Trough levels and concentration time curves of cyclosporine in patients undergoing renal transplantation.
We determined the AUC of cyclosporine (24 hours) nonspecifically by RIA and specifically by HPLC after an oral and an intravenous dose of cyclosporine in 58 patients undergoing renal transplantation. The RIA/HPLC concentration ratio of cyclosporine changed continuously during the first 12 hours after administration. The ratio was higher after oral than after intravenous administration and varied from patient to patient. The predictive value of trough levels for the corresponding AUCs was better when trough levels were assessed 24 than 12 hours after administration. Trough levels assessed by RIA poorly predicted AUCs measured specifically by HPLC. Therefore if, in the future, therapeutic cyclosporine monitoring has to be improved, trough levels should be assessed 24 hours after the last dose by means of a specific HPLC method.


Phenotypic studies on dopamine receptor subtype and associated signal transduction mutants: insights and challenges from 10 years at the psychopharmacology-molecular biology interface.
Mutants with targeted gene deletion ('knockout') or insertion (transgenic) of D1, D2, D3, D4 and D5 dopamine (DA) receptor subtypes are complemented by an increasing variety of double knockout and transgenic-'knockout' models, together with knockout of critical components of DA receptor signalling cascades such as G alpha(olf)[G gamma7], adenylyl cyclase type 5, PKA [RIIbeta] and DARPP-32. However, it is increasingly recognised that these molecular techniques have a number of inherent limitations. Furthermore, there are poorly understood methodological factors that contribute to inconsistent phenotypic findings between laboratories. This review seeks to document the impact of DA receptor subtype and related transduction mutants on our understanding of the behavioural roles of these entities, primarily at the level of unconditioned psychomotor behaviour. It includes ethologically based and orofacial movement studies in our own laboratories, since these are the only studies to systematically compare each of the D1, D2, D3, D4 and D5 receptor and DARPP-32 signal transduction 'knockouts'. There is a particular emphasis on identifying methodological factors that might influence phenotypic effects and account for inconsistencies. The findings are offered empirically to (1) specify the extent of phenotypic diversity among individual DA receptor subtypes and transduction components and (2) indicate relationships between D1, D2, D3, D4 and D5 receptor subtype proteins, associated G alpha(i)/G alpha(s)/G alpha(olf)[G gamma7]-adenylyl cyclase type 5-PKA [RIIbeta]-DARPP-32 signalling cascades and behaviour. The findings are also offered heuristically as a base for such phenotypic comparisons at additional levels of behaviour so that a yet more complete phenotypic profile might emerge.


Meteorological influences on daily variation and trend of summertime surface ozone over years of 2015-2020: Quantification for cities in the Yangtze River Delta.
We quantify the meteorological influences on daily variations and trends of maximum daily 8-h average ozone (MDA8 O3) concentrations by using multiple linear regression (MLR) and Lindeman, Merenda, and Gold (LMG) approaches. Different from previous region-based studies, we pay special attention to meteorological influences at city scale. Over 2015-2019, daily changes in key meteorological parameters could explain 47%-74% of the observed daily variations in summertime MDA8 O3 concentrations in Yangtze River Delta (YRD) and four cities (Shanghai, Nanjing, Hangzhou, and Hefei), with RH being the top driver. Over years of 2015-2020, daily concentrations of MDA8 O3 obtained from MLR equations (MDA8O3_MLR) of the local cities always had better performance than those of YRD. Compared with the observed daily MDA8 O3 in June-July-August (JJA) over the studied period, daily MDA8O3_MLR of the local cities (of YRD) had correlation coefficients of 0.73 (0.63), 0.75 (0.74), 0.79 (0.78), and 0.76 (0.73) in Shanghai, Nanjing, Hangzhou, and Hefei, respectively, and the MDA8O3_MLR of the local cities (of YRD) captured 54% (17%), 63% (51%), 52% (27%) of the observed O3-polluted days (days with MDA8 O3 concentration exceeding 160 μg m-3) in Shanghai, Nanjing, and Hangzhou, respectively. The meteorologically driven trends (Trend_Met) in MDA8 O3 were calculated using the established MLR equations. Over 2015-2019, the observed trends (Trend_Obs) and Trend_Met in MDA8 O3 were mostly positive in YRD, Nanjing, Hangzhou, and Hefei. In Shanghai, Trend_Obs, Trend_Met, and anthropogenically driven trend (estimated as Trend_Obs minus Trend_Met) of MDA8 O3 in JJA over 2015-2019 were -1.3, +1.0, and -2.3 μg m-3 y-1, respectively, indicating that the emission control measures alleviated O3 pollution in this city. Our results suggest that it is necessary to establish MLR equations at city scale to account for the role of meteorology in the actions of O3 pollution control.


Lipoxygenase/H2O2-catalyzed oxidation of dihdroxyindoles: synthesis of melanin pigments and study of their antioxidant properties.
5,6-Dihydroxyindole (DHI) and 5,6-dihydroxyindole-2-carboxylic acid (DHICA), which are important intermediates in melanogenesis, can be converted into the corresponding melanin pigments by the action of the lipoxygenase/H2O2 system. Kinetic and HPLC analyses indicate that both DHI and DHICA are good substrates for this enzymatic system. Enzyme activity on both substrates was measured in comparison with peroxidase and tyrosinase; the oxidizing behaviour of lipoxygenase is more similar to that of peroxidase rather than that of tyrosinase. The antioxidant properties of DHI- and DHICA-melanins have been investigated in comparison with other kinds of melanins. DHICA-melanin shows a more pronounced antioxidant effect than that of DHI-melanin and this behaviour can be ascribed to the different structure and solubility of the two pigments. The mixed polymer synthesized from DHI and DHICA is the most effective one. Some implications about the possible explanation of the above mentioned behaviour are discussed.


Loss of mammal-specific tectorial membrane component carcinoembryonic antigen cell adhesion molecule 16 (CEACAM16) leads to hearing impairment at low and high frequencies.
The vertebrate-restricted carcinoembryonic antigen gene family evolves extremely rapidly. Among their widely expressed members, the mammal-specific, secreted CEACAM16 is exceptionally well conserved and specifically expressed in the inner ear. To elucidate a potential auditory function, we inactivated murine Ceacam16 by homologous recombination. In young Ceacam16(-/-) mice the hearing threshold for frequencies below 10 kHz and above 22 kHz was raised. This hearing impairment progressed with age. A similar phenotype is observed in hearing-impaired members of Family 1070 with non-syndromic autosomal dominant hearing loss (DFNA4) who carry a missense mutation in CEACAM16. CEACAM16 was found in interdental and Deiters cells and was deposited in the tectorial membrane of the cochlea between postnatal days 12 and 15, when hearing starts in mice. In cochlear sections of Ceacam16(-/-) mice tectorial membranes were significantly more often stretched out as compared with wild-type mice where they were mostly contracted and detached from the outer hair cells. Homotypic cell sorting observed after ectopic cell surface expression of the carboxyl-terminal immunoglobulin variable-like N2 domain of CEACAM16 indicated that CEACAM16 can interact in trans. Furthermore, Western blot analyses of CEACAM16 under reducing and non-reducing conditions demonstrated oligomerization via unpaired cysteines. Taken together, CEACAM16 can probably form higher order structures with other tectorial membrane proteins such as α-tectorin and β-tectorin and influences the physical properties of the tectorial membrane. Evolution of CEACAM16 might have been an important step for the specialization of the mammalian cochlea, allowing hearing over an extended frequency range.


Modeling the three-dimensional structure of RNA.
The limited number of RNA structures determined by X-ray crystallography and NMR spectroscopy compels the use of experimental and theoretical methods that are less precise to obtain information on RNA conformation. RNA flexibility, a consequence of rotational freedom about seven intra- and internucleotide bonds, is unfortunately of such magnitude that these alternate techniques fall short of providing sufficient information to build robust tertiary structures. Various RNA modeling methods, described herein, permit the organization of this structural data to the form of three-dimensional structures. Interactive computer graphics techniques, for example, have generated several useful models. Also, conventional computer algorithms involving the minimization of empirical energy functions, previously limited to small molecules, are giving way to methods able to handle much larger molecules. Modified distance geometry and molecular mechanics algorithms, using simplified "pseudoatom" representations, can generate structures consistent with input data. A constraint satisfaction algorithm combined with discrete representations of nucleotide conformations systematically explores poorly defined regions of a molecule yielding all-atom representations, but requires enough structural constraints to avoid a computational explosion.


Headache as a side effect of combination estrogen-progestin oral contraceptives: a systematic review.
We conducted a 2-part systematic review of published studies to examine the evidence that combination oral contraceptives can aggravate or cause headache. We used trials with a control group to assess headache risk that was attributable to oral contraceptive use and prospective cohort trials to answer important clinical questions about the natural history and treatment response of headache that occurs with oral contraceptive use. Because of differences in study populations, oral contraceptive formulations, trial end points and trial duration, it was not possible to pool data; but the evidence supports several conclusions. There is little indication that oral contraceptives have a clinically important effect on headache activity in most women. Headache that occurs during early cycles of oral contraceptive use tends to improve or disappear with continued use. No evidence supports the common clinical practice of switching oral contraceptives to treat headache; however, manipulating the extent or duration of estrogen withdrawal may provide benefit.


[Diabetes mellitus in automobile transport drivers--incidence, occupational performance].
The authors present prevalence of diabetes mellitus and atherosclerosis risk factors among automobile transport drivers, relationships of those factors with the work peculiarities. Findings are deteriorated operative response system parameters in automobile transport drivers having diabetes mellitus and arterial hypertension.


Quantitative proteomic profiling of tumor cell response to telomere dysfunction using isotope-coded protein labeling (ICPL) reveals interaction network of candidate senescence markers.
Telomerase inhibition causes progressive telomere shortening and cellular senescence, which constitutes a universal barrier to tumor growth and therefore an attractive target for tumor therapy. To expand our previous studies, we investigated the global effects of telomere dysfunction on the proteome of tumor cells in order to find novel senescence biomarkers. Telomerase-deficient HCT-116 cell clones were analyzed by a quantitative proteomic approach using isotope-coded protein labeling (ICPL) and nanoflow-HPLC-MS/MS. Stringent reduction of the extensive proteomic data from this tumor cell model revealed a list of 59 markers including proteins identified in our former studies and a number of novel proteins involved in tumorigenesis and metastasis such as SFN, S100A4, ANXA2, and LGALS1. A loss of the chromatin protein HMGB2 was demonstrated not only in various telomerase-inhibited clones of different tumor cell lines, but also in normal human fibroblasts undergoing replicative senescence and in aging telomerase knockout mice. Impressively, a coherent and dense network of protein-protein interactions for the bulk of the markers and their implementation in signaling pathways involving key regulators for tumorigenesis were revealed. These results have an impact on the understanding of telomere- and senescence-related signal transduction in tumor cells in consideration of the general lack of senescence markers. Induction of cellular senescence constitutes a potent concept for tumor therapy which interferes with immortalization and additional hallmarks of cancer. The application of a powerful quantitative proteomic approach using isotope-coded protein labeling to an approved model for senescence represented by telomerase inhibited tumor cells led to the identification of novel candidate biomarkers for telomere dysfunction and replicative senescence. Thereby, the identified markers not only fit in the context of the investigated processes with a relevance for additional hallmarks of cancer but are also involved in a strong interaction network and integrated in canonical pathways centered around key cancer-relevant proteins. These potential markers alone or in combination will significantly extend the view on telomere-associated signal transduction in tumor cells and contribute to the field of cellular senescence and aging in consideration of the general lack of biomarkers in this regard.


A case series of three patients presenting with isoniazid induced toxicity and N-acetyl transferase 2 gene mutation: A management conundrum for programmatic therapy of tuberculosis in India.
Isoniazid is an essential drug in the management of tuberculosis but there is a high degree of variation in the Indian population's capacity to acetylate or inactivate isoniazid to the inactive metabolite acetyl isoniazid, and they can be distinctly characterized phenotypically as being either slow or rapid inactivators (the concentration of the enzyme being higher in rapid inactivators). Several mutations in the N-acetyl transference 2 (NAT2) gene account for majority of the slow acetylator genotypes in the human population (NAT2*5A, NAT2*5B, and NAT2*6A). Such individuals are at a greater risk of drug-induced adverse reactions due to reduced drug elimination, compared to those possessing the wild type allele. Here we discuss two cases of Isoniazid induced peripheral neuropathy and one case of Isoniazid induced hepatotoxicity confirmed as having heterozygous or homozygous NAT2 gene mutation. In all 3 cases, the presence of NAT2 gene mutation was associated with the adverse events and the adverse events subsided on stopping or reducing the dose of isoniazid. However, due to lack of guideline-based management of adverse events occurring due to isoniazid, the drug was either abruptly stopped or dosage lowered based only on clinical expertise, which could potentially lead to further resistance to Mycobacterium tuberculosis (as occurred in one of our cases). Further studies of the relationship between NAT2 genotypes, isoniazid concentrations and adverse drug events are required to make genotyping a helpful tool for optimizing isoniazid's therapeutic response and minimizing adverse drug reactions, particularly in countries with a high burden of tuberculosis.


Revisiting the surface anatomy of the sciatic nerve in the gluteal region in children using computed tomography.
No anatomy text specifically describes the course of the sciatic nerve (SN) in the gluteal region in children. Anatomical information is largely derived from cadaveric studies of adults, so accurate anatomical information about the location of the SN in children is required. The aim of this study is to assess the surface anatomy of the SN in children using computed tomography (CT). After excluding studies with pelvic pathology, 75 CT scans were analyzed. Three groups were selected for analysis (0-2, 4-6, and 8-10 years). The position of the SN was measured between the posterior superior iliac spine (PSIS) and the ischial tuberosity (IT) and between the IT and the greater trochanter (GT) using 3-dimensional images. In the 0-2 age group, the SN crossed the middle third of a line between the PSIS and the IT in 94% and the GT and the IT in 80% of cases. In the 4-6 age group, the SN crossed the middle third of a line between the PSIS and the IT in 96% and the GT and the IT in 87%. In the 8-10 age group, the SN crossed the middle third of a line between the PSIS and the IT in 100% and the GT and the IT in 71%. The findings indicate that the SN in children is most accurately located in the middle third along a line drawn from the PSIS to the IT and the GT to the IT. Our study is the first to provide anatomical CT data from living children to guide interventions in the gluteal region.


[Syndrome of hemorrhagic shock and encephalopathy in a newborn with generalized hemangiomatosis of the skin].
A case is described of a newborn with generalized haemangiomatosis of the with clinical symptoms and sings resembling the syndrome of haemorrhagic shock and encephalopathy. It seems that this syndrome, described for the first time by Levin et al. in 1983 is of special interest because of its sudden onset, dramatic course, and high mortality.


Ophiopogonin D ameliorates DNCB-induced atopic dermatitis-like lesions in BALB/c mice and TNF-α- inflamed HaCaT cell.
Atopic dermatitis (AD) can occur in both children and adults, and the symptoms include itching and eczema, which in turn cause patients to suffer. Ophiopogonin D (OP-D) is a steroidal glycoside from Radix Ophiopogon japonicus, which is well known as an effective anti-inflammatory herbal medicine in many Asian countries. In this study, we aimed to investigate the anti-inflammatory effects of OP-D, using an AD mouse model and inflamed HaCaT cells. Through a histopathological analysis, we were able to confirm the suppressive effects of OP-D on skin thickening and the mast cell activation in AD-like mouse back skin tissues stimulated by DNCB. In addition, we detected significant decreases in cytokine expression levels through multiplex assessment assays of the OP-D-treated mice blood. We observed the anti-inflammatory effect of OP-D in the spleen, causing weight loss in the spleen and in the mRNA expression levels related to diverse cytokines. In human keratinocytes inflamed by TNF-α, OP-D inhibited p38 and ERK protein activation and showed a reduction of NF-κB nuclear translocation. Furthermore, OP-D attenuated pro-inflammatory cytokine mRNA expressions in TNF-α-inflamed HaCaT cells. Accordingly, we came to the conclusion that OP-D is a potential natural drug which can be used in order to treat inflammatory skin diseases, such as AD.


Connecting Women in Neurosciences: A networking project to sustain mutual empowerment in young, female clinical neuroscientists.
Simultaneously acquiring broad clinical knowledge and scientific expertise is a major challenge for young clinical scientists. Female researchers may face additional hurdles in their career, for example, due to unconscious bias. We aimed to address clinical, research, and gender-related challenges among young female clinical neuroscientists. We implemented a peer-led networking group dedicated to increasing clinical and scientific knowledge, improve soft skills, and encourage exchange between fellow residents. In monthly meetings, two participants hold short presentations on a clinical topic or scientific method, followed by a discussion and feedback to the presenter. Afterwards, participants network and discuss challenges they face in their daily experience. Nine neurology residents at a Swiss University Hospital with ≤3 years of training participated in the Connecting Women in Neurosciences project from August 2020 to June 2021. In a qualitative evaluation, participants reported they felt empowered by these meetings and profited from their new network. We identified several challenges in combining clinical and research activities, some of which participants perceived to be gender-related. In addition to women-only meetings, we will promote events addressing all interested researchers. Peer-to-peer networking is an easy and low-budget intervention to encourage female residents to engage in research activities, profit from each other's expertise, and promote interdisciplinary teamwork. It can provide a protected environment to discuss and overcome in particular gender-related challenges. We encourage young colleagues to regularly engage in structured networking activities with their local peers.


Protein phosphatase activity in cyanobacteria: consequences for microcystin toxicity analysis.
Hepatotoxic microcystin levels in cyanobacteria (blue-green algae) were assessed by an assay based on inhibition of protein phosphatases type 1 (PP1) and type 2A (PP2A) in crude chicken forebrain extracts using 32P-labelled glycogen phosphorylase as substrate. While cyanobacteria are reported to be devoid of phosphorylase phosphatase activity, two samples obtained from cyanobacterial scums, containing predominantly Anabaena circinalis, were found to contain high levels of a phosphorylase phosphatase activity which completely masked the presence of microcystin. Furthermore, samples containing predominantly Microcystis aeruginosa but increasing Anabaena circinalis contained sufficient phosphorylase phosphatase activity to cause a fourfold underestimation of microcystin levels. Thus, protocols for microcystin toxicity analysis should take into account the possible presence of endogenous phosphatase activity, thereby preventing underestimation of toxin levels.


Effect of maternal selenium deficiency on the teratogenicity of methylmercury.
The teratogenicity of methylmercury was examined in selenium-deficient and selenium-sufficient mice of the ICR strain. A single oral dose of 75 mumol/kg of methylmercuric chloride (MMC) on day 10, 11 or 12 of gestation caused similar incidences of a cleft palate in the selenium-deficient and selenium-sufficient mice. The treatment on day 11 led to the highest incidence of a cleft palate in both groups. These results suggest that a maternal selenium deficiency has no effect on the incidence or on the critical gestational period for an MMC-induced cleft palate.


Integrated analysis of expression profiles with meat quality traits in cattle.
MicroRNAs (miRNAs) play a vital role in improving meat quality by binding to messenger RNAs (mRNAs). We performed an integrated analysis of miRNA and mRNA expression profiling between bulls and steers based on the differences in meat quality traits. Fat and fatty acids are the major phenotypic indices of meat quality traits to estimate between-group variance. In the present study, 90 differentially expressed mRNAs (DEGs) and 18 differentially expressed miRNAs (DEMs) were identified. Eighty-three potential DEG targets and 18 DEMs were used to structure a negative interaction network, and 75 matching target genes were shown in this network. Twenty-six target genes were designated as intersection genes, screened from 18 DEMs, and overlapped with the DEGs. Seventeen of these genes enriched to 19 terms involved in lipid metabolism. Subsequently, 13 DEGs and nine DEMs were validated using quantitative real-time PCR, and seven critical genes were selected to explore the influence of fat and fatty acids through hub genes and predict functional association. A dual-luciferase reporter and Western blot assays confirmed a predicted miRNA target (bta-miR-409a and PLIN5). These findings provide substantial evidence for molecular genetic controls and interaction among genes in cattle.


A positive association between the prevalence of circadian syndrome and overactive bladder in United States adults.
To explore the association between the prevalence of circadian syndrome (CircS) and overactive bladder (OAB). Cross-section analysis was based on the National Health and Nutrition Examination Survey 2005-2018. Data regarding OAB was collected from questionnaires. The association between the prevalence of CircS and OAB was elucidated using three multivariable logistic regression models. Stratified and interaction analyses were performed to find whether some factors can modify the association. Totally 8,033 males and 8,065 females were included. People with CircS had a significantly higher prevalence of OAB compared to the non-CircS group in the fully-adjusted model (OR = 1.238, 95%CI 1.080-1.419). A significant positive correlation between the number of CircS components and the prevalence of OAB was observed when the components were ≥ 6 (OR = 1.975, 95%CI 1.463-2.665). No significant interaction was seen in the three models. There is a positive association between the prevalence of CircS and OAB. When the number of components is ≥6, the prevalence of OAB shows a strongly positive correlation with the number of CircS components.


Inherited Ventricular Arrhythmias: The Role of the Multi-Subunit Structure of the L-Type Calcium Channel Complex.
The normal heartbeat is conditioned by transient increases in the intracellular free Ca2+ concentration. Ca2+ influx in cardiomyocytes is regulated by the activity of the heteromeric L-type voltage-activated CaV1.2 channel. A complex network of interactions between the different proteins forming the ion channel supports the kinetics and the activation gating of the Ca2+ influx. Alterations in the biophysical and biochemical properties or in the biogenesis in any of these proteins can lead to serious disturbances in the cardiac rhythm. The multi-subunit nature of the channel complex is better comprehended by examining the high-resolution three-dimensional structure of the closely related CaV1.1 channel. The architectural map identifies precise interaction loci between the different subunits and paves the way for elucidating the mechanistic basis for the regulation of Ca2+ balance in cardiac myocytes under physiological and pathological conditions.


Two class II D-tagatose-bisphosphate aldolases from enteric bacteria.
Escherichia coli, Salmonella enterica, Klebsiella pneumoniaeand Klebsiella oxytocawere found to contain two D-tagatose 1,6-bisphosphate (TagBP)-specific aldolases involved in catabolism of galactitol (genes gatY gatZ) and of N-acetyl-galactosamine and D-galactosamine (genes kbaY kbaZ,also called agaY agaZ). The two aldolases were closely related (> or = 53.8% identical amino acids) and could substitute for each other in vivo. The catalytic subunits GatY or KbaY alone were sufficient to show aldolase activity. Although substantially shorter than other aldolases (285 amino acids, instead of 358 and 349 amino acids), these subunits contained most or all of the residues that have been identified as essential in substrate/product recognition and catalysis for class II aldolases. In contrast to these, both aldolases required subunits GatZ or KbaZ (420 amino acids) for full activity and for good in vivo and in vitro stability. The Z subunits alone did not show any aldolase activity. Close relatives of these new TagBP aldolases were found in several gram-negative and gram-positive bacteria, e.g., Streptomyces coelicolor.


Assessment of compression and strength of divergent screws mounted on miniplates and dynamic compression plate in fixation of mandibular body fractures: an in vitro study.
Achieving compression in the fracture line gap by open reduction and internal fixation leads to more primary bone healing and therefore hastens the healing process and patient's rehabilitation. We aimed to evaluate the application of compression by a modification in screw insertion in miniplates to improve the efficiency of the previous relevant methods. In this in vitro experimental study 20 sheep hemimandibles were prepared. Following intentional fracturing of the hemimandibular bodies, the specimens were divided into two: the control and study groups (n=10 each). The control group was fixed with straight four-hole dynamic compression plate (DCP) without a bar and with parallel screws. The study group was fixed with a straight four-hole miniplate without a bar. Screws were divergently inserted into the bone with an angle of 45°. The differences in the fracture line gap were measured before and after fixation considering the indicators of compression. The strength of the fixation was also assessed with a universal testing machine. The control group provided more compression than the study group (p=0.4). There was no difference in the strength of fixation between the two groups. It is concluded that the application of the miniplates with divergent screws instead of DCP could encompass the advantages of both perspectives such as intraoral incisions and compressive force and prevent the disadvantages of compression plates such as hard adaptation.


Cell cycle events during the development of the silk glands in the mulberry silkworm Bombyx mori.
Silk glands of the mulberry silkworm Bombyx mori are long and paired structures originating from the labial region and are anatomically and physiologically divided into three major compartments, the anterior, middle and posterior silk glands. The silk gland morphogenesis is complete by 8 days post egg laying. Extensive growth of silk glands during the larval stages is due to increase in tissue mass and not cell number. The cells in a completely formed silk gland pursue an endoreplicative cell cycle, and the genome undergoes multiple rounds of replication without mitosis or nuclear division. The expression patterns of cyclin B (mitotic cyclin) and cyclin E (G1 cyclin, essential for G1/S transition in both mitotic and endoreplicative cell cycles) in the course of silk gland development revealed that mitotic cell divisions take place only in the apex of the growing silk gland. However, the persistence of another mitotic focus in the middle silk gland even when the growing apex has moved well past this zone suggested the continued operation of mitosis for a while in this restricted region. The lack of cyclin B expression and abundance of cyclin E in the rest of the areas confirmed an alternation of the G1 and S phases of the cell cycle without an intervening mitotic phase. No expression of cyclin B was noticed anywhere in the silk glands after stage 25 of embryogenesis, indicating a complete switch over to the endomitotic mode of the cell cycle. The onset of expression of various genes encoding different silk proteins correlated with the onset of endomitotic events.


[Computerized tomography, magnetic resonance, and nuclear medicine in the non-invasive diagnosis of focal nodular hyperplasia of the liver].
Focal nodular hyperplasia (FNH) is a benign liver lesion requiring a prompt diagnosis and a conservative management. Aim of our study was to prospectively integrate enhanced CT, MRI, nuclear medicine in the noninvasive diagnosis of FNH. 20 FNH lesions (diameter ranging 1.5-13 cm) in 18 asymptomatic patients were investigated with MRI and nuclear medicine. MRI examinations were performed with a 1.5 T superconducting system (Philiphs NT) by acquiring T1-weighted, T2-weighted, T2-weighted fat-suppressed Turbo Spin-Echo and dynamic MRI sequences, with breath-hold T1-weighted Turbo Field Echo and Gd-DTPA. Nuclear medicine included in all cases 99mTc sulfur colloid or 99mTc iminodiacetic acid studies. Diagnostic sensitivity of MRI was compared with that of nuclear medicine, and the sensitivity of the two modalities combined. The definitive diagnosis was made by percutaneous core-needle biopsy (12 lesions), surgery (2 lesions) and longterm follow-up (6 lesions). In 13 lesions larger than 3.5 cm typical findings, such as the central scar and homogeneous pattern, were observed in 9 cases (75%) with enhanced CT, in 10 cases (83%) with unenhanced MRI and in 11 cases (91%) after gadolinium injection. Hepatobiliary scintigraphy showed increased tracer uptake in delayed scans in 10/12 cases (83%) while sulfur colloid studies were diagnostic in 2/12 cases only (16%), showing the same tracer uptake than the surrounding liver parenchyma. In 7 lesions smaller than 3.5 cm, only 1 case showed typical findings; dynamic MRI showed typical early hypervascularity in 5 of the 6 remaining lesions (71%) which persisted on late images. In lesions smaller than 3.5 cm, sulfur colloid studies were diagnostic in 1/7 lesions (14%) and hepatobiliary scintigraphy in 3/7 lesions (42%). Considering all 20 lesions, MRI-nuclear medicine integration performed better than MRI alone. Enhanced MRI and biliary scintigraphy are important tools to make the final diagnosis of FNH. In small lesions both examinations should performed, but in large lesions MRI may be sufficient to make an unquestionable diagnosis, limiting hepatobiliary scintigraphy to questionable cases.


Pathogenic roles of altered calcium channels and transporters in colon tumorogenesis.
Cytosolic calcium [Ca2+]cyt signaling plays a critical role in the regulation of multiple cellular functions, and Ca2+ channels/transporters are important to regulate calcium homeostasis whose abnormality may contribute human tumorogenesis including colorectal cancer (CRC). In this review, we summarized and discussed the current knowledge on pathogenic roles of the altered [Ca2+]cyt and Ca2+ channels/transporters like SOCE, TRP channels, SERCA and Na+/Ca2+ exchangers in CRC tumorigenesis and progression. Understanding the detailed molecular mechanisms underlying the effects of [Ca2+]cyt on CRC is essential to develop Ca2+ channels/transporters as diagnostic and therapeutic targets. Targeting Ca2+ signaling for cancer therapy has become an emerging research area nowadays, although our knowledge about the roles of Ca2+ channels/transporters in tumorigenesis is still in the early stage, we still believe that they will act as novel preventive/therapeutic targets for CRC with potentially extensive clinical significance.


Preparation and characterization of cellulose nanocrystals from spent edible fungus substrate.
Spent edible fungus substrates were identified as potential sources to produce cellulose derivatives, namely purified cellulose and dicarboxyl cellulose nanocrystal (DCNC). Purified celluloses were obtained via chemical treatments and then oxidized by sequential periodate-chlorite without mechanical process. The structural properties of the DCNCs were characterized by transmission electron microscopy (TEM), Fourier-transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD) and thermal gravimetric analysis (TGA). XRD results showed that the cellulose I structure was maintained, however, the crystallinity index decreased after oxidation process. The initial pyrolysis temperature of DCNCs ranged from 242 to 344 °C. TEM results revealed that DCNC was rod-shaped with an average length and width of 130.88 nm and 7.3 nm, respectively. The average specific surface area (SSA) was 366.67 m2 g-1 . The carboxyl content was around 3.485 mmol g-1 . Finally, the adsorption capacity for contaminations was 76.98, 126.22, 64.44 and 9.63 mg g-1 for copper ion (Cu2+ ), lead ion (Pb2+ ), chromium (Cr3+ ) and amoxicillin (AMX), respectively. This work showed a sequentially chemical oxidation for preparing nanocellulose from secondary agricultural waste with many functional applications. © 2021 Society of Chemical Industry.


Blood pressure, metabolic and autonomic responses to insulin and intralipid® infusion in chagasic patients.
Intralipid(®) and heparin infusion results in increased blood pressure and autonomic abnormalities in normal and hypertensive individuals. To evaluate insulin sensitivity and the impact of Intralipid(®) and heparin (ILH) infusion on hemodynamic, metabolic, and autonomic response in patients with the indeterminate form of Chagas' disease. Twelve patients with the indeterminate form of Chagas' disease and 12 healthy volunteers were evaluated. Baseline blood pressure and heart rate were similar in both groups. Plasma noradrenaline levels were slightly increased in the Chagas' group. After insulin tolerance testing (ITT), a significant decline was noted in glucose in both groups. ILH infusion resulted in increased blood pressure in both groups, but there was no significant change in plasma noradrenaline. The low-frequency component (LF) was similar and similarly increased in both groups. The high-frequency component (HF) was lower in the Chagas' group. Patients with the indeterminate form of Chagas' disease had increased sympathetic activity at baseline and impaired response to insulin. They also had a lower high-frequency component and impaired baroreflex sensitivity at baseline and during Intralipid(®) and heparin infusion.


Predisposing conditions and risk factors for development of symptomatic meningioma in adults.
Risk factors and predisposing factors for the development of symptomatic meningioma during adult life are not well known. Data from 306 consecutive patients with primary meningioma were collected retrospectively in a hypothesis-generating study. Factors studied included localisation of tumours, blood group typing, and risk factors, such as diabetes mellitus, coronary arterial disease, hypertension, rheumatoid arthritis, bronchial asthma, smoking, obesity, and second primary tumour. Case-control analysis of putative risk factors was carried out using a control data set from the German East-West Health Survey (n=7466, age range 25-69 years). Patients and controls were matched for age, gender, geographic area, and time of data collection. Rh(D) positive cases were significantly less frequent in the patient group compared to controls (p=0.01). Pre-existing diabetes was associated with meningioma in middle-aged (40-69 years) patients (odds ratio, OR 13.94-4.30, p=0.001-0.05). In female patients, arterial hypertension was significantly associated with occurrence of meningioma in the age group 60-69 years (OR=2.23, p=0.041). Rheumatoid arthritis had a negative association with meningioma in both males and females in the age groups above 50 years (OR 0.19-0.27, p=0.02-0.034). Bronchial asthma, smoking, and obesity were not significantly associated with meningioma. A second primary tumour was present in 12 cases. The most frequent combination was meningioma and breast cancer (5/12). This study shows statistically significant association of some co-morbidities with symptomatic meningioma in adults. Areas of interest have been identified where further research would be necessary.


Highly efficient gene knockout system in the maize pathogen Colletotrichum graminicola using Agrobacterium tumefaciens-mediated transformation (ATMT).
Colletotrichum graminicola, a hemibiotrophic pathogenic fungus, is the causal agent of anthracnose of maize, which causes significant yield losses worldwide, especially in warm and humid maize production regions. An efficient targeted genes knockout protocol is crucial to explore molecular mechanisms of fungal virulence to the host. In this study, we established a gene knockout transformation system by employing Agrobacterium tumefaciens-mediated transformation to knockout genes in M 1.001 strain of C. graminicola. The conidia germination status, induction medium type, and ratio of Agrobacterium cell and conidia suspension were optimized for the knockout of CgBRN1(OR352905), a gene relating to the fungal melanin biosynthesis pathway. Additionally, CgPKS18 (OR352906) and CgCDC25 (OR352903) were knocked out to test the applicability of the gene knockout transformation system. In this established system, transformation efficiency was 176 transformants per 1 × 105 conidia and the homologous recombination efficiency was 53.3 to 75%. Furthermore, disease index, lesion number and lesion size caused by the three above-mentioned mutant strains were found to be reduced significantly compared to the wild-type strain, which indicated reduction in fungal virulence due to the lack of those genes.


Molecular cloning and characterization of an enzyme hydrolyzing p-nitrophenyl alpha-D-glucoside from Bacillus stearothermophilus SA0301.
Bacillus stearothermophilus SA0301 produces an extracellular oligo-1,6-glucosidase (bsO16G) that also hydrolyzes p-nitrophenyl alpha-D-glucoside (Tonozuka et al., J. Appl. Glycosci., 45, 397-400 (1998)). We cloned a gene for an enzyme hydrolyzing p-nitrophenyl alpha-D-glucoside, which was different from the one mentioned above, from B. stearothermophilus SA0301. The k(0)/K(m) values of bsO16G for isomaltotriose and isomaltose were 13.2 and 1.39 s(-1).mM(-1) respectively, while the newly cloned enzyme did not hydrolyze isomaltotriose, and the k(0)/K(m) value for isomaltose was 0.81 s(-1).mM(-1). The primary structure of the cloned enzyme more closely resembled those of trehalose-6-phosphate hydrolases than those of oligo-1,6-glucosidases, and the cloned enzyme hydrolyzed trehalose 6-phosphate. An open reading frame encoding a protein homologous to the trehalose-specific IIBC component of the phopshotransferase system was also found upstream of the gene for this enzyme.


6-Mercaptopurine pharmacokinetics after use of azathioprine in renal transplant recipients with intermediate or high thiopurine methyl transferase activity phenotype.
Prevention of allograft transplant rejection by the immunosuppressive 6-thiopurine drug azathioprine is limited by haematological toxicity (leucopenia or agranulocytosis). This toxicity is particularly apparent in subjects with low thiopurine methyltransferase activity (TPMTase) phenotype (1% in the Caucasian population). The thiopurine derivative 6-mercaptopurine is the active metabolite of azathioprine, and it would be of interest to measure, after validation of plasma measurements, the mean values of the pharmacokinetic parameters in transplant patients with high or intermediate TPMTase phenotypes (85 and 14% of the Caucasian population, respectively). We measured erythrocyte TPMTase activity in 103 kidney transplant recipients of high or intermediate phenotype and calculated, after a test dose of azathioprine, the mean values of the pharmacokinetic parameters for 6-mercaptopurine. We also compared these values with the same parameters from one subject with low TPMTase activity phenotype. The mean observed area under the plasma concentration-time curve (AUC) was 190+/-140 ng mL(-1) h and the elimination rate constant (Kel) was 1.92+/-1. The pharmacokinetic parameters (AUC, Kel, t1/2el (the elimination half-life)) of 6-mercaptopurine in transplant patients are normally distributed and suitable for acceptance as a gold standard value for this population of Caucasian transplant patients. It seems useful to calculate these parameters, representative of the systemic exposure of individual patients to the drug, before prescribing these subjects azathioprine immunosuppressive treatment. In subjects with low TPMTase phenotype these pharmacokinetic measurements could also be an index of dose reduction.


Red blood cell enzyme abnormalities in patients treated with chemotherapy.
Sxiteen red blood cell enzyme activities and fetal haemoglobin level have been assayed in 60 patients treated for haematologic or nonhaematologic malignant diseases with various combinations of cytostatic drugs. Acquired enzyme deficiency was found in 20 patients. The most frequently decreased activities were those of G6PD (12 cases), PK (seven cases), PFK (six cases) and AK (three cases). In many patients decreased activity of some enzymes contrasted with increased activity of others such as ALD, TPI, G3PD, PGK, ENOL and 6PGD. The number of abnormalities seems to be related to the duration of the treatment. Incidence of enzyme deficiencies was similar in patients previously treated or not with radiotherapy. Enzyme abnormalities were correlated neither with haemoglobin level nor with reticulocyte count. They were very similar to those observed in primary acquired dyserythropoietic and preleukaemic states. Their prognostic value and mechanism are discussed.


Congenital malformations of the human brainstem.
Congenital disorders of the brainstem often result in multiple severe neurodevelopmental problems. With the advent of magnetic resonance imaging and discovery of genes directing brainstem formation, a more coherent clinical picture of these disorders is emerging. Proper evaluation, management, and counseling for these disorders rests on the clinician having a framework through which to approach them.


[Association on Sturge Weber and Klippel Trenaunay Weber syndromes. Apropos of 2 cases].
The present paper describes two patients with Sturge-Weber and Klippel Trénaunay-Weber syndromes. Some etiopathogenic factors are analyzed. We suggest that the association of both diseases in the same patient may be due to a single autosomal dominant gene.


Stereotactic body radiation therapy in centrally and superiorly located stage I or isolated recurrent non-small-cell lung cancer.
To evaluate the efficacy and adverse effects of image-guided stereotactic body radiation therapy (SBRT) in centrally/superiorly located non-small-cell lung cancer (NSCLC). We delivered SBRT to 27 patients, 13 with Stage I and 14 with isolated recurrent NSCLC. A central/superior location was defined as being within 2 cm of the bronchial tree, major vessels, esophagus, heart, trachea, pericardium, brachial plexus, or vertebral body, but 1 cm away from the spinal canal. All patients underwent four-dimensional computed tomography-based planning, and daily computed tomography-on-rail guided SBRT. The prescribed dose of 40 Gy (n = 7) to the planning target volume was escalated to 50 Gy (n = 20) in 4 consecutive days. With a median follow-up of 17 months (range, 6-40 months), the crude local control at the treated site was 100% using 50 Gy. However, 3 of 7 patients had local recurrences when treated using 40 Gy. Of the patients with Stage I disease, 1 (7.7%) and 2 (15.4%) developed mediastinal lymph node metastasis and distant metastases, respectively. Of the patients with recurrent disease, 3 (21.4%) and 5 (35.7%) developed mediastinal lymph node metastasis and distant metastasis, respectively. Four patients (28.6%) with recurrent disease but none with Stage I disease developed Grade 2 pneumonitis. Three patients (11.1%) developed Grade 2-3 dermatitis and chest wall pain. One patient developed brachial plexus neuropathy. No esophagitis was noted in any patient. Image-guided SBRT using 50 Gy delivered in four fractions is feasible and resulted in excellent local control.


Structural brain imaging as a diagnostic tool in dementia, why and how?
The demands for more people to be investigated due to cognitive failure and suspected dementia are increasing as increasing numbers of us get older and the incidence of dementia increases. An important part of a dementia study includes the structural imaging of the brain. Two imaging techniques, Computed Tomography (CT) and Magnetic Resonance Imaging (MRI), are used in this context. They differ in many ways and one question is which of the methods should be used in the first instance. Considering the large number of investigations to be expected in the future it is vital that they be cost-effective. Structural imaging aims partly to find secondary causes of cognitive failure and partly to provide support in the differential diagnostic reasoning. The methods differ; CT is significantly cheaper but exposes the patient to radiation, MRI is expensive but does not use X-rays. MRI provides better imaging of cerebrovascular lesions than CT as well as better imaging of structures near the skull base. The difference in diagnostic accuracy is small and it is doubtful whether that difference justifies the large difference in cost.


Analysis of the interaction between A1 R and A2A R proteins in living cells based on FRET imaging and batch processing method.
The quantitative FRET analysis of living cells is a tedious and time-consuming task for freshman lacks technical training. In this study, FRET imaging and batch processing method were combined to analyze reagents-induced interactions of A1 R and A2A R on cell membranes. Results showed that the method had taken less time than if cell-by-cell was analyzed. The accuracy and repeatability of FRET efficiency values were likewise improved by removing the interference from anthropogenic factors. Then this method was applied to rapidly analyze acetaldehyde-induced interactions, which analyzed hundreds of single-cell trends by one operation, and the results revealed that interactions were consistently attenuated in LX-2 cells, and statistical differences appeared after 30 min. Combined with batch processing method, procedures of cells FRET analysis have been greatly simplified without additional technical work, which has broad prospects in large-scale analysis of cellar protein interaction.


Immunogenicity of Hepatitis B Vaccine in HIV Exposed Uninfected Infants.
There is paucity of knowledge about the immunogenicity of vaccines in infants who have been exposed to HIV in-utero but have remained uninfected. The authors studied the immunogenicity of 3 doses of recombinant hepatitis B vaccine at 6,10,14 wk of age in HIV exposed but uninfected (HEU) infants. After 3 mo of last dose of the vaccine, out of 26 infants, 23 (89.5 %) infants were found to be responders (Anti HBs IgG titres ≥ 10 mIU/ml) and 3 (11.5 %) babies remained non responders (Anti HBs IgG titres < 10 mIU/ml). The proportion of babies who were non responders were higher when compared to similar studies done on unexposed and uninfected infants, suggesting a poorer immunogenicity of hepatitis B vaccine in these infants.


Effect of Gibberellin Biosynthesis Inhibitors on Native Gibberellin Content, Growth and Floral Initiation in Sorghum bicolor.
CCC, uniconazol, ancymidol, prohexadione-calcium (BX-112), and CGA 163'935, which represent three groups of gibberellin (GA) biosynthesis inhibitors, were applied as a soil drench to Sorghum bicolor cultivars 58M (phyB-1, phytochrome B-deficient mutant) and 90M (phyB-2, equivalent phenotypically to wild type, PHYB, except for small differences in flowering dates). The inhibitors that block steps before GA12 (CCC, uniconazol, and ancymidol) lowered the concentrations of all endogenous early-C13alpha-hydroxylation pathway GAs found in sorghum: GA12, GA53, GA44, GA19, GA20, GA1, and GA8. In contrast, the inhibitors that block the conversion of GA20 --> GA1, (CGA 163'935 and BX-112) drastically reduced GA1 and GA8 levels, but they either did not change or caused accumulation of intermediates from GA12 to GA20. Combinations of pre-GA12 inhibitors and GA3 plus GA1 strongly reduced GAs other than GA1 and GA3. Each of these compounds inhibited shoot growth in both cultivars and delayed floral initiation in 90M. Floral initiation of 58M was also delayed by CCC, uniconazol, and ancymidol but not by CGA 163'935 and BX-112. This separation of shoot elongation from floral initiation in sorghum is novel. Both inhibition of shoot growth and delayed floral initiation were almost completely relieved by a mixture of GA3 and GA1 in both 58M and 90M. This observation, plus the much lower levels of endogenous GA3 than of GA1 observed in these experiments, implies that GA1 is the major endogenous GA active in shoot elongation. CGA 163'935 and BX-112 also failed to promote tillering in 58M, whereas inhibitors active before GA12 did so. The possibility that the GA20 --> GA1 inhibitors fail to block flowering and promote tillering in 58M because biosynthetic intermediates between GA12 and GA20 accumulate and/or because 58M is altered in GA metabolism in this same region of the biosynthetic pathway is discussed. Key Words. Gibberellin biosynthesis inhibitors-Phytochrome B-Flowering-Sorghum


Distraction histogenesis of an anastomosed artery.
This study examined the possibility of an adaptive reaction of anastomosed arteries under tension during a distraction lengthening procedure in the tibiae of rabbits. After an osteotomy at the mid tibia, the posterior tibial arteries were transected and anastomosed. Using a pair of small external fixators, the tibiae were distracted at a rate of 0.5mm/day (groups I-IV rabbits). Three weeks after 25% lengthening, the patency and histology of the arteries were examined. Angiography revealed that all of the anastomosed arteries were patent, and intimal hyperplasia was a constant finding. The mean thickness of the intima of the lengthened segment in group I was 60.4 microm, which is 5.0, 3.4 and 2.1 times higher that of the controls in groups IV (un-manipulated arteries, 12.2 microm), III (unlengthened but anastomosed arteries, 17.8 microm) and II (lengthened but untouched arteries, 28.7 microm), respectively. These results show that an anastomosed artery can maintain its patency at a certain level and speed of distraction lengthening. Therefore, it is possible that distraction lengthening and vascular anastomoses can be performed simultaneously provided there is careful monitoring of the circulation.


Clinical manifestations of primary hyperparathyroidism before and after parathyroidectomy. A case-control study.
There has been an National Institutes of Health consensus meeting concerning the management of patients with "asymptomatic" primary hyperparathyroidism, yet there is no clear definition of this condition. The authors, therefore, documented the clinical manifestations and frequencies of these manifestations in unselected patients with primary hyperparathyroidism and determined whether these clinical manifestations resolved after parathyroidectomy. The authors studied 152 unselected consecutive patients with primary hyperparathyroidism and 132 control patients with nontoxic thyroid disorders who were treated by parathyroidectomy or thyroidectomy, respectively, between January 1986 and June 1991. All patients received a questionnaire during their initial office visits and the same questionnaire again after their operations. Patients were also questioned about their perception of the success of the operation. Eighty percent of the parathyroid patients and 70.5% of the thyroid patients completed the questionnaires, and the mean follow-up time was 20 months. Only 7 (4.6%) patients with primary hyperparathyroidism had no symptoms, and 26 (17.1%) had no associated conditions despite 74.3% of these patients having serum calcium levels less than 12 mg/dL. Symptoms including fatigue, exhaustion, weakness, polydipsia, polyuria, nocturia, joint pain, bone pain, constipation, depression, anorexia, nausea, heartburn, and associated conditions, including nephrolithiasis, and hematuria occurred more often in patients with primary hyperparathyroidism than in the thyroid control patients (p < 0.05). After parathyroidectomy, only eight (5.3%) patients failed to have any improvement in symptoms or associated conditions. Fifty-seven percent of the parathyroid patients versus 30% of the thyroid patients felt better overall after the operation, strength subjectively improved in 29% of parathyroid patients versus 13% in thyroid patients; thirty-seven percent of the parathyroid patients versus 13% of the thyroid patients claimed they were less depressed. When examined thoroughly, few patients with primary hyperparathyroidism were asymptomatic or without associated conditions, and most patients experienced subjective improvement after successful parathyroidectomy.


Influence of a dorsomedial hypothalamus lesion on the circadian changes in the enzyme development and activity of intestinal brushborder membranes in the rat.
Circadian changes in activities and electrophoretic pattern of the intestinal brushborder enzymes alkaline phosphatase (EC 220.127.116.11), isomaltase (EC 188.8.131.52) and sucrase (EC 184.108.40.206) of normal rats and of rats with a lesion of the dorsomedial hypothalamus (DMH-rats) were studied. In contrast to the rhythm found in normal rats, there are no significant differences between the average enzyme activities in the light or dark period for the DMH-rats. The peak ratio in the electrophoretic pattern for sucrase-isomaltase (SI) over pro-sucrase-isomaltase (pro-SI) changes over 24 hr with higher values during the active (dark) period of the normal rat. In DMH-rats this ratio is much higher, but no significant difference could be demonstrated between the values near the acrophase in these rats.


Differential dependence of hypoxia-inducible factors 1 alpha and 2 alpha on mTORC1 and mTORC2.
Constitutive expression of hypoxia-inducible factor (HIF) has been implicated in several proliferative disorders. Constitutive expression of HIF1 alpha and HIF2 alpha has been linked to a number of human cancers, especially renal cell carcinoma (RCC), in which HIF2 alpha expression is the more important contributor. Expression of HIF1 alpha is dependent on the mammalian target of rapamycin (mTOR) and is sensitive to rapamycin. In contrast, there have been no reports linking HIF2 alpha expression with mTOR. mTOR exists in two complexes, mTORC1 and mTORC2, which are differentially sensitive to rapamycin. We report here that although there are clear differences in the sensitivity of HIF1 alpha and HIF2 alpha to rapamycin, both HIF1 alpha and HIF2 alpha expression is dependent on mTOR. HIF1 alpha expression was dependent on both Raptor (a constituent of mTORC1) and Rictor (a constitutive of mTORC2). In contrast, HIF2 alpha was dependent only on the mTORC2 constituent Rictor. These data indicate that although HIF1 alpha is dependent on both mTORC1 and mTORC2, HIF2 alpha is dependent only on mTORC2. We also examined the dependence of HIF alpha expression on the mTORC2 substrate Akt, which exists as three different isoforms, Akt1, Akt2, and Akt3. Interestingly, the expression of HIF2 alpha was dependent on Akt2, whereas that of HIF1 alpha was dependent on Akt3. Because HIF2 alpha is apparently more critical in RCC, this study underscores the importance of targeting mTORC2 and perhaps Akt2 signaling in RCC and other proliferative disorders in which HIF2 alpha has been implicated.


Comparison of the Canal Master endodontic instrument and K-type files for enlargement of curved root canals.
Twenty mandibular molars with 40 mesial curved root canals were instrumented with a circumferential technique using K files and the Canal Master instrumentation technique by junior dental students whose only endodontic experience had been completion of a preclinical endodontic course. Preoperative and postoperative radiographs were taken facially and mesially with instruments in place. The canals were evaluated radiographically for transportation 1 mm and 4 mm from the apex. The mesial roots were cross-sectioned at the same level. The sections were evaluated for roundness of the canal preparation. It was concluded that the Canal Master instrumentation technique transported the root canal less and produced rounder preparations than the K file circumferential technique. There appears to be a greater tendency for breakage with the Canal Master.


Medication adherence and rate of nicotine metabolism are associated with response to treatment with varenicline among smokers with HIV.
PLWHA who smoke have shown lower cessation rates within placebo-controlled randomized trials of varenicline. Adherence and rate of nicotine metabolism may be associated with quit rates in such clinical trials. This secondary analysis of a randomized placebo-controlled trial of varenicline for smoking among PLWHA (N = 179) examined the relationship between varenicline adherence (pill count, ≥80% of pills), nicotine metabolism (based on the nicotine metabolite ratio; NMR) and end-of-treatment smoking cessation (self-reported 7-day point prevalence abstinence, confirmed with carbon monoxide of ≤ 8 ppm, at the end of treatment; EOT). Combining varenicline and placebo arms, greater adherence (OR = 1.011, 95% CI:1.00-1.02, p = 0.051) and faster nicotine metabolism (OR = 3.08, 95% CI:1.01-9.37, p = 0.047) were related to higher quit rates. In separate models, adherence (OR = 1.009, 95% CI:1.004-1.01, p < 0.001) and nicotine metabolism rate (OR = 2.04, 95% CI:1.19-3.49, p = 0.009) interacted with treatment arm to effect quit rates. The quit rate for varenicline vs. placebo was higher for both non-adherent (19% vs. 5%; χ2[1] = 2.80, p = 0.09) and adherent (35% vs. 15%; χ2[1] = 6.51, p = 0.01) participants, but the difference between treatment arms was statistically significant only for adherent participants. Likewise, among slow metabolizers (NMR < 0.31), the varenicline quit rate was not significantly higher vs. placebo (14% vs. 5%; χ2[1] = 1.17, p = 0.28) but, among fast metabolizers (NMR ≥ 0.31), the quit rate for varenicline was significantly higher vs. placebo (33% vs. 14%; χ2[1] = 4.43, p = 0.04). Increasing varenicline adherence and ensuring that fast nicotine metabolizers receive varenicline may increase quit rates for PLWHA.


Attenuation of dermal wounds via downregulating oxidative stress and inflammatory markers by protocatechuic acid rich n-butanol fraction of Trianthema portulacastrum Linn. in wistar albino rats.
Oxidative stress and inflammation contribute as a key factor for retarding the process of dermal wound healing. Trianthema portulcastrum Linn. (TP) leaves reported to possess antioxidant, antifungal, anti-inflammatory and antibacterial properties, which could make TP a promising wound healing agent. The current study was aimed to estimate the antioxidant potential of the fractionated hydroethanolic extract of TP leaves and evaluate wound healing activity by excision and incision wound models along with the assessment of possible underlying mechanism. Ethyl acetate, chloroform and n-butanol fractions of the hydroethanolic extract of TP leaves were examined for in vitro antioxidant ability by DPPH method. Strongest antioxidant activity bearing n-butanol fraction (nBuTP) was further analyzed quantitatively by High Performance Liquid Chromatography coupled with Diode Array Detector (HPLC-DAD). Wound healing potential of nBUTP using excision and incision wound model was studied. Wistar albino rats were randomly divided into four groups, containing six animals in each group; group I served as control treated with simple ointment base, group II was standard group, treated with povidone-iodine ointment USP (5%), group III treated with nBuTP 5% w/w ointment, and group IV treated with nBuTP 10%w/w ointment. All the groups were topically applied their respective ointments, once daily, till the complete healing achieved. Wound healing was assessed by analyzing % wound closure, hydroxyproline content, epithelialization period, tensile strength, enzymatic antioxidative status and inflammatory markers. Total phenolic and flavonoid content of the extract was estimated to be 112.32±1.12 and 84.42±0.47mg/g, respectively. HPLC-DAD of nBuTP confirmed the presence of chlorogenic acid (20.74±0.03), protocatechuic acid (34.45±0.02mg/g), caffeic acid (4.31±0.03mg/g) and ferulic acid (1.43±0.01mg/g). 5% and 10%w/w nBuTP ointment significantly accelerated the wound healing process dose-dependently in both wound models, evidenced by the faster rate of wound contraction, epithelialization, increased hydroxyproline content, high tensile strength, increased antioxidant enzyme activity, decreased the level of inflammatory markers compared to the control group. Histopathological studies also revealed the dose-dependant amelioration of wound healing by re-epithelialization, collagenation and vascularization of wounded skin sample in nBuTP treated groups. These results implicate potential medicinal value of nBuTP to heal dermal wounds.


Absolute configuration and predominant conformations of a chiral crown ether-based colorimetric sensor: a vibrational circular dichroism spectroscopy and DFT study of chiral recognition.
In the present work, we report a comprehensive vibrational circular dichroism (VCD) spectroscopic study of a chiral crown ether which features an axial chiral 3.3'-diphenyl-1,1'-binaphthyl group as chiral moiety. By comparing the experimental and calculated VCD spectra, we show that the presumably very flexible crown ether preferably adopts only one ring conformation. Conformational flexibility is observed in the 2,4-dinitrophenyl-diazophenol group, which was previously introduced for colorimetric detection of primary amines and amino alcohols (Cho et al., Chirality 2011;23:349-353). The VCD spectra of the host-guest complexes with phenyl glycinol (PG) and phenyl alaninol have been studied as well. Based on the spectra calculated, it is shown that the diastereomeric complexes in general can be differentiated using VCD spectroscopy. Furthermore, the experimental VCD spectra of the complexes of the host molecule with PG support the above finding.


Acquired Morgagni hernia following coronary artery bypass graft (CABG) with successful robotic repair of hernia.
Morgagni hernia is an uncommon type of diaphragmatic hernia and commonly presents as a congenital disease. Acquired Morgagni hernias following open cardiac surgery are exceedingly rare and only reported in the pediatric population. The patient is a 70-year-old female who presented with complaints of shortness of breath and cough one year following a coronary artery bypass graft (CABG). A chest CT scan showed a large Morgagni type diaphragmatic hernia with herniated transverse colon occupying the anterior mediastinum as well as the right hemi-thorax. This hernia was successfully repaired using transabdominal robotic approach with complete resolution of patient's symptoms. This is the first reported case of acquired Morgagni type diaphragmatic hernia in an adult following open cardiac surgery. The potential etiologies for this hernia include distal extension of the median sternotomy and involvement of the anterior diaphragm, iatrogenic injury to the attenuated anterior diaphragm during pericardial window creation, or pericardial drain placement. Operative repair is the mainstay of treatment and is usually performed with a transabdominal approach since it is thought to be less challenging and allows for evaluation of the entire abdominal cavity. If primary repair cannot be achieved, then synthetic mesh may be needed to obtain a tension free and durable repair. We present a case of acquired Morgagni type diaphragmatic hernia in an adult following open cardiac surgery that was successfully repaired using a transabdominal robotic approach.


Relation disentanglement, the potential risk assessment, and source identification of heavy metals in the sediment of the Changzhao Reservoir, Zhejiang Province.
Heavy metal contamination in the water body is a distinctly important issue for the water security of the reservoir. 114 sediment samples of Changzhao Reservoir were collected to investigate the spatial (horizontal and vertical) distribution characteristics, risk assessment, and source identification of heavy metals. The concentrations of heavy metals at the surface layer of sediment were slightly higher compared with that at the middle and bottom layer sediment in the most sampling sites. The concentration of Zn and Cd was significantly different in the different depths of sediment (P ≤ 0.01, Tukey HSD test). pH and Cd were identified as the key factors for TOC in the sediment by the Boruta algorithm. The proportion of "uncontaminated to moderately contaminated" for Cd, Zn, and As in the surface layer was 84.21%, 47.37%, and 34.21%, which indicated that the quality of sediment was mostly impacted by Cd, Zn, and As. The agricultural non-point source pollution is dominant according to the source identification method of APCS-MLR. Overall, this paper presents the distribution and conversion trends of heavy metals and provides the insights of the reservoir protection in the future work.


[Food status peculiarities, anthropometric, clinica and biochemical indices at professional sportsmen].
Under steady state conditions in 66 athletes involved in weightlifting, bodybuilding, judo and taekwondo have studied features of the metabolic status. Data on matter-of-fact nutrition, body weight content within the inter-competition period, energy exchange, clinical and biochemical indices and physical acceptability indices were analyzed. As a result, the decrease indexes of metabolism at all the sportsmen and high-level caloric value at sportsmen who are engaged in weightlifting, which corresponds their energy expenditures, was revealed.


Modern Perspective on Metabolic Reprogramming in Malignant Neoplasms.
Metabolic reprogramming is one of the central features of transformed cells. Elucidation of interactions between oncogenic signaling and cell metabolic processes has become the basis for extensive studies of metabolism reprogramming in tumor tissue. The review summarizes the key results of studies on the catabolic and anabolic rearrangements in tumor cells with special emphasis on carbohydrate, lipid, amino acid, and acetate metabolism determining the cancer phenotype of cells.


Utility of 70-kV single-energy CT in depicting the extent of breast cancer for preoperative planning.
To evaluate the detectability of breast cancer and visibility of the tumor extent using 70-kV single-energy contrast-enhanced (CE) breast computed tomography (70-kV CECT) compared with CE breast magnetic resonance imaging (CEMR). Between 2013 and 2015, 110 patients with 112 breast cancer lesions who underwent breast surgery after undergoing both 70-kV CECT and CEMR were enrolled. The major axis lengths of the breast lesion were measured and compared with the pathologically determined major axes. Agreement in the measured major axes was evaluated using the intra-class correlation coefficient (ICC). Both 70-kV CECT and CEMR depicted all breast cancer lesions. The mean major axis was 3.0 (95% confidence interval [CI], 2.5-3.4) cm on CECT and 2.9 (2.6-3.3) cm on CEMR. The mean differences between the pathologically and radiologically measured major axes on 70-kV CECT and CEMR were 0.9 (0.7-1.1) and 1.0 (0.8-1.2) cm, respectively. The accuracy of the radiological major axes compared with the pathological major axes was 82.1% and 80.4% on CECT and CEMR, respectively (p = 0.81). The major axes on the two modalities demonstrated moderate agreement (ICC = 0.69, 95% CI 0.58-0.77). Pathologically and radiologically measured major axes on 70-kV CECT and CEMR demonstrated excellent agreement (ICC = 0.91, 95% CI 0.93-0.96). Low-tube voltage (70-kV) CECT is the preferred modality to identify breast cancer lesions and tumor extent for preoperative planning because it has a similar diagnostic ability to CEMR and can be performed in the supine position.


Ultrasensitive electrochemical cocaine biosensor based on reversible DNA nanostructure.
We proposed an ultrasensitive electrochemical cocaine biosensor based on the three-dimensional (3D) DNA structure conversion of nanostructure from Triangular Pyramid Frustum (TPFDNA) to Equilateral Triangle (ETDNA). The presence of cocaine triggered the aptamer-composed DNA nanostructure change from "Close" to "Open", leading to obvious faradaic impedance changes. The unique properties with excellent stability and specific rigid structure of the 3D DNA nanostructure made the biosensing functions stable, sensitive, and regenerable. The Faradaic impedance responses were linearly related to cocaine concentration between 1.0 nM and 2.0 μM with a correlation coefficient of 0.993. The limit of detection was calculated to be 0.21 nM following IUPAC recommendations (3Sb/b). It is expected that the distinctive features of DNA nanostructure would make it potentially advantageous for a broad range of biosensing, bionanoelectronics, and therapeutic applications.


Cytotoxicity of topical antimicrobial agents used in burn wounds in Australasia.
Burn sepsis is a leading cause of mortality and morbidity in patients with major burns. The use of topical antimicrobial agents has helped improve the survival of these patients. Silvazine (Sigma Pharmaceuticals, Melbourne, Australia) (1% silver sulphadiazine and 0.2% chlorhexidine digluconate) is used exclusively in Australasia, and there is no published study on its cytotoxicity. This study compared the relative cytotoxicity of Silvazine with 1% silver sulphadiazine (Flamazine (Smith & Nephew Healthcare, Hull, UK)) and a silver-based dressing (Acticoat (Smith & Nephew Healthcare, Hull, UK)). Dressings were applied to the centre of culture plates that were then seeded with keratinocytes at an estimated 25% confluence. The plates were incubated for 72 h and culture medium and dressings then removed. Toluidine blue was added to stain the remaining keratinocytes. Following removal of the dye, the plates were photographed under standard conditions and these digital images were analysed using image analysis software. Data was analysed using Student's t-test. In the present study, Silvazine is the most cytotoxic agent. Seventy-two hour exposure to Silvazine in the present study results in almost no keratinocyte survival at all and a highly statistically significant reduction in cell survival relative to control, Acticoat and Flamazine (P<0.001, P<0.01, P<0.01, respectively). Flamazine is associated with a statistically significant reduction in cell numbers relative to control (P<0.05), but is much less cytotoxic than Silvazine (P<0.005). In this in-vitro study comparing Acticoat, Silvazine and Flamazine, Silvazine shows an increased cytotoxic effect, relative to control, Flamazine and Acticoat. An in-vivo study is required to determine whether this effect is carried into the clinical setting.


Acetylcholine receptor. Binding properties and ion permeability response after covalent attachment of the local anaesthetic quinacrine.
Membrane vesicles rich in nicotinic acetylcholine receptor prepared from Torpedo californica electric tissue have been irreversibly modified with quinacrine mustard, an alkylating derivative of the local anaesthetic quinacrine. The reaction blocked the ion channel regulated by the acetylcholine receptor. Acetylcholine still bound to the modified membrane vesicles with KD approx. 10(-8). The number of binding sites was reduced by up to 50%. Stopped-flow experiments showed that in contrast to what had been found with the reversibly binding quinacrine no fluorescence changes caused by energy transfer from the irradiated protein to the fluorescent local anaesthetic occurred after addition of agonist. This indicates that the conformational changes associated with the activation of the ion channel are blocked by the covalent reaction with quinacrine mustard. Analysis of the membrane vesicles by SDS-polyacrylamide gel electrophoresis showed that all polypeptide chains assumed to be part of the receptor complex had reacted with the mustard. Even small components, probably lipids, migrating with the dye front, showed fluorescence.


Oleate ameliorates palmitate-induced reduction of NAMPT activity and NAD levels in primary human hepatocytes and hepatocarcinoma cells.
Nicotinamide phosphoribosyltransferase (NAMPT) and nicotinamide adenine dinucleotide (NAD) levels are crucial for liver function. The saturated fatty acid palmitate and the unsaturated fatty acid oleate are the main free fatty acids in adipose tissue and human diet. We asked how these fatty acids affect cell survival, NAMPT and NAD levels in HepG2 cells and primary human hepatocytes. HepG2 cells were stimulated with palmitate (0.5mM), oleate (1mM) or a combination of both (0.5mM/1mM) as well as nicotinamide mononucleotide (NMN) (0.5 mM) or the specific NAMPT inhibitor FK866 (10nM). Cell survival was measured by WST-1 assay and Annexin V/propidium iodide staining. NAD levels were determined by NAD/NADH Assay or HPLC. Protein and mRNA levels were analysed by Western blot analyses and qPCR, respectively. NAMPT enzyme activity was measured using radiolabelled 14C-nicotinamide. Lipids were stained by Oil red O staining. Palmitate significantly reduced cell survival and induced apoptosis at physiological doses. NAMPT activity and NAD levels significantly declined after 48h of palmitate. In addition, NAMPT mRNA expression was enhanced which was associated with increased NAMPT release into the supernatant, while intracellular NAMPT protein levels remained stable. Oleate alone did not influence cell viability and NAMPT activity but ameliorated the negative impact of palmitate on cell survival, NAMPT activity and NAD levels, as well as the increased NAMPT mRNA expression and secretion. NMN was able to normalize intracellular NAD levels but did not ameliorate cell viability after co-stimulation with palmitate. FK866, a specific NAMPT inhibitor did not influence lipid accumulation after oleate-treatment. Palmitate targets NAMPT activity with a consequent cellular depletion of NAD. Oleate protects from palmitate-induced apoptosis and variation of NAMPT and NAD levels. Palmitate-induced cell stress leads to an increase of NAMPT mRNA and accumulation in the supernatant. However, the proapoptotic action of palmitate seems not to be mediated by decreased NAD levels.


[The MRI of the occult cervical metastasis with malignant tumor of the head and neck].
To detect the occult cervical metastasis with MRI so as to find out its diagnostic accurate rate. 41 lateral necks(negative by clinical palpation) with malignant tumor of head and neck were examined by MRI before operation and correlated with postoperative pathologic finding. With MRI evaluation occult cervical metastasis has higher accurate rate(75%), nine of 12 lateral metastatic positive necks were detected on MRI, other three false negative necks were not found, owing to no change of the shape of the metastatic nodes. At present, MRI is one of the effective methods evaluating occult cervical metastasis with malignant tumor of head and neck, but it has still a certain limitation.


Chemotherapy-induced sclerosing cholangitis.
To review the computed tomography (CT), magnetic resonance imaging (MRI) and cholangiographic findings of chemotherapy-induced sclerosing cholangitis (CISC). Between January 1995 and December 2004, 11 patients in the endoscopic retrograde cholangiography database were identified with CISC. Twelve CT, four MRI, 69 endoscopic and nine antegrade cholangiographic studies in these patients were reviewed. Serial change in appearance and response to endoscopic treatment were recorded. CISC showed segmental irregular biliary dilatation with strictures of proximal extrahepatic bile ducts. The distal 5cm of common bile duct was not affected in any patient. CT and MRI findings included altered vascular perfusion of one or more liver segments, liver metastases or peritoneal carcinomatosis. Biliary strictures needed repeated stenting in 10 patients (mean: every 4.7 months). Cirrhosis (n=1) or confluent fibrosis (n=0) were uncommon findings. CISC shares similar cholangiographic appearances to primary sclerosing cholangitis (PSC). Unlike PSC, biliary disease primarily involved ducts at the hepatic porta rather than intrahepatic ducts. Multiphasic contrast-enhanced CT or MRI may show evidence of perfusion abnormalities, cavitary liver lesions, or metastatic disease.


The logic of a controversy: the case of Agent Orange in Australia.
Since 1979 the Vietnam Veterans' Association of Australia (VVAA) has claimed that exposure to the herbicide Agent Orange in Vietnam has adversely affected the health of Vietnam veterans and their families. A campaign for government recognition of diseases and disabilities caused by herbicide exposure led in 1983 to the appointment of the Evatt Royal Commission which, after a 2-year inquiry, comprehensively rejected the VVAA's claim. The Evatt Commission's findings have not been accepted by the VVAA and the claim continues to be defended, albeit in a highly qualified form. This controversy exemplifies the way in which a claim can attract public support, and persist despite rejections by Committees of Inquiry. An understanding of the reasons for the persistence of controversy requires an understanding of the logic of rejecting causal claims, the psychology of everyday inductive reasoning, and the interaction between politics and science.


Retrograde labeling of migrating spinal motoneurons in bullfrog larvae.
Horseradish peroxidase (HRP) was applied to the ventral root or developing limb bud of bullfrog tadpoles. In animals younger than Stage XII/XIII [9] ventral root application consistently labeled neurons located between the lateral motor column (LMC) and the neuroepithelium. Only occasional motoneurons outside the LMC were labeled in animals older than Stage XIV/XV. Limb bud application of HRP in any-stage tadpole did not label ectopic LMC motoneurons. These results indicate that LMC motoneurons have sent their axons out the ventral root before completion of migration but that these axons may not as yet have reached the limb bud.


Arthroscopy with open surgery for treatment of synovial chondromatosis of the temporomandibular joint.
We report a case of synovial chondromatosis of the temporomandibular joint in which both joint compartments were affected. Because of the important involvement of the medial aspect of the joint, arthrotomy was done with arthroscopic assistance.


Transformation under pressure: Discovery of a novel crystalline form of anthelmintic drug Praziquantel using high-pressure supercritical carbon dioxide.
Supercritical carbon dioxide (CO2) has been used as a processing technique to control polymorphism of pharmaceuticals. However, there are fewer reports of novel polymorphs being discovered by supercritical CO2 processing. As supercritical crystallization methods gain attention for potential in pharmaceutical processing, they may become a critical screening tool for discovery of new polymorphs. In this work, a case study is presented for a novel crystalline form of the anthelmintic drug, Praziquantel, found through supercritical CO2 processing. The novel form of Praziquantel was characterized by chromatography, nuclear magnetic resonance and infrared spectroscopy, X-ray powder diffraction, thermal analysis, and scanning electron microscopy. Furthermore, the novel form exhibited 13-20% improved solubility compared to commercial Form A between pH 1.6 and 7.5 and was physically stable under stressed conditions (40 °C and 75% relative humidity) for 7.5 weeks. Overall, this work showed that supercritical CO2 processing is a valuable tool to screen for novel, and possibly viable polymorphs of pharmaceutical compounds with improved properties.


Oxylipin profile and antioxidant status of potato tubers during extended storage at room temperature.
Potato tubers (cv. Bintje) (Solanum tuberosum L.) were stored under extreme conditions at 20 degrees C for 350 days without sprout inhibitors in order to assess whether aging- and/or senescence-related processes occurred. Under these extreme storage conditions, multiple sprouting followed by the formation of daughter tubers occurs. At the same time, an increase in respiration intensity, as evidenced by cytochrome c oxidase activity (E.C. 22.214.171.124), is observed, leading to a potential increase in reactive oxygen species (ROS) production. As polyunsaturated fatty acids are priority targets of oxidative attacks, the damage to lipids was assessed by oxylipin profiling in both free and esterified forms. Oxylipin profiling showed a predominance of linoleic acid-derived oxylipins and of 9-hydroxy and 9-hydroperoxy fatty acids in both free and esterified forms. No significant accumulation of individual oxylipin was observed 350 days after harvest. To further understand the absence of lipid breakdown products accumulation, the main enzymatic and non-enzymatic antioxidants were assessed. Antioxidant enzyme activities [superoxide dismutase (E.C. 188.8.131.52), catalase (E.C. 18.104.22.168.), ascorbate peroxidase (E.C. 188.8.131.52)] were enhanced during the advanced phase of aging. The main non-enzymatic antioxidant compound, ascorbate, decreased markedly in the early stages of storage, followed by a slower decline. Total radical scavenging activity was also maintained at the end of the storage period. Our results indicate that the enhanced aging process occurring during storage at room temperature does not seem to be associated with the changes classically encountered during leaf senescence or seed aging and that the observed degenerative processes do not surpass the protective potential of the tubers.


The visual system supports online translation invariance for object identification.
The ability to recognize the same image projected to different retinal locations is critical for visual object recognition in natural contexts. According to many theories, the translation invariance for objects extends only to trained retinal locations, so that a familiar object projected to a nontrained location should not be identified. In another approach, invariance is achieved "online," such that learning to identify an object in one location immediately affords generalization to other locations. We trained participants to name novel objects at one retinal location using eyetracking technology and then tested their ability to name the same images presented at novel retinal locations. Across three experiments, we found robust generalization. These findings provide a strong constraint for theories of vision.


Substrate balances across the forearm during the early postoperative period.
The substrate metabolism of skeletal muscle has been studied by the forearm technique in 23 patients after abdominal operation. The data were compared to those of 55 healthy volunteers after an overnight fast. While the uptake of glucose into forearm muscle tissue was increased in relation to its elevated arterial concentration, its rate of oxidation was clearly reduced and the muscle extraction of ketone bodies was increased. Simultaneously, the production of lactate was doubled and the output of glycerol and alanine significantly enhanced. From these data one may conclude that 3 hours after abdominal surgery the energy requirements of skeletal muscle tissue are adequately met by the substrates provided that glucose sparing occurs in association with a rising arterial concentration of KB, and that the release of alanine is accelerated in spite of abundant caloric supply.


Chromium (VI) induced oxidative damage to DNA: increase of urinary 8-hydroxydeoxyguanosine concentrations (8-OHdG) among electroplating workers.
To investigate the concentration of urinary 8-hydroxydeoxyguanosine (8-OHdG) among electroplating workers in Taiwan. Fifty workers were selected from five chromium (Cr) electroplating plants in central Taiwan. The 20 control subjects were office workers with no previous exposure to Cr. Urinary 8-OHdG concentrations were determined using high performance liquid chromatography with electrochemical detection. Urinary 8-OHdG concentrations among Cr workers (1149.5 pmol/kg/day) were higher than those in the control group (730.2 pmol/kg/day). There was a positive correlation between urinary 8-OHdG concentrations and urinary Cr concentration (r = 0.447, p < 0.01), and urinary 8-OHdG correlated positively with airborne Cr concentration (r = 0.285). Using multiple regression analysis, the factors that affected urinary 8-OHdG concentrations were alcohol, the common cold, and high urinary Cr concentration. There was a high correlation of urinary 8-OHdG with both smoking and drinking, but multiple regression analysis showed that smoking was not a significant factor. Age and gender were also non-significant factors. 8-OHdG, which is an indicator of oxidative DNA damage, was a sensitive biomarker for Cr exposure.


Receptors for thymosin alpha 1 on mouse thymocytes.
Thymosin alpha 1 is able to act in vitro to stimulate T-cell precursors and to induce surface markers. The initial mechanism by which alpha 1 activates T cells could be the binding of alpha 1 to cell membranes. Using a specific anti-alpha 1 antibody and an indirect immunofluorescence procedure it was found that thymosin alpha 1 binds to the surface of a large portion of murine lymphocytes. Furthermore, thymocytes have been fractionated into immature and mature subpopulations by using the peanut agglutinin (PNA) technique. It was found that PNA+, immature cells showed specific receptors for alpha 1 on the cell membrane.


Kartogenin Combined Platelet-Rich Plasma (PRP) Promoted Tendon-Bone Healing for Anterior Cruciate Ligament (ACL) Reconstruction by Suppressing Inflammatory Response Via targeting AKT/PI3K/NF-κB.
Anterior cruciate ligament (ACL) rupture is the most common sports injuries and PRP has the potential to be a kartogenin (KGN) carrier to promote collagen fibril organization and cartilage regenerative in the tendon-bone interface. This paper aimed to investigate co-injection of KGN-PRP into the bone tunnels of ACL reconstructions which could enhance tendon-bone healing graft osteointegration effectively. HPLC was used to measured release rate of KGN from KGN-PRP gel. Then, an ACL injury reconstruction model in rabbits was established and the rabbits received saline, PRP, and KGN-PRP injection onto the tendon-bone interface after reconstruction. The tissue was harvested from the tendon-bone interface at 4 weeks and 8 weeks post-surgery, and the sections were stained with Safranin O/fast green to detected tendon-bone healing. Immunochemistry staining was used to analyze VEGF, collagen I, and HIF-1α expression, and ELISA assay was used for detecting IL-6, TNF-α, and COX-2 concentrations. The expression levels of AKT/PI3K/NF-κB-related protein and mRNA were presented by Western blot and qPCR. The release rate of KGN was high within 4 h of KGN-PRP gel and followed by a slow release until 7 days. The Safranin O/fast green staining results indicated that tendon-bone interface in sham and mock group existed gap and tissue disorganization. The KGN + PRP group showed the positive color of the healing interface was more obvious and cartilage tissue began to be generated in large amounts at this interface. The maximum tensile force of KGN-PRP injection tendon-bone healing site was significantly higher than that of PRP group, and KGN-PRP effectively promoted fibro chondrogenesis and tendon-bone healing. The expression of collagen I, VEGF, and HIF-1α in regenerated tissues at the healing interface was significantly increased by KGN-PRP treatment compared with the mock and sham groups. The expressions of IL-6, TNF-α, and COX-2 after KGN-PRP treatment were significantly decreased in tendon-bone interface compared to the mock group. WB and qPCR results showed KGN-PRP treatment effectively inhibits AKT/PI3K/NF-κB activation of inflammatory pathways, thereby reducing the level of inflammation to promote wound healing. PRP is an effective carrier for KGN with the sustained release of KGN. After ACL reconstruction, injection of KGN-PRP gel significantly reduced the inflammatory response and inhibited AKT/PI3K/NF-κB activation in cartilage tissue, which promoted tendon-bone healing.


Safety and tolerability of subcutaneous PTHrP(1-36) in healthy human volunteers: a dose escalation study.
Parathyroid hormone-related protein (PTHrP) is an anabolic skeletal agent in mice, rats and humans. In previous studies, we have demonstrated that PTHrP can be administered to osteoporotic postmenopausal women at a dose of 6.56 microg/kg/day (or approximately 400 microg/day) for 3 months to yield a 4.7% increase in lumbar spine BMD. This regimen was free of hypercalcemia or adverse effects. Moreover, PTHrP appeared to stimulate bone formation selectively, without stimulating bone resorption. This efficacy in the absence of adverse effects, as well as the apparent "pure anabolic" action of PTHrP, prompted us to attempt to define the complete therapeutic window for PTHrP. In this study, we gradually escalated the dose of PTHrP(1-36) from 9 to 28 microg/kg (or approximately 570 microg to 1,946 microg) administered as a single subcutaneous dose to 22 healthy young adult subjects. PTHrP(1-36) was well tolerated even at the highest dose, just under 2.0 mg, some five times higher than we have previously demonstrated to be effective in increasing bone mass, and some 100 times higher than the maximal approved dose of PTH(1-34). Despite the large dose of PTHrP, the highest serum calcium achieved was 10.6 mg/dl, and this was observed in only one subject at the highest dose. The mean serum calcium in subjects receiving the highest dose was 9.6 mg/dl. Only one subject experienced adverse symptoms/signs, and this was at the highest dose. We conclude that subcutaneous PTHrP(1-36) is safe when administered in single doses approaching 2.0 mg. These findings indicate that the therapeutic window for PTHrP(1-36) in humans is wide and permit the design and implementation of longer safety and efficacy trials.


Carbon-carbon bond formation involving reactions of alkynes with group 9 metals (Ir, Rh, Co): preparation of conjugated olefins.
Stable alkynyl complexes of iridium(III) (L(n)Ir-triple bond-R) that are prepared from the reactions of terminal alkynes readily undergo the intramolecular C-C bond-forming reactions between the alkynyl and adjacent hydrocarbyl ligands to yield conjugated olefins. These reactions are initiated by electrophiles (H(+), Me(+)) that attack the beta carbon of the alkynyl ligand to increase the electrophilicity of the alpha carbon of the alkynyl ligand. The C-C bond is then formed between the alpha carbons of the alkynyl and adjacent hydrocarbyl ligands.


Genetic polymorphisms and possible gene-gene interactions in metabolic and DNA repair genes: effects on DNA damage.
We investigated in a central European population, the association between genetic polymorphisms in several genes coding for xenobiotic metabolizing enzymes (CYP1A1, CYP2E1, EPHX1, GSTP1, GSTM1 and GSTT1) and in DNA repair genes (XPD, XPG, XPC and XRCC1) and the levels of single-strand breaks (SSBs) and SSB endonuclease III sensitive sites (endoIII sites) in peripheral blood lymphocytes. No significant differences in the mean levels of SSBs and endoIII sites after stratification for main confounders and occupational exposure were observed in the studied population. Significantly higher levels of SSBs were observed in individuals bearing the wild-type alleles (AA) (0.75+/-0.51SSB/10(9)Da) and heterozygous (AC) genotypes (0.67+/-0.49SSB/10(9)Da) compared to those with homozygous XPD (CC) genotype (0.43+/-0.28SSB/10(9)Da, P=0.033). A moderate increase in the levels of SSBs was also found in individuals with the homozygous XPG exon 15 wild type (GG) and heterozygous (GC) genotypes in comparison to those with the homozygous (CC) genotype (P=0.066) and in individuals with low activity EPHX1 genotype in comparison to those with high activity genotype. Nevertheless, these differences were not statistically significant. No other significant association was found. When gene-gene interactions were evaluated, a combination of EPHX1 activity genotypes with that of either XPD or XPG significantly (P=0.003 and 0.016, respectively) modulated SSB levels resulting in a three-fold difference between the "protective" and the "adverse" genotype-combinations. Almost three-fold differences in SSB levels were found between the "protective" and the "adverse" genotype-combinations of EPHX1 activity genotype and GSTM1 or GSTT1 genotypes, respectively. In conclusion, our results suggest a relation between markers of genotoxicity and polymorphisms in genes coding for xenobiotic metabolizing and DNA repair enzymes as well as a modulating effect of combinations of these polymorphisms.


Seasonal variation of blood pressure in children.
Seasonal blood pressure (BP) variation is mostly found between the summer and winter months. Guidelines for diagnosis and treatment of hypertension in children have not considered this variation until recently. This review aims to present an overview of seasonal BP variation in childhood along with potential underlying pathophysiological mechanisms and long-term implications as well as conclusions for future studies. In pediatric cohorts, seven studies investigated seasonal changes in BP. These changes amount to 3.4-5.9 mmHg (or 0.5-1.5 mmHg per - 1 °C difference in environmental temperature) in systolic BP with a peak in fall or winter. Potential mechanisms and mediators of seasonal BP variation include sympathetic activation of the nervous system with an increase of urinary and plasma norepinephrine levels in the winter season. Additionally, the physical activity among children and adolescents was inversely correlated with BP levels. Temperature sensitivity of BP and pediatric BP levels predict future systolic BP and target-organ damage. Therefore, cardiovascular events may even be long-term complications of seasonal BP variation in pediatric hypertensive patients. Overall, these data strongly suggest an important effect of ambient temperature on BP in children. Additional studies in pediatric cohorts are needed to define how best to incorporate such variation into clinical practice.


Neuropeptide Y receptor-specifically modulates human neutrophil function.
Despite a continuously growing body of evidence highlighting the role of NPY in the immune system, surprisingly little is known about its ability to alter human leukocyte function. We therefore set out to examine NPY receptor expression and functional effects of NPY in freshly isolated human neutrophils. Our results not only demonstrate for the first time the presence of specific NPY receptors on human neutrophils, but also unveil of how these receptors differentially modulate critical functions of neutrophils such as phagocytosis of bacteria as well as the release of reactive oxygen species.