α-ketoglutarate suppresses immediate early gene expression in cancer cells.
Cancer cells exhibit increased glutamine consumption compared to normal cells, supporting cell survival and proliferation. Glutamine is converted to α-ketoglutarate (αKG), which then enters the tricarboxylic acid cycle to generate ATP. Recently, therapeutic modulation of glutamine metabolism has become an attractive metabolic anti-cancer strategy. However, how synergistic combination therapy is required to overcome glutamine metabolism drug resistance remains elusive. To address this issue, we first investigated the role of αKG in regulating gene expression in several cancer cell lines. Using RNA-seq analysis and histone modification screening, we demonstrated that αKG reduced the expression of the immediate early gene (IEG) in cancer cells in an H3K27 acetylation-dependent manner. Conversely, glutaminase (GLS) inhibitors induce IEG expression in cancer cells. Furthermore, we showed that siRNA knockdown of orphan nuclear receptor subfamily 4 group A member 1 (NR4A1) induces IEG expression. Notably, the NR4A1 agonist cytosporone B sensitizes GLS inhibitor resistance to cancer cell death. Together, these findings indicate that therapeutic targeting of IEG dysregulation by αKG can be a potentially effective anti-cancer therapeutic strategy for glutamine metabolism inhibitors.


Self-evaluation of automated vehicles based on physics, state-of-the-art motion prediction and user experience.
Legal restrictions allow to give full control to automated vehicles for longer time periods either in restricted areas or when moving with reduced speed. Although being technically feasible for a wide range of driving scenarios, the restrictions are still in place due to the lack of a clear safety strategy. An essential step towards safety is the introduction of a self-monitoring component. In this study, a self-evaluation concept is presented which assesses a system based on a physics-defined minimum prediction horizon for state-of-the-art Deep Learning-based trajectory prediction models. Since User Experience is a key metric for car manufacturers, a further manoeuvre constraint is added to the model. We emphasize the generalizability of the presented assessment concept, however, in order to demonstrate feasibility in practical use, three specific scenarios are discussed. The results are gained with real data from publicly available driving campaigns as well as synthetically generated simulation data. Two exemplary models, a simple LSTM-based model and VectorNet, a prominent motion prediction model, are evaluated. A quantitative assessment shows a lack of training data in the public datasets for vehicle speeds > 25 m/s in order to offer safe driving above such vehicle speeds.


ROS production and gene expression in alveolar macrophages exposed to PM(2.5) from Baghdad, Iraq: Seasonal trends and impact of chemical composition.
The objective of this study was to assess the impact of changes in atmospheric particulate matter (PM) composition on oxidative stress markers in an in-vitro alveolar macrophage (AM) model. Fifty-three PM2.5 samples were collected during a year-long PM sampling campaign in Baghdad, Iraq, a semi-arid region of the country. Monthly composites were analyzed for chemical composition and for biological activity using in-vitro measurements of ROS production and gene expression in the AM model. Twelve genes that were differentially expressed upon PM exposure were identified and their co-associations with the composition of PM2.5 were examined. Ten of those genes were up-regulated in January and April composites; samples which also exhibited high ROS activity and relatively high PM mass concentration. ROS production was statistically correlated with total PM2.5 mass, levoglucosan (a wood burning tracer) and several trace elements of the PM (especially V and Ni, which are associated with oil combustion). The expression of several cytokine genes was found to be moderately associated with PM mass, crustal materials (indication of dusty days or dust storms) and certain metals (e.g. V, Fe and Ni) in the PM. Thus, the ROS activity association with PM2.5, may, in part, be driven by redox-active metals. The antioxidant response genes (Nqo1 and Hmox1) were moderately associated with polyaromatic hydrocarbons (PAHs) and showed a good correlation (r-Pearson of >0.7) with metals linked to vehicle-related emissions (i.e. Cu, Zn and Sb). Examining these associations in a larger sample pool (e.g. daily samples) would improve the power of the analysis and may strengthen the implication of these chemicals in the oxidative stress of biological systems, which could aid in the development of new metrics of PM toxicity.


HVJ-liposome mediated gene transfer into hepatocytes in vivo.
The efficient transduction of appropriate target cells will be critical for gene therapy. We evaluated the suitability of hemagglutinating virus of Japan (HVJ)-liposome-mediated gene transfer for gene therapy of liver diseases. The Escherichia coli beta-galactosidase (beta-gal) gene was introduced into rat liver by HVJ-liposome to examine gene transfer efficacy and persistence of expression with or without partial hepatectomy prior to transfection. About 30% of hepatocytes were transduced after portal vein injection. Gene expression was transient, with only 2% of hepatocytes expressing beta-gal after 4 weeks. However, partial hepatectomy performed 24 h prior to injection resulted in persistently high levels of beta-gal for 4 weeks after injection. A 247-bp beta-gal polymerase chain reaction fragment transcript was detected in livers of transfected rats, but not in livers of control rats. The rat livers following gene transfer were histologically normal, and serum glutamic-pyruvic transaminase was not found to be elevated in rats. Our results demonstrate that HVJ-liposome-mediated gene transfer produced high gene transduction and persistent gene expression in the liver.


Mitogenic activity in platelet-poor plasma from rats with persistent liver nodules or liver cancer.
Platelet-poor plasma (PPP) from F-344 rats with chemically-induced preneoplastic liver nodules or hepatocellular carcinoma stimulated S-phase DNA synthesis in monolayer cultures of normal rat hepatocytes. Similar mitogenic activity was detected in PPP 6 hrs to 1 week after partial hepatectomy (PH) or after necrotizing doses of CCl4 or diethylnitrosamine (DENA). Very little activity was found in PPP4 from control rats. The mitogenic activity in PPP from animals with nodules was non-dialyzable (greater than 14 kd) and bound to a heparin-sepharose affinity column. None of the mitogenic PPPs competed with [125I] epidermal growth factor (EGF) for binding sites on A431 cells or normal rat hepatocytes. These studies indicate that persistent proliferation of preneoplastic and neoplastic hepatocytes is associated with increased circulating levels of mitogenic hepatocyte growth factor.


Ultrahigh performance liquid chromatography-triple quadrupole mass spectrometry inhibitors fishing assay: a novel method for simultaneously screening of xanthine oxidase inhibitor and superoxide anion scavenger in a single analysis.
Xanthine oxidase (XOD) inhibitors and superoxide anion scavengers play an important role in the treatment of gout and the inhibition of many diseases related to superoxide anion. The respective quantitation of uric acid and superoxide anion by traditional spectroscopic methods is routine in XOD inhibitors and superoxide anion scavengers screening at laboratories worldwide. In the present study, we established an ultrahigh performance liquid chromatography and triple quadrupole mass spectrometry (UHPLC-TQ-MS) method of higher accuracy and speed that combines screening of superoxide anion scavenger and XOD inhibitor in a single analysis by adding WST-1 (2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium sodium salt) to the enzymatic reaction. We applied the established method to determine the XOD inhibitory activities and superoxide scavenging activities of some herbal extracts and compounds from natural products, which could be classified into six groups based on the results of the assay. Our innovative protocol is fast, accurate and robust. Moreover, it can eliminate false positive and false negative results which may occur in the traditional spectroscopic methods.


Using tyrosinase as a monophenol monooxygenase: A combined strategy for effective inhibition of melanin formation.
Tyrosinase is a binuclear copper-containing metalloprotein that leads the fast and regio-selective o-hydroxylation of monophenols to o-diphenols. However, the subsequent second oxidation to produce o-quinones, i.e., melanin precursors, from the o-diphenols has restricted its use to the production of functional o-diphenol derivatives. Herein, we present a combined strategy for the effective inhibition of melanin formation in tyrosinase reaction, which allows the use of tyrosinase as a monophenol monooxygenase. The o-diphenolic products were protected from being oxidized in the tyrosinase reaction by borate ions and L-ascorbic acid (LAA). Borate-o-diphenol complexes were favorable formed at high pH and consequentially protected the o-diphenolic products from the catecholase activity of tyrosinase. LAA not only directly reduced the byproduct, o-quinones, into o-diphenols but also assisted the completion of the tyrosinase reaction cycle by removing a hydroxyl group attached to the copper metal cluster at the active site of the met-form tyrosinase. The regio-selective o-hydroxylation of 7,4'-dihydroxyisoflavone (daidzein) to produce 7,3',4'-trihydroxyisoflavone (3'-ODI) was successfully carried out by whole E. coli cell biotransformation with heterologously expressed tyrosinase from Bacillus megaterium. The yield of this o-hydroxylation of 5 mM daidzein in one-pot 400 mL reaction was ca. 100% in 90 min and the productivity was 16.3 mg 3'-ODI · L(-1) · h(-1) · DCW mg(-1), which is considerably higher than that of other monooxygenases. The method effectively abolished melanin synthesis, so that the o-diphenolic product remained stable without enzyme inactivation. Other monophenolic phytochemicals such as resveratrol and genistein could be subjected to the same strategy. After 1 h, 1 mM of genistein and resveratrol were both converted to orobol and piceatannol, respectively, with ca. 95% conversion yield. These results support the strong potential of tyrosinase as a monooxygenase for regio-selective o-hydroxylation of various monophenolic compounds.


Heart transplantation under cyclosporine or tacrolimus combined with mycophenolate mofetil or everolimus.
In this study, we examined whether cyclosporine was effective when combined with everolimus in clinical heart transplantation (HT). From August 2004 to July 2007, 108 adult patients underwent primary HT. The main exclusion criteria were: donors > 60 years; cold ischemia times > 6 hours; recipients of multiorgan transplantation or a previous transplantation; and panel-reactive antibodies > or = 25%. The cyclosporine plus everolimus regimen (group CE, n = 32) was suggested first; upon refusal or if the recipient or donor was positive for hepatitis B surface antigen or PCR + hepatitis C infection, then patient was randomly assigned to success cyclosporine plus mycophenolate mofetil (MMF; group CM, n = 24) or tacrolimus plus MMF (group TM, n = 25). All patients underwent similar operative procedures and postoperative care with protocol endomyocardial biopsies. No 30-day mortality was noted in any group. The efficacy failure rates were 3%, 25%, and 16% in groups CE, CM, and TM, respectively (P = .04 between groups CE and CM). The 1-year survivals were 96.7% +/- 18.1%, 89.7% +/- 29.8%, and 81.0% +/- 35.5% for groups CE, CM, and TM, respectively (P = .04 between groups CE and TM). The 3-year survival rates were 91.9% +/- 28.3%, 79.8% +/- 46.0%, and 81.0% +/- 35.5% in groups CE, CM, and TM, respectively. The 3 immunosuppressive regimens offered good efficacy after HT. The cyclosporine plus everolimus regimen showed a significantly better result with less efficacy failure (compared with cyclosporine plus MMF: 3% vs 25%) and better 1-year survival compared with tacrolimus plus MMF: 96.7% vs 81.0%.


Panum's fusional area estimated with a criterion-free technique.
It has been reported that criterion-free estimates of the upper disparity limits for fusion of line targets are small enough to be accounted for by monocular vernier sensitivity. However, targets such as lines, which contain high spatial frequencies, may ensure small fusion limits, since fusion limits obtained with criterion-dependent methods for narrow-band targets, such as sinusoids or difference-of-Gaussian luminance profiles, are proportional to target spatial periods. Experiment 1 therefore explored whether criterion-free methods give fusion limits for narrow-band targets that can be accounted for by vernier sensitivity. Vertical fusion limits were estimated by a method that forced observers to discriminate a disparate sinusoidal grating from an immediately adjacent zero-disparity grating. Fusion limits were too large to be explained by monocular vernier thresholds obtained for the same targets. In addition, fusion limits were not affected by large changes in target contrast, whereas vernier thresholds increased as contrast was decreased. The results of Experiment 1 also argued against interocular suppression as the cause of single vision, since vernier offsets that were visible when viewed monocularly were invisible under binocular viewing conditions. In Experiment 2, manual adjustment of disparities yielded fusion limits little different from those obtained with the forced-choice method of Experiment 1, demonstrating that it is possible to design adjustment methods for assessing fusion limits that are as sensitive as forced-choice methods. In Experiment 3, large reductions in target contrast, which have the effect of decreasing disparity sensitivity, did not alter fusion limits, disconfirming the idea that fusion limits estimated with discriminative procedures represent disparity-detection thresholds. In Experiment 4, disparities were adjusted until a just noticeable difference in grating contrast appeared. These disparities were larger than fusion limits, indicating that fusion limits did not represent a change in apparent contrast arising from disparity limitations of binocular summation. Together, the four experiments support the existence of binocular fusion as a unique category of sensory performance, disconfirm several nonfusional explanations of single vision, and support the use of criterion-free as well as adjustment methods in measuring fusion limits.


Stiffness characteristics analysis of a Biglide industrial parallel robot considering the gravity of mobile platform and links.
For the machining process of industrial parallel robots, the gravity generated by the weight of mobile platform and links will lead to the deviation of the expected machining trajectory of the tool head. In order to evaluate this deviation and then circumvent it, it is necessary to perform the robotic stiffness model. However, the influence of gravity is seldom considered in the previous stiffness analysis. This paper presents an effective stiffness modeling method for industrial parallel robots considering the link/joint compliance, the mobile platform/link gravity, and the mass center position of each link. First, the external gravity corresponding to each component is determined by the static model under the influence of gravity and mass center position. Then, the corresponding Jacobian matrix of each component is obtained by the kinematic model. Subsequently, the compliance of each component is obtained by cantilever beam theory and FEA-based virtual experiments. In turn, the stiffness model of the whole parallel robot is determined and the Cartesian stiffness matrix of the parallel robot is calculated at several positions. Moreover, the principal stiffness distribution of the tool head in each direction over the main workspace is predicted. Finally, the validity of the stiffness model with gravity is experimentally proved by the comparison of the calculated stiffness and measured stiffness in identical conditions.


Cellular distribution of okadaic acid in the digestive gland of Mytilus galloprovincialis (Lamarck, 1819).
The distribution of okadaic acid between the digestive and the secretory cells of the digestive gland of Mytilus galloprovincialis was studied for the main purpose of determining if they might be responsible for the two-compartment depuration kinetics found in previous studies. The two cell types did not accumulate okadaic acid to the same degree. However, the concentrations found in each cellular type were not consistent with those expected from the output of the two-compartment depuration model, suggesting that a mechanism other that the differential accumulation in cellular types is involved. Binding to some yet undetermined cellular components is suggested.


Measurement of temporal lobe T2 relaxation times using a routine diagnostic MR imaging protocol in epilepsy.
To determine the applicability of a fast spin-echo (FSE) pulse sequence for T2 relaxation time measurements in diagnostic imaging of temporal lobe epilepsy (TLE) and in epilepsy research. To compare FSE T2-relaxometry to the measurements with multi-echo sequence and visual assessment of MR scans. MR imaging and T2 relaxometry was performed with widely used 1.5 T scanner only. Fast dual-echo sequence (TE-14/85 ms) and multi-echo pulse sequence were used for T2 measurements. Normal ranges of T2 values in regions of interest in temporal lobe were estimated in 20 healthy controls. Sixty-five patients with intractable focal epilepsy were studied. Fifty-five patients had TLE, three multilobar focal epilepsy and seven extratemporal focal epilepsy. T2 measurements with the FSE showed good reproducibility in the test objects and control subjects. In one TLE case unilateral focal T2 changes were not identified visually. T2-relaxometry was more sensitive than visual inspection of MR scans in assessing bilateral hippocampal changes: there were 15 cases with abnormal bilateral T2 values. Visually bilateral changes were detected in six out of these 15 cases (40%). In six cases (40%) only unilateral changes were diagnosed visually, and in three cases (20%) bilateral changes were classified as probable with qualitative evaluation. T2 relaxation time measurement supplied additional objective data in cases with ambiguous hippocampal changes on visual assessment: T2-relaxometry confirmed hippocampal abnormalities in seven cases judged visually as probable. In four cases with the suspicion of hippocampal changes T2 values appeared to be normal. In TLE patients, images constructed from FSE sequences can be used to estimate T2 relaxation times easily and reliably. T2 measurements are an objective method to diagnose structural changes in the temporal lobe. T2-relaxometry is most helpful to assess bilateral hippocampal abnormalities, and thus might have an impact on estimating postsurgical outcome.


Meaning of illness and quality of life in patients with type 2 diabetes.
Investigations into personal factors influencing quality of life are important for those developing strategies to support patients with diabetes. This study aimed to investigate the influence of meaning of illness on quality of life in patients with type 2 diabetes. Veterans from primary care clinics in the southeastern United States completed a questionnaire including questions from the validated 5-scale Meaning of Illness Questionnaire (MIQ). Unadjusted and adjusted linear regression models investigated the physical and mental components of quality of life with the 5 MIQ factors. The sample comprised 302 Black and White veterans. The physical component of quality of life (PCS) was positively associated with type of stress/attitude of harm (β = 2.43, CI: 0.94 to 3.93) and challenge/motivation/hope (β = 3.02, CI: 0.40 to 5.64) after adjustment, whereas the mental component of quality of life (MCS) was positively associated with the degree of stress/change in commitment (β = 2.58, CI: 0.78 to 4.38), and negatively associated with challenge/motivation/hope (β = -2.55, CI: -4.99 to -0.11). Attitudes of challenge, motivation and hope had opposite effects on mental and physical components of quality of life in this sample of veterans. Additionally, whereas, the type of stress and attitude towards harm or loss was associated with the physical component, the degree of stress and change in commitments was associated with the mental component. This suggests addressing the meaning of an illness may be complex but is an important consideration in improving both physical and mental components of quality of life in patients with type 2 diabetes.


First-in-Human Case Study: Pregnancy in Women With Crohn's Perianal Fistula Treated With Adipose-Derived Stem Cells: A Safety Study.
The aim of this study was to determine whether treatment with adipose-derived stem cells (ASCs) had any influence on fertility, course of pregnancy, newborn weight, or physical condition of newborns. We performed a retrospective study of patients with a desire to become pregnant after having received intralesional injection of autologous ASCs for the treatment of perianal or rectovaginal fistula associated with Crohn's disease. We collected data on the resulting pregnancies, deliveries, and newborns of these patients. ASCs were expanded in vitro and characterized according to the international guidelines for cell surface markers (clusters of differentiation) and differentiated to adipocytes, chondrocytes, and osteocytes prior to implantation (except first implant in 2002). We analyzed five young women with Crohn's disease treated with ASCs: one for rectovaginal and perianal fistula, two for rectovaginal fistula only, and two for perianal fistula only. All patients received 2 doses of 20 million and 40 million cells at an interval of 3-4 months. Another patient received 2 doses of 6.6 million and 20 million ASCs with 9 months between each dose. Fertility and pregnancy outcomes were not affected by cell therapy treatment. No signs of treatment-related malformations were observed in the neonates by their respective pediatricians. In the patients studied, cell therapy with ASCs did not affect the course of pregnancy or newborn development. Local treatment with mesenchymal stem cells derived from adipose tissue seems not to affect the ability to conceive, the course of pregnancy, pregnancy outcomes, or newborns' health in female patients. This is the first publication about pregnancy outcome in women with perianal fistula and Crohn's disease treated with stem cell therapy, and could be of interest for doctors working in cell therapy. This is a very important question for patients, and there was no answer for them until now.


Proteinuria and its consequences in renal disease.
Chronic renal diseases that involve proteinuria are typically characterized by an inexorable progression to end-stage renal failure. Many studies suggest that this progression may be the result of factors, such as intraglomerular hypertension and glomerular hypertrophy, that are unrelated to the initial disease. This paper reviews the mechanisms of progression of chronic renal diseases and discusses therapeutic strategies that should prevent or minimize further renal damage and the applicability of these strategies to patients with the rare X-linked lysosomal storage disorder Fabry disease. Renal involvement is a major feature of Fabry disease, which is characterized by vacuolated epithelial cells in the glomerulus and distal tubules, resulting from lipid inclusions within these cells. Although enzyme replacement therapy is the key strategy to halt the progression of Fabry disease, additional therapeutic options include blood pressure control, reduction of proteinuria, lipid control and inhibition of the renin-angiotensin system. A range of therapeutic options, used in conjunction with enzyme replacement therapy, may have beneficial effects on the renal manifestations of Fabry disease.


Torrents of torment: turbulence as a mechanism of pulsatile tinnitus secondary to venous stenosis revealed by high-fidelity computational fluid dynamics.
Pulsatile tinnitus (PT) is a debilitating condition that can be caused by a vascular abnormality, such as an arterial or venous lesion. Although treatment of PT-related venous lesions has been shown to successfully cure patients of the associated 'tormenting' rhythmical sound, much controversy still exists regarding their role in the etiology of PT. A patient presented with a history of worsening, unilateral PT. A partial venous sinus obstruction related to the large arachnoid granulation was detected on the right side, and subsequently stented at the right transverse sinus. High-fidelity computational fluid dynamics (CFD) was performed on a 3D model digitally segmented from the pre-stent venogram, with assumed pulsatile flow rates. A post-stent CFD model was also constructed from this. Data-driven sonification was performed on the CFD velocity data, blinded to the patient's self-reported sounds. The patient reported that the PT was completely resolved after stenting, and has had no recurrence of the symptoms after more than 2 years. CFD simulation revealed highly disturbed, turbulent-like flow at the sigmoid sinus close to auditory structures, producing a sonified audio signal that reproduced the subjective sonance of the patient's PT. No turbulence or sounds were evident at the stenosis, or anywhere in the post-stent model. For the first time, turbulence generated distal to a venous stenosis is shown to be a cause of PT. High-fidelity CFD may be useful for identifying patients with such 'torrents' of flow, to help guide treatment decision-making.


Elevated symptoms of executive dysfunction predict lower adaptive functioning in 3-year-olds with autism spectrum disorder.
Executive functioning (EF) deficits co-occur frequently with autism spectrum disorder (ASD) and have a long-term detrimental impact on quality of life of children and their families. Timely identification of risk for EF vulnerabilities may hasten access to early intervention and alleviate their long-term consequences. This study examines (1) if EF deficits are elevated in toddlers with ASD compared to nonautistic siblings of children with ASD, typically developing (TYP) toddlers, and toddlers with atypical developmental presentation; and (2) if EF deficits have a detrimental effect on adaptive functioning in ASD. Participants were recruited between September 2014 and October 2019 and included 73 toddlers with ASD, 33 nonautistic siblings of children with ASD, 35 toddlers with atypical development, and 28 TYP toddlers matched on chronological age (M = 39.01 months, SD = 3.11). EF deficits were measured using the BRIEF-P; adaptive skills were measured using the VABS-II. Whenever appropriate, analyses were controlled for MSEL verbal and nonverbal developmental quotient, ADOS-2 autism severity scores, and sex. Analyses revealed that toddlers with ASD exhibited elevated BRIEF-P scores across all domains compared to each of the three comparison groups. Higher BRIEF-P scores were associated with lower adaptive social, communication, and daily living skills while controlling for symptom severity, verbal and nonverbal functioning, and sex. In conclusion, marked vulnerabilities in EF are already present in 3-year-old toddlers with ASD and are predictive of the level of adaptive functioning in ASD. EF vulnerabilities in toddlers should be targeted for intervention to improve long-term outcomes in ASD. LAY SUMMARY: Many children with autism experience vulnerabilities in executive functioning (EF), which may include challenges with inhibition, working memory, cognitive flexibility, and planning. The study shows that these vulnerabilities can already be detected at age three and that their presence is linked with lower social, communication, and daily living skills. Screening children with ASD for EF challenges and helping those who have difficulties may improve their long-term outcomes.


Determinants of maternal immunization in developing countries.
Maternal immunization is an effective intervention to protect newborns and young infants from infections when their immune response is immature. Tetanus toxoid vaccination of pregnant women is the most widely implemented maternal vaccine in developing countries where neonatal mortality is the highest. We identified barriers to maternal tetanus vaccination in developing African and Asian countries to identify means of improving maternal immunization platforms in these countries. We categorized barriers into health system, health care provider and patient barriers to maternal tetanus immunization and conducted a literature review on each category. Due to limited literature from Africa, we conducted a pilot survey of health care providers in Malawi on barriers they experience in immunizing pregnant women. The major barriers of the health system are due to inadequate financial and human resources which translate to inadequate vaccination services delivery and logistics management. Health care providers are limited by poor attendance of Antenatal Care and inadequate knowledge on vaccinating pregnant women. Patient barriers are due to lack of education and knowledge on pregnancy immunization and socioeconomic factors such as low income and high parity. There are several factors that affect maternal tetanus immunization. Increasing knowledge in health care providers and patients, increasing antenatal care attendance and outreach activities will aid the uptake of maternal immunization. Health system barriers are more difficult to address requiring an improvement of overall immunization services. Further analyses of maternal immunization specific barriers and the means of addressing them are required to strengthen the existing program and provide a more efficient delivery system for additional maternal vaccines.


Mechanism of alveolar bone destruction in periodontitis - Periodontal bacteria and inflammation.
Periodontal disease is an inflammatory disease caused by periodontopathogenic bacteria, which eventually leads to bone tissue (alveolar bone) destruction as inflammation persists. Periodontal tissues have an immune system against the invasion of these bacteria, however, due to the persistent infection by periodontopathogenic bacteria, the host innate and acquired immunity is impaired, and tissue destruction, including bone tissue destruction, occurs. Osteoclasts are essential for bone destruction. Osteoclast progenitor cells derived from hematopoietic stem cells differentiate into osteoclasts. In addition, bone loss occurs when bone resorption by osteoclasts exceeds bone formation by osteoblasts. In inflammatory bone disease, inflammatory cytokines act on osteoblasts and receptor activator of nuclear factor-κB ligand (RANKL)-producing cells, resulting in osteoclast differentiation and activation. In addition to this mechanism, pathogenic factors of periodontal bacteria and mechanical stress activate osteoclasts and destruct alveolar bone in periodontitis. In this review, we focused on the mechanism of osteoclast activation in periodontitis and provide an overview based on the latest findings.


Host cell reactivation of cisplatin-damaged plasmid DNA in human non-T leukocyte cell lines.
Host-cell reactivation of cisplatin-damaged pRSVcat was assessed in three B cell lines (SKW 6.4, WIL2-NS, RPMI 1788), the monocytic cell line THP-1, and promyelocytic HL-60 cells. IC50 values following a 3-day exposure of the five leukocyte cell lines to cisplatin ranged from 0.45 to 1.92 microM. Transfer of pRSVcat into all cell lines was effected by electroporation and the resultant CAT activity was measured 24 h later by a rapid single vial CAT assay. CAT activity corresponding to an average of 0.06 units of purified CAT enzyme was expressed by WIL2-NS cells. Very low to no expression of the CAT vector was observed in all other cell lines studied, despite the presence of intracellular levels of 3H-labelled pRSVcat comparable to WIL2-NS. Epstein-Barr virus transformed B cells (SKW 6.4 and RPMI 1788) did not successfully perform host cell reactivation. In WIL2-NS cells, platination of pRSVcat to defined levels of 5-40 platinum molecules per plasmid led to a graded reduction in CAT activity expressed following transfection. Platination levels of 20 and 40 platinum molecules per plasmid did not alter the efficiency of transfer of pRSVcat into these cells by electroporation. Data obtained in this study suggests that EBV transformation may possibly be a negative influence on host cell-reactivation assays for cisplatin-DNA damaged plasmid in non-T human leukocytes.


Molecular detection and typing of human papillomavirus in laryngeal carcinoma specimens.
Human papillomavirus (HPV) DNA was detected in 32% of laryngeal carcinoma biopsy samples studied. The genotypes identified were high-risk types, the most frequent being HPV 16. Viral DNA was integrated into the host genome (genotype HPV 16), providing supporting evidence for a role of HPV in the carcinogenic pathway of laryngeal squamous cell carcinoma. HPV has been detected in laryngeal lesions, both benign and neoplastic, with a variable frequency (8-60%). These viral agents have been proposed as an adjuvant or cofactor in head and neck carcinogenesis because of their oncogenic properties. The aims of this study were to identify HPV in laryngeal carcinoma samples and to describe the physical state of the viral genome, i.e. its integration to the host DNA. Formalin-fixed, paraffin wax-embedded tumor samples from patients with newly diagnosed laryngeal carcinomas were collected. The HPV genome was identified using polymerase chain reaction (PCR) with primers complementary to the conserved region L1 (MY09-11). Genotyping was accomplished by restriction fragment length polymorphism. Samples positive for HPV 16 were assayed by PCR with primers complementary to region E2, interrupted during viral genome integration. Ten of the 31 samples (32%) were positive for HPV DNA and all of the samples were positive for human beta-globin. The genotypes identified were HPV 16 (n=3), HPV 58 (n=2) and HPV 39, 45, 51, 59, 66 and 69 (n=1 for each). The three samples positive for HPV 16 had lost region E2, meaning that the viral DNA had been integrated into the host genome.


Tissue-specific regulation of lipogenic mRNAs by thyroid hormone.
We have previously shown that triiodothyronine (T3) regulates rat fatty acid synthesis in a tissue specific manner. Here, we determined the effects of thyroid state on mRNAs encoding the lipogenic enzymes, acetyl CoA carboxylase (ACC) and fatty acid synthase (FAS). S14 mRNA, a sequence tightly associated with lipogenesis, was also measured. Levels of the three mRNA were 9-13-fold higher in hyper- than hypothyroid liver. Limited expression in kidney and heart was also increased by thyroid hormone. In brown adipose tissue, highest levels were recorded in hypothyroid animals. Thyroid state did not affect expression in lung and brain. All these changes are consistent with those previously measured in fatty acid synthesis. In white adipose tissue, mRNA expression was increased by hyperthyroidism. This increase may not be reflected in fatty acid synthesis, since we recently showed lipogenesis to be reduced under these circumstances. All three mRNAs responded rapidly to T3 in liver, but more slowly in kidney and fat. Thus, T3 regulates lipogenesis by altering levels of ACC and FAS mRNAs. S14 mRNA changes in parallel.


Abstract of the 7th Annual Scientific Meeting, International Academy of Pathology, Malaysian Division Kuantan 2021: Dermatopathology, Uropathology and Gynaecologic pathology.
No abstract available.


The integrated comprehension of lncRNA HOXA-AS3 implication on human diseases.
Long non-coding RNA (lncRNA) is a non-protein-coding RNA with a length of more than 200 nucleotides. Studies have shown that lncRNAs have vital impacts on various pathological processes and participate in the development of human diseases, usually through acting as competing endogenous RNAs to modulate miRNA expression and biological functions. lncRNA HOXA Cluster Antisense RNA 3 (HOXA-AS3) was a newly discovered lncRNA and has been demonstrated to be abnormally expressed in many diseases. Moreover, HOXA-AS3 expression was closely correlated with the clinicopathologic characteristics in cancer patients. In addition, HOXA-AS3 exhibited significant properties in regulating several biological processes, including cell proliferation, invasion, and migration. Furthermore, HOXA-AS3 has provided promising values in the diagnosis, prognosis, and therapeutic strategies of several diseases such as liver cancer, glioma, lung cancer, oral cancer, gastric cancer, and even atherosclerosis. In this review, we discuss the abnormal expression of HOXA-AS3 in several human disorders and some pathobiological processes and its clinical characteristics, followed by a summary of HOXA-AS3 functions, regulatory mechanisms, and clinical application potential.


Role of cytokines in altitude-associated preeclampsia.
BACKGROUND: Preeclampsia (PE) is more common at high than low altitude and contributes to the altitude-related decline in birth weight. Since inflammatory markers are implicated in PE, we asked if such markers differed in PE vs. normotensive pregnant (NORM) women residing at high altitude (3600-4100 m), and were related to uterine artery blood flow (UA BF) or fetal growth. METHODS: Subjects were 33 Andean pregnant residents of Bolivia, comprising six with early-onset PE (≤ 34 wk), 12 with late-onset PE (> 34 wk), and 15 gestational-age matched NORM. Maternal pro- and anti-inflammatory cytokines were measured using a multiplex bead-based assay and UA BF by Doppler ultrasound. RESULTS: PE compared to NORM women had higher levels of the pro-inflammatory cytokines IL-6 and IL-8 as well as higher levels of the anti-inflammatory cytokine IL-1ra, but only IL-6 levels were higher when gestational age was controlled. Women with early- vs. late-onset PE had higher TNFα levels, and higher IL-6 was negatively correlated with birth weight in all women at ≤ 34 wk. We suggest that pro-inflammatory factors influence both the timing and severity of PE at high altitude.


Immunoglobulin-specific T-B cell interaction. IV. B cell presentation of idiotypic determinant(s) of monoclonal anti-surface immunoglobulin antibody to idiotope-recognizing helper T clones.
Previously, we have demonstrated T-B cell interactions mediated by T cell recognition of immunoglobulin (Ig) peptide/major histocompatibility complex (MHC) class II complexes derived by the B cell processing of endogenously synthesized Ig molecules. In this report Ig-specific T-B cell interaction mediated by B cell presentation of idiotopes (Id) of anti-sIg antibodies to Id-specific T cell clones has been studied in Ig kappa-1-congenic rat strains. A panel of August (RT-1c; Ig kappa-1a) rat T helper clones specific for Id of syngeneic anti-Ig kappa-1b C3B9 monoclonal antibody (mAb) has been developed to study IdC3B9 presentation by Ig kappa-1b-bearing B cells from congenic August.1b (RT-1c; Ig kappa-1b) rats. Five of seven IdC3B9-specific T clones responded even at very low concentrations (100-200 pg/ml) of C3B9 mAb presented by Ig kappa-1b+ B cells. In contrast, the presentation of intact C3B9 mAb by nonspecific antigen-presenting cells (macrophages, Ig kappa-1a+ B cells, etc.) to IdC3B9-specific T cells was of low efficiency. The IdC3B9-specific T cell response to idiotopes of anti-Ig kappa-1b C3B9 mAb was found to be restricted by RT-1B molecule and required the processing of intact C3B9 molecule. IdC3B9 epitope recognized by C31 and C5 clones was mapped to the heavy chain of C3B9 mAb. Thus, B cells are able to present peptides related to the V region of anti-sIg Ab, i.e. Id peptide/MHC class II complexes, to Id-recognizing T cells. IdC3B9-presenting B cells are specifically activated both to proliferation and Ig production upon interaction with IdC3B9-specific T clones. Based on the results of our studies on B cell presentation of Ig epitopes to T cells a hypothetical model of Ig peptide-driven T-B cell interaction has been proposed.


Dilated cardiomyopathy-associated BAG3 mutations impair Z-disc assembly and enhance sensitivity to apoptosis in cardiomyocytes.
Dilated cardiomyopathy (DCM) is characterized by dilation of left ventricular cavity with systolic dysfunction. Clinical symptom of DCM is heart failure, often associated with cardiac sudden death. About 20-35% of DCM patients have apparent family histories and it has been revealed that mutations in genes for sarcomere proteins cause DCM. However, the disease-causing mutations can be found only in about 17% of Japanese patients with familial DCM. Bcl-2-associated athanogene 3 (BAG3) is a co-chaperone protein with antiapoptotic function, which localizes at Z-disc in the striated muscles. Recently, BAG3 gene mutations in DCM patients were reported, but the functional abnormalities caused by the mutations are not fully unraveled. In this study, we analyzed 72 Japanese familial DCM patients for mutations in BAG3 and found two mutations, p.Arg218Trp and p.Leu462Pro, in two cases of adult-onset DCM without skeletal myopathy, which were absent from 400 control subjects. Functional studies at the cellular level revealed that the DCM-associated BAG3 mutations impaired the Z-disc assembly and increased the sensitivities to stress-induced apoptosis. These observations suggested that BAG3 mutations present in 2.8% of Japanese familial DCM patients caused DCM possibly by interfering with Z-disc assembly and inducing apoptotic cell death under the metabolic stress.


Preoperative naproxen sodium reduces postoperative pain following arthroscopic knee surgery.
This study was undertaken to assess the efficacy of a single preoperative dose of naproxen sodium in reducing postoperative pain and length of day surgery stay in patients undergoing arthroscopic knee surgery. A randomized, double-blind clinical trial was carried out on 66 ASA I and ASA II patients scheduled for arthroscopic knee surgery. The treatment group (n = 26) received two capsules containing 275 mg of naproxen sodium each, and the control group (n = 40) received placebo. Preoperative and postoperative visual analogue pain scores, postoperative analgesic requirements in hospital as well as 24 hr after discharge, and length of day surgery stay were studied. There was a decrease in postoperative pain, both in hospital (naproxen 0.7 +/- 1.2 vs placebo 2.2 +/- 2.3) and at 24 hr after discharge (naproxen 0.8 +/- 1.9 vs placebo 3.8 +/- 3.2) (P = 0.0001). There was no difference in the need for in-hospital postoperative analgesics or in the time to discharge. However, there was a difference in the use of analgesics after discharge (naproxen group 30.4% vs placebo group 71.4%) (P < 0.01). The results of this study suggest that a single preoperative dose of 550 mg naproxen sodium is effective in reducing postoperative pain in arthroscopic knee surgery, both in the immediate postoperative period and for up to 24 hr after the completion of surgery.


Microorganisms and their metabolites affect the content of polycyclic aromatic hydrocarbons and their derivatives in pyrolyzed material.
Toxic polycyclic aromatic hydrocarbons (PAHs) and more toxic N- and O-containing derivatives can be determined in biochar. However, their fate in the environment and bioavailability depends on many parameters and was not studied yet. In the presented studies a set of biochars obtained from various feedstock at the same pyrolysis temperature (600 °C) subjected to environmental pressure e.g. soil microorganisms and enzymes was described. Presented study aimed to determine the effect of biological agents on the physicochemical characteristic and the content of PAHs and their derivatives in biochars after long-term treatment (6 months). The results indicated that enzymatic aging usually lowered (up to 94 %) the content of PAHs and their derivatives in biochar. Simultaneously, biological aging reduced the bioavailability of tested compounds. Considering the total fraction of PAHs and their derivatives, biochars treated with nutrients and microbial inoculum were characterized by the lowest content of analytes (even in comparison to biochars treated with nutrients alone). To complement the obtained results, the content of C, H, N, O, and ash as well as specific surface area, aromaticity, polarity, and hydrophilicity in biochar before and after modifications were determined. In general, enzymatic aging increased, and biological aging decreased the content of C% and H% in biochar. Both aging processes lowered the H/C ratio which indicated the decrease of the aromatization degree for artificially altered biochar.


Association between ultra-processed foods intake with lipid profile: a cross-sectional study.
The purpose of this cross-sectional study was to examine the association between ultra-processed foods (UPFs) intake and lipid profile in Iranian people. The study was performed on 236 individuals with the age range of 20-50 years in Shiraz, Iran. Food intakes of the participants were evaluated using a 168-item food frequency questionnaire (FFQ) which was previously validated in Iranian populations. In order to estimate the ultra-processed foods intake, classification of NOVA food group was used. Serum lipids including total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) were measured. The results showed that mean of age and body mass index (BMI) of the participants were 45.98 years and 28.28 kg/m2, respectively. Logistic regression was used to evaluation the relation between UPFs intake and lipid profile. Higher UPFs intake was associated with increased OR of TG and HDL abnormality in both crude (OR 3.41; 95% CI 1.58, 7.34; P-trend = 0.001 and OR 2.99; 95% CI 1.31, 6.82; P-trend = 0.010) and adjusted models (OR 3.69; 95% CI 1.67, 8.16; P-trend = 0.001 and OR 3.38 95% CI 1.42, 8.07; P-trend = 0.009). But, there were no association between UPFs intake and other indices of lipid profile. Also, we found significant associations between UPFs intake and dietary nutrient profiles. In conclusion, UPFs consumption could worsen the nutritional profile of the diet and lead to negative changes in some indices of the lipid profile.


Narrow-band imaging with dual focus magnification in differentiating colorectal neoplasia.
Real-time optical diagnosis of colorectal polyps may lead to substantial time and cost savings and could potentially reduce complications associated with polypectomy. We prospectively assessed the utility of a novel narrow-band imaging (NBI) system with dual focus magnification (DF) in differentiating colorectal polyps in consecutive patients undergoing colonoscopy. All procedures were carried out with a prototype 190 series Exera III NBI system with Dual Focus capability. Histology of each polyp was predicted in real time with NBI-DF based on the modified Sano's classification with a confidence level (low/high). NBI-DF diagnosis was then compared to the final (blinded) histopathology results. Primary endpoint was the accuracy of endoscopic prediction with the modified Sano's classification of all polyps when they were diagnosed with high confidence. Secondary endpoints were the accuracy of post-polypectomy surveillance intervals for diminutive polyps(≤ 5 mm). A total of 164 polyps in 87 patients (53 males) with a mean age of 63 (range 28-86) years were evaluated. 149 polyps were diagnosed with high confidence on endoscopic prediction, out of which 80 were diminutive, 31 small (6-9 mm) and 38 large (>10 mm). Overall accuracy of NBI-DF compared to final histopathology was 97%. The accuracy for post-polypectomy surveillance interval based on the endoscopic prediction was 97%. The NPV for diminutive rectosigmoid polyps for adenomatous histology was 100%. In this preliminary feasibility study, NBI-DF permitted prediction of histology of colorectal polyps with high accuracy. In addition, both of the secondary endpoints exceeded the ASGE PIVI thresholds for the management of diminutive polyps.


Protein structural changes in keratin fibers induced by chemical modification using 2-iminothiolane hydrochloride: a Raman spectroscopic investigation.
For the purpose of investigating in detail the influence of chemical modification using 2-iminothiolane hydrochloride (2-IT) on keratin fibers, the structure of cross-sections at various depths of white human hair, treated with 2-IT and then oxidized, was directly analyzed without isolating the cuticle and cortex, using Raman spectroscopy. In particular, the beta-sheet and/or random coil content (beta/R) and the alpha-helix (alpha) content in human hair fibers were estimated by amide I band analysis. The S-S band intensity, amide III (unordered) band intensity, and beta/R content existing from the cuticle region to the center of cortex region of virgin white human hair remarkably increased by performing the chemical modification using 2-IT. On the other hand, not only the S-S band intensity, but also S-O band intensity existing throughout the cortex region of the bleached (damaged) white human hair increased by performing chemical modification using 2-IT. In particular, beta/R content existing throughout the cortex region of the bleached white human hair decreased, while the skeletal C-C stretch (alpha) band intensity at 935 cm(-1) and the alpha content remarkably increased. This indicates a secondary structural change from the random coil form to the alpha-helix form in the proteins existing throughout the cortex region. From these experiments, we concluded that the formation of new disulfide (-SS-) groups resulting from chemical modification using 2-IT induced the secondary structural changes of proteins existing throughout the cortex region.


Treatment effects of R-appliance and Fränkel-2 in Class II division 1 malocclusions.
The purpose of this study was to compare the effects of a differently designed functional appliance (the R- appliance) with Fränkel-2. Twenty-seven patients (16 girls and 11 boys) with a mean age of 9.8 (SD 1.6) years were treated with the R-appliance for 15.4 (SD 0.4) months and twentyseven (15 girls and 12 boys) patients with a mean age of 9.1 (SD 1.1) years were treated with a Fränkel-2 appliance for 19 (SD 5.6) months. All patients had Class II division 1 malocclusions due to mandibular deficiency and all of them had prepubertal stages of skeletal development. Lateral cephalograms obtained at the beginning (T1) and at the end (T2) of the study were analysed. Paired t-tests showed that SNB significantly increased in both groups. The incisor mandibular plane angle (IMPA) was reduced in the R- appliance group by 2.2 (SD 4.9) degrees (P<0.03) but increased by 2.2 (SD 2.6) degrees (P<0.001) in the Fränkel-2 group. The SNA in the R-appliance group showed an increase of 0.2 (SD 2) degrees (P<0.6), while it was decreased by 0.4 (SD 0.5) degrees (P<0.6) in the Fränkel-2 group. Both treatment modalities were successful in moving the mandible forward. However, with the R-appliance, this was achieved without proclination of the lower incisors.


Clinical Experience and Evaluation of Patient Treatment Verification With a Transit Dosimeter.
To prospectively evaluate a protocol for transit dosimetry on a patient population undergoing intensity modulated radiation therapy (IMRT) and to assess the issues in clinical implementation of electronic portal imaging devices (EPIDs) for treatment verification. Fifty-eight patients were enrolled in the study. Amorphous silicon EPIDs were calibrated for dose and used to acquire images of delivered fields. Measured EPID dose maps were back-projected using the planning computed tomographic (CT) images to calculate dose at prespecified points within the patient and compared with treatment planning system dose offline using point dose difference and point γ analysis. The deviation of the results was used to inform future action levels. Two hundred twenty-five transit images were analyzed, composed of breast, prostate, and head and neck IMRT fields. Patient measurements demonstrated the potential of the dose verification protocol to model dose well under complex conditions: 83.8% of all delivered beams achieved the initial set tolerance level of ΔD of 0 ± 5 cGy or %ΔD of 0% ± 5%. Importantly, the protocol was also sensitive to anatomic changes and spotted that 3 patients from 20 measured prostate patients had undergone anatomic change in comparison with the planning CT. Patient data suggested an EPID-reconstructed versus treatment planning system dose difference action level of 0% ± 7% for breast fields. Asymmetric action levels were more appropriate for inversed IMRT fields, using absolute dose difference (-2 ± 5 cGy) or summed field percentage dose difference (-6% ± 7%). The in vivo dose verification method was easy to use and simple to implement, and it could detect patient anatomic changes that impacted dose delivery. The system required no extra dose to the patient or treatment time delay and so could be used throughout the course of treatment to identify and limit systematic and random errors in dose delivery for patient groups.


Split-face comparison of the picosecond 1064-nm Nd:YAG laser using a microlens array and the quasi-long-pulsed 1064-nm Nd:YAG laser for treatment of photoaging facial wrinkles and pores in Asians.
Skin photoaging manifests deeper wrinkles and larger pore size. Various lasers have been tried for rejuvenation of photoaging skin, and the quasi-long-pulsed 1064-nm neodymium:yttrium-aluminum-garnet (Nd:YAG) laser is one promising treatment option. New types of laser device are emerging, including devices operating with picosecond pulse duration combined with a microlens array, which are regarded as a new breakthrough for skin rejuvenation. We aimed to evaluate the clinical effectiveness and safety of the picosecond 1064-nm Nd:YAG laser using a microlens array compared with the quasi-long-pulsed 1064-nm Nd:YAG laser in a split-face design. We designed a split-faced, prospective study and enrolled 25 subjects with photoaging facial wrinkles and enlarged pores. Each facial area was randomly assigned to undergo treatment with either the picosecond 1064-nm Nd:YAG laser (Pico-arm) or the quasi-long-pulsed 1064-nm Nd:YAG laser (Quasi-arm). We performed five laser sessions at 2-week intervals, and final results were assessed after 20 weeks after the initial laser session. We used a five-point global assessment scale, wrinkle and pore index derived from 3D camera analysis. We enrolled a total of 25 subjects (24 females and 1 male) with Fitzpatrick skin types III to IV and a mean age of 63.8 ± 5.7 years. After treatment, 54.2% of subjects in the Pico-arm reported at least moderate improvement in visible pores compared with 41.7% of the Quasi-arm (P = 0.001). A total of 12.5% of subjects in the Pico-arm showed moderate improvement in wrinkles versus 4.2% of the Quasi-arm (P = 0.125). There was a 41.3% reduction in the pore index in the Pico-arm compared to a 3.9% increase in the Quasi-arm (P = 0.048). There was a 16.4% reduction in the wrinkle index in the Pico-arm compared with a 0.5% reduction in the Quasi-arm (P = 0.01). Pain assessment score was higher in the Pico-arm than the Quasi-arm (3.65 ± 1.70 vs 1.28 ± 1.28, P = 0.001). No serious adverse events occurred during the study. Our findings suggest that the picosecond 1064-nm Nd:YAG laser with a microlens array is as effective as the quasi-long-pulsed 1064-nm Nd:YAG laser for treatment of photoaging wrinkles and pores.


Organising haematoma mimicking tumour on MRI following resection of acoustic neuroma.
We describe a 26-year-old woman in whom an enhancing, intradural extramedullary mass was found at the craniocervical junction on MRI 3 years after resection of a large acoustic neuroma. The radiological appearances suggested a new tumour, raising the possibility of neurofibromatosis 2, provoking a review of family members in an attempt to confirm the diagnosis, as well as further surgery. Histology of the lesion revealed an organising haematoma, with no evidence of malignancy. The imaging features can be explained by the process by which blood clot in the subarachnoid space is resorbed. Caution is advised in interpreting CT OR MRI after neurosurgery.


Buffalo (Bubalus bubalis) epiphyseal proteins give protection from arsenic and fluoride-induced adverse changes in acetylcholinesterase activity in rats.
The objective of this study was to determine the effect of fluoride (F) and arsenic (As) on the activity of acetylcholinesterase (AChE), a critically important nervous system enzyme, and to test the protective role of buffalo epiphyseal (pineal) proteins (BEP) in rats. Arsenic (20 mg/kg BW, intraperitoneally) and F (150 ppm, perorally) were exposed, and BEP was administered intraperitoneally (100 μ g/kg BW) along with F and As to rats for 7 days. As and F exposure significantly (p < 0.05) increased their levels in plasma and decreased the activity of AChE in plasma, RBCs, heart, and brain of rats. Interestingly, As- and F-induced inhibition of AChE activities increased As and F levels in plasma, and organs were significantly (p < 0.05) counteracted by BEP administration. These findings indicate the protective role of buffalo (Bubalus bubalis) epiphyseal proteins on F- and As-induced adverse changes in AChE activity as a candidate biomarker for neurotoxicity in female rats.


Cryopreservation of newly formed hybridomas.
A new freezing technique is described which permits time-consuming protocols as a first screening of newly formed hybridomas. In this procedure complete 96 well clustertrays with growing hybridomas are cryopreserved after programmed freezing. This procedure has been successfully applied to a number of fusion protocols for which the objective was to obtain monoclonal antibodies against tissue specific antigens. To this end hybridoma supernatants were screened by an immunoperoxidase technique on frozen sections. Freezing of the hybridoma containing clustertrays permitted extensive screening and partial characterization of the previously collected supernatants. After subsequent thawing of appropriate wells, the hybridoma clones proved to be viable and usually no loss of antibody production was observed.


In vitro antileukaemic activity of extracts from berry plant leaves against sensitive and multidrug resistant HL60 cells.
The aim of the present study was to determine in vitro antileukaemic activity of extracts obtained from selected berry plant leaves (Fragaria x ananassa Duch. cv Elsanta, raspberry Rubus ideus L. cv Polana and blueberry Vaccinium corymbosum L. cv Bluecrop) against promyelocytic HL60 cell line and its multidrug resistant sublines exhibiting two different MDR phenotypes: HL60/VINC (overexpressing P-glycoprotein) and HL60/DOX (overexpressing MRP1 protein). It was found that the blueberry extract was the most efficient against sensitive HL60 cell line (about 2-fold more active than strawberry and raspberry extracts) but presented much lower activity towards resistant cells. In contrast, strawberry and raspberry extracts exhibited the high cytotoxic activity against sensitive leukaemia HL60 cell line as well as its MDR sublines. The values of resistance factor (RF) found for these extracts were very low lying in the range 0.32/2.0.


Short review: Metabolism of theFusarium mycotoxins deoxynivalenol and zearalenone in plants.
Plants have a high capacity to transform and thereby detoxify deleterious or poisonous compounds, like mycotoxins. The formation of glucose conjugates has a central role in this process. Mammals, however, are able to (partly) release the precursor substances during digestion, reactivating the mycotoxins. This short review provides a brief summary about the metabolism of theFusarium mycotoxins deoxynivalenol and zearalenone in plants. Two examples are discussed in greater detail. First, the formation of deoxynivalenol-3-glucoside in wheat is linked to a quantitative trait locus that is often used forFusarium head blight resistance breeding. Secondly, the metabolism of zearalenone inArabidopsis thaliana results in at least 17 different metabolites, all of which are potentially hazardous for humans and animals.


Prognostic value of the lymph node ratio in breast cancer subtypes.
Several studies indicate that lymph node (LN) ratio (LNR) is more predictive than the traditional LN-positive stratification (pNs). We assessed whether LNR can provide additional prognostic information on node-positive breast cancer patients, particularly with breast cancer subtypes (BCS). We retrospectively reviewed the medical records of 2,049 patients with primary breast cancer treated between January 2006 and December 2011 and identified 511 subjects with positive axillary LN. Clinicopathological findings, types of treatment, and the 5-year overall survival (OS) were included. The results of multivariate analysis of the cohort showed that the correlations of pNs and LNR in regard to OS were P < .001 and P < .001, respectively, with adjustment for tumor characteristics and treatment factors. Moreover, our data revealed that LNR was more predictive in luminal A, luminal B, and luminal human epidermal growth factor receptor 2 in a comparison of pNs and LNR with regard to OS among BCS. LNR and pNs are important prognostic factors with regard to OS for patients with node-positive breast cancer, but LNR has a more correlated value in BCS.


[Long-term results after isolated aortic and mitral valve replacement with a prosthesis (author's transl)].
111 patients with aortic valve replacement (AVR) and 159 patients with mitral valve replacement (MVR) were studied after hospital discharge. The late mortality was 20% after AVR and 32% after MVR. Of the survivors, 93% were greatly improved after AVR and 83% after MVR (functional class I or II). The calculated 5-year survival rate of the patients with AVR is 76% and with MVR 64%. The general state of health of the survivors, the thromboembolic rate and the fatal complications are discussed. Further improvement in the results can only be expected if valve replacement is performed before irreversible myocardial damage has occurred.


Public attitudes toward stuttering in Malaysia.
This study aims to: (a) measure public attitudes toward stuttering in Malaysia using the Public Opinion Survey of Human Attitudes-Stuttering [POSHA-S], (b) determine how reported attitudes and knowledge related to stuttering compare to existing data, and (c) determine whether there are differences between groups for identified variables. A total of 250 adults (mean age = 29 years; range = 19-60 years) completed the POSHA-S in English. We compared this sample's attitudes toward stuttering to POSHA-S data from other global samples. General linear modeling examined differences in overall stuttering score, beliefs, and self reaction subscores for demographic variables such as age, gender, marital status, parenting, education, employment status, prior exposure to a person who stutters, multilingual, race, and religion. The Malaysian participants' overall stuttering score and the beliefs and self reactions subscores were all considerably lower (i.e., less positive) than the other samples around the world from the POSHA-S database median values. Being male, receiving a higher education, and knowing someone who stutters were linked to having more positive self reactions, but none of those factors was linked to positive or negative beliefs. Those who had previously been exposed to stuttering scored significantly higher than those who had not. Malaysians may have less positive attitudes toward stuttering than Westerners. More needs to be done to make society more accepting of people who stutter. Future research should aim to find ways to educate and to raise public awareness about stuttering.


The metzincins--topological and sequential relations between the astacins, adamalysins, serralysins, and matrixins (collagenases) define a superfamily of zinc-peptidases.
The three-dimensional structures of the zinc endopeptidases human neutrophil collagenase, adamalysin II from rattle snake venom, alkaline proteinase from Pseudomonas aeruginosa, and astacin from crayfish are topologically similar, with respect to a five-stranded beta-sheet and three alpha-helices arranged in typical sequential order. The four proteins exhibit the characteristic consensus motif HEXXHXXGXXH, whose three histidine residues are involved in binding of the catalytically essential zinc ion. Moreover, they all share a conserved methionine residue beneath the active site metal as part of a superimposable "Met-turn." This structural relationship is supported by a sequence alignment performed on the basis of topological equivalence showing faint but distinct sequential similarity. The alkaline proteinase is about equally distant (26% sequence identity) to both human neutrophil collagenase and astacin and a little further away from adamalysin II (17% identity). The pairs astacin/adamalysin II, astacin/human neutrophil collagenase, and adamalysin II/human neutrophil collagenase exhibit sequence identities of 16%, 14%, and 13%, respectively. Therefore, the corresponding four distinct families of zinc peptidases, the astacins, the matrix metalloproteinases (matrixins, collagenases), the adamalysins/reprolysins (snake venom proteinases/reproductive tract proteins), and the serralysins (large bacterial proteases from Serratia, Erwinia, and Pseudomonas) appear to have originated by divergent evolution from a common ancestor and form a superfamily of proteolytic enzymes for which the designation "metzincins" has been proposed. There is also a faint but significant structural relationship of the metzincins to the thermolysin-like enzymes, which share the truncated zinc-binding motif HEXXH and, moreover, similar topologies in their N-terminal domains.


Medical inpatients at risk of extended hospital stay and poor discharge health status: detection with COMPRI and INTERMED.
To detect the patients in medical wards at risk of extended LOS and poor discharge health status with the use of complexity prediction instrument (COMPRI) and interdisciplinary medicine (INTERMED) instruments. STUDY 1: In a sample of 275 consecutively admitted medical inpatients, a hierarchical cluster analysis on INTERMED variables was performed. The clusters were compared on length of hospital stay (LOS) and Short Form 36 (SF-36) at discharge. STUDY 2: Receiver operating characteristic (ROC) analysis was used to optimal cut-off points for the COMPRI and INTERMED. Patients detected with COMPRI and INTERMED were then compared with undetected patients on LOS and SF-36. STUDY 1: In concordance with previous findings, a cluster of patients with high biopsychosocial vulnerability was identified with significantly higher scores on LOS (p <.05) and lower scores on SF-36 (p <.001) than patients in other clusters. STUDY 2: A cut-off point for the COMPRI of 5/6 was found to detect patients at risk of long LOS. A cut off score for the INTERMED of 20/21 was found to detect patients at risk of poor discharge health status. Patients detected with COMPRI and INTERMED had a significantly longer LOS (p <.001) and a poorer discharge health status (SF-36 MCS: p <.001; SF-36 PCS: p =.05) than nondetected patients. Of the detected patients, 37% had an extended hospital stay and poor discharge health status; of the nondetected patients, this was only 7%. The COMPRI-INTERMED can help to detect complex patients admitted to medical wards within the first days of admission, and rule out those with a small chance of poor outcomes.


Interleukin 17A exacerbates atherosclerosis by promoting fatty acid-binding protein 4-mediated ER stress in macrophages.
Apoptosis and fatty acid-binding protein-4 (FABP4) induced-endoplasmic reticulum (ER) stress in macrophage is an important pathological process in several vascular occlusive diseases, including atherosclerosis, both of which are accelerated by lipids or inflammatory cytokines. To determine whether interleukin 17A (IL-17A) accelerates atherosclerosis through activating FABP4-mediated ER stress in macrophage. We show here that IL-17A induced ER stress in both murine and human-derived macrophages in vitro, and in the atherosclerotic lesions of ApoE-/- mice. Treating ApoE-/- mice with a chemical chaperone alleviated IL-17A-mediated ER stress and macrophage apoptosis, which was accompanied by recovered atherogenesis. Mechanistically, IL-17A up-regulated the expression of FABP4 (aP2), a cytosolic lipid chaperone that is able to promote lipid-induced macrophage ER stress, through NF-κB and ERK/p38 mitogen-activated protein kinase (MAPK) pathways in macrophages. The inhibition of aP2 expression with a specific chemical inhibitor significantly blocked IL-17A-accelerated ER stress and apoptosis in plaque, and partially rescued IL17A-induced atherogenesis. The data collectively establish a previously unrecognized link between IL-17A and ER stress through cytosolic lipid chaperone aP2 in macrophages and provide a new insight for understanding the role of IL-17A in atherosclerosis.


Vitamin E Ameliorates Impaired Ovarian Development, Oxidative Stress, and Disrupted Lipid Metabolism in Oreochromis niloticus Fed with a Diet Containing Olive Oil Instead of Fish Oil.
Aquaculture feed containing olive oil (OO) instead of fish oil (FO) can cause oxidative stress and impair gonad development in fish. We determined the effect of dietary OO-induced oxidative stress on ovarian development, and explored whether vitamin E (VE) could mitigate negative effects. Female Nile tilapia (Oreochromis niloticus) were fed for 10 weeks with four diets: 5% OO + 70 mg/kg VE, 5% OO + 200 mg/kg VE, 5% FO + 70 mg/kg VE, or 5% FO + 200 mg/kg VE. Dietary OO reduced the specific growth rate and gonadosomatic index, inhibited superoxide dismutase and catalase, delayed ovarian development, decreased serum sex hormone levels, and reduced ovarian triglyceride and n-3 highly unsaturated fatty acid contents. The transcript levels of genes encoding sex hormone receptors (erα, fshr, lhr) and components of the lipid metabolism pathway (pparα, pparγ, hsl, accα, elovl6), the nrf2 signaling pathway (nrf2, keap1), and the nf-κb signaling pathway (nf-κb, tnfα, infγ, il1β) differed between the 70VE/OO and 70VE/FO groups. Supplementation with 200 mg/kg VE mitigated the adverse effects of OO by improving antioxidant capacity and alleviating inflammation and abnormal lipid metabolism. This may be because VE is an antioxidant and it can regulate the nrf2-nf-κb signaling pathway.


A method to determine the protection zone of chemical industrial park considering air quality, health risk and environmental risk: a case study.
In China, chemical enterprises are required to cluster into a large number of chemical industrial parks (CIPs), which increase risks and threats to the environment and human being's health due to aggregation of the complicated chemical process and huge unit scale. Setting a scientific and reasonable protection zone around CIP is a very efficient way to protect surrounding people's health. A method was designed to determine the comprehensive protection zone of CIP, taking into account multiple factors: air quality, health risk and environmental risk. By establishing a comprehensive and multi-levels index system, the protection zone and the corresponding environmental risk management countermeasures can be proposed hierarchically, which are very important to the development and environmental risk management of CIP. A CIP located in coastal area of Shandong Province was studied, and it is turned out that the method to determine the protection zone of chemical industrial park considering air quality, health risk and environmental risk has great advantages compared with other methods.


Mode of protection of mice against herpes simplex virus type 2 infection by Propionibacterium.
We compared various strains of Propionibacterium with regard to protection of young adult mice against lethal infection with herpes simplex virus type 2 (HSV-2). Propionibacterium acnes, P. granulosum, and P. avidum were protective, while P. acidi-propionici and P. lymphophilum were ineffective. The protective effect proved to be in the cell wall fraction. Attempts were made to elucidate possible mechanisms of the protection using both effective and ineffective strains. The results strongly suggest that induction of interferon rather than activation of macrophages and natural killer cells by Propionibacterium pretreatment plays a crucial role, directly or indirectly, in protection against infection by herpes simplex virus. Propionibacterium only moderately protected newborn mice against HSV-2 infection.


Urolithiasis in Okinawa, Japan: a relatively high prevalence of uric acid stones.
The aim of the present study was to investigate the composition of urinary tract stones in patients from Okinawa, the most southern island group of Japan. The study was conducted by 12 hospitals in Okinawa. A total of 1816 urinary tract calculi were obtained from 1816 patients (1323 males; 493 females). The patients had a mean age of 53 +/- 15.3 years (mean +/- SD). The calculi were examined to determine their chemical composition. Stone samples were analyzed by computed infrared spectrophotometer. Pure stones comprised 58.4% of the total, with calcium oxalate stones accounting for 40% (21% monohydrate [whewellite]; 6.6% dihydrate [weddellite]; and 12.4% combined monohydrate and dihydrate stones), uric acid/urate stones for 9.6%, calcium phosphate stones for 5.1%, and struvite stones for 3.7%. The other 41.6% of the stones consisted of calcium oxalate mixed with other components. The male-to-female ratio was 2.7:1. In our series, calcium oxalate stones accounted for 81.6% of the urinary tract calculi, while uric acid/urate stones accounted for 15.8%. Uric acid stones, predominantly the anhydrous and/or dihydrate forms, showed a relatively high prevalence. Calcium oxalate stones, predominantly in the form of whewellite, showed a high prevalence among pure calculi; while the predominant combinations among mixed calculi were weddellite + calcium phosphate and whewellite + uric acid/urate.


Self-expandable metal mesh stents for common bile duct stenosis in chronic pancreatitis: retrospective evaluation of long-term follow-up and clinical outcome pilot study.
Symptomatic common bile duct strictures are frequently seen in the course of severe chronic pancreatitis with a reported incidence of 10-30%. The exact role of endoscopic stenting with metal prostheses as definitive treatment has not yet been clearly defined. Six patients with symptomatic distal common bile duct stricture in the setting of chronic pancreatitis were treated with a self-expandable metal stent in an attempt to achieve permanent drainage. All cases were approached non-operatively and underwent prior unsuccessful treatment with plastic prostheses. The patients received a self-expandable metal Wall stent after a median follow-up of 14 months of plastic stenting. The median follow-up time was 58 months and the stents remained open for a median time of 20 months, respectively. Overall, in 2 of 6 cases the metal stent therapy was successful while 4 of 6 patients required additional endoscopic procedures. Self-expandable metal stents could be useful and adequate in patients who suffer from symptomatic obstructive jaundice due to chronic pancreatitis. However, they provide acceptable dilation of the common bile duct for only a limited period of time. Overall, endoscopic stenting with self-expandable metal stents seems to be an advantageous treatment only for a subgroup of patients who are unfavorable candidates for surgical drainage. To clarify the role of the endoscopic approach, prospective trials with homogeneous and larger patient numbers are necessarily in the future.


Evaluation of engine performance and exhaust emission characteristics in a diesel engine using isobutanol- Calophyllum inophyllum biodiesel-diesel ternary blends.
The availability of natural energy resources and the environmental issues are the most significant issues that are often highlighted by the world communities. With regard to these problems, isobutanol is a higher chain alcohol with four carbons which can be derived from biomass resources and it is potential to become an alternative fuel source besides the biodiesel for a diesel engine. The aim of this study is to evaluate the effect of isobutanol with Calophyllum inophyllum methyl ester and diesel as the ternary blend on physicochemical properties, engine performance, and emission characteristics. Five different fuel blends containing Calophyllum inophyllum biodiesel and isobutanol were tested on a single-cylinder direct injection diesel engine at different engine load of brake mean effective pressure. The physicochemical properties of the fuel blends were measured and then compared with neat diesel. The results indicate that the blend containing isobutanol and CIME gives a slight increase in BSEC and EGT and a minimal drop in BTE as compared to that of neat diesel. Besides that, the tested blends show a reduction of carbon monoxide and unburned hydrocarbon emissions. Meanwhile, all the fuel blends show a minimal increase in carbon dioxide and nitrogen oxides emissions, compared to that of neat diesel. Isobutanol can be proved as a preferred substitute for biodiesel and diesel fuels to achieve desired engine performance and emissions level.


Cellular roadmaps of viroid infection.
Viroids are single-stranded circular noncoding RNAs that infect plants. According to the International Committee on Taxonomy of Viruses, there are 44 viroids known to date. Notably, more than 20 000 distinct viroid-like RNA sequences have recently been identified in existing sequencing datasets, suggesting an unprecedented complexity in biological roles of viroids and viroid-like RNAs. Interestingly, a human pathogen, hepatitis delta virus (HDV), also replicates via a rolling circle mechanism like viroids. Therefore, knowledge of viroid infection is informative for research on HDV and other viroid-like RNAs reported from various organisms. Here, we summarize recent advancements in understanding viroid shuttling among subcellular compartments for completing replication cycles, emphasizing regulatory roles of RNA motifs and structural dynamics in diverse biological processes. We also compare the knowledge of viroid intracellular trafficking with known pathways governing cellular RNA movement in cells. Future investigations on regulatory RNA structures and cognate factors in regulating viroid subcellular trafficking and replication will likely provide new insights into RNA structure-function relationships and facilitate the development of strategies controlling RNA localization and function in cells.


Interleukin-1 receptor antagonist inhibits arthrofibrosis in a post-traumatic knee immobilization model.
Therapies for arthrofibrosis after knee surgery are needed to prevent loss of joint function. Interleukin-1 receptor antagonists (IL-1RA) have shown promise in treating established arthrofibrosis in pilot clinical studies. The objective of this study was to evaluate the ability of intra-articular injection of IL-1RA to prevent knee joint contracture in a post-traumatic knee immobilization model. 20 male Sprague Dawley rats were block randomized into two groups: control and IL-1RA. Rats underwent intra-articular surgical trauma of the right knee with placement of an immobilization suture, securing the knees in 150° flexion. On post-operative days 1 and 8, each group received a 0.1 ml intra-articular injection of either saline (control) or anakinra (IL-1RA:single dosage; 2.63 mg/kg). Rats were euthanized fourteen days after surgery and the immobilization femorotibial angles were measured on the operative limbs with the suture and musculature intact. Subsequently, musculature was removed and femorotibial angles were measured in the operative and non-operative limbs with a defined extension moment applied with the posterior capsule intact or cut. A contracture angle was calculated as the angular difference between the operative and non-operative limb. The immobilization knee flexion angle did not differ (P = 0.761) between groups (control: 152 ± 9; IL-1RA: 150 ± 11). The joint contracture angles (smaller angle = improved outcome) were reduced by 12 degrees on average in the IL-1RA group compared to the control for both the capsule intact (P = 0.024) and cut (P = 0.019) states. Intra-articular IL-1RA injection was found to diminish knee extension deficits associated with arthrofibrosis in a post-traumatic joint immobilization model.


Responses of tropical legumes from the Brazilian Atlantic Rainforest to simulated acid rain.
We investigated the morphological and anatomical effects of simulated acid rain on leaves of two species native to the Brazilian Atlantic Rainforest: Paubrasilia echinata and Libidibia ferrea var. leiostachya. Saplings were subjected to acid rain in a simulation chamber during 10 days for 15 min daily, using H2SO4 solution pH 3.0 and, in the control, deionized water. At the end of the experiment, fragments from young and expanding leaves were anatomically analyzed. Although L. ferrea var. leiostachya leaves are more hydrophobic, rain droplets remained in contact with them for a longer time, as in the hydrophilic P. echinata leaves, droplets coalesce and rapidly run off. Visual symptomatology consisted in interveinal and marginal necrotic dots. Microscopic damage found included epicuticular wax flaking, turgor loss and epidermal cell shape alteration, hypertrophy of parenchymatous cells, and epidermal and mesophyll cell collapse. Formation of a wound tissue was observed in P. echinata, and it isolated the necrosis to the adaxial leaf surface. Acid rain increased thickness of all leaf tissues except spongy parenchyma in young leaves of L. ferrea var. leiostachya, and such thickness was maintained throughout leaf expansion. To our knowledge, this is the first report of acidity causing increase in leaf tissue thickness. This could represent the beginning of cell hypertrophy, which was seen in visually affected leaf regions. Paubrasilia echinata was more sensitive, showing earlier symptoms, but the anatomical damage in L. ferrea var. leiostachya was more severe, probably due to the higher time of contact with acid solution in this species.


Freezing of neural tissues and their transplantation in the brain of rats: technical details and histological observations.
Embryonic neocortical and brainstem tissues were frozen, stored for variable periods, thawed and transplanted into the cerebellum of neonatal host rats. Various conditions related to freezing, media for freezing, DMSO as the cryoprotectant, and thawing were analyzed. The findings indicated that the following conditions yielded best results for neocortical transplantation: freezing at a rate of 1 degrees C/min, using rat amniotic fluid as the medium for freezing, using 10% DMSO as the cryoprotectant, storing the frozen tissues at -90 degrees C, thawing the tissues fast just prior to transplantation, and transplanting them in the host brain with little or no delay. Other conditions having adverse effects on the neural tissues were considered. Issues pertaining to transplantability and retainability of the neural tissues inside the host brain, and effects of freezing and thawing on the long-term viability of the neural tissues and their growth are discussed.


A 10-year, single-institution analysis of clinicopathologic features and sentinel lymph node biopsy in thin melanomas.
The 2009 American Joint Committee on Cancer criteria for thin cutaneous melanomas recommend staging sentinel lymph node (SLN) biopsy (SLNB) for select stage IB tumors. SLNB in this population remains controversial because of low rates of node positivity and inconsistent prognostic parameters. The purpose of this study was to examine the association between multiple clinicopathologic features and SLNB result, and clinical outcome. Clinical and pathologic parameters from patients with melanomas less than or equal to 1.00 mm receiving wide local excision with SLNB at our institution from 2001 through 2010 were recorded. Analysis for any statistically significant relationships between recorded parameters and SLN results and outcome were performed. A total of 189 cases yielded 3 positive SLNBs (1.6%). Disease progression occurred in 6 cases (3.2%). Positive SLNB predicted distant metastasis and death from disease (P = .0017). Mitotic rate was not associated with a positive SLNB result. The follow-up time for this study was limited (mean = 40.7 months). Our data confirm a statistically significant relationship between SLNB result and likelihood for distant metastasis in thin melanoma. There was a trend for a relationship between mitotic rate and clinical outcome. This relationship reached statistical significance at a mitotic rate of greater than 3 mitoses/mm(2).


Low rectovaginal fistulas. Approach and treatment.
Experience with thirty-two patients with a low rectovaginal fistula with or without attendant sphincter damage is reported. The technic used is advancement of the anterior rectal wall with excision of the infected anal glandular tissue and repair of muscle tissue when indicated. Anorectal infection and childbirth injuries were the common causes. An acceptable recurrence rate was achieved. Colostomy was not used in this series.


Survival rate and failure characteristics for two post designs.
The dental records of 638 patients treated with 788 posts and cores were studied to analyze failure rate and failure characteristics for two post designs. Frequency of the technical failures, loss of retention, root fracture, and post fracture were recorded 4 to 5 years after post cementation. There were two types of posts: custom-cast tapered posts and parallel-sided serrated posts. The cumulative failure rate was 15% for 456 tapered posts and 8% for 332 Para-Post posts. Loss of retention was the most frequent reason for failure for both types of posts, whereas root fractures had the most serious consequences, and all resulted in extraction. A significantly higher success rate was recorded for parallel-sided serrated posts, compared with custom-cast posts, regarding the total failure rate and the severity of the failure (p < 0.05).


The relationship between coherence and nonlinear characteristics in Renshaw cell responses to random motor axon stimulation.
Cat spinal Renshaw cells were activated by stimulating muscle nerves or ventral roots with random (pseudo-Poisson) patterns of brief electrical stimuli. This input pattern is optimal for a comparative study in both the frequency- and time-domain. The frequency-dependent variable of particular interest in this study was the coherence as a measure of the degree to which signal transmission is linear and noise-free; it was estimated via spectral analysis. Time-domain analysis consisted of calculating peri-stimulus time histograms in order to estimate the amount of nonlinearity in the cell responses to pairs of stimuli. The main result was that the amount of nonlinearity measured in this way did not profoundly depress the coherence. Two types of peri-stimulus time histogram were calculated: the "conventional" peri-stimulus time histogram (as a reference) computed with respect to all the stimuli in a train, and the "conditional" peri-stimulus time histogram computed with respect to the second in pairs of stimuli which were separated from each other by varied intervals delta. The latter type of peri-stimulus time histogram showed that Renshaw cell responses to stimuli were conditioned by preceding stimuli, which could facilitate (at small delta s) and/or more often depress (up to several hundreds of milliseconds) the subsequent responses in a nonlinear manner. The objective of this study was to test the hypothesis that nonlinear characteristics contribute significantly to depress the coherence from its optimal value (1).(ABSTRACT TRUNCATED AT 250 WORDS)


Metabolic Reprogramming of Immune Cells in Cancer Progression.
Immune cells play a key role in host defense against infection and cancer. Upon encountering danger signals, these cells undergo activation leading to a modulation in their immune functions. However, recent studies reveal that immune cells upon activation also show distinct metabolic changes that impact their immune functions. Such metabolic reprogramming and its functional effects are well known for cancer cells. Given that immune cells have emerged as crucial players in cancer progression, it is important to understand whether immune cells also undergo metabolic reprogramming in tumors and how this might affect their contribution in cancer progression. This emerging aspect of tumor-associated immune cells is reviewed here, discussing metabolic reprogramming of different immune cell types, the key pathways involved, and its impact on tumor progression.


Transient receptor potential channel 3 in human liver and gallbladder - An investigation in body donors.
Since the discovery of TRP proteins in 1969, during studies of the fruit fly Drosophila melanogaster, interest around them and the subfamily of TRPC channels has remained high. TRPC3 was able to be detected in a number of organs in rodents, such as rats and mice, and also in various human tissues. For the most part, these investigations were carried out using gene expression of TRPC3. Further work has already confirmed the relevance of TRPC3 in the context of neurodegenerative diseases, such as spinocerebellar ataxia, and carcinogenic entities, such as ovarian carcinoma. An association with TRPC3 has also been demonstrated for diseases that affect the liver. In order to confirm the expression of TRPC3 in the human liver, this study uses samples taken from eight (n = 8) fixated human body donors and analyzed with immunohistochemistry. In accordance with the macroscopic anatomy of the organs, six samples (n = 6) of liver tissue and three (n = 3) of gallbladder tissue were obtained. TRPC3 was clearly detected in all liver and gallbladder samples examined. Thus, it is not unlikely that TRPC3 plays a role in the extensive metabolic processes of the liver and could also serve as a target for pharmacological interventions in an imbalance of calcium homeostasis.


Leptospirosis outbreak in Tanzania: An alarming situation.
On July 5, 2022, the Tanzanian Ministry of Health (MoH) announced the re-emergence of leptospirosis after reporting 20 confirmed symptomatic cases and 3 mortalities. Leptospirosis is caused by a spirochete bacterium that lives in an animal's renal tubule and spreads to individuals through contact with contaminated animal urine. Unsupervised agricultural practices, urban development, wildlife infiltration, and a lack of sanitation have all been proposed as potential environmental causes of the present outbreak. The MoH is taking the necessary steps to halt the spread of said outbreak with assistance from the World Health Organization (WHO). This article examines the risk factors, etiology, number of confirmed cases, and subsequent case index to analyse the epidemiology of the current leptospirosis outbreak in Tanzania's southern Linda region. In light of these findings, this research further details recent recommendations made by the WHO, Centers for Disease Control and Prevention, and MoH to mitigate such an alarming situation. These recommendations include early detection and isolation, contact tracing, and chemoprophylaxis using doxycycline. The article concludes by outlining suggestions for individuals and governments, including the launch of public awareness campaigns, immunisation, increased surveillance, rapid detection testing, and the installation of suitable purification systems, to help contain future leptospirosis outbreaks.


Algorithm-mediated social learning in online social networks.
Human social learning is increasingly occurring on online social platforms, such as Twitter, Facebook, and TikTok. On these platforms, algorithms exploit existing social-learning biases (i.e., towards prestigious, ingroup, moral, and emotional information, or 'PRIME' information) to sustain users' attention and maximize engagement. Here, we synthesize emerging insights into 'algorithm-mediated social learning' and propose a framework that examines its consequences in terms of functional misalignment. We suggest that, when social-learning biases are exploited by algorithms, PRIME information becomes amplified via human-algorithm interactions in the digital social environment in ways that cause social misperceptions and conflict, and spread misinformation. We discuss solutions for reducing functional misalignment, including algorithms promoting bounded diversification and increasing transparency of algorithmic amplification.


Forest 404: Using a BBC drama series to explore the impact of nature's changing soundscapes on human wellbeing and behavior.
Extensive ecosystem degradation and increasing urbanization are altering human relationships with nature. To explore these trends, we created a transdisciplinary, narrative-led podcast series produced by the BBC, called Forest 404. The series explored the implications of a world without nature. An online experimental component mobilized audience participation (n = 7,596) to assess responses to natural soundscapes with and without abiotic, biotic, and poetic elements across five biomes. Conditions featuring the sounds of wildlife, such as bird song, were perceived to be more psychologically restorative than those without. Participants' personal lived experiences were strongly related to these outcomes; those who had memories triggered by the sounds were more likely to find them psychologically restorative and exhibited a greater motivation to preserve them. Moreover, the effects of both soundscape composition and memories on preservation behavior were partially mediated by restorative potential; respondents were more likely to want to protect the sounds they heard if they thought they might offer therapeutic outcomes. Our findings highlight the value of art-science collaborations and demonstrate how maintaining contact with the natural world can promote wellbeing and foster behaviors that protect planetary health.


Protein chemical shift assignments of the unbound and RNA-bound forms of the alternative splicing factor SUP-12 from C. elegans.
The splicing factor SUP-12 from Caenorhabditis elegans binds to regulatory RNA elements in pre-mRNA in order to generate tissue-specific alternative splicing for genes such as the fibroblast growth factor receptor egl-15. In nematode muscle cells, SUP-12 promotes the use of a mutually exclusive exon to impart variant binding specificity to the EGL-15 extracellular protein domain. Here we report the side chain and backbone (1)H, (13)C and (15)N chemical shift assignments for the bacterially expressed RNA recognition motif domain from SUP-12, both in isolation as well as bound to a short RNA derived from the intron sequence between exon 4 and exon 5B of egl-15. Comparison of protein chemical shift values for both the backbone and side chain nuclei, coupled with secondary chemical shift analysis, reveal initial details of the RNA recognition.


Quantal analysis estimates docking site occupancy determining short-term depression at hippocampal glutamatergic synapses.
Before fusion, synaptic vesicles (SVs) pause at discrete release/docking sites. During repetitive stimulation, the probability of site occupancy changes following SV fusion and replenishment. The occupancy probability is considered to be one of the crucial determinants of synaptic strength, but it is difficult to estimate separately because it usually blends with other synaptic parameters. Thus, the contribution of site occupancy to synaptic function, particularly to synaptic depression, remains elusive. Here, we directly estimated the occupancy probability at the hippocampal mossy fibre-CA3 interneuron synapse showing synaptic depression, using statistics of counts of vesicular events detected by deconvolution. We found that this synapse had a particularly high occupancy (∼0.85) with a high release probability of a docked SV (∼0.8) under 3 mm external calcium conditions. Analyses of quantal amplitudes and SV counts indicated that quantal size reduction decreased the amplitudes of all responses in a train to a similar degree, whereas release/docking site number was unchanged during trains, suggesting that quantal size and release/docking site number had little influence on the extent of synaptic depression. Model simulations revealed that the initial occupancy with high release probability and slow replenishment determined the time course of synaptic depression. Consistently, decreasing external calcium concentration reduced both the occupancy and release probability, and the reductions in turn produced less depression. Based on these results, we suggest that the occupancy probability is a crucial determinant of short-term synaptic depression at glutamatergic synapses in the hippocampus. KEY POINTS: The occupancy probability of a release/docking site by a synaptic vesicle at presynaptic terminals is considered to be one of the crucial determinants of synaptic strength, but it is difficult to estimate separately from other synaptic parameters. Here, we directly estimate the occupancy probability at the hippocampal mossy fibre-interneuron synapse using statistics of vesicular events detected by deconvolution. We show that the synapses have particularly high occupancy (0.85) with high release probability (0.8) under high external calcium concentration ([Ca2+ ]o ) conditions, and that both parameter values change with [Ca2+ ]o , shaping synaptic depression. Analyses of the quantal amplitudes and synaptic vesicle counts suggest that quantal sizes and release/docking site number have little influence on the extent of synaptic depression. The results suggest that the occupancy probability is a crucial determinant of short-term synaptic depression at glutamatergic synapses in the hippocampus.