Antibacterial properties of hexanal-chitosan nanoemulsion against Vibrio parahaemolyticus and its application in shelled shrimp preservation at 4 °C.
Vibrio parahaemolyticus has been considered as the leading pathogen associated with seafood-borne disease. Hexanal possesses antibacterial property but the hydrophobicity and volatility limit its application. The purpose of this study was to prepare hexanal-chitosan nanoemulsion (HCN), investigate its antibacterial ability against V. parahaemolyticus, and examine the combination of HCN with sodium alginate coating on the quality attributes of shrimp during cold storage. The mean droplet size of HCN fabricated by ultrasonic emulsification was 91.28 nm. HCN showed regular spherical shape and exhibited good centrifugation stability and storage stability at 4 °C. HCN exerted anti-V. parahaemolyticus effect with the minimum inhibitory concentration and minimal bactericidal concentration of both 5 mg/mL. Furthermore, HCN induced morphological changes and destroyed bacterial membrane, resulting in cell death. The results of preservation test showed that HCN alone and its combination with sodium alginate coating effectively retarded the quality deterioration and microbial spoilage of shelled shrimps during refrigerated storage. Comparatively, the combination treatment exhibited better preservation effect. The present study suggested that HCN prepared by ultrasonic emulsification is an effective alternative to control V. parahaemolyticus contamination in seafood and also shows great application potential in the quality maintaining of seafood during cold storage.


A systematic review of in-hospital worsening heart failure as an endpoint in clinical investigations of therapy for acute heart failure.
In-hospital worsening heart failure (WHF) occurs frequently in patients hospitalized for acute heart failure (AHF) and has strongly negative prognostic associations. It may be a useful endpoint in studies of AHF management but important questions remain regarding optimization of its definition and variability in its incidence. Our objective was to survey the full extent of clinical interest in WHF and assess the impact of baseline variables and trial design on outcomes. PubMed, Embase, and BIOSIS were searched systematically for clinical studies that had in-hospital WHF as an endpoint. Differences in definitions of in-hospital WHF were reviewed for their potential impact on observed incidence of WHF and its associations with post-discharge outcomes. The search identified 35 publications representing 13 interventional trials, 3 observational studies, several different classes of therapeutic agent, and 78,752 patients overall. Incidence of in-hospital WHF varied greatly-from 4.2% to 37%. Concerning the impact of differences in the way in which WHF was defined, two important factors were physician determination of worsening and whether intensification of diuretic therapy alone was defined as a WHF event. Patients having in-hospital WHF were at substantially greater risk for death and longer length of stay during index hospitalizations, all-cause and heart-failure rehospitalization, cardiovascular complications, renal failure, all-cause death, cardiovascular death, and higher healthcare costs post-discharge. There is diverse interest in selecting in-hospital WHF as an endpoint in clinical trials. Differences in reported incidence are complexly related to differences in the way in which WHF is defined.


Comparison of commercial diagnostic tests for identification of serogroup antigens of Neisseria meningitidis.
In the study that is described the sensitivities and specificities of three commercial tests and the standard Reference Laboratory test, used since 1961, to identify Neisseria meningitidis serogroups were compared. The tests marketed by Difco, Murex/Wellcome, and Sanofi/Pasteur showed overall sensitivities of 92, 95, and 100%, respectively, and specificities of 67, 88, and 82%, respectively. When limited to the common serogroups A, B, and C, the three tests yielded sensitivities of 93, 97, and 100%, respectively, and specificities of 98, 100, and 98%, respectively. However, determination of the uncommon serogroups X, W-135, Y, Z, and 29E with these tests is either unreliable or not possible.


Horizontal (vs. vertical) closure of the neo-pharynx is associated with superior postoperative swallowing after total laryngectomy.
We conducted a cross-sectional study to compare the horizontal and vertical methods used in the surgical closure of the neo-pharynx after total laryngectomy in terms of their effect on swallowing function, swallowing-related quality of life (QOL), and overall QOL. We also assessed the potential influence of age (≤64 vs. ≥65 yr) and the type of treatment modality (primary, salvage, or total laryngectomy with radiotherapy) on outcomes. Our final study population was made up of 34 patients-31 men and 3 women, aged 49 to 89 years (mean: 66.8)-who had undergone a total laryngectomy. One year after surgery, all patients were asked to complete the M.D. Anderson dysphagia inventory (MDADI), which quantifies swallowing function and swallowing-related QOL, and the University of Washington quality-of-life questionnaire (UW-QOL), which quantifies overall QOL. Of the 34 patients, 16 had undergone a horizontal surgical closure of their neo-pharynx and 18 a vertical closure. According to the MDADI, patients in the horizontal group experienced significantly better swallowing function/QOL; the mean composite MDADI scores were 91.5 in the horizontal group and 68.3 in the vertical group (p = 0.005). We found no significant difference in terms of overall QOL, as the respective mean UW-QOL scores were 81.0 and 80.8 (p = 0.93). The population correlation coefficient was positive in both groups, but more so in the horizontal group (ρhorizontal = 0.876 and ρvertical = 0.676). Neither age nor the type of treatment modality employed influenced swallowing function/QOL (page = 0.10, ptreatment modality = 0.78) or overall QOL (page = 0.08, ptreatment modality = 0.59). We conclude that horizontal closure of the neo-pharynx is superior to vertical closure in terms postoperative swallowing function/QOL but not overall QOL.


Pollution halo impact in context of productive capacities, energy poverty, urbanization, and institutional quality.
The Belt and Road Initiative (BRI) represents a substantial development strategy spearheaded by China. Its central aim is to foster connectivity across a vast geographical area that includes countries spanning Asia, Europe, and Africa. This project played a pivotal role to develop the region on the one side and also raised serious environmental concerns on the other side. There is extensive literature explored the various dimensions affecting the environment in BRI partner countries but there is hardly any study examining the impact of productive capacities, energy poverty, FDI, urbanization, and institutional quality on CO2 emission in the BRI region. Moreover, pollution halo impact is also explored so this study used panel data of 52 nations engaged in the BRI covering time span of 2001-2022 by applying OLS, Difference GMM, System GMM, Cross sectional-ARDL techniques. The results suggest that enhancing productive capacities, FDI and institutional quality significantly reduces carbon emissions in the region, while energy poverty, urbanization and economic growth is linked to higher carbon emissions. Moreover, 'pollution halo effect' is proved because of adoption of eco-friendly technologies through foreign corporations lead to reduction in carbon emission. The study advocates for policy measures that emphasize the promotion of productive capacities, the utilization of renewable energy sources, the adoption of practices regarding sustainable urban development, the implementation of efficient institutional structure, and inflow of eco-friendly technology through FDI.


A rotating disk sorptive extraction based on hydrophilic deep eutectic solvent formation.
An elegant preconcentration method assumed sorption of polar analytes from complex non-polar matrices on a rotating disk based on hydrophilic deep eutectic solvent formation is presented for the first time. The surface of poly(vinylidene fluoride-co-tetrafluoroethylene) rotating disk was coated with choline chloride acted as a precursor of deep eutectic solvent (hydrogen bond acceptor). The rotating disk was immersed in vegetable oil sample and phenolic compounds (hydrogen bond donors) were efficient separated on the disk during its rotation due to deep eutectic solvent formation. Ability of hydrophilic deep eutectic solvent decomposition in aqueous phase was used for fast analytes elution from the disk surface (2 min). Finally, the obtained aqueous solution of phenolic compounds and choline chloride was analyzed by high-performance liquid chromatography with fluorescence detection. Under optimal conditions, the limits of detection for gallic acid, protocatechuic acid, tyrosol, vanillic acid, p-coumarinic acid, syringaldehyde and thymol were in the range of 10-60 μg L-1. The developed approach allowed to significantly reduce sorption and elution time in comparison with previously reported rotating disk sorptive extraction approaches. The extraction mechanism based on deep eutectic solvent formation provided selective separation of target analytes with absolute extraction recovery in the range of 66-87%.


[Clinical reasoning and decision making in the practice. A woman with abdominal complaints].
A female patient aged 28 years suffered from recurrent episodes of diarrhoea. Giardia lamblia was isolated once. By rigorous application of basic clinical skills, such as thorough history taking and physical examination, together with laboratory tests the correct diagnosis was established (in this patient systemic lupus erythematosus). Two important aspects of clinical reasoning, viz. returning to the patient when new information becomes available (cyclic way of working) and pattern recognition ensured a fruitful diagnostic process.


Post-polymerization functionalization of poly(3,4-propylenedioxythiophene) (PProDOT) via thiol-ene "click" chemistry.
The surface functionalization of conjugated polymers such as the poly(alkoxythiophenes) poly(3,4-ethylenedioxythiophene) (PEDOT) and poly(3,4-propylenedioxythiophene) (PProDOT) provides a potential means for systematically tailoring their physical properties. We previously reported the synthesis of an alkene-functionalized 3,4-propylenedioxy-thiophene (ProDOT) derivative that could be readily modified through thiol-ene "click" chemistry. Here, we investigated the post-polymerization modification of PProDOT surfaces by using a dialkene functionalized variant (ProDOT-diene). The chemical structure of the ProDOT-diene monomer was confirmed by Nuclear Magnetic Resonance (NMR) and Fourier Transform Infrared spectroscopy (FTIR). The ProDOT-diene monomer was either chemically or electrochemically polymerized into the PProDOT-diene polymer, and then subsequently modified with alkyl, PEG, or ferrocene moieties via radical-based thiol-ene chemistry. We found that the normally insoluble PProDOT-diene could be converted into a soluble derivative by grafting alkyl groups onto the polymer chains after chemical polymerization. When electrochemically deposited on indium-tin oxide (ITO) glass electrodes, the conductivity, electroactivity and contact angles of the modified PProDOT-diene films could be tuned over a broad range. Scanning Electron Microscopy (SEM) revealed that post-polymerization modification did not significantly alter the surface morphology of the PProDOT-diene films. Overall, this method allows for efficient, facile tuning of the surface chemistry of poly(alkylthiophene) films, making it possible to tailor properties such as conductivity and wettability for different applications.


Role of RAGE in Alzheimer's Disease.
Receptor for advanced glycation end products (RAGE) is a receptor of the immunoglobulin super family that plays various important roles under physiological and pathological conditions. Compelling evidence suggests that RAGE acts as both an inflammatory intermediary and a critical inducer of oxidative stress, underlying RAGE-induced Alzheimer-like pathophysiological changes that drive the process of Alzheimer's disease (AD). A critical role of RAGE in AD includes beta-amyloid (Aβ) production and accumulation, the formation of neurofibrillary tangles, failure of synaptic transmission, and neuronal degeneration. The steady-state level of Aβ depends on the balance between production and clearance. RAGE plays an important role in the Aβ clearance. RAGE acts as an important transporter via regulating influx of circulating Aβ into brain, whereas the efflux of brain-derived Aβ into the circulation via BBB is implemented by LRP1. RAGE could be an important contributor to Aβ generation via enhancing the activity of β- and/or γ-secretases and activating inflammatory response and oxidative stress. However, sRAGE-Aβ interactions could inhibit Aβ neurotoxicity and promote Aβ clearance from brain. Meanwhile, RAGE could be a promoting factor for the synaptic dysfunction and neuronal circuit dysfunction which are both the material structure of cognition, and the physiological and pathological basis of cognition. In addition, RAGE could be a trigger for the pathogenesis of Aβ and tau hyper-phosphorylation which both participate in the process of cognitive impairment. Preclinical and clinical studies have supported that RAGE inhibitors could be useful in the treatment of AD. Thus, an effective measure to inhibit RAGE may be a novel drug target in AD.


A systematic review and meta-analysis of tolerability, cardiac safety and efficacy of inclisiran for the therapy of hypercholesterolemic patients.
Dyslipidaemia is a crucial risk factor for cardiovascular morbidity and mortality. A short interfering RNA called inclisiran diminishes circulating levels of PCSK9 and LDL-C by hindering PCSK9 translation in the liver. RCTs were electronically searched on PubMed, Cochrane Central, and Clinicaltrials.gov to assess the safety and efficacy of inclisiran. Cochrane Review Manager 5 was used to conduct the pooled analysis. Risk of bias was assessed and GRADE pro-GDT was utilized, respectively, to estimate the methodological quality and overall quality of evidence. Of 218 records screened, four studies were included with 2203 participants in inclisiran and 1949 participants in the placebo group. Inclisiran was related to non-significant elevated risk of total adverse events[RR = 1.05(0.98,1.12), p = 0.16; I2 = 53%], non-serious adverse events[RR = 1.09(0.97,1.22),p = 0.15;I2 = 61%] and all-cause mortality[RR = 1.01(0.60,1.70),p = 0.97;I2 = 0%] whereas a lower risk of serious adverse events[RR = 0.94(0.70,1.25),p = 0.67;I2 = 73%], cardiac disorders [RR = 0.87(0.66,1.15),p = 0.33;I2 = 42%] and Major adverse cardiovascular events(MACE)[RR = 0.79(0.62,1.00),p = 0.05; I2 = 0%] as compared to placebo. Inclisiran was also linked to a substantial decline in the percentage of LDL-C, PCSK9, total cholesterol, and Apo B. The pooled analysis of the existing evidence shows that inclisiran showed reduced risk of MACE along with excellent efficacy in managing dyslipidemia.


Update on sentinel lymph node biopsy after neoadjuvant chemotherapy in breast cancer patient.
Today, sentinel lymph node biopsy (SLNB) is considered the gold standard for axillary staging in early breast cancer patients with clinically negative lymph nodes (cN0). SLNB allows to determine the axillary lymph node status sparing the axillary dissection (AD) and its potential complications (seroma formation, loss of sensation, shoulder dysfunction and lymphedema) On the other hand, SLNB for nodal staging in breast cancer patients with clinically negative lymph nodes after neoadjuvant chemotherapy (ycN0) is a highly debated topic due to different reported success rates. In order to optimize oncological results, high identification rate (> 90%) and false negative rate as low as possible (< 10%) should always be obtained when performing SLNB after neoadjuvant chemotherapy. The success rates of SLNB after neoadjuvant chemotherapy (NAC) mainly depend on the clinical lymph node status pre-NAC. In patients with pre-NAC clinically negative nodes (cN0) and at restaging with post-NAC clinically negative nodes (ycN0), SLNB after chemotherapy should be performed because it is an accurate and safe procedure. In patients with pre-NAC clinically positive nodes (cN+) and at restaging with post-NAC clinically negative nodes (ycN0), SLNB after chemotherapy might be considered thanks to the high lymph nodal pathologic complete response rate; however, in this last setting, individual ability, technical skills and repetitive performance of specific tasks must always be followed to improve the identification rate and false negative rate. AD may be avoided only if sentinel lymph node is negative [ypN0(sn)]; instead, to date, patients with metastatic sentinel lymph node after neoadjuvant chemotherapy, even with only isolated tumor cells, [ypN+(sn) including ypN0i+(sn) and ypN1mic(sn)] should always be treated with AD. However, NAC significantly increases the difficulties and complexity of axillary surgical management. A personalized multidisciplinary path in specialized breast centers should ensure an accurate clinical counselling and refined patient selection for SLNB post-NAC. KEY WORDS: Axillary treatment, Breast cancer, Neoadjuvant chemotherapy, Sentinel lymph node biopsy.


The influence of renal function on diabetic foot ulceration.
We examined the effect of renal function on the formation, severity, and outcome of diabetic foot lesions. Information was collected from a retrospective hospital chart survey and analyzed by univariate and multivariate linear regression analysis. Creatinine clearance, peripheral neuropathy, and peripheral vascular disease were all found to be independently associated with formation of foot lesions, indicating that each of these acts by distinct biologic mechanisms. Renal function had no bearing on the severity of lesions or on their eventual healing. We conclude that foot ulcers are more likely to develop in diabetic patients who also suffer from renal impairment, but they are no less likely to heal than are those in patients with normal renal function. We further conclude that attempts to preserve functional limbs in these patients are justified.


AN IN-DEPTH ASSESSMENT OF THE TUMOR'S IMPACT ON SARCOPENIA.
Loss of muscle mass is a typical symptom of cancer and it is strongly correlated with poor prognosis. Cancer-related Sarcopenia is unresponsive to conventional dietary changes and exercise, in contrast to age-associated muscle atrophy. This particular type of weakness differs from different kinds of muscle loss in that it is triggered by a number of interrelated mechanisms, notably inflammatory processes, abnormal metabolic processes, proteolysis, and autophagy. This research is to examine evidence supporting the theory that tumors have a causal role in causing muscular atrophy. It seeks to investigate the precise regulators that the tumour generates and how they affect the processes that result in muscle waste. The evaluation looks for new directions for further studies and medical treatments. The analysis is based on a thorough examination of the scientific literature and research that shows how tumor and muscle atrophy are related. It concentrates on studies that clarify the numerous strategies by which malignancies cause the loss of muscle. This article highlights particular mechanisms by which these tumor-derived substances affect the development of muscle loss, including inflammatory processes, metabolic disturbance, proteolysis, and autophagy. The discovery of such targets offers hope for the creation of efficient treatment strategies that can enhance the long-term outlook and quality of life of cancer sufferers who are experiencing muscle loss.


A happy story: Developmental changes in children's sensitivity to facial expressions of varying intensities.
Using 20 levels of intensity, we measured children's thresholds to discriminate the six basic emotional expressions from neutral and their misidentification rates. Combined with the results of a previous study using the same method (Journal of Experimental Child Psychology, 102 (2009) 503-521), the results indicate that by 5 years of age, children are adult-like, or nearly adult-like, for happy expressions on all measures. Children's sensitivity to other expressions continues to improve between 5 and 10 years of age (e.g., surprise, disgust, fear) or even after 10 years of age (e.g., anger, sad). The results indicate that there is a slow development of sensitivity to the expression of all basic emotions except happy. This slow development may impact children's social and cognitive development by limiting their sensitivity to subtle expressions of disapproval or disappointment.


[The effect of secondary Bjerknes force in sonoporation on cell by ultrasound-mediated microbubbles].
In sonoporation, the cell membrane is broken-up temporarily by ultrasound mediated microbubbles, which is promoting drug or gene into the cell. In current literatures, there are numerous studies of single microbubble dynamics in sonoporation. However till now, little studies have been focused on the sonoporation incidence caused by more than one microbubble. In this article, the dynamic model of two adjacent microbubbles in stable cavitation has been introduced. By the model, the forces including secondary Bjerknes force on cell membrane given by microbubbles and their effects on sonoporation have been numerically studied. According to the experimental parameters, we numerically studied (1) effects of the ultrasound and microbubble parameters on the secondary Bjerknes forces; (2) the forces exerted on cell membrane by microbubble, including the secondary Bjerknes force; (3) the sonoporation possibility caused by those forces produced by microbubble. In this article, the ultrasound and microbubbles' parameters range were found to produce sonoporation by two adjacent microbubbles. Furthermore, it is the first time to found that the microbubbles' parameters are more important than ultrasound parameters on sonoporation.


Evaluation of oral problems in divers performing electrical welding and cutting under water.
A study of 118 professional commercial divers, using an inquiry form, revealed that 55% of the divers complained about a metallic taste in the mouth when working with electrical equipment under water. A nationwide clinical examination of 40 divers was performed to see if the metallic taste during work reflected any alteration in the oral cavity. The clinical appearance of the dental amalgam restorations differed significantly between divers who had been working with electrical equipment and divers who had not. A number of other dental indexes were assessed and showed no differences between the two groups. These included salivary secretion rate, buffering capacity, secondary caries, plaque index, bleeding index, and cultures of S. mutans and Lactobacillus. Relatively high levels of the two organisms were noted, indicating that the group of divers examined should be considered as a caries risk groups.


Characterization of the yam tuber storage proteins from Dioscorea batatas exhibiting unique lectin activities.
Four major proteins designated DB1, DB2, DB3, and DB4 were isolated and characterized from the yam tuber Dioscorea batatas. The ratios of their yields were 20:50:20:10. DB1 was a mannose-binding lectin (20 kDa) consisting of 10-kDa subunits and was classified as the monocot mannose-binding lectin family. DB2, accounting for 50% of the total protein, was the storage protein, commonly called dioscorins consisting of a 31-kDa subunit. On the basis of amino acid sequence, DB2 was classified to be dioscorin A. DB3 was a maltose-binding lectin, having an apparent molecular mass of 120 kDa and composed of a 66-kDa subunit and two 31-kDa subunits (DB3S). The 66-kDa subunit was further composed of two 31-kDa subunits (DB3L) cross-linked by disulfide bonds. DB3L and DB3S (242 and 241 amino acid residues, respectively) were homologous with each other with 72% sequence identity. They showed a sequence homology to dioscorin B and dioscorin A from Dioscorea alata, with 90 and 93% identity, respectively, and to carbonic anhydrase from Arabidopsis thaliana with about 45% identity. DB3S had one intrachain disulfide bond located at Cys(28)-Cys(187), whereas DB3L had one interchain disulfide bond (Cys(40)-Cys(40)') in addition to the intrachain disulfide bond (Cys(28)-Cys(188)) to form a 66-kDa subunit. DB1 and DB3 agglutinated rabbit erythrocytes at 2.7 and 3.9 microg/ml, respectively. Despite the structural homology between DB2 and DB3, DB2 had no lectin activity. The 66-kDa subunit itself revealed the full hemagglutinating activity of DB3, indicating that DB3L but not DB3S was responsible for the activity. The hemagglutinating activity of DB3 required Ca(2+) ions and was exclusively inhibited by maltose and oligomaltoses (e.g. maltopentaose and maltohexaose) but not by d-glucose. DB3 could not be classified into any known plant lectin family. DB4 was a chitinase, homologous to an acidic chitinase from Dioscorea japonica. DB1, DB2, and DB3 did not show any activity of carbonic anhydrase, amylase, or trypsin inhibitor activity. These results show that two of the four major proteins isolated from the yam tubers D. batatas have unique lectin activities.


Foeniculum vulgare seed extract induces apoptosis in lung cancer cells partly through the down-regulation of Bcl-2.
The factors behind the pathogenesis of lung cancer are not clear, and treatment failure is generally caused by drug resistance, recurrence, and metastasis. Development of new therapeutic agents to overcome drug-resistance remains a challenge clinically. Various extracts of Foeniculum vulgare have shown promising anticancer activity; however, effects on lung cancer and the underlying molecular mechanisms of action are not clear. In the present study, we found that the ethanol extract of Foeniculum vulgare seeds (EEFS) significantly reduced lung cancer cell growth in vitro and in vivo. EEFS decreased the viability of and triggered apoptosis in the lung cancer cell lines NCI-H446 and NCI-H661. EEFS induced apoptosis mainly through inhibition of Bcl-2 protein expression, reduction of mitochondrial membrane potential, and release of Cytochrome C. Moreover, EEFS significantly inhibited colony formation and cell migration in lung cancer cells. EEFS also effectively inhibited the growth of xenograft tumors derived from NCI-446 cells by reducing Bcl-2 protein expression and inducing apoptosis. Taken together, these findings suggest that EEFS exerts anti-lung cancer activity by targeting the Bcl-2 protein and may have potential as a therapeutic drug for lung cancer.


Drug-induced arrhythmias.
The objective of this review is to characterize the mechanisms, risk factors, and offending pharmacotherapeutic agents that may cause drug-induced arrhythmias in critically ill patients. PubMed, other databases, and citation review were used to identify relevant published literature. The authors independently selected studies based on relevance to the topic. Numerous drugs have the potential to cause drug-induced arrhythmias. Drugs commonly administered to critically ill patients are capable of precipitating arrhythmias and include antiarrhythmics, antianginals, antiemetics, gastrointestinal stimulants, antibacterials, narcotics, antipsychotics, inotropes, digoxin, anesthetic agents, bronchodilators, and drugs that cause electrolyte imbalances and bradyarrhythmias. Drug-induced arrhythmias are insidious but prevalent. Critically ill patients frequently experience drug-induced arrhythmias; however, enhanced appreciation for this adverse event has the potential to improve prevention, treatment, patient safety, and outcomes in this patient population.


A modified hMG-GnRH method for the induction of ovulation in infertile women with severe hypogonadotropic amenorrhea.
The objective of this study was to compare, in infertile women suffering from severe hypogonadotropic amenorrhea, the therapeutic utility and the incidence of complications arising from fertility treatment by the conventional human menopausal gonadotropin/human chorionic gonadotropin (hMG-hCG) method, the hMG step-down method, the sequential hMG/gonadotropin-releasing hormone (GnRH) method and a new, modified hMG-GnRH method that has been developed by us. In the step-down method, the daily dose of hMG was decreased from 150 IU to 75 IU when the follicle diameter reached 11-13 mm. In the sequential hMG-GnRH, hMG injection was switched to pulsatile GnRH administration (20 microg/120 min SC), when the follicle diameter reached 11-13 mm. In our new modified hMG-GnRH, pulsatile GnRH was injected together with hMG. Daily hMG was stopped and the GnRH dosage was changed from 10 microg to 20 microg when the follicle diameter reached 11-13 mm. Initially, the three established methods were applied randomly to treat 34 cycles in 20 women; and subsequently, five patients who failed to conceive following treatment by sequential hMG-GnRH were then treated by the modified hMG-GnRH method. More than eight growing follicles and multiple pregnancies were observed during treatment by the conventional method. The incidence of ovarian hyperstimulation syndrome (OHSS) was 25.7% with the conventional method, 20.0% with the step-down method and 0% with the sequential hMG-GnRH method; however, the rate of ovulation was only 50% with the sequential hMG-GnRH method. By contrast, with the modified hMG-GnRH method, less than three growing follicles occurred in 81.8% of patients, there was a 100% rate of ovulation, and neither OHSS nor multiple pregnancies were observed. Moreover, the modified hMG-GnRH method induced pregnancy in 3 out of 5 patients. These data indicate that this new method is favorable for the treatment of severe hypogonadotropic amenorrhea.


Molecular and serological surveillance of canine enteric viruses in stray dogs from Vila do Maio, Cape Verde.
Infections caused by canine parvovirus, canine distemper virus and canine coronavirus are an important cause of mortality and morbidity in dogs worldwide. Prior to this study, no information was available concerning the incidence and prevalence of these viruses in Cape Verde archipelago. To provide information regarding the health status of the canine population in Vila do Maio, Maio Island, Cape Verde, 53 rectal swabs were collected from 53 stray dogs during 2010 and 93 rectal swabs and 88 blood samples were collected from 125 stray dogs in 2011. All rectal swabs (2010 n = 53; 2011 n = 93) were analysed for the presence of canine parvovirus, canine distemper virus and canine coronavirus nucleic acids by quantitative PCR methods. Specific antibodies against canine distemper virus and canine parvovirus were also assessed (2011 n = 88).From the 2010 sampling, 43.3% (23/53) were positive for canine parvovirus DNA, 11.3% (6/53) for canine distemper virus RNA and 1.9% (1/53) for canine coronavirus RNA. In 2011, the prevalence values for canine parvovirus and canine coronavirus were quite similar to those from the previous year, respectively 44.1% (41/93), and 1.1% (1/93), but canine distemper virus was not detected in any of the samples analysed (0%, 0/93). Antibodies against canine parvovirus were detected in 71.6% (63/88) blood samples and the seroprevalence found for canine distemper virus was 51.1% (45/88). This study discloses the data obtained in a molecular and serological epidemiological surveillance carried out in urban populations of stray and domestic animals. Virus transmission and spreading occurs easily in large dog populations leading to high mortality rates particularly in unvaccinated susceptible animals. In addition, these animals can act as disease reservoirs for wild animal populations by occasional contact. Identification of susceptible wildlife of Maio Island is of upmost importance to evaluate the risk of pathogen spill over from domestic to wild animals in Cape Verde and to evaluate the associated threat to the wild susceptible species.


Metabolite Signature of Life's Essential 8 and Risk of Coronary Heart Disease Among Low-Income Black and White Americans.
Life's essential 8 (LE8) is a comprehensive construct of cardiovascular health. Yet, little is known about the LE8 score, its metabolic correlates, and its predictive implications among Black Americans and low-income individuals. In a nested case-control study of coronary heart disease (CHD) among 299 pairs of Black and 298 pairs of White low-income Americans from the Southern Community Cohort Study, we estimated LE8 score and applied untargeted plasma metabolomics and elastic net with leave-one-out cross-validation to identify metabolite signature (MetaSig) of LE8. Associations of LE8 score and MetaSig with incident CHD were examined using conditional logistic regression. The mediation effect of MetaSig on the LE8-CHD association was also examined. The external validity of MetaSig was evaluated in another nested CHD case-control study among 299 pairs of Chinese adults. Higher LE8 score was associated with lower CHD risk (standardized odds ratio, 0.61 [95% CI, 0.53-0.69]). The MetaSig, consisting of 133 metabolites, showed significant correlation with LE8 score (r=0.61) and inverse association with CHD (odds ratio, 0.57 [0.49-0.65]), robust to adjustment for LE8 score and across participants with different sociodemographic and health status ([odds ratios, 0.42-0.69]; all P<0.05). MetaSig mediated a large portion of the LE8-CHD association: 53% (32%-80%). Significant associations of MetaSig with LE8 score and CHD risk were found in validation cohort (r=0.49; odds ratio, 0.57 [0.46-0.69]). Higher LE8 score and its MetaSig were associated with lower CHD risk among low-income Black and White Americans. Metabolomics may offer an objective measure of LE8 and its metabolic phenotype relevant to CHD prevention among diverse populations.


Simple, almost anywhere, with almost anyone: remote low-cost telementored resuscitative lung ultrasound.
Apnea (APN) and pneumothorax (PTX) are common immediately life-threatening conditions. Ultrasound is a portable tool that captures anatomy and physiology as digital information allowing it to be readily transferred by electronic means. Both APN and PTX are simply ruled out by visualizing respiratory motion at the visceral-parietal pleural interface known as lung sliding (LS), corroborated by either the M-mode or color-power Doppler depiction of LS. We thus assessed how economically and practically this information could be obtained remotely over a cellular network. Ultrasound images were obtained on handheld ultrasound machines streamed to a standard free internet service (Skype) using an iPhone. Remote expert sonographers directed remote providers (with variable to no ultrasound experience) to obtain images by viewing the transmitted ultrasound signal and by viewing the remote examiner over a head-mounted webcam. Examinations were conducted between a series of remote sites and a base station. Remote sites included two remote on-mountain sites, a small airplane in flight, and a Calgary household, with base sites located in Pisa, Rome, Philadelphia, and Calgary. In all lung fields (20/20) on all occasions, LS could easily and quickly be seen. LS was easily corroborated and documented through capture of color-power Doppler and M-mode images. Other ultrasound applications such as the Focused Assessment with Sonography for Trauma examination, vascular anatomy, and a fetal wellness assessment were also demonstrated. The emergent exclusion of APN-PTX can be immediately accomplished by a remote expert economically linked to almost any responder over cellular networks. Further work should explore the range of other physiologic functions and anatomy that could be so remotely assessed.


Muddied Waters: The Use of "Residual" And "Legacy" Phosphorus.
Phosphorus (P) inputs to the biosphere have quadrupled in less than a century due to intensification of rock phosphate mining and the use of P fertilizers for crop production. Accumulation of P in soils can increase P transfers across the soil-water continuum that impair aquatic ecosystem function and water resource quality for society. However, what this accumulated P is called, and subsequent connotations of magnitude versus mechanism at pedon versus watershed scale, varies in the literature. We argue that the two commonly used terms of "residual" and "legacy" P, though often used interchangeably, hold distinct meanings and connotations. Tracing the historical origins and trajectories of these terms reveals that "residual P" refers to the magnitude of fertilizer P that remains in the soil after crop harvest, whereas "legacy P" refers to the mechanism of P transfer across the watershed and its long-term impacts on water quality. The use of "legacy P" in many cases refers to the residuality of anthropogenic P inputs, and thus should be "residual P". We recommend that the term "residual P" be used when referring to the accumulation of P in soils under agricultural management from past inputs, and the term "legacy P" be used when referring to the transfer of P within watersheds. The intentional and thus consistent use of residual versus legacy P stands to provide important nuance in the environmental sciences and overlapping fields of agronomy and biogeochemistry.


Anticipatory nausea and vomiting.
A commonly reported consequence of post-treatment nausea or vomiting is the development of anticipatory nausea and vomiting (ANV). In most published work, nausea is reported to occur before chemotherapy drugs are administered by approximately 20% of patients at any one chemotherapy cycle and by 25-30% of patients by their fourth chemotherapy cycle. Most studies in adult patients strongly support the view that the development of ANV involves elements of classical conditioning. The best method to avoid development of ANV is to adequately prevent both vomiting and nausea from the first exposure to chemotherapy. If anticipatory side effects develop, behavioral treatment techniques, such as systematic desensitization, have been shown effective. Benzodiazepines used in combination with behavioral techniques or antiemetics may also be useful. The evidence on which these conclusions are based is reviewed in this article.


Rapid identification of tick-borne encephalitis virus by the fluorescent antibody technique.
An immunofluorescence technique in tissue cultures and in mice, developed to provide a rapid means of identification of group B arboviruses, has been used to identify the tick-borne encephalitis virus. Identification in mouse-brain tissue was not possible when mice with vague clinical symptoms were used. Identification in chick-embryo-cell cultures was less time-consuming and laborious than identification in mouse brain, but more care was required, since only a small percentage of cells showed fluorescence. Identification can be accomplished in a few hours if mice in agony or cell cultures three to five days after inoculation are used.


The Hidden Dimension: Context-Dependent Expression of Repeatable Behavior in Copepods.
In ecotoxicology and aquatic ecology, we often ignore responses of individuals and focus on average responses. However, both terrestrial and aquatic animals display consistent behavioral differences between individuals. The distribution of behavioral differences within a population contains vital information for predicting population responses to novel environmental challenges. Currently, individual data for behavioral and physiological traits of small marine invertebrates are few, partly because such variation is lost within published group means and assumed normality. We tested the combined effects of an inorganic contaminant (copper) and a biological stressor (i.e., chemical cues of a fish predator) on activity in a marine copepod. Although direct stress effects were weak, individuals behaved consistently differently, depending on the context. Individual differences in behavior were only expressed under the influence of kairomones, but not by copper exposure alone. This finding indicates that copepods express repeatable and context-dependent behavior. We also demonstrate how large variations in behavioral data can hide consistent differences between individuals. Environ Toxicol Chem 2020;39:1017-1026. © 2020 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.


A prospective, randomized, double-blind, multicenter study comparing remifentanil with fentanyl in mechanically ventilated patients.
To compare the quality of analgesia provided by a remifentanil-based analgesia regime with that provided by a fentanyl-based regime in critically ill patients. This was a registered, prospective, two-center, randomized, triple-blind study involving adult medical and surgical patients requiring mechanical ventilation (MV) for more than 24 h. Patients were randomized to either remifentanil infusion or a fentanyl infusion for a maximum of 30 days. Sedation was provided using propofol (and/or midazolam if required). Primary outcome was the proportion of patients in each group maintaining a target analgesia score at all time points. Secondary outcomes included duration of MV, discharge times, and morbidity. At planned interim analysis (n = 60), 50% of remifentanil patients (n = 28) and 63% of fentanyl patients (n = 32) had maintained target analgesia scores at all time points (p = 0.44). There were no significant differences between the groups with respect to mean duration of ventilation (135 vs. 165 h, p = 0.80), duration of hospital stay, morbidity, or weaning. Interim analysis strongly suggested futility and the trial was stopped. The use of remifentanil-based analgesia in critically ill patients was not superior regarding the achievement and maintenance of sufficient analgesia compared with fentanyl-based analgesia.


Genetic analysis of murine hepatitis virus strain JHM.
We performed a genetic analysis of 37 temperature-sensitive mutants of murine hepatitis virus strain JHM. Of our mutants, 32 did not induce murine hepatitis virus-specific RNA synthesis in infected cells at the restrictive temperature, 39 degrees C. By complementation testing we have identified at least seven nonoverlapping complementation groups. Six of the genes identified in this way are required for murine hepatitis virus-specific RNA synthesis. The seventh complementation group is made up of five mutants which induced virus-specific RNA synthesis at 39 degrees C.


Palliative combined cutting and high-pressure balloon valvuloplasty in six dogs with severe, symptomatic subaortic stenosis.
Severe subaortic stenosis (SAS) is a congenital heart defect in dogs that often results in clinical signs and reduced survival. The objective of this study was to describe characteristics of dogs with severe, symptomatic SAS who underwent combined cutting and high-pressure balloon valvuloplasty (CB/HPBV). Retrospective description of the clinical characteristics, CB/HPBV procedural deviations from reported methodology and outcomes in a series of six client-owned dogs with severe, symptomatic SAS. Breeds included two each of Newfoundland, Golden retriever, and German shepherd. Median age was 10.1 months (range: 5-72.3 months), and median weight was 25.5 kg (range: 21.8-36.4 kg). Before CB/HPBV, clinical signs were present in all dogs; four were managed for congestive heart failure (CHF). Three dogs had concurrent congenital heart disease. Median Doppler-estimated left ventricular outflow tract pressure gradient was pre-operatively 149.7 mmHg (range: 89.9-254.7 mmHg) and post-operatively 134.1 mmHg (range: 83.9-181.2 mmHg). Median aortoseptal angle was steep at 136° (range: 109-143°). Clinical improvement was documented in all dogs, based on temporary discontinuation of diuretics and/or owner-perceived reduction in clinical signs. At the time of writing, three dogs had died suddenly, one was euthanized because of recurrence of clinical signs, and one died in CHF. Median survival time was 26.4 months after procedure (range: 6.3-45.8 months). One dog remained alive at 44 months after procedure. Palliative CB/HPBV is a potential therapeutic option for dogs with severe, symptomatic SAS complicated by concurrent congenital heart disease, arrhythmias, or CHF.


Confusion assessment method to analyze delirium in intensive care unit: literature review.
Delirium is frequently observed in intensive care unit patients and its occurrence is related to increased morbidity and mortality, length of stay, functional decline and high costs. The Confusion Assessment Method for Intensive Care Unit is a tool that facilitates early identification and occurrence of delirium among intubated patients. To verify the aspects of delirium studied by means of the Confusion Assessment Method for Intensive Care Unit. A literature review was conducted in the LILACS, MedLine, PubMed and CINAHL databases, from 2001, when the Confusion Assessment Method for Intensive Care Unit was validated, to 2008. Descriptors used for LILACS, MedLine and PubMed were delirium and intensive care unit, while for the CINAHL database, delirium and intensive care were used. From 293 articles, 35 were selected. The aspects analyzed disclosed, different types of delirium in different intensive care units. Variation in sensitivity was of 93% to 100% and variation in specificity, 89% to 100% of the Confusion Assessment Method for Intensive Care Unit, an important tool for detection, characterization and control of delirium and its impact. The aspects of delirium studied by means of the Confusion Assessment Method for Intensive Care Unit were: the performance index, identification, management, cost of treatment, morbidity and mortality due to delirium.


An Expanded Conceptual Framework of Medical Students' Primary Care Career Choice.
In many countries, the number of graduating medical students pursuing a primary care career does not meet demand. These countries face primary care physician shortages. Students' career choices have been widely studied, yet many aspects of this process remain unclear. Conceptual models are useful to plan research and educational interventions in such complex systems.The authors developed a framework of primary care career choice in undergraduate medical education, which expands on previously published models. They used a group-based, iterative approach to find the best way to represent the vast array of influences identified in previous studies, including in a recent systematic review of the literature on interventions to increase the proportion of students choosing a primary care career. In their framework, students enter medical school with their personal characteristics and initial interest in primary care. They complete a process of career decision making, which is subject to multiple interacting influences, both within and outside medical school, throughout their medical education. These influences are stratified into four systems-microsystem, mesosystem, exosystem, and macrosystem-which represent different levels of interaction with students' career choices.This expanded framework provides an updated model to help understand the multiple factors that influence medical students' career choices. It offers a guide for the development of new interventions to increase the proportion of students choosing primary care careers and for further research to better understand the variety of processes involved in this decision.


Antigen-induced arthritis: inflammation and antigen handling after two different doses of intra-articularly injected antigen.
We studied (i) the inflammatory response induced by intra-articular (i.a.) injection of 2.5 and 0.5 mg 125I-labelled bovine serum albumin (125I-BSA) into the paired knee joints of rabbits immunized with BSA in Freund's complete adjuvant, and (ii) handling of these different doses of antigen by means of regular external radioactivity measurements. The joint inflammation was quantified by 99mTechnetium pertechnetate uptake measurements. More severe arthritis was seen in the 2.5 mg BSA-injected than in the paired 0.5 mg BSA-injected knee, both in the early and in the late phase of antigen-induced arthritis. External radioactivity measurements showed enhanced disappearance of radioactivity from the 2.5 mg 125I-BSA challenged knee joint, resulting in lower percentual retention of the dose injected than in the 0.5 mg injected knee joint. However the calculated absolute amount of long term retained BSA in the 2.5 mg BSA injected knee, 4 weeks after i.a. injection, was still about 2.8 times the quantity of BSA retained in the 0.5 mg BSA-injected paired knee joint. These data indicate that the severity of both the early and late phase of antigen-induced arthritis is i.a. antigen dose-dependent and, in addition, suggest that handling of i.a. antigen depends in part on the severity of joint inflammation.


Isolation, purification, and chemical analysis of the lipopolysaccharide and lipid A of Acinetobacter calcoaceticus NCTC 10305.
The lipopolysaccharide of Acinetobacter calcoaceticus NCTC 10305 (London) was obtained by a modified phenol/chloroform/light petroleum method from the bacterial cells and from the culture medium in yields of 1.6% and 2.2% respectively (based on the bacterial dry weight). On chemical analysis, both preparations proved to be identical. The lipopolysaccharide obtained from the cells was purified by repeated ultracentrifugation, electrodialysis, and precipitation with sodium chloride. It was free of nucleic acids, proteins, and glycans. In the analytical ultracentrifuge, the triethylamine and sodium salt forms of the lipopolysaccharide showed a s20 value of 8.9 S and 51 S, respectively. The lipopolysaccharide consisted of glucosamine, 3-deoxy-D-manno-octulosonic acid, D-glucose, fatty acids, and phosphate in a molar ratio of 2:1:7:6:4. The fatty acids were predominantly lauric acid, 2-hydroxy, and 3-hydroxylauric acid in a molar ratio of 1:1:2. Only 3-hydroxylauric acid was found in amide linkage. On mild acid hydrolysis of the lipopolysaccharide, 65% lipid A were obtained, to which glucosamine was retained quantitatively. It still contained 50% of the original glucose, while one third (15%) of the liberated glucose was in monomeric form.


Pigment genes not skin pigmentation affect UVB-induced vitamin D.
Skin pigmentation is believed to contribute to the generally low serum 25-hydroxyvitamin D (25(OH)D) concentrations observed in darker-skinned persons. The influence of measured skin pigmentation on UVB-induced 25(OH)D increase was investigated together with 9 demographic and 13 genetic parameters (pigment SNPs). Forty participants representing a wide range in measured skin pigmentation were exposed to identical UVB doses on identical body areas over nine weeks with weekly measurements of serum 25(OH)D. This study took place in Denmark during winter, a period with negligible ambient UVB, so variation in 25(OH)D synthesis was not influenced by latitude, season, sun and clothing habits. The increase in 25(OH)D concentration displayed considerable variation (range: 2.9 to 139 nmol L-1). Constitutive and facultative skin pigmentation exerted separate influence on the variation of the UVB-induced linear 25(OH)D increase. However, this influence was statistically non-significant in the presence of separate significant pigment SNPs. The variation in the 25(OH)D increase in the combined linear model was not explained by measured skin pigmentation but by sex, height, age and seven SNPs located in the ASIP, MTAP, MIR196A29 and Solute Carrier Family genes. This linear model including individual intercepts and the 10 parameters influencing the slope explained 77.4% of the variation. This study confirmed the influence of sex, age and height on 25(OH)D increase and found that pigment genes provided a better relation to UVB-induced 25(OH)D increase compared to the actual measured skin pigmentation. Therefore, only investigating skin pigmentation obscures other causal parameters for low 25(OH)D.


Progress toward Room-Temperature Synthesis and Functionalization of Iron-Oxide Nanoparticles.
Novel magnetic nanohybrids composed of nanomaghemite covered by organic molecules were successfully synthesized at room temperature with different functionalization agents (sodium polystyrene sulfonate, oxalic acid, and cetyltrimethylammonium bromide) in low and high concentrations. Structural, vibrational, morphological, electron energy-loss spectroscopy, magnetic, and Mössbauer characterizations unraveled the presence of mainly cubic inverse spinel maghemite (γ-Fe2O3), whilst X-ray diffraction and 57Fe Mössbauer spectroscopy showed that most samples contain a minor amount of goethite phase (α-FeOOH). Raman analysis at different laser power revealed a threshold value of 0.83 mW for all samples, for which the γ-Fe2O3 to α-Fe2O3 phase transition was observed. Imaging microscopy revealed controlled-size morphologies of nanoparticles, with sizes in the range from 8 to 12 nm. Organic functionalization of the magnetic nanoparticles was demonstrated by vibrational and thermogravimetric measurements. For some samples, Raman, magnetic, and Mössbauer measurements suggested an even more complex core-shell-like configuration, with a thin shell containing magnetite (Fe3O4) covering the γ-Fe2O3 surface, thus causing an increase in the saturation magnetization of approximately 11% against nanomaghemite. Field cooling hysteresis curves at 5 K did not evidence an exchange bias effect, confirming that the goethite phase is not directly interacting magnetically with the functionalized maghemite nanoparticles. These magnetic nanohybrids may be suitable for applications in effluent remediation and biomedicine.


Prevalence, severity, and relationships of lung lesions, liver abnormalities, and rumen health scores measured at slaughter in beef cattle.
An array of management tools exists within the beef industry to improve animal welfare and productivity; however, the ability to assess the outcomes of these tools is needed. Deficiencies in management commonly manifest as bovine respiratory disease complex or nutritional disorders such as acidosis; therefore, lung, liver, and rumen gross pathology lesions present at slaughter were measured as part of the Harvest Audit Program (HAP) and associations with performance determined. Individual gross pathology data from 19,229 cattle at commercial packing plants in Kansas and Texas were collected. Corresponding individual preharvest and carcass data were obtained on a subset of 13,226 cattle. Associations between lesions and performance were modeled using multivariable mixed effect models. Regression coefficients were used for estimation of lesion associative effects on continuous outcomes and odds ratios for dichotomous outcomes. Across the entire population, 67.3% of the cattle had no pulmonary lesions; 22.5 and 9.8% of cattle displayed mild and severe lesions, respectively. Severe pulmonary lesions were associated with a decreased ADG of 0.07 kg and a HCW 7.1 kg less than cohorts with no pulmonary lesions (P < 0.01). Overall, 68.6% of cattle observed had normal livers. Of cattle severely affected by liver abscesses (A+; 4.6%), 14.9% also displayed severe pulmonary lesions and 28.3% displayed mild pulmonary lesions. Rumenitis lesions were observed in 24.1% of the overall study population. Of cattle with mildly abscessed livers (A-), moderately abscessed livers (A), and severely abscessed livers, 20.6, 21.6, and 9.24% displayed mild or severe rumenitis lesions at slaughter. Severe rumenitis lesions were associated with a significant decrease in ADG and HCW (0.025 and 2.20 kg, respectively; P < 0.001). Although the majority of the cattle in this population would be considered low risk, after adjustments for cattle with multiple lesions, 22.9% of cattle in the overall population were observed with a severe lesion (lung, liver, or rumen). In conclusion, a gross pathology monitoring system is feasible and the 22.9% prevalence of severe lesions (lung, liver, or rumen) indicates that significant opportunity exists to improve beef cattle health, well-being, and productivity. Data generated using HAP may be used to support decisions concerning the implementation or removal of managerial practices and health interventions in beef cattle production systems.


Population pharmacokinetics and pharmacodynamic modeling of abacavir (1592U89) from a dose-ranging, double-blind, randomized monotherapy trial with human immunodeficiency virus-infected subjects.
Abacavir (formerly 1592U89) is a carbocyclic nucleoside analog with potent anti-human immunodeficiency virus (anti-HIV) activity when administered alone or in combination with other antiretroviral agents. The population pharmacokinetics and pharmacodynamics of abacavir were investigated in 41 HIV type 1 (HIV-1)-infected, antiretroviral naive adults with baseline CD4(+) cell counts of >/=100/mm(3) and plasma HIV-1 RNA levels of >30,000 copies/ml. Data for analysis were obtained from patients who received randomized, blinded monotherapy with abacavir at 100, 300, or 600 mg twice-daily (BID) for up to 12 weeks. Plasma abacavir concentrations from sparse sampling were analyzed by standard population pharmacokinetic methods, and the effects of dose, combination therapy, gender, weight, and age on parameter estimates were investigated. Bayesian pharmacokinetic parameter estimates were calculated to determine the peak concentration of abacavir in plasma (C(max)) and the area under the concentration-time curve from time zero to infinity (AUC(0-infinity)) for individual subjects. The pharmacokinetics of abacavir were dose proportional over the 100- to 600-mg dose range and were unaffected by any covariates. No significant correlations were observed between the incidence of the five most common adverse events (headache, nausea, diarrhea, vomiting, and malaise or fatigue) and AUC(0-infinity). A significant correlation was observed between C(max) and nausea by categorical analysis (P = 0.019), but this was of borderline significance by logistic regression (odds ratio, 1.45; 95% confidence interval, 0.95 to 2.32). The log(10) time-averaged AUC(0-infinity) minus baseline (AAUCMB) values for HIV-1 RNA and CD4(+) cell count correlated significantly with C(max) and AUC(0-infinity), but with better model fits for AUC(0-infinity). The increase in AAUCMB values for CD4(+) cell count plateaued early for drug exposures that were associated with little change in AAUCMB values for plasma HIV-1 RNA. There was less than a 0.4 log(10) difference over 12 weeks in the HIV-1 RNA levels with the doubling of the abacavir AUC(0-infinity) from 300 to 600 mg BID dosing. In conclusion, pharmacodynamic modeling supports the selection of abacavir 300 mg twice-daily dosing.


Azithromycin vs cefuroxime plus erythromycin for empirical treatment of community-acquired pneumonia in hospitalized patients: a prospective, randomized, multicenter trial.
To compare the efficacy and safety of azithromycin dihydrate monotherapy with those of a combination of cefuroxime axetil plus erythromycin as empirical therapy for community-acquired pneumonia in hospitalized patients. Patients were enrolled in a prospective, randomized, multicenter study. The standard therapy of cefuroxime plus erythromycin was consistent with the American Thoracic Society, Canadian Community-Acquired Pneumonia Consensus Group, and Infectious Disease Society of America consensus guidelines. The doses were intravenous azithromycin (500 mg once daily) followed by oral azithromycin (500 mg once daily), intravenous cefuroxime (750 mg every 8 hours), followed by oral cefuroxime axetil (500 mg twice daily), and erythromycin (500-1000 mg) intravenously or orally every 6 hours. Randomization was stratified by severity of illness and age. Patients who were immunosuppressed or residing in nursing homes were excluded. Data from 145 patients (67 received azithromycin and 78 received cefuroxime plus erythromycin) were evaluable. Streptococcus pneumoniae and Haemophilus influenzae were isolated in 19% (28/145) and 13% (19/145), respectively. The atypical pathogens accounted for 33% (48/145) of the etiologic diagnoses; Legionella pneumophila, Chlamydia pneumoniae, and Mycoplasma pneumoniae were identified in 14% (20/ 145), 10% (15/145), and 9% (13/145), respectively. Clinical cure was achieved in 91% (61/67) of the patients in the azithromycin group and 91% (71/78) in the cefuroxime plus erythromycin group. Adverse events (intravenous catheter site reactions, gastrointestinal tract disturbances) were significantly more common in patients who received cefuroxime plus erythromycin (49% [30/78]) than in patients who received azithromycin (12% [8/67]) (P<.001). Treatment with azithromycin was as effective as cefuroxime plus erythromycin in the empirical management of community-acquired pneumonia in immunocompetent patients who were hospitalized. Azithromycin was well tolerated.


Surgical outcome and prognostic factors of anterior decompression and fusion for cervical compressive myelopathy due to ossification of the posterior longitudinal ligament.
Anterior decompression and fusion (ADF) for ossification of the posterior longitudinal ligament (OPLL) is technically demanding and associated with complications. Although various factors affecting clinical outcome have been investigated in posterior decompression, prognostic factors of ADF remain unclear. The purpose of the study was to identify surgical outcome and prognostic factors of ADF for cervical myelopathy due to OPLL. This was a retrospective case study. Between 2005 and 2012, 913 patients underwent decompression surgery for cervical OPLL at our institution. Among them, 131 who underwent ADF and 221 who underwent laminoplasty were enrolled. Inclusion criteria were (1) diagnosis of OPLL; (2) cervical compressive myelopathy; and (3) no trauma, infection, tumor, or previous surgery. We excluded 60 patients with ADF and 157 patients with laminoplasty owing to inadequate follow-up or absence of preoperative myelopathy. Finally, 71 patients with ADF and 64 patients with laminoplasty were enrolled in this study (mean follow-up, 48 vs 41 months). Neurologic assessment was conducted using the Japanese Orthopedic Association (JOA) scoring system for cervical myelopathy. Rate of neurologic improvement was calculated by comparing preoperative and postoperative JOA scores. We investigated the effects of such variables as age, gender, body mass index (BMI), presence of diabetes mellitus (DM), smoking history, type of OPLL, shape of the ossified lesion, occupying ratio of OPLL, presence of intramedullary increased signal intensity (ISI) on magnetic resonance imaging (MRI), and sagittal alignment of the cervical spine on surgical outcome. Severity of ISI was classified into three groups based on T2-weighted sagittal MRI as follows: Grade 0, none; Grade 1, ISI limited to one disc level; or Grade 2, ISI beyond one disc level. This work was supported by the 2013 Korea Health Technology R&D Project of the Ministry of Health and Welfare of the Republic of Korea (A120254). In patients with an occupying ratio ≥60% or with presence of ISI on MRI, ADF yielded better surgical outcome than laminoplasty. A higher ISI grade (B=-28.5, p=.000) and a higher occupying ratio (B=0.88, p=.04) were significantly associated with a lower recovery rate (R=0.56, p=.006). Older age also was associated with a lower recovery rate. Gender, BMI, presence of DM, smoking history, type of OPLL, shape of the ossified lesion, and cervical alignment were not associated with recovery rate. Anterior decompression and fusion has favorable outcome in patients with an occupying ratio ≥60% or with presence of ISI on MRI. Presence of higher ISI grade, higher occupying ratio, and older age were associated with a poor long-term surgical prognosis. Therefore, evaluating ISI and occupying ratio on preoperative MRI is important for selecting the appropriate surgical approach and for predicting clinical outcome after surgery for cervical compressive myelopathy due to OPLL.


Suicidality in bipolar I disorder.
People with bipolar disorder are at high suicide risk. The literature suggests that suicidality is predicted by higher symptom severity and less use of pharmacological agents, but few studies have examined the joint contributions of these variables. The present study examines the conjoint contribution of symptom severity and pharmacological treatment to suicidal ideation and behavior among participants with bipolar disorder. The model was able to account for 53% of the variance in suicidality scores. Depression, mixed state, and hopelessness were significantly associated with suicidality. All other variables were nonsignificant once symptom severity had been controlled. Implications for future research are described.


Fetal MRI: techniques and protocols.
The development of ultrafast sequences has led to a significant improvement in fetal MRI. Imaging protocols have to be adjusted to the rapidly developing fetal central nervous system (CNS) and to the clinical question. Sequence parameters must be changed to cope with the respective developmental stage, to produce images free from motion artefacts and to provide optimum visualization of the region and focus of interest. In contrast to postnatal studies, every suspect fetal CNS abnormality requires examination of the whole fetus and the extrafetal intrauterine structures including the uterus. This approach covers both aspects of fetal CNS disorders: isolated and complex malformations and cerebral lesions arising from the impaired integrity of the feto-placental unit.


Microenvironmental control of glucose metabolism in tumors by regulation of pyruvate dehydrogenase.
During malignant progression cancer cells undergo a series of changes, which promote their survival, invasiveness and metastatic process. One of them is a change in glucose metabolism. Unlike normal cells, which mostly rely on the tricarboxylic acid cycle (TCA), many cancer types rely on glycolysis. Pyruvate dehydrogenase complex (PDC) is the gatekeeper enzyme between these two pathways and is responsible for converting pyruvate to acetyl-CoA, which can then be processed further in the TCA cycle. Its activity is regulated by PDP (pyruvate dehydrogenase phosphatases) and PDHK (pyruvate dehydrogenase kinases). Pyruvate dehydrogenase kinase exists in 4 tissue specific isoforms (PDHK1-4), the activities of which are regulated by different factors, including hormones, hypoxia and nutrients. PDHK1 and PDHK3 are active in the hypoxic tumor microenvironment and inhibit PDC, resulting in a decrease of mitochondrial function and activation of the glycolytic pathway. High PDHK1/3 expression is associated with worse prognosis in patients, which makes them a promising target for cancer therapy. However, a better understanding of PDC's enzymatic regulation in vivo and of the mechanisms of PDHK-mediated malignant progression is necessary for the design of better PDHK inhibitors and the selection of patients most likely to benefit from such inhibitors.


A Quality Improvement Initiative to Increase Scoring Consistency and Accuracy of the Finnegan Tool: Challenges in Obtaining Reliable Assessments of Drug Withdrawal in Neonatal Abstinence Syndrome.
Current practice for diagnosing neonatal abstinence syndrome and guiding pharmacological management of neonatal drug withdrawal is dependent on nursing assessments and repeated evaluation of clinical signs. This single-center quality improvement initiative was designed to improve accuracy and consistency of Finnegan scores among neonatal nurses. One-hundred seventy neonatal nurses participated in a single-session withdrawal-assessment program that incorporated education, scoring guidelines, and a restructured Finnegan scale. Nurses scored a standardized video-recorded infant presenting with opioid withdrawal before and after training. Nearly twice as many nurses scored at target (Finnegan score of 8) posttraining (34.7%; mean error = 0.559, SD = 1.4) compared with pretraining (18.8%; mean error = 1.31, SD = 1.95; Wilcoxon, P < .001). Finnegan scores were significantly higher than the target score pretraining (mean = 9.31, SD = 1.95) compared with posttraining (mean = 8.56, SD = 1.40, Wilcoxon P < .001); follow-up assessments reverted to pretraining levels (mean = 9.16, SD = 1.8). Score dispersion was greater pretraining (variance 3.80) compared with posttraining (variance 1.96; Kendall's Coefficient, P < .001) largely due to score disparity among central nervous system symptomology. Education, clinical guidelines, and a restructured scoring tool increased consistency and accuracy of infant withdrawal-assessments among neonatal nurses. However, more than 60% of nurses did not assess withdrawal to the target score immediately following the training period and improvements did not persist over time. This study highlights the need for more objective tools to quantify withdrawal severity given that assessments are the primary driver of pharmacological management in neonatal drug withdrawal.Video Abstract available at https://journals.lww.com/advancesinneonatalcare/Pages/videogallery.aspx.


Polymorphisms in drug-metabolizing enzymes and steady-state exemestane concentration in postmenopausal patients with breast cancer.
Discovery of clinical and genetic predictors of exemestane pharmacokinetics was attempted in 246 postmenopausal patients with breast cancer enrolled on a prospective clinical study. A sample was collected 2 h after exemestane dosing at a 1- or 3-month study visit to measure drug concentration. The primary hypothesis was that patients carrying the low-activity CYP3A4*22 (rs35599367) single-nucleotide polymorphism (SNP) would have greater exemestane concentration. Additional SNPs in genes relevant to exemestane metabolism (CYP1A1/2, CYP1B1, CYP3A4, CYP4A11, AKR1C3/4, AKR7A2) were screened in secondary analyses and adjusted for clinical covariates. CYP3A4*22 was associated with a 54% greater exemestane concentration (P<0.01). Concentration was greater in patients who reported White race, had elevated aminotransferases, renal insufficiency, lower body mass index and had not received chemotherapy (all P<0.05), and CYP3A4*22 maintained significance after adjustment for covariates (P<0.01). These genetic and clinical predictors of exemestane concentration may be useful for treatment individualization in patients with breast cancer.


Neuro-COVID-19 is more than anosmia: clinical presentation, neurodiagnostics, therapies, and prognosis.
To provide an overview on current knowledge of neurological symptoms and complications of COVID-19, and to suggest management concepts. Headache, dizziness, excessive tiredness, myalgia, anosmia/hyposmia, and ageusia/dysgeusia are common nonspecific neurological manifestations during early COVID-19 disease found in the majority of patients. Less frequent but more severe and specific neurological manifestations include Guillain--Barré syndrome, encephalopathy, encephalitis/meningitis, epileptic seizures, and cerebrovascular events. Beyond standard neurological examination, these require a more extensive work-up, including cerebrospinal fluid assessment, neurophysiological evaluation, neuroimaging, and cognitive testing. Symptomatic treatment is advisable unless the neurological complication's immune pathogenesis is proven. Neurological manifestations of COVID-19 occur during the acute, para-infectious, and 'recovery' phase. Therapeutic management depends on the clinical presentation and neurological work-up.


Acoustic parameters in human speaker recognition.
Four speaker identification tests were conducted using five female speakers known to the listeners. Starting from acoustic recordings of reiterant "ma" syllables, the perceptual importance of the following three factors was investigated: F0 height, F0 contour, and speech rhythm. For speakers with typically low or high voices F0 height turned out to be a highly relevant cue in speaker identification. For all speakers F0 contour was of secondary importance, whereas speech rhythm had a small but consistent influence on recognition rates. It could be inferred that remaining factors alone (mainly global spectral information) would yield recognition scores of approximately 50%. Consistent with previous investigations, the relevance of perceptual cues in the recognition of familiar voices was shown to be not hierarchically fixed, but to depend on speaker-specific voice characteristics.


Three-dimensional solubility parameters and their use in characterising the permeation of drugs through the skin.
The physico-chemical properties of drug substances are major determinants of their transdermal absorption. In the present study the concept of the three-dimensional solubility parameters of Hansen was applied in conjunction with the Bagley projection to describe the permeation of drugs and model substances through the skin. Drug permeation data from the literature were compared with the calculated solubility parameters of the drugs. It was demonstrated that the permeation of drugs can be estimated by their position in the Bagley diagram. There is a linear correlation between the logarithm of the skin permeation of drugs and the exchange cohesive energy for the steroids testosterone, progesterone, hydrocortisone acetate, corticosterone, cortisone, and dexamethasone. A linear correlation can be confirmed for the permeation of glyceryl trinitrate, digitoxin, oestradiol, scopolamine, atropine, diethylcarbamazine, fentanyl, and chlorpheniramine. In the case of morphine, codeine, sufentanil, meperidine and hydromorphone there is a linear relationship, too.


Novel partial reduction of the humanized anti-cocaine mAb h2E2 for selective cysteine labeling.
Monoclonal antibodies are utilized for treating many diseases and disorders, as well as for basic research and development. Covalent labeling of mAbs is important for various antibody applications and creating antibody drug conjugates. Labeling at reactive lysine residues using lysine selective reagents is useful, but is non-selective and can interfere with antigen binding and interactions of the Fc antibody region. In this work, using an anti-cocaine mAb (h2E2), we utilized triphenylphosphine-3,3',3″-trisulfonic acid (TPPTS), and demonstrated for the first time reduction of disulfides in an antibody by TPPTS. More importantly, this reduction was very reproducible, limited, and selective, and permitted selective labeling of the antibody with a cysteine reactive fluorescent reagent, resulting in labeling of a few specific cysteines. Similar results were obtained using TCEP-agarose reduction. We demonstrated that both of these selective partial reduction methods gave rise to approximately two labels per mAb, mostly by selective reduction of the heavy chain to light chain disulfide bond, as demonstrated by non-reducing SDS-PAGE protein band analysis. Thus, convenient, reproducible, and selective mAb disulfide reduction was achieved under mild conditions. These labeled, partially reduced mAbs were characterized by differential scanning fluorimetry (DSF), detecting the incorporated fluorescein instead of an exogenously added dye, and for antigen (cocaine) binding by isothermal titration calorimetry (ITC). Both the structure and antigen binding of the mAb was maintained. This novel selective reduction and labeling is generally relevant to modification of antibodies and to future development of conjugated mAbs for experimental and therapeutic purposes.


Motor coordination can be fully understood only by studying complex movements.
In this chapter, we use the sit-up to illustrate the complexity of coordination in movements that involve many muscles, joints, degrees of freedom, and high levels of muscle activity. Complex movements often involve the body axis. In addition to the intentional, focal part of any voluntary movement, complex movements also include "associated movements" that are not consciously controlled, but are necessary for the movement to succeed. Some associated movements serve a purpose, and others may not. During sitting up, the leg-lift is a purposive associated movement, whereas three-joint flexion is a non-purposive associated movement. The control of complex movements is also likely to be complex and, we argue, is hierarchically controlled. Associated movements may, themselves, be hierarchically organized and triggered by lower brain structures, local changes in neuronal excitability, and sensory feedback. Complex movements typically involve a high level of mobility. Because this mobility can lead to instability, anticipatory postural adjustments, a type of purposive associated movement, are commonly used to regulate posture. Thus, a number of important aspects of motor coordination can only be revealed by the study of complex movements.


Acetylation and phosphorylation of SRSF2 control cell fate decision in response to cisplatin.
SRSF2 is a serine/arginine-rich protein belonging to the family of SR proteins that are crucial regulators of constitutive and alternative pre-mRNA splicing. Although it is well known that phosphorylation inside RS domain controls activity of SR proteins, other post-translational modifications regulating SRSF2 functions have not been described to date. In this study, we provide the first evidence that the acetyltransferase Tip60 acetylates SRSF2 on its lysine 52 residue inside the RNA recognition motif, and promotes its proteasomal degradation. We also demonstrate that the deacetylase HDAC6 counters this acetylation and acts as a positive regulator of SRSF2 protein level. In addition, we show that Tip60 downregulates SRSF2 phosphorylation by inhibiting the nuclear translocation of both SRPK1 and SRPK2 kinases. Finally, we demonstrate that this acetylation/phosphorylation signalling network controls SRSF2 accumulation as well as caspase-8 pre-mRNA splicing in response to cisplatin and determines whether cells undergo apoptosis or G(2)/M cell cycle arrest. Taken together, these results unravel lysine acetylation as a crucial post-translational modification regulating SRSF2 protein level and activity in response to genotoxic stress.


Electrophysiologic actions and interactions between lysophosphatidylcholine and lidocaine in the nonsteady state: the match between multiphasic arrhythmogenic mechanisms and multiple drug effects in cardiac Purkinje fibers.
The effects and interactions between an arrhythmogenic intervention and an antiarrhythmic drug on active and passive cellular properties relevant to excitability were studied with multiple microelectrode methods and rapid online data analysis in cardiac Purkinje fibers. The arrhythmogenic intervention was superfusion with lysophosphatidylcholine (LPC), a metabolite that accumulates in the ischemic myocardium; and the antiarrhythmic drug was lidocaine. LPC (10-20 microM) initially increased excitability as manifested by a decreased threshold current and, when tested, by a downward shift in nonnormalized strength- and charge-duration curves. Normalized strength- and charge-duration curves suggested altered passive properties to be primarily responsible for increased excitability. Cable analysis showed LPC to increase significantly input resistance, membrane resistance, time constant and length constant; current-voltage relationships showed LPC to decrease chord and slope conductances over the subthreshold range. Lidocaine (4 micrograms/ml) decreased excitability both by depressing the sodium system and by directly countering the effects of LPC on conductance and related properties. LPC subsequently decreased excitability by depressing the sodium system and, in this phase, the further depressant effect of lidocaine on the sodium system predominated. Lidocaine could normalize action potentials that were prolonged or had two stable steady states after LPC, at times retarded LPC-induced inexcitability and could render the tissue inexcitable to intracellular point stimulation but not to extracellular stimulation. Interactions between the arrhythmogenic and pharmacologic interventions affected net excitability by altering the matrix of active and passive cellular properties in time. The results are relevant to the development of a rational matrical approach to understanding drug action and treating arrhythmias.


Demonstration in vitro of cell mediated immunity to Epstein-Barr virus in cotton-top tamarins.
In the course of developing an effective Epstein-Barr (EB) virus vaccine, the immune responses in cotton-top tamarins to a tumourigenic dose of EB virus were studied. Cell mediated responses were measured using a tissue culture 'growth inhibition' assay where peripheral blood lymphocytes were tested for their ability to inhibit the outgrowth of autologous EB virus transformed lymphoblastoid cells. This system has previously been recognized as a very sensitive assay for detecting cell-mediated responses to EB virus in man. Using this assay no cell-mediated immunity was detected up to the time of death in two tamarins following injection with a tumourigenic dose of EB virus. However, two other animals which had recovered from tumours induced by a first dose of EB virus 18 months previously when subsequently re-stimulated with a second tumourigenic dose did exhibit cell-mediated responses. These latter animals remained healthy following the re-challenge and did not show evidence of EB virus-induced disease.


[Reactions of humoral and cellular immunity in peptic ulcer in relation to the condition of the gastric mucosa].
A total of 160 persons including 50 patients with duodenal ulcer (DU) and 38 with gastric ulcer (GU) were examined for antibodies to the parietal cells of the stomach (PCA) and cell cellular immunity responses to autologous antigen from the gastric mucosa. It was shown that both PCA of the stomach and cell immunity responses in patients with GU and DU occurred in an insignificant number of cases. No differences were revealed in the humoral and cellular immunity in GU and DU persons with and without concomitant gastritis or in the stage of exacerbation and ulcer cicatrization.


Trypsin and rosmarinic acid reduce the toxicity of Micrurus fulvius venom in mice.
CONTEXT. Antivenom is expensive and not always available, so alternative treatments are being investigated. OBJECTIVE. The efficacy of trypsin or rosmarinic acid (RA) in treating Micrurus fulvius in a murine model is determined. MATERIALS AND METHODS. randomized controlled blinded study. Fifty mice (20-30 g). Study groups: Intraperitoneal injections of: 1) 2 mg/kg M. fulvius venom (approximately twice the LD50 for mice; n = 10); 2) 2 mg/kg M. fulvius venom incubated in vitro for 1 h prior to injection with RA at a 1:10 ratio (n = 17); 3) 2 mg/kg M. fulvius venom incubated in vitro for 1 h prior to injection with 1 mg of trypsin (n = 17); 4)1 mg trypsin IP without venom (n = 3); and 5) RA IP without venom (n = 3). time to toxicity (respiratory distress (< 25 breaths/min.), loss of spontaneous locomotor activity, or inability to upright self). Time to toxicity using Tukey-Kramer HSD; Survival to 4, 6, and 12 h using Chi-square analysis. RESULTS. Onset of toxicity: venom + saline, 120.3 + 64.4 min; venom + rosmarinic acid, 238.1 ± 139.2 min (p = 0.15 relative to venom + saline); venom + trypsin, 319.7 + 201.0 min (p = 0.007 relative to venom + saline). Venom + trypsin but not venom + RA survival to 4 h was significant compared to venom + saline (p = 0.023). Two mice in the venom + trypsin group and one mouse in the venom + RA group survived to 12 h. Mice receiving trypsin without venom or RA without venom survived to 12 h without toxicity. Discussion. This work suggests that trypsin and RA may have efficacy in treatment M. fulvius envenomation. CONCLUSION. In vitro neutralization of M. Fulvius venom by trypsin justifies progressing to an in vivo model in future studies.


ApoE4 and Connectivity-Mediated Spreading of Tau Pathology at Lower Amyloid Levels.
For the Alzheimer disease (AD) therapies to effectively attenuate clinical progression, it may be critical to intervene before the onset of amyloid-associated tau spreading, which drives neurodegeneration and cognitive decline. Time points at which amyloid-associated tau spreading accelerates may depend on individual risk factors, such as apolipoprotein E ε4 (ApoE4) carriership, which is linked to faster disease progression; however, the association of ApoE4 with amyloid-related tau spreading is unclear. To assess if ApoE4 carriers show accelerated amyloid-related tau spreading and propose amyloid positron emission tomography (PET) thresholds at which tau spreading accelerates in ApoE4 carriers vs noncarriers. This cohort study including combined ApoE genotyping, amyloid PET, and longitudinal tau PET from 2 independent samples: the Alzheimer's Disease Neuroimaging Initiative (ADNI; n = 237; collected from April 2015 to August 2022) and Avid-A05 (n = 130; collected from December 2013 to July 2017) with a mean (SD) tau PET follow-up time of 1.9 (0.96) years in ADNI and 1.4 (0.23) years in Avid-A05. ADNI is an observational multicenter Alzheimer disease neuroimaging initiative and Avid-A05 an observational clinical trial. Participants classified as cognitively normal (152 in ADNI and 77 in Avid-A05) or mildly cognitively impaired (107 in ADNI and 53 in Avid-A05) were selected based on ApoE genotyping, amyloid-PET, and longitudinal tau PET data availability. Participants with ApoE ε2/ε4 genotype or classified as having dementia were excluded. Resting-state functional magnetic resonance imaging connectivity templates were based on 42 healthy participants in ADNI. Mediation of amyloid PET on the association between ApoE4 status and subsequent tau PET increase through Braak stage regions and interaction between ApoE4 status and amyloid PET with annual tau PET increase through Braak stage regions and connectivity-based spreading stages (tau epicenter connectivity ranked regions). The mean (SD) age was 73.9 (7.35) years among the 237 ADNI participants and 70.2 (9.7) years among the 130 Avid-A05 participants. A total of 107 individuals in ADNI (45.1%) and 45 in Avid-A05 (34.6%) were ApoE4 carriers. Across both samples, we found that higher amyloid PET-mediated ApoE4-related tau PET increased globally (ADNI b, 0.15; 95% CI, 0.05-0.28; P = .001 and Avid-A05 b, 0.33; 95% CI, 0.14-0.54; P < .001) and in earlier Braak regions. Further, we found a significant association between ApoE4 status by amyloid PET interaction and annual tau PET increases consistently through early Braak- and connectivity-based stages where amyloid-related tau accumulation was accelerated in ApoE4carriers vs noncarriers at lower centiloid thresholds, corrected for age and sex. The findings in this study indicate that amyloid-related tau accumulation was accelerated in ApoE4 carriers at lower amyloid levels, suggesting that ApoE4 may facilitate earlier amyloid-driven tau spreading across connected brain regions. Possible therapeutic implications might be further investigated to determine when best to prevent tau spreading and thus cognitive decline depending on ApoE4 status.


Mutation analysis of autosomal dominant polycystic kidney disease genes in Han Chinese.
Autosomal dominant polycystic kidney disease (ADPKD) is caused by mutations in two genes, PKD1 and PKD2. The complexity of these genes, particularly PKD1, has complicated genetic screening, though recent advances have provided new opportunities for amplifying these genes. In the Han Chinese population, no complete mutational analysis has previously been conducted across the entire span of PKD1 and PKD2. Here, we used single-strand conformation polymorphism (SSCP) analysis to screen the entire coding sequence of PKD1 and PKD2 in 85 healthy controls and 72 Han Chinese from 24 ADPKD pedigrees. In addition to 11 normal variants, we identified 17 mutations (12 in PKD1 and 5 in PKD2), 15 of which were novel ones (11 for PKD1 and 4 for PKD2). We did not identify any seeming mutational hot spots in PKD1 and PKD2. Notably, we found several disease-associated C-T or G-A mutations that led to charge or hydrophobicity changes in the corresponding amino acids. This suggests that the mutations cause conformational alterations in the PKD1 and PKD2 protein products that may impact the normal protein functions. Our study is the first report of screenable mutations in the full-length PKD1 and PKD2 genes of the Han Chinese, and also offers a benchmark for comparisons between Caucasian and Han ADPKD pedigrees and patients.


Evidence for the bacterial origin of genes encoding fermentation enzymes of the amitochondriate protozoan parasite Entamoeba histolytica.
Entamoeba histolytica is an amitochondriate protozoan parasite with numerous bacterium-like fermentation enzymes including the pyruvate:ferredoxin oxidoreductase (POR), ferredoxin (FD), and alcohol dehydrogenase E (ADHE). The goal of this study was to determine whether the genes encoding these cytosolic E. histolytica fermentation enzymes might derive from a bacterium by horizontal transfer, as has previously been suggested for E. histolytica genes encoding heat shock protein 60, nicotinamide nucleotide transhydrogenase, and superoxide dismutase. In this study, the E. histolytica por gene and the adhE gene of a second amitochondriate protozoan parasite, Giardia lamblia, were sequenced, and their phylogenetic positions were estimated in relation to POR, ADHE, and FD cloned from eukaryotic and eubacterial organisms. The E. histolytica por gene encodes a 1,620-amino-acid peptide that contained conserved iron-sulfur- and thiamine pyrophosphate-binding sites. The predicted E. histolytica POR showed fewer positional identities to the POR of G. lamblia (34%) than to the POR of the enterobacterium Klebsiella pneumoniae (49%), the cyanobacterium Anabaena sp. (44%), and the protozoan Trichomonas vaginalis (46%), which targets its POR to anaerobic organelles called hydrogenosomes. Maximum-likelihood, neighbor-joining, and parsimony analyses also suggested as less likely E. histolytica POR sharing more recent common ancestry with G. lamblia POR than with POR of bacteria and the T. vaginalis hydrogenosome. The G. lamblia adhE encodes an 888-amino-acid fusion peptide with an aldehyde dehydrogenase at its amino half and an iron-dependent (class 3) ADH at its carboxy half. The predicted G. lamblia ADHE showed extensive positional identities to ADHE of Escherichia coli (49%), Clostridium acetobutylicum (44%), and E. histolytica (43%) and lesser identities to the class 3 ADH of eubacteria and yeast (19 to 36%). Phylogenetic analyses inferred a closer relationship of the E. histolytica ADHE to bacterial ADHE than to the G. lamblia ADHE. The 6-kDa FD of E. histolytica and G. lamblia were most similar to those of the archaebacterium Methanosarcina barkeri and the delta-purple bacterium Desulfovibrio desulfuricans, respectively, while the 12-kDa FD of the T. vaginalis hydrogenosome was most similar to the 12-kDa FD of gamma-purple bacterium Pseudomonas putida. E. histolytica genes (and probably G. lamblia genes) encoding fermentation enzymes therefore likely derive from bacteria by horizontal transfer, although it is not clear from which bacteria these amebic genes derive. These are the first nonorganellar fermentation enzymes of eukaryotes implicated to have derived from bacteria.


Lateral femoral cutaneous nerve impairment after direct anterior approach for total hip arthroplasty.
The anterior supine approach for total hip arthroplasty (THA) offers the advantage of operating through a true intravascular and intranervous plane, but it places the lateral femoral cutaneous nerve at risk. The purpose of this study was to identify the incidence of and impairment relating to injury of the lateral femoral cutaneous nerve. We performed a retrospective chart review of 81 hips undergoing anterior supine THA from November 2005 through May 2007 to determine operative time, estimated blood loss, fluoroscopic time, type of anesthesia used, intraoperative complications, and postoperative systemic and wound complications. Postoperative radiographs were evaluated for leg-length discrepancy, acetabular inclination and anteversion, and femoral stem position. Patients were reassessed at 6 weeks, 3 months, 6 months, 1 year, and 2 years. At each visit, patients were questioned about numbness or paresthesias in the distribution of the lateral femoral cutaneous nerve; if present, the patient outlined the area with a marking pen. This area was photographed, and data were collected. No hip had frank numbness; 12 hips (14.8%) had paresthesias. For those 12, symptoms resolved in 4 by 6 months, in 6 by 1 year, and in 10 (83.3%) by 2 years; 2 remained unresolved. No significant difference was found between patients with and without paresthesias or between patients with resolved or unresolved paresthesias. Impaired sensation did not appear to affect functional outcome or Harris Hip Score. Incision position, dissection plane, retractor placement, tension and soft tissue handling, and surgeon experience may affect incidence of injury to the lateral femoral cutaneous nerve.


Evaluation of a hematology analyzer with canine and feline blood.
A semiautomatic electronic blood cell counter (Sysmex F-800:Toa Medical Electronics Europa Gmbh, Hamburg, Germany) was evaluated using canine and feline blood, following the International Committee for Standardization in Hematology protocol (ICSH, 1984). Precision and overall reproducibility were acceptable for all the parameters studied except for the feline platelet count, in which overlapping of erythrocyte and platelet populations prohibited determination of an accurate platelet count. Since carry-over from canine hematocrit values and platelet counts and from feline hematocrit values was unsatisfactory, the use of a blank diluent sample between different analyses was necessary. Linearity of the analyzer was acceptable in the studied range. Thirty canine and feline blood samples were analyzed using the Sysmex F-800 and a manual method. Correlations between both methods were acceptable for all the parameters, except for feline platelet count and erythrocyte indices for both species. In the storage study, red blood cell count and hemoglobin concentration were the parameters with the longest stability (72 hours at 4 degrees C and 25 degrees C) in both species. A statistically significant increase in MCV was obtained at 12 hours post-extraction in canine samples stored at 25 degrees C and at 24 hours in refrigerated samples. Feline leucocyte counts showed a downward trend at 12 hours post-extraction at both temperatures. Canine platelet count decreased significantly at 6 hours post-extraction in samples stored at 4 degrees C. During the evaluation period, Sysmex F-800 was user friendly and appeared well suited for routine canine and feline blood cell analysis.


Trans-catheter aortic valve implantation for non-classical indications.
Trans-catheter aortic valve implantation (TAVI) has emerged as a novel therapeutic approach for patients with severe tricuspid aortic stenosis (AS) not suitable for aortic valve replacement. To describe our initial single-center experience with TAVI in patients with "off-label" indications. Between August 2008 and December 2011 we performed TAVI in 186 patients using trans-femoral, transaxillary, trans-apical and trans-aortic approaches. In 11 patients (5.9%) TAVL was undertaken due to: a) pure severe aortic regurgitation (AR) (n = 2), b) prosthetic aortic valve (AV) failure (n = 5), c) bicuspid AV stenosis (n = 2), and d) prosthetic valve severe mitral regurgitation (MR) (n = 2). Implantation was successful in all: six patients received a CoreValve and five patients an Edwards-Sapien valve. In-hospital mortality was 0%. Valve hemodynamics and function were excellent in all patients except for one who received an Edwards-Sapien that was inside a Mitroflow prosthetic AV and led to consistently high trans-aortic gradients. No significant residual regurgitation in AR and MR cases was observed. TAVI is a good alternative to surgical AV replacement in high risk or inoperable patients with severe AS. TAVI for non-classical indications such as pure AR, bicuspid AV, and failed prosthetic aortic and mitral valves is feasible and safe and may be considered in selected patients.


Percutaneous ultrasonic versus surgical removal of kidney stones.
Kidney stones may be removed without using a surgical incision by a combination of techniques and skills recently developed in the fields of urology and radiology. Percutaneous access to the kidney is established under fluoroscopic control. A guide wire placed into the renal pelvis allows a nephroscope to be inserted and the collecting system visualized. A long hollow metal probe is advanced through the nephroscope and placed in contact with the stone. This probe conducts the ultrasonic energy. The stone absorbs the energy and breaks into fine granules which are evacuated by suction. Percutaneous lithotomy was performed upon 300 of 302 consecutive patients bearing stones which required removal. Targeted calculi were successfully removed from 97 per cent of these patients. Eighty-one per cent had complete stone removal at the primary procedure; 57 patients required additional percutaneous manipulations under local or general anesthesia. The previous 100 patients with kidney stones underwent surgical removal of the calculi. Complications associated with percutaneous stone removal included postoperative fever, delayed urinary bleeding, ureteral edema, symptomatic urinary extravasation and water intoxication. Only 16 patients with complications required prolonged hospitalization or readmission. No complications required surgical management; no deaths occurred. Anesthetic times were similar for both groups, averaging 152 +/- 4 (S. E. M.) minutes for PL and 193 +/- 8 minutes for SL. Hospital recovery days averaged 5.7 +/- 0.3 for PL and 8.4 +/- 0.5 for SL (p less than 0.01). Associated costs averaged $7,203 +/- 255 for PL and $8,849 +/- 660 for SL (p less than 0.01). The number of narcotic administrations per patient (days one to five postoperatively) averaged 9.9 +/- 0.7 for the PL group and 16.8 +/- 0.8 for the SL group (p less than 0.01). The average patient in the PL group felt capable of full activity 2.0 +/- 0.2 weeks after stone removal, whereas no patient who experienced previous SL recalls a recovery period of less than three weeks (p less than 0.01). Most upper urinary tract calculi may be removed cost-effectively using a percutaneous approach. Patients may expect a rapid convalescence with diminished pain.