Nanotoxicity assessment in plants: an updated overview.
Nanotechnology is rapidly emerging and innovative interdisciplinary field of science. The application of nanomaterials in agricultural biotechnology has been exponentially increased over the years that could be attributed to their uniqueness, versatility, and flexibility. The overuse of nanomaterials makes it crucial to determine their fate and distribution in the in vitro (in cell and tissue cultures) and in vivo (in living species) biological environments by investigating the nano-biointerface. The literature states that the beneficial effects of nanoparticles come along with their adverse effects, subsequently leading to an array of short-term and long-term toxicities. It has been evident that the interplay of nanoparticles with abiotic and biotic communities produces several eco-toxicological effects, and the physiology and biochemistry of crops are greatly influenced by the metabolic alterations taking place at cellular, sub-cellular, and molecular levels. Numerous risk factors affect nanoparticle's accumulation, translocation, and associated cytogenotoxicity. This review article summarizes the contributing factors, possible mechanisms, and risk assessment of hazardous effects of various types of nanoparticles to plant health. The methods for evaluating the plant nanotoxicity parameters have been elaborated. Conclusively, few recommendations are put forward for designing safer, high-quality nanomaterials to protect and maintain environmental safety for smarter agriculture demanded by researchers and industrialists.


Oxidative stress in patients with chronic heart failure and type 2 diabetes mellitus.
Parameters of oxidative stress were studied in patients with chronic heart failure and/or type 2 diabetes mellitus. Chronic heart failure was accompanied by severe oxidative stress, while in patients with type 2 diabetes mellitus the signs of oxidative stress were less pronounced. The intensity of free radical oxidation in patients with chronic heart failure and type 2 diabetes mellitus was not higher compared to patients with chronic heart failure.


Questions elderly patients have about on-going therapy: a pilot study to assist in communication with physicians.
This pilot study examined the prevalence and types of questions elderly patients have about their current drug therapy. It also evaluated the effectiveness of a brief intervention to prepare patients to ask questions about drug therapy during medical visits. The research used a posttest-only experimental design. Forty-five elderly patients seen at a primary care clinic during a one-month period consented to participate and completed the study. After consent, subjects were randomly assigned to intervention and control conditions. A brief interview with intervention group subjects conducted by a medical student assigned to the clinic as part of a summer research experience helped subjects formulate questions they had about current therapy before they went into medical visits. Patient-physician visits were audiotaped and patient questions about medications and health care were tallied. Subjects in the intervention group were significantly more likely to ask questions of providers than were subjects in the control group. Intervention group subjects were found to ask a wider variety of medication-related questions than were control group subjects, including questions related to proper use, problems perceived with medications, and effectiveness of treatment. Assisting patients to formulate questions before medical visits results in an increased likelihood that patients will ask questions and will ask a wider variety of questions during the medical visit.


Surfactant-induced release of liposomal contents. A survey of methods and results.
A systematic approach to the phenomenon of surfactant-dependent release of liposomal contents has been attempted. A variety of methods have been comparatively studied. The influence of the size of the entrapped molecule, nature of the surfactant, composition of bilayers and sonication of liposomes have been considered separately. In order to compare different results, a parameter has been defined, R50, as the phospholipid/surfactant mole ratio producing 50% release of the entrapped solute. This parameter appears to be, to a large extent, independent of time and liposome concentration. Surfactant-induced release of liposomal contents does not occur as a result of breakdown of phospholipid bilayers, but is rather a different phenomenon, occurring at detergent concentrations substantially lower (2-5 times) than solubilization. The required amount of surfactant appears to increase with the size of the entrapped solute. R50 depends clearly on the nature of the soluble amphiphile, but there is no obvious relationship with its critical micellar concentration. Liberation of vesicle content also depends on bilayer composition: phospholipids have various effects on the stability of the membrane, while the hydrophobic peptide, gramicidin A, appears to have little influence. Cholesterol is interesting, since at equimolar proportions with phosphatidylcholine, it decreases the stability of bilayer towards Triton X-100, while increasing it in the presence of cholate. Sonication also exerts an influence on the surfactant-dependent release of vesicle contents; it appears to decrease the bilayer stability, so that lower detergent concentrations are required to liberate the entrapped solutes. Finally, it should be noted that, although the decrease in self-quenching of 6-carboxyfluorescein is a convenient method for the study of solute liberation, glucose release, as detected by enzymatic methods, may be more reliable for accurate measurements.


[Hydrocarbon pneumonitis. A case report].
Hydrocarbon pneumonitis has become rare in industrialized countries. However, it remains one of the most frequent emergencies in African dispensaries, especially within children, by accidental ingestion. The case reported here is about a diesel aspiration after siphoning. As in most cases, the healing occurred without any complication.


Toxoplasma gondii binds sheep prolactin.
Taking into account the literature reports on the involvement of prolactin (PRL) in the regulation of immunity against Toxoplasma gondii, we decided to check whether this parasite has the ability to bind the lactotrophic hormone. We examined T. gondii binding of sheep fluoresceine- and biotine-labeled prolactin isolated from pituitary (shPRL). In this work we announced for the first time that shPRL was bound to live tachyzoites of RH (type I) and ME49 (type II) strains. Furthermore, by use of competitive inhibition analysis, we confirmed that this binding was specific for both tested T. gondii strains.


Diversity of endosymbiotic Nostoc in Gunnera magellanica from Tierra del Fuego, Chile [corrected].
Global warming is causing ice retreat in glaciers worldwide, most visibly over the last few decades in some areas of the planet. One of the most affected areas is the region of Tierra del Fuego (southern South America). Vascular plant recolonisation of recently deglaciated areas in this region is initiated by Gunnera magellanica, which forms symbiotic associations with the cyanobacterial genus Nostoc, a trait that likely confers advantages in this colonisation process. This symbiotic association in the genus Gunnera is notable as it represents the only known symbiotic relationship between angiosperms and cyanobacteria. The aim of this work was to study the genetic diversity of the Nostoc symbionts in Gunnera at three different, nested scale levels: specimen, population and region. Three different genomic regions were examined in the study: a fragment of the small subunit ribosomal RNA gene (16S), the RuBisCO large subunit gene coupled with its promoter sequence and a chaperon-like protein (rbcLX) and the ribosomal internal transcribed spacer (ITS) region. The identity of Nostoc as the symbiont was confirmed in all the infected rhizome tissue analysed. Strains isolated in the present study were closely related to strains known to form symbioses with other organisms, such as lichen-forming fungi or bryophytes. We found 12 unique haplotypes in the 16S rRNA (small subunit) region analysis, 19 unique haplotypes in the ITS region analysis and 57 in the RuBisCO proteins region (rbcLX). No genetic variability was found among Nostoc symbionts within a single host plant while Nostoc populations among different host plants within a given sampling site revealed major differences. Noteworthy, interpopulation variation was also shown between recently deglaciated soils and more ancient ones, between eastern and western sites and between northern and southern slopes of Cordillera Darwin. The cell structure of the symbiotic relationship was observed with low-temperature scanning electron microscopy, showing changes in morphology of both cyanobiont cells (differentiate more heterocysts) and plant cells (increased size). Developmental stages of the symbiosis, including cell walls and membranes and EPS matrix states, were also observed.


Pneumocystis carinii pneumonia following heart transplantation.
Pneumocystis carinii pneumonia represents a rare complication that is associated with a high mortality following heart transplantation. The cases of two heart transplant recipients who developed Pneumocystis pneumonia within the first 3 postoperative months are reported. Both patients had severe clinical symptoms of the disease; the diagnosis was confirmed by bronchoalveolar lavage, and the patients were treated with a combination of trimethoprim and sulfamethoxazole. Both patients recovered and are well at the time of this report.


Circulating leptin and body composition in chronic obstructive pulmonary disease.
Nutritional depletion and weight loss are two features of chronic obstructive pulmonary disease (COPD), and the association between low body mass index (BMI) and poor prognosis in patients with COPD is a common clinical observation. Mechanisms of weight loss are still unclear in COPD. Excessive energy expenditure partly due to increased work of breathing was shown, but other mechanisms have been searched for. Leptin is a hormone secreted by adipocytes that plays an important role in energy homeostasis and regulates body weight through control of appetite and energy expenditure. The aim of this study was to evaluate the association of circulating leptin levels and measures of body composition in COPD patients. Thirty male COPD outpatients (mean age 66.3 +/- 8.4) and 20 controls (mean age 65.9 +/- 10.8) were included in the study. After standard spirometry and body composition measurements, serum leptin concentration was measured by ELISA assay. COPD patients were grouped according to BMI. Mean BMI was 19.01 +/- 2.26 kg/m2 in group 1 (COPD patients with low BMI), 26.85 +/- 4.51 in group 2 COPD (COPD patients with normal/high BMI) and 27.64 +/- 2.75 kg/m2 in healthy controls (group 3). Mean serum leptin concentration was 1.41 +/- 1.86 ng/ml in group 1, 2.60 +/- 1.38 ng/ml in group 2 and 2.82 +/- 1.46 ng/ml in group 3 (p = 0.002). Leptin correlated to not only BMI but also body weight, waist circumference, triceps and biceps skinfold thickness and body fat percent (p < 0.05 for all). Results of this study suggest that the cause of weight loss is not increased circulating leptin in COPD. Instead, leptin remains regulated in COPD and further decreased in patients with low BMI, probably as a compensatory mechanism to preserve body fat content, which should be evaluated in further studies.


Aneurysmal subarachnoid haemorrhage in Māori and European New Zealanders - A comparative study.
A retrospective analysis of 358 patients admitted to the neurosurgical unit at a tertiary referral centre in Aotearoa New Zealand between 2010 and 2017 was undertaken to describe the relationship of ethnicity to demographic, disease characteristic and outcome data in Māori and European New Zealanders experiencing aneurysmal subarachnoid haemorrhage (aSAH). Māori had a higher incidence (RR 1.38, p = 0.01; 95 % CI 1.08-1.77) and presented at a younger age (mean age difference of 5 years). Higher rates of smoking and hypertension were observed in the Māori population studied. However, these do not fully explain any other differences observed. There was no significant difference in clinical grade at presentation. However, despite experiencing lower rates of posterior circulation aneurysms (adjusted OR 0.33, p = 0.05) and radiological findings predicting highest risk of vasospasm i.e., Modified Rankin Scale 4 (OR 0.54, p = 0.02), Māori had a higher incidence of clinical vasospasm (adjusted OR 1.40, p = 0.048, 95 % CI 1.01-1.98). While there was no observed difference in mortality, Māori may experience lower rates of excellent neurological recovery and survive with greater disability compared to European New Zealanders. Overall, this study is suggestive of inequities between Māori and European New Zealanders and highlights the need for further investigation in this area.


Synthesis of fused 4,5-disubstituted indole ring systems by intramolecular friedel-crafts acylation of 4-substituted indoles.
4-Substituted indoles containing a variety of electrophiles and N-substituents undergo Friedel-Crafts acylation to give exclusively the products of cyclization at the 5-position of indole. These indanones and tetralones have been scarcely prepared and are subunits in natural products and analogues of potential biological significance.


Single-cell RNA sequencing reveals small extracellular vesicles derived from malignant cells that contribute to angiogenesis in human breast cancers.
Breast cancer is the most common cancer affecting women across the world. Tumor endothelial cells (TECs) and malignant cells are the major constituents of the tumor microenvironment (TME), but their origin and role in shaping disease initiation, progression, and treatment responses remain unclear due to significant heterogeneity. Tissue samples were collected from eight patients presenting with breast cancer. Single-cell RNA sequencing (scRNA-seq) analysis was employed to investigate the presence of distinct cell subsets in the tumor microenvironment. InferCNV was used to identify cancer cells. Pseudotime trajectory analysis revealed the dynamic process of breast cancer angiogenesis. We validated the function of small extracellular vesicles (sEVs)-derived protein phosphatase 1 regulatory inhibitor subunit 1B (PPP1R1B) in vitro experiments. We performed single-cell transcriptomics analysis of the factors associated with breast cancer angiogenesis and identified twelve subclusters of endothelial cells involved in the tumor microenvironment. We also identified the role of TECs in tumor angiogenesis and confirmed their participation in different stages of angiogenesis, including communication with other cell types via sEVs. Overall, the research uncovered the TECs heterogeneity and the expression levels of genes at different stages of tumor angiogenesis. This study showed sEVs derived from breast cancer malignant cells promote blood vessel formation by activating endothelial cells through the transfer of PPP1R1B. This provides a new direction for the development of anti-angiogenic therapies for human breast cancer.


An association study of clock genes with major depressive disorder.
To study the relationship between clock genes and Major Depressive Disorder (MDD). GEO database was used to obtain the chip data and clinical information of datasets GSE98793, GSE39653 and GSE52790. The differentially expressed clock genes were found through the analysis of the differentially expressed genes between MDD and healthy controls. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes Pathway (KEGG) enrichment analysis were performed on the differential expressed clock genes. Lasso Regression and Support Vector Machine (SVM) method were used for screening the differential expressed clock genes. Logistic regression was used to establish a diagnostic model for depression with the screened genes. Receiver Operating Characteristic (ROC) Curve was used to verify the model. Gene differential expression analysis was performed for MDD with high scores and MDD with low scores in the diagnostic model. Gene Set Enrichment Analysis (GSEA) enrichment analysis was performed for differentially expressed genes. Single-gene GSEA was used to analyze each gene in the model separately. Cibersort method was used to analyze the immune infiltration of MDD and healthy controls, and the correlation between immune cells and clock genes was analyzed. Cytoscape was used to analyze the clock gene interaction network. The DGIdb website was used to predict potentially effective therapeutic drugs for clock genes closely related to MDD. Six genes were identified by differential expression analysis of clock genes between MDD and healthy controls. GO and KEGG enrichment analysis of 6 genes showed that their pathways were concentrated such as circadian rhythm, rhythmic process, TGF - beta signaling pathway, longevity regulating pathway-multiple species, adipocytokine signaling pathway and so on. Lasso regression and SVM were used to screen out 5 clock genes (HDAC1, ID3, NFIL3, PRKAA1, TNF) for MDD. The diagnostic model of depression was established according to the 5 clock genes. The area under the curve (AUC) of the established depression diagnostic model was 0.686. Gene difference analysis was performed between MDD patients with high score of clock gene diagnostic model and MDD patients with low score. GSEA was performed for the differential genes showed that the most enriched pathways were:adipocytokine signaling pathway, TGF beta signaling pathway, oxidative phosphorylation, primary immunodeficiency, and so on. The single gene GSEA showed that the most enriched pathways were Toll like receptor signaling pathway, glucolipid metabolism, amino acid metabolism, neuroactive ligand receptor interaction, and so on. The results of immune infiltration analysis showed that NK cells resting and Macrophages M2 were different between MDD and control groups. In MDD, the gene closely related to NK cells resting was HDAC1, and the genes closely related to Macrophages M2 were HDAC1 and NFIL3. The RNA interactions network of clock genes shows that the regulation process is complex, which can provide a reference for subsequent related research. Potential therapeutic drugs predict display, among the 5 clock genes, TNF, HDAC1, and PRKAA1 may have potential effective therapeutic drugs. Among all CLOCK genes, HDAC1, ID3, NFIL3, PRKAA1, TNF are closely related to MDD. Among them, TNF, HDAC1, and PRKAA1 may have potential effective therapeutic drugs.


Biomechanical analysis for stress fractures of the anterior middle third of the tibia in athletes: nonlinear analysis using a three-dimensional finite element method.
We evaluated stresses in the anterior middle third of the tibia that have been reported to predict a poor prognosis for tibial stress fractures compared to other predominant sites (posteromedial regions of the distal third and proximal third). The effect of two different loads (bending-compression load and torsional load) on three sites was investigated using a three-dimensional finite element method. The model was constructed using the tibia, fibula, proximal tibiofibular joint, interosseous membrane, and tibiofibular ligament based on computed tomography scans obtained at 4-mm intervals of the lower leg of a 20-year-old woman who exhibited no abnormal findings on roentgenograms. First, a normal model was constructed using normal material properties, and then the model was modified to produce fracture models by varying the mechanical properties of each predominant site and expanding the area in three gradual phases on the assumption that the fracture advanced in three phases. Each model was tested against the same two loads, and stresses at the nodal points on the border of the fracture area and normal area were compared in each cross section to determine the effect of the load on fracture advancement. In response to torsional load, both the normal model and fracture models tended to show higher values for the posteromedial distal third than the anterior middle third. By examining the bending-compression load it could be seen that the mean peak value significantly decreased between the first and second phases in fracture models of the anterior middle third. This finding was inconsistent with our previous belief that the bending-compression load would have more serious consequences than the torsional load. In contrast, when the area of fracture was expanded into the third phase, maximum values were significantly higher than during the second phase. No similar finding was observed for the posteromedial distal third, suggesting that the anterior middle third may have the same stable biomechanical conditions as the posteromedial distal third at an earlier stage and thus have little influence on fractures. When the fracture is more advanced, however, the conditions change suddenly, and a bending-compression load may adversely affect the mechanical conditions in this area and thereby cause complete fracture.


Clinical diagnosis and genetic counseling of atypical ataxia‑telangiectasia in a Chinese family.
Ataxia‑telangiectasia (A‑T) is an autosomal recessive chromosome breakage disorder caused by mutations in the ATM serine/threonine kinase (ATM) gene. Typically, it presents in early childhood with progressive cerebellar dysfunction, accompanied by immunodeficiency and oculocutaneous telangiectasia. In the present study, the clinical and genetic findings of a Chinese family affected with A‑T in two live siblings, the proband (II‑2) and his elder brother (II‑1), as well as a fetus (II‑3) were reported. General health, clinical neurological, electrophysiological (motor and sensory nerve conduction) and magnetic resonance imaging evaluations revealed that patients II‑1 and II‑2 had similar symptoms of ataxia, dysarthria, conjunctival hyperemia and elevated serum α‑fetoprotein, whereas patient II‑1 had earlier A‑T onset at 2 years old and more serious problems with movement and intelligence. Targeted sequencing followed by Sanger sequencing revealed that these two patients carried the compound heterozygotes of a novel nonsense mutation c.5170G>T (p.Glu1724Ter) and a known nonsense mutation c.748C>T (p.Arg250Ter) in the ATM gene. Each mutation was inherited from an asymptomatic parent, which therefore confirmed the diagnosis of A‑T. Given this, proband's mother performed prenatal diagnosis in her third pregnancy. Unfortunately, the fetus had the same causal mutations as its siblings and the pregnancy was terminated. The findings of the present study expanded the mutation spectrum of the ATM gene and may help in understanding the genetic basis of A‑T, in order to guide genetic counseling and prenatal diagnosis.


Diversity and biological characteristics of macrofungi of district Bajaur, a remote area of Pakistan in the Hindu Kush range.
The present study was carried out to document the diversity and ecological characteristics of macrofungi of Bajaur, Pakistan. The diversity of macrofungi comprised 51 species belonging to 22 families and 37 genera. The families Agaricaceae (7 species) and Psathyrellaceae (7 species) were found dominant followed by Tricholomataceae (4 species), Fomitopsidaceae and Polyporaceae (4 species each) and Amanitaceae (3 species). White (23 species), brown (11 species), and yellow were the most prevalent morphological colours in basidiocarps (8 species). Among the identified species, 32 were saprophytic in nutrition followed by 7 parasitic, 6 saprophytic and parasitic both, while 6 mycorrhizal that make association with higher plants. The distribution of macrofungal species in the three tehsils of Bajaur was also evaluated based on Shannon diversity index, Simpson diversity index and evenness. The highest Shannon diversity index and Simpson diversity index were found for tehsil Utman Kheil at 3.73 and 0.97, while the maximum value of evenness for tehsil Khar with 0.92 value. The results indicate a very high species richness of the study site. Four species out of the total were identified to be new reports from Pakistan. This survey's findings suggested that there is a wide variety of macrofungi that might be used as food and alternative medications if further research is carried out.


Evidence-based mental health policy: a critical appraisal.
Arguments for and against evidence-based psychiatry have mostly centred on its value for clinical practice and teaching. Now, however, use of the same paradigm in evaluating health care has generated new problems. To outline the development of evidence-based health care; to summarise the main critiques of this approach; to review the evidence now being employed to evaluate mental health care; and to consider how the evidence base might be improved. The following sources were monitored: publications on evidence-based psychiatry and health care since 1990; reports of randomised trials and meta-analytic reviews to the end of 2002; and official British publications on mental health policy. Although evidence-based health care is now being promulgated as a rational basis for mental health planning in Britain, its contributions to service evaluation have been distinctly modest. Only 10% of clinical trials and meta-analyses have been focused on effectiveness of services, and many reviews proved inconclusive. The current evidence-based approach is overly reliant on meta-analytic reviews, and is more applicable to specific treatments than to the care agencies that control their delivery. A much broader evidence base is called for, extending to studies in primary health care and the evaluation of preventive techniques.


Intelligence as a predictor of nonverbal learning with learning-disabled children.
Although the notion that the WISC-R measures children's learning proficiency is widespread, there are few data to support this hypothesis. The present study concerned the prediction of learning proficiency on the basis of scores on the WISC-R with 60 children referred because of classroom learning difficulties. The single best predictor, WISC-R Coding, accounted for over 50% of the variance in a controlled nonverbal paired associate learning measure. The multiple correlations for the dependent measures of trials to criteria and number of correct responses were in the 90s. In each case, the Coding and Similarities subtests, with Vocabulary in opposition, proved to be the most potent predictors of nonverbal learning.


Inactivation of calcium channels in vascular smooth muscle myocytes.
Many of the structural domains involved in Ca2+ channel (CACN) inactivation are also involved in determining their sensitivity to antagonist inhibition. We hypothesize that differences in inactivation properties and their structural determinants may suggest candidate domains as targets for the development of novel, selective antagonists. The characteristics of Ca2+ current (ICa) inactivation, steady-state inactivation (SSIN), and recovery from inactivation were studied in freshly dispersed smooth muscle cells from rabbit portal vein (RPV) using whole-cell, voltage-clamp methods. The time course of inactivation could be represented by two time constants. Increasing ICa by increasing [Ca2+]o or with more negative holding potentials decreased both time constants. With Sr2+, Ba2+, or Na+ as the charge carrier, ICa inactivation was also represented by two time constants, both of which were larger than those found with Ca2+. With Ca2+, Sr2+, or Ba2+ as the charge carrier, both time constants had minimum values near the voltage associated with maximum current. When Na+ (140 mM) was the charge carrier, voltages for Imax (-20 mV) or taumin (0 mV) did not correspond. SSIN of ICa had a half-maximum voltage of -32 +/- 4 mV for Ca2+, -43 mV +/- 5 mV for Sr2+, -41 +/- 5 mV for Ba2+, and -68 +/- 6 mV for Na+. The slope factor for SSIN per e-fold voltage change was 6.5 +/- 0.2 mV for Ca2+, 6.8 +/- 0.3 for Sr2+, and 6.6 +/- 0.2 for Ba2+, representing four equivalent charges. When Na+ or Li+ was the charge carrier, the slope factor was 13.5 +/- 0.7 mV, representing two equivalent charges. For ICa in rat left ventricular (rLV) myocytes, there was no difference in the slope factor of SSIN for Ca2+ and Na+. The rate of recovery of ICa from inactivation varied inversely with recovery voltage and was independent of the charge carrier. These results suggest that inactivation of ICa in PV myocytes possess an intrinsic voltage dependence that is modified by Ca2+. For RPV but not rLV ICa, the charge of the permeating ion confers the voltage-dependency of SSIN.


Smart and automation technologies for ensuring the long-term operation of a factory amid the COVID-19 pandemic: an evolving fuzzy assessment approach.
The COVID-19 pandemic has severely impacted factories all over the world, which have been closed to avoid the spread of COVID-19. As a result, ensuring the long-term operation of a factory amid the COVID-19 pandemic becomes a critical but challenging task. To fulfill this task, the applications of smart and automation technologies have been regarded as an effective means. However, such applications are time-consuming and budget-intensive with varying effects and are not necessarily acceptable to workers. In order to make full use of limited resources and time, it is necessary to establish a systematic procedure for comparing various applications of smart and automation technologies. For this reason, an evolving fuzzy assessment approach is proposed. A case study has been conducted to demonstrate the effectiveness of the evolving fuzzy assessment approach in ensuring the long-term operation of a factory amid the COVID-19 pandemic.


Florid radiological appearance of megakaryoblastic leukaemia--an aid to earlier diagnosis.
Accurate diagnosis of leukaemia is essential to enable appropriate treatment. The diagnosis is made by cytological and cytochemical techniques but radiology may, however, serve a very useful role in first suggesting a diagnosis. We present here four cases of megakaryoblastic leukaemia (AML FAB classification M7), three of which have strikingly similar unusual radiological changes. A review of the literature suggests this condition has previously been diagnosed as acute or malignant myelofibrosis, and the megakaryoblastic nature of the disease was not recognised. The presence of these radiological features may therefore prompt specific immunocytochemical testing, in a disease that is otherwise difficult to diagnose early.


Microtubule-associated protein 2 appears in axons of cultured dorsal root ganglia and spinal cord neurons after rotavirus infection.
The immunohistochemical distribution of microtubule-associated protein 2 (MAP2), being normally restricted to nerve cell bodies and dendrites, became altered in rat dorsal root ganglia and spinal cord neurons in cultures infected with rhesus rotavirus. MAP2 appeared in axons of both sources of neurons as displayed with monoclonal antibodies to MAP2a + b and MAP2a + b + c at 48 hr post-infection (p.i.). Other cytoskeletal elements, i.e., tau, MAP1, MAP5, neurofilament, actin, and tubulin, did not reveal any alterations in the rotavirus-infected neurons. One of the rotavirus cytosolic proteins, the inner capsid protein vp6, was expressed in axons at 48 hr p.i. simultaneously with the appearance of MAP2, while two other viral proteins, vp4 and NS28, remained in the nerve cell bodies. By quantitative enzyme-linked immunosorbent assay (ELISA) a binding of single-shelled rotaviruses, which express vp6 on their surfaces, to purified MAP2 was found. There was no binding of these viral particles to tau or tubulin proteins. This study indicates that a selective interaction between certain viral and neuronal cytoskeletal proteins can occur and that a non-cytolytic viral infection can cause alterations in the polarized sorting of neuronal proteins.


Enhanced efficiency of a capillary-based biosensor over an optical fiber biosensor for detecting calpastatin.
A capillary-based optical biosensor has been developed to detect calpastatin, an indicator of meat tenderness. Longissimus muscle samples (n=11) were extracted from beef carcasses at 0 and 48h post-mortem. These samples were assayed for calpastatin by traditional laboratory methods and with a newly developed capillary tube biosensor as well as for Warner-Bratzler shear force (WBSF) and crude protein and the responses were compared. Additionally, the response from the capillary-based biosensor was compared to a previously developed optical fiber biosensor. When the 0 and 48h sampling periods were combined, the capillary tube biosensor was moderately accurate in predicting calpastatin activity (R(2)=0.6058). There was less variation in the 0h capillary tube biosensor compared to the 0h pre-column (P=0.006) and post-column optical fiber biosensors (P=0.047), therefore the capillary tube biosensor is a more precise system of measurement. This research further advances the development of a calpastatin biosensor and makes online assessment one step closer to reality.


Prediction model for different progressions of Atherosclerosis in ApoE-/- mice based on lipidomics.
Atherosclerosis (AS) is a progressive disease with a complex pathogenesis which is characterized by dyslipidemia and changes in the vascular wall composition. According to the degree of lesions, atherosclerosis can be divided into four stages: hyperlipidemia, lipid stria, fiber plaque, and atherosclerotic plaque. The present study aimed to establish a prediction model for the different pathological stages of AS based on lipidomics. ApoE-/- mice and C57BL/6 mice fed a normal diet were divided into seven groups according to the feeding time (8, 12, 16, 20, 24, 28, and 32 weeks). The changes in the lipid composition and serum content were detected using ultra-performance liquid chromatography coupled with quadrupole time-of-flight high-definition mass spectrometry (UPLC-Q-TOF/MS). Through the results of serum total cholesterol, triglyceridelow density lipoprotein at each time and HE staining of the head and arm artery, the seven time points of the model group were corresponding to the four courses of atherosclerosis. In accordance with the lipid data of each course of AS and mathematical modeling, this study established a multi-index prediction model of the different processes of AS. Notably, while establishing the model, several indicators were combined with one of four dimension reduction methods, such as principal component logistics regression method, cumulative logistics regression method, Partial least squares-discriminant analysis(PLS-DA), and canonical discriminant analysis (CDA). The error rate of the four methods were 28.5%, 16.22%, 18.24%, and 14.86%, respectively. CDA had the lowest error rate and the best prediction accuracy of the AS different courses for the training and verification sets after 5-fold cross-validation of this model. This study showed that lipidomics combined with mathematical methods could establish a non-invasive and accurate model for the prediction of AS.


Activity standardization of 99mTc by digitizer-based 4πce(LS)-γ and 4πce(PC)-γ coincidence counting.
An aqueous solution of 99mTc was standardized by means of 4πce(LS)-γ and 4πce(PC)-γ coincidence counting. The activity concentration was obtained by efficiency extrapolation relying on four different variation techniques. In both setups, fast digitizers were used for data acquisition. Detected events were recorded in list mode format, and their coincidence relationship was analyzed in the subsequent offline analysis. The combined result was used to participate in an international comparison using the Transfer Instrument of the International Reference System.


Effect of condensation phenomena on potentiometric measurements.
Results of potentiometric analysis, namely those of pH measurements, depend on temperature control of the experimental setup, as it is expressed in the analytical law, the Nernst equation, starting from the primary level, where reference values are conventionally assigned to standard solutions, through the whole traceability chain, down to the service laboratory. Fundamental studies of pH standards, based on the measurement of the potential of an electrochemical cell without transference, known as Harned cell, containing a platinum-hydrogen electrode and a silver-silver chloride reference electrode, refer condensation phenomena on the portions of the cell walls which are not immersed in the thermostatic bath, as one of the major sources of error in the assessment of both the silver-silver chloride electrode standard potential and on pH values. In this work such effect, which is bound to happen due to significant temperature differences between the ambient air temperature and the water bath, has been quantified, presenting an original contribution to the improvement of the quality of potentiometric analysis results. This was possible due to the availability of a climatic cabin "WALKIN" with a temperature control of +/-0.01 degrees C, which permitted that temperature gradients were built between the thermostat water bath (controlled to +/-0.005 degrees C) where cells filled to about 2/3 full were immersed up to 90% of their height, and the surrounding environment.


Elicitation of efficient, protective immune responses by using DNA vaccines against tuberculosis.
DNA vaccination is an effective method for elicitation of strong humoral as well as cellular immune responses. DNA vaccines expressing mycobacterial antigens ESAT-6 (Rv3875), alpha-crystallin (Rv2031c) and superoxide dismutase A (Rv3846) were evaluated for their immune responses in Balb/c mice and protective efficacy in guinea pigs. Immunization of mice with the DNA vaccines expressing superoxide dismutase A and alpha-crystallin resulted in markedly higher levels of IFN-gamma as compared to the levels of IL-10. The DNA vaccine expressing ESAT-6 elicited a mixed Th1/Th2 response. Immunization of guinea pigs with these DNA vaccines and subsequent challenge of animals with Mycobacterium tuberculosis H(37)Rv, showed that DNA vaccine expressing superoxide dismutase imparted the maximum protection as observed by a 50 and 10 folds reduction in bacillary load in spleens and lungs, respectively, in comparison to immunization with vector control.


3,4,9,10-Perylenetetracarboxylic dianhydride functionalized graphene sheet as labels for ultrasensitive electrochemiluminescent detection of thrombin.
A novel tracer, 3,4,9,10-perylenetetracarboxylic dianhydride (PTCDA) functionalized graphene sheet (GS) composite (GS-TCDA), is employed to label the secondary anti-thrombin aptamer (TBA) to construct an ultrasensitive electrochemiluminescent sandwich-type aptasensor. The GS provided large surface area for loading abundant PTCDA and TBA with good stability and biocompatibility. Because of the excellent electroconductivity of GS and the desirable optical properties of PTCDA, the as-formed Apt II bioconjugate considerably amplified the electrochmiluminescence (ECL) signal of peroxydisulfate (S(2)O(8)(2-)) and worked as the desirable label for Apt II. On the basis of the considerably amplified ECL signal and sandwich format, an extremely wide range from 1 fM to 1 nM with an ultralow detection limit of 0.33 fM for thrombin was obtained. Additionally, the selectivity and stability of the proposed aptasensor were also excellent. Thus, this procedure has great promise for detection of thrombin present at ultra-trace levels during early stage of diseases.


A new method for the cytochemical staining of cells immobilized in agarose.
A quick, simple and inexpensive technique utilizing an isotonic matrix of semi-solid agarose for immobilizing channel catfish (Ictalurus punctatus) peripheral blood leucocytes onto microscope slides before cytochemical staining is described. The technique was demonstrated to be superior to the more conventional means of immobilizing cells for procedures requiring either fixed (Sudan Black B and nonspecific esterase stains) or viable (Nitro-Blue Tetrazolium stain) cells. Although the agarose immobilization technique was developed with channel catfish leucocytes, its utility with other fragile cell populations seems likely.


The Covid-19 pandemic as an accelerator of economic worries and labor-related mental health polarization in Germany? A longitudinal interacted mediation analysis with a difference-in-difference comparison.
Labor-related mental health polarization refers to exposure to low-paid employment and unemployment decreasing mental health. Previous research identified economic worries as a key mediator. Against this background, the Covid-19 pandemic is often assumed to have accelerated already existing processes and affected vulnerable populations the most. Our study sought to investigate whether the Covid-19 pandemic accelerated the mediation by economic worries between employment type and mental health. Using the German Socioeconomic Panel (GSOEP) from 2016 onwards, we created a pre-Covid-19 sample (N = 8266) and a per-Covid-19 sample (N = 7294), with each having a t0 wave (2016/2018) and a t1 wave (2018/2020). We applied the mediational g-formula for longitudinal mediation with exposure-mediator (XM) interaction between employment type (X) and economic worries (M). We decomposed the total effect into a direct, indirect, and interacted effect of employment on mental health and provided a difference-in-difference comparison of the effects. During the Covid-19 pandemic, economic worries increased, and mental health decreased. However, the mediation by economic worries reduced by approx. 18.0% (e.g., from 35.0% to 28.9%). A decreased indirect effect caused the reduction in mediation, while the direct and interacted effect remained rather stable. We also found stark gender differences towards males having a higher total effect but a lower mediated effect during the Covid-19 pandemic. Our results highlight that mediators competing to economic worries must have emerged during the Covid-19 pandemic. Such mediators could be the risk of infection at the workplace, the possibility of remote work, and gender-specific mediators. Our study is also the first to extend the mediational g-formula with the difference-in-difference approach. It can be used as a blueprint for researchers interested in evaluating the impact of events, such as the Covid-19 pandemic, on preexisting processes.


Object relations and interpersonal problems in sexually abused female patients: an empirical study with the SCORS and the IIP.
In this study, we examined how retrospective reports of experiencing traumatic sexual abuse in childhood relates to both the development of self-representations and object representations and the occurrence of interpersonal problems. A total of 30 psychosomatic female patients who reported sexual abuse in childhood were compared with a corresponding number of eating-disordered patients and a nonclinical control group. The object relations technique (ORT; Phillipson, 1955), evaluated using the Social Cognition and Object Relations Scale (SCORS; Westen, 1985, 1991b), and the Inventory of Interpersonal Problems (Horowitz, Rosenberg, Baer, & Ureno, 1988) were used to measure the groups. The patients reporting sexual abuse achieved significantly lower scores in the cognitive scales of the SCORS; in the affective scales, they differed from the control group but not from the patients with an eating disorder. Concerning interpersonal problems, the patients reporting childhood sexual abuse reported interpersonal conflicts more frequently. The results of the study support the influence of traumatic sexual abuse on the formation of self-representations and object representations and on the occurrence of interpersonal conflicts.


Treatment of hypertension with the aid of beta-adrenergic blocking drugs.
Beta-adrenergic blocking drugs have become a recognized addition to the hypotensive agents. This article reviews the current beta-blocking agents, with regard to their usage, side-effects and role in the treatment of hypertension.


Synaptonemal complex analysis of X-7 translocations in male mice. I. R3 and R5 quadrivalents.
The synaptonemal complexes of surface-spread spermatocytes of mice heterozygous for one of two reciprocal translations (R3 and R5) between the X and chromosome 7 have been examined by light and electron microscopy (EM). The break points of R3 were determined to be at 70% of chromosome 7, as measured from the centromere, and at 22% of the X. Translocation quadrivalents were formed almost exclusively. The break points of R5 were at 21% of chromosome 7 as measured from the centromere, and at 83% of the X. There was little indication that the break in the X interfered with sex-chromosome synapsis between the 7X and Y. Univalent Y's were not observed in R3, and only seldom observed (8-14%) in R5. However, in contrast to R3, R5 formed quadrivalents relatively rarely (20% in the EM study of 100 nuclei), and heteromorphic bivalents of 7X-Y and X7-7 quite frequently (72%). Possible causes of this high bivalent frequency are discussed. Light-microscope (LM) analysis alone was found to be inadequate for interpreting synaptic configurations (quadrivalents vs. bivalents) in R5. The LM analysis was further complicated by the occurrence of nonhomologous synapsis in the heteromorphic bivalents of R5, a phenomenon easily recognized and interpreted in the EM portion of the study.


Unexpected, complete recovery after emergent thoracic endovascular aortic repair for inoperable type A aortic dissection.
Patients presenting with a Stanford type A acute aortic dissection require immediate surgical treatment; however, up to 30% of patients are deemed inoperable. Here we describe a case of a patient with a complicated type A acute aortic dissection presenting with a severe impact of brain malperfusion. In contrast with open surgery, an emergent thoracic endovascular aortic repair was performed with a Gore cTAG 45 × 150 mm graft and an additional chimney graft Advanta V12 7 × 59 mm graft for the brachiocephalic trunk. After early extubation, unexpected complete neurological recovery was observed. A follow-up computed tomography scan demonstrated complete remodeling of the ascending aorta. This report underlines the potential of thoracic endovascular aortic repair as an alternative for immediate open surgical repair in case of high-risk or inoperable patients.


Neoadjuvant targeted molecular therapies in patients undergoing nephrectomy and inferior vena cava thrombectomy: is it useful?
To assess the effect of neoadjuvant targeted molecular therapies (TMTs) on size and level of inferior vena cava tumor thrombi and to evaluate their impact on surgical management. We retrospectively analyzed the data of 14 patients treated for a clear cell renal cell carcinoma with inferior vena cava thrombi by neoadjuvant TMT before nephrectomy. Clinical, pathological and perioperative data were gathered retrospectively at each institution. The primitive tumor size and the thrombus size were defined by computed tomography before TMT. The tumor thrombus level was defined according to the Novick's classification. Before TMT, thrombus level was staged I for 1 (7%), II for 10 (72%) and III (21%) for 3 patients. First-line therapy was sunitinib in 11 cases and sorafenib in 3 cases. Median therapy duration was two cycles (1-5). Three patients experienced major adverse effects (grade III) during TMT. Following TMT, 6 (43%) patients had a measurable decrease, 6 (43%) had no change, and 2 (14%) had an increase in the thrombus. One patient (7%) had a downstage of thrombus level, 12 (85%) had stable thrombi, and 1 (7%) had an upstage. Regarding primary tumor, 7 (50%), 5 (36%) and 2 (14%) patients had a decrease, stabilization and an increase in tumor size, respectively. Neoadjuvant TMT appears to have limited effects on renal tumor thrombi. This retrospective study failed to demonstrate a significant impact of neoadjuvant TMT on surgical management of clear cell renal cell carcinoma with inferior vena cava tumor thrombi.


Evaluating Reprogramming Efficiency and Pluripotency of the Established Human iPSCS by Pluripotency Markers.
The pluripotency of human induced pluripotent stem cells (HiPSCs) cannot be tested strictly in a similar way as we can do for the mouse ones because of ethical restrictions. One common and initial approach to prove the pluripotency of an established human iPSC line is to demonstrate expression of a set of established surface and intracellular pluripotency markers. This chapter provides procedures of immunocytochemistry of the established HiPSC lines for a set of the signature intracellular pluripotency proteins, OCT4, SOX2, NANOG, and LIN28. We also describe cell phenotyping by flow cytometry for the five established human pluripotency surface markers, SSEA3, SSEA4, TRA-1-60, TRA-1-81, and TRA2-49 (ALP). Numbers of ALP+ and TRA-1-60+ colonies are the most widely used parameters for evaluation of human iPSC reprogramming efficiency. Therefore, this chapter also provides detailed steps for substrate colorimetric reaction of the ALP activity, as well as the TRA-1-60 staining, of the iPSC colonies in the reprogramming population.


Decreased platelet miR-223 expression is associated with high on-clopidogrel platelet reactivity.
We aimed to investigate the relationship between platelet microRNA (miR-223 and miR-96) expression and clopidogrel responsiveness in patients with coronary heart disease (CHD). A total of 33 consecutive non-diabetic CHD patients scheduled for percutaneous coronary intervention were enrolled. Platelet reactivity after clopidogrel loading dose (300 mg) was determined by two methods [platelet reactivity index (PRI), measured by vasodilator-stimulated phosphoprotein (VASP) phosphorylation flow cytometry and ADP-induced platelet aggregation (PAG), measured by light transmission aggregometry]. Total platelet RNA was isolated from purified platelets (CD45 magnetic bead negative selection) to quantify miR-223 and miR-96 expression by real-time PCR. All subjects were dichotomized according to PRI medians (normal-responders: PRI < 56.5%, n = 17 and low-responders: PRI > 56.5%, n = 16) and PAG medians (normal-responders: PAG < 43%, n = 17 and low-responders: PAG > 43%, n = 16). Compared with PRI-determined normal-responders, miR-223 expression, but not miR-96, was significantly decreased in low-responders (P = 0.037). No differential expression of miR-223 and miR-96 was observed via PAG determination between normal- and low-responders. In addition, miR-223 expression, but not miR96, was statistically correlated with PRI (Spearman r = -0.403, P = 0.020). Stepwise binary logistic regression analysis revealed that among factors that potentially influence platelet reactivity (CYP2C19*2 loss-of-function genotypes, use of calcium channel blockers/proton-pump inhibitors, age, obesity, smoking and platelet microRNAs), decreased miR-223 expression was the only independent predictor associated with the presence of PRI-determined low responders to clopidogrel (OR 0.189, 95% CI 0.043 to 0.836, P = 0.028). The present work identifies decreased platelet miR-223 expression as a novel mechanism involved in blunted platelet response to clopidogrel in a Chinese population.


Sports Medicine and Arthroscopic Treatment of the Foot and Ankle: What Is New and Current in Singapore.
Foot and ankle abnormalities are common in Singapore because of the compulsory conscription, the slipper-wearing culture, and the promotion of healthy living through exercise. The rapidly aging population, lack of elite sportsmen, and social and cultural norms pose unique challenges to foot and ankle surgery. Orthopedic surgery in Singapore has progressed because of the good infrastructure and modern practices executed by fellowship-trained surgeons. Evolving local practices are polarized by practice trends emulated from North America and Europe. The small community of foot and ankle surgeons currently practicing in Singapore allows for easier communication, corroborative educational events, and research initiatives.


Serum immunosuppressive acidic protein (IAP) in patients with malignant head and neck tumors.
Measurements were carried out on the serum immunosuppressive acidic protein (IAP) level of patients with malignant head and neck tumors. The serum IAP value in patients with malignancies was significantly higher than that in healthy subjects and those with benign tumors. Following treatment, in cases with favorable progress the high IAP values prior to treatment showed a decline, becoming normal. Whereas in cases showing poor progress, there was a delay in reverting to normal values and some were observed to retain the same high values.


AlphaFold version 2.0 elucidates the binding mechanism between VIPR2 and KS-133, and reveals an S-S bond (Cys25-Cys192) formation of functional significance for VIPR2.
The vasoactive intestinal peptide receptor 2 (VIPR2) has attracted attention as a drug target for the treatment of mental disorders, cancer, and immune diseases. In 2021, we identified the peptide KS-133 as a VIPR2-selective antagonist. In this study, we aimed to elucidate the binding mechanism between VIPR2 and KS-133. To this end, VIPR2/KS-133 and VIPR2/vasoactive intestinal peptide (VIP) complex models were constructed through AlphaFold version 2.0 and molecular dynamic simulations. Our models revealed that: (i) both KS-133 and VIP have helical structures, (ii) the interaction residues on VIPR2 for both peptides are similar, and (iii) the orientation of their helices upon their binding to VIPR2 are different by ∼45°. Interestingly, in the process of constructing the aforementioned models, an S-S bond formation between Cys25 and Cys192 of the human VIPR2 was identified. Although these two Cys residues are highly conserved among species (i.e., corresponding to Cys24 and Cys191 in the mouse), no previous reports regarding this S-S bond formation exist. In order to clarify the potential role of this S-S bond in the VIPR2 has functional consequences, a cell line expressing the mouse VIPR2(Cys24Ala, Cys191Ala) was generated. During the VIP stimulation of this cell line, the phosphorylation of AKT (a downstream signal marker of VIPR2) was found to be significantly attenuated, thereby suggesting that the S-S bond has a functional significance for VIPR2. Our study not only elucidates the VIPR2-binding mechanism of KS-133 for the first time, but also provides new insights into the structural biology of VIPR2.


Total plasma l-tryptophan, free l-tryptophan and competing amino acid levels in a homicidal male adolescent with conduct disorder.
Several studies have shown abnormal findings in human serotonin metabolism, such as increased total plasma l-tryptophan and free l-tryptophan levels among habitually violent antisocial offenders. It is not clear if these increased l-tryptophan levels are associated with adult antisocial personality disorder (ASP) or history of substance abuse, or if these levels are already present in adolescent subjects with conduct disorder (CD). Total plasma and free l-tryptophan and competing amino acids (CAAs) were measured in a 15-year-old adolescent offender, who was convicted for two homicides, and in 10 healthy male controls of similar age and body mass index (BMI). In the juvenile offender, plasma total l-tryptophan/CAA was 84% and free l-tryptophan/CAA 143% higher than average mean among controls. From this very aggressive boy with CD, findings of free l- and total l-tryptophan/CAA values were similar to those of habitually violent adult ASP offenders. As severe CDs in adolescence tend to develop into adults with ASP, increased l-tryptophan/CAA and free l-tryptophan/CAA values may serve as early indicators for the development of habitually violent adult offenders.


Evaluating water quality benefits of manureshed management in the Susquehanna River Basin.
Manureshed management guides the sustainable use of manure resources by matching areas of crop demand (nutrient sinks) with areas generating livestock manure (nutrient sources). A better understanding of the impacts of manureshed management on water quality within sensitive watersheds is needed. We quantified the potential water quality benefits of manureshed-oriented management through scenario-based analyses in the Susquehanna River Basin (SRB) using the Soil and Water Assessment Tool. Five manureshed management scenarios were developed and compared with a baseline "business-as-usual" scenario. The baseline assumes manure is less transportable, which means some locations have manure application in excess of crop demand. The "watershed nutrient balance" scenarios assume excess manure from surplus locations is transportable and that manure is applied around the SRB based on crop nutrient demand. The "watershed nutrient balance avoiding runoff prone areas" scenarios assume manure is transportable but not applied in vulnerable landscapes of the SRB. Each scenario was evaluated under two application rates considering crop nitrogen demand (N-based) and phosphorus demand (P-based). Phosphorus-based manureshed management was more effective in water quality improvements than N-based management. Phosphorus-based nutrient balance scenarios simulated 3 and 25% reduction in total N (TN) and total P (TP), respectively, from the baseline scenario at the watershed outlet. The N- and P-based scenarios avoiding runoff prone areas simulated 3 and 6% reduction in TN loss and 4 and 25.2% reduction in TP loss, respectively, from the baseline. Overall, the manureshed management scenarios were more effective in improving the quality of local streams in livestock-intensive regions than at the watershed outlet.


Protein rotation and chromophore orientation in reconstituted bacteriorhodopsin vesicles.
Bacteriorhodopsin has been reconstituted into lipid vesicles with dipalmitoyl and dimyristoyl phosphatidylcholine. Circular dichroism (CD) measurements show that the proteins are in a monomeric state above the main lipid phase transition temperature (Tc), 41 and 23 degrees C for dipalmitoyl and dimyristoyl phosphatidylcholine, respectively. Below Tc, the CD spectrum is the same as that found for the purple membrane. The latter result implies that the orientation of the chromophore at these temperatures is most likely the same as in the purple membrane (70 degrees +/- 5 degrees from the normal to the membrane plane). Transient dichroism measurements show that below Tc the proteins are immobile, while above this temperature protein rotation around an axis normal to the plane of the membrane is occurring. In addition, from the data the angle of the chromophore for the rotating proteins with respect to the rotational diffusion axis can be calculated. This angle is found to be 30 degrees +/- 3 degrees and 29 degrees +/- 4 degrees in dimyristoyl phosphatidylcholine and dipalmitoyl phosphatidylcholine, respectively. This is considerably smaller than the value of 70 degrees +/- 5 degrees for the natural biomembrane. A reversible reorientation of the chromophore above and below the respective main Tc transition temperature could explain the change of angle observed provided that all the molecules rotate above Tc.


Behavioral and biofeedback therapy for a functionally impaired musician: a case report.
A performing musician with severe occupational palsy accompanied by obsessions and compulsions was treated by an in vivo temperature biofeedback exercise. The treatment appeared highly successful in removing both the palsy and the obsessions, with improvement seen after six 1/2-day sessions and persisting for 4 years after the termination of treatment. This is the second reported case of functional palsy in a performing musician successfully treated by applying feedback apparatus directly to the affected part of the body. Occupational palsy, traditionally thought to be highly resistant to treatment, seems surprisingly amenable to in vivo biofeedback.


Increasing age is not associated with toxicity leading to discontinuation of treatment in patients with urothelial non-muscle-invasive bladder cancer randomised to receive 3 years of maintenance bacille Calmette-Guérin: results from European Organisation for Research and Treatment of Cancer Genito-Urinary Group study 30911.
To determine the relationship of age to side-effects leading to discontinuation of treatment in patients with stage Ta-T1 non-muscle-invasive bladder cancer (NMIBC) treated with maintenance bacille Calmette-Guérin (BCG). We evaluated toxicity for 487 eligible patients with intermediate- or high-risk Ta-T1 (without carcinoma in situ) NMIBC randomised to receive 3 years of maintenance BCG therapy (247 BCG alone and 240 BCG + isoniazid) in European Organisation for Research and Treatment of Cancer Genito-Urinary Group trial 30911. The percentage of patients who stopped for toxicity and the number of treatment cycles that they received were compared in four age groups, ≤60, 61-70, 71-75 and >75 years, using the Mantel-Haenszel chi-square test for trend. The percentage of patients stopping BCG for toxicity was 17.9% in patients aged ≤60 years, 21.9% in patients aged 61-70 years, 22.9% in patients aged 71-75 years, and 16.4% in patients aged >75 years (P = 0.90). For both systemic and local side-effects, there was likewise no significant difference. In patients with intermediate- and high-risk Ta-T1 NMIBC treated with BCG, no differences in toxicity as a reason for stopping treatment were detected based on patient age.


Analysis of 16 autosomal STRs and 17 Y-STRs in an indigenous Maya population from Guatemala.
The aim of this study was to contribute new data on autosomal STR and Y-STR markers of the Mayas from Guatemala in order to improve available databases of forensic interest. We analyzed 16 autosomal STR markers in a population sample of 155 indigenous Maya and 17 Y-chromosomal STR markers in the 100 males of the sample. Deviations from Hardy-Weinberg equilibrium and linkage disequilibrium between autosomal STR markers were not observed at any loci. The combined power of exclusion was estimated as 99.9991% and the combined power of discrimination was >99.999999999999%. Haplotype diversity of Y-STRs was calculated as 0.9984 ± 0.0018 and analysis of pairwise genetic distances (Rst) supported the Native American background of the population.


Age at diagnosis, obesity, smoking, and molecular subtypes in muscle-invasive bladder cancer.
Heterogeneity of muscle-invasive bladder cancer (MIBC) has been characterized using whole-genome mRNA expression data, showing distinct molecular and clinicopathological characteristics by subtypes. However, associations between risk factors and molecular subtypes have not been reported. Four previously published schemes were used to categorize molecular subtypes in 372 MIBC patients from the Cancer Genome Atlas (TCGA). Data on gene expression (RNA-seq), demographic, and clinicopathological characteristics were retrieved through TCGA data portal. Polytomous logistic regression was used to estimate the associations of subtypes by different schemes with age at diagnosis, obesity, and smoking. While some quantitative variation was evident, distinct molecular subtype schemes showed considerable consistency in the association with the risk factors. Generally, compared to patients with luminal-like tumors, patients with basal-like subtypes were more likely to be older (OR75 + yrs vs. <60 years range = 1.32-2.89), obese (ORobese vs. normal range = 1.30-3.05), and to start smoking at early age (OR<18 years vs. 25+ years range = 1.11-4.57). Different molecular subtypes of MIBC may have distinct risk profiles. Large population-based studies with detailed information on bladder cancer risk factors are needed to further define etiologic heterogeneity for bladder cancer.


The endogenous factors affecting the detection of serum SARS-CoV-2 IgG/IgM antibodies by ELISA.
To investigate endogenous interference factors of the detection results of novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) IgM/IgG. Enzyme-linked immunosorbent assay (ELISA) was used to detect SARS-CoV-2 IgM/IgG in sera of 200 patients without COVID-19 infection, including rheumatoid factor (RF) positive group, antinuclear antibody (ANA) positive group, pregnant women group, and normal senior group, with 50 in each group and 100 normal controls. The level of SARS-CoV-2 IgG in pregnant women was significantly higher than that in the normal control group (p = 0.000), but there was no significant difference between other groups. The levels of SARS-CoV-2 IgM in the pregnant women group, normal senior group, ANA positive group, and RF positive group were significantly higher than that in the normal control group (p < 0.05), with significant higher false-positive rates in these groups (p = 0.036, p = 0.004, p = 0.000, vs. normal control group). Serum RF caused SARS-CoV-2 IgM false-positive in a concentration-dependent manner, especially when its concentration was higher than 110.25 IU/L, and the urea dissociation test can turn the false positive to negative. ANA, normal seniors, pregnant women, and RF can lead to false-positive reactivity of SARS-CoV-2 IgM and/or IgG detected using ELISA. These factors should be considered when SARS-CoV-2 IgM or IgG detection is positive, false positive samples caused by RF positive can be used for urea dissociation test.


Experience, complications and prognostic factors of the ileoanal pouch in ulcerative colitis: An observational study.
Ileoanal pouch following restorative proctocolectomy is the treatment for ulcerative colitis after failed medical treatment. Our main aim was to evaluate early and late morbidity associated with restorative proctocolectomy. The secondary aim was to assess risk factors for pouch failure. A retrospective, observational, single-center study was performed. Patients who had undergone restorative proctocolectomy for a preoperative diagnosis of ulcerative colitis from 1983-2015 were included. Early (<30 days) and late (>30 days) adverse events were analyzed. Pouch failure was defined as the need for pouch excision or when ileostomy closure could not be performed. Univariate and multivariate analyses were performed to assess pouch failure risk factors. The study included 139 patients. One patient subsequently died in the early postoperative period. Mean follow-up was 23 years. Manual anastomoses were performed in 54 patients (39%). Early adverse events were found in 44 patients (32%), 15 of which (11%) had anastomotic fistula. Late adverse events were found in 90 patients (65%), and pouch-related fistulae (29%) were the most commonly found in this group. Pouch failure was identified in 42 patients (32%). In the multivariate analysis, age >50 years (p<0.01; HR: 5.55), handsewn anastomosis (p<0.01; HR: 3.78), pouch-vaginal (p=0.02; HR: 2.86), pelvic (p<0.01; HR: 5.17) and cutaneous p=0.01; HR: 3.01) fistulae were the main pouch failure risk factors. Restorative proctocolectomy for a preoperative diagnosis of ulcerative colitis has high morbidity rates. Long-term outcomes could be improved if risk factors for failure are avoided.


Coronary artery bypass graft surgery complications: A review for emergency clinicians.
Coronary artery bypass graft (CABG) surgery remains a high-risk procedure, and many patients require emergency department (ED) management for complications after surgery. This narrative review provides an evidence-based summary of the current data for the emergency medicine evaluation and management of post-CABG surgery complications. While there has been a recent decline in all cardiac revascularization procedures, there remains over 200,000 CABG surgeries performed in the United States annually, with up to 14% of these patients presenting to the ED within 30 days of discharge with post-operative complications. Risk factors for perioperative mortality and morbidity after CABG surgery can be divided into three categories: patient characteristics, clinician characteristics, and postoperative factors. Emergency physicians will be faced with several postoperative complications, including sternal wound infections, pneumonia, thromboembolic phenomena, graft failure, atrial fibrillation, pulmonary hypertension, pericardial effusion, strokes, renal injury, gastrointestinal insults, and hemodynamic instability. Critical patients should be evaluated in the resuscitation bay, and consultation with the primary surgical team is needed, which improves patient outcomes. This review provides several guiding principles for management of acute complications. Understanding these complications and an approach to the management of hemodynamic instability is essential to optimizing patient care. Postoperative complications of CABG surgery can result in significant morbidity and mortality. Physicians must rapidly diagnose these conditions while evaluating for other diseases. Early surgical consultation is imperative, as is optimizing the patient's hemodynamics, including preload, heart rate, cardiac rhythm, contractility, and afterload.


Atypical chest pain.
The initial approach to patients with a chief complaint of chest pain is to rule out myocardial ischemia. There are, however, other life-threatening causes of chest pain, including pulmonary emobilism and aortic dissection among many others. This article reviews several of these disease processes.


Maximal oxygen uptake in Chilean workers of normal nutritional status.
Maximum oxygen uptake (VO2max) was measured directly and predicted from cardiac frequency measurements in 54 healthy Chilean industrial workers aged 20 to 55 years, together with assessment of their dietary intake, body composition and blood chemistry. Measurement of VO2 was performed on a motor-driven treadmill. The predicted VO2max was obtained using a cycle ergometer by two methods: 1) the Astrand-Ryhming nomogram and 2) the linear relationship between "steady state" heart rate (HR) and submaximum work, with subsequent extrapolation to "maximum" heart rate. Extrapolation of the HR/load regression line to 170 bpm permitted determination of the physical working capacity at 170 bpm (W170). VO2max for the 20-29 year group (Group I) averaged 3624 ml.min-1 and decreased to 3066 ml.min-1 in the 50-55 year group (Group IV). Lower values were obtained using the Astrand-Ryhming nomogram and HR/load regression (-15% and -9% respectively). W170 was also affected by age (Group I: 190.6 W and Group IV: 158.5 W). No significant correlation were found between VO2max and plasma variables, with the exception of cholesterol (r = 0.59). On the contrary, anthropometric variables showed significant correlations with VO2max, which permitted the prediction of VO2max using multiple regression equations. The two best correlations were: 1. VO2max = 0.800 - 0.0225.(A) +0.0189.(W)+1.26.(H) (r = 0.87; p less than 0.001) 2. VO2max = 0.996 - 0.0176.(A) + 0.025.(W) + 0.838.(H) + 0.0255.(LBM) (r = 0.88; p less than 0.001) where A = years of age; W = body weight in kg; H = height in m and LBM = lean body mass in kg.


Effects of theophylline and dibutyryl cyclic adenosine monophosphate on the membrane potential of mouse pancreatic beta-cells.
The effects of theophylline and dibutyryl cyclic AMP on the membrane potential of mouse beta-cells were studied with micro-electrodes. They were compared to their effects on insulin release by perifused mouse islets. In 3 mM-glucose, theophylline (10 mM) depolarized the beta-cell membrane and stimulated insulin release, but did not induce electrical activity. Dibutyryl cyclic AMP (1 mM) was without effect. In 7 mM-glucose, theophylline (0.5-2 mM) and dibutyryl cyclic AMP (1 mM) slightly depolarized the beta-cell membrane, induced electrical activity in otherwise silent cells and increased insulin release. A higher concentration of theophylline (10 mM) hyperpolarized the beta-cell membrane, did not induce electrical activity, but also stimulated insulin release. In 10 mM-glucose, the membrane potential of beta-cells exhibited repetitive slow waves with bursts of spikes on the plateau. Under steady state, these slow waves were differently affected by low or high concentrations of theophylline. At 0.5-2 mM, theophylline shortened the intervals, lengthened the slow waves and slightly increased their frequency. On the other hand, 10 mM-theophylline markedly decreased the duration of both intervals and slow waves, and increased their frequency. The effects of 1 mM-dibutyryl cyclic AMP were similar to those of 2 mM-theophylline. With 2-10 mM-theophylline, two other effects were also observed: a transient hyperpolarization with suppression of electrical activity immediately after addition of the methylxanthine and an increase in electrical activity upon its withdrawal. Theophylline and dibutyryl cyclic AMP markedly potentiated insulin release induced by 10 mM-glucose. The magnitude of these changes did not correlate well with the importance of the changes in electrical activity. However, with 2-10 mM-theophylline the increase in release was also preceded by an initial transient inhibition, whereas withdrawal of the methylxanthine was accompanied by a further increase. When Ca influx was inhibited by D600, the slow waves were suppressed, the membrane was depolarized to the plateau level and only few spikes were present. Although theophylline markedly increased insulin release under these conditions, it did not affect the membrane potential. Several conclusions can be drawn from this study. Insulin release and electrical activity in beta-cells can be dissociated when intracellular Ca is used to trigger exocytosis. High concentrations of theophylline produce effects unrelated to cyclic AMP.(ABSTRACT TRUNCATED AT 400 WORDS)


Involvement of carbohydrates as multiple low affinity interaction sites in the self-association of the aggregation factor from the marine sponge Microciona prolifera.
Cell aggregation in the marine sponge Microciona prolifera is mediated by a multimillion molecular weight aggregation factor (MAF) and is based on two functional properties, a Ca2+-independent cell binding activity and a Ca2+-dependent factor-factor self-interaction. Monoclonal antibodies were prepared against purified MAF, and one clone was characterized which selectively inhibited the MAF-MAF association activity. Binding of the blocking antibody (Block 1) to MAF demonstrated that this epitope structure was present in 1100 copies per one MAF molecule of Mr = 2 X 10(7). Such blocking antibodies precipitated a small molecular weight protein-free glycan fraction prepared from MAF by Pronase digestion, thus indicating that the highly repetitive epitope is located in the carbohydrate portion of the molecule. Since the inhibitory activity of the Block 1 antibody could only be achieved when most of the sites were occupied by Fab fragments of this antibody, the self-association of MAF seemed to be based on the polyvalency of the carbohydrate determinants. The affinity of the protein-free glycans was very low as shown by the fact that they did not display any measurable self or MAT binding activity in their monomeric form. After cross-linking them with glutaraldehyde into polymers of the size of MAF, however, the self-interaction could be reconstituted. Thus, the MAF-MAF association activity, which is needed for cell aggregation in sponges, seems to be based on multiple low affinity carbohydrate-carbohydrate interactions, which is different from most interactions mediated by adhesion molecules characterized so far.


Radiocurability by targeting tumor necrosis factor-alpha using a bispecific antibody in carcinoembryonic antigen transgenic mice.
Tumor necrosis factor-alpha (TNF-alpha) enhances radiotherapy (RT) killing of tumor cells in vitro and in vivo. To overcome systemic side effects, we used a bispecific antibody (BsAb) directed against carcinoembryonic antigen (CEA) and TNF-alpha to target this cytokine in a CEA-expressing colon carcinoma. We report the evaluation of this strategy in immunocompetent CEA-transgenic mice. The murine CEA-transfected colon carcinoma MC-38 was used for all experiments. In vitro, clonogenic assays were performed after RT alone, TNF-alpha alone, and RT plus TNF-alpha. In vivo, the mice were randomly assigned to treatment groups: control, TNF-alpha, BsAb, BsAb plus TNF-alpha, RT, RT plus TNF-alpha, and RT plus BsAb plus TNF-alpha. Measurements of endogenous TNF-alpha mRNA levels and evaluation of necrosis (histologic evaluation) were assessed per treatment group. In vitro, combined RT plus TNF-alpha resulted in a significant decrease in the survival fraction at 2 Gy compared with RT alone (p < 0.00001). In vivo, we observed a complete response in 5 (50%) of 10, 2 (20%) of 10, 2 (18.2%) of 11, and 0 (0%) of 12 treated mice in the RT plus BsAb plus TNF-alpha, RT plus TNF-alpha, RT alone, and control groups, respectively. This difference was statistically significant when TNF-alpha was targeted with the BsAb (p = 0.03). The addition of exogenous TNF-alpha to RT significantly increased the endogenous TNF-alpha mRNA level, particularly when TNF-alpha was targeted with BsAb (p < 0.01). The percentages of necrotic area were significantly augmented in the RT plus BsAb plus TNF-alpha group. These results suggest that targeting TNF-alpha with the BsAb provokes RT curability in a CEA-expressing digestive tumor syngenic model and could be considered as a solid rationale for clinical trials.


Multiple hyperdontia: Report of an unusual case.
Supernumerary teeth are an infrequent developmental anomaly that can appear in any area of the dental arch and can affect any dental organ. Multiple supernumerary teeth, or hyperdontia, is rare in people with no other associated diseases or syndromes. Conditions commonly associated with hyperdontia include cleft lip and palate, trichorhinophalangeal syndrome, cleidocranial dysplasia, and Gardner's syndrome. A black girl, aged 11 years 8 months, came for consultation; radiographs showed 81 teeth: 18 deciduous, 32 permanent, and 31 supernumerary. The main concern initially was to determine whether she was syndromic, and she was referred to a geneticist. G banding analysis showed pericentric inversion of chromosome 9; the chromosome formula was 46, XX, inv (9) (p13q21). Orthodontic treatment for this patient will be a clinical challenge because of the great number of teeth to be extracted and the alterations in the shapes of the teeth. Treatment goals should be established by a multidisciplinary team, where oral surgeon, orthodontist, periodontist, and prosthodontist come together to solve a medical and dental puzzle, eliminating the pieces that do not fit and searching for new ones to obtain an occlusion that will give the patient physiologic conditions of normality and esthetic satisfaction.


Spinal anatomy in an essential illustrated medical treatise of the late Ottoman era.
The aim of this article is to introduce a great physician of the late Ottoman period and his writings on spinal anatomy. The article presents chapters related to the anatomy of vertebra and the vertebral column in the book of Şanizade Ataullah Mehmed, Mir'āt al-Abdān fı Tashrıḥ-i A'ḍāi'l-Insān (Mirror of the Bodies in the Dissection of the Members of the Human Body). Works of Şanizade are viewed as important landmarks in Turkish medical history. Anatomy education in Turkey can be divided into four historic periods and the second period is called the Şanizade Period (1816-1827). This period leads up to the opening of the first modern medical school in Turkey on March 14, 1827. The most important characteristic of Şanizade's work is that it is the first illustrated anatomy book printed in the Ottoman Age, and the value of the work is shown by Şanizade being one of the originators of medical terms in Ottoman Medicine.


Metastatic ameloblastoma responding to combination chemotherapy: case report and review of the literature.
Ameloblastoma is a rare benign odontogenic tumor with locally aggressive behavior and a high recurrence rate. When metastases occur, which are uncommon, lungs constitute the most frequent site involved. Malignant ameloblastomas are different from ameloblastic carcinomas. Malignant ameloblastomas are tumors considered metastatic despite the appearance of well-differentiated or benign histology, while ameloblastic carcinomas are histologically malignant in both primary and metastatic sites. A 24-year-old Moroccan man presented a malignant ameloblastoma of the mandible. The tumor was entirely resected. Five years later, a local recurrence occurred. Our patient was treated by exclusive radiotherapy with persistence of a residual disease. After two years he developed multiple lung metastases. Our patient received a combination chemotherapy using doxorubicin and cisplatin. Less than 50 cases of ameloblastoma with metastases have been reported. There is still no standard treatment for metastatic ameloblastoma. Only through continuous reporting of such cases will clinicians be able to draw an optimal strategy for management of this pathology.


Multicompartmental pressures in the knee joint during arthroscopy.
It is well known that fast changes of position of the knee joint cause high instantaneous intraarticular pressures. During arthroscopy, rotation and flexion of the leg can create a closed-off compartment, increasing the risk of capsular rupture. To evaluate the risk of compartmentation, a study has been conducted in which the intraarticular pressure in 12 knee joints of three women and nine men was measured with Myopress catheters (ATOS Medical AB, Hörby, Sweden) located in the suprapatellar pouch, and the anteromedial and the posteromedial recesses during arthroscopy. This study showed that a starting pressure of 100 mm Hg causes a significantly higher peak pressure than does 75 mm Hg. The pressure varies simultaneously in all compartments without signs of local compartmentation in any position of the joint within the range of 0-90 degrees of flexion. With a positive pressure in the knee joint, no compartmentation occurs within the suprapatellar pouch or anteromedial posteromedial compartments during an arthroscopic procedure.


Identification of Xanthine Oxidase Inhibitors from Celery Seeds Using Affinity Ultrafiltration-Liquid Chromatography-Mass Spectrometry.
Celery seeds have been used as an effective dietary supplement to manage hyperuricemia and diminish gout recurrence. Xanthine oxidase (XOD), the critical enzyme responsible for uric acid production, represents the most promising target for anti-hyperuricemia in clinical practice. In this study, we aimed to establish a method based on affinity ultrafiltration-liquid chromatography-mass spectrometry (UF-LC-MS) to directly and rapidly identify the bioactive compounds contributing to the XOD-inhibitory effects of celery seed crude extracts. Chemical profiling of celery seed extracts was performed using UPLC-TOF/MS. The structure was elucidated by matching the multistage fragment ion data to the database and publications of high-resolution natural product mass spectrometry. Thirty-two compounds, including fourteen flavonoids and six phenylpeptides, were identified from celery seed extracts. UF-LC-MS showed that luteolin-7-O-apinosyl glucoside, luteolin-7-O-glucoside, luteolin-7-O-malonyl apinoside, luteolin-7-O-6'-malonyl glucoside, luteolin, apigenin, and chrysoeriol were potential binding compounds of XOD. A further enzyme activity assay demonstrated that celery seed extract (IC50 = 1.98 mg/mL), luteolin-7-O-apinosyl glucoside (IC50 = 3140.51 μmol/L), luteolin-7-O-glucoside (IC50 = 975.83 μmol/L), luteolin-7-O-6'-malonyl glucoside (IC50 = 2018.37 μmol/L), luteolin (IC50 = 69.23 μmol/L), apigenin (IC50 = 92.56 μmol/L), and chrysoeriol (IC50 = 40.52 μmol/L) could dose-dependently inhibit XOD activities. This study highlighted UF-LC-MS as a useful platform for screening novel XOD inhibitors and revealed the chemical basis of celery seed as an anti-gout dietary supplement.


Effect of Constant Illumination on the Function of the Hypothalamic-Pituitary-Adrenal Axis in Nonhuman Primates.
We studied the effect of constant illumination on the effects of administration of arginine vasopressin (AVP), one of the most important regulators of the key adaptive hypothalamic-pituitary-adrenal (HPA) axis under basal conditions and during stress, as well as on the circadian rhythm of activity of HPA axis and the pineal gland in laboratory primates. In young adult female rhesus monkeys exposed to constant illumination for 7 weeks, the rise in the concentration of ACTH and cortisol in response to administration of AVP was markedly reduced in comparison with both the basal period and with the control group of animals. In addition, a destructive effect of constant lighting on circadian rhythm of cortisol secretion was observed in the absence of significant circadian changes in melatonin secretion. The inhibitory effect of constant illumination on the function of the HPA axis under basal conditions and under conditions of its activation can reduce the body's adaptive abilities.


Dislocation sink annihilating threading dislocations in strain-relaxed Si1-x Ge x layer.
We proposed a dislocation sink technology for achieving Si1-x Ge x multi-bridge-channel field-effect-transistor beyond 5 nm transistor design-rule that essentially needs an almost crystalline-defect-free Si1-x Ge x channel. A generation of a dislocation sink via H+ implantations in a strain-relaxed Si0.7Ge0.3 layer grown on a Si substrate and a following annealing almost annihilate completely misfit and threading dislocations located near the interface between a relaxed Si0.7Ge0.3 layer and a Si substrate. A real-time (continuous heating from room temperature to 600 °C) in situ high-resolution-transmission-electron-microscopy and inverse-fast-Fourier-transform image observation at 1.25 MV acceleration voltage obviously demonstrated the annihilation process between dislocation sinks and remaining misfit and threading dislocations during a thermal annealing, called the [SiI or GeI + V Si or V Ge → Si1-x Ge x ] annihilation process, where SiI, GeI, V Si, and V Ge are interstitial Si, interstitial Ge, Si vacancy, and Ge vacancy, respectively. In particular, the annihilation process efficiency greatly depended on the dose of H+ implantation and annealing temperature; i.e. a maximum annihilation process efficiency achieved at 5 × 1015 atoms cm-2 and 800 °C.


Continuous determination of volatile products in anaerobic fermenters by on-line capillary gas chromatography.
Bio-ethanol and biogas produced during the anaerobic conversion of organic compounds has been a subject of great interest since the oil crisis of the 1970s. In ethanol fermentation and anaerobic treatment of wastewaters, end-product (ethanol) and intermediate-products (short-chain fatty acids, SCFA) cause inhibition that results in reduced process efficiency. Control of these constituents is of utmost importance for bioreactor optimization and process stability. Ethanol and SCFA can be detected with precision by capillary gas chromatography usually conducted in off-line measurements. In this work, an on-line monitoring and controlling system was developed and connected to the fermenter via an auto-sampling equipment, which could perform the feeding, filtration and dilution of the sample and final injection into the gas chromatograph through an automation-based programmed procedure. The sample was continuously pumped from the recycle stream of the bioreactor and treated using a microfiltration unit. The concentrate was returned to the reactor while the permeate was quantitatively mixed with an internal standard solution. The system comprised of a gas chromatograph with the flow cell and one-shot sampler and a PC with the appropriate software. The on-line measurement of ethanol and SCFA, directly from the liquid phase of an ethanol fermenter and a high-rate continuous mode anaerobic digester, was accomplished by gas chromatography. Also, this monitoring and controlling system was proved to be effective in the continuous fermentation of alcohol-free beer.


Mapping of the residues responsible for the negative cooperativity of the receptor-binding region of insulin.
Insulin binding to its receptor leads to negatively cooperative interactions among the receptor sites. Studies with 29 insulin analogues (animal insulins and proinsulin, insulin-like growth factor and chemically modified insulins) which vary 1,000-fold in their affinity for the receptor and in their biological potency, suggest that a discrete invariable region on the surface of the insulin monomer is responsible for including the negative cooperativity. This domain comprises some of the eight carboxy-terminal residues of the B-chain and the A21 asparagine. Burying of this 'cooperative site' in the dimerisation of insulin leads to a loss of negative cooperativity. A revised mapping of the insulin molecule is proposed, featuring distinct bioactive and cooperative sites.


The water mite family Hygrobatidae Koch in Australia. The genera Aspidiobatella Cook, Australorivacarus Viets, Gondwanabates Imamura and Rhynchaustrobates Cook (Acari: Hydrachnidia).
The following eight new species are described from Australia: Gondwanabates crassipalpis, G. flavus, G. spatulifer, Rhynchaustrobates bipartitus, R. sclerosus, R. truusae, R. uniscutatus and R. victorianus. Numerous new records are given of the following hygrobatid genera in Australia: Aspidiobatella Cook, 1986, Australoricacarus K.O. Viets, 1978, Gondwanabates Imamura, 1984 and Rhynchaustrobates Cook, 1986. The males of Aspidiobatella smola Cook and Gondwanabates hopkinsi Imamura and the female of Gondwanabates crassipes Cook are described for the first time.


Basic Bayesian methods.
In this chapter, we introduce the basics of Bayesian data analysis. The key ingredients to a Bayesian analysis are the likelihood function, which reflects information about the parameters contained in the data, and the prior distribution, which quantifies what is known about the parameters before observing data. The prior distribution and likelihood can be easily combined to from the posterior distribution, which represents total knowledge about the parameters after the data have been observed. Simple summaries of this distribution can be used to isolate quantities of interest and ultimately to draw substantive conclusions. We illustrate each of these steps of a typical Bayesian analysis using three biomedical examples and briefly discuss more advanced topics, including prediction, Monte Carlo computational methods, and multilevel models.


Noninvasive multimodal imaging in diagnosing polypoidal choroidal vasculopathy.
To investigate the diagnostic accuracy of noninvasive multimodal imaging methods in diagnosing polypoidal choroidal vasculopathy (PCV) and distinguishing PCV from typical neovascular age-related macular degeneration (nvAMD). Retrospective study. Imaging features of noninvasive multimodal imaging methods, including fundus photography (FP), B-scan optical coherence tomography (OCT), en face OCT, OCT angiography, and autofluorescence, of 103 eyes with PCV or typical nvAMD were reviewed. Diagnostic strategy was established based on imaging features and was validated in other 105 eyes with PCV or typical nvAMD. Features of subretinal orange nodule on FP, thumb-like PED on OCT, notched PED on OCT, bubble sign on OCT, and Bruch's membrane depression under serosanguinous PED on OCT were more common. When the diagnostic strategy of using at least 2 of 5 features was performed, there is 0.88 sensitivity and 0.92 specificity for diagnosing PCV. The results of the validation test further confirmed the diagnostic strategy with 0.94 sensitivity and 0.93 specificity. Noninvasive multimodal imaging, especially FP and B-scan OCT, provide high sensitivity and specificity for diagnosing PCV and distinguishing PCV from typical nvAMD, when at least 2 of 5 suggestive imaging features are present.


Molecular dynamics simulation of a high-affinity antibody-protein complex: the binding site is a mosaic of locally flexible and preorganized rigid regions.
One nanosecond molecular dynamic (MD) simulation of anti-hen egg white lysozyme (HEL) antibody HyHEL63 (HH63) complexed with HEL reveals rigid and flexible regions of the HH63 binding site. Fifty conformations, extracted from the MD trajectory at regular time intervals were superimposed on HH63-HEL X-ray crystal structure, and the root mean squared deviations (RMSDs) and deviations in Calpha atom positions between the X-ray structure and the MD conformer were measured. Residue positions showing the large deviations in both light chain and heavy chain of the antibody were same in all the MD conformers. The residue positions showing smallest deviations were same for all the conformers in the case of light chain, whereas relatively variable in the heavy chain. Positions of large and small deviations fell in the complementarity determining regions (CDRs), for both heavy and light chains. The larger deviations were in CDR-2 of light and CDR-1 of heavy chain. Smaller deviations were in CDR-3 of light and CDR-2 and CDR-3 of heavy chains. The large and small deviating regions highlight flexible and rigid regions of HH63 binding site and suggest a mosaic binding mechanism, including both "induced fit" and preconfigured "lock-and-key" type of binding. Combined "induced fit" and "lock-and-key" binding would be a better definition for the formation of large complexes, which bury larger surface area on binding, as in the case of antibody-HEL complex. We further show that flexible regions, comprising mostly charged and polar residues, form intermolecular interactions with HEL, whereas rigid regions do not. Electrostatic complementarity between HH63 and HEL also imply optimized binding affinity. Flexible and rigid regions of a high-affinity antibody are selected during the affinity maturation of the antibody and have specific functional significance. The functional importance of local inherently flexible regions is to establish intermolecular contacts or they play a key role in molecular recognition, whereas local rigid regions provide the structural framework.


Indigenous cellulolytic aerobic and facultative anaerobic bacterial community enhanced the composting of rice straw and chicken manure with biochar addition.
Microbial degradation of organic matters is crucial during the composting process. In this study, the enhancement of the composting of rice straw and chicken manure with biochar was evaluated by investigating the indigenous cellulolytic bacterial community structure during the composting process. Compared with control treatment, composting with biochar recorded higher temperature (74 °C), longer thermophilic phase (> 50 °C for 18 days) and reduced carbon (19%) with considerable micro- and macronutrients content. The bacterial community succession showed that composting with biochar was dominated by the cellulolytic Thermobifida and Nocardiopsis genera, which play an important role in lignocellulose degradation. Twenty-three cellulolytic bacterial strains were successfully isolated at different phases of the composting with biochar. The 16S rRNA gene sequencing similarity showed that they were related to Bacillus licheniformis, Bacillus subtilis, Bacillus aerius, and Bacillus haynesii, which were known as cellulolytic bacteria and generally involved in lignocellulose degradation. Of these isolated bacteria, Bacillus licheniformis, a facultative anaerobe, was the major bacterial strain isolated and demonstrated higher cellulase activities. The increase in temperature and reduction of carbon during the composting with biochar in this study can thus be attributed to the existence of these cellulolytic bacteria identified.


Intra-prostatic fiducial markers and concurrent androgen deprivation.
To document the case of a man with adenocarcinoma of the prostate treated with external beam radiotherapy and concurrent androgen deprivation. The man, who received 79.8 Gy in 42 fractions of radiotherapy over 8.5 weeks using three intra-prostatic gold fiducial markers for on-line set-up correction, started an anti-androgen 2.5 weeks before radiotherapy, on the day of his planning computed tomography, and a gonadotropin-releasing hormone agonist on his first day of radiotherapy. In the sixth week of radiotherapy, the distance between the fiducial markers had diminished: superior to posterior-mid (from 19 to 11 mm), posterior-mid to inferior (from 19 to 15 mm), and superior to inferior (from 31 to 22 mm), so the patient was rescanned. Between the two planning computed tomographies, the prostate volume had decreased from 44.3 to 28.3 cm3 (-36%). Had the planned radiotherapy been delivered to the anatomy of the rescan, the dose to the rectal wall would have exceeded the planned dose-volume histogram constraints. However, with the patient set up to the fiducial markers, the dose-volume histogram constraints for the rectal wall and bladder wall were met throughout the course of radiotherapy. Involution of the prostate owing to concurrent androgen deprivation may cause in-migration of implanted fiducial markers and excessive dose to the rectal wall. With concurrent androgen deprivation, daily on-line set-up correction to fiducial markers can aid in safe dose escalation.