Predicting skin permeability from complex chemical mixtures.
Occupational and environmental exposure to topical chemicals is usually in the form of complex chemical mixtures, yet risk assessment is based on experimentally derived data from individual chemical exposures from a single, usually aqueous vehicle, or from computed physiochemical properties. We present an approach using hybrid quantitative structure permeation relationships (QSPeR) models where absorption through porcine skin flow-through diffusion cells is well predicted using a QSPeR model describing the individual penetrants, coupled with a mixture factor (MF) that accounts for physicochemical properties of the vehicle/mixture components. The baseline equation is log k(p) = c + mMF + a sigma alpha2(H) + b sigma beta2(H) + s pi2(H) + rR2 + vV(x) where sigma alpha2(H) is the hydrogen-bond donor acidity, sigma beta2(H) is the hydrogen-bond acceptor basicity, pi2(H) is the dipolarity/polarizability, R2 represents the excess molar refractivity, and V(x) is the McGowan volume of the penetrants of interest; c, m, a, b, s, r, and v are strength coefficients coupling these descriptors to skin permeability (k(p)) of 12 penetrants (atrazine, chlorpyrifos, ethylparathion, fenthion, methylparathion, nonylphenol, rho-nitrophenol, pentachlorophenol, phenol, propazine, simazine, and triazine) in 24 mixtures. Mixtures consisted of full factorial combinations of vehicles (water, ethanol, propylene glycol) and additives (sodium lauryl sulfate, methyl nicotinate). An additional set of 4 penetrants (DEET, SDS, permethrin, ricinoleic acid) in different mixtures were included to assess applicability of this approach. This resulted in a dataset of 16 compounds administered in 344 treatment combinations. Across all exposures with no MF, R2 for absorption was 0.62. With the MF, correlations increased up to 0.78. Parameters correlated to the MF include refractive index, polarizability and log (1/Henry's Law Constant) of the mixture components. These factors should not be considered final as the focus of these studies was solely to determine if knowledge of the physical properties of a mixture would improve predicting skin permeability. Inclusion of multiple mixture factors should further improve predictability. The importance of these findings is that there is an approach whereby the effects of a mixture on dermal absorption of a penetrant of interest can be quantitated in a standard QSPeR model if physicochemical properties of the mixture are also incorporated.


Ischemic chiasmal syndrome associated with posterior communicating artery (PCoA) and tuberothalamic artery (TA) infarction: a case report.
Lesions affecting the body of the optic chiasm typically produce bitemporal hemianopia. The blood supply comes from the anterior communicating artery, anterior cerebral, posterior communicating, posterior cerebral, and basilar arteries. We herein report a young patient admitted to the emergency department with acute confusion, left-sided hemiparesis, hemihypoesthesia, and dysarthria. Bitemporal hemianopia was detected after resolution of confusion. On cranial magnetic resonance imaging (MRI), infarction in the right anterolateral thalamus in the territory of tuberothalamic artery (TA) and in posterior chiasma in the territory of the posterior communicating artery (PCoA) was revealed. Cerebral MR angiography showed luminal irregularity of the PCoA. The patient was presented to draw attention to the rare entity ischemic chiasmal syndrome.


Concepts in Neural Stimulation: Electrical and Optical Modulation of the Auditory Pathways.
Understanding the mechanisms of neural stimulation is necessary to improve the management of sensory disorders. Neurons can be artificially stimulated using electrical current, or with newer stimulation modalities, including optogenetics. Electrical stimulation forms the basis for all neuroprosthetic devices that are used clinically. Off-target stimulation and poor implant performance remain concerns for patients with electrically based neuroprosthetic devices. Optogenetic techniques may improve cranial nerve stimulation strategies used by various neuroprostheses and result in better patient outcomes. This article reviews the fundamentals of neural stimulation and provides an overview of recent major advancements in light-based neuromodulation."


Early changes of fecal short-chain fatty acid levels in patients with mild cognitive impairments.
To compare the fecal levels of short-chain fatty acids (SCFAs) in patients with mild cognitive impairment (MCI) and normal controls (NCs) and to examine whether fecal SCFAs could be used as the biomarker for the identification of patients with MCI. To examine the relationship between fecal SCFAs and amyloid-β (Aβ) deposition in the brain. A cohort of 32 MCI patients, 23 Parkinson's disease (PD) patients, and 27 NC were recruited in our study. Fecal levels of SCFAs were measured using chromatography and mass spectrometry. Disease duration, ApoE genotype, body mass index, constipation, and diabetes were evaluated. To assess cognitive impairment, we used the Mini-Mental Status Examination (MMSE). To assess brain atrophy, the degree of medial temporal atrophy (MTA score, Grade 0-4) was measured by structural MRI. Aβ positron emission tomography with 18 F-florbetapir (FBP) was performed in seven MCI patients at the time of stool sampling and in 28 MCI patients at an average of 12.3 ± 0.4 months from the time of stool sampling to detect and quantify Aβ deposition in the brain. Compared with NC, MCI patients had significantly lower fecal levels of acetic acid, butyric acid, and caproic acid. Among fecal SCFAs, acetic acid performed the best in discriminating MCI from NC, achieved an AUC of 0.752 (p = 0.001, 95% CI: 0.628-0.876), specificity of 66.7%, and sensitivity of 75%. By combining fecal levels of acetic acid, butyric acid, and caproic acid, the diagnostic specificity was significantly improved, reaching 88.9%. To better verify the diagnostic performance of SCFAs, we randomly assigned 60% of participants into training dataset and 40% into testing dataset. Only acetic acid showed significantly difference between these two groups in the training dataset. Based on the fecal levels of acetic acid, we achieved the ROC curve. Next, the ROC curve was evaluated in the independent test data and 61.5% (8 in 13) of patients with MCI, and 72.7% (8 in 11) of NC could be identified correctly. Subgroup analysis showed that reduced fecal SCFAs in MCI group were negatively associated with Aβ deposition in cognition-related brain regions. Reductions in fecal SCFAs were observed in patients with MCI compared with NC. Reduced fecal SCFAs were negatively associated with Aβ deposition in cognition-related brain regions in MCI group. Our findings suggest that gut metabolite SCFAs have the potential to serve as early diagnostic biomarkers for distinguishing patients with MCI from NC and could serve as potential targets for preventing AD.


Demographic, Healthcare Access, and Dietary Factors Associated With Thyroid Hormone Treatments for Hypothyroidism.
Clinical guidelines have recommended a trial of liothyronine (LT3) with levothyroxine (LT4) in select patients with hypothyroidism. However, little is known about the real-world use of LT3 and desiccated thyroid extract (DTE) and the characteristics of patients treated with LT3 and DTE. (1) Determine national trends of new LT4, LT3, and DTE prescriptions in the United States; (2) determine whether sociodemographic, healthcare access, and dietary factors are associated with different thyroid hormone (TH) therapies. Parallel cross-sectional studies were conducted using 2 datasets: (1) a national patient claims dataset (2010-2020) and (2) the National Health and Nutrition Examination Study (NHANES) dataset (1999-2016). Included participants had a diagnosis of primary or subclinical hypothyroidism. Study outcomes included the impact of demographics and healthcare access on differences in the proportion of TH therapies consisting of LT4, LT3, and DTE (patient claims) and differences in dietary behaviors between DTE-treated participants and LT4-treated matched controls (NHANES). On an average annual basis, 47 711 adults received at least 1 new TH prescription, with 88.3% receiving LT4 monotherapy, 2.0% receiving LT3 therapy, and 9.4% receiving DTE therapy. The proportion receiving DTE therapy increased from 5.4% in 2010 to 10.2% in 2020. In the analysis between states, high primary care and endocrinology physician densities were associated with increased use of LT4 monotherapy (odds ratio 2.51, P < .001 and odds ratio 2.71, P < .001). DTE-treated NHANES participants (n = 73) consumed more dietary supplements compared to LT4-treated participants (n = 146) (4.7 vs 2.1, P < .001). The proportion of new TH therapies containing DTE for hypothyroidism doubled since 2010 while LT3 therapies remained stable. DTE treatment was associated with decreased physician density and increased dietary supplement use.


Genotoxic, teratotoxic and oral toxic assessments of Antrodia cinnamomea health food product (Leader Deluxe Antrodia cinnamomea®).
Antrodia cinnamomea is a rare and endemic medicinal mushroom native to Taiwan. The pharmacological effects of A. cinnamomea have been extensively studied. The aim of the present study was to assess the genotoxic, oral toxic and teratotoxic effects of A. cinnamomea health food product ⿿Leader Deluxe Antrodia cinnamomea (LDAC)⿿⿿ using in vitro and in vivo tests. The Ames test with 5 strains of Salmonella typhimurium showed no signs of increased reverse mutation upon exposure to LDAC up to concentration of 5 mg/plate. Exposure of Chinese Hamster Ovary cells (CHO-K1) to LDAC did not produce an increase in the frequency of chromosomal aberration in vitro. In addition, LDAC treatment did not affect the proportions of immature to total erythrocytes and the number of micronuclei in the immature erythrocytes of ICR mice. Moreover, 14-days single-dose acute toxicity and 90-days repeated oral dose toxicity tests with rats showed that no observable adverse effects were found. Furthermore, after treatment with LDAC (700⿿2800 mg/kg/day) there was no evidence of observable segment II reproductive and developmental toxic effects in pregnant SD rats and their fetuses. These toxicological assessments support the safety of LDAC for human consumption.


Different Heat Shock Proteins Bind α-Synuclein With Distinct Mechanisms and Synergistically Prevent Its Amyloid Aggregation.
α-Synuclein (α-Syn) forms pathological amyloid aggregates deposited in Lewy bodies and Lewy neurites in the brain of Parkinson's disease (PD) patients. Heat shock proteins (Hsps) are the major components of the cellular chaperone network, which are responsible for preventing proteins from amyloid aggregation. Different Hsps were reported to interact with α-syn. However, the underlying mechanism of the interplay between α-syn and different Hsps remains unclear. Here, by combing NMR spectroscopy, electron microscope and other biochemical approaches, we systemically investigated the interaction between α-syn and three Hsps from different families including Hsp27, HDJ1, and Hsp104. We found that all three Hsps can weakly bind to α-syn and inhibit it from amyloid aggregation. Intriguingly, different Hsps recognize distinct regions of α-syn monomer, and act synergistically in chaperoning α-syn from fibril formation in sub-stoichiometry. Our results revealed the diverse binding mechanisms employed by different Hsps to tackle α-syn, and suggested that different Hsps form a network for cooperatively chaperoning α-syn from pathological aggregation.


HIV-1 hypermethylated guanosine cap licenses specialized translation unaffected by mTOR.
Appended to the 5' end of nascent RNA polymerase II transcripts is 7-methyl guanosine (m7G-cap) that engages nuclear cap-binding complex (CBC) to facilitate messenger RNA (mRNA) maturation. Mature mRNAs exchange CBC for eIF4E, the rate-limiting translation factor that is controlled through mTOR. Experiments in immune cells have now documented HIV-1 incompletely processed transcripts exhibited hypermethylated m7G-cap and that the down-regulation of the trimethylguanosine synthetase-1-reduced HIV-1 infectivity and virion protein synthesis by several orders of magnitude. HIV-1 cap hypermethylation required nuclear RNA helicase A (RHA)/DHX9 interaction with the shape of the 5' untranslated region (UTR) primer binding site (PBS) segment. Down-regulation of RHA or the anomalous shape of the PBS segment abrogated hypermethylated caps and derepressed eIF4E binding for virion protein translation during global down-regulation of host translation. mTOR inhibition was detrimental to HIV-1 proliferation and attenuated Tat, Rev, and Nef synthesis. This study identified mutually exclusive translation pathways and the calibration of virion structural/accessory protein synthesis with de novo synthesis of the viral regulatory proteins. The hypermethylation of select, viral mRNA resulted in CBC exchange to heterodimeric CBP80/NCBP3 that expanded the functional capacity of HIV-1 in immune cells.


Endocrinologic Control of Men's Sexual Desire and Arousal/Erection.
Several hormones and neurotransmitters orchestrate men's sexual response, including the appetitive (sexual desire) and consummative (arousal and penile erection) phases. To provide an overview and recommendations regarding endocrinologic control of sexual desire and arousal and erection and their disturbances. Medical literature was reviewed by the subcommittee of the International Consultation of Sexual Medicine, followed by extensive internal discussion, and then public presentation and discussion with other experts. The role of pituitary (prolactin, oxytocin, growth hormone, and α-melanocyte-stimulating hormone), thyroid, and testicular hormones was scrutinized and discussed. Recommendations were based on grading of evidence-based medical literature, followed by interactive discussion. Testosterone has a primary role in controlling and synchronizing male sexual desire and arousal, acting at multiple levels. Accordingly, meta-analysis indicates that testosterone therapy for hypogonadal individuals can improve low desire and erectile dysfunction. Hyperprolactinemia is associated with low desire that can be successfully corrected by appropriate treatments. Oxytocin and α-melanocyte-stimulating hormone are important in eliciting sexual arousal; however, use of these peptides, or their analogs, for stimulating sexual arousal is still under investigation. Evaluation and treatment of other endocrine disorders are suggested only in selected cases. Endocrine abnormalities are common in patients with sexual dysfunction. Their identification and treatment is strongly encouraged in disturbances of sexual desire and arousal.


Multiple fluorescence quenching effects mediated fluorescent sensing of captopril Based on amino Acids-Derivative carbon nanodots.
Carbon nanodots (CNDs) were facilely synthesized through a pyrolysis procedure with histamine, an amino acid rich in element carbon and nitrogen, being the precursor. Taking advantage of the favorable fluorescence performance of CNDs, a multiple fluorescence quenching effects mediated fluorescent sensor was established for captopril (CAP) detection. MnO2 NPs were firstly combined with CNDs via electrostatic attraction and subsequently quenched the fluorescence. The quenching mechanisms were concluded to be the combined effects of fluorescence resonance energy transfer (FRET) and inner filtration effect (IFE). Subsequently CAP triggered a unique redox reaction and decomposed the quencher so that renewed the fluorescence. Hence, the sensitive and selective detection of CAP was achieved through the indication of fluorescence recovery efficiency. A proportional range of 0.4 ∼ 60 μmol L-1 with the LOD of 0.31 μmol L-1 was obtained. The sensor was further applied to the real sample detection and the satisfactory results revealed the practical value of CNDs. The facile synthesis of CNDs and brand-new sensing mechanism made it a novel fluorescent method and could improve the analysis of CAP.


Cross-dehydrogenative coupling and oxidative-amination reactions of ethers and alcohols with aromatics and heteroaromatics.
Cross-dehydrogenative coupling (CDC) is a process in which, typically, a C-C bond is formed at the expense of two C-H bonds, either catalyzed by metals or other organic compounds, or via uncatalyzed processes. In this perspective, we present various modes of C-H bond-activation at sp3 centers adjacent to ether oxygen atoms, followed by C-C bond formation with aromatic systems as well as with heteroaromatic systems. C-N bond-formation with NH-containing heteroaromatics, leading to hemiaminal ethers, is also an event that can occur analogously to C-C bond formation, but at the expense of C-H and N-H bonds. A large variety of hemiaminal ether-forming reactions have recently appeared in the literature and this perspective also includes this complementary chemistry. In addition, the participation of C-H bonds in alcohols in such processes is also described. Facile access to a wide range of compounds can be attained through these processes, rendering such reactions useful for synthetic applications via Csp3 bond activations.


Morphological and transcriptional responses of Lycopersicon esculentum to hexavalent chromium in agricultural soil.
The carcinogenic, teratogenic, and mutagenic effects of hexavalent chromium (Cr[VI]) on living organisms through the food chain raise the immediate need to assess the potential toxicological impacts of Cr(VI) on human health. Therefore, the concentration-dependent responses of 12 Cr(VI)-responsive genes selected from a high-throughput Lycopersicon esculentum complementary DNA microarray were examined at different Cr concentrations. The results indicated that most of the genes were differentially expressed from 0.1 mg Cr/kg soil, whereas the lowest-observable-adverse-effect concentrations of Cr(VI) were 1.6 mg Cr/kg soil, 6.4 mg Cr/kg soil, 3.2 mg Cr/kg soil, and 0.4 mg Cr/kg soil for seed germination, root elongation, root biomass, and root morphology, respectively, implying that the transcriptional method was more sensitive than the traditional method in detecting Cr(VI) toxicity. Dose-dependent responses were observed for the relative expression of expansin (p = 0.778), probable chalcone-flavonone isomerase 3 (p = -0.496), and 12S seed storage protein CRD (p = -0.614); therefore, the authors propose the 3 genes as putative biomarkers in Cr(VI)-contaminated soil. Environ Toxicol Chem 2016;35:1751-1758. © 2015 SETAC.


The Justus-Warren Heart Disease and Stroke Prevention Task Force and the North Carolina plan to prevent heart disease and stroke.
One in 4 deaths in North Carolina is attributed to heart disease and stroke. The legislatively mandated Justus-Warren Heart Disease and Stroke Prevention Task Force provides statewide leadership for the prevention of cardiovascular disease. This commentary reviews the work and accomplishments of the task force and its comprehensive plan to reduce the state's cardiovascular burden.


Bone regeneration in calvarial defects in a rat model by implantation of human bone marrow-derived mesenchymal stromal cell spheroids.
Mesenchymal stem cell (MSC) condensation contributes to membrane ossification by enhancing their osteodifferentiation. We investigated bone regeneration in rats using the human bone marrow-derived MSC-spheroids prepared by rotation culture, without synthetic or exogenous biomaterials. Bilateral calvarial defects (8 mm) were created in nude male rats; the left-sided defects were implanted with MSC-spheroids, β-tricalcium phosphate (β-TCP) granules, or β-TCP granules + MSC-spheroids, while the right-sided defects served as internal controls. Micro-computed tomography and immunohistochemical staining for osteocalcin/osteopontin indicated formation of new, full-thickness bones at the implantation sites, but not at the control sites in the MSC-spheroid group. Raman spectroscopy revealed similarity in the spectral properties of the repaired bone and native calvarial bone. Mechanical performance of the bones in the MSC-implanted group was good (50 and 60% those of native bones, respectively). All tests showed poor bone regeneration in the β-TCP and β-TCP + MSC-spheroid groups. Thus, significant bone regeneration was achieved with MSC-spheroid implantation into bone defects, justifying further investigation.


Carbon dioxide laser transection of rat peripheral nerves.
Histologic, ultrastructural, and horseradish peroxidase tracing studies have shown that the carbon dioxide laser does not, as previously reported, seal transected axons in peripheral nerves that are subsequently repaired. If sealed nerve ends are the desired effect, such as in treatment of painful neuromas, this experimental animal study in rats suggests that the laser may be contraindicated because of the possible retention of proximal nerve regenerative ability. For resections of peripheral neuromas followed by neurorrhaphy, our data indicate that conventional scissor transection of the nerve may be followed by better reinnervation than that of laser transection.


Acceptability of Human Papilloma Virus Self-Sampling for Cervical Cancer Screening in a Cohort of Patients from Romania (Stage 2).
(1) Background: Low patient's adherence to conventional cervical cancer screening methods determined the need to take into consideration alternative approaches, and vaginal HPV self-sampling is one of them. We aimed to evaluate, using an online survey, the Romanian women's acceptability of vaginal HPV self-sampling. (2) Methods: A 13-questions online survey was distributed on three Facebook groups, and the results were summarized. (3) Results: Despite of good educational background, 10.8% (n = 60) of the respondents did not know what a Pap smear is, and 33% (n = 183) were not informed about the free national cervical cancer screening program. Multivariate analysis revealed an increased likelihood of vaginal self-sampling acceptance among respondents who did not know about Pap test (OR: 7.80; 95%CI: 1.062−57.431; p = 0.021), national cervical cancer screening program (OR: 1.96; 95%CI: 1.010−3.806; p = 0.02), HPV infection (OR: 7.35; 95%CI: 3.099−17.449; p< 0.001) or HPV test (OR: 1.67; 95%CI: 0.950−2.948; p = 0.03). Moreover, women who did not previously undergo a cervical cancer screening program were more likely to accept the new screening method (OR: 1.62; 95%CI: 0.878−3.015; p = 0.04). (4) Conclusions: Our results showed high acceptability rates of vaginal HPV self-sampling among participants.


Double-edged role of resource competition in gene expression noise and control.
Despite extensive investigation demonstrating that resource competition can significantly alter the deterministic behaviors of synthetic gene circuits, it remains unclear how resource competition contributes to the gene expression noise and how this noise can be controlled. Utilizing a two-gene circuit as a prototypical system, we uncover a surprising double-edged role of resource competition in gene expression noise: competition decreases noise through introducing a resource constraint but generates its own type of noise which we name as "resource competitive noise." Utilization of orthogonal resources enables retainment of the noise reduction conferred by resource constraint while removing the added resource competitive noise. The noise reduction effects are studied using three negative feedback types: negatively competitive regulation (NCR), local, and global controllers, each having four placement architectures in the protein biosynthesis pathway (mRNA or protein inhibition on transcription or translation). Our results show that both local and NCR controllers with mRNA-mediated inhibition are efficacious at reducing noise, with NCR controllers demonstrating a superior noise-reduction capability. We also find that combining feedback controllers with orthogonal resources can improve the local controllers. This work provides deep insights into the origin of stochasticity in gene circuits with resource competition and guidance for developing effective noise control strategies.


Circulating levels of pregnancy-associated plasma protein-A (PAPP-A) and human chorionic gonadotrophin (hCG) in intrauterine and extrauterine pregnancies.
Pregnancy-associated plasma protein-A (PAPP-A) and human chorionic gonadotrophin (hCG) have been measured in the plasma of pregnant and non-pregnant women attending the outpatient clinic for suspicion of pregnancy. The plasma concentrations obtained were grouped into intrauterine or extrauterine pregnancies and compared with values obtained in non-pregnant patients with similar periods of amenorrhoea. Patients with ectopic pregnancies had slightly lower PAPP-A levels and significantly lower hCG concentrations than those in women with normal intrauterine pregnancies. Non-pregnant women had very low hCG and PAPP-A levels compared with those in pregnant patients. These data suggest that in patients with extrauterine pregnancies the poorly sustained ectopic trophoblast is unable to produce normal concentrations of hCG and probably PAPP-A and that the slightly diminished levels of PAPP-A in ectopic pregnancies might be derived from a decidual production.


Lactobacillus lactis and Pediococcus pentosaceus-driven reprogramming of gut microbiome and metabolome ameliorates the progression of non-alcoholic fatty liver disease.
Although microbioa-based therapies have shown putative effects on the treatment of non-alcoholic fatty liver disease (NAFLD), it is not clear how microbiota-derived metabolites contribute to the prevention of NAFLD. We explored the metabolomic signature of Lactobacillus lactis and Pediococcus pentosaceus in NAFLD mice and its association in NAFLD patients. We used Western diet-induced NAFLD mice, and L. lactis and P. pentosaceus were administered to animals in the drinking water at a concentration of 109 CFU/g for 8 weeks. NAFLD severity was determined based on liver/body weight, pathology and biochemistry markers. Caecal samples were collected for the metagenomics by 16S rRNA sequencing. Metabolite profiles were obtained from caecum, liver and serum. Human stool samples (healthy control [n = 22] and NAFLD patients [n = 23]) were collected to investigate clinical reproducibility for microbiota-derived metabolites signature and metabolomics biomarker. L. lactis and P. pentosaceus supplementation effectively normalized weight ratio, NAFLD activity score, biochemical markers, cytokines and gut-tight junction. While faecal microbiota varied according to the different treatments, key metabolic features including short chain fatty acids (SCFAs), bile acids (BAs) and tryptophan metabolites were analogously restored by both probiotic supplementations. The protective effects of indole compounds were validated with in vitro and in vivo models, including anti-inflammatory effects. The metabolomic signatures were replicated in NAFLD patients, accompanied by the comparable levels of Firmicutes/Bacteroidetes ratio, which was significantly higher (4.3) compared with control (0.6). Besides, the consequent biomarker panel with six stool metabolites (indole, BAs, and SCFAs) showed 0.922 (area under the curve) in the diagnosis of NAFLD. NAFLD progression was robustly associated with metabolic dys-regulations in the SCFAs, bile acid and indole compounds, and NAFLD can be accurately diagnosed using the metabolites. L. lactis and P. pentosaceus ameliorate NAFLD progression by modulating gut metagenomic and metabolic environment, particularly tryptophan pathway, of the gut-liver axis.


Structure of Saccharomyces cerevisiae Rtr1 reveals an active site for an atypical phosphatase.
Changes in the phosphorylation status of the carboxyl-terminal domain (CTD) of RNA polymerase II (RNAPII) correlate with the process of eukaryotic transcription. The yeast protein regulator of transcription 1 (Rtr1) and the human homolog RNAPII-associated protein 2 (RPAP2) may function as CTD phosphatases; however, crystal structures of Kluyveromyces lactis Rtr1 lack a consensus active site. We identified a phosphoryl transfer domain in Saccharomyces cerevisiae Rtr1 by obtaining and characterizing a 2.6 Å resolution crystal structure. We identified a putative substrate-binding pocket in a deep groove between the zinc finger domain and a pair of helices that contained a trapped sulfate ion. Because sulfate mimics the chemistry of a phosphate group, this structural data suggested that this groove represents the phosphoryl transfer active site. Mutagenesis of the residues lining this groove disrupted catalytic activity of the enzyme assayed in vitro with a fluorescent chemical substrate, and expression of the mutated Rtr1 failed to rescue growth of yeast lacking Rtr1. Characterization of the phosphatase activity of RPAP2 and a mutant of the conserved putative catalytic site in the same chemical assay indicated a conserved reaction mechanism. Our data indicated that the structure of the phosphoryl transfer domain and reaction mechanism for the phosphoryl transfer activity of Rtr1 is distinct from those of other phosphatase families.


Formal Patient Complaints and Malpractice Events Involving Orthopedic Spine Surgeons: A Ten-Year Analysis.
Case-control study. To analyze patient complaints, potential risk, and malpractice events involving orthopedic spine surgeons over a 10-year period. Unsolicited patient complaints may be associated with risk management and malpractice events. We analyzed patient complaint, potential risk event, and malpractice event data for six orthopedic spine surgeons over a 10-year period. Patient complaints were analyzed and classified according to the Patient Complaint Analysis System. Baseline demographics were recorded for patients with complaints as well as the surgeons. A control group consisting of all patients seen by the six surgeons during the study period was created to identify patient and physician risk factors for formal patient complaints. Event rates (for complaints, risk, and malpractice events) were calculated by dividing the number of events by the total number of unique patients seen. There were 214 complaint designations among 202 patients with formal complaints, resulting in a complaint rate of 0.79%. Patients were most likely to complain about access and availability (35%) followed by care and treatment (32%). Of the 68 complaints regarding care and treatment, 34 were related to dissatisfaction with surgical outcome. Complications were identified in 26/34 cases. The malpractice event rate ranged from 0.06% to 0.65%. Patients who had surgery ( P < 0.0001) or a mental, behavioral, or neurodevelopmental disorder ( P = 0.0004) were more likely to file complaints compared with the control group. While infrequent, patient complaints against orthopedic spine surgeons are most related to access and availability. The rate of malpractice events varies widely between surgeons.


Entrainment in solution of an oscillating NADH oxidase activity from the bovine milk fat globule membrane with a temperature-compensated period length suggestive of an ultradian time-keeping (clock) function.
Entrainment in solution of an oscillating activity with a temperature compensated period of 24 min is described for a NADH oxidase (NOX) activity of the bovine milk fat globule membrane, a derivative of the mammary epithelial cell plasma membrane. The period of 24 min remained unchanged at 17 degrees C, 27 degrees C and 37 degrees C whereas the amplitude approximately doubled with each 10 degree C rise in temperature (Q(10)congruent with 2). The periodicity was observed with both intact milk fat globule membranes and with detergent-solubilized membranes, demonstrating that the oscillations did not require an association with membranes. The periodicity was not the result of instrument variation or of chemical interactions among reactants in solution. Preparations with different periodicities entrained (autosynchronized) when mixed. Upon mixing, the preparations exhibited two oscillatory patterns but eventually a single pattern representing the mean of the farthest separated maxima of the two preparations analyzed separately emerged. The cell surface NOX protein is the first reported example of an entrainable biochemical entity with a temperature-compensated periodicity potentially capable of functioning as an ultradian or circadian clock driver.


Specific electrolyte effects on hemoglobin in denaturing medium investigated through electro spray ionization mass spectrometry.
We examine Hofmeister specific ion effects of electrolytes added to protein solution under conditions minimizing electrostatic attraction between cations and positively charged protein. Hemoglobin (Hb) in aqueous solution at the denaturing pH = 2.7 is investigated in the presence of several metal chlorides, along with sodium and potassium bromides, iodides and thiocyanates, using electrospray ionization mass spectrometry (ESI-MS). Salt concentration was varied to maximize peak intensity and bell-shaped profile in the ESI-MS spectrum. The α-chain of myoglobin is identified as the main pattern of the ESI-MS spectra in all Hb-salt systems. Both peak intensity and quality of the bell-shaped profile of the protein spectrum decrease in the cation order: K+ > > Mg2+ > Li+ > > Na+ > Ca2+ ≈ Cs+ > Rb+ for Hb-Metal Chloride systems, and decrease in the anion order: Cl- > Br- > I- > SCN- for systems of both Hb-NaX and Hb-KX salts. To quantify salt addition effects two Hofmeister specific electrolyte parameters HS, and PS are proposed. HS is the mean (Hb-salt)/Hb peak intensity ratio, measured for the nine peaks used for ESI-MS spectra deconvolution, taken at the same m/z values of the Hb profile. PS is the ratio between HS standard deviation and HS, and provides a specific perturbation parameter measuring the loss of protein structure. These two Hofmeister parameters give clear evidence of the effects induced either by KCl, MgCl2 and LiCl that enhance protein peak intensity, or by NaBr, NaI, NaSCN and KSCN that induce the protein fragmentation, due to electrolyte-mediated dissociation.


The triad "dogs, conservation and zoonotic diseases" - An old and still neglected problem in Brazil.
The presence of domestic/free-ranging dogs in Brazilian protected areas and native vegetation fragments is an important problem, mainly because these animals pose a threat to wild species that live in such areas. In addition, dogs constantly circulate between wildlife environments and urban regions, acting as "bridges" in spillover events. Dogs are traditionally recognized as vectors of zoonoses, which are correct, but their roles as facilitating agents for the "jump" of pathogens from wild animals to humans (and vice versa) are sparsely debated. In this context, this work briefly describes the different roles of dogs in the dynamics and ecology of infectious diseases, using the Brazilian scenario as a study model.


Sequential bilateral bundle branch block during dofetilide, a new class III antiarrhythmic agent, in a patient with atrial fibrillation.
The mechanism of wide QRS complex tachycardias during dofetilide infusion was studied in a patient with atrial fibrillation. Endocardial recordings from the intraventricular conduction system showed that dofetilide caused "classic" aberrant conduction (Ashman phenomenon, typical QRS morphology) at high prematurity ratios (preceding interval = 1.78 x coupling interval--290), thus mimicking ventricular ectopy. In addition, there was frequent sequential bilateral bundle branch block, caused by a significant difference in preceding bundle-to-bundle intervals (mean difference +/- 1 SD: 74 +/- 26 msec). The present findings may prove helpful in the clinical assessment of wide QRS complex rhythms after dofetilide and possibly other "pure" Class III antiarrhythmics.


Deterioration in asthma control when subjects receiving fluticasone propionate/salmeterol 100/50 mcg Diskus are "stepped-down".
In this study, 647 subjects stable on fluticasone propionate/salmeterol Diskus 100/50 mcg BID (FSC) were randomized to continue FSC 100/50 mcg BID or "step down" to either fluticasone propionate (FP) 100 mcg BID, salmeterol (SAL) 50 mcg BID, or montelukast (MON) 10 mg once daily for 16 weeks. Overall asthma control significantly improved in the FSC group; whereas, "stepping down" to FP, SAL, or MON resulted in deterioration in asthma control, as determined by decreased measures of lung function and clinical features. This study provides support that treatment of both inflammation and smooth muscle dysfunction may be necessary to achieve and maintain asthma control in patients uncontrolled on ICS.


Detection of Coxiella burnetii by the immunoperoxidase technique.
An immunoperoxidase method using Coxiella (C.) burnetii-specific hyperimmune serum raised in rabbits, and swine anti-rabbit horseradish peroxidase conjugate was applied to visualize C. burnetii in BGM cell cultures. The technique proved to be highly specific and did not result in any unspecific background staining. The technique was applied successfully to study the organism during multiplication in vacuoles of epithelial cells and in the phagolysosomes of macrophages. Preliminary results further indicate that the technique may be useful for the detection of the pathogen in tissues and milk samples.


Effects of calcium on a cyclic nucleotide system in soleus motor nerve terminals.
The effects of increased extracellular calcium levels on the responses of the motor nerve terminal to agents that increased intracellular levels of cyclic AMP was studied on the in vivo cat soleus muscle preparation. The neural and muscle responses to intra-arterially administered NaF, an activator of adenylate cyclase, and dbcAMP progressively increased as blood calcium levels rose. These results provide additional support for the hypothesis of an interaction between calcium and cyclic AMP in motor nerve endings.


Usefulness of a cobalt chromium coronary stent alloy.
The cobalt chromium Guidant Multi-Link Vision coronary stent was deployed in 267 patients as part of a multicenter international registry. Major adverse cardiovascular events were infrequent, and late (180 days) quantitative angiography demonstrated binary (>50%) in-stent restenosis in 15.7% of patients. This registry establishes the safety and efficacy of this alloy as a coronary stent platform.


Mechanism of osteoclastic bone resorption: a new hypothesis.
Osteoclastic bone resorption involves the solubilization of the mineral salts and the degradation of noncollagen bone matrix and collagen fibrils. As no recognizable collagen fibrils have ever been reported within cytoplasmic vacuoles in osteoclasts, it is generally assumed that the collagen fibrils are digested extracellularly in the resorption zone. The extent to which lysis occurs extracellularly and whether or not the osteoclasts phagocytose the degradation products remain to be established. In the present communication, a hypothesis is presented suggesting the possibility that osteoclastic resorption of bone involves the participation of two different cell types. According to this hypothesis, osteoclastic bone resorption is initiated by osteoclasts that demineralize areas of bone and degrade noncollagen bone matrix. After the osteoclasts have moved away or become partially detached from the demineralized site, the exposed collagen fibrils are phagocytosed by mononuclear, fibroblast-like or monocyte-derived cells.


Cytology in the differential diagnosis of vaginitis.
Vaginal discharge, vulvar pruritus, and dyspareunia are among the most common symptoms that prompt women to seek medical advice. They are frequently assumed to be due to vaginitis resulting from infection and are treated with anti-infective agents without further investigation. Such empirical treatment may be successful, but on the frequent occasions when such treatment is inappropriate, the stage is set for deteriorating genital health. I believe that most cases of "vulvodynia" could be prevented by a correct diagnosis and treatment at the onset of symptoms.


Monitoring Activity-Dependent Bulk Endocytosis in Primary Neuronal Culture Using Large Fluorescent Dextrans.
The efficient recycling of synaptic vesicles (SVs) during neuronal activity is central for sustaining brain function. During intense neuronal activity, the dominant mechanism of SV retrieval is activity-dependent bulk endocytosis (ADBE). Here, we describe a method to monitor ADBE in isolation from other SV endocytosis modes, via the uptake of large fluorescent fluid-phase markers in primary neuronal culture. Furthermore, we outline how to monitor ADBE using this approach across a field of neurons or in individual neurons.


Suppressive effect of mirtazapine on the HPA system in acutely depressed women seems to be transient and not related to antidepressant action.
Impaired regulation of the hypothalamus-pituitary-adrenocortical (HPA) system is a consistent finding among patients with depression, which can be most sensitively detected with the combined dexamethasone (dex)/corticotrophin releasing hormone (CRH) test. The majority of patients with acute depression shows an exaggerated plasma corticotrophin (ACTH) and cortisol response to this test that normalizes gradually during successful antidepressant therapy. In contrast, persistently high HPA-responses to this challenge are prognostically less favorable. It has been recently questioned, whether this observation applies also to treatment with the atypical antidepressant mirtazapine, as patients treated with this drug showed a distinct attenuation of the endocrine response to the dex/CRH test already after 1 week of treatment. In the present study, we investigated whether the attenuating effect of mirtazapine on the HPA system is an acute pharmacological reaction disappearing after physiological adaptation or whether this effect is related to the antidepressant action of the drug. We examined plasma ACTH and cortisol responses to the dex/CRH test in acutely depressed inpatients treated either with mirtazapine (n=55) or a monoamine reuptake inhibitor (n=105) according to doctor's choice and compared the test results with healthy controls (n=40). Patients treated with monoamine reuptake inhibitors received either selective serotonin reuptake inhibitors (SSRI), tricyclic antidepressants (TCA) or the combined serotonin and noradrenalin reuptake inhibitor venlafaxine. We found increased plasma ACTH and cortisol responses to the dex/CRH test in depressed patients compared with healthy controls, but also significantly (p=.017) attenuated plasma cortisol secretion in the mirtazapine group compared to the group of monoamine reuptake inhibitor treated patients. This effect was not significant in male patients. Furthermore this effect was independent of the psychopathological state, but depended on treatment duration. Patient treatment with mirtazapine for up to 7 days resulted in dex/CRH test outcome that was indistinguishable from controls. This effect, however waned as it was not observable in patients treated for a longer period. These results suggest that short-term administration of mirtazapine has immediate but only transient suppressive effects on the HPA system predominantly in women. Our results confirm that dex/CRH tests can be used as predictors of clinical course also under mirtazapine treatment.


Preferences and non-preferences for nectar constituents inOrnithoptera priamus poseidon (Lepidoptera, Papilionidae).
Preferences for nectar sugars and amino acids ofOrnithoptera priamus butterflies were tested experimentally. Both male and female butterflies clearly preferred a sucrose solution over a glucose solution of equal concentration (25%, weight to total weight) and equally a fructose solution over a glucose solution. A significant trend of males to prefer fructose over sucrose and of females to prefer sucrose over fructose was detected. However, neither males nor females discriminated between a mimic ofLantana camara nectar containing amino acids and a corresponding plain sugar solution. These results suggest that butterflies select against glucose in floral nectar but do not support the hypothesis that butterflies select for high levels of amino acids in nectar. The rather unspecific response ofOrnithoptera priamus butterflies to the tested nectar constituents may reflect a generalist feeding strategy of these long-lived and spectacular butterflies.


Oral bioavailability of the antioxidant astaxanthin in humans is enhanced by incorporation of lipid based formulations.
Astaxanthin is a carotenoid with antioxidant properties, synthesised by plants and algae, and distributed in marine seafood. Astaxanthin is also available as a food supplement, but, like other carotenoids, is a very lipophilic compound and has low oral bioavailability. However, bioavailability can be enhanced in the presence of fat. There is not much information in the literature about the pharmacokinetics of oral astaxanthin in humans. In this open parallel study, healthy male volunteers received a single dose of 40 mg astaxanthin, as lipid based formulations or as a commercially available food supplement, followed by blood sampling for further analysis of plasma concentrations. Pharmacokinetic parameters were calculated to evaluate the extent and rate of absorption from each formulation. The elimination half-life was 15.9+/-5.3 h (n=32), and showed a mono-phasic curve. Three lipid based formulations: long-chain triglyceride (palm oil) and polysorbate 80 (formulation A), glycerol mono- and dioleate and polysorbate 80 (formulation B), and glycerol mono- and dioleate, polysorbate 80 and sorbitan monooleate (formulation C), all showed enhanced bioavailability, ranging from 1.7 to 3.7 times that of the reference formulation. The highest bioavailability was observed with formulation B, containing a high content of the hydrophilic synthetic surfactant polysorbate 80.


Photon correlation spectroscopy study on the stability of small unilamellar DPPC vesicles.
The growth in size of dipalmitoylphosphatidylcholine small unilamellar vesicles (SUV) below Tm has been studied by photon correlation spectroscopy and differential scanning calorimetry. We see an initial fast rise of the hydrodynamic diameter of the vesicles followed by a slower increase. We assign the slow component of the size change to fusion of SUV. The order of the kinetics appears to be higher than first order. The estimated half lifetime of the fusion is approximately 67 h. The diameters for the fast and slow processes at t = O are 756 and 256 A, respectively, while as t leads to infinity the diameters increase to 1,570 and 733 A, respectively.


Peripubertal diazepam administration prevents the emergence of dopamine system hyperresponsivity in the MAM developmental disruption model of schizophrenia.
Schizophrenia is believed to arise from an interaction of genetic predisposition and adverse environmental factors, with stress being a primary variable. We propose that alleviating anxiety produced in response to stress during a sensitive developmental period may circumvent the dopamine (DA) system alterations that may correspond to psychosis in adults. This was tested in a developmental rat model of schizophrenia based on prenatal administration of the mitotoxin methyl azoxymethanol acetate (MAM). MAM administration leads to a hyperdopaminergic state consisting of an increase in the number of DA neurons firing spontaneously, which correlates with an increased behavioral response to amphetamine. MAM-treated rats exhibited a heightened level of anxiety during adolescence. Peripubertal administration of the antianxiety agent diazepam was found to prevent the increase in DA neuron activity and blunt the behavioral hyperresponsivity to amphetamine in these rats. These data suggest that the pathophysiological factors leading to the onset of psychosis in early adulthood may be circumvented by controlling the response to stress during the peripubertal period.


Sodium hyaluronate's effect on xerophthalmia: a meta-analysis of randomized controlled trials.
Several studies in the past have attempted to demonstrate the efficacy of sodium hyaluronate in the treatment of xerophthalmia. However, results have been conflicting and a definite conclusion has not yet been reached. In order to provide integrated evidence for the effectiveness of sodium hyaluronate and to judge the methodological value of relevant randomized controlled trials (RCTs) in nearly thirty years, we conducted this meta-analysis. A range of electronic databases were searched: MEDLINE, the Cochrane Library Database, EMBASE, CINAHL, Web of Science and the Chinese Biomedical Database (CBM) without language restrictions. Two independent reviewers assessed trials for eligibility and quality, and meta-analysis was performed using the STATA 12.0 software. An integrated odds ratio (OR) with its corresponding 95% confidence interval (95% CI) was calculated. Six RCTs were included with a total of 839 xerophthalmia patients. The meta-analysis results revealed that patients with xerophthalmia who received the intervention of sodium hyaluronate eye drops didn't have significantly higher remission rate of dry eye symptoms than those in controlled groups (OR = 1.811, 95% CI = 0.741-4.429, p = 0.193). Sensitivity analysis suggested that the statistical results were robust. No publication bias was detected in this meta-analysis (p > 0.05). Although sodium hyaluronate can be used to help relieve the symptoms of dry eyes, present evidence cannot show in unequivocal terms that patients with xerophthalmia can benefit more from the clinical application of sodium hyaluronate than other eye drops or therapies.


Tissue Doppler Imaging predicts central sleep apnea in patients with chronic heart failure: data from the Daunia Registry.
Tissue Doppler imaging (TDI) is used to improve risk stratification in patients with chronic heart failure (CHF). So far, few studies have used this method to investigate the characteristics of subjects with CHF and Cheyne-Stokes breathing (CSB). The aim of this study was therefore to evaluate whether TDI assessment may predict the presence of CSB in patients with CHF. A total of 41 consecutive patients with CHF enrolled in the Daunia Heart Failure Registry underwent echocardiography assessment and nocturnal polygraphy to evaluate the presence of sleep apnea and CSB. Conventional echocardiography and TDI parameters were calculated. We have also quantified by TDI a combined index (EAS index) of diastolic and systolic performance: E'/(A' × S'). Subjects with evidence of CSB (N = 8) were characterized by lower values of A' (5·03 ± 2·64 vs. 7·88 ± 2·64 cm/s, P < 0·01). A' and EAS index values were related to Cheyne-Stokes episode rates (r = -0·49 and 0·52, P < 0·05 and <0·01 respectively), EAS index values also with the number of episodes of central apnea (r = 0·39, P < 0·05). A' values predicted the presence of CSB at poly-somnography examination with an OR 0·62 (95% CI 0·40-0·96, P < 0·05) even after correction for age and gender. Tissue Doppler imaging values (A') are associated with the presence of sleep apnea at nocturnal polygraphy.


The replication strategy of foamy viruses.
The replication strategy of foamy viruses diverges in many aspects from what is commonly accepted as the rules of retroviral replication. Although many questions on the details of the replication pathway are still unanswered, it appears that foamy viruses have adopted a strategy which functionally bridges the retroviral and the hepadnaviral replication pathways. A number of experimental findings in favour of the view that foamy viruses are reverse transcribing DNA viruses which integrate into the host cell genome are discussed.


Probability and informed consent.
In this paper, we illustrate some serious difficulties involved in conveying information about uncertain risks and securing informed consent for risky interventions in a clinical setting. We argue that in order to secure informed consent for a medical intervention, physicians often need to do more than report a bare, numerical probability value. When probabilities are given, securing informed consent generally requires communicating how probability expressions are to be interpreted and communicating something about the quality and quantity of the evidence for the probabilities reported. Patients may also require guidance on how probability claims may or may not be relevant to their decisions, and physicians should be ready to help patients understand these issues.


Ocular pain after intravitreal injection.
To evaluate the efficacy of different anesthetics and topical anti-inflammatory treatment in patients undergoing intravitreal injections (IVI). Prospective, randomized, double masked, comparative study. Patients undergoing 0.05 mL IVI were randomized to two different preoperative anesthetic regimes (regime A [0.5% tetracaine + naphazoline] versus regime B [5% lidocaine]) and two different post-injection topical protocols (protocol 1 [tobramycin qid] versus protocol 2 [tobramycin qid + diclofenac qid]). Patients were trained to score pain using a numerical rating pain scale from 0 (no pain) to 10 (excruciating pain) immediately after the injection, 30 min and 24 h later. Patients were instructed to take oral paracetamol (650-1000 mg, adjusted to the patient's weight) every six hours ad lib if necessary. A total of 156 patients were enrolled; 86 patients were randomized to regime A and 70 to regime B; 78 patients were assigned to each of the post-injection topical protocols. The average pain score immediately after the IVI was 2.77 (SD 2.12) for the whole group (2.85, SD 2.23 with tetracaine and 2.67, SD 2.00 with lidocaine; p = 0.73, Mann-Whitney U-test). Twenty-four hours later, the average pain score was 1.84, SD 2.45 (topical diclofenac + tobramycin) versus 1.75, SD 1.83 (topical tobramycin; p = 0.46, Mann-Whitney U-test). Forty-seven patients (30%) required oral paracetamol (average 3.3 and range 1-5 tablets). Conjunctival hemorrhage 30 min after the injection was less frequent and severe in eyes treated with topical naphazoline (p = 0.055, Mann-Whitney U-test). Topical tetracaine and lidocaine provide similar anesthesia before IVI. Topical diclofenac does not seem to reduce pain scores after IVI.


Structure-function relationships of endothelins, sarafotoxins, and their receptor subtypes.
The endothelins (ETs) and sarafotoxins (SRTXs) are two structurally related families of potent vasoactive peptides. Although their physiological functions have yet to be precisely elucidated, it seems likely that the ETs are involved in pathophysiological conditions such as hypertension and heart failure. In this minireview, recent advances in the biochemical characterization of the ET/SRTX system, with special reference to structure-function relationships and ET/SRTX receptor subtypes, are described, as well as the recent cloning and expression of ET receptors.


Personalized Antibodies for Gastroesophageal Adenocarcinoma (PANGEA): A Phase II Study Evaluating an Individualized Treatment Strategy for Metastatic Disease.
The one-year and median overall survival (mOS) rates of advanced gastroesophageal adenocarcinomas (GEA) are ∼50% and <12 months, respectively. Baseline spatial and temporal molecular heterogeneity of targetable alterations may be a cause of failure of targeted/immunooncologic therapies. This heterogeneity, coupled with infrequent incidence of some biomarkers, has resulted in stalled therapeutic progress. We hypothesized that a personalized treatment strategy, applied at first diagnosis then serially over up to three treatment lines using monoclonal antibodies combined with optimally sequenced chemotherapy, could contend with these hurdles. This was tested using a novel clinical expansion-platform type II design with a survival primary endpoint. Of 68 patients by intention-to-treat, the one-year survival rate was 66% and mOS was 15.7 months, meeting the primary efficacy endpoint (one-sided P = 0.0024). First-line response rate (74%), disease control rate (99%), and median progression-free survival (8.2 months) were superior to historical controls. The PANGEA strategy led to improved outcomes warranting a larger randomized study. SIGNIFICANCE: This study highlights excellent outcomes achieved by individually optimizing chemotherapy, biomarker profiling, and matching of targeted therapies at baseline and over time for GEA. Testing a predefined treatment strategy resulted in improved outcomes versus historical controls. Therapeutic resistance observed in correlative analyses suggests that dual targeted inhibition may be beneficial.This article is highlighted in the In This Issue feature, p. 211.


Polyethylene glycol embedding: a technique compatible with immunocytochemistry, enzyme histochemistry, histofluorescence and intracellular staining.
A technique is described which permits rapid processing of neural tissue for light microscopic analysis of sections of 1-40 microns thickness. This technique was developed as an alternative to paraffin embedding. When compared to paraffin, polyethylene glycol (PEG) offers the following advantages: 10-15 degrees C lower embedding temperatures, net tissue shrinkage of less than 5% vs approximately 50% in paraffin, and approximately one-half the embedding time. Tissue orientation during embedding and sectioning is particularly easy to control, e.g. 500 microns brain slices can be routinely flat-embedded and sectioned at 5 microns to form excellent ribbons. Since PEG is water-soluble, tissue may be dehydrated with a series of aqueous PEG solutions; the embedding matrix is easily removed by washing with a variety of aqueous buffers. These procedures allow subsequent electron microscopic analysis of material with generally well preserved ultrastructure. However, PEG is hygroscopic, thus tissue blocks become soft and difficult to section in high (greater than 90%) relative humidity. PEG was found to be compatible with intracellular staining with Lucifer yellow, horseradish peroxidase enzyme histochemistry, aqueous histofluorescence and immunocytochemical demonstration of neuronal peptides and glial fibrillary acidic protein.


Characteristic [3H]glucosamine-labeled glycoproteins in two-dimensional electrophoretograms of human renal cancer cells: identification as cathepsin D.
Two-dimensional electrophoretograms of extracts of [3H]glucosamine-labeled human renal cancer cells demonstrated a series of components (Mr 48,000 and 30,000) that are only poorly expressed in similarly labeled normal kidney epithelial cell cultures [S. Ogata, R. Ueda, and K. O. Lloyd (1981) Proc. Natl. Acad Sci. USA 78, 770-774]. These characteristics are also exhibited by [3H]Man-labeled samples and by concanavalin A-binding glycoproteins from [35S]Met-labeled cells. It is now shown that these species are the precursor chain (Mr 48,000) and native heavy chain (Mr 30,000) forms of the lysosomal enzyme, cathepsin D. These results were obtained by precipitation with a specific anti-cathepsin D serum and by binding of the components to pepstatin-Sepharose. Cathepsin D heavy chain is heterogeneous, having three major species with pI's of 5.7, 5.3, and 4.9; all forms are glycosylated with high mannose-type chains [approximate size: Man5(GlcNAc)2] and are partially phosphorylated. Despite these indications of dissimilarities in cathepsin D levels, the actual levels of total acid protease activity were not significantly higher in renal cancer cells than in normal kidney epithelial cells.


Validation of an Arabic version of the global Pelvic Floor Bother Questionnaire.
To validate an Arabic version of the global Pelvic Floor Bother Questionnaire (PFBQ), a self-administered 9-item symptom assessment tool. The translation-back translation method was used to create an Arabic version of the PFBQ. Clarity of terms and face validity were ensured by modifying the translated version according to comments from a focus group. The Arabic PFBQ was completed by 130 Lebanese women, who were allocated to affected (n=65) or control (n=65) groups on the basis of 2 verbally administered screening questions. Validity was assessed by comparing the PFBQ scores of the 2 groups. Test-retest reliability was evaluated in a subgroup of women who completed the PFBQ on 2 separate occasions; the intra-class correlation coefficient (ICC) for paired items was then calculated. Total PFBQ scores of the affected and control groups were significantly different (31.52 ± 16.80 vs 6.73 ± 7.50; P<0.001). Mean scores for individual PFBQ items were significantly different between the 2 groups, with the exception of dyspareunia. The ICC was above 0.7 for all individual items, thus confirming test-retest reliability. An Arabic version of the global PFBQ was developed and found to be both valid and reliable in the target population.


Testicular papillary cystadenomatous tumor of low malignant potential: a case report and discussion of the literature.
We report the fifth case of a rare testicular serous papillary cystadenomatous tumor of low malignant potential. All patients presented with a testicular mass. Histologically, the tumor can be differentiated from testicular papillary cystadenocarcinoma. The natural history of this rare tumor is yet to be determined.


Effects of cohort, sex, and birth date on body development of roe deer (Capreolus capreolus) fawns.
We investigated the effects of the year of birth (cohort effect), sex and birth date on the variations observed in birth weight and postnatal growth rate in 209 newborn fawns marked during eight consecutive fawning seasons from a highly productive roe deer population under good nutritional conditions. Roe deer exhibited a fast body development with a birth weight of 1628 g and a postnatal growth rate of 139 g/day constant over the first 20 days of their life. As previously reported for dimorphic and polygynous ungulates, there was a marked cohort effect on the rate of body development. This variability was partly explained by climatic conditions during late gestation. When fewer than 5 days with rainfall over 5 mm occurred in April, fawns were heavier. High temperatures during April and during the winter could also be involved in fast body development of roe deer fawns. Sex of fawn did not affect roe deer growth pattern. This was expected on the basis of low sexual dimorphism in size and low polygyny level characteristic of roe deer. Lastly, the date of birth did not affect the body development of roe deer fawns. High constancy of mean birth dates and high synchrony of births observed in this population could account for this result.


Perception is not all-purpose.
I aim to show that perception depends counterfactually on the action we want to perform. Perception is not all-purpose: what we want to do does influence what we see. After clarifying how this claim is different from the one at stake in the cognitive penetrability debate and what counterfactual dependence means in my claim, I will give a two-step argument: (a) one's perceptual attention depends counterfactually on one's intention to perform an action (everything else being equal) and (b) one's perceptual processing depends counterfactually on one's perceptual attention (everything else being equal). If we put these claims together, what we get is that one's perceptual processing depends counterfactually on one's intention to perform an action (everything else being equal).


Ultrasound salivary gland involvement in Sjogren's syndrome vs. other connective tissue diseases: is it autoantibody and gland dependent?
This study aims to investigate ultrasound (US) findings on salivary glands (SG) in patients with Sjögren syndrome (SS) vs. other connective tissue diseases (CTDs) and to assess the relationship of SGUS abnormalities with autoantibody profile in both groups. We enrolled 81 patients, 45 diagnosed with SS (39 with primary SS, 6 with secondary SS) and 36 diagnosed with other CTDs. All patients underwent a prospective evaluation of sicca symptoms, a Schirmer's test, and a B-mode US assessment of the parotid and submandibular glands, all blinded to the diagnosis. Each SG was semi-quantitatively scored 0-3; a grade ≥ 2 was considered pathological. SGUS involvement was classified as normal or pathological at the patient level and for each pair at the gland level. In addition, a total SGUS score of 0-12 and a parotid/submandibular score of 0-6 were calculated for each patient. Autoimmunity laboratory data were also obtained. All SGUS scores were higher in SS patients than in those with CTD (p < 0.001) and significantly more SS patients showed a pathological global (p < 0.001), parotid (p < 0.001), or submandibular (p = 0.001) US score compared with CTD patients. In SS patients, the presence of autoantibodies was significantly associated with pathological SGUS and higher scores, particularly at the parotid level, while in CTD patients, xerostomia and a pathological Schirmer's test were associated with pathological US and higher scores at the submandibular level (p < 0.05). SGUS showed a different grade of abnormality, site involvement, and associated autoantibody profile in SS patients as compared with other CTD. KEY POINTS: • Patients with SS and other CTDs showed different grades of SGUS abnormality. • Patients with SS and other CTDs showed different gland involvement and associated autoantibody profiles. • Anti-Ro60 and anti-Ro52 Ro60 positivity were associated with the severity of parotid involvement in SS patients.


Psychiatric hospitalization following psychosis onset: A retrospective cohort study using health administrative data.
There is limited evidence examining admissions in early psychosis. We sought to estimate the proportion of people with a psychiatric admission within 2 years of the first diagnosis of psychosis, and to identify associated risk factors. We constructed a cohort of incident non-affective psychosis cases using health administrative data and identified the first psychiatric hospitalization after psychosis onset. We compared hospitalization rates across sociodemographic, clinical and service-use factors. One in three patients had an admission within 2 years of first diagnosis. Younger age, migrant status, diagnosis of psychosis not otherwise specified, and prior substance use were associated with increased hospitalization rates, whereas family physician involvement in diagnosis was protective. Adolescents, immigrants and people presenting with diagnostic instability or prior substance use issues may benefit from interventions aimed at reducing hospitalization risk. Increasing primary care access and utilization among youth with early psychosis may also reduce hospitalization rates.


Aspects of feeding, including estimates of gut residence time, in three mytilid species (Bivalvia, Mollusca) at two contrasting sites in the Cape Peninsula, South Africa.
Individuals of three mytilid species (Choromytilus meridionalis; Perna perna; Aulacomya ater) from two sites characterised by different qualities of ration available to these suspension feeders showed different rates in some components of the physiological energy budget. These differences included higher feeding (=clearance) and respiration rates, but lower absorption efficiencies, in individuals from the site with the higher quality ration. A novel technique was employed to estimate the residence time of food particles in the digestive gland. Individuals showing higher feeding rates had shorter residence times than those feeding more slowly and a significant positive correlation was demonstrated between residence time and absorption efficiency. These relationships, together with an exponential increase in rates of respiratory heat loss with an increase in ingested ration, are suggested to provide these animals with a physiological flexibility to compensate for reduced food quality in a way consistent with some theoretical predictions. Such compensations are made more effective if the total gut capacity can also change in response to the quality of the ration.


[Geriatric rehabilitation care: Doing the right things right].
Geriatric rehabilitation concerns short-term integrated multidisciplinary care aimed at functional recovery and social participation for relatively frail elderly. Given the geriatric clients' complex care issues, nurses should possess sufficient and appropriate competencies in order to identify and assess the relevant symptoms and intervene effectively. Yet, nurses experience a certain apprehensiveness to perform their tasks and express difficulties in multidisciplinary communication and collaboration in a constructive manner. In addition to the client's and informal care giver's perception of their input in the geriatric rehabilitation process, this study provides an in-depth understanding of the way nurses perceive their role in geriatric rehabilitation. This descriptive study entails a quantitative and a qualitative component. The quantitative component concerns questionnaires for clients, informal care givers, nurses, and team leaders. The qualitative component aims to obtain in-depth information (i. e. opinions, meanings, and reflections) with regard to the decision making process and the performance of the rehabilitation care by means of open-ended questions (in the questionnaire) and semi-structured interviews. Clients and informal care givers rate specific themes in geriatric rehabilitation in a more negative light than nurses and team leaders do. These themes concern the provision of information in the hospital (prior to admission in the rehabilitation facility), involvement in the draw-up of the treatment plan and rehabilitation goals, geriatric rehabilitation as a 24/7 activity, and taking into account the client's other life events. The latter three findings in particular, are caused by nurses' apprehensiveness to perform their tasks adequately. Nurses working in geriatric rehabilitation, experience apprehensiveness to perform their tasks adequately. Uncertainty about the client's reaction or fear of damaging the relationship of trust, results in nurses not involving the clients and informal care givers in the draw-up of the rehabilitation goals. Apprehensiveness also submerges as the lack of experience or specific competences in considering the client's other life events. The recommendations address these aspects in particular.


Health-related quality of life and health utility for the institutional elderly in Taiwan.
To explore the health-related quality of life (HRQOL) and health utility in an institutional elderly population. Four hundred sixty-five elderly persons living in long-term care institutions in Taiwan were interviewed using Taiwan's abbreviated version of the World Health Organization Quality of Life (WHOQOL-BREF), rating scale (RS) and the Time-Trade-Off (TTO) utility measurement. The WHOQOL-BREF showed acceptable internal consistency (alpha range: 0.75-0.80 across domains) and validity. The sexual activity facet had the lowest response rate. Educational level, number of chronic diseases, physical performance, and number of caregivers had significant (p < 0.05) impacts on the domain scores of the WHOQOL-BREF. Physical performance had the strongest impact on the physical domain (R2 = 0.40) and accounted for significant percentages of the variance on the other three domains (R2 = 0.06-0.13). The mean RS score (score 0-100) was 61.3 +/- 16.2 (mean +/- SD). The mean TTO utility (score 0-1) was 0.92 +/- 0.22. Results indicate that the WHOQOL-BREF, excepting the sexual activity item, is useful for evaluating HRQOL of conscious elderly in institutions. The validity of TTO utility for studying the institutionalized elderly needs further evaluation.


Hemodynamic effects of anesthetic induction with eltanolone-fentanyl versus thiopental-fentanyl in coronary artery bypass patients.
We evaluated the hemodynamic profile of eltanolone and fentanyl versus thiopental and fentanyl anesthetic induction in patients with documented coronary artery disease. Fifty patients scheduled for coronary artery bypass grafting were randomly assigned to two treatment groups (25 patients each). Anesthesia was induced by eltanolone (0.5 mg/kg) or by thiopental (3 mg/kg). Each patient also received 3 micrograms/kg fentanyl and 0.1 mg/kg vecuronium. Heart rate, arterial, pulmonary arterial, central venous, and pulmonary capillary wedge pressures, and cardiac output were determined in the awake state, 2 min after induction of anesthesia, and at 1 and 5 min after intubation, which was performed 3 min after induction. Between-group statistics showed significantly (P < 0.05) lower mean arterial pressure and systemic vascular resistance for eltanolone-treated patients at all measuring points. Pulmonary capillary wedge pressure was lower at 1 min after intubation; left ventricular stroke work index was lower at 1 and 5 min after intubation in the eltanolone group. We conclude that the lower mean arterial pressure with eltanolone as compared to thiopental is a result of greater peripheral vasodilation.


Electrochemical characteristics of the immobilization of calf thymus DNA molecules on multi-walled carbon nanotubes.
Immobilization of DNA on carbon nanotubes plays an important role in the development of new types of miniature DNA biosensors. Electrochemical characteristics of the immobilization of calf thymus DNA molecules on the surfaces of multi-walled carbon nanotubes (MWNTs) have been investigated by cyclic voltammetry and electrochemical impedance analysis. The peak currents for Fe(CN)(6)(3-)/Fe(CN)(6)(4-) redox couple observed in the cyclic voltammograms decrease and the electron-transfer resistance (R(et)) obtained from the Nyquist plots increase due to the immobilization of DNA molecules (dsDNA or ssDNA) on the surfaces of MWNTs. Most of calf thymus DNA are covalently immobilized on MWNTs via diimide-activated amidation between the carboxylic acid groups on the carbon nanotubes and the amino groups on DNA bases, though the direct adsorption of the DNA molecules on MWNTs can be observed. Additionally, the interaction between DNA molecules immobilized on MWNTs and small biomolecules (ethidium bromide) can be observed obviously by cyclic voltammetry and electrochemical impedance analysis. This implies that the DNA molecules immobilized at the surface of MWNTs, with little structure change, still has the ability to interact with small biomolecules.


Multiple epiphyseal dysplasia: clinical and radiographic features, differential diagnosis and molecular basis.
Multiple epiphyseal dysplasia is one of the more common skeletal dysplasias but it can still be difficult to diagnose. The presenting signs are often rheumatological ('joint pain') or neurological ('myopathy') in nature, and the cardinal feature of skeletal dysplasia (short stature) may not be present. A radiographic skeletal survey is necessary to delineate the pattern of generalized delayed epiphyseal ossification and changes in epiphyseal contour. Once the diagnosis of multiple epiphyseal dysplasia has been established, careful examination of the radiographs can help to determine which genes should be analysed. Mutations in at least six different genes can cause multiple epiphyseal dysplasia, and it can be either dominant or recessive. Molecular diagnosis is important for accurate prognosis and genetic counselling.


The effects of hyperbaric oxygen on normal and ischemic colon anastomoses.
Leakage from colonic anastomoses is a major complication causing increased mortality and morbidity, and ischemia is a well-known cause of this event. Inadequate tissue oxygenation could be reversed by using hyperbaric oxygen. This study was designed to investigate the effects of hyperbaric oxygen on the healing of ischemic and normal colon anastomoses in the rat model. Standardized left colon resection 3 cm above the peritoneal reflection and colonic anastomosis were performed in 40 Wistar rats divided into four groups. The control group (I) received no further treatment. To mimic ischemia, 2 cm mesocolon was ligated on either site of the anastomosis in group II and IV rats. Hyperbaric oxygen therapy was started immediately after surgery in group III and IV rats (therapeutic groups). All animals were sacrificed on the fourth postoperative day. After careful relaparotomy, in situ bursting pressure was measured. The hydroxyproline contents of the anastomotic segments in equal length were determined. The hydroxyproline assay revealed that rats in group II with ischemic colonic anastomosis have significantly lower levels (P <0.05). The highest levels are in the group III rats with normal colonic anastomosis treated by hyperbaric oxygen (P <0.05). There was no significant difference in hydroxyproline levels between group II and group IV animals (P >0.05). Group III animals had significantly higher bursting pressures than any other group (P <0.05). Group II rats had lowest bursting pressures (P <0.05). Group IV animals had significantly higher levels than group II (P <0.05). Mean bursting pressure values both in groups III and IV and hydroxyproline levels in group III were significantly increased by hyperbaric oxygen therapy (P <0.05). Ischemia impairs anastomotic healing. Hyperbaric oxygen increases anastomotic healing of both normal and ischemic colonic anastomosis and reverses ischemic damage. This study demonstrated that hyperbaric oxygen improves anastomotic healing.


Correlation between RNA synthesis and ppGpp content in Escherichia coli during temperature shifts.
Both a correlation and a lack of correlation between guanosine 5'-diphosphate, 3'-diphosphate (ppGpp) level and RNA accumulation have been reported during temperature shifts of E. coli. We have reexamined these phenomena by measuring the total rate of RNA synthesis. After a temperature upshift (23 degrees to 40 degrees C) of E. coli relA+ and relA1 strains, there is an immediate increase in the rate of RNA synthesis which corresponds with the observed in vitro effects of temperature on RNA synthesis (Mangel 1974; Travers 1974). A subsequent increase in ppGpp level is correlated with a decrease in the rate of RNA synthesis. Conversely, following a temperature downshift (40 degrees to 23 degrees C), both relA+ and relA1 bacteria show an immediate decrease in the rate of RNA synthesis. Subsequently all strains studied decrease ppGpp content and correspondingly increase the rate of RNA synthesis after a downshift. By measuring the rate of RNA synthesis we have separated immediate temperature-induced changes in RNA synthesis, from the apparent effects of ppGpp during temperature shifts. As a result, during temperature upshifts and downshifts of relA+, and relA1 bacteria, an inverse correlation between ppGpp content and the total rate of RNA synthesis does exist. The fact that both relA+ and relA1 strains show similar responses to temperature shifts provides additional evidence for the function of relA-independent basal level ppGpp synthesis in regulating RNA synthesis in E. coli.


[Are there pseudophototropic reactions in biology? Part 4: On the reversibility of biologic/synthetic polymere systems (author's transl)].
In further research on pseudophototropic behaviour in cellular membranes of halobacteria the reversibility of vinylmethylethermaleic anhydride-copolymeres with biological liquids is tested and the basic principles of different colour generating reactions are studied.


Successful percutaneous aspiration thrombectomy for acute mesenteric ischemia in a patient with atrial fibrillation despite optimal anticoagulation therapy.
This report describes a case of acute mesenteric ischemia due to acute superior mesenteric artery (SMA) thromboembolism in a patient with chronic atrial fibrillation despite optimal anticoagulation therapy. The patient underwent SMA angiography and endovascular revascularization for acute mesenteric ischemia. We initially tried endovascular thrombolytic therapy for the thromboembolism of SMA, but it did not achieve complete revascularization. Therefore, we performed a percutaneous aspiration thrombectomy, which led to complete revascularization and successful treatment of acute mesenteric ischemia without any additional procedures.


Meeting report: transcranial magnetic stimulation and studies of human cognition.
A James S. McDonnell Foundation workshop examined the role of TMS in studies of human cognition. A summary of the workshop presentations, discussion, and the recommendations appear below. A selected reference list is provided at the end of the summary.


Colorimetric and electrochemical integrated dual-mode detection of glucose by utilizing CoOOH@Cu nanosheets as peroxidase mimetics.
Herein, we developed a colorimetric and electrochemical integrated dual-mode assay for glucose detection by utilizing CoOOH@Cu nanosheets as peroxidase mimetics. With the advantages of self-calibration, sensitivity and lower sample cost, this designed dual-mode assay offers great potential in blood glucose analysis.


Guanylate-binding proteins signature predicts favorable prognosis, immune-hot microenvironment, and immunotherapy response in hepatocellular carcinoma.
The role of guanylate-binding proteins (GBPs) in various cancers has been elucidated recently. However, our knowledge of the clinical relevance and biological characteristics of GBPs in hepatocellular carcinoma (HCC) remains limited. A total of 955 HCC patients were enrolled from five independent public HCC cohorts. The role of GBP molecules in HCC was preliminarily investigated, and a GBP family signature, termed GBPs-score, was constructed by principal component analysis to combine the GBP molecule values. We revealed the effects of GBP genes and GBPs-score in HCC via well-established bioinformatics methods and validated GBP1-5 experimentally in a tissue microarray (TMA) cohort. GBPs molecules were closely associated with the prognosis of patients with HCC, and a high GBPs-score highly inferred a favorable survival outcome. We also revealed high GBPs-score was related to anti-tumor immunity, the immune-hot tumor microenvironment (TME), and immunotherapy response. Among the GBPs members, GBP1-5 rather than GBP6/7 may be dominant in these fields. The TMA analysis based on immunohistochemistry showed positive correlations between GBP1-5 and the immune-hot TME with abundant infiltration of CD8+ T cells in HCC. Our integrative study revealed the genetic and immunologic characterizations of GBPs in HCC and highlighted their potential values as promising biomarkers for prognosis and immunotherapy.


Comparison of tomographic kidney volumes measured by semiautomatic segmentation method with scintigraphic split renal function in predicting posttransplant kidney functions.
In this study, it was aimed to compare scintigraphic split renal function (SRF) and computed tomographic (CT) kidney volumes by semiautomatic segmentation method in predicting graft functions after kidney transplantation. One hundred and twelve patients (77 males, 35 females) who had a living-donor kidney transplant between 2015 and 2017 in our centre were included in the study. While SRF was calculated with technetium-99m-diethylenetriaminepentaacetic acid (99m Tc-DTPA) scintigraphy, CT angiography was used for volumetric calculations. CT-volumetric measurements, especially renal cortical volume (RCV: 103.8 ± 20 ml) and ratio to body mass index (RCV/BMI: 4.45 ± 1.3) were found to be more significant than 99m Tc-DTPA-SRF in predicting graft functions. The correlations between SRF and RCV with 6th-month estimated glomerular filtration rate (eGFR) (rSRF: 0.052, rRCV: 0.317, p = 0.041) and 1st-year eGFR (rSRF: 0.104, rRCV: 0.374, p = 0.033) were found to be more significant in favour of RCV. The correlation between SRF/BMI and RCV/BMI with 1st-, 6th- and 12th-month eGFR (respectively, p = 0.02/0.048/0.024) were found to be more significant in favour of RCV/BMI. Although univariate analysis showed a significant relationship between most volumetric measurements and 1st-year graft functions, in multivariate analysis only RCV [odds ratio (OR): 1.04 (1.01-1.07), p = 0.023] and RCV/BMI [OR: 2.5 (1.27-5.39), p = 0.013] showed a significant relationship between graft functions. In our study, it was shown that CT-based renal volumetric measurements, especially RCV and RCV/BMI, predicted graft functions more strongly than scintigraphic 99m Tc-DTPA-SRF.


Discovery of spirooxadiazoline oxindoles with dual-stage antimalarial activity.
Malaria remains a prevalent infectious disease in developing countries. The first-line therapeutic options are based on combinations of fast-acting artemisinin derivatives and longer-acting synthetic drugs. However, the emergence of resistance to these first-line treatments represents a serious risk, and the discovery of new effective drugs is urgently required. For this reason, new antimalarial chemotypes with new mechanisms of action, and ideally with activity against multiple parasite stages, are needed. We report a new scaffold with dual-stage (blood and liver) antiplasmodial activity. Twenty-six spirooxadiazoline oxindoles were synthesized and screened against the erythrocytic stage of the human malaria parasite P. falciparum. The most active compounds were also tested against the liver-stage of the murine parasite P. berghei. Seven compounds emerged as dual-stage antimalarials, with IC50 values in the low micromolar range. Due to structural similarity with cipargamin, which is thought to inhibit blood-stage P. falciparum growth via inhibition of the Na + efflux pump PfATP4, we tested one of the most active compounds for anti-PfATP4 activity. Our results suggest that this target is not the primary target of spirooxadiazoline oxindoles and further studies are ongoing to identify the main mechanism of action of this scaffold.


Polymorphism of the X-chromosome, Y-chromosome and autosomes in the Australian hopping mice, Notomys alexis, N. cervinus and N. fuscus (rodentia, muridae).
All three species of Notomys so far studied possess a diploid number of 48. Many elements in the karyotype of N. alexis are polymorphic due to variation in heterochromatin, but the variation is most marked in autosomal pair 1, which occurs in at least four forms, the X-chromosome, which occurs in three forms, and the Y-chromosome which occurs in many forms. N. cervinus is unique in the genus in possessing an entirely biarmed karyotype due mainly to the addition of heterochromatic short arms. The X-chromosome of N. cervinus occurs in three forms and the Y-chromosome in two forms. The karyotype of N. fuscus is mainly telocentric although two autosomal pairs are polymorphic due to pericentric inversions. The X- and Y-chromosomes both occur in two forms in N. fuscus. Chromosome measurements and C-banding show that most of the variation in the size and morphology of the sex-pair both within and between species is due to variation in constitutive heterochromatin.


Multilocular renal cyst. Scanning and transmission electron microscopic observations.
A multilocular renal cyst in a boy aged one year and four months is presented, with particular attention being paid to the nature of the epithelial lining cells. Light and transmission electron microscopy showed the cyst to be lined by a single layer of flattened or cuboidal epithelial cells of relatively uniform morphology. Neither embryonic elements nor nephroblastomatous foci were noted in the intervening stroma. The scanning electron microscopy showed hitherto undescribed surface morphological features of the epithelial lining cells: They were characterized by the presence of one or, occasionally two centrally positioned long cilia and by variably oriented microvilli. The observations presented here suggested that the lining cells of the cyst most closely resembled the principal cells of the collecting ducts, especially those located in the inner medulla of the kidney. An unexpected finding was the additional occurrence of a giant bullous lesion in the right lung of this patient.


Hyperglycemia-induced downregulation of apolipoprotein M expression is not via the hexosamine pathway.
We previously demonstrated that hyperglycemia could suppress apolipoprotein M (apoM) synthesis both in vivo and in vitro; however, the mechanism of hyperglycemia-induced downregulation of apoM expression is unknown yet. In the present study we further examined if hexosamine pathway, one of the most important pathways of glucose turnover, being involved in modulating apoM expression in the hyperglycemia condition. We examined the effect of glucosamine, a prominent component of hexosamine pathway and intracellular mediator of insulin resistance, on apoM expression in HepG2 cells and in rat's models. In the present study we also determined apolipoprotein A1 (apoA1) as a control gene. Our results demonstrated that glucosamine could even up-regulate both apoM and apoA1 expressions in HepG2 cell cultures. The glucosamine induced upregulation of apoM expression could be blocked by addition of azaserine, an inhibitor of hexosamine pathway. Moreover, intravenous infusion of glucosamine could enhance hepatic apoM expression in rats, although serum apoM levels were not significantly influences. It is concluded that both exogenous and endogenous glucosamine were essential for the over-expression of apoM, which may suggest that the increased intracellular content of glucosamine does not be responsible for the depressed apoM expression at hyperglycemia condition.


Atmospheric nitrate formation pathways in urban and rural atmosphere of Northeast China: Implications for complicated anthropogenic effects.
Effects of human activities on atmospheric nitrate (NO3-) formation remain unclear, though the knowledge is critical for improving atmospheric chemistry models and nitrogen deposition reduction strategies. A potentially useful way to explore this is to compare NO3- oxidation processes in urban and rural atmospheres based upon the oxygen stable isotope composition of NO3- (Δ17O-NO3-). Here we compared the Δ17O-NO3- from three-years of daily-based bulk deposition in urban (Shenyang) and forested rural sites (Qingyuan) in northeast China and quantified the relative contributions of different formation pathways based on the SIAR model. Our results showed that the Δ17O in Qiangyuan (26.2 ± 3.3‰) is significantly higher (p < 0.001) than in Shenyang (24.0 ± 4.0‰), and significantly higher in winter (Shenyang: 26.1 ± 6.7‰, Qingyuan: 29.6 ± 2.5‰) than in summer (Shenyang: 22.7 ± 2.9‰, Qingyuan: 23.8 ± 2.4‰) in both sites. The lower values in the urban site are linked with conditions that favored a higher relative contribution of nitrogen dioxide reaction with OH pathway (0.76-0.91) than in rural site (0.47-0.62), which should be induced by different levels of human activities in the two sites. The seasonal variations of Δ17O-NO3- in both sites are explained by a higher relative contribution of ozone-mediated oxidation chemistry and unfavorable conditions for the OH pathway during winter relative to summer, which is affected by human activities and seasonal meteorological condition change. Based on Δ17O, wintertime conditions led to a contribution of O3 related pathways (NO3 + DMS/HC and N2O5 hydrolysis) of 0.63 in Qingyuan and 0.42 in Shenyang, while summertime conditions led to 0.15 in Qingyuan and 0.05 in Shenyang. Our comparative study on Δ17O-NO3- between urban and rural sites reveals different anthropogenic effects on nitrate formation processes on spatial and temporal scales, illustrating different responses of reactive nitrogen chemistry to changes in human activities.


Efficacy and bleeding risk of antithrombin supplementation in septic disseminated intravascular coagulation: a prospective multicenter survey.
Although supplementation with antithrombin (AT) concentrates has been widely accepted for the treatment of disseminated intravascular coagulation (DIC) in Japan, the effects and adverse effects have not been investigated. We conducted a nonrandomized multi-institutional survey. A total of 729 septic DIC patients with AT activity levels of 70% or lower, who had undergone AT substitution at either 1500 IU/day or 3000 IU/day for consecutive 3 days were analyzed. Of these, 650 and 79 patients had received 1500 IU/day (AT1500 group) and 3000 IU/day (AT3000 group), respectively. Bleeding events were observed in 6.52% of patients (severe bleeding, 1.71%). A significant decrease in initial AT level (below 50%) was observed in 69.6% of patients in AT3000 group and 48.2% in AT1500 group, and this difference was significant (P<0.01). A logistic-regression analysis conducted using age, gender, body weight, initial AT activity, and supplemented AT dose, revealed that higher initial AT activity (odds ratio (OR), 1.032; P<0.001), AT dose of 3000 IU/day (OR, 1.912; P=0.026), and age (OR, 0.985; P=0.023) were significant factors for improved survival. The risk of severe bleeding is less than 2%, and concomitant administration of heparin did not increase the risk. The survival in AT1500 group was 65.2%, while that in AT3000 group was 74.7%.


Assessing students' mental health in two American dental hygiene programs.
Dental hygiene students adapt to new environments while learning technical skills and providing clinical care during their education. Understanding how stress affects students in their professional healthcare program warrants exploration. This study assessed stress among dental hygiene students in two educational settings in the Southeastern United States. First and second-year dental hygiene students (N = 136) from a community college setting (n = 67) and a university setting (n = 69) were invited to complete an anonymous online survey on mental health in fall 2019. Validated surveys on depression, anxiety, social support, and burnout were included. Data analysis included chi-squared and Mantel-Haenszel statistics, depending on the scale of measurement, with the level of significance set at 0.05 for all analyses. Participants included 54 dental hygiene students from a community college (Response rate = 80.6%) and 69 dental hygiene students from a university (RR = 100%). There was a statistically significant difference in the proportion of students reporting moderately severe or severe anxiety (p = 0.007), with 56% (n = 30) of the community college respondents and 36% (n = 24) of the university reporting these anxiety levels. Students attending a community college were also more likely to express feelings of worry (n = 34) compared to students in a university setting (p = 0.005). There was no statistically significant difference in depression (p = 0.07) or suicidal thoughts (p = 0.41). Dental hygiene students enrolled in these two programs reported high levels of self-reported stress, mental and emotional concerns that may increase suicidal tendencies.


Transient first-degree AV block and sixth nerve palsy in a patient with closed head injury.
We present the case of an 8-year-old girl who suffered a closed head injury with the development of bilateral sixth-nerve palsies, and who was noted upon arrival to the emergency department to be in first-degree AV block. The sixth-nerve palsies and the first-degree AV block resolved simultaneously within 24 hours of injury. We discuss dysrhythmias and ECG changes previously reported with central nervous system diseases and their proposed etiology.


Infection as a contributory cause of death in patients hospitalized for motor vehicle trauma.
The age and length-of-stay specific incidence of fatal infections and the percentage of deaths due to infection were determined in a population-based sample of patients hospitalized for more than 2 days for motor vehicle injuries. The percentage of deaths due to infection was determined from a statewide sample of death certificates, and the patient fatality rates for hospitalized motor vehicle crash injury patients were determined from statewide Professional Activities Study data. Fatal pulmonary infections were far more common than fatal nonpulmonary infections. The incidence rate of fatal pulmonary infection was highest among those with hospital stays of 7 to 28 days and among those 70 years of age or older. Incidence rates varied substantially by age and length of hospital stay from a low of 8 deaths per 100,000 patient-days for adult female patients with stays longer than 28 days to a high of 102 deaths per 100,000 patient-days for elderly men with stays of 7 to 28 days. The incidence rates documented in this study may improve the identification of blunt trauma patient groups with the greatest need for prophylactic antiinfective measures.


Seroprevalence of antibodies against SARS-CoV-2 in the Valencian Community (Spain) as of October 2022.
To estimate the seroprevalence of SARS-CoV-2 antibodies in the Valencian Community (Spain) in October 2022, when BA.5 was the predominant variant. Cross-sectional, region-wide, population-based serosurvey study in 88 randomly selected primary care centers of the Valencian Community. Seroprevalence of anti-nucleocapsid (indicative of past infection) and total receptor binding domain (indicative of past infection or vaccination) antibodies was 71.0% (confidence interval [CI]: 67.8-74.2) and 98.4% (CI: 97.5-99.3), respectively. 66.7% (CI: 63.4-70.0) of the population shows hybrid immunity, but only 43.2% in those 80 and over. The high proportion of hybrid immunity detected is relevant for public health strategies. A second vaccination booster was advisable in the elderly population.


A Health Education Intervention as the Control Condition in the CTN-0037 STRIDE multi-site exercise trial: Rationale and Description.
The selection of a control condition in a randomized controlled trial (RCT) is critical in determining the effect of the experimental treatment. While the use of a placebo pill can be an ideal control in pharmaceutical trials, RCTs of behavioral interventions present unique challenges in the selection and implementation of the appropriate control condition. Investigators must not only consider the control condition's ability to protect against threats to internal validity and its plausibility as a possible intervention, but must also carefully implement the control condition so it does not introduce bias from either the investigators or the participants. The purpose of this paper is to provide the rationale for the use of a health education intervention (HEI) as the control condition in the CTN-0037 Stimulant Reduction Intervention Using Dosed Exercise (STRIDE) trial. In this paper, we will describe the careful design of the HEI to ensure proper implementation and discuss alternative control conditions considered.