated that Ro52 ubiquitinates itself in cooperation with E2 ubiquitin-conjugating enzyme UbcH5B, thereby validating that Ro52 is a RING-finger-type E3 ubiquitin ligase. Importantly, this ubiquitin modification is predominantly monoubiquitination, which does not target Ro52 to the proteasome for degradation.


Multislice computed tomography for prediction of optimal angiographic deployment projections during transcatheter aortic valve implantation.
This study assessed whether multislice computed tomography (MSCT) could predict optimal angiographic projections for visualizing the plane of the native valve and facilitate accurate positioning during transcatheter aortic valve implantation (TAVI). Accurate device positioning during TAVI depends on valve deployment in angiographic projections perpendicular to the native valve plane, but these may be difficult to determine. Twenty patients underwent MSCT before TAVI. Using a novel technique, multiple angiographic projections accurately representing the native valve plane in multiple axes were determined. The accuracy of all predicted projections was determined post-procedure using angiography according to new criteria, based on valve perpendicularity and the degree of strut overlap (defined as excellent, satisfactory, or poor). The accuracy of valve deployment using MSCT was compared with the results of 20 consecutive patients undergoing TAVI without such MSCT angle prediction. Correct final deployment projections were more frequent in the MSCT-guided compared with non-MSCT-guided group: excellent or satisfactory projections (90% vs. 65%, p = 0.06). The MSCT angle prediction was accurate but dependent on optimal images (optimal images: 93% of predicted angles were excellent or satisfactory, suboptimal images: 73% of predicted angles were poor). A "line of perpendicularity" could be generated with optimal projections across the right-to-left anterior oblique plane by adding the correct cranial or caudal angulation. Pre-procedural MSCT can predict optimal angiographic deployment projections for implantation of transcatheter valves. An ideal deployment angle curve or "line of perpendicularity" can be generated. Understanding and applying these principles improves the accuracy of valve deployment and may improve outcomes.


Effectiveness of gabapentin in controlling spasticity: a quantitative study.
The purpose of this investigation was to study the effectiveness of gabapentin in controlling spasticity in persons with spinal cord injury (SCI) using a surface EMG-based quantitative assessment technique called the brain motor control assessment (BMCA). Six men from a Veterans Affairs Medical Center with spasticity due to traumatic SCI were studied as part of a multi-center, placebo-controlled, cross-over, clinical trial of gabapentin. Spasticity was evaluated using multi-channel surface EMG recordings of muscles in the lower extremities, abdomen and low back before and during treatment with oral gabapentin or placebo. Gabapentin or placebo was given orally in doses 400 mg three times daily for 48 h. Following a 10 day wash-out period subjects were crossed-over to receive the medication not received the first time. This was followed by an elective open-label extension. Group results during the controlled trial did not reach statistical significance at the dosage used. One subject demonstrated a dramatic improvement in spasticity that was apparent both clinically and with the BMCA. Other subjects demonstrated modest improvements which were seen in the BMCA but not recognized clinically. During the open label extension, the four subjects who participated experienced important clinical improvements with higher doses (to 3600 mg/day). These improvements were often in components of spasticity in which the BMCA had detected subclinical changes during the cross-over trial. A seventh subject was studied using the BMCA at doses of 1200 mg T.I.D. gabapentin, off gabapentin and 800 mg T.I.D. gabapentin and demonstrated quantitatively a dose-related effect with higher doses of gabapentin which matched clinical observations. Gabapentin at doses of 400 mg T.I.D. may be effective in controlling some features of spasticity in persons with SCI. Higher doses provide greater control of spasticity, and controlled studies using higher doses are needed to evaluate gabapentin's efficacy.


Crystallization of the maltodextrin-binding protein for active transport and chemotaxis in several different liganded and mutant forms.
We have crystallized the maltodextrin-binding protein complexed with two cyclic maltodextrins, and with no ligand at all. We have also grown crystals of two complexes of a mutant of maltodextrin-binding protein whose transport specificity has been altered. Studies of these crystals should provide considerable information about protein-oligosaccharide interactions, the effects of these interactions on oligosaccharide conformation, and particularly about the function of the periplasmic receptors in transport and chemotaxis.


Effect of transglutaminase treatment on the properties of cast films of soy protein isolates.
The objective of this work was to investigate the effect of microbial transglutaminase (MTGase) treatment on the properties and microstructures of soy protein isolate (SPI) films cast with 0.6 plasticizer per SPI (gg(-1)) of glycerol, sorbitol and 1:1 mixture of glycerol and sorbitol, respectively. Tensile strength (TS), elongation at break (EB), water vapor transmission rate (WVTR) or water vapor permeability (WVP), moisture content (MC), total soluble matter (TSM), lipid barrier property and surface hydrophobicity of control and MTGase-treated films were evaluated after conditioning film specimens at 25 degrees C and 50% relative humidity (RH) for 48 h. The treatment by 4 units per SPI (Ug(-1)) of MTGase increased the TS and surface hydrophobicity by 10-20% and 17-56%, respectively, and simultaneously significantly (P< or =0.05) decreased the E, MC and transparency. The WVTR or TSM of SPI films seemed to be not significantly affected by enzymatic treatment (P>0.05). The MTGase treatment also slowed down the moisture loss rate of film-forming solutions with various plasticizers during the drying process, which was consistent with the increase of surface hydrophobicity of SPI films. Microstructural analyses indicated that the MTGase-treated films of SPI had a rougher surface and more homogeneous or compact cross-section compared to the controls. These results suggested that the MTGase treatment of film-forming solutions of SPI prior to casting could greatly modify the properties and microstructures of SPI films.


In situ measurement of degranulation as a biosensor based on RBL-2H3 mast cells.
A direct degranulation assay has been developed to enable the use of RBL mast cells as a biosensor for screening chemical libraries for drug discovery and environmental toxicity evaluation. Release of beta-hexosaminidase into the extracellular milleu is widely used to characterize cellular components and mechanisms involved in stimulated exocytosis, including those initiated by crosslinking of IgE receptors on mast cells. To adapt this versatile assay for high throughput screening, we developed a direct, in situ method in which beta-hexosaminidase detection is carried out in a single step, convenient for multi-sample processing and thus for biosensor applications. This direct assay is efficient for measuring exocytosis in antigen-stimulated RBL mast cells, detecting antigen concentrations as low as 1 pM. We also demonstrate its utility in detecting inhibition of degranulation by a known pharmacologic inhibitor that blocks Syk tyrosine kinase activity critical for cell activation.


Organization and structure of the human tissue transglutaminase gene.
Analysis of the genomic organization of tissue transglutaminase shows that the gene is 32.5 kilobases, contains 13 exons and 12 introns. Our results show that the sites for the two alternative splicing forms of tissue transglutaminase we reported earlier are located within exons 6 and 10 respectively. The 5'-upstream region of the gene has several potential regulatory promoter elements, and the 3'-exon contains about 50% of the total cDNA size and codes for the C-terminus of the protein. Alignment of deduced tissue transglutaminase amino acid sequence with other transglutaminases showed very similar intron splice positions.


[The activity of the metabolic processes in phagocytosing cells in sarcoidosis patients].
A study of 109 patients with active sarcoidosis revealed an activation of the metabolic processes of phagocytes but with extension and duration of the disease they subside. Apparently, this is related to exhaustion of the functional capacities of the cells. Though, namely, low initial metabolic activity favours spreading and delay of the pathological process.


Cognitive adaptations arising from nonnative experience of sign language in hearing adults.
Three experiments examined spatial transformation abilities in hearing people who acquired sign language in early adulthood. The performance of the nonnative hearing signers was compared with that of hearing people with no knowledge of sign language. The two groups were matched for age and gender. Using an adapted Corsi blocks paradigm, the experimental task simulated spatial relations in sign discourse but offered no opportunity for linguistic coding. Experiment 1 showed that the hearing signers performed significantly better than the nonsigners on a task that entailed 180 degree rotation, which is the canonical spatial relationship in sign language discourse. Experiment 2 found that the signers did not show the typical costs associated with processing rotated stimuli, and Experiment 3 ruled out the possibility that their advantage relied on seen hand movements. We conclude that sign language experience, even when acquired in adulthood by hearing people, can give rise to adaptations in cognitive processes associated with the manipulation of visuospatial information.


Suppressors of cytokine signaling inhibit effector T cell responses during Mycobacterium tuberculosis infection.
Protective immune responses during Mycobacterium tuberculosis (M. tuberculosis) infection are regulated at multiple levels and critically dependent on the balance in the secretion of pro-inflammatory and regulatory cytokines. A key factor that governs this balance at the cellular level is suppressors of cytokine signaling (SOCS). We recently demonstrated that toll-like receptor 2 and dendritic cell (DC)-SIGNR1 differentially regulate SOCS1 expression in DCs during M. tuberculosis infection. This consecutively regulated IL-12 production and determined M. tuberculosis survival. In this study, we characterized the role of SOCS1 in regulating effector responses from CD4(+) and CD8(+) T cells during M. tuberculosis infection. Our data indicate that T cells from M. tuberculosis-infected mice show increased and differential association of SOCS1 with CD3 and CD28, when compared with uninfected mice. While SOCS1 displays increased association with CD3 than CD28 in CD4(+) T cells; SOCS1 is associated more with CD28 than CD3 in CD8(+) T cells. Further, SOCS1 shows increased association with IL-12 and IL-2 receptors in both CD4(+) and CD8(+) T cells from infected mice when compared with naive mice. Silencing SOCS1 in T cells increased signal transduction from T cell receptor (TCR) and CD28 with enhanced activation of key signaling molecules and proliferation. Significantly, SOCS1-silenced T cells mediated enhanced clearance of M. tuberculosis inside macrophages. Finally, adoptive transfer of SOCS1-silenced T cells in M. tuberculosis-infected mice mediated significant reduction in M. tuberculosis loads in spleen. These results exemplify the negative role played by SOCS1 during T cell priming and effector functions during M. tuberculosis infection.


Diverticular disease of the small bowel: a rare cause of the duodenojejunal flexure obstruction (a case report).
The small bowel is the least common site for diverticula in the entire gastrointestinal tract. Chronic upper intestinal obstruction due to diverticula is very rare. We report a case of multiple small bowel diverticula causing mechanical obstruction of the duodenojejunal flexure.


How to avoid compression: a model study of latex sphere grid sections.
The parameters which might play a role in the compression of plastic-embedded objects are studied. The compression is measured on spherical polystyrene latex particles, used as markers in the grid sectioning technique. By changing independently the hardness of the latex particles through a controlled electron irradiation and the hardness of the embedding medium, it is shown that compression is a local event depending only on the mechanical properties of the sectioned object and not on the properties of its surrounding materials. It is demonstrated on one hand that the intrinsic resin compression diminishes as resin hardness increases, and on the other hand that the latex compression can be completely eliminated after a preliminary irradiation by electrons (the electron-induced vulcanization is equivalent to a hardening). It is thus concluded that compression could be greatly reduced or eliminated if objects were sufficiently hardened during their preparation. Several preparation procedures for biological specimens are suggested. For comparison latex has also been irradiated 31/2 days near a 25 000-Ci 60Co gamma source and near the core of an 8-MW nuclear reactor. Neither of these irradiations was sufficient to produce a hardening equivalent to that of the electron irraidation.


Autoimmune hypophysitis: new developments.
Autoimmune hypophysitis, often referred to as lymphocytic hypophysitis, is defined as an inflammatory condition of the pituitary gland of autoimmune etiology that leads to pituitary dysfunction. However, the pathogenesis of autoimmune hypophysitis is still incompletely defined. Although pathogenic autoantibodies in autoimmune hypophysitis have not yet been reported, it has been suggested that several antibodies may be closely related to pathogenesis. Novel clinical entities that are associated with hypophysitis, such as IgG4-related hypophysitis and anti-PIT-1 antibody syndrome, have recently been reported. The findings demonstrate the heterogeneity of the disease and provide important clues for understanding the pathogenesis and definition of hypophysitis, as well as the significance of antipituitary antibodies. This review focuses on new developments in autoimmune hypophysitis.


Methods for analysis of acetyl-CoA synthase applications to bacterial and archaeal systems.
The nickel- and iron-containing enzyme acetyl-CoA synthase (ACS) catalyzes de novo synthesis as well as overall cleavage of acetyl-CoA in acetogens, various other anaerobic bacteria, methanogens, and other archaea. The enzyme contains a unique active site metal cluster, designated the A cluster, that consists of a binuclear Ni-Ni center bridged to an [Fe(4)S(4)] cluster. In bacteria, ACS is tightly associated with CO dehydrogenase to form the bifunctional heterotetrameric enzyme CODH/ACS, whereas in archaea, ACS is a component of the large multienzyme complex acetyl-CoA decarbonylase/synthase (ACDS), which comprises five different subunits that make up the subcomponent proteins ACS, CODH, and a corrinoid enzyme. Characteristic properties of ACS are discussed, and key methods are described for analysis of the enzyme's multiple redox-dependent activities, including overall acetyl-CoA synthesis, acetyltransferase, and an isotopic exchange reaction between the carbonyl group of acetyl-CoA and CO. Systematic measurement of these activities, applied to different ACS protein forms, provides insight into the ACS catalytic mechanism and physiological functions in both CODH/ACS and ACDS systems.


Biomedical research and the law of informed consent.
Awareness of the legal and ethical parameters of nursing practice is a necessity for nurses in all nursing roles, but some of these same legal and ethical considerations take on an increased significance in the area of clinical research. The most predominant legal issue in the research field in terms of both litigation and legal commentary is the issue of informed consent.


Diffusion coefficients for rigid macromolecules with irregular shapes that allow rotational-translational coupling.
We consider six-dimensional diffusion and frictional tensors for a rigid macromolecule immersed in a viscous fluid at low Reynolds number. Our treatment allows for screwlike properties which couple rotational and translational movements. We show that the center of diffusion of a screwlike body can be distinct from its hydrodynamic center of reaction. Symmetry conditions which ensure coincidence are examined. The center of diffusion is found to be the point of a body with the slowest diffusive movements, while rotations about the center of reaction encounter the least average resistance. The macroscopic translational diffusion coefficient is evaluated from a perturbation analysis of the six-dimensional diffusion equation. We show that methodologies which ignore translational-rotational coupling will necessarily underestimate the diffusion rate of screwlike particles. A procedural framework is presented to calculate diffusion coefficients of complicated bodies. As an example we treat a long bent rod.


A clinical and nutritional comparison of biliopancreatic diversion performed with different common and alimentary channel lengths.
This retrospective study compares the clinical and nutritional outcomes of 100 morbidly obese patients who underwent biliopancreatic diversion (BPD) with common (CC) and alimentary channel (AC) length, respectively, 50/250 and 80/200 cm. One hundred patients who received BPD from October 2006 to November 2011 were identified from a database of bariatric procedures performed at the University Hospital of Messina, and the outcomes in terms of weight loss and morbidity were compared. Forty morbidly obese patients underwent BPD with CC 80 cm and AC 200 cm (group 1) and 60 morbidly obese patients underwent BPD with CC 50 cm and AC 250 cm (group 2). A gradual weight loss was observed in both groups during the first 3 years after the operation without any significant difference between the two groups. Two cases of protein malnutrition occurred in the group 2 (3 %) due to poor patient compliance in terms of adequate dietary protein intake. Sideropenic anemia was found in 42 % of obese patients in group 2 versus 22.5 % in group 1 at third-year follow-up despite adequate supplementation (p = 0.047). Diarrhea occurred more frequently with a shorter CC. Lipophilic vitamin deficiencies occurred more frequently with a shorter CC despite adequate oral supplementation. In the medium term, our series showed that shorter CC was associated with no weight loss advantage but with higher morbidity rate, especially in young and fertile women. We recommend a longer CC (80 cm) to be performed especially in this sub-population of obese patients.


Self management, joint protection and exercises in hand osteoarthritis: a randomised controlled trial with cost effectiveness analyses.
There is limited evidence for the clinical and cost effectiveness of occupational therapy (OT) approaches in the management of hand osteoarthritis (OA). Joint protection and hand exercises have been proposed by European guidelines, however the clinical and cost effectiveness of each intervention is unknown.This multicentre two-by-two factorial randomised controlled trial aims to address the following questions:• Is joint protection delivered by an OT more effective in reducing hand pain and disability than no joint protection in people with hand OA in primary care?• Are hand exercises delivered by an OT more effective in reducing hand pain and disability than no hand exercises in people with hand OA in primary care?• Which of the four management approaches explored within the study (leaflet and advice, joint protection, hand exercise, or joint protection and hand exercise combined) provides the most cost-effective use of health care resources Participants aged 50 years and over registered at three general practices in North Staffordshire and Cheshire will be mailed a health survey questionnaire (estimated mailing sample n = 9,500). Those fulfilling the eligibility criteria on the health survey questionnaire will be invited to attend a clinical assessment to assess for the presence of hand or thumb base OA using the ACR criteria. Eligible participants will be randomised to one of four groups: leaflet and advice; joint protection (looking after your joints); hand exercises; or joint protection and hand exercises combined (estimated n = 252). The primary outcome measure will be the OARSI/OMERACT responder criteria combining hand pain and disability (measured using the AUSCAN) and global improvement, 6 months post-randomisation. Secondary outcomes will also be collected for example pain, functional limitation and quality of life. Outcomes will be collected at baseline and 3, 6 and 12 months post-randomisation. The main analysis will be on an intention to treat basis and will assess the clinical and cost effectiveness of joint protection and hand exercises for managing hand OA. The findings will improve the cost-effective evidence based management of hand OA. identifier: ISRCTN33870549.


Birefringent tissue-mimicking phantom for polarization-sensitive optical coherence tomography imaging.
Tissue birefringence is an important parameter to consider when designing realistic, tissue-mimicking phantoms. Options for suitable birefringent materials that can be used to accurately represent tissue scattering are limited. To introduce a method of fabricating birefringent tissue phantoms with a commonly used material-polydimethylsiloxane (PDMS)-for imaging with polarization-sensitive optical coherence tomography (PS-OCT). Stretch-induced birefringence was characterized in PDMS phantoms made with varying curing ratios, and the resulting phantom birefringence values were compared with those of biological tissues. We showed that, with induced birefringence levels up to 2.1 × 10 - 4, PDMS can be used to resemble the birefringence levels in weakly birefringent tissues. We demonstrated the use of PDMS in the development of phantoms to mimic the normal and diseased bladder wall layers, which can be differentiated by their birefringence levels. PDMS allows accurate control of tissue scattering and thickness, and it exhibits controllable birefringent properties. The use of PDMS as a birefringent phantom material can be extended to other birefringence imaging systems beyond PS-OCT and to mimic other organs.


Diffusion tensor imaging tractography study in bipolar disorder patients compared to first-degree relatives and healthy controls.
We aimed to compare white matter structural changes in specific tracts by diffusion tensor imaging (DTI) tractography in patients with bipolar disorder (BD) I, non-ill first-degree relatives (FDR) of the patients, and healthy controls (HC). In a cross-sectional study, we studied right-handed subjects consisting of 16 euthymic BD I patients, 15 FDR, and 15 HC. The anterior thalamic radiation, uncinate fasciculus, corpus callosum, and cingulum bundle were reconstructed by DTI tractography. Mean fractional anisotropy (FA) and apparent diffusion coefficient (ADC) values were compared for group differences followed by post-hoc analysis. The three groups did not differ in terms of sociodemographic variables. There were significant group differences in the FA values among the BD I patients, their FDR, and the HC for the corpus callosum, the dorsal part of the right cingulum bundle, the hippocampal part of the cingulum bundle bilaterally, and the uncinate fasciculus (P < 0.001). The FA values in the patients were significantly lower than in controls, and FDR also showed similar differences; however, they were smaller than those in patients. No significant difference was found between the groups for FA values of the dorsal part of the left cingulum bundle and anterior thalamic radiation. Significant differences were present for ADC values among the groups for the corpus callosum, the dorsal and hippocampal parts of the cingulum, anterior thalamic radiation, and uncinate fasciculus bilaterally (P < 0.01). The FA and ADC values did not correlate significantly with age or any clinical variables. These findings suggest that BD patients and their FDR show alterations in microstructural integrity of white matter tracts, compared to the healthy population.


The Role of Positional Release Therapy in Treating Recalcitrant Brachial Plexus Neuritis: A Case Report.
A 17-year-old female soccer player presented with severe right shoulder pain and scapular winging due to brachial plexus neuritis. The patient was diagnosed with Parsonage-Turner syndrome, a rare condition often resistant to traditional physical therapy, which typically persists for 6 months to years, at times requiring surgical intervention. Over the course of 6 weeks, the patient received positional release therapy once a week coupled with electrical modalities, massage, and a daily home exercise program. This case report is unique because we believe we were the first to use positional release therapy for treatment and the patient's condition resolved more quickly than is typically reported.


Polycomb group proteins are key regulators of keratinocyte function.
The Polycomb group (PcG) proteins are epigenetic suppressors of gene expression that function through modification of histones to change chromatin structure and modulate gene expression and cell behavior. Recent studies show that PcG proteins are expressed in epidermis, that their levels change during differentiation and in disease states, and that PcG expression is regulated by agents that influence cell proliferation and survival. The results indicate that PcG proteins regulate keratinocyte cell-cycle progression, apoptosis, senescence, and differentiation. These proteins are expressed in progenitor cells, in the basal layer, and in suprabasal keratinocytes, and the level, timing, and distribution of expression suggest that the PcG proteins have a central role in maintaining the balance between cell survival and death in multiple epidermal compartments. Additional studies indicate an important role in skin cancer progression.


Release of prostacyclin into the coronary venous blood in patients with coronary arterial disease.
Voluntary patients with a history of myocardial infarction and with typical effort angina underwent catheterization of the coronary sinus and a brachial artery. Healthy young males, serving as controls, were subjected to the same procedure. Arterial and coronary venous blood was drawn at rest and during atrial pacing to angina (patients) or to a heart rate of 140 beats/min (healthy volunteers) for analysis of 6-ketoprostaglandin F1 alpha (6-keto-PGF1 alpha) and prostacyclin-like activity (PILA). 6-Keto-PGF1 alpha levels were measured using radioimmunoassay; PILA in the blood was assayed by rapid preparation of platelet-rich plasma followed by determination of the ADP-induced platelet aggregation. Increased arterial levels of PILA and of radioimmunoactive 6-keto-PGF1 alpha (RIA-6-keto-PGF1 alpha) were observed in the patients at rest as well as during pacing. No obvious release of RIA-6-keto-PGF1 alpha occurred at rest, either in the patients or in the controls. However, during pacing, increased amounts of RIA-6-keto-PGF1 alpha appeared in the coronary venous blood of the patients. The results demonstrate that an increased cardiac prostacyclin formation prevails in patients with signs of impaired coronary flow and suggest that ischemic heart disease is characterized by an insufficient vascular response to this vasodilator prostaglandin rather than by its insufficient endogenous production.


Operational and biochemical aspects of co-digestion (co-AD) from sugarcane vinasse, filter cake, and deacetylation liquor.
This work performed co-AD from the vinasse and filter cake (from 1G ethanol production) and deacetylation liquor (from the pretreatment of sugarcane straw for 2G ethanol production) in a semi-Continuous Stirred Tank Reactor (s-CSTR) aiming to provide optimum operational parameters for continuous CH4 production. Using filter cake as co-substrate may allow the reactor to operate throughout the year, as it is available in the sugarcane off-season, unlike vinasse. A comparison was made from the microbial community of the seed sludge and the reactor sludge when CH4 production stabilized. Lactate, butyrate, and propionate fermentation routes were denoted at the start-up of the s-CSTR, characterizing the acidogenic phase: the oxidation-reduction potential (ORP) values ranged from -800 to -100 mV. Once the methanogenesis was initiated, alkalizing addition was no longer needed as its demand by the microorganisms was supplied by the alkali characteristics of the deacetylation liquor. The gradual increase of the applied organic load rates (OLR) allowed stabilization of the methanogenesis from 3.20 gVS L-1 day-1: the highest CH4 yield (230 mLNCH4 g-1VS) and average organic matter removal efficiency (83% ± 13) was achieved at ORL of 4.16 gVS L-1 day-1. The microbial community changed along with the reactor operation, presenting different metabolic routes mainly due to the used lignocellulosic substrates. Bacteria from the syntrophic acetate oxidation (SAO) process coupled to hydrogenotrophic methanogenesis were predominant (~ 90% Methanoculleus) during the CH4 production stability. The overall results are useful as preliminary drivers in terms of visualizing the co-AD process in a sugarcane biorefinery integrated to scale. KEY POINTS: • Integration of 1G2G sugarcane ethanol biorefinery from co-digestion of its residues. • Biogas production from vinasse, filter cake, and deacetylation liquor in a semi-CSTR. • Lignocellulosic substrates affected the biochemical routes and microbial community. • Biomol confirmed the establishment of the thermophilic community from mesophilic sludge.


Systemic Delivery of CRISPR/Cas9 Targeting HPV Oncogenes Is Effective at Eliminating Established Tumors.
The recent advancements in CRISPR/Cas9 engineering have resulted in the development of more targeted and potentially safer gene therapies. The challenge in the cancer setting is knowing the driver oncogenes responsible, and the translation of these therapies is hindered by effective and safe delivery methods to target organs with minimal systemic toxicities, on-target specificity of gene editing, and demonstrated lack of long-term adverse events. Using a model system based on cervical cancer, which is driven by the ongoing expression of the human papillomavirus E6 and E7 proteins, we show that CRISPR/Cas9 delivered systemically in vivo using PEGylated liposomes results in tumor elimination and complete survival in treated animals. We compared treatment and editing efficiency of two Cas9 variants, wild-type (WT) Cas9 and the highly specific FokI-dCas9, and showed that the latter was not effective. We also explored high-fidelity repair but found that repair was inefficient, occurring in 6%-8% of cells, whereas non-homologous end joining (NHEJ) was highly efficient, occurring in ∼80% of the cells. Finally, we explored the post gene-editing events in tumors and showed that cell death is induced by apoptosis. Overall, our work demonstrates that in vivo CRISPR/Cas editing treatment of preexisting tumors is completely effective despite the large payloads.


Nanodisc scaffold peptide (NSPr) replaces detergent by reconstituting acyl-CoA:cholesterol acyltransferase 1 into peptidiscs.
To conduct biochemical studies in vitro, membrane proteins (MPs) must be solubilized with detergents. While detergents are great tools, they can also inhibit the biological activity and/or perturb oligomerization of individual MPs. Nanodisc scaffold peptide (NSPr), an amphipathic peptide analog of ApoA1, was recently shown to reconstitute detergent solubilized MPs into peptidiscs in vitro. Acyl-coenzyme A:cholesterol acyltransferase 1 (ACAT1), also known as sterol O-acyltransferase 1 (SOAT1), plays a key role in cellular cholesterol storage in various cell types and is a drug target to treat multiple human diseases. ACAT1 contains nine transmembrane domains (TMDs) and primarily forms a homotetramer in vitro and in intact cells; deletion of the N-terminal dimerization domain produces a homodimer with full retention in catalytic activity. ACAT1 is prone to inactivation by numerous detergents. Here we pursued the use of NSPr to overcome the detergent-induced inactivation of ACAT1 by generating near detergent-free ACAT1 peptidiscs. Based on native-PAGE analysis, we showed that NSPr reconstitutes ACAT1 into soluble peptidiscs, in which ACAT1 exists predominantly in oligomeric states greater than a homotetramer. The formation of these higher-order oligomeric states was independent of the N-terminal dimerization domain, suggesting that the oligomerization is mediated through hydrophobic interactions of multiple ACAT1 subunits. ACAT1 peptidiscs were still susceptible to heat-mediated inactivation, presumably due to the residual detergent (CHAPS) bound to ACAT1. We then conditioned ACAT1 with phosphatidylcholine (PC) to replace CHAPS prior to the formation of ACAT1 peptidiscs. The results showed, when PC was included, ACAT1 was present mainly in higher-order oligomeric states with greater enzymatic activity. With PC present, the enzymatic activity of ACAT1 peptidiscs was protected from heat-mediated inactivation. These results support the use of NSPr to create a near detergent-free solution of ACAT1 in peptidiscs for various in vitro studies. Our current results also raise the possibility that, under certain conditions, ACAT1 may form higher-order oligomeric states in vivo.


Secretory vesicle swelling by atomic force microscopy.
The swelling of secretory vesicles has been implicated in exocytosis, but the underlying mechanism of vesicle swelling remained unknown. Earlier studies from our laboratory demonstrated the association of the alpha-subunit of heterotrimeric GTP-binding protein G(alphai3) with zymogen granule membrane and implicated its involvement in vesicle swelling. Mas7, an active mastoparan analog known to stimulate Gi proteins, was found to stimulate the GTPase activity of isolated zymogen granules and cause swelling. Increase in vesicle size in the presence of GTP, NaF, and Mas7 were irreversible and found to be KCl sensitive. However, Ca2+ had no effect on zymogen granule size. Taken together, these results indicated that zymogen granules, the membrane-bound secretory vesicles in exocrine pancreas, swell in response to GTP mediated by a G(alphai3) protein. Subsequently, our studies demonstrated that the water channel aquaporin-1 (AQP1) is also present at the zymogen granule membrane and participates in rapid GTP-induced and G(alphai3)-mediated vesicular water gating and swelling. Isolated zymogen granules exhibit low basal water permeability. However, exposure of granules to GTP results in a marked potentiation of water entry. Treatment of zymogen granules with the known water channel inhibitor Hg2+ is accompanied by a reversible loss in both the basal and GTP-stimulable water entry and vesicle swelling. Introduction of AQP1-specific antibody raised against the carboxy-terminal domain of AQP1 blocked GTP-stimulable swelling of vesicles. Our results demonstrate that AQPI associated at the zymogen granule membrane is involved in basal GTP-induced and G(alphai3)-mediated rapid gating of water into zymogen granules of the exocrine pancreas.


Corrigendum: Earlywood and Latewood Stable Carbon and Oxygen Isotope Variations in Two Pine Species in Southwestern China during the Recent Decades.
[This corrects the article on p. 2050 in vol. 7, PMID: 28119725.].


Effect of feeding a high-fat diet during pregnancy on glucose metabolism in the rat.
To alter glucose homeostasis in a period of great glucose demand, pregnant rats were submitted to a high-fat diet and compared to virgin rats. In virgin rats, blood glucose, ketone bodies, plasma insulin, and free fatty acids were not affected by the diet consumed. Glucose turnover measured in the postabsorptive period was slightly decreased in virgin rats fed a high-fat diet compared to rats fed a standard diet. Assuming that the glucose turnover rate is representative for the 24-hour average endogenous glucose production, in rats fed a standard diet the daily carbohydrate intake (9.2 +/- 0.7 g/d) exceeded the glucose turnover rate (4 +/- 0.2 g/d) and could meet the glucose requirement. In rats fed a high-fat diet the carbohydrate intake (2.7 +/- 0.2 g/d) was lower than the glucose turnover rate (3.8 +/- 0.2 g/d), which demonstrated the need for an active endogenous glucose production. Blood glucose, ketone bodies, plasma insulin, and free fatty acid concentrations followed the same patterns during pregnancy in rats fed a standard diet compared to rats fed a high-fat diet. The glucose turnover rate in the postabsorptive period was no more decreased by the high-fat diet in pregnant rats compared to virgin rats despite the greater glucose demand. In late pregnancy the glucose turnover rate was increased up to 70%.(ABSTRACT TRUNCATED AT 250 WORDS)


Prevalence and factors associated with condom use among Chinese monogamous female patients with sexually transmitted infection in Hong Kong.
Monogamous female sexually transmitted infection (STI) patients are at high risk of recurrent STI, but there is no study investigating their risk behaviors. We investigated the prevalence of male condom use and associated factors among monogamous STI female patients. Independent variables included socio-demographic characteristics, STI history, nature of relationship, and cognitions (Health Belief Model). Dependent variables included the use of male condoms in the past two months and intention to use male condoms consistently in the future 6 months. A cross-sectional survey using a structured questionnaire was conducted among STI female patients with only one male sex partner in the last 12 months and attended a major government STI clinic in Hong Kong. Among the 538 participants, 25.1% used condoms consistently (every time) during sex in the past 2 months, while 35.2% intended to do so in the next 6 months. A multiple stepwise logistic regression model identified three statistically significant factors associated with lower likelihoods of consistent condom use in the past 2 months: type of sole sex partner (cohabitant vs. husband: OR = 0.29, 95% CI = 0.12 to 0.70; regular boyfriend vs. husband: OR = 0.52, 95% CI = 0.30 to 0.91), being financially dependent (OR = 0.45, 95% CI = 0.27 to 0.75), and partner's dislike of condom use (OR = 0.23, 95% CI = 0.15 to 0.39). In a similar model, the same three factors were found to be significantly associated with the intention to use condoms consistently in the next 6 months. The low prevalence of intention to use condoms consistently in the future suggested that our study population might be victimized again. The impact of financially dependent relationships and men's dislike of using condoms on condom use may operate through imbalanced gender power. Interventions are greatly warranted and should increase risk awareness and empower this vulnerable population.


High rates of COVID-19 vaccine refusal among Afghan pregnant women: a cross sectional study.
Severe Acute Respiratory Syndrome Corona Virus Type-2 (SARS-COV-2) was first detected in China and created a global pandemic rapidly. Subsequently after serious efforts different types of vaccines produced against the virus and recommended for all people including pregnant women. The aim of this study was to realize the willingness of pregnant women to accept the COVID-19 vaccine in Kabul Afghanistan. For this purpose, a cross-sectional study was conducted in gynecology wards of several hospitals in Kabul, Afghanistan from 10th of July to 20th of August 2021 through direct interview with the pregnant women who had come for prenatal care to the healthcare centers. The collected data were analyzed through Statistical Package for Social Studies (SPSS) version 25. Simple descriptive analysis computed for demographic characteristics and bi-variable (Chi-square) analysis was carried out to find out the associations of taking vaccine with demographic variables. A p-value of < 0.05 was considered significant at 95% confidence interval. A total of 491 who were completed the inclusion criteria included in the analysis. A small portion of pregnant women (8.6%) illustrated the intent to get the COVID-19 vaccine if it is recommended by the health authorities in Afghanistan. Our study found a high rate of COVID-19 vaccine refusal among pregnant women in Kabul, Afghanistan. They showed the concern on adverse effects of the vaccine as the main reason for refusal, emphasizing the need to reduce the misconception on vaccine efficacy and campaigns to enhance awareness on the vaccine safety and benefits for mothers and babies.


Expression Profiles of Immune Cells after Propofol or Sevoflurane Anesthesia for Colorectal Cancer Surgery: A Prospective Double-blind Randomized Trial.
The antitumor effects of natural killer cells, helper T cells, and cytotoxic T cells after cancer surgery were reported previously. This study hypothesized that propofol-based anesthesia would have fewer harmful effects on immune cells than volatile anesthetics-based anesthesia during colorectal cancer surgery. In total, 153 patients undergoing colorectal cancer surgery were randomized and included in the analysis. The primary outcome was the fraction of circulating natural killer cells over time in the propofol and sevoflurane groups. The fractions of circulating natural killer, type 1, type 17 helper T cells, and cytotoxic T cells were investigated. The fractions of CD39 and CD73 expressions on circulating regulatory T cells were investigated, along with the proportions of circulating neutrophils, lymphocytes, and monocytes. The fraction of circulating natural killer cells was not significantly different between the propofol and sevoflurane groups until 24 h postoperatively (20.4 ± 13.4% vs. 20.8 ± 11.3%, 17.9 ± 12.7% vs. 20.7 ± 11.9%, and 18.6 ± 11.6% vs. 21.3 ± 10.8% before anesthesia and after 1 and 24 h after anesthesia, respectively; difference [95% CI], -0.3 [-4.3 to 3.6], -2.8 [-6.8 to 1.1], and -2.6 [-6.2 to 1.0]; P = 0.863, P = 0.136, and P = 0.151 before anesthesia and after 1 and 24 h, respectively). The fractions of circulating type 1 and type 17 helper T cells, cytotoxic T cells, and CD39+ and CD73+ circulating regulatory T cells were not significantly different between the two groups. The neutrophil to lymphocyte ratio in both groups remained within the normal range and was not different between the groups. Propofol-based anesthesia was not superior to sevoflurane-based anesthesia in terms of alleviating suppression of immune cells including natural killer cells and T lymphocytes during colorectal cancer surgery.


Cancer surveillance strategies in ulcerative colitis: the need for modernization.
The risk of colorectal cancer is increased in patients with long-standing ulcerative colitis. Traditional surveillance has centered around regular standard white-light colonoscopy, with multiple biopsies aimed at detecting dysplasia or the identification of early cancer. This has resulted in only a modest reduction in cancer incidence and mortality. A better understanding of disease risk factors may allow endoscopic resources to be more focused on patients at higher risk. In addition, advanced endoscopic techniques have the potential to improve dysplasia detection, minimize the need for routine biopsies, and allow for the removal of dysplastic lesions, avoiding the need for surgery. Techniques such as magnification colonoscopy, chromoendoscopy, narrow band imaging, autofluorescence, and confocal endomicroscopy may all have a role to play in improving the benefits of endoscopic surveillance. Revised endoscopic surveillance strategies are proposed, incorporating aspects of risk stratification, a well-established practice in noncolitis-related colorectal cancer screening, and some of these new technologies.


Speciation and evaluation of Arsenic in surface water and groundwater samples: a multivariate case study.
The principal object of the current study was to estimate total arsenic and its inorganic speciation in different origins of surface water (n=480) and groundwater (n=240) of Sindh, Pakistan. This study provided a description based on the evaluation of physico-chemical parameters of collected water samples and possible distribution of As with respect to its speciation. The concentration of total inorganic As (iAs) and its species (As(3+) and As(5+)) for the surface and underground water was reported in terms of basic statistical parameters, principal component analysis, cluster analysis, metal-to-metal correlations and linear regression analyses. The chemical correlations were observed by PCA, which were used to classify the samples by CA, based on the PCA scores. Standard addition method confirmed the accuracy; the recoveries of As(3+) and iAs were found to be >98%. The concentration of As(5+) in the water samples was calculated by the difference of the total inorganic arsenic and As(3+). The results revealed that the groundwater of the understudied area was more contaminated as compared to the surface water samples. The mean concentration of As(3+) and As(5+) in the surface water and groundwater samples were in the range 3.0 to 18.3 and 8.74-352 microg/L, respectively.


Quantification of Bax/Bcl-2 ratios in peripheral blood lymphocytes, monocytes and granulocytes and their relation to susceptibility to anti-Fas (anti-CD95)-induced apoptosis.
Neutrophils have the shortest half-life among circulating leucocytes and rapidly undergo apoptosis in vitro. The homologous Bcl-2 and Bax proteins have opposing effects, with Bcl-2 extending cellular survival and Bax promoting cell death following an apoptotic stimulus. We determined Bcl-2 to Bax expression ratios in peripheral blood lymphocytes, monocytes and granulocytes and related them to the susceptibility of these cells to anti-Fas (anti-CD95)-induced apoptosis. Here, we show that Bax/Bcl-2 ratios are high in granulocytes and relatively low in monocytes and lymphocytes. Furthermore, we show a relation between this ratio in the different leucocyte subsets and their susceptibility to anti-Fas-induced apoptosis, with granulocytes showing the highest susceptibility, followed by monocytes and lymphocytes. It is concluded that the balance between Bcl-2 and Bax forms an apoptotic rheostat, which seems to determine sensitivity to apoptosis.


Accurate measurement of stable isotopes 46Ca and 48Ca in human feces, plasma, and urine in relation to human nutrition of calcium.
A method based on Radiochemical Neutron Activation Analysis (RNAA) is described which allows simultaneous measurement of two stable isotopes of calcium, 46Ca and 48Ca, in human feces, plasma, and urine for the purpose of studying human nutrition and metabolism of calcium. It is shown that these measurements can be made with relative analytical precision of 1--5% depending on the particulars of a given experiment. The method has been applied in humans and data are given showing that kinetics of plasma appearance of 46 Ca administered orally with food can be readily investigated. This method allows investigation of a number of important nutritional and metabolic issues in all human population groups without regard to radioisotope safety considerations, and should prove especially helpful in relation to studies of calcium bioavailability from different foods in a variety of population groups for whom use of radiocalcium is not warranted.


Hormonal receptors in locally advanced breast cancer: change with response to neoadjuvant chemotherapy?
Twenty-one cancers in 20 patients with locally advanced breast cancer were studied. Incisional biopsy was performed without using electrocautery. Tissue was obtained for histology and estrogen (ER) and progesterone (PR) receptors. Patients received chemotherapy after biopsy. All responded and had radical mastectomy performed. Tissue was removed from the mastectomy specimen to confirm residual tumor and for repeat receptor measurements. Initial receptor levels were negative in 13 cancers. Following chemotherapy, both ER and PR levels were unchanged in 11 cancers. Levels in one cancer changed from ER-PR- to ER+PR- and one changed from ER-PR- to ER+PR+. Six cancers were ER+PR+ at initial examination. Repeat receptor levels after chemotherapy were ER+PR+ in three. One ER+PR+ tumor became ER-PR+, one changed to ER+PR-, and one to ER-PR-. One ER+PR- cancer remained ER+PR- after treatment and an ER-PR+ cancer became ER-PR- after treatment. Chemotherapy dose not significantly alter hormonal levels in breast cancer tissue.


Dendritic cell-based immunotherapy.
Dendritic cell (DC)-based vaccinations represent a promising approach for the immunotherapy of cancer and infectious diseases as DCs play an essential role in initiating cellular immune responses. A number of clinical trials using ex vivo-generated DCs have been performed so far and only minor toxicity has been reported. Both the induction of antigen-specific T cells and clinical responses have been observed in vaccinated cancer patients. Nevertheless, DC-based immunotherapy is still in its infancy and there are many issues to be addressed such as antigen loading procedures, DC source and maturational state, migration properties, route, frequency, and dosage of DC vaccination. The increasing knowledge of DC biology should be used to improve the efficacy of this new therapy.


Battling the COVID-19 infodemic in an Irish context: the role of iHealthFacts.
On the 11 th of March 2020, the World Health Organisation (WHO) declared a global pandemic due to the SARS-CoV-2 virus, which causes coronavirus disease 2019 (COVID-19). This was one month after Dr. Tedros Adhanom Ghebreyesus, Director-General of the WHO declared that we are also fighting an 'infodemic'. The WHO has described an infodemic as an "over-abundance of information - some accurate and some not - that makes it hard for people to find trustworthy sources and reliable guidance when they need it". iHealthFacts.ie is an Irish resource where the public can quickly and easily check the credibility and reliability of health claims circulating on social media. Unreliable claims can lead to poorly informed health choices. iHealthFacts is an initiative that supports the public to think critically about health claims and make well-informed choices. Here, we describe the role iHealthFacts plays in providing reliable information to the public and offer reflections from those involved in launching this initiative during a pandemic.


Identification of hub genes and immune infiltration in ulcerative colitis using bioinformatics.
Ulcerative colitis (UC) is a chronic inflammatory disease of the intestine, whose pathogenesis is not fully understood. Given that immune infiltration plays a key role in UC progression, our study aimed to assess the level of immune cells in UC intestinal mucosal tissues and identify potential immune-related genes. The GSE65114 UC dataset was downloaded from the Gene Expression Omnibus database. Differentially expressed genes (DEGs) between healthy and UC tissues were identified using the "limma" package in R, while their Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were determined with the clusterProfiler package. Protein-protein interaction network analysis and visualization were performed with STRING and Cytoscape. Immune cell infiltration was calculated with CIBERSORT. The relationship between hub genes and immune-infiltrated cells in UC was determined by Pearson correlation. A total of 206 DEGs were identified, of which 174 were upregulated and 32 downregulated. GO and KEGG functional classification indicated DEG enrichment in immune response pathways, including Toll-like receptor signaling, IL-17 signaling, and immune system process and chemokine signaling. 13 hub genes were identified. Infiltration matrix analysis of immune cells showed abundant plasma cells, memory B cells, resting CD4 memory T cells, γδ T cells, M0 and M1 macrophages, and neutrophils in UC intestinal tissues. Correlation analysis revealed 13 hub genes associated with immune-infiltrated cells in UC. 13 hub genes associated with immune-infiltrated cells in UC were identified; they included CXCL13, CXCL10, CXCL9, CXCL8, CCL19, CTLA4, CCR1, CD69, CD163, IL7R, PECAM1, TLR8 and TLR2. These genes could potentially serve as markers for the diagnosis and treatment of UC.


Engraftment of mucosal stem cells into murine jejunum is dependent on optimal dose of cells.
Transplantation of intestinal mucosal stem cells is an important step in the development of intestinal gene therapy and treatment of intestinal mucosal diseases. We hypothesized that engraftment rates increase proportionally with increasing doses of seeded stem cells and increasing jejunal débridment. Intestinal mucosal organoids were harvested from neonatal mice carrying a green-fluorescent protein (GFP) transgene and transplanted into adult GFP(-) mice (n = 66). In recipients, two jejunal segments (1.5 cm) were isolated with their blood circulation left intact with anastomosis of the distal and proximal segments to restore continuity. Debridement of native enterocytes was performed by perfusing luminally with ethylene diamine tetraacetc acid solutions for 20 min. A total of 5,000, 10,000, or 25,000 organoids were then seeded. Three weeks later, cross sections (n = 398) of the segments were evaluated for the presence of GFP(+) neomucosa using fluorescence microscopy. Additional segments were débrided for 30 and 40 min (n = 83). Other conditions were not tested because of the rate of high mortality in these experiments. The group seeded with 10,000 organoid units at 20 min showed the highest engraftment of GFP(+) epithelium. Engraftment was improved by increasing débridment times at this seeding density. Overall mortality was 70%. These findings suggest that there is an optimal seeding density of stem cell clusters for enhanced engraftment in this model. Mortality prohibited complete testing of all combinations of seeding density and debridement times.


Pathologic correlation with near infrared-indocyanine green guided surgery for pediatric liver cancer.
Hepatoblastoma (HB) and hepatocellular carcinoma (HCC) are the most common primary malignant tumors of childhood. Intraoperative indocyanine green (ICG) administration with near-infrared imaging (NIR) has emerged as a surgical technology that can be used to assist with localization of pulmonary metastases secondary to HB; however, there has been limited application as an adjunct for resection of the primary liver tumor and assessment of extrahepatic disease. We present 14 patients treated for HB, HCC, and malignant rhabdoid tumor at our institution with the use of intraoperative NIR-ICG guidance. All patients were treated with 0.2-0.75 mg/kg IV ICG, 48-96 h prior to surgery. Intraoperative NIR-ICG guided imaging was performed with several commercial devices. Intraoperative NIR-ICG guidance allowed pulmonary metastasectomy in five patients using thoracoscopy or thoracotomy allowing for visualization of multiple nodules not seen on preoperative imaging most of which were positive for malignancy. NIR-ICG guidance allowed for assessment of extrahepatic extension in three patients; an HCC patient with extrahepatic lymph node extension of disease, an HB patient with extrapulmonary thoracic recurrence in the diaphragm and chest wall, and a patient with tumor rupture at diagnosis with peritoneal nodules at the time of surgery. This technique was used to guide partial hepatectomy in 11 patients for which the technique enabled successful identification of tumor and tumor margins. Three patients had nonspecific staining of the liver secondary to decreased timing from ICG injection to surgery or biliary obstruction. NIR-ICG enabled resection of satellite HB lesions in three multifocal patients and confirmed a benign satellite lesion in two additional patients. Intraoperative use of NIR-ICG imaging during partial hepatectomy enabled enhanced identification and guidance for surgical resection of extrahepatic disease and multifocal liver tumors for the treatment of children with primary liver cancer.