Theoretical prediction of gas-phase infrared spectra of imidazo[1,2-a]pyrazinediones and imidazo[1,2-a]imidazo[1,2-d]pyrazinediones derived from glycine.
Imidazo[1,2-a]pyrazine-3,6-diones and imidazo[1,2-a]imidazo[1,2-d]pyrazine-3,8-diones can be produced by pyrolysis of simple amino acids. While such bicyclic and tricyclic amidines were detected and characterized by IR spectroscopy for some alpha-substituted amino acids, the parent systems composed of glycine fragments are unknown up to now. IR spectra for five amidines derived from glycine were calculated by using different semi-empirical (PM3, AM1, MNDO and MINDO/3), HF, and hybrid DFT (B3LYP, B3P86 and B3PW91) methods in conjunction with 6-31G(d) basis set (for HF and DFT). Vibration frequencies in the experimental IR spectra were predicted based upon the B3LYP data, by correcting the calculated wavenumbers by a scaling factor of 0.959. The behavior of most characteristic bands (nu(CX), nu(NH), etc.) and their shifts with respect to such bands in the spectra of alanine and alpha-aminoisobutyric acid derivatives studied before, are discussed. Performance of the semi-empirical methods was tested, bearing in mind possible future needs for IR spectra predictions for larger molecular systems of similar chemical nature; the use of MINDO/3 and MNDO is recommended. A basis set effect on the B3LYP fundamental vibration frequencies for hexahydroimidazo[1,2-a]pyrazine-3,6-dione was studied by varying Pople basis sets from minimal STO-3G to 6-311++G(d, p). No significant improvements were found beyond the 6-31G(d) basis set, which thus can be recommended to predict IR spectra for the amidines and similar molecules.


Large yellow croaker (Larimichthys crocea) mitofusin 2 inhibits type I IFN responses by degrading MAVS via enhanced K48-linked ubiquitination.
In mammals, mitofusin 2 (MFN2) is involved in mitochondrial fusion, and suppresses the virus-induced RIG-I-like receptor (RLR) signaling pathway. However, little is known about the function of MFN2 in non-mammalian species. In the present study, we cloned an MFN2 ortholog (LcMFN2) in large yellow croaker (Larimichthys crocea). Phylogenetic analysis showed that MFN2 emerged after the divergence of amphioxus and vertebrates. The protein sequences of MFN2 were well conserved from fish to mammals. LcMFN2 was expressed in all the tissues/organs examined at different levels, and its expression was upregulated in response to poly(I:C) stimulation. Overexpression of LcMFN2 inhibited MAVS-induced type I interferon (IFN) promoter activation and antiviral gene expression. In contrast, knockdown of endogenous LcMFN2 enhanced poly(I:C) induced production of type I IFNs. Additionally, LcMFN2 enhanced K48-linked polyubiquitination of MAVS, promoting its degradation. Also, overexpression of LcMFN2 impaired the cellular antiviral response, as evidenced by the increased expression of viral genes and more severe cytopathic effects (CPE) in cells infected with spring viremia of carp virus (SVCV). These results indicated that LcMFN2 inhibited type I IFN response by degrading MAVS, suggesting its negative regulatory role in cellular antiviral response. Therefore, our study sheds a new light on the regulatory mechanisms of the cellular antiviral response in teleosts. The online version contains supplementary material available at 10.1007/s42995-023-00189-8.


Gefitinib resistance in HCC mahlavu cells: upregulation of CD133 expression, activation of IGF-1R signaling pathway, and enhancement of IGF-1R nuclear translocation.
Hepatocellular carcinoma (HCC) is the major form of primary liver cancer which accounts for more than half million deaths annually worldwide. While the incidence of HCC is still on the rise, options of treatment are limited and the overall survival rate is poor. The acquisition of cancer drug resistance remains one of the key hurdles to successful treatment. Clearly, a thorough understanding of the underlying mechanisms is needed for new strategies to design novel treatments and/or to improve the current therapies. In the present study, we examined the expression of cancer stem cell (CSC) marker CD133, the activation of insulin-like growth factor 1 receptor (IGF-1R) signaling, and the nuclear translocation of IGF-1R in HCC Mahlavu cells under the treatment of gefitinib, a cancer drug that inhibits epidermal growth factor receptor (EGFR) pathway. Our results demonstrated that Mahlavu cells exhibited strong gefitinib resistance and the CD133 expression level was dramatically increased (from 3.88% to 32%) after drug treatment. In addition, the gefitinib treated cells displayed increased levels of phosphorylation in IGF-1R and Akt, indicating the intensified activation of this cancer-associated signaling pathway. Moreover, we revealed that IGF-1R underwent nuclear translocation in gefitinib treated cells using confocal microscopy. The IGF-1R nuclear translocation was enhanced under gefitinib treatment and appeared in a dose-dependent manner. Our findings suggest that increased IGF-1R nuclear translocation after gefitinib treatment may contribute to the drug resistance and IGF1-R activation, which might also associate with the upregulation of CD133 expression.


The involvement of post-translational modifications in cardiovascular pathologies: Focus on SUMOylation, neddylation, succinylation, and prenylation.
Cardiovascular disease (CVD) is one of the most threatening diseases to human health and life, and the number of patients is increasing year by year. Thus, it is of great significance to study the pathogenesis, prevention and treatment of CVDs. The occurrence and development of CVDs involve dynamic, complex and delicate intracellular processes and the pathogenesis is not entirely clear. In contrast to genetic mutations, most of the protein post-translational modifications (PTMs) are reversible, and can affect the activity, stability, subcellular localization, protein-protein interaction etc., of the substrate targets, emerging as key mediators of a number of CVD progression. Under pathological conditions, the PTMs undergo aberrant balances which cause changes of the substrate target proteins in expression level, localization and capacity to activate downstream signaling pathways. Therefore, new approaches can be created aiming to correct the abnormal PTM alterations in treating CVDs. This review summarizes some of the more recent advances in PTMs, focusing on SUMOylation, neddylation, succinylation, and prenylation, and the effect of these modifications on cardiovascular function and progression, which may provide potential targets for future therapeutics.


Dose-escalation of oxaliplatin in hemodialysis patient treated with FOLFOX therapy: A case report.
Oxaliplatin is a key part of the standard treatment for colorectal cancer which is formally contraindicated in patients with severe renal dysfunction. Here, we investigated a safe and efficient dosing schedule of oxaliplatin in folinic acid, fluorouracil, and oxaliplatin (FOLFOX) regimen by monitoring total and free platinum concentrations in plasma. A 47-year-old female with chronic hemodialysis was diagnosed with left-sided colon cancer and underwent colectomy. One year later, she was presented with omentum metastasis and needed further treatment. The computed tomography (CT) scanning revealed multiple omental nodules. Positron emission tomography-CT (PET-CT) showed increased uptake of the nodules. The patient was treated with FOLFOX therapy every 3 weeks. The oxaliplatin began with 50 mg/m and gradually increased 85 mg/m as in the standard regimen. A 4-hour dialysis was started 1 hour after the end of oxaliplatin infusion. The free platinum concentration time curve showed a biomodel pattern. The Cmax of the 1st peak we observed in our patients at the standard dose is comparable to patients with normal renal function. This patient was treated with FOLFOX for 12 courses. No apparent adverse effect was observed during the treatment. The FOLFOX can be safely administered in hemodialysis patients on a long-term basis. Dose reduction of oxaliplatin is not necessarily needed if hemodialysis is performed soon after the infusion. Further studies are needed to distinguish between active and inactive oxaliplatin products during the 2nd peak of the free platinum concentration curve in this population.


Quantitation of striatal and extrastriatal D-2 dopamine receptors using PET imaging of [(18)F]fallypride in nonhuman primates.
[(18)F]Fallypride is a highly selective, high-affinity dopamine D-2 receptor ligand. The high affinity, K(D) = 30 pM, makes it a suitable candidate for visualizing both striatal and extrastriatal binding in the brain. In this work, dynamic PET studies of two macaque monkeys were acquired along with arterial plasma samples. Compartmental analysis and Logan plots were used to analyze the striatum, thalamus, frontal, and temporal cortices and to validate a reference region of analysis which yields a distribution volume ratio (DVR). The cerebellum was used as the reference region. The results indicate that all methods of analysis are in close agreement over all the analyzed regions in the brain. The average DVRs for the two monkeys was found to be: caudate = 26, putamen = 29, thalamus = 3.8, frontal ctx = 1.7, and temporal ctx = 1.7 on a high-resolution PET scanner. It was found that a scan time of 2 h is needed to accurately estimate the DVR for all regions of the brain. The striatal regions require the longest to linearize and are the most sensitive to variations in the average tissue-to-plasma efflux constant, k(2). For the extrastriatal regions, the effect of the k(2) term on DVR calculation is negligible. Repeatability measurements for all regions were found to be within 10% using the DVR parameter.


Genetic susceptibility to head and neck squamous cell carcinoma.
Head-and-neck squamous cell carcinoma (HNSCC) is the sixth most common cancer worldwide, and its incidence is growing. Although environmental carcinogens and carcinogenic viruses are the main etiologic factors, genetic predisposition obviously plays a risk-modulating role, given that not all individuals exposed to these carcinogens experience the disease. This review highlights some aspects of genetic susceptibility to HNSCC: among others, genetic polymorphisms in biotransformation enzymes, DNA repair pathway, apoptotic pathway, human papillomavirus-related pathways, mitochondrial polymorphisms, and polymorphism related to the bilirubin-metabolized pathway. Furthermore, epigenetic variations, familial forms of HNSCC, functional assays for HNSCC risk assessment, and the implications and perspectives of research on genetic susceptibility in HNSCC are discussed.


The early phyllosoma stages of spiny lobster Panulirus echinatus Smith, 1869 (Decapoda: Palinuridae) reared in the laboratory.
The early stages of the Panulirus echinatus were hatched and reared in the laboratory. Ovigerous females were captured in their habitat and carefully transported to the laboratory. Larvae were transferred in a recirculation water tank at a density of 10 larvae.L(-1). The larvae were fed on Artemia and gonads of mussel Brachydonts sp. Microalgae Dunaliella viridis was added at a concentration of 150 x 10(4) cell.mL(-1). Larvae and exuviae of each zoeal stage were preserved in an alcohol 70% + glycerin (1:1) solution. The phyllosomas moulted eight times; the intermoulting period of each instar averaged about 7 to 10 days. The main morphological changes of each appendage were described in detail, illustrated and compared with previous reports.


Effects of mirthful laughter on pain tolerance: A randomized controlled investigation.
Chronic pain is a debilitating condition that affects many people. Currently, there is no single treatment known to cure or assure relief from chronic pain. Accordingly, the management of patients' discomfort is an integral part of treating chronic pain. Such treatment, however, is not effective for many patients. We investigated whether mirthful laughter provided by comic relief can influence pain tolerance and muscle soreness in young healthy participants. Forty participants underwent a randomized controlled cross-over designed experiment. Each participant was exposed to a comedy video eliciting mirthful laughter and an uninteresting documentary. Delayed onset muscle soreness was induced in one leg at a time by eccentric exercise. Pain tolerance was tested using blunt force application and assessed subjectively using a visual analog scale. Watching the comedy video elicited a significantly greater irregular breathing pattern compared with watching the documentary video (p < 0.001). After watching the comedy, the participants' positive affect was increased (Δ2 ± 1) while it was largely decreased (Δ-11 ± 2) after watching the documentary video (p < 0.001). Pain tolerance was decreased by 17 ± 5 N after viewing the documentary video (p < 0.001), but did not change significantly after watching the comedy. Thirty minutes of watching a comedy eliciting laughter favorably influenced pain tolerance in healthy humans. CLINICAL TRIAL NO.: #NCT02896075.


Species segregation in one-dimensional granular-system simulations.
We present one-dimensional molecular dynamics simulations of a two-species, initially uniform, freely evolving granular system. Colliding particles swap their relative position with a 50% probability allowing for the initial spatial ordering of the particles to evolve in time and frictional forces to operate. Unlike one-dimensional systems of identical particles, two-species one-dimensional systems of quasi-elastic particles are ergodic and the particles' velocity distributions tend to evolve towards Maxwell-Boltzmann distributions. Under such conditions, standard fluid equations with merely an additional sink term in the energy equation, reflecting the non-elasticity of the interparticle collisions, provide an excellent means to investigate the system's evolution. According to the predictions of fluid theory we find that the clustering instability is dominated by a non-propagating mode at a wavelength of the order 10 pi L/N epsilon , where N is the total number of particles, L the spatial extent of the system and epsilon the inelasticity coefficient. The typical fluid velocities at the time of inelastic collapse are seen to be supersonic, unless N epsilon<or= 10 pi . Species segregation, driven by the frictional force occurs as a result of the strong temperature gradients within clusters which pushes the light particles towards the clusters' edges and the heavy particles towards the center. Segregation within clusters is complete at the time of inelastic collapse.


Schedules of Controlled Substances: Placement of Brivaracetam Into Schedule V. Final rule.
This final rule adopts without change an interim final rule with request for comments published in the Federal Register on May 12, 2016. The Drug Enforcement Administration is placing the substance brivaracetam ((2S)-2-[(4R)-2-oxo-4-propylpyrrolidin-1-yl] butanamide) (also referred to as BRV; UCB-34714; Briviact) (including its salts) into schedule V of the Controlled Substances Act. This scheduling action is pursuant to the Controlled Substances Act, as revised by the Improving Regulatory Transparency for New Medical Therapies Act which was signed into law on November 25, 2015.


Massive hemorrhage during radiofrequency ablation of a pulmonary neoplasm.
Radiofrequency ablation is a new investigational therapy for primary and secondary neoplasms. In this article, the authors describe the anesthetic management of the radiofrequency ablation of a pulmonary neoplasm complicated by massive hemorrhage.


Increased oxidative stress in subjects exposed to carbon disulfide (CS2)--an occupational coronary risk factor.
There is considerable epidemiological evidence that workers exposed to carbon disulfide (CS2) develop premature atherosclerosis leading to increased rates of coronary heart disease (CHD), but mechanisms underlying this association remain obscure. The present study documents that occupational exposure to CS2 modifies the oxidative status of plasma, which is a major determinant of the susceptibility to atherosclerosis. Concentrations of thiobarbituric reactive substances (TBARS), which reflect lipid peroxidation processes in plasma, were determined in 29 men who were exposed to CS2 for more than 20 years, in 24 patients with peripheral atherosclerosis, and in 30 unexposed, healthy control subjects. TBARS concentrations were significantly increased both in CS2-exposed subjects and in patients with peripheral atherosclerosis. Subjects in both groups presented also with decreased levels of plasma alpha-tocopherol, a major plasma antioxidant. In addition, decreased activities of two enzymatic antioxidants, glutathion peroxidase and catalase, were noted both in CS2-exposed subjects and patients with peripheral atherosclerosis. Finally, LDL isolated from both groups showed increased susceptibility to transition metal-induced oxidation in vitro. It is concluded, that occupational exposure to CS2 produces oxidative stress in plasma. This may favor the development of atherosclerosis and increase the incidence of CHD in workers exposed to CS2.


A New Ex Vivo Method for Effective Expansion and Activation of Human Natural Killer Cells for Anti-Tumor Immunotherapy.
Preserving the activities of natural killer (NK) cells in human peripheral blood mononuclear cells (PBMCs) after ex vivo expansion and activation is critical for NK cell-based therapy. Collected from human PBMCs, the NK cells were expanded and activated. The expressions of surface receptors, cytotoxicity against tumor cells, and antibody-dependent cell-mediated cytotoxicity (ADCC) of the NK cells before and after expansion and activation were, respectively, compared. After expansion, the ADCC activity of healthy human NK cells was improved by 32 %, and the cytotoxicity against four types of tumor cells was increased by 19, 29, 26, and 28 %, respectively. The positive expression rates for the activating receptors NKG2D, CD94, NKp46, NKp30, and NKp44 of healthy human NK cells expanded ex vivo were increased by 60, 40, 20, 40, and 63 %, respectively, whereas those for the inhibitory receptors CD158b, NKB1, and NKAT showed no significant changes. The addition of an immunologically active peptide, "TKD," during cell expansion further increased NK cytotoxicity by approximately 10 %. The expanded and activated NK cells from cancer patients achieved average purity which was greater than 90 %, and the cytotoxicity against K562 cells was increased by more than 17 %. Compared with resting NK cells, NK cells both from healthy volunteers and cancer patients expanded and activated ex vivo using our method were significantly more active and demonstrated significantly increased anti-tumor activity. This method could be therefore used as a new and effective approach to meet requirements for anti-tumor immunotherapy.


Complete innervation profile of whole bowel resected at pull-through for Hirschsprung's disease. Unexpected findings.
We used Acetylcholinesterase (AchE) staining and neural cell adhesion molecule (NCAM) immunoreactivity to examine such resected lengths to determine the complete innervation profile of resected bowel in Hirschsprung's disease (HD). Resected specimens of colon obtained at pull-through surgery from 15 patients with HD [short type (S-type: n=5), recto-sigmoid type (RS-type: n=5), long type (L-type: n=5)] were sectioned at 1.5 cm intervals and stained conventionally with AchE histochemistry and NCAM immunohistochemistry. The number of positive nerve fibers (PNFs) in the lamina propria and smooth muscle layers was assessed on a scale of 0 to 3 where 0 meant no PNF and 3 meant many PNFs. The three types of HD had different AchE and NCAM innervation profiles, especially the L-type. There were also different AchE and NCAM innervation patterns seen within the same aganglionic or transitional segments of bowel depending on the site of sampling. The mean proportion of transitional segment in resected specimens from RS-type HD was significantly larger than that of S-type HD (P<0.001) although the proportion of aganglionic segments from S- and RS-type HD were almost the same. Our results suggest that the etiology of L-type HD may be different from the etiology of S- and RS-type HD because of different innervation profiles. Because segments of the excised bowel would appear to have different innervation patterns depending on whether the specimen is sampled proximally or distally, it is important to record the exact site of sampling to allow valid comparisons between types of HD to be made.


Retinal Failure in Diabetes: a Feature of Retinal Sensory Neuropathy.
Physiologic adaptations mediate normal responses to short-term and long-term stresses to ensure organ function. Organ failure results if adaptive responses fail to resolve persistent stresses or maladaptive reactions develop. The retinal neurovascular unit likewise undergoes adaptive responses to diabetes resulting in a retinal sensory neuropathy analogous to other sensory neuropathies. Vision-threatening diabetic retinal neuropathy results from unremitting metabolic and inflammatory stresses, leading to macular edema and proliferative diabetic retinopathy, states of "retinal failure." Current regulatory strategies focus primarily on the retinal failure stages, but new diagnostic modalities and understanding of the pathophysiology of diabetic retinopathy may facilitate earlier treatment to maintain vision in persons with diabetes.


The efficacy and safety of citrate mixture vs sodium bicarbonate on urine alkalization in Chinese primary gout patients with benzbromarone: a prospective, randomized controlled study.
To compare the efficacy and safety of citrate mixture and sodium bicarbonate on urine alkalization in gout patients under benzbromarone treatment. A prospective, randomized, parallel controlled trial was conducted among 200 gout patients in the dedicated gout clinic of the Affiliated Hospital of Qingdao University. The participants were randomly divided into two groups (1:1), sodium bicarbonate group (3 g/day) and citrate mixture group (7 g/day). All patients were prescribed with 25 mg/day benzbromarone at initiation and maintained at a dose of 50 mg/day. Clinical and biochemical data were collected at each follow-up time point (baseline, weeks 2, 4, 8 and 12). A total of 182 patients completed the 12-week urine alkalization study. The urine pH value of both groups increased significantly from the baseline to the final follow-up time point (sodium bicarbonate group, 5.50-6.00, P < 0.05; citrate mixture group, 5.53-5.93, P < 0.05). While the comparisons regarding urine pH between treatment groups showed no significant differences for each time point. The estimated glomerular filtration rate (eGFR) dropped significantly after 12 weeks' trial in the sodium bicarbonate group (P < 0.01), while it was comparable between baseline and the last follow-up (P > 0.05) in the citrate mixture group. Results of urine analysis showed that the incident rate of occult blood in the sodium bicarbonate group was higher than that in the citrate mixture group (38 vs 24%, P < 0.05), accompanied by a similar occurrence of kidney stones. After 12-week follow-up, the frequency of twice gout flare in the citrate mixture group was significantly lower than that in sodium bicarbonate group (4 vs 12%, P = 0.037). No treatment-emergent adverse events occurred. The efficacy of citrate mixture on urine alkalization is comparable to sodium bicarbonate under benzbromarone treatment without significant adverse events. Citrate mixture is superior to sodium bicarbonate in lowering the incidence of urine occult blood and the frequency of gout attacks. Registered with ChiCTR (http://www.chictr.org.cn), No. ChiCTR1800018518.


Unusual findings on cystoscopy in a patient with detrusor overactivity.
Chronic lithium ingestion has been shown to cause polyuria and polydipsia in 20% to 40% of patients, secondary to diabetes insipidus. However, it has not been reported to cause lithium deposition in the bladder. A 77-year-old woman presented to our clinic with complaints of urinary incontinence, urinary urgency, and nocturnal enuresis for the past 3 to 4 years. She denied polydipsia. Her medical history was significant for bipolar disorder for which she had been on lithium therapy for more than 10 years. Metallic deposition was noted in the suburothelium of a urethral biopsy by gross inspection. Suburothelial deposition of metal may act as a bladder irritant and account for this patient's overactive bladder symptoms.


Degradation of myelin proteins by brain endogenous neutral protease.
A proteolytic enzyme isolated from calf brain cytosol, degraded purified myelin basic protein in the presence of Ca2+ at pH 7.0 (Singh, I. and Singh, A.K., Trans. Amer. Soc. Neurochem., 13 (1982) 119). This proteolytic enzyme also degrades basic proteins when incubated with intact myelin in the presence of Ca2+ at a neutral pH. Three hour treatment of purified myelin with this protease resulted in degradation of large basic protein and small basic protein by 73 and 89%, respectively. This proteolytic activity was inhibited by EDTA, leupeptin and low pH (pH 4.0), but was not affected by phenylmethylsulfonylfluoride, p-nitrophenylguanidinobenzoate and pepstatin A. Purified myelin preparations also contain small amounts of Ca2+-activated proteolytic activity. The fact that this is a neutral protease, endogenous to the brain, suggests that it may play a role in the degradation of myelin under physiological and pathological conditions.


Anion-exchange chromatographic behavior of recombinant rat cytochrome b5. Thermodynamic driving forces and temperature dependence of the stoichiometric displacement parameter Z.
The HPLC anion-exchange isocratic retention behavior of the recombinant soluble core of wild type rat cytochrome b5 on Mono Q HR 5/5 was investigated as a function of temperature and sodium chloride concentration at fixed eluent flow-rates. Retention was measured over a range of eluent flow-rates at a specified temperature to determine if true adsorption equilibrium could be approximated by the HPLC method. Apparent Van 't Hoff enthalpies of adsorption obtained from the HPLC retention data were positive, indicating an entropically driven spontaneous adsorption process, and were found to decline with increasing ionic strength. The retention results were interpreted in terms of the stoichiometric displacement model to obtain the apparent number of binding sites in the contact region, Z, as a function of temperature and of protein concentration. Z was found to depend significantly on temperature, even under conditions of nearly complete protein recovery, but did not depend on protein concentration at the low loadings studied.


Methods to detect replication-dependent and replication-independent DNA structure-induced genetic instability.
DNA can adopt a variety of alternative secondary (i.e., non-B DNA) conformations that play important roles in cellular metabolism, including genetic instability, disease etiology and evolution. While we still have much to learn, research in this field has expanded dramatically in the past decade. We have summarized in our previous Methods review (Wang et al., Methods, 2009) some commonly used techniques to determine non-B DNA structural conformations and non-B DNA-induced genetic instability in prokaryotes and eukaryotes. Since that time, we and others have further characterized mechanisms involved in DNA structure-induced mutagenesis and have proposed both replication-dependent and replication-independent models. Thus, in this review, we highlight some current methodologies to identify DNA replication-related and replication-independent mutations occurring at non-B DNA regions to allow for a better understanding of the mechanisms underlying DNA structure-induced genetic instability. We also describe a new web-based search engine to identify potential intramolecular triplex (H-DNA) and left-handed Z-DNA-forming motifs in entire genomes or at selected sequences of interest.


Melanoma arising from the mucosal surfaces of the head and neck.
Oral mucosal malignant melanoma is a rare disease. We reviewed 30 years of data from a tumor registry and identified 65 patients who had head and neck melanomas. Two thirds (43) of the 65 patients were identified as male, with the mean age in the sixth decade. Of the 65 patients, only 6 had melanoma that arose from the oropharyngeal mucosa. Of the lesions involving the oral mucosa, each lesion manifested itself as a mass or was associated with symptoms of discomfort; only one third (2) of the lesions were pigmented. The clinician must carefully examine the head, neck, and oral cavity, and any pigmented lesion that is not recognized as a specific entity, such as amalgam tattoo, should be biopsied. The more common presentation of amelanotic malignant melanoma requires a high index of suspicion for masses identified in the mouth and requires biopsy for definitive diagnosis. The prognosis for oral mucosal malignant melanoma is poor.


Effects of the SNAP-25 Mnll variant on hippocampal functional connectivity in children with attention deficit/hyperactivity disorder.
Attention-deficit/hyperactivity disorder (ADHD) is one of the most widespread and highly heritable neurodevelopmental disorders affecting children worldwide. Although synaptosomal-associated protein 25 (SNAP-25) is a possible gene hypothesized to be associated with working memory deficits in ADHD, little is known about its specific impact on the hippocampus. The goal of the current study was to determine how variations in ADHD's SNAP-25 Mnll polymorphism (rs3746544) affect hippocampal functional connectivity (FC). A total of 88 boys between the ages of 7 and 10 years were recruited for the study, including 60 patients with ADHD and 28 healthy controls (HCs). Data from resting-state functional magnetic resonance imaging (rs-fMRI) and clinical information were acquired and assessed. Two single nucleotide polymorphisms (SNP) in the SNAP-25 gene were genotyped, according to which the study's findings separated ADHD patients into two groups: TT homozygotes (TT = 35) and G-allele carriers (TG = 25). Based on the rs-fMRI data, the FC of the right hippocampus and left frontal gyrus was evaluated using group-based comparisons. The corresponding sensitivities and specificities were assessed. Following comparisons between the patient groups, different hippocampal FCs were identified. When compared to TT patients, children with TG had a lower FC between the right precuneus and the right hippocampus, and a higher FC between the right hippocampus and the left middle frontal gyrus. The fundamental neurological pathways connecting the SNAP-25 Mnll polymorphism with ADHD via the FC of the hippocampus were newly revealed in this study. As a result, the hippocampal FC may further serve as an imaging biomarker for ADHD.


Frequency-dependent capacitance of the apical membrane of frog skin: dielectric relaxation processes.
Impedance analysis of the isolated epithelium of frog skin (northern Rana pipiens) was carried out in the frequency range between 0.1 Hz and 5.5 kHz while Na+ transport was abolished. Under these conditions, the impedance is determined almost completely by the dielectric properties of the apical membranes of the cells and the parallel shunt resistance. The modeling of the apical membrane impedance function required the inclusion of dielectric relaxation processes as originally described by. J. Chem. Phys. 9:341-351), where each process is characterized by a dielectric increment, relaxation frequency, and power law dependence. We found that the apical plasma membrane exhibited several populations of audio frequency dielectric relaxation processes centered at 30, 103, 2364, and 6604 Hz, with mean capacitive increments of 0.72, 1.00, 0.88, and 0.29 microF/cm2, respectively, that gave rise to dc capacitances of 1.95 +/- 0.06 microF/cm2 in 49 tissues. Capacitance was uncorrelated with large ranges of parallel shunt resistance and was not changed appreciably within minutes by K+ depolarization and hence a decrease in basolateral membrane resistance. A significant linear correlation existed between the dc capacitance and Na+ transport rates measured as short-circuit currents (Cadc = 0.028 Isc + 1.48; Isc between 4 and 35 microA/cm2) before inhibition of transport by amiloride and substitution of all Na+ with NMDG (N-methyl-D-glucamine) in the apical solution. The existence of dominant audio frequency capacitive relaxation processes complicates and precludes unequivocal interpretation of changes of capacitance in terms of membrane area alone when capacitance is measured at audio frequencies.


In situ population dynamics of bacterial viruses in a terrestrial environment
Predation by bacteriophages is thought to control bacterial numbers and facilitate gene transfer among bacteria in the biosphere. A thorough understanding of phage population dynamics is therefore necessary if their significance in natural environments is to be fully appreciated. Here we describe the in situ population dynamics of three separate phage populations predating on separate bacterial species, living on the surface of field-grown sugar beet (Beta vulgaris var. Amethyst), as recorded over a 9-month period. The distributions of the three phage populations were different and fluctuated temporally in 1996 (peak density, approximately 10(3) PFU g-1). One of these populations, predating on the indigenous phytosphere bacterium Serratia liquefaciens CP6, consisted of six genetically distinct DNA phages that varied in relative abundance to the extent that an apparent temporal succession was observed between the two most abundant phages, PhiCP6-1 and PhiCP6-4.


Inhibition of adipogenesis by Wnt signaling.
Wnts are secreted signaling proteins that regulate developmental processes. Here we show that Wnt signaling, likely mediated by Wnt-10b, is a molecular switch that governs adipogenesis. Wnt signaling maintains preadipocytes in an undifferentiated state through inhibition of the adipogenic transcription factors CCAAT/enhancer binding protein alpha (C/EBPalpha) and peroxisome proliferator- activated receptor gamma (PPARgamma). When Wnt signaling in preadipocytes is prevented by overexpression of Axin or dominant-negative TCF4, these cells differentiate into adipocytes. Disruption of Wnt signaling also causes transdifferentiation of myoblasts into adipocytes in vitro, highlighting the importance of this pathway not only in adipocyte differentiation but also in mesodermal cell fate determination.


An outbreak of salmonellosis amongst holidaymakers in Madeira, July 1988.
In July 1988, 20 of 49 tourists from Northern Ireland became ill with gastrointestinal symptoms while on holiday in Madeira and four were admitted to hospital. Salmonella enteritidis was subsequently isolated from eight members of the party. Epidemiological investigations implicated fried and/or scrambled eggs as the vehicle of infection.


The gamma loop, a model of glomerular hemodynamics.
A model of rubber tubing which imitates the vascular system of the glomerulus shows two flow anomalies: Partially closing down a clamp at the outlet increases perfusion. An increase in input pressure reduces perfusion. These anomalies are used to explain regulation of renal perfusion and autoregulation.


[Effects of therapy, serology, stage and activity of rheumatoid arthritis on lymphocyte stimulation in whole blood].
We utilized a new, whole blood method to study the stimulation of lymphocytes by phytohemagglutinin in patients with rheumatoid arthritis and healthy volunteers. We observed that lymphocytes of patients with rheumatoid arthritis were significantly less stimulated by phytohemagglutinin than lymphocytes of controls (p less than 0.05). The underlying T-cell defect appeared to be present at all stages and activity levels of disease, and in both seropositive and seronegative patients.


Effect of streptolysin S on liposomes. Influence of membrane lipid composition on toxin action.
The effect of the bacterial cytolytic toxin, streptolysin S, on liposomes composed of various phospholipids was investigated. Large unilamellar vesicles containing [14C]sucrose were prepared by reverse-phase evaporation, and membrane damage produced by the toxin was measured by following the release of labeled marker. The net charge of the liposomes had little or no effect on their susceptibility to steptolysin S and the toxin was about equally effective on liposomes composed of phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine and phosphatidylglycerol. Experiments with liposomes composed of synthetic phospholipids showed that the ability of the toxin to produce membrane damage depended on the degree of unsaturation of the fatty acyl chains. The order of sensitivity was C18 : 2 phosphatidylcholine greater than C18: I phosphatidylcholine greater than C18 : 0 phosphatidylcholine = C16 : 0 phosphatidylcholine. Liposomes containing the latter two phospholipids were virtually unaffected by streptolysin S, and experiments with C18 : 0 phosphatidylcholine suggested that toxin activity does not bind to liposomes composed of phospholipids with saturated fatty acyl chains. The inclusion of 40 mol% cholesterol in C16 : 0 phosphatidylcholine and C18 : 0 phosphatidylcholine liposomes made these vesicles sensitive to streptolysin S. Egg phosphatidylcholine liposomes, which were unaffected at 0 degrees C and 4 degrees C became susceptible to the toxin at these temperatures when cholesterol was included. Liposomes composed of C14 : 0 phosphatidylcholine were unaffected by streptolysin S at temperatures below the chain-melting transition temperature (23 degrees C) of this phospholipid, but became increasingly susceptible above this temperature. The results suggest that the fluidity of the phospholipid hydrocarbon chains in the membrane is important in streptolysin S action.


The Sign Effect in Past and Future Discounting.
We compared the extent to which people discounted positive and negative events in the future and in the past. We found that the tendency to discount gains more than losses (i.e., the sign effect) emerged more strongly for future than for past outcomes. We present evidence from six studies (total N = 1,077) that the effect of tense on discounting is tied to differences in the contemplation emotion of these events, which we assessed by measuring participants' emotions while they either anticipated or remembered the event. We ruled out loss aversion, uncertainty, utility curvature, thought frequency, and connection to the future and past self as explanations for this phenomenon, and we discuss why people experience a distinct mixture of emotions when contemplating upcoming events.


Constantimarinum furrinae gen. nov., sp. nov., a marine bacterium isolated from saline volcanic rock aquifer (lava seawater) at Jeju Island, Republic of Korea.
A Gram-stain-negative, aerobic, rod-shaped (0.3-0.5 × 1.0-1.9 µm), non-motile marine bacterium designated as ALE3EIT was isolated from a saline volcanic rock aquifer (lava sea-water) on Jeju Island, Republic of Korea. The 16S rRNA gene sequence analysis revealed that strain ALE3EIT showed high similarity to 'Altibacter lentus' JLT2010T (97.2%), followed by Marixanthomonas ophiurae KMM 3046T (94.5%). Growth was observed at 10-41°C (optimum, 30°C), at pH 6.0-8.5 (optimum, pH 7.5) and at 0.5-8% (optimum, 4.0%) NaCl. The predominant cellular fatty acids were iso-C15:0 (23.5%), iso-C16:0 (10.2%), iso-C16:0 3OH (10.5%), and iso-C17:0 3OH (16.8%). The DNA G + C contents was 40.4 mol%. The major respiratory quinone was MK-6. The major polar lipids were determined to be phosphatidylethanolamine, two unidentified glycolipids, and two unidentified aminolipids. Several phenotypic characteristics such as production of acetoin, activities of arginine dihydrolase and acid phosphatase, and utilization pattern of carbon sources differentiate strain ALE3EIT from 'A. lentus' JLT2010T. Activities of the lipase, trypsin, α-chymotrypsin and gelatinase and utilization pattern of carbon sources differentiate strain ALE3EIT from M. ophiurae KMM 3046T. The genome of strain ALE3EIT is 3.0 Mbp long and its ANI and AAI values against 'A. lentus' JLT2010T were 76.58 and 72.76, respectively, however, AAI values against members in other genera were lower than 72%. The phylogenomic tree inferred by PhyloPhlAn clearly differentiated the strain ALE3EIT together with strain JLT2010T from other genera in the Falvobacteriaceae. This polyphasic taxonomic data indicates that strain ALE3EIT should be identified as a novel species in the genus 'Altibacter', however, the name has not been validated. Therefore, the strain is classified as a novel genus and is proposed as Constantimarinum furrinae gen. nov., sp. nov. The type strain is ALE3EIT (= KCCM 43303T = JCM 33022T).


An effervescence-assisted switchable deep eutectic solvent based liquid-phase microextraction of triazole fungicides in drinking water and beverage.
A new effervescence-assisted switchable deep eutectic solvent-based liquid phase microextraction (EA-SDES-LPME) combined with HPLC-UV was developed for determination of common triazole fungicides in drinking water and beverages, including myclobutanil, flusilazole, hexaconazole and bitertanol. The alternative extraction solvent was prepared with hexafluoroisopropanol and dipropylamine with the merits of time-saving, easy to collect and cost-effectiveness. The SDES can be reversibly switched between hydrophilic and hydrophobic states by pH adjustment. The homogeneous extraction was achieved under the hydrophilic form of SDES, and the bi-phase separation was obtained easily by adjusting pH value to restore the original hydrophobicity. Moreover, the characterization of SDES was investigated by FTIR and 1H NMR. The main variables affecting extraction efficiency were optimized in detail. Under the optimal conditions, the proposed method shows desirable precision (RSDs less than 18.5%) and acceptable recovery (72.6-95.4%). The lower limits of detection and limits of quantitation were found to be in the range of 1-2 μg L-1 and 5-10 μg L-1, respectively. The formation mechanism of SDES and the extraction mechanism for target analytes were investigated by density functional theory. The proposed methodology was simplicity, sensitive, time-saving and successfully applied to determine triazole fungicides in drinking water and beverages, making it an alternative technique for the analysis of trace analytes with satisfactory sensitivity.


A Practical Guide to Genetic Testing in Inherited Heart Disease.
Genetic testing has increasingly been shown to provide critical information regarding the treatment and management of patients with hereditary cardiomyopathies and arrhythmias and is available for a wide variety of conditions. It can provide information regarding arrhythmia risk, lifestyle recommendations, such as exercise avoidance, pharmaceutical therapies, and prognosis. Beyond the proband, genetic testing can be a valuable tool for cascade screening in the family. Genetic testing should be accompanied with genetic counseling, as genetic tests should be accompanied by expert interpretation, support in cascade family evaluation, and psychosocial considerations. Overall, it should be routinely implemented in arrhythmia and cardiomyopathy clinics.


Smad signaling pathways regulate pancreatic endocrine development.
Expansion of the pancreatic endocrine cell population occurs during both embryonic development and during post-natal pancreatic growth and regeneration. Mechanisms of the expansion of endocrine cells during embryonic development are not completely understood, and no clear mechanistic link has been established between growth of the embryonic endocrine pancreas and the islet cell replication that occurs in an adult animal. We found that transforming growth factor-beta (TGF-β) superfamily signaling, which has been implicated in many developmental processes, plays a key role in regulating pancreatic endocrine maturation and development. Specifically, the intracellular mediators of TGF-β signaling, smad2 and smad3, along with their inhibitor smad7, appear to mediate this process. Smad2, smad3 and smad7 were all broadly expressed throughout the early embryonic pancreatic epithelium. However, during later stages of development, smad2 and smad3 became strongly localized to the nuclei of the endocrine positive cells, whereas the inhibitory smad7 became absent in the endocrine component. Genetic inactivation of smad2 and smad3 led to a significant expansion of the embryonic endocrine compartment, whereas genetic inactivation of smad7 led to a significant decrease in the endocrine compartment. In vitro antisense studies further corroborated these results and supported the possibility that interplay between the inhibitory smad7 and the intracellular mediators smad2/3 is a control point for pancreatic endocrine development. These results should provide a better understanding of the key control mechanisms for β-cell development.


[Intestinal microbiota and ageing: A new intervention route?].
Intestinal microbiota (IM) has continued to be the subject in all types of studies and publications. More is known on its different components and functions, as well as the changes that take place in IM through the life cycle, and the role of the factors involved in these changes. The aim of this review is to update the relationship between IM and aging. The presentation in 4 sections: (i)main factors of the human ageing process, underlining those related with gut changes; (ii)conceptual meaning of words like microbiota and other related terms; (iii)to comment on the most current findings as regards the changes in IM that occur in the ageing process, whether arising from the physiology or from disease situations, or other factors (environment, diet, drugs, etc.), and the health-consequences of these changes, and (iv)possibilities of different active positive interventions, with emphasis on diet measures.


Low angle X-ray diffraction studies on stained rat tail tendons.
Low angle X-ray diffraction patterns of rat tail tendons with heavy metal stains added were examined to help clarify the effects of fixation and staining on collagen fibrils. Fixing and staining of rat tail tendon fibers gives an X-ray pattern with an intensified 3.8 nm row and the preservation of most equatorial features found in the native pattern. The presence of the native pattern features suggests the value of fixation in preserving native structure before staining. Staining of rat tail tendon fibers without prior fixation led to the disappearance of the native equatorial features and the appearance of a new broad row line corresponding to a spacing of around 10.0--17.5 nm. This observation suggests that some alteration has taken place in the native structure and may be related to electron microscopic observations of units of 10.0--20.0 nm in collagen fibrils under some disruptive or developmental conditions.


Formation of distinct signalling complexes involving phosphatidylinositol 3-kinase activity with stimulation of epidermal growth factor or insulin-like growth factor-I in human skin fibroblasts.
We recently described a better correlation of DNA synthesis with phosphatidylinositol (PI) 3-kinase than with mitogen-activated protein (MAP) kinase stimulated by insulin-like growth factor (IGF)-1 or epidermal growth factor (EGF) in human skin fibroblasts (Takahashi et al., 1997, Endocrinology 138:741-750). IGF-I-induced PI 3-kinase activation is generally mediated via insulin receptor substrate (IRS)-1, but EGF-induced PI 3-kinase activation is mediated by various signalling molecules such as ErbB3 and c-Cbl in different cells. We therefore investigated the mechanism regulating PI 3-kinase in human skin fibroblasts by comparing complexes involving PI 3-kinase when stimulated by IGF-I or EGF and found that p115 and p105, which were tyrosine-phosphorylated by EGF stimulation and associated with SHP-2, were also associated with the p85 subunit of PI 3-kinase by EGF. Anti-SHP-2 and anti-p85 subunits of PI 3-kinase antibodies did not coprecipitate tyrosine-phosphorylated EGF receptor or ErbB3; in addition, p115 and p105 appeared to be distinct from tyrosine-phosphorylated c-Cbl. Thus, tyrosine-phosphorylated p115 and p105 may provide a novel platform recruiting p85, which may simultaneously bind to SHP-2. In contrast, tyrosine phosphorylation of p115 or p 105 was undetectable by immunoblot with IGF-I stimulation, and PI 3-kinase activity was mediated via IRS-1 phosphorylated with IGF-I stimulation, little of which was associated with SHP-2. Thus, EGF and IGF-I cause formation of a distinct signalling complex which associates with p85 subunit of PI 3-kinase.


Focal cortical dysplasia on magnetic resonance imaging: a case report.
A case report of an 11 years old boy with new onset of a seizure disorder is presented. A computed tomography scan demonstrated a noncalcified, nonenhancing focal region of abnormal cortex. A magnetic resonance imaging scan delineated both an isointense area of abnormally thickened gyri and linear areas of abnormal high signal intensity in the subjacent white matter. A review of the radiologic and pathologic literature suggests that this lesion represents the entity focal cortical dysplasia as described by Taylor, et al. This abnormality is part of a spectrum of disorders including hamartomas (of tuberous sclerosis), focal cortical dysplasia and heterotopias.


Variations in the distribution, frequency, and phenotype of Langerhans cells during the evolution of malignant melanoma of the skin.
We examined the frequency, distribution, and immunophenotype (S-100 protein, CD1) of epidermal Langerhans cells (LC) in the epidermis overlying primary melanoma, and dermal dendritic cells (DC) in the dermis deep to melanoma, and in adjacent normal skin. There is a substantial reduction in S-100+ LC and a lesser decline in CD1+ LC in the epidermis over melanoma. There is a simultaneous increase in the frequency of cells expressing these phenotypes in the dermis deep to tumor. Double-staining studies in progress show coexpression of S-100 and CD1 in most of these cells. The depletion of S-100+/CD1+ DC from the peritumoral epidermis is maximum in deeper (Clark level III-V) and thicker (> 0.76 mm) tumors. The reduction in total DC in peritumoral epidermis is, however, proportionally less than the reduction in S-100+/CD1+ DC owing to an increase in cells that are S-100-/CD1+. The proportion of S-100+/CD1+ dermal DC deep to tumor is similar to that in normal epidermis and dermis, suggesting that there is no increased migration of S-100-/CD1+ DC from tumor-associated epidermis to subjacent dermis. Non-dendritic leukocytes in the dermis deep to tumor were increased in frequency, maximally in the dermis deep to thinner and more superficial melanomas. Most such cells were T lymphocytes with helper/inducer cells predominating. The data presented show substantial alterations in the frequency, distribution, and phenotype of LC/DC as melanoma evolves, alterations that may be critical for development of effective tumor-directed immunity.


Changes in polyphosphoinositides and phosphatidic acid of erythrocyte membranes in diabetes.
We studied metabolic pool size of polyphosphoinositides and phosphatidate of erythrocyte membranes from normal and diabetic subjects using 32P for 20-h incubation, a sufficiently long period to reach isotopic equilibrium between monoesterphosphate bond and gamma-phosphate of ATP. Phosphatidylinositol 4-monophosphate (PtdIns4P), phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) and phosphatidate were the phospholipids labelled. Metabolic pools of individual phospholipids were estimated, based on their proportionate and absolute radioactivity. A significant decline in radioactivity of phosphatidate and PtdIns(4,5)P2 was seen in erythrocytes from the diabetic subjects, indicating suppression of the metabolically labile pool of these two phospholipids. There was no significant change in PtdIns4P radioactivity between the groups. The direct effect of insulin on phosphorylation of polyphosphoinositides and phosphatidate was also evaluated by a short incubation period of erythrocyte membranes with [gamma-32P]-ATP. Added insulin increased the incorporation of 32P into phosphatidate in a dose-dependent manner that reached a steady state at 2 nM. We conclude that the metabolically labile pool size of phosphatidate is decreased and that of polyphosphoinositides is altered in erythrocyte membranes from diabetic patients.


In vivo evaluation of some biophysical parameters of the facial skin of Indian women. Part I: variability with age and geographical locations.
India is a large country (a subcontinent) of about 3.3 million km2 that covers large ranges in latitude and longitude. The last Indian census counted about 1.21 billion of inhabitants of many origins, creating a vast human diversity and skin types, the variability of which having been previously established. The present study aimed at deepening this knowledge through a set of biophysical measurements to describe, along the skin ageing process, the specificities of various Indian subjects living in different geographical locations. A total of 1204 women, aged 18-84 years, of all socio-economic status, were recruited in four Indian cities (Mumbai, Kolkata, Chennai and Delhi). Measurements of face skin colour properties, elastic properties, sebum production, skin pores and microrelief roughness were performed. With regard skin colour, this study indicates, with age, a darkening of very low amplitude that leads to an increased skin colour heterogeneity. In all subjects, at all ages, the ocular region (dark circles) presents a much darker pigmentation than the cheeks, creating a contrast that appears constant at all ages. In addition to an increased skin colour heterogeneity, a progressive alteration of the skin surface relief, increased sizes of skin pores, a loss of skin elasticity and a drop in sebum production, post-menopause, are observed. This study confirms, in Indian women, some skin ageing measurements found on women from other ethnic groups (i.e. sebum, firmness, wrinkles and pores size) and also identifies some Indian specificities: a high and constant contrast between the ocular region and the cheek colour, associated to a very slow darkening effect along the lifespan.


Social participation as a mediator of the relationships of socioeconomic factors and longevity after traumatic spinal cord injury.
Cohort study. Previous research has indicated that socioeconomic factors affect longevity after traumatic spinal cord injury (SCI). Our purpose was to evaluate whether social participation mediates the relationship between socioeconomic factors and survival status after SCI. Medical university in the southeastern United States. Participants (N = 1540) met the following inclusion criteria: traumatic SCI of at least 1-year duration, minimum of 18 years of age, and having residual impairment from SCI. The main outcome measures were a) survival status as of December 31, 2019, identified by the National Death Index (NDI) search, b) socioeconomic status (SES), measured by education, employment status, and family income, and c) participation, measured by marital/relationship status, hours out of bed per day, days leaving home per week, and nights away home during the past year. Thirty nine percent of participants (n = 602) were decreased by the end of 2019. Socioeconomic factors were associated with longevity controlling for demographic, injury characteristics, and health status. However, the association of SES with longevity was mediated by three social participation mobility indicators (hours out of bed, days out of house, and nights away from home), such that SES was no longer significantly related to longevity after inclusion of the participation variables. Although socioeconomic factors are related to longevity, their relationship appears to be mediated by social participation mobility indicators. Intervention studies are needed to address the modifiable factors that may promote longevity, including promoting an active lifestyle.


Insights into the genetic organization of the Bacillus mycoides cryptic plasmids pDx14.2 and pSin9.7 deduced from their complete nucleotide sequence.
Bacillus mycoides, a member of the Bacillus cereus group of bacteria, can be easily distinguished from close species because of colony shape, made by filaments of cells, resembling fungal hyphae, curving clock- or counterclockwise depending on the strain. Two plasmids, one from a strain curving to the right (pDx14.2), the other from a strain curving to the left (pSin9.7), were sequenced and analyzed for gene content and replication mode. Rolling-circle replication modules and mobilization proteins were found, very similar to those of other plasmids of the B. cereus group bacilli, mostly Bacillus thuringiensis living in the same ecosystem, suggesting active plasmid exchange in nature.


The polymorphic human glutathione transferase T1-1, the most efficient glutathione transferase in the denitrosation and inactivation of the anticancer drug 1,3-bis(2-chloroethyl)-1-nitrosourea.
A member of the Theta class of human glutathione transferases (GST T1-1) was found to display the greatest catalytic activity towards the cytostatic drug 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) of the GSTs studied. In this investigation (the most extensive to date), enzymes from four classes of the soluble human GSTs were heterologously expressed, purified, and kinetically characterized. From the 12 enzymes examined, only GST M2-2, GST M3-3 and GST T1-1 had significant activities with BCNU. This establishes that the activity is not a characteristic of a particular class of GSTs. Although GST M3-3 was previously reported to have the greatest activity with BCNU, the current investigation demonstrates that GST M2-2 is equally active and that GST T1-1 has an approximately 20-fold higher specific activity than either of the Mu class enzymes. A more rigorous kinetic analysis of GST T1-1 gave the following parameters with BCNU: a k(cat) of 0.035 +/-0.003s(-1) and a K(M) of 1.0 +/- 0.1mM. The finding that GST T1-1 has the highest activity towards BCNU is significant since GST T1-1 is expressed in the brain, a common target for BCNU treatment. Furthermore, the existence of a GST T1-1 null allele in up to 60% in some populations, may influence both the sensitivity of tumors to chemotherapy and the severity of adverse side-effects in patients treated with this agent.


Effect of lead, zinc, mercury, and copper with and without estrogen on serum vitellogenin level in Magur fish (Clarias batrachus L.).
Injection(s) of lead, zinc, and mercuric acetate decreased the serum vitellogenin content in Magur fish, while cupric acetate failed to cause any change in the vitellogenin level. Estrogen injections on 7th, 8th and 9th d increased the serum vitellogenin level in normal and copper salt treated fish, but were totally ineffective in altering the reduced vitellogenin content in lead, zinc, and mercury salts treated fish. Vitellogenin level almost restored to normal level at 6 week in lead, zinc, and mercury treated fish, and estrogen injections on 37th, 38th, and 39th d enhanced the serum vitellogenin content in all groups.


Comparison of Fasciola hepatica genotypes in relation to their ability to establish patent infections in the final host.
Fasciola hepatica is a common and economically important parasite of sheep and cattle. Although its marked genetic heterogeneity is well recognised, an association between haplotypes and specific phenotypic traits has yet to be identified. Using experimental infections in cattle this study investigated whether a fragment of mitochondrial DNA (coding for cytochrome c oxidase subunit III, transfer RNA histidine and cytochrome b) and 3 nuclear microsatellite loci (Fh15, Fh23 and Fh25) could be used as markers for the parasite's ability to complete its tissue migration and establish in the liver of the final host. While we did not detect any shift in the frequency of the various genotypes in the population of metacercariae used for the infection on the one hand and the flukes collected from the liver on the other, there was an indication that parasites with heterozygous microsatellite alleles may have a selective advantage over homozygote parasites during their migration in the final host.


Long-term benefits of heart rate variability biofeedback training in older adults with different levels of social interaction: a pilot study.
To test whether heart rate variability (HRV) biofeedback training benefits older adults with different social interaction levels. 32 older adults (16 were institutionalized and 16 were not). Both groups received 14 sessions, 15 min, 3 times a week, with half of the individuals receiving HRV biofeedback training and the other half receiving control training. The following parameters were assessed immediately before and after training, and 4.5 weeks after the last session (follow-up period): aerobic conditioning, anthropometric data, emotional scores, and HRV components. Before the training, the institutionalized individuals had higher scores of loneliness (p < 0.01) and depression (p < 0.0001) and lower social touches (p < 0.0001), body mass (p = 0.04), and body fat percentage (p = 0.002) than the non-institutionalized individuals. HRV biofeedback improved symptoms of depression in both groups. HRV improved only in the non-institutionalized group, and loneliness only in the institutionalized group. Lastly, all changes persisted after the follow-up period. HRV biofeedback training was effective in improving symptoms of depression in older adults. Improvement of HRV and loneliness was dependent on the level of social interaction.


Changes in the transverse dimensions by vertical ramus osteotomy after mandibular prognathism correction.
A harmonious face displays not only a good lateral profile but also a pleasant frontal appearance. The purpose of the current study was to evaluate the changes in the transverse dimensions by vertical ramus osteotomy (VRO) in the treatment of mandibular prognathism correction. Twenty patients who underwent mandibular prognathism correction by VRO were included. Lateral and frontal cephalometric radiographs were obtained at the following stages: preoperative (T1), immediately after the surgery (T2), and completion of orthodontic treatment (T3). Three linear measurements (menton, intercondylion, and intergonial distances) and the ramus angle were compared from T1 to T3. A final mean menton setback of 12.2 mm and upward movement of 0.3 mm were noted. The intercondylion and intergonial distances significantly increased by 5.1 and 7.1 mm, respectively. Without significant difference, ramus angles decreased 1.7 degrees in the right side and 0.1 degrees in the left side. The surgical correction of mandibular prognathism using VRO led to an increase in the transverse dimensions.


Ethyl glucuronide and ethyl sulfate in the zebrafish after ethanol exposure.
Ethanol exposure during pregnancy is an important problem and is the cause of fetal alcohol syndrome (FAS) and fetal alcohol spectrum disorder (FASD). The etiology of FAS and FASD can be elucidated using animal models. Recently, a novel model, the zebrafish (Danio rerio), has garnered the interest of researchers. This study confirmed the negative influence of ethyl alcohol (0.5%, 1.5%, and 2.5% v/v) on the development of zebrafish embryos. The observed malformations included pericardial and yolk sac edema, increased body curvature, tail edema, and a decreased embryo hatching rate. The differences in body length, body width, and heart rate were statistically significant. Due to the similarities in the quantity and function of ethanol biotransformation enzymes between zebrafish and mammals, this study investigated the nonoxidative metabolites of ethanol-ethyl glucuronide (EtG) and ethyl sulfate (EtS)-in zebrafish following ethanol exposure. This research confirmed that EtG and EtS concentrations can be measured in zebrafish embryos, and the levels of these metabolites appear to be associated with the ethyl alcohol concentration in the medium.


Discrepancy between GLUT4 translocation and glucose uptake after ischemia.
Low-flow ischemia results in glucose transporter translocation and in increased glucose uptake. After total ischemia in rat heart, we found no increase in glucose uptake. Here we test the hypothesis that total ischemia is associated with decreased activation of GLUT4 despite translocation. Isolated working hearts (n=70, Sprague-Dawley rats) were perfused for 70 min at physiological workload with Krebs-Henseleit buffer containing [2-3H]glucose (5 mmol/l, 0.05 microCi/ml) with either oleate (0.4 mmol/l, 1%BSA) or pyruvate (5 mmol/l, 1%BSA). After 20 min, hearts were subjected to 15 min of total ischemia followed by 35 min of reperfusion. We measured glucose uptake and intracellular free glucose (IFG) using [2-3H]glucose and [14C]sucrose, and determined the distribution of GLUT4 by colocalization immunofluorescence with Na-K ATP-ase. Cardiac power was 10.1 +/- 0.90 mW before ischemia and did not differ between groups. Recovery was the same in both groups (55.7 +/- 24.8%). Glucose uptake did not differ between groups before ischemia, and did not increase during reperfusion. Despite evidence of GLUT4 translocation after reperfusion in both groups, IFG did not increase compared with before ischemia. We conclude that there is a discrepancy between glucose transporter availability and glucose uptake after ischemia, which may be due to inhibition of GLUT4 in the plasma membrane.


A peptide core motif for binding to heterotrimeric G protein alpha subunits.
Recently, in vitro selection using mRNA display was used to identify a novel peptide sequence that binds with high affinity to Galpha(i1). The peptide was minimized to a 9-residue sequence (R6A-1) that retains high affinity and specificity for the GDP-bound state of Galpha(i1) and acts as a guanine nucleotide dissociation inhibitor (GDI). Here we demonstrate that the R6A-1 peptide interacts with Galpha subunits representing all four G protein classes, acting as a core motif for Galpha interaction. This contrasts with the consensus G protein regulatory(GPR) sequence, a 28-mer peptide GDI derived from the GoLoco (Galpha(i/0)-Loco interaction)/GPR motif that shares no homology with R6A-1 and binds only to Galpha(i1-3) in this assay. Binding of R6A-1 is generally specific to the GDP-bound state of the Galpha subunits and excludes association with Gbetagamma. R6A-Galpha(i1) complexes are resistant to trypsin digestion and exhibit distinct stability in the presence of Mg(2+), suggesting that the R6A and GPR peptides exert their activities using different mechanisms. Studies using Galpha(i1)/Galpha(s) chimeras identify two regions of Galpha(i1) (residues 1-35 and 57-88) as determinants for strong R6A-G(ialpha1) interaction. Residues flanking the R6A-1 peptide confer unique binding properties, indicating that the core motif could be used as a starting point for the development of peptides exhibiting novel activities and/or specificity for particular G protein subclasses or nucleotide-bound states.


Vitamin D status is associated with sociodemographic factors, lifestyle and metabolic health.
Low serum 25(OH)D concentration has been shown to predict the occurrence of several chronic diseases. It is, however, still unclear whether the associations are causal or due to confounding. The aim of this study was to investigate the associations between serum 25(OH)D concentration and sociodemographic, lifestyle and metabolic health-related factors. The study population comprised 5,714 men and women, aged 30-79 years, from the Health 2000 Survey representing the Finnish population. Serum 25(OH)D concentration was determined by radioimmunoassay from serum samples frozen at -70 °C. Sociodemographic, lifestyle and metabolic factors were determined by questionnaires, interviews and measurements. Linear regression was used to assess the associations between serum 25(OH)D and the factors studied. The mean serum 25(OH)D concentration was 45.3 nmol/l and it varied between categories of sociodemographic, lifestyle and metabolic health variables. Older age, being married or cohabiting and higher education were related to higher serum 25(OH)D concentration. Those with the healthiest lifestyle estimated by a lifestyle index based on body mass index, physical activity, smoking, alcohol consumption and diet had 15.8 nmol/l higher serum 25(OH)D concentration compared to those with the unhealthiest lifestyle. Of the indicators of metabolic health, only waist circumference and HDL cholesterol were significantly associated with 25(OH)D after adjustment for sociodemographic, lifestyle and other metabolic health factors. This study suggests that serum 25(OH)D concentration is associated with a multitude of sociodemographic, lifestyle and metabolic health factors. Thus, it is possible that such factors confound associations observed between serum 25(OH)D concentration and chronic diseases.


Lysine decarboxylase assay by the pH-stat method.
A simple assay of lysine decarboxylase is described. It is based on measuring the rate of titration of OH- ions, released during decarboxylation with the aid of a pH-stat apparatus. The continuous recording of the reaction progress may be easily performed. Using the pH-stat method the Km for lysine decarboxylase from E. coli is 2.0 mM.


Determination of isoflavones in rat serum using liquid chromatography-tandem mass spectrometry with a highly efficient core-shell column.
Consumption and nutritional supplementation of soy and soy-based products have been linked to health benefits such as lower cholesterol and triglyceride levels, and decreased incidence of cardiovascular disease and diabetes. In this study, we have developed a sensitive, specific, and robust method using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) for determination of serum isoflavones. A new highly efficient pentafluorophenyl phase core-shell column was first used to separate all isoflavones within 3 min, a separation time which is comparable to ultra-pressure liquid chromatography (UPLC) and micro-HPLC. A two-enzyme hydrolysis system with sulfatase and β-glucuronidase has also been developed to improve the efficiency of deconjugation of conjugated isoflavones in serum. The corresponding conjugated isoflavones were used to evaluate recoveries. In addition to duplicates, the method of standard addition was also applied in sample analysis for quality control. The developed method was applied to the analysis of 32 serum samples and was shown to be specific, sensitive and reproducible.


The influence of acute hypoxia and sleep states on the electrical activity of the cerebellum in the sheep fetus.
The experiments were designed to examine (1) the effects of acute hypoxia and (2) the influence of sleep states on the electrical activity of the cerebellum in the fetus. Five fetal sheep were instrumented between 106 and 133 days of gestation and survived in utero until term (140-146 days). Insulated stainless steel electrodes (75 microns diam.) were introduced into the cerebellar cortex and cemented in place. Electrodes were also implanted into 4 fetuses to monitor the electrocorticogram, rapid eye movements and the EMG of the dorsal neck (nuchal) muscles. Recordings of cerebellar activity were made for 252 h in 4 fetuses. The recordings were derived from many units (probably Purkinje cells) with a high level of basal discharge. The mean firing frequency was related to fetal sleep state. The highest frequencies coincided with episodes of rapid eye movements with low-voltage electrocortical activity and the lowest occurred during high-voltage electrocortical activity when rapid eye movements are absent. Acute hypoxaemia (fetal pa O2 6-10 mm Hg for up to 30 min) abolished rapid eye movements and caused a marked reduction in cerebellar activity. High-voltage electrocortical activity and nuchal EMG activity were maintained indicating that the effects of hypoxia on the cerebellum were not due simply to a general CNS depression. Cerebellar neural activity returned to normal at the termination of the hypoxic stimuli demonstrating that fetal Purkinje cells can withstand short episodes of severe hypoxia.


Picket-fences in the plasma membrane: functions in immune cells and phagocytosis.
Recent studies of molecular mobility in the plasma membrane have revealed that diffusion is restricted by cytoskeletal networks or fences. Transmembrane protein "pickets" that reversibly associate with the membrane-associated skeleton and with the pericellular coat impede the movement of unattached bystander molecules. While membrane picket-fences were originally described as barriers to free diffusion in more passive cell types such as fibroblasts, they have particularly important functions in the more dynamic immune cells. In phagocytes, such fences curtail spontaneous activation and their disassembly facilitates stimulation by target particles, fostering receptor clustering and the exclusion of phosphatases from the phagocytic cup. In this review, we describe the nature of the cellular cytoskeleton and of the exoskeleton created by the pericellular coat, their association with transmembrane pickets, and the modulation of molecular mobility during phagocytosis.


Nonmelanoma skin cancer.
Nonmelanoma skin cancers encompass a vast array of malignancies: adventitial, vascular, neuroendocrine, hematologic, adnexal, and epithelial. The skin is also a major site for metastasis of internal disease. With an understanding of the frequency of presentation, clinical distribution, and appearance, the clinician is able to provide an accurate diagnosis for most lesions. Because there are so many nonmelanoma skin cancers, the focus here is on those with a high frequency or significant morbidity. The treatment of nonmelanoma skin cancer is constantly evolving; therefore, emphasis is on diagnosis.


No Association Between Intrauterine Contraceptive Devices and Musculoskeletal Hip Joint Pain.
To investigate the association between intrauterine device (IUD) use and hip pain, orthopaedic visits for hip pain, and arthroscopic hip surgery. This was a retrospective cohort study of patients aged 18-44 years old using either IUDs or subdermal implants for contraception in a large commercial claims database (MarketScan) from 2012 to 2015. All patients had at least 12 months of continuous enrollment both before and after contraceptive placement. Patients with a history of hip pain or surgery were excluded. The primary outcome was new hip pain. Secondary outcomes included visiting an orthopaedic or sports medicine provider for a hip complaint, intra-articular hip injection, and arthroscopic hip surgery. Outcomes were analyzed with Cox proportional-hazard models. We identified a total of 242,383 patients, including 216,541 (89.3%) with IUDs and 25,842 (10.7%) with subdermal contraceptive implants. In time-to-event analysis, IUDs (vs implants) were not associated with increased risk of new hip pain diagnoses (hazard ratio [HR] 0.95, 95% confidence interval [CI] 0.87-1.03, P = .21). In contrast, both age (P < .001) and region (P < .001) were associated with increased risk of new hip pain. Similar results were seen for the secondary outcomes, including risk of orthopaedic visits for hip complaints (HR 1.06, 95% CI 0.83-1.35, P = .63), intra-articular injections of the hip (HR 0.94, 95% CI 0.63-1.41, P = .77), and hip arthroscopy procedures (HR 1.13, 95% CI 0.53-2.40, P = .75). In this study, we found no evidence that IUDs were associated with hip pain or surgery. Level III, retrospective cohort.


Physical therapy to improve functioning of older people in residential care facilities.
The purpose of this study was to determine the effectiveness of an individualized physical therapy mobility training program on the gait, balance, and functional performance of elderly individuals living in residential care facilities. Twenty-seven elderly individuals with impaired balance and difficulty performing at least one functional activity participated in the study. The subjects ranged in age from 71 to 97 years (mean = 87.1, SD = 6.7). Balance and gait speed were assessed at baseline and following physical therapy that consisted of exercises to improve specific functional limitations. Outcomes were reassessed 1 month following completion of the physical therapy. Gait and balance outcomes were analyzed using a one-way repeated-measures analysis of variance. Improvement was obtained in balance, which was maintained at 1 month follow-up. Gait speed did not improve to a level of statistical significance. After physical therapy, subjects improved in balance and functional performance. An improvement in gait speed may require a longer duration of treatment.


Spatial organization of EEGs from olfactory bulb and cortex.
This report presents evidence for the concept of 'wave packet' (Freeman 1975) in the olfactory system. EEG bursts recorded from electrodes chronically implanted at multiple sites in the olfactory bulb and cortex in awake rabbits were spatially coherent, iso-frequency events. Both bulbar and cortical bursts were composed of a major oscillation in the 35-85 c/sec range. The dominant cortical frequency was within 5% of that of the bulb. A secondary oscillation in the 15-35 c/sec range was found in both bulb and cortex. This was thought to represent a feedback signal of the cortex to the bulb.


Effect of mild chronic stress, as a model of depression, on the immunoreactivity of C57BL/6 mice.
Numerous studies correlate the state of depression with some abnormalities in the immune response, such as increased numbers of white blood-cells, alterations in sub-populations of leucocytes, suppression of cytotoxic activity of natural-killer cells, increased levels of some autoantibodies and acute-phase proteins. Some of these changes have been attributed to autoimmunological reactions. While the possibilities to evaluate some reactions in depressed patients are limited, an animal model of depression could well simulate this clinical situation, and the chronic mild state of stress is a well accepted one. After undergoing stress for three-weeks, C57BL/6 mice demonstrate in the present study a decrease in thymus weight, as well as increased interleukin-1 and decreased interleukin-2 production. Splenocytes of the depressed mice exert a decrease in natural-killer-cell activity, in the proliferative response to Concanavalin-A, interleukin-1 and anti-CD3 monoclonal antibodies and an increase in the proliferative response to lipopolysaccharides and pokeweed mitogens. Our results also suggest that chronic stress-induced activation of suppressor cells in the spleen, due to elimination of CD8+ cells, increase the proliferation of splenocytes in response to mitogens of T cells.


Prosthodontic rehabilitation of a patient with nintedanib-induced osteonecrosis of the jaw: A clinical report.
Medication-related osteonecrosis of the jaw results in extensive excision and proves challenging to rehabilitate prosthetically. This article describes the prosthodontic rehabilitation of medication-related osteonecrosis of the jaw induced by nintedanib. A custom-made subperiosteal implant was designed to adapt to the bone, remaining after the excision of the jaw necrosis. The implant followed the anatomic contour of the residual bone and was milled in titanium alloy, allowing the rehabilitation of a patient with a fixed dental prosthesis.


Thuricin: the bacteriocin produced by Bacillus thuringiensis.
Bacillus thuringiensis serovar, thuringiensis (HD-2) demonstrated antibacterial activity against 48 of 56 strains of B. thuringiensis and against some other Gram-positive species but not against Gram-negative species. The antibacterial activity was not inducible by mitomycin C or by ultraviolet irradiation, and additional activity was not liberated from cells by sonication. Upon dilution of the antibacterial substance, zones of inhibition diminished without the appearance of plaques. Gel filtration chromatography indicated an Mr greater than 950,000 for the bacteriocin (thuricin) in its native form. The native thuricin was sedimented by ultracentrifugation, but electron microscopy of the pellet failed to reveal phage particles or phage components. Nondenaturing polyacrylamide gel electrophoresis (PAGE) of thuricin demonstrated the association of bacteriocin activity with a protein band which migrated only slightly into a 5% gel. Sodium dodecyl sulfate (SDS)-PAGE of partially purified thuricin revealed five major bands. Thuricin activity was substantially reduced by treatment with chymotrypsin, pronase, subtilisin, trypsin, and heat at 96 degrees C but not by treatment with lysozyme, phospholipase C, papain, peptidase, or organic solvents. It exhibited a bactericidal and bacteriolytic effect on a sensitive strain, B. thuringiensis serovar, canadensis (MF4). Partially purified preparations of thuricin had phospholipase A activity which was adsorbed by sensitive cells but not by cells which were insensitive to thuricin. Antibacterial activity was blocked by preincubation of thuricin with phospholipid. Loss of a 150-mDa plasmid was correlated with loss of thuricin production.


Mycotic Aneurysm of the Common Carotid Artery as a Presenting Symptom for Early Colorectal Malignancy.
Mycotic carotid aneurysms are rare vascular conditions that are associated with high mortality if left untreated. We present a gentleman who had a progressively enlarging tender pulsatile swelling on the right side of neck. Emergency computed tomography scan showed a large mycotic aneurysm arising from the right common carotid artery, and he underwent emergency open exploration with interposition bypass graft. Bacteroides fragilis species, which signified gastrointestinal-related sepsis, was isolated from the thrombus culture. Subsequently search of systemic septic source showed an early rectal adenocarcinoma. This is the first case in the world's literature of a patient who had B. fragilis mycotic carotid aneurysm as a presenting complaint of his occult rectal malignancy.


Transporting the Burden of Justification: The Unethicality of Transgender Conversion Practices.
Transgender conversion practices involve attempts to alter, discourage, or suppress a person's gender identity and/or desired gender presentation, including by delaying or preventing gender transition. Proponents of the practices have argued that they should be allowed until proven to be harmful. Drawing on the notion of expressive equality, I argue that conversion practices are prima facie unethical because they do not fulfill a legitimate clinical purpose and conflict with the self-understanding of trans communities.


Acoustic resolution photoacoustic microscopy based on microelectromechanical systems scanner.
Photoacoustic microscopy (PAM) can be classified as optical resolution (OR)-PAM and acoustic resolution (AR)-PAM depending on the type of resolution achieved. Using microelectromechanical systems (MEMS) scanner, high-speed OR-PAM system was developed earlier. Depth of imaging limits the use of OR-PAM technology for many preclinical and clinical imaging applications. Here, we demonstrate the use of a high-speed MEMS scanner for AR-PAM imaging. Lateral resolution of 84 μm and an axial resolution of 27 μm with ~2.7 mm imaging depth was achieved using a 50 MHz transducer-based AR-PAM system. Use of a higher frequency transducer at 75 MHz has further improved the resolution characteristics of the system with a reduction in imaging depth and a lateral resolution of 53 μm and an axial resolution of 18 μm with ~1.8 mm imaging depth was achieved. Using the two-axis MEMS scanner a 2 × 2 .5 mm2 area was imaged in 3 seconds. The capability of achieving acoustic resolution images using the MEMS scanner makes it beneficial for the development of high-speed miniaturized systems for deeper tissue imaging.


Effect of aminoglutethimide on urinary cortisol and cortisol metabolites in adolescents with Cushing's syndrome.
The effect of aminoglutethimide (AG) 4 X 250 mg (670 mg/m2 daily by mouth) on the excretion of the free cortisol (radio-immunoassay) and of its metabolites THE, THF-allo THF, cortolone and beta-cortolone (gas chromatography on capillary column) was studied monthly during 3-5 months in four adolescents (one girl, three boys) aged 15.9-18 years with Cushing's syndrome due to bilateral adrenal hyperplasia, but without evidence of a pituitary tumour. Under AG, all compounds decreased to a minimum after 1-2 months. The decrease of THE- THF-allo THF was most marked, followed by cortolone-beta-cortolone and free cortisol. The sum of the conjugated metabolites was normalized, but free cortisol remained high. A rebound was noted after 3-5 months of continued treatment. This was associated with clinical relapse (weight gain, increasing blood pressure). With AG, a non-steroidal peak appeared on the chromatograms. It is concluded that: (1) AG is only temporarily effective in diminishing the excretion of cortisol and its metabolites; (2) paradoxical increments of 17-ketosteroids as reported from colorimetric analysis are non-specific and are probably due to the non-steroidal peak; and (3) AG appears to modify steroid catabolizing liver enzymes (inhibition of 5beta-reductase and/or 3alpha-dehydrogenase, possibly stimulation of 20alpha- and 20beta-dehydrogenases). This could increase the biological half-life of cortisol and contribute to the clinical rebound, which is due to increased ACTH-secretion. Because of its excellent short-term effects, AG appears to be useful to prepare patients for bilateral adrenalectomy.


The effect of diet on intermediary metabolite concentrations in Type 2 (non-insulin-dependent) diabetes mellitus.
The metabolic response to a standard meal was studied in six Type 2 (non-insulin-dependent) diabetic patients at diagnosis and following 4-6 weeks of dietary treatment. The fall in blood glucose concentration following treatment was accompanied by significant reductions in circulating concentrations of lactate, pyruvate, alanine and glycerol. Blood 3-hydroxybutyrate concentrations also fell with treatment.


The conformation of glycans of the oligo-D-mannosidic type, and their interaction with concanavalin A: a computer-modelling study.
The favored conformations of glycans of the oligo-D-mannosidic type have been determined by using empirical energy calculations. An interesting aspect is that the alpha-(1----3)-linked terminal D-mannose residue of the outer trimannosidic core fragment, in all the conformations which fall within 5 kcal.mol-1 of the global minimum, always lies close to the chitobiose core. These models are in general agreement with the available n.m.r. data. The probable modes of binding of these glycans to concanavalin A (Con A) were determined, by using a computer-modelling technique which identifies the positions for the different conformers of the carbohydrate in the binding site of Con A, based on stereochemical considerations. These studies showed that Con A can bind only to two of the three terminal D-mannose residues in these glycans, because the D-mannose residue which lies close to the chitobiose core is inaccessible for the binding of Con A. Of these two terminal D-mannose residues, the alpha-(1----6)-linked D-mannose may bind the more strongly. Furthermore, it is shown that the internal D-mannose residue will, at best, interact very weakly with the carbohydrate-binding site of Con A. These results rationalize well the available data on the binding affinity of these glycans to Con A. They further support the conclusion that the binding affinity of a glycan to Con A does not depend on the number in the glycan, of D-mannose residues which possess free 3-, 4-, and 6-hydroxyl groups, but, rather, on the accessibility of these residues to Con A.


Assessment of drug salt release from solutions, suspensions and in situ suspensions using a rotating dialysis cell.
A rotating dialysis cell consisting of a small (10 ml) and a large compartment (1000 ml) was used to study the release of drug salt (bupivacaine 9-anthracene carboxylate) from (i). solutions, (ii). suspensions and (iii). in situ formed suspensions. Initial release experiments from suspensions indicated that the release of drug salt in deionized water was predominantly limited by the diffusion across the membrane whereas it is essentially dissolution rate controlled in 0.05 M phosphate buffer (pH 7.40). Thus, the in vitro model appears to have a potential in formulation screening when phosphate buffer is used as release media. Generally, the initial release of the drug salt from in situ suspensions occurred faster as compared to conventional suspensions, probably due to incomplete precipitation of the drug salt, and hence formation of supersaturated solutions where the rate of release is predominantly determined by the concentration gradient. However, when an adequately concentrated solution of the drug salt was used to prepare the in situ suspension, the initial fast release was followed by a substantial sustained release indicating that the release had become dissolution rate limited.


Two unusual cases of pharyngeal hairy polyp causing intermittent neonatal airway obstruction.
Hairy polyp of the pharynx is an uncommon developmental malformation that is most frequently seen as a penduculated tumour in the neonate. The clinical presentation is characterized by the presence of a polypoid mass protruding through the mouth as 'a second tongue' causing respiratory distress. Two patients are presented with this condition.


Efficient preparation of plant chromosomes for high-resolution scanning electron microscopy.
A highly reproducible technique to prepare plant chromosomes for high-resolution field emission scanning electron microscopy is presented. The procedure allows the production of relatively high numbers of chromosome spreads that can be viewed at high resolution, showing structural details below 10 nm. This preparation technique is not restricted to metaphase chromosomes, but also allows the observation of plant chromosomes during all stages of the cell cycle.


Fitness and competitive growth comparison of methicillin resistant and methicillin susceptible Staphylococcus aureus colonies.
Exponential developments of both Methicillin resistant Staphylococcus aureus (MRSA) 3R ve 36R and methicillin susceptible Staphylococcus aureus (MSSA) 27S were evaluated in the presence and absence of oxacillin. The strains were isolated from the specimens collected in microbiology department. It was also determined the transfer of mecA gene from 3R to 27S strain by using the replica plate technique. It was observed that the presence of antibiotics in the preliminary culture had a positive impact on the growth of the secondary culture of MRSA isolates. Comparison results of Rt bacteria in three different mixed cultures, assessed with Tukey's HSD test, showed a significant statistical difference among the groups. The values were as following; on the first day; Df: 2, F: 60.90, P: 0.0001, second day; Df:2, F:90.56, P: 0.0000, and third day; Df:2, F:4.86, P:0.0557. As a result of the study, we can suggest that the gene expression levels of the transferred antibiotic resistance genes could help us in both controlling hospital originated sickness and developing new strategies to prevent the spread of resistant bacteria.


Baicalin induces apoptosis via mitochondrial pathway as prooxidant.
Baicalin is a flavonoid and a major component of a herbal medicine, Sho-saiko-to, which is commonly used for treatment of chronic hepatitis in Japan and China. Flavonoids including baicalin have been reported to not only function as anti-oxidants but also cause cytotoxic effect. We investigated the mechanism of baicalin-induced cytotoxicity in leukemia-derived T cell line, Jurkat cells. When cells were cultured with 50-200 microg/ml baicalin for 6h, caspase-3 was activated and then cells fell into apoptosis. Induction of apoptosis by baicalin was accompanied with the marginal generation of intracellular reactive oxygen species (ROS), the increase of the cytosolic fractions of cytochrome c, and the disruption of mitochondrial transmembrane potential (DeltaPsi(m)) prior to the activation of caspase-3. The pre-culture with 5 mM of buthionine sulfoximine (BSO), an inhibitor of glutathione (GSH) synthesis, facilitated baicalin-induced disruption of DeltaPsi(m) and induction of apoptosis. The pre-culture with N-benzyloxycarbonyl-valyl-alanyl-aspartyl fluoromethylketone (Z-VAD-fmk), a pan-caspase inhibitor, partially suppressed the induction of apoptosis. On the other hand, baicalin showed little toxic effect on peripheral blood mononuclear cells (PBMCs) from healthy volunteers. These results indicate that baicalin acts as a prooxidant and induces caspase-3 activation and apoptosis via mitochondrial pathway.


Nontraumatic subdural hematoma in a patient with osteogenesis imperfecta and renal failure.
Intracranial hematomas in patients with osteogenesis imperfecta rarely are reported despite an increased incidence of skull fractures. We present a case of a patient with osteogenesis imperfecta but without any history of trauma or chronic anticoagulation in whom a subdural hematoma occurred during hemodialysis. Despite operative intervention, the patient died. We could find no other cases in the medical literature of a patient with osteogenesis imperfecta who had an intracranial hematoma in the absence of a skull fracture. Patients with osteogenesis imperfecta may be at high risk for intracranial bleeding from coagulation abnormalities or otherwise trivial injuries.


A clinical and microbiological study of Actinomycetes in oral and cervicofacial lesions.
A retrospective study of the incidence of cervicofacial actinomycosis in patients attending the Liverpool Dental Hospital from 1980-1983 was undertaken. A total of 86 patients were found which were divisable into three distinct presentations: acute painful swellings with a duration of less than 1 month; chronic long-standing infections, duration greater than 3 months; unsuspected microbiologically-proven actinomycotic lesions. A prospective study from 1983-1986 also was divisable into three similar types of presentations. The most common presentation in both parts of the study was acute painful swellings associated with soft tissue abscesses secondary to dental lesions. Clinicians dealing with acute dentally-associated swellings should consider the possibility of actinomycosis as a possible diagnosis.


Metabolomics of papillary thyroid carcinoma tissues: potential biomarkers for diagnosis and promising targets for therapy.
Papillary thyroid carcinoma (PTC) is the most common pathological type of thyroid cancer. Our study was to construct a tissue-targeted metabolomics analysis method based on untargeted and targeted metabolic multi-platforms to identify a comprehensive PTC metabolic network in clinical samples. We applied untargeted gas chromatography-time-of-flight mass spectrometry (GC-TOF-MS) for preliminary screening of potential biomarkers. With diagnostic models constructed using principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA) and orthogonal partial least squares discriminant analysis (OPLS-DA), 45 differentially abundant metabolites with a variable importance in the projection (VIP) value greater than 1 and a P value less than 0.05 were identified, and we show that our approach was able to discriminate PTC tissues from healthy tissues. We then performed validation experiments based on targeted GC-TOF-MS combined with ultra-high-performance liquid chromatography-triple-quadrupole mass spectrometry (UHPLC-QqQ-MS) through constructing linear standard curves of analytes. Ultimately, galactinol, melibiose, and melatonin were validated as significantly altered metabolites (p < 0.05). These three metabolites were defined as a combinatorial biomarker to assist needle biopsy for PTC diagnosis as demonstrated by receiver operating characteristic (ROC) curve analysis, which revealed an area under the ROC curve (AUC) value of 0.96. Based on the metabolite enrichment analysis results, the galactose metabolism pathway was regarded as an important factor influencing PTC development by affecting energy metabolism. Alpha-galactosidase (GLA) was considered to be a potential target for PTC therapy.


One-stage excision of massive cervicomediastinal lymphangioma in the newborn.
We report the first neonate successfully treated with one-stage complete excision of massive cervicomediastinal lymphangioma producing alarming respiratory distress. Early postoperative course was complicated by refractory bilateral chylothorax, which effectively responded to insertion of external pleuroperitoneal shunts. At the 6-year follow-up, the child remains symptom free and recurrence free. One-stage resection for cervicomediastinal lymphangioma is curative and well tolerated even shortly after birth.


Thallium-201 Imaging in Intact Olfactory Sensory Neurons with Reduced Pre-Synaptic Inhibition In Vivo.
In this study, we determined whether the 201Tl (thallium-201)-based olfactory imaging is affected if olfactory sensory neurons received reduced pre-synaptic inhibition signals from dopaminergic interneurons in the olfactory bulb in vivo. The thallium-201 migration rate to the olfactory bulb and the number of action potentials of olfactory sensory neurons were assessed 3 h following left side nasal administration of rotenone, a mitochondrial respiratory chain complex I inhibitor that decreases the number of dopaminergic interneurons without damaging the olfactory sensory neurons in the olfactory bulb, in mice (6-7 animals per group). The migration rate of thallium-201 to the olfactory bulb was significantly increased following intranasal administration of thallium-201 and rotenone (10 μg rotenone, p = 0.0012; 20 μg rotenone, p = 0.0012), compared with that in control mice. The number of action potentials was significantly reduced in the olfactory sensory neurons in the rotenone treated side of 20 μg rotenone-treated mice, compared with that in control mice (p = 0.0029). The migration rate of thallium-201 to the olfactory bulb assessed with SPECT-CT was significantly increased in rats 24 h after the left intranasal administration of thallium-201 and 100 μg rotenone, compared with that in control rats (p = 0.008, 5 rats per group). Our results suggest that thallium-201 migration to the olfactory bulb is increased in intact olfactory sensory neurons with reduced pre-synaptic inhibition from dopaminergic interneurons in olfactory bulb glomeruli.