[Alcohol-induced toxic hepatitis--a "free radical" associated disease. Lowering fatality by adjuvant antioxidant therapy].
Toxic liver diseases coincide with oxidative stress correlating positively with the seriousness of the course of disease. For the purpose of elucidating the pathogenic significance of an increased radical generation. 56 patients suffering from acute alcohol-toxic hepatitis of the clinical grade of seriousness B and C according to Child/Pugh were classified randomly into antioxidant subgroups (n = 31) and control groups (= 25). The basis therapy being identical, the patients of the antioxidant group received additionally 600 mg of D-alpha tocopherol per day, 200 micrograms of selenium and 12 mg of zinc. Due to the supplementation of antioxidants there were quicker significant changes in the concentration of bilirubin, malondialdehyde and of ammonia in the serum. In comparison with the control group the length of stay in hospital could be reduced by 6 days. In the control group the mortality rates amounted to 40% (10 of 25), in the antioxidant group to 6.5% (2 of 31). The results confirm the pathogenic significance of oxidative stress in alcohol-toxic liver disease because a distinct improvement of prognosis could be achieved by using a low-cost adjuvant antioxidant supplementation.


Posttranslational protein modification by amino acid addition in intact and regenerating axons of the rat sciatic nerve.
Experiments were performed to determine whether posttranslational addition of amino acids to axonal proteins occurs in axons of the rat sciatic nerve. Two ligatures were placed 1 cm apart on sciatic nerves. Six days later, segments proximal to each ligature were removed, homogenized, centrifuged at 150,000 X g, and analyzed for the ability to incorporate 3H-amino acids into proteins. No incorporation of amino acids into proteins was found in the high-speed supernatant, but when the supernatant was passed through a Sephacryl S-200 chromatography column (removing molecules less than 20 kD), [3H]arginine, lysine, leucine and aspartic acid were incorporated into proteins in both proximal and distal nerve segments. Small but consistently greater amounts of radioactivity were incorporated into proteins in proximal segments compared with distal segments, indicating that the components necessary for the reaction are transported axonally. This reaction represents the posttranslational incorporation of a variety of amino acids into proteins of rat sciatic nerve axons. Other experiments showed that the incorporation of amino acids into proteins is by covalent bonding, that the amino acid donor is likely to be tRNA, and that the reaction is inhibited in vivo by a substance whose molecular mass is less than 20 kD. This inhibition is not affected by incubation with physiological concentrations of unlabeled amino acids, by boiling, or by treatment with Proteinase K. When the axonally transported component of the reaction was determined in regenerating nerves, the amount of incorporation of amino acids into protein was 15-150 times that in intact nerves.(ABSTRACT TRUNCATED AT 250 WORDS)


Infection fatality rate of SARS-CoV2 in a super-spreading event in Germany.
A SARS-CoV2 super-spreading event occurred during carnival in a small town in Germany. Due to the rapidly imposed lockdown and its relatively closed community, this town was seen as an ideal model to investigate the infection fatality rate (IFR). Here, a 7-day seroepidemiological observational study was performed to collect information and biomaterials from a random, household-based study population. The number of infections was determined by IgG analyses and PCR testing. We found that of the 919 individuals with evaluable infection status, 15.5% (95% CI:[12.3%; 19.0%]) were infected. This is a fivefold higher rate than the reported cases for this community (3.1%). 22.2% of all infected individuals were asymptomatic. The estimated IFR was 0.36% (95% CI:[0.29%; 0.45%]) for the community and 0.35% [0.28%; 0.45%] when age-standardized to the population of the community. Participation in carnival increased both infection rate (21.3% versus 9.5%, p < 0.001) and number of symptoms (estimated relative mean increase 1.6, p = 0.007). While the infection rate here is not representative for Germany, the IFR is useful to estimate the consequences of the pandemic in places with similar healthcare systems and population characteristics. Whether the super-spreading event not only increases the infection rate but also affects the IFR requires further investigation.


Glucose starvation suppresses gastric cancer through targeting miR-216a-5p/Farnesyl-Diphosphate Farnesyltransferase 1 axis.
Fasting mimic diet is an effect approach for gastric cancer (GC) treatment. Exploring mechanisms of glucose deprivation-mediated GC suppression is required to develop novel therapeutic regimens. Farnesyltransferase 1 (FDFT1), as a novel target in basic research, has been reported to regulate malignant progression in some types of cancer. However, biological functions of FDFT1 in GC are still unclear. This study focused on biological functions of FDFT1 in GC and the association between glucose starvation (GS) and FDFT1. The data derived from the Kaplan-Meier Plotter database were collected to identify the relationship between survival time and FDFT1 expression levels of GC patients. Bioinformatic analysis was performed to explore the biological functions of FDFT1. The expression levels of targeted genes and microRNAs (miRNAs) were detected with immunohistochemistry, quantitative real-time PCR and western blot. Malignant behaviors were measured using cell counting, cell counting kit-8, 5-ethynyl-2-deoxyuridine, wound healing, invasion transwell assays in vitro and constructions of subcutaneous and lung-metastatic tumors in vivo. The glycolysis of GC cells was determined by a series of metabolites, including lactate acid, pyruvic acid, ATP production, rates of glucose uptake, extracellular acidification rate and oxygen consumption rate. FDFT1 was downregulated in GC and negatively correlated with pathological T stage, pathological TNM stage and cancer differentiation. High expression of FDFT1 also indicated better prognosis of GC patients. FDFT1 upregulation attenuated proliferation, migration and invasion of GC. miR-216a-5p was identified as a critical suppressor of FDFT1 expression and miR-216a-5p/FDFT1 axis regulated malignant behaviors and glycolysis of GC cells. GS suppressed malignant behaviors of GC by targeting miR-216a-5p/FDFT1 axis both in vitro and in vivo. This study illustrated novel mechanisms by which GS effectively suppresses GC. FDFT1 may become a potential prognostic indicator and novel target of GC therapy.


Platelet alterations in porcine stress syndrome.
Platelets in blood collected from pigs identified as normal (21) or stress susceptible (25) on the basis of their response to halothane challenge, were subjected to electron microscopic examination in order to test whether the ultrastructural features of stress susceptible pigs exhibit any deviation from those of normal. The most striking feature of the platelets from stress susceptible pigs was the extent of dilatation of the open canalicular system (OCS). The difference in platelet morphology between normal and stress susceptible pigs was consistent regardless of the anticoagulants used for blood collection indicating that the platelet alteration may be an inherent component of the porcine stress syndrome.


Practice to evidence to practice: closing the loop with IT.
Greater use of existing and developing IT resources can close the loop, creating an effective cycle of practice to evidence and back to practice.


Impact of contrast medium osmolality on the risk of acute kidney injury after transcatheter aortic valve implantation: insights from the Magna Graecia TAVI registry.
Acute kidney injury (AKI) after transcatheter aortic valve implantation (TAVI) is frequent and associated with adverse outcomes and mortality; to date, in such setting of patients there is no consistent evidence that either low-osmolar contrast media (LOCM) or iso-osmolar contrast medium (IOCM) are superior to the other in terms of renal safety. 697 consecutive patients not in hemodialysis treatment who underwent TAVI (327 males, mean age 81.01 ± 5.75 years, mean european system for cardiac operative risk evaluation II 6.17 ± 0.23%) were enrolled. According to osmolality of the different iodinated CM, the population was divided in 2 groups: IOCM (n = 370) and LOCM group (n = 327). Preoperatively, 40.54% of patients in IOCM vs 39.14% in LOCM group (p = 0.765) suffered from chronic kidney disease (CKD). The incidence of AKI was significantly lower with IOCM (9.73%) than with LOCM (15.90%; p = 0.02), and such significant difference (p < 0.001) in postprocedural change of renal function parameters persisted at discharge too. The incidence of AKI was also significantly lower with IOCM in younger patients, without diabetes, anemia, coronary artery disease history, CKD, chronic or persistent atrial fibrillation, left ventricular ejection fraction ≤35%, and in patients with low operative mortality risk scores, receiving lower amounts of dye (p < 0.05 for all). Importantly, multivariate analysis identified LOCM administration as an independent risk factor for both AKI (p = 0.006) and 1-year mortality (p = 0.001). The use of IOCM have a favorable impact on renal function with respect to LOCM, but it should be considered especially for TAVI patients at lower AKI risk.


Signal formation in the halobacterial photophobic response mediated by a fourth retinal protein (P480).
Halobacterial cells swim forward by clockwise, and backward by counterclockwise, rotation of their flagella. The changes of direction of rotation occur statistically and can be quantitatively described by a four-state model of the motor. Stimulation of the cells with blue light induces the formation of a signal that causes the motor to switch the direction of rotation. The results of step-up and flash experiments led to a kinetic equation that describes the signal formation as a photocatalytic process. The stimulating blue light is sensed either by sensory rhodopsin in the presence of green background light or by protein P480, which has a maximum in the action spectrum around 480 nm. P480, but not sensory rhodopsin, is synthesized by the cells constitutively, and both pigments together allow the cells to find optimal conditions during aerobic and phototrophic growth. The work presented here was reported at the U.S.-Israel Binational Science Foundation Meeting in Jerusalem, March 1986.


Alpha-neo-endorphin-like immunoreactivity in the cat brain stem.
This paper examines the distribution of fibers and cell bodies containing alpha-neo-endorphin in the cat brain stem by using an indirect immunoperoxidase technique. A high or moderate density of immunoreactive cell bodies was found in the superior central nucleus, nucleus incertus, dorsal tegmental nucleus, nucleus of the trapezoid body, and in the laminar spinal trigeminal nucleus, whereas a low density of such perikarya was observed in the inferior colliculus, nucleus praepositus hypoglossi, dorsal nucleus of the raphe, nucleus of the brachium of the inferior colliculus, and in the nucleus of the solitary tract. The highest density of immunoreactive fibers was found in the substantia nigra, dorsal motor nucleus of the vagus, nucleus coeruleus, lateral tegmental field, marginal nucleus of the brachium conjunctivum, and in the inferior and medial vestibular nuclei. These results indicate that alpha-neo-endorphin is widely distributed in the cat brain stem and suggest that the peptide could play an important role in several physiological functions, e.g., those involved in respiratory, cardiovascular, auditory, and motor mechanisms.


Atypical osteoid osteomas.
Osteoid osteoma is a very painful benign bone tumor that affects young adults. It is exceptional before 5 and after 30 years of age. It can be seen in any part of the skeleton. Presentation of the tumor may be intracortical, juxta-cortical or cancellous. In long bones, the tumor is frequently intracortical, and in the spine, it is usually located at the posterior elements. Osteoid osteomas having radiologic and clinical features other than classical presentation of osteoid osteoma are called atypical osteoid osteomas. Atypical osteoid osteomas are important because the diagnosis and treatment are often complicated in these cases. Misdiagnosis with prolonged impairment and sometimes overtreatment appears as a major problem concerning atypical osteoid osteomas. This paper gives brief general information about the classical presentation of osteoid osteoma, discusses the pathogenesis of the lesion and focuses on the clinical presentation, radiologic features and characteristics of atypical osteoid osteomas as well as their treatment modalities.


The adaptive immune system as a fundamental regulator of adipose tissue inflammation and insulin resistance.
Over the past decade, chronic inflammation in visceral adipose tissue (VAT) has gained acceptance as a lead promoter of insulin resistance in obesity. A great deal of evidence has pointed to the role of adipokines and innate immune cells, in particular, adipose tissue macrophages, in the regulation of fat inflammation and glucose homeostasis. However, more recently, cells of the adaptive immune system, specifically B and T lymphocytes, have emerged as unexpected promoters and controllers of insulin resistance. These adaptive immune cells infiltrate obesity expanded VAT and through cytokine secretion and macrophage modulation dictate the extent of the local inflammatory response, thereby directly impacting insulin resistance. The remarkable ability of our adaptive immune system to regulate insulin sensitivity and metabolism has unmasked a novel physiological function of this system, and promises new diagnostic and therapeutic strategies to manage the disease. This review highlights critical roles of adipose tissue lymphocytes in governing glucose homeostasis.


Fibrocartilage extracellular matrix augmented demineralized bone matrix graft repairs tendon-to-bone interface in a rabbit tendon reconstruction model.
Current tendon/ligament reconstructions integrate via scar tissue rather than proper bone-tendon interface regeneration, which affects graft longevity, changes in bone tunnel size, and functional outcomes. The purpose of this study was to develop a functional demineralized bone matrix (DBM) + fibrocartilage extracellular matrix (FCECM) composite scaffold, characterize its physicochemical properties, and evaluate its efficacy in repairing tendon-bone interface in a rabbit tendon reconstruction model. Solubilized FCECM was loaded and crosslinked on to DBM scaffolds via gamma-irradiation to create DBM + FCECM scaffolds. The resulting scaffold showed interconnected pores coated with FCECM and protein cargo similar to FCECM. The addition of FCECM modified the physicochemical properties of the DBM scaffold, including microstructure, biochemical composition, mechanical strength, thermodynamic properties, and degradation period. The DBM + FCECM scaffold was biocompatible for mesenchymal stem cells (MSCs) and resulted in elevation of fibrochondrogenic gene markers compared to DBM scaffolds in vitro. In vivo implantation of DBM + FCECM scaffold resulted in neofibrocartilage formation, better pullout strength, and less bone tunnel widening compared to DBM only group in a rabbit tendon reconstruction model. In conclusion, the FCECM augmented DBM scaffold repairs the tendon-bone interface with osseous-fibrocartilage tissue, which may be utilized to improve current tendon reconstruction surgeries.


A musculoskeletal modelling approach to explain sit-to-stand difficulties in older people due to changes in muscle recruitment and movement strategies.
By 2050 the proportion of over 65s is predicted to be 20% of the population. The consequences of an age-related reduction in muscle mass have not been fully investigated and, therefore, the aim of the present study was to quantify the muscle and joint contact forces using musculoskeletal modelling, during a sit-to-stand activity, to better explain difficulties in performing everyday activities for older people. A sit-to-stand activity with and without the use of arm rests was observed in ninety-five male participants, placed into groups of young (aged 18-35 years), middle-aged (aged 40-60 years) or older adults (aged 65 years and over). Older participants demonstrated significantly lower knee extensor and joint forces than the young when not using arm rests, compensating through elevated hip extensor and ankle plantarflexor muscle activity. The older group were also found to have higher shoulder joint contact forces whilst using arm rests. This tendency to reorganise muscle recruitment to include neighbouring groups or other parts of the body could make everyday activities more susceptible to age-related functional decline. Reductions in leg strength, via age- or atrophy- related means, creates increased reliance on the upper body and may result in further lower limb atrophy through disuse. The eventual decline of upper body function reduces strength reserves, leading to increased vulnerability, dependence on others and risk of institutionalisation.


Ramsay Hunt syndrome in dentatorubral-pallidoluysian atrophy.
We report a case of Ramsay Hunt syndrome which was clinically characterized by myoclonus epilepsy, cerebellar ataxia, convulsions, and dementia. Major necropsy findings were dentatorubral-pallidoluysian atrophy. Dentatorubral-pallidoluysian atrophy may be associated with a variety of clinical symptoms, amongst which Ramsay Hunt syndrome can be included.


Determination of lanthanides by source excited energy dispersive X-ray fluorescence (EDXRF) method after preconcentration with ammonium pyrrolidine dithiocarbamate (APDC).
A new analytical procedure for determination of lanthanides in environmental samples after chemical separation from major matrix elements on DOWEX 50W-X8 resin followed by preconcentration with chelating agent ammonium pyrrolidine dithiocarbamate (APDC) and analyses of thin targets by energy dispersive X-ray fluorescence (EDXRF) method using 109Cd as the source of excitation was presented. Characteristic Lalpha X-ray lines of the lanthanides were used for calculations of the net peak area and mass concentrations. The influence of pH value of the solution and addition of organic matter on the complexation was investigated. Percentage of recovery of each lanthanide after separation on DOWEX 50W-X8 resin was also determined. Accuracy of the method was tested on standard reference materials and real environmental samples (red mud material). For that purpose samples of standard reference materials and red mud were prepared as thick targets and directly analyzed (without the separation step) by EDXRF method using 241Am as the excitation source. In that case lanthanides concentrations were determined over their characteristic Kalpha X-ray lines and results were compared with those obtained after separation/preconcentration step described above. Results showed that selected lanthanides made stable complexes with APDC in the alkaline medium with the maximum recovery at pH=8. The presence of organic matter slightly modified the complexation by means of somewhat higher recovery percentage at pH lower than 7 and approx. 20% lower recovery at pH higher than 7. Recovery of the elements after separation on DOWEX 50W-X8 resin and preconcentration with APDC at pH=8 varied from 91.4% (Pr) to only 24.9% in the case of Dy. Concentrations of lanthanides measured in standard reference material and environmental samples of red mud after microwave digestion, separation on DOWEX 50W-X8 resin, preconcentration with APDC at pH=8 and recalculation on the percentage of recovery were in good agreement with certified values in the case of SRM as well as with the concentrations obtained by direct determination over Kalpha lines using 241Am excitation source in the case of red mud leading to the conclusion that presented method was applicable for the determination of lanthanides in real environmental samples.


Pretreatment of biomass by torrefaction and carbonization for coal blend used in pulverized coal injection.
To evaluate the utility potential of pretreated biomass in blast furnaces, the fuel properties, including fuel ratio, ignition temperature, and burnout, of bamboo, oil palm, rice husk, sugarcane bagasse, and Madagascar almond undergoing torrefaction and carbonization in a rotary furnace are analyzed and compared to those of a high-volatile coal and a low-volatile one used in pulverized coal injection (PCI). The energy densities of bamboo and Madagascar almond are improved drastically from carbonization, whereas the increase in the calorific value of rice husk from the pretreatment is not obvious. Intensifying pretreatment extent significantly increases the fuel ratio and ignition temperature of biomass, but decreases burnout. The fuel properties of pretreated biomass materials are superior to those of the low-volatile coal. For biomass torrefied at 300°C or carbonized at temperatures below 500°C, the pretreated biomass can be blended with coals for PCI.


[Construction of a mapping panel of human-rodent hybrid cells].
A clone panel of 27 human-Chinese hamster and 4 human-mouse somatic cell hybrids which contained as minimum five discriminating clones for any chromosome pairs was set up. Segregation analysis of 45 human chromosome-specific isoenzymes and PCR markers in hybrid clones allowed to demonstrate a possibility to apply the obtained panel for chromosome mapping of human genes.


The antibiotic thermorubin inhibits protein synthesis by binding to inter-subunit bridge B2a of the ribosome.
Thermorubin is a small-molecule inhibitor of bacterial protein synthesis, but relatively little is known about the molecular mechanism by which it blocks translation. The structure of the complex between thermorubin and the 70S ribosome from Thermus thermophilus reported here shows that thermorubin interacts with the ribosome in a way that is distinct from any other known class of ribosome inhibitor. Though it is structurally similar to tetracycline, it binds to the ribosome at an entirely different location-the interface between the small and large subunits that is formed by inter-subunit bridge B2a. This region of the ribosome is known to play a role in the initiation of translation, and thus, the binding site we observe is consistent with evidence suggesting that thermorubin inhibits the initiation stage of protein synthesis. The binding of thermorubin induces a rearrangement of two bases on helix 69 of the 23S rRNA, and presumably, this rearrangement blocks the binding of an A-site tRNA, thereby inhibiting peptide bond formation. Due in part to its low solubility in aqueous media, thermorubin has not been used clinically, although it is a potent antibacterial agent with low toxicity (Therapeutic Index>200). The interactions between thermorubin and the ribosome, as well as its adjacency to the observed binding sites of three other antibiotic classes, may enable the design of novel derivatives that share thermorubin's mode of action but possess improved pharmacodynamic properties.


Association between diabetes mellitus and hypertension with anthropometric indicators in older adults: results of the Mexican Health Survey, 2000.
To determine the association between anthropometric indicators of adiposity with type 2 diabetes mellitus (T2DM) and hypertension (HTN) in older adults. Cross-sectional study of participants of the Mexican Health Survey 2000 (MHS). Mexico, subjects recruited from the general community. The analytic sample included 7,322 adults who were > or = 60 years of age at the time of the survey. T2DM data were available on 6,994 individuals, who represent 95.5% of the original sample; data on HTN was available on 6,268 subjects, which accounted for 86.5% of the original sample. Type 2 diabetes mellitus and hypertension, as well as anthropometric indicators including body mass index (BMI), waist circumference (WC), and conicity index (CI). The prevalence of T2DM and HTN in this age group was 34.3% and 73.9%, respectively. After adjusting for other variables, the association between high WC and T2DM (OR = 1.59 95%CI = 1.26-2.01, P < 0.001) was stronger than the association with overweight (OR = 1.26, 95%CI = 1.01-1.58, P = 0.04) and obesity (OR = 1.38, 95%CI = 1.08-1.79, P < 0.01) using BMI, and slightly higher than tertile 2 of the CI (OR = 1.49, 95%CI = 1.20-1.88, P < 0.01), while tertile 3 showed a stronger association with T2DM (OR = 1.60, 95%CI = 1.22-2.08, P < 0.001). However, the association between obesity and HTN measured by BMI (OR = 1.98, 95%CI = 1.48-2.65, P < 0.001) was stronger than what was observed with overweight (OR = 1.42, 95%CI 1.13-1.77, P < 0.01), with high WC (OR = 1.62, 95%CI = 1.25-2.10, P < 0.001) and tertiles 2 and 3 of the CI (OR = 1.23, 95%CI = 0.99-1.55, P = 0.09); (OR = 1.53, 95%CI = 1.16-2.03, P < 0.01) respectively. BMI and abdominal obesity are significantly and independently associated with an increase in the prevalence of T2DM and HTN among older Mexican adults.


Activation of mammalian cyclic AMP phosphodiesterases by trypsin.
BHK fibroblasts contain two forms of cyclic AMP phosphodiesterase 3':5'-cyclic nucleotide 5'-nucleotidohydrolase EC 22.214.171.124) as analyzed by linear sucrose gradient fractionation; a 3.6-S form (peak I) and a 6.7-S form (peak II). Peak I is specific for cyclic AMP as substrate and displays Michaelis-Menten kinetics with an apparent Km of 2--3 micrometer. Peak II hydrolyzes cyclic GMP and displays anomalous kinetics for cyclic AMP hydrolysis. The activity of isolated peak II for cyclic AMP is increased by storage at 4 degrees C, treatment with trypsin, or treatment with rat brain and BHK fibroblast activator proteins. The activity of isolated peak I is unaffected by these conditions. Linear sucrose gradient fractionation demonstrates that activation of peak II by trypsin leads to the formation of a 3.6-S cyclic AMP-specific enzyme form, possibly peak I. In contrast to BHK fibroblasts (and most other mammalian tissues), rat uterus contains only one form of cyclic nucleotide phosphodiesterase on linear sucrose gradients, a 7-S form capable of hydrolyzing both cyclic AMP and cyclic GMP. Treatment of rat uterine supernatant with trypsin leads to the appearance of a 4-S, cyclic AMP-specific form with properties similar to that of BHK peak I. These data suggest that the kinetically complex, higher molecular weight cyclic nucleotide phosphodiesterases may consist of more than one catalytically active site and that multiple forms of the enzyme arise through dissociative mechanisms, possibly as a means of in vivo regulation.


The AGT and the GNB3 polymorphisms and insulin resistance in prehypertension.
This study surveyed the frequencies of single nucleotide polymorphisms (SNPs) M235T AGT and C825T GNB3, and their association with insulin resistance, other biochemical markers and qualitative variables in subjects with high normal blood pressure and/or prehypertension in the Greek population. 330 men and women of Greek origin were divided into 3 groups: a) hypertensive, b) prehypertensive and c) control group. These groups were genetically tested for these polymorphisms and insulin resistance with the HOMA index. No statistically significant differences were found among the polymorphisms of the compared groups. However, the ? allele carriers (CT/TT vs. CC) of the C825T polymorphism were associated with an increased BMI in all 3 groups (p=0.004). The HOMA index was higher in the hypertensive (p=0.006) and prehypertensive (p=0.016) versus the control group, and similar results were found for insulin (hypertensive vs. control p=0.012, prehypertensive vs. control p=0.001) without statistical significance between the first 2 groups (p=0.522). Additionally, there was a statistically significant difference between the control group and the hypertensive and prehypertensive groups regarding cholesterol (control vs. hypertensive p=0.001, control vs. prehypertension p=0.018) and triglycerides (control vs. hypertensive p=0.0001, control vs. prehypertension p=0.007). Differences were also noted between the control and the hypertensive group regarding the value of HDL (p=0.005) and LDL (p=0.013). This study failed to demonstrate a correlation between specific SNPs, blood pressure and insulin resistance in the 3 groups. However, T allele carriers of the polymorphism C825T were found to have an increased BMI. Similarly, increased insulin resistance and lipidemia were more common in the hypertensive and prehypertensive populations.


Upregulation of HIV-1 replication in chronically infected cells by ingenol derivatives.
We have previously reported that ingenol derivatives are highly potent inhibitors of human immunodeficiency virus type 1 (HIV-1) replication in acutely infected cells. In this study, however, we have found that some ingenol derivatives strongly enhance the replication of HIV-1 in chronically infected cells at nanomolar concentrations. One of the derivatives could activate nuclear factor kappa B (NF-kappa B), a potent inducer of HIV-1 replication, through the activation of protein kinase C (PKC). Whereas another derivative, which affected neither PKC nor NF-kappa B, significantly enhanced HIV-1 replication, suggesting that a PKC-independent mechanism may also exist in ingenol derivative-induced HIV-1 upregulation.


Early-Life Predictors of Fetal Alcohol Spectrum Disorders.
Fetal alcohol spectrum disorders (FASD) comprise the continuum of disabilities associated with prenatal alcohol exposure. Although infancy remains the most effective time for initiation of intervention services, current diagnostic schemes demonstrate the greatest confidence, accuracy, and reliability in school-aged children. Our aims for the current study were to identify growth, dysmorphology, and neurodevelopmental features in infants that were most predictive of FASD at age 5, thereby improving the timeliness of diagnoses. A cohort of pregnant South African women attending primary health care clinics or giving birth in provincial hospitals was enrolled in the project. Children were followed longitudinally from birth to 60 months to determine their physical and developmental trajectories (N = 155). Standardized protocols were used to assess growth, dysmorphology, and development at 6 weeks and at 9, 18, 42, and 60 months. A structured maternal interview, including estimation of prenatal alcohol intake, was administered at 42 or 60 months. Growth restriction and total dysmorphology scores differentiated among children with and without FASD as early as 9 months (area under the receiver operating characteristic curve = 0.777; P < .001; 95% confidence interval: 0.705-0.849), although children who were severely affected could be identified earlier. Assessment of developmental milestones revealed significant developmental differences emerging among children with and without FASD between 18 and 42 months. Mothers of children with FASD were significantly smaller, with lower BMIs and higher alcohol intake during pregnancy, than mothers of children without FASD. Assessment of a combination of growth, dysmorphology, and neurobehavioral characteristics allows for accurate identification of most children with FASD as early as 9 to 18 months.


Developing a biostatistical support system in a resource-restricted academic institution in Africa: making it happen.
In order to address and support biostatistics for health research, the Health Sciences Research Office of the University of the Witwatersrand sought to introduce training in biomedical statistics to sustain research and postgraduate education. The experiences encountered in setting up such statistical support in a limited resource, developing country are discussed here. Two cross-sectional surveys (a) statistical needs assessment (2009) and (b) feedback (2010-11) on the statistical support through biostatistics courses and consultations were conducted. These surveys were supplemented with information such as graduations, research publication output and costs of setting up the support. Seventy-three percent of respondents favoured short courses with "hands-on" practice. Eighty-nine percent agreed that these courses should be run and coordinated by the Health Sciences Research Office instead of the departments. There was use of varied statistical packages requiring one package for standardised support. The numbers of postgraduate students attending short courses in statistics increased from 2010 to 2012 as did the numbers attending statistical consultations. Graduations and publication outputs increased over this period of time although this may not be directly linked solely to the biostatistical support system introduced. There is a distinct need for biostatistics training in developing countries and the process described in this study could be replicated in any health sciences institution, especially in a resource-restricted environment.


Distribution of glutathione and glutathione-related enzyme systems in mitochondria and cytosol of cultured cerebellar astrocytes and granule cells.
The cellular and regional distribution of glutathione (GSH) and GSH-related enzyme systems involved in cellular defense against reactive oxygen species and electrophilic xenobiotics in the nervous system has been extensively studied. However, little is known about the subcellular distribution of GSH systems in brain tissue and cultured neural cells. The present study investigates the distribution of mitochondrial and cytosolic GSH and GSH-related enzymes in cultured cerebellar astrocytes and granule cells, and compares them with levels in the adult rat cerebellum. Cytosolic GSH levels and cytosolic activities of glutathione reductase (GR), glutathione peroxidase (GPX) and glutathione-S-transferase (GST) in astrocytes were 57, 153, 245, and 92% higher than those found in granule cells, respectively. In contrast, granule cells contained significantly higher mitochondrial GSH levels than astrocytes. Granule cells also demonstrated comparable mitochondria/cytosolic concentrations of GSH and GR, GPX and GST activities to those observed in the cerebellar tissue, whereas ratios in astrocytes were markedly lower. Although in vitro treatments with 100 microM ethacrynic acid depleted both cytosolic and mitochondrial GSH in cultured astrocytes and granule cells in a time-dependent fashion, cellular GSH in granule cells was more resistant to the GSH-depleting agent than astrocytes. These results suggest that although GSH and GSH-related enzymes are abundant in cytosolic compartments of astrocytes, mitochondrial pools are relatively small. Since brain mitochondria are sites of significant hydrogen peroxide generation, the mitochondrial localization of GSH and its associated enzymes in neural cells provide important defenses against toxic oxygen species in the nervous system. Differences in subcellular distribution of GSH systems in individual neural cell types may provide a basis for selective cellular and/or subcellular expression of neurotoxicity.


Nitric oxide interacts with mitochondrial complex III producing antimycin-like effects.
The effect of NO between cytochromes b and c of the mitochondrial respiratory chain were studied using submitochondrial particles (SMP) from bovine heart and GSNO and SPER-NO as NO sources. Succinate-cytochrome c reductase (complex II-III) activity (222 ± 4 nmol/min. mg protein) was inhibited by 51% in the presence of 500 μM GSNO and by 48% in the presence of 30 μM SPER-NO, in both cases at ~1.25 μM NO. Neither GSNO nor SPER-NO were able to inhibit succinate-Q reductase activity (complex II; 220 ± 9 nmol/min. mg protein), showing that NO affects complex III. Complex II-III activity was decreased (36%) when SMP were incubated with l-arginine and mtNOS cofactors, indicating that this effect is also produced by endogenous NO. GSNO (500 μM) reduced cytochrome b562 by 71%, in an [O2] independent manner. Hyperbolic increases in O2(•-) (up to 1.3 ± 0.1 nmol/min. mg protein) and H2O2 (up to 0.64 ± 0.05 nmol/min. mg protein) productions were observed with a maximal effect at 500 μM GSNO. The O2(•-)/H2O2 ratio was 1.98 in accordance with the stoichiometry of the O2(•-) disproportionation. Moreover, H2O2 production was increased by 72-74% when heart coupled mitochondria were exposed to 500 μM GSNO or 30 μM SPER-NO. SMP incubated in the presence of succinate showed an EPR signal (g=1.99) compatible with a stable semiquinone. This EPR signal was increased not only by antimycin but also by GSNO and SPER-NO. These signals were not modified under N2 atmosphere, indicating that they are not a consequence to the effect of NOx species on complex III area. These results show that NO interacts with ubiquinone-cytochrome b area producing antimycin-like effects. This behaviour comprises the inhibition of electron transfer, the interruption of the oxidation of cytochromes b, and the enhancement of [UQH(•)]ss which, in turn, leads to an increase in O2(•-) and H2O2 mitochondrial production rates.


Incidence of obstetric anal sphincter injuries following breech compared to cephalic vaginal births.
Obstetric anal sphincter injuries (OASIs) at the time of childbirth can lead to serious consequences including anal incontinence, dyspareunia, pain and rectovaginal fistula. These types of lesions and their incidence have been well studied after cephalic presentation deliveries, but no publications have specifically addressed this issue in the context of vaginal breech delivery. The goal of our study was to evaluate the incidence of OASIs following breech deliveries and compare it with cephalic presentation births. This was a retrospective cohort study involving 670 women. Of these, 224 and 446 had a vaginal birth of a fetus in the breech (breech group) and cephalic (cephalic group) presentations respectively. Both groups were matched for birthweight (± 200 g), date of delivery (± 2 years) and vaginal parity. Main outcome of interest was to evaluate the incidence of OASIs following breech vaginal birth compared to cephalic vaginal births. Secondary endpoints were the incidence of intact perineum or first-degree tear, second-degree perineal tear and rates of episiotomies in each group. There was no statistically significant difference in OASIs incidence between the breech and cephalic groups (0.9% vs. 1.1%; RR 0.802 (0.157; 4.101); p = 0.31). There were more episiotomies in the breech group (12.5% vs. 5.4%, p = 0.0012) and the rate of intact or first-degree perineum was similar in both groups (74.1% vs. 75.3%, p = 0.7291). A sub-analysis excluding patients with episiotomy and history of OASIs did not show any statistically significant difference either. We did not demonstrate a significant difference in the incidence of obstetric anal sphincter injuries between women who had a breech vaginal birth compared to cephalic.


CSF levels of HVA and 5-HIAA in drug-free schizophrenic patients and healthy controls: a prospective study focused on their predictive value for outcome in schizophrenia.
The CSF levels of HVA and 5-HIAA were determined in 90 drug-free DSM-III-R schizophrenic patients and 47 healthy control subjects, and their predictive value for 5-year outcome was evaluated. CSF was collected by lumbar puncture at index admission, and in 37 of the patients a second sample was drawn after approx. 7 weeks of neuroleptic treatment. Outcome was rated prospectively 5 years after index admission by means of the Strauss-Carpenter outcome scale. Schizophrenic patients had significantly lower levels of HVA in the CSF than the control group, but no difference was found for 5-HIAA. The CSF-amine metabolite levels were not correlated with age at admission, age at first symptoms or duration of the disorder. Neither HVA nor 5-HIAA correlated with the total outcome scores at a 1- and 5-year follow-up evaluation. First-admitted previously untreated patients with the poorest 5-year outcome had significantly lower HVA/5-HIAA quotients than those with a good outcome. Furthermore, patients still having a low HVA/5-HIAA quotient after treatment with neuroleptics had a poorer 5-year outcome than patients with an increased quotient. The data indicate that both HVA and 5-HIAA in the CSF, and especially their sensitivity to neuroleptic treatment, have a predictive value for the prognosis in schizophrenia.


The effect of aging on the growth of colon cancer.
Reports on the effect of patient age on the prognosis for colon cancer vary. We have tested the effect of aging using a model of murine colon adenocarcinoma in groups of mice of different ages. In experiment A, Balb/c mice of age groups 3-4 weeks, 10-12 weeks, 24-32 weeks and 40-48 weeks (13 mice/group) were injected with 5 X 10(4) MC-26 cells subcutaneously in the right flank. Tumors were measured twice weekly, and the rate of occurrence of tumor, mortality rate, and growth rate were calculated. In experiment B, the same plan as experiment A was used, except mice of age groups I (14 days), II (3-4 weeks), and III (20-22 months) were used with tumor doses of 1 X 10(4) cells and 5 X 10(5) cells (9-15 mice/group). In both experiments, the rate of growth of tumor, mortality rate, and sizes of tumors obtained were the same. In experiment A, the rate of occurrence of the tumors was the same in all groups, but in experiment B the occurrence of the tumor varied. A palpable tumor appeared earliest in the weanling mice (14 days), next in old mice (20-22 months), and last in the young adult group (3-4 weeks). Tumor doubling time was longest in the young adult mice (7 days), intermediate in the old mice (6.1 days), and shortest in the weanling mice (5.5 days). Established tumors grew at similar rates (as assessed by doubling time), independent of host age. Mortality rates were similar.(ABSTRACT TRUNCATED AT 250 WORDS)


Operational performance and analysis of two rabies vaccination campaigns in N'Djamena, Chad.
Transmission of rabies from animals to people continues despite availability of good vaccines for both human and animal use. The only effective strategy to achieve elimination of dog rabies and the related human exposure is to immunize dogs at high coverage levels. We present the analysis of two consecutive parenteral dog mass vaccination campaigns conducted in N'Djamena in 2012 and 2013 to advocate the feasibility and effectiveness for rabies control through proof of concept. The overall coverage reached by the intervention was >70% in both years. Monthly reported rabies cases in dogs decreased by more than 90% within one year. Key points were a cooperative collaboration between the three partner institutions involved in the control program, sufficient information and communication strategy to access local leaders and the public, careful planning of the practical implementation phase and the effective motivation of staff. The dynamic and semi to non-restricted nature of dog populations in most rabies endemic areas is often considered to be a major obstacle to achieve sufficient vaccination coverage. However, we show that feasibility of dog mass vaccination is highly dependent on human determinants of dog population accessibility and the disease awareness of dog owners. Consequently, prior evaluation of the human cultural and socio-economic context is an important prerequisite for planning dog rabies vaccination campaigns.


Protective effect of dexmedetomidine infusion combined with epidural blockade on postoperative complications after surgery: A prospective randomized controlled clinical trial.
This prospective, randomized, controlled study aimed to explore the efficacy of dexmedetomidine combined with epidural blockade on postoperative recovery of elderly patients after radical resection for colorectal cancer. Ninety-six elderly patients who underwent radical resection for colorectal cancer were randomly divided into the following four groups: dexmedetomidine, epidural blockade (ropivacaine), combined (dexmedetomidine + epidural blockade), and control (0.9% saline). The Mini-Mental State Examination (MMSE), Visual Analog Scale (VAS), and Ramsay scores at 48 hours, and time to first activity, length of hospital stay, and postoperative complication rates at 3 months were assessed. Twelve hours after surgery, Ramsay scores were higher in the combined compared with the control and epidural blockade groups. Twenty-four hours after surgery, MMSE scores were higher in the combined compared with the other groups. The combined group showed the lowest VAS scores except at 48 hours. Time to first activity and length of hospital stay were significantly shorter in the combined compared with the other groups. There was no difference in total postoperative complication rates among the groups. A combination of intraoperative dexmedetomidine infusion and epidural blockade could mitigate pain after surgery, improve cognitive dysfunction in early surgery, and facilitate recovery.


The investigation of the effect of thermal treatment on bentonites from Turkey with Fourier transform infrared and solid state nuclear magnetic resonance spectroscopic methods.
There is a great deal of interest in the building industry in burned clays for production of building materials. Therefore, the effect of heat treatment on natural bentonite from Turkey was investigated by Fourier transform infrared (FT-IR) between the region of 4000-400cm(-1) and (29)Si, (27)Al magic angle spinning nuclear magnetic resonance (MAS NMR) measurement techniques at various temperatures between 200 and 700°C for 2h. The structural changes were also investigated upon heat treatment.


In vivo and in vitro studies on apoptosis in OSE cells and inclusion cysts of pregnant heifers.
Elevated progesterone concentration during pregnancy and use of progesterone-like contraceptives are known to reduce ovarian cancers. This study was undertaken to decipher whether or not there is any relationship between progesterone (also oestrogen)-mediated ovarian surface epithelium (OSE) apoptosis and expression of p53, a cell-cycle arresting protein and potential tumour suppressor. Immunohistochemical staining with cytokeratin confirmed epithelial nature of the cells in the OSE layer and inclusion cysts that invaginate inside stroma after ovulation takes place. The in situ apoptosis index was determined during oestrus, and at mid and late-pregnancy stages in heifers. Epithelia of both tissues exhibited significantly high nuclear staining, suggesting that these cells are aiming to apoptotic destruction. To further establish a role of progesterone, the OSE cells were exposed in vitro to two concentrations of oestrogen and progesterone. It was revealed that progesterone at both concentrations and oestrogen only at high concentration converted a large proportion of these cells apoptotic. The stimulatory effect of progesterone (and to some extent oestrogen) was also seen on p53 expression in the same cultivated OSE cells. The steroid dosage dependence for apoptosis and p53 expression was also somewhat similar. Assuming that progesterone action is mediated through p53-caused apoptosis as a mechanism to evade malignant transformation of OSE cells, p53 expression at mRNA and protein level was investigated in the OSE layer in proximity to stroma, antrum and corpus luteum (CL). In cycling animals CL produces a large amount of progesterone and also oestrogen to maintain the post-ovulatory cycle and to suppress the gonadotropin production. Hence, cells undergoing re-epithelialization and which are in contact with CL were expected to undergo maximum apoptotic modification. Indeed we got the maximum p53/p53 gene expression in these cells. We conclude that progesterone during cycling and pregnancy may reduce the risk of developing ovarian cancer by ceasing cell cycle and diverting damaged and mutagenized OSE cells for apoptosis, and the process may be mediated through elevated p53 synthesis. However, it is also possible that progesterone and p53-induced apoptosis may be entirely different cancer suppressive actions but coincidently happening together.


Questioning the "big assumptions". Part I: addressing personal contradictions that impede professional development.
The ultimate success of recent medical curriculum reforms is, in large part, dependent upon the faculty's ability to adopt and sustain new attitudes and behaviors. However, like many New Year's resolutions, sincere intent to change may be short lived and followed by a discouraging return to old behaviors. Failure to sustain the initial resolve to change can be misinterpreted as a lack of commitment to one's original goals and eventually lead to greater effort expended in rationalizing the status quo rather than changing it. The present article outlines how a transformative process that has proven to be effective in managing personal change, Questioning the Big Assumptions, was successfully used in an international faculty development program for medical educators to enhance individual personal satisfaction and professional effectiveness. This process systematically encouraged participants to explore and proactively address currently operative mechanisms that could stall their attempts to change at the professional level. The applications of the Big Assumptions process in faculty development helped individuals to recognize and subsequently utilize unchallenged and deep rooted personal beliefs to overcome unconscious resistance to change. This approach systematically led participants away from circular griping about what was not right in their current situation to identifying the actions that they needed to take to realize their individual goals. By thoughtful testing of personal Big Assumptions, participants designed behavioral changes that could be broadly supported and, most importantly, sustained.


The addition of evolocumab to maximal tolerated statin therapy improves walking performance in patients with peripheral arterial disease and intermittent claudication (Evol-PAD study).
To test the hypothesis that in patients with peripheral arterial disease (PAD) and claudication, treated with maximal tolerated statin therapy, the addition of a monthly subcutaneous injection of evolocumab for 6 months improves treadmill walking performance. Lipid lowering therapy improves walking parameters in patients with PAD and claudication. Evolocumab decreases cardiac and limb adverse events in patients with PAD; however, the effect of evolocumab on walking performance is not known. We performed a double-blind, randomized, placebo-controlled study to compare maximal walking time (MWT) and pain free walking time (PFWT) in patients with PAD and claudication treated with monthly subcutaneous injections of evolocumab 420 mg (n = 35) or placebo (n = 35). We also performed measurements of lower limb perfusion, brachial flow mediated dilatation (FMD), carotid intima media thickness (IMT), and serum biomarkers of PAD disease severity. After six-months of treatment with evolocumab MWT increased by 37.7 % (87.5 ± 24 s) compared to 1.4 % (-21.7 ± 22.9 s) in the placebo group, p = 0.01. PFWT increased by 55.3 % (67.3 ± 21.2 s) in the evolocumab group compared to 20.3 % (8.5 ± 20.3 s) in the placebo group, p = 0.051. There was no difference in lower extremity arterial perfusion measurements. FMD increased by 42.0 ± 73.9 % (1.01 ± 0.7 %) in the evolocumab group and decreased by 16.29 ± 20.06 % (0.99 ± 0.68 %) in the placebo group (p < 0.001). IMT decreased by 7.16 ± 4.6 % (0.06 ± 0.04 mm) in the evolocumab group and increased by 6.68 ± 4.9 % (0.05 ± 0.03 mm) in the placebo group, (p < 0.001). The addition of evolocumab to maximal tolerated statin therapy improves maximal walking time in patients with PAD and claudication, increases FMD, and decreases IMT. Peripheral arterial disease (PAD) impairs quality of life by causing lower extremity intermittent claudication, rest pain, or amputation. Evolocumab is a monthly injectable monoclonal antibody medication that reduces cholesterol. In this study, we randomly treated patients with PAD and claudication, and on background statin therapy, with evolocumab or placebo, and found that evolocumab improves walking performance on a treadmill test by increasing maximal walking time. We also found that evolocumab decreases plasma MRP-14 levels, a marker of PAD severity.


High-nitrate wastewater treatment in an expanded granular sludge bed reactor and microbial diversity using 454 pyrosequencing analysis.
Denitrification of high concentration of nitrate wastewater was investigated in expanded granular sludge bed (EGSB) reactor with sodium acetate as the carbon source. The optimal parameters were achieved with C/N mole ratio of 2.0, liquid up-flow velocity (Vup) of 3.0 m/h and pH of 6.2-8.2. Complete denitrification can be achieved even with nitrate nitrogen concentration as high as 14000 mg/L. Furthermore, 454-pyrosequencing technology was used to analyze bacterial diversity. Results showed that a total of 5573 sequences were obtained which could be affiliated to 6 phylogenetic groups, including Proteobacteria, Firmicutes, Actinobacteria, Bacteroidetes, Chloroflexi and unclassified phylum. Proteobacteria (84.53%) was the dominant microbial population, followed by Firmicutes (13.24%) and Actinobacteria (0.38%). The dominate phylum was different from that in other anaerobic system.


Detection of Epstein-Barr virus transcripts in chemically or immunologically-activated cells and in a null cell-line (HLN-STL-C) by in situ hybridization with alkaline phosphatase-linked oligonucleotide probes.
We report a simple procedure for the detection of Epstein-Barr virus (EBV) by in situ DNA-RNA hybridization with an alkaline phosphatase-linked oligonucleotide probe. EBV-producing cell lines P3HR-1 and Akata were treated with phorbol ester and n-butyrate, and anti-human IgG, respectively. This treatment resulted in highly increased populations of cells with EBV transcripts of the latent membrane protein 1 (LMP1) and envelop glycoprotein gp350/220, but not of EBV-encoded small nuclear RNAs (EBERs). Synthesis of the LMP1 protein, which was encoded by the induced mRNA, was mostly dependent on viral DNA synthesis, as shown by double or single labeling for in situ DNA-DNA hybridization with the oligo-nucleotide probe, and immunoperoxidase staining with a monoclonal antibody against LMP1. In situ hybridization of the null cell line HLN-STL-C established from an adult T-cell leukemia patient showed that 100% of the cells contained both EBERs and LMP1 mRNA and about 0.1% of the cells contained gp350/220 mRNA, indicating that a few of the null cells which carried the EBV genome spontaneously entered the late EBV replication cycle.


Postmenopausal hormone use and cholecystectomy in a large prospective study.
To examine the association between postmenopausal hormone use and cholecystectomy. A prospective cohort study was performed, with follow-up every 2 years. Participants were 54,845 postmenopausal United States nurses, who reported both hormone use and cholecystectomy on mailed questionnaires. Cholecystectomy was reported by 1750 women during 8 years of follow-up. After adjusting for confounding factors, women currently using postmenopausal hormones were at an increased risk of cholecystectomy (relative risk [RR] 2.1, 95% confidence interval [CI] 1.9-2.4) compared to never-users. For current users, the risk of cholecystectomy increased with increasing duration of hormone use (RR 2.6, 95% CI 2.2-3.1 for 10 years or more) and higher doses of estrogen (RR 2.4, 95% CI 2.0-2.9 for users of 1.25 mg or more). Although the risk for past hormone users decreased substantially in women who had discontinued use 1-2.9 years ago (RR 1.6, 95% CI 1.2-2.0), a small risk persisted for women who had stopped taking hormones 5 or more years previously (RR 1.3, 95% CI 1.1-1.6). However, after controlling for time since last use, duration of past use had little or no effect on the risk of cholecystectomy (RR 1.4 and RR 1.7 for past users of less than 2 years and 10 or more years' duration, respectively). Women using postmenopausal hormones are at an increased risk of cholecystectomy. Women and their physicians should consider the spectrum of risks and benefits when deciding whether to take hormones.


Acute phase glycoproteins in sera of patients with sarcomas receiving methotrexate infusion therapy.
The serum concentrations of alpha-1-acid glycoprotein, haptoglobin and alpha-1-antitrypsin have been estimated serially in eight patients with malignant tumours of bone and soft tissue, who received a total of thirty-four intravenous infusions of high-dose methotrexate. The serum glycoprotein levels taken before the first drug exposure did not relate with the prognosis of these patients. A tumour mass (40 mm) did not influence the serum levels of these glycoproteins. The presence of a pleural effusion was associated with increased serum levels of these glycoproteins. These assays appear to be of no value in monitoring the course of the disease in this group of cancer patients.


Increasing adult Tdap vaccination rates by vaccinating infant caregivers in the pediatric office.
To increase adult caregiver Tdap vaccination rates by offering Tdap vaccine during infant well-child visits. We developed a pilot vaccine initiative wherein pediatricians offered Tdap vaccine to mothers and non-mother caregivers attending the 2-week well-child visit at a hospital-based clinic serving predominantly low-income families. We evaluated this initiative by asking mothers and caregivers to participate in a survey after the 2-week visit to determine self-reported Tdap vaccination status, demographics, and the source of their adult primary care. Seventy (69%) participants received the Tdap vaccine during the newborns' 2-week well-child visit. Forty-six percent of the infants' 152 adult household contacts were vaccinated through this initiative. Of those mothers and caregivers, more caregivers reported not having insurance (38% vs 15%, P < .001), and no routine medical care (23% vs 8%, P = .007). Through this pilot initiative, we vaccinated 69% of mothers and non-mother caregivers presenting to the 2-week well-child visit. A large proportion of caregivers did not receive routine medical care or have insurance, which suggests that they otherwise may have poor access to the vaccine. Tdap vaccination in the pediatric office represents a substantial opportunity to increase vaccination rates.


The involvement of protein kinase C and tyrosine kinase in vanadate-induced contraction.
Gastric smooth muscle of cats was used to investigate the involvement of protein kinase in vanadate-induced contraction. Vanadate caused a contraction of cat gastric smooth muscle in a dose-dependent manner. Vanadate-induced contraction was totally inhibited by 2 mM EGTA and 1.5 mM LaCl3 and significantly inhibited by 10 microM verapamil and 1 microM nifedipine, suggesting that vanadate-induced contraction is dependent on the extracellular Ca2+ concentration. and the influx of extracellular Ca2+ was mediated through voltage-dependent Ca2+ channel. Both protein kinase C inhibitor and tyrosine kinase inhibitor significantly inhibited the vanadate-induced contraction and the combined inhibitory effect of two protein kinase inhibitors was greater than that of each one. But calmodulin antagonists did not have any influence on the vanadate-induced contraction. On the other hand, both forskolin (1 microM) and sodium nitroprusside (1 microM) significantly inhibited vanadate-induced contraction. Therefore, these results suggest that both protein kinase C and tyrosine kinase are involved in the vanadate-induced contraction which required the influx of extracellular Ca2+ in cat gastric smooth muscle, and that the contractile mechanism of vanadate may be different from that of agonist binding to its specific receptor.


Implications of expansin-like 3 gene in Dictyostelium morphogenesis.
Dictyostelium harbors multiple expansin-like genes with generally unknown functions. Thus, we analyzed the expansin-like 3 (expL3) gene and found that its expression was reduced in a null mutant for a STATa gene encoding a transcription factor. The expression of expL3 was developmentally regulated and its transcript was spliced only in the multicellular stages. The expL3 promoter was activated in the anterior prestalk region of the parental strain and downregulated in the STATa null slug, although the expL3 promoter was still expressed in the prestalk region. The expL3 overexpressing strain exhibited delayed development and occasionally formed an aberrant structure, i.e., a fruiting body-like structure with a short stalk. The ExpL3-myc protein bound cellulose.


Circular RNA CircDHRS3 Aggravates IL-1β-induced ECM Degradation, Apoptosis, and Inflammatory Response via Mediating MECP2 Expression.
Previous studies have reported that circular RNA hsa_circ_0010024 (circDHRS3), microRNA (miR)-193a-3p, and Methyl CpG binding protein 2 (MECP2) are unconventionally expressed in osteoarthritis (OA) cartilage samples. However, the regulatory mechanisms among circDHRS3, miR-193a-3p, and MECP2 in OA pathogenesis are unclear. Changes of circDHRS3, miR-193a-3p, and MECP2 mRNA were detected by qRT-PCR. Several protein levels were evaluated using western blotting. Cell proliferation was analyzed by 5-Ethynyl-2'-deoxyuridine (EdU) and cell counting assays. Cell apoptosis was determined by flow cytometry assay. Detection of pro-inflammatory cytokines was conducted using ELISA. The relationship between circDHRS3 or MECP2 and miR-193a-3p was validated by dual-luciferase reporter assay. We verified that circDHRS3 and MECP2 were overexpressed in OA cartilage samples, whereas miR-193a-3p was downregulated. CircDHRS3 silencing weakened IL-1β-induced chondrocyte cartilage extracellular matrix (ECM) degradation, apoptosis, and inflammatory response. CircDHRS3 adsorbed miR-193a-3p to modulate MECP2 expression. Also, silencing of miR-193a-3p impaired circDHRS3 silencing-mediated suppression on IL-1β-induced chondrocyte injury. Also, MECP2 overexpression alleviated miR-193a-3p mimic-mediated inhibition on IL-1β-prompted chondrocyte injury. CircDHRS3 silencing reduced MECP2 expression via sponging miR-193a-3p, thereby weakening IL-1β-induced chondrocyte ECM degradation, apoptosis, and inflammatory response.


Functional Fe3O4@ZnO magnetic nanoparticle-assisted enrichment and enzymatic digestion of phosphoproteins from saliva.
Saliva contains various proteins, particularly abundant are phosphoproteins, that may be related to disease occurrences and that play significant roles in a biological system. Thus, medical diagnostics will benefit tremendously if disease-related protein biomarkers are discovered from saliva. In this paper, we propose and demonstrate an approach using functional zinc oxide coated iron oxide magnetic nanoparticles (Fe(3)O(4)@ZnO MNPs) as affinity probes to selectively enrich phosphoproteins from complex saliva samples and as microwave absorbers to assist the enrichment and subsequent tryptic digestion of trapped proteins under microwave heating. The target species trapped by MNPs were characterized by matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) combined with protein database search. Entire analysis time was shortened to less than 20 min. The detection limit of this approach for a monophosphopeptide was as low as 250 pM (10 μL).


Autophagy is inhibited by ubiquitin ligase activity in the nervous system.
Autophagy is an intracellular catabolic process prominent in starvation, aging and disease. Neuronal autophagy is particularly important, as it affects the development and function of the nervous system, and is heavily implicated in neurodegenerative disease. Nonetheless, how autophagy is regulated in neurons remains poorly understood. Using an unbiased proteomics approach, we demonstrate that the primary initiator of autophagy, the UNC-51/ULK kinase, is negatively regulated by the ubiquitin ligase RPM-1. RPM-1 ubiquitin ligase activity restricts UNC-51 and autophagosome formation within specific axonal compartments, and exerts effects broadly across the nervous system. By restraining UNC-51 activity, RPM-1 inhibits autophagosome formation to affect axon termination, synapse maintenance and behavioral habituation. These results demonstrate how UNC-51 and autophagy are regulated subcellularly in axons, and unveils a mechanism for restricting initiation of autophagy across the nervous system. Our findings have important implications beyond nervous system development, given growing links between altered autophagy regulation and neurodegenerative diseases.


Parity, other reproductive factors, and risk of pancreatic cancer mortality in a large cohort of U.S. women (United States).
Several studies have found an inverse relationship between parity and risk of pancreatic cancer. However, most of the studies of pancreatic cancer in relation to parity and other reproductive factors have been small and the results inconsistent. Most recently, a well-conducted, prospective cohort study found a linear inverse association between parity and pancreatic cancer. To clarify the relationship between parity and other reproductive factors and risk of pancreatic mortality, we examined these associations among 387,981 postmenopausal U.S. women in the Cancer Prevention Study (CPS)-II cohort. CPS-II participants completed a self-administered questionnaire in 1982 and were followed for mortality through 2000. During follow-up, 1959 pancreatic cancer deaths occurred. Using Cox proportional hazards modeling, we calculated rate ratios (RR) adjusted for age, race, education, personal history of diabetes, body mass index, height, exercise, family history of pancreatic cancer, and cigarette smoking status, frequency, and duration. Overall, we did not observe a significant association between parity and pancreatic cancer mortality (trend p = 0.07). However, women who had five or more births had lower death rates from pancreatic cancer than nulliparous women (RR = 0.80, 95% CI = 0.66-0.96). We observed no association between any other reproductive factors examined (age at first birth, menarche, or menopause; type of menopause; diethylstilbestrol (DES) use; or duration of oral contraceptive or estrogen replacement therapy use) and pancreatic cancer mortality. In summary, our results support the observation that high parity is associated with lower risk of pancreatic cancer but do not show a linear trend with increasing parity. Furthermore, we find no evidence that other reproductive factors are associated with pancreatic cancer mortality.


Development and validation of fear of hypoglycemia screener: results from the T1D exchange registry.
Fear of Hypoglycemia (FoH) in people with diabetes has a significant impact on their quality of life, psychological well-being, and self-management of disease. There are a few questionnaires assessing FoH in people living with diabetes, but they are more often used in research than clinical practice. This study aimed to develop and validate a short and actionable FoH screener for adults living with type 1 diabetes (T1D) for use in routine clinical practice. We developed an initial screener based on literature review and, interviews with healthcare providers (HCPs) and people with T1D. We developed a cross-sectional web-based survey, which was then conducted to examine the reliability and validity of the screener. Adults (aged ≥ 18 years) with diagnosis of T1D for ≥ 1 year were recruited from the T1D Exchange Registry (August-September 2020). The validation analyses were conducted using exploratory factor analyses, correlation, and multivariable regression models for predicting cut-off scores for the final screener. The final FoH screener comprised nine items assessing two domains, "worry" (6-items) and "avoidance behavior" (three items), in 592 participants. The FoH screener showed good internal consistency (Cronbach's α = 0.88). The screener also demonstrated high correlations (r = 0.71-0.75) with the Hypoglycemia Fear Survey and moderate correlations with depression, anxiety, and diabetes distress scales (r = 0.44-0.66). Multivariable regression analysis showed that higher FoH screener scores were significantly associated with higher glycated hemoglobin (HbA1c) (b = 0.04) and number of comorbidities (b = 0.03). This short FoH screener demonstrated good reliability and validity. Further research is planned to assess clinical usability to identify patients with FoH and assist effective HCP-patient conversations.


Diazepam dissociates the analgesic and aversive effects of periaqueductal gray stimulation in the rat.
Electrical stimulation of the periaqueductal gray matter (PAG) of the rat can produce both analgesia and aversive reactions. To determine if these two effects can be dissociated, diazepam, a benzodiazepine, was administered to rats chronically implanted with electrodes in the PAG. The threshold for stimulation-produced analgesia or aversion, whichever was lowest, was determined before and after drug administration. Diazepam (1 mg/kg) attenuated stimulation-produced aversive reactions at 12 of 20 stimulation sites, allowing analgesia to be measured at the same threshold. Diazepam did not alter baseline pain sensitivity or thresholds for stimulation-produced analgesia. These results indicate that aversive reactions and analgesia from PAG stimulation can be pharmacologically dissociated.


Imaging Chambers for Arabidopsis Seedlings for Mitotic Studies.
Flowering plants evolved away from creating centrosomes or conventional microtubule organizing centers. Therein, plants have posed a long-standing challenge to many of the conventional ideas for mitotic spindle construction and the process of chromosome segregation. The Arabidopsis seedling has emerged as a leading model for plant cell biological studies of the cytoskeleton and vesicle trafficking. Here we describe methods for creating a reusable chamber for mitotic studies in both seedling root and shoot cells with instruction for best practices with conventional microscopic techniques.


Pregnancy affects appraisal of negative life events.
It has been demonstrated that physiological responses to stress are diminished late in pregnancy. This study investigates whether emotional responding is diminished as well by measuring affective responses to specific life events during pregnancy. A total of 292 pregnant women reported the occurrence of and affective responses to a range of major life events during gestation. Two analyses were conducted (across events and within events) on these responses to determine whether life events occurring in the first trimester were rated as more stressful than those that occurred in the third trimester. Both within-event and across-events analyses of responses to life events demonstrated that events occurring early in pregnancy were perceived as more stressful than events occurring later in pregnancy. Responses to stress and affective state are progressively altered in pregnant women, suggesting that timing of stress exposure during gestation may be critical in determining its impact.


Modified Sandwich-graft Technique Employing Aorfix and Viabahn Stent-grafts to Preserve Hypogastric Flow in Cases of Complex Aortoiliac and Isolated Common Iliac Artery Aneurysms Including the Internal Iliac Artery Ostium.
The aim was to assess the early and mid-term safety and clinical outcomes of a modified sandwich-graft technique (MSGT) that employed the Aorfix and Viabahn stent-grafts to preserve hypogastric flow in cases of complex aortoiliac and isolated common iliac artery (CIA) aneurysms including internal iliac artery (IIA) ostium who were not suitable for an iliac branch device (IBD). A review of prospective collected data of all consecutive cases of treatment of complex aortoiliac or isolated CIA aneurysms including IIA ostium using the MSGT to preserve the hypogastric flow in three European centres between April 1, 2010, and December 31, 2013, was performed. All patients included were unfit for open repair and not suitable for an IBD. During the study period, 21 patients met the study criteria and had 25 MSGTs. The mean follow-up duration was 17.2 months. The technical success rate was 100%, with no perioperative mortality and stent-graft or MSGT-treated IIA occlusions. There were two cases with Type Ib endoleaks and one Type III endoleaks intraoperatively that were successfully managed. There was also no reported early and late pelvic ischaemia. The primary patency rate was 90.5%, and two cases of MGST treated IIA occlusions with no reported symptoms of pelvic ischaemia during follow-up. MSGT was a safe and feasible strategy to preserve hypogastric flow in cases of complex aortoiliac and isolated CIA aneurysms including IIA ostium and who were not suitable for IBD, with encouraging early and mid-term outcomes.


Clinical results after single asymmetrical shark fin for graft orientation in DMEK.
Evaluating the effect of a single peripheral triangular mark to ensure the correct anterior-posterior graft orientation in DMEK. Retrospective study of patients scheduled for DMEK due to Fuchs endothelial dystrophy and divided into 2 study groups: Group -M (n = 184) had no mark of the EDM (Endothelial Descemet membrane) and group + M (n = 193) had a triangular peripheral mark. Follow-up time was 1 year after surgery. The postoperative graft turning and Re-DMEK rate could be significantly reduced by the use of a peripheral mark (p = 0.002, p = 0.001, respectively). Re-DMEK due to primary graft failure was significantly associated with prior graft turning (p < 0.001). Both groups showed comparable values for visual acuity, central corneal thickness and endothelial cell count after a follow-up of 1 year. Single peripheral triangular marking is a simple and cost-saving addition to EDM preparation to ensure the correct orientation of the graft intraoperatively and could lead to a significant reduction in graft turning and re-DMEK rate in this study.


Managing disorders of consciousness: the role of electroencephalography.
Disorders of consciousness (DOC) are an important but still underexplored entity in neurology. Novel electroencephalography (EEG) measures are currently being employed for improving diagnostic classification, estimating prognosis and supporting medicolegal decision-making in DOC patients. However, complex recording protocols, a confusing variety of EEG measures, and complicated analysis algorithms create roadblocks against broad application. We conducted a systematic review based on English-language studies in PubMed, Medline and Web of Science databases. The review structures the available knowledge based on EEG measures and analysis principles, and aims at promoting its translation into clinical management of DOC patients.


Quantification of airborne African swine fever virus after experimental infection.
Knowledge on African Swine Fever (ASF) transmission routes can be useful when designing control measures against the spread of ASF virus (ASFV). Few studies have focused on the airborne transmission route, and until now no data has been available on quantities of ASF virus (ASFV) in the air. Our aim was to validate an air sampling technique for ASF virus (ASFV) that could be used to detect and quantify virus excreted in the air after experimental infection of pigs. In an animal experiment with the Brazil'78, the Malta'78 and Netherlands'86 isolates, air samples were collected at several time points. For validation of the air sampling technique, ASFV was aerosolised in an isolator, and air samples were obtained using the MD8 air scan device, which was shown to be suitable to detect ASFV. The half-life of ASFV in the air was on average 19 min when analysed by PCR, and on average 14 min when analysed by virus titration. In rooms with infected pigs, viral DNA with titres up to 10(3.2) median tissue culture infective dose equivalents (TCID50eq.)/m(3) could be detected in air samples from day 4 post-inoculation (dpi 4) until the end of the experiments, at dpi 70. In conclusion, this study shows that pigs infected with ASFV will excrete virus in the air, particularly during acute disease. This study provides the first available parameters to model airborne transmission of ASFV.


Development of a resilience scale for Thai substance-dependent women: A mixed methods approach.
The purpose of this study was to develop a resilience scale based on the experiences of substance-dependent women in Thailand and evaluate its validity and reliability. A sequential exploratory mixed methods design was employed as the main methodology to develop the resilience scale according to the results from qualitative data by analyzing focus group discussions of 13 participants. Then, the scale was administered to 252 substance-dependent women from four substance-treatment centers. The psychometric properties were explored with an index of item objective congruence (IOC), Pearson correlation, second-order confirmatory factor analysis and Cronbach's alpha coefficient to estimate the quantitative data. The qualitative results showed that resilience is defined by three themes: individual, family and community factors, consisted of 13 different categories. The quantitative results also revealed that all 71 items in the resilience scale passed the IOC criteria, convergence and construct validity. The goodness-of-fit indices demonstrated that the resilience model was consistent with the empirical data. (Chi-square=74.28, df=59, p-value=0.08, RMSEA=0.03, SRMR=0.04, NNFI=0.99, CFI=0.99, GFI=0.96). The internal consistency, assessed by a Cronbach's alpha score of 0.92, can be interpreted as demonstrating high reliability. Furthermore, the structure of the resilience scale was confirmed by the available resilience literature. This study can help clinicians gain a more comprehensive understanding regarding the complex process of resilience among substance-dependent women and aid them in providing these women with the appropriate interventions.


Hydrochemical and geoelectrical investigation of the coastal shallow aquifers in El-Omayed area, Egypt.
Monitoring and assessment of the coastal aquifers are becoming a worldwide concern for the need of additional and sustainable water resources to satisfy demographic growth and economic development. A hydrochemical and geoelectrical investigation was conducted in the El-Omayed area in the northwestern coast of Egypt. The aim of the study was to delineate different water-bearing formations, provide a general evaluation of groundwater quality, and identify the recharge sources in aquifers. Thirty-seven water samples were collected and chemically analyzed from the sand dune accumulations and oolitic limestone aquifers. Fifteen profiles of vertical electrical soundings (VESs) were obtained in the oolitic limestone aquifer to examine the variations of subsurface geology and associated groundwater chemistry. The groundwater reserves in the El-Omayed area are mainly contained in sand dune accumulations and oolitic limestone aquifers. The aquifer of sand dune accumulations contains freshwater of low salinity (average total dissolved solids (TDS) = 974 mg/l). Groundwater of oolitic limestone aquifer is slightly brackish (average TDS = 1,486 mg/l). Groundwater of these aquifers can be used for irrigation under special management for salinity control, and regular leaching as indicated by electrical conductivity and sodium adsorption ratio. Results of VES interpretation classified the subsurface sequence of oolitic limestone aquifer into four geoelectric zones, with increasing depth, calcareous loam, gypsum, oolitic limestone, and sandy limestone. Oolitic limestone constitutes the main aquifer and has a thickness of 12-32 m.


Dysrhythmia update. Differential diagnosis of supraventricular tachycardia in an elderly man.
An 82-year-old man was admitted to the hospital with a narrow complex tachycardia. This article presents his case history, examples of the dysrhythmia, and a differential diagnosis approach to identifying the source of the tachycardia. The mechanism of the tachycardia is described and diagrammed and the follow-up care of the patient presented.


Robustness to source displacement in dual air kerma strength planning for focal low-dose-rate brachytherapy of prostate cancer.
To describe the use of dual source strength implants for focal low-dose-rate brachytherapy. An interneedle dual source strength planning strategy is described for focal low-dose-rate brachytherapy of the prostate. The implanted treatment plans were designed using peripheral (except near the rectum) needles loaded with high strength (0.9 U) sources and central needles loaded with low strength (0.4 U) sources ("interneedle" dual strength planning). This approach has been applied for focally treating 3 patients. In this article, we compare the characteristics and robustness to source motion of interneedle dual strength planning with four alternative planning strategies (single strength high, low, and intermediate, and intraneedle dual strength) on 50 simulated cases. Interneedle dual source strength planning results in greater robustness to source motion and overall lower seed and needle density compared to the standard low source strength planning currently used in our centre. This planning approach is also significantly superior to single strength high, single strength intermediate and intraneedle dual strength planning strategies in terms of high dose to the urethral avoidance structure. The use of interneedle dual source strength treatment plans for focal low-dose-rate brachytherapy is possibly the practical solution for limiting the density of sources required to deliver the prescribed dose while limiting proximity of high strength sources to organs at risk.


How does baseline anthropometry affect anthropometric outcomes in children receiving treatment for severe acute malnutrition? A secondary analysis of a randomized controlled trial.
Mid-upper arm circumference (MUAC) < 11.5 cm and weight-for-height Z-score (WHZ) < -3 are used for screening for severe acute malnutrition (SAM). Underweight and concurrent wasting and stunting may better target those at the highest risk of mortality. We compared anthropometric outcomes in children enrolled in a trial of antibiotics for SAM based on categories of baseline anthropometry, including indicators for programme admission (WHZ < -3, MUAC < 11.5) and alternative indicators (weight-for-age Z-score [WAZ] < -3, concurrent wasting and stunting [WHZ < -3 and height-for-age Z-score < -3]). Participants were followed weekly until nutritional recovery and at 8 weeks. We evaluated changes in weight gain (g/kg/day), MUAC, and WHZ in children admitted by admissions criteria (MUAC only, WHZ only, or MUAC and WHZ) and by underweight or concurrent wasting and stunting. Of 301 admitted children, 100 (33%) were admitted based on MUAC only, 41 (14%) WHZ only, and 160 (53%) both MUAC and WHZ, 210 (68%) were underweight and 67 (22%) were concurrently wasted/stunted. Low MUAC and low WHZ children had the lowest probability of nutritional recovery (17% vs. 50% for MUAC-only and 34% for WHZ-only). There was no difference in weight gain velocity or WHZ by admissions criteria (WHZ and/or MUAC). Underweight and concurrently wasted/stunted children had lower MUAC and WHZ at 8 weeks compared with those who were not underweight or concurrently wasted and stunted. Children with both low MUAC and low WHZ had the worst outcomes. Relying on MUAC alone may miss children who have poor outcomes. Other indicators, such as WAZ, may be useful for identifying vulnerable children.


Tenecteplase Improves Reperfusion across Time in Large Vessel Stroke.
Tenecteplase improves reperfusion compared to alteplase in patients with large vessel occlusions. To determine whether this improvement varies across the spectrum of thrombolytic agent to reperfusion assessment times, we performed a comparative analysis of tenecteplase and alteplase reperfusion rates. Patients with large vessel occlusion and treatment with thrombolysis were pooled from the Melbourne Stroke Registry, and the EXTEND-IA and EXTEND-IA TNK trials. The primary outcome, thrombolytic-induced reperfusion, was defined as the absence of retrievable thrombus or >50% reperfusion at imaging reassessment. We compared the treatment effect of tenecteplase and alteplase, accounting for thrombolytic to assessment exposure times, via Poisson modeling. We compared 90-day outcomes of patients who achieved reperfusion with a thrombolytic to patients who achieved reperfusion via endovascular therapy using ordinal logistic regression. Among 893 patients included in the primary analysis, thrombolytic-induced reperfusion was observed in 184 (21%) patients. Tenecteplase was associated with higher rates of reperfusion (adjusted incidence rate ratio [aIRR] = 1.50, 95% confidence interval [CI] = 1.09-2.07, p = 0.01). Findings were consistent in patient subgroups with first segment (aIRR = 1.41, 95% CI = 0.93-2.14) and second segment (aIRR = 2.07, 95% CI = 0.98-4.37) middle cerebral artery occlusions. Increased thrombolytic to reperfusion assessment times were associated with reperfusion (tenecteplase: adjusted risk ratio [aRR] = 1.08 per 15 minutes, 95% CI = 1.04-1.13 vs alteplase: aRR = 1.06 per 15 minutes, 95% CI = 1.00-1.13). No significant treatment-by-time interaction was observed (p = 0.87). Reperfusion via thrombolysis was associated with improved 90-day modified Rankin Scale scores (adjusted common odds ratio = 2.15, 95% CI = 1.54-3.01) compared to patients who achieved reperfusion following endovascular therapy. Tenecteplase, compared to alteplase, increases prethrombectomy reperfusion, regardless of the time from administration to reperfusion assessment. Prethrombectomy reperfusion is associated with better clinical outcomes. ANN NEUROL 2023;93:489-499.


Supplementation of nutritional fluid and electrolyte balance by perfusion of an isolated small bowel loop in a patient with Crohn's disease.
Delayed reanastamosis of small bowel in Crohn's disease has been made possible, largely by parenteral nutrition. We describe a patient with multiple resections resulting in loss of all the small and large bowel except the duodenum and 70 cms of proximal jejunum ending in a jejunostomy and a 150 cm long isolated loop of jejuno-ileum. The short proximal bowel proved adequate for energy and protein nutrition, with small frequent meals but gave rise to a persistent negative balance of water, sodium, potassium and magnesium. This was corrected by overnight perfusion of the isolated loop with an electrolyte solution with added glucose polymer to enhance sodium absorption. The critical rate of perfusion and concentration of glucose polymer were determined by a series of studies. The patient and her husband carried out this regimen for a year at home. The isolated loop was subsequently reanastamosed to the proximal jejunum allowing resumption of normal eating without further electrolyte disturbance.


Breast cancer metastasis alters acetylcholinesterase activity and the composition of enzyme forms in axillary lymph nodes.
Because of the probable involvement of cholinesterases (ChEs) in tumorigenesis, this research was addressed to ascertaining whether breast cancer metastasis alters the content of acetylcholinesterase (AChE) and/or butyrylcholinesterase (BuChE) in axillary lymph nodes (LN). ChE activity was assayed in nine normal (NLN) and seven metastasis-bearing nodes (MLN) from women. AChE and BuChE forms were characterised by sedimentation analyses, hydrophobic chromatography and western blotting. The origin of ChEs in LN was studied by lectin interaction. AChE activity dropped from 21.6 mU/mg (nmol of the substrate hydrolysed per minute and per milligram protein) in NLN to 3.8 mU/mg in MLN (p < 0.001), while BuChE activity (3.6 mU/mg) was little affected. NLN contained globular amphiphilic AChE dimers (G2A, 35%), monomers (G1A, 30%), hydrophilic tetramers (G4H, 8%), and asymmetric species (A4, 23%, and A8, 4%); MLN displayed only G2A (65%) and G1A (35%) AChE forms. NLN and MLN contained G4H (79%), G4A (7%), and G1H (14%) BuChE components. Neither the binding of ChE forms with lectins and antibodies nor the subunit size were altered by metastasis. The higher level of AChE in NLN than in brain and the specific pattern of AChE forms in NLN support its role in immunity. The different profile of AChE forms in NLN and MLN may be useful for diagnosis.


Multifactorial pathogenesis: ought we to classify disease or treat the individual's causes of disease?
Added to the uniqueness of most of us imparted by HLA, our individual differences in the expression of other genetic traits, foetal period steroid imprints on brain and other functions, and regulators shared by body and mind indicate that each one of us (except most monozygotic twins) is born as a unique individual whole (I). Mechanisms suspected to be contributing ones in classified synovitides appear to correspond to individually exaggerated or depressed modes of response by physiological mechanisms linked to our unique individual wholes (II). If we upgrade the importance of influences of combinations of 'contributing' mechanisms, the importance of the alleged unknown fundamental causes of these diseases diminishes (III). This broad interpretation of 'multifactorial pathogenesis' can probably be applied to many common types of disease (IV), implying that disease classifications may poorly reflect the individual's causes of disease (V). There is reason to challenge modern concepts about disease (VI).


Perspectives of Academic Oncologists About Offering Expanded Access to Investigational Drugs.
The expanded access (EA) pathway permits patients to be treated with investigational medical products outside clinical trials. Because cancer care is a common indication for which EA is sought and these efforts require physician management, understanding oncologists' perspectives can help illuminate factors influencing patient access. To learn how oncologists practicing at academic medical centers (AMCs) perceive EA and their role in offering it. This qualitative study used data from semistructured interviews conducted from February 2020 to September 2021 with a purposive sample of oncologists recruited from large, urban AMCs in the northeast United States. Oncologists who had submitted at least 1 single-patient EA request to the institutional review boards at the University of Pennsylvania, Children's Hospital of Philadelphia, NYU Langone Health, and Dana-Farber Cancer Institute from January 1, 2014, through January 31, 2020, were eligible to participate. Data were analyzed from July 2021 to March 2022. Interviews focused on oncologist practice demographics, experience with EA, factors relevant to decisions to pursue EA and comfort with those decisions, perspectives on oncologists' role in EA, perspectives on the FDA's role, and the Right to Try pathway to access investigational drugs. Eligible oncologists were interviewed until thematic saturation was reached, resulting in 25 interviews; most participants were women (15 participants [60%]), reported primarily treating adult patients (15 participants [60%]), had more than 10 years of clinical experience (16 participants [64%]), and had submitted at least 2 single-patient EA requests to their institutional review boards during the relevant period (14 participants [56%]). Oncologists viewed EA as an important tool for securing what they determined to be the best treatment option for their patients based on their own expert assessment of available data. Interviewees reported that they would rather access interventions as commercially available products or through clinical trials; however, if the preferred option was not available through these means, they viewed pursuit of EA as part of their obligation to patients, while often recognizing the potential for inequities in the broader patient population beyond their institutions. Participating oncologists felt confident pursuing investigational drugs for treatment use, despite the absence of FDA marketing approval, and did not necessarily view EA as a last resort. These findings indicate that oncologists practicing in large academic settings sought to treat patients with the interventions they deemed most likely to be beneficial, regardless of approval status. As such, they viewed EA as an unexceptional means to obtain promising products, although it remains unclear whether their confidence in evaluating investigational treatments was justified. Future research should examine whether oncologists outside large AMCs share this confidence, as differences may influence patient access to the EA treatment pathway.


Correction to: Biodegradable magnesium-based screw clinically equivalent to titanium screw in hallux valgus surgery: short term results of the first prospective, randomized, controlled clinical pilot study.
An amendment to this paper has been published and can be accessed via the original article.


Enzymatic biotransformation of ginsenoside Rb1 to 20(S)-Rg3 by recombinant β-glucosidase from Microbacterium esteraromaticum.
Microbacterium esteraromaticum was isolated from ginseng field. The β-glucosidase gene (bgp1) from M. esteraromaticum was cloned and expressed in Escherichia coli BL21 (DE3). The bgp1 gene consists of 2,496 bp encoding 831 amino acids which have homology to the glycosyl hydrolase family 3 protein domain. The recombinant β-glucosidase enzyme (Bgp1) was purified and characterized. The molecular mass of purified Bgp1 was 87.5 kDa, as determined by SDS-PAGE. Using 0.1 mg ml(-1) enzyme in 20 mM sodium phosphate buffer at 37°C and pH 7.0, 1.0 mg ml(-1) ginsenoside Rb1 was transformed into 0.444 mg ml(-1) ginsenoside Rg3 within 6 h. The Bgp1 sequentially hydrolyzed the outer and inner glucose attached to the C-20 position of ginsenosides Rb1. Bgp1 hydrolyzed the ginsenoside Rb1 along the following pathway: Rb1 → Rd → 20(S)-Rg3. This is the first report of the biotransformation of ginsenoside Rb1 to ginsenoside 20(S)-Rg3 using the recombinant β-glucosidase.


Management of nontraumatic vascular shunts involving the cavernous sinus.
The authors managed 38 consecutive cases of nontraumatic vascular shunts involving the cavernous sinus. Selective angiography demonstrated 12 carotid cavernous fistulas (CCFs) and 26 dural arteriovenous shunts (DAVSs). Visual disability occurred from glaucoma, venous retinopathy, optic neuropathy, or diplopia. Ten patients with slow-flow shunts and minimal dysfunction were treated medically to lower intraocular pressure (IOP) and/or instructed in manual compression of the internal carotid artery, ipsilateral to the lesion, using the contralateral hand. Percutaneous intraarterial embolization using detachable balloons, isobutylcyanoacrylate, or polyvinyl alcohol particles was successful in 16/18 DAVSs and 9/10 CCFs. The neuro-ophthalmic signs resolved in these 25 cases. Complications occurred in five patients. These included a transient hemiparesis, twelfth nerve palsy, unilateral nasal field loss, a pseudoaneurysm causing a third-nerve paresis, and temporary cavernous sinus thrombosis. Conservative therapy in mild cases and embolization in cases with visual disability or progressive signs are warranted.


Fate of human thyroid tissue autotransplants.
We conducted this study to establish whether human thyroid tissue autografts can survive and function in the absence of their native blood supply in muscle. The benefits of this potential could be incorporated in routine surgery to reduce the incidence of post-operative hypothyroidism. Fifteen patients with benign thyroid disorders, seven of whom had Graves' disease and eight, multinodular goiter (MNG), underwent modified subtotal thyroidectomy and the autotransplantation of thyroid tissue in the sternocleidomastoid muscle. About 3-5 g of thyroid tissue was cut and implanted into the sternocleidomastoid muscle. Postoperative clinical assessment, thyroid function tests, and technetium scans of the neck were done to assess the function of remnant and transplanted thyroid tissue. The transplanted tissue was functional in six of the eight patients with MNG and four of the seven with Graves' disease. All the patients with MNG and a functional transplant became euthyroid within 6 months postoperatively. Although the transplanted tissue was functional in four patients with Graves' disease, only one became euthyroid, while the other three required supplemental hormone therapy for postoperative hypothyroidism. These findings demonstrate the ability of autotransplanted thyroid tissue to survive, function, and grow in muscle.


Author Correction: Leveraging excited-state coherence for synthetic control of ultrafast dynamics.
An amendment to this paper has been published and can be accessed via a link at the top of the paper.


The influence of duplex scanning on early patency rates of in situ bypass to the tibial vessels.
Duplex scanning was used in the follow up of 49 in situ infrapopliteal bypasses per formed for limb salvage using the valve incision method. In 19 cases (39%) the peroneal artery was the best available vessel for distal insertion; the anterior tibial and dorsalis pedis were used in 17 cases (35%) and the posterior tibial artery and tibioperoneal trunk in 13 cases (26%). In the early postoperative period three AV fistulas presented with painful skin lesions. These were located using duplex scanning and following ligation rapid resolution followed. Primary patency at one month was 87% and was increased to 96% by successful correction of four out of six failed bypasses. A total of 11 stenoses were found in five bypasses between three and 15 months postoperatively. In four of these patients the ankle/arm index decreased by at least 0.15 between examinations. A comparison was made between the spectral analysis on duplex scanning and the findings on arteriography and at operation. Peak systolic frequencies greater than 6,000 Hz were associated with 50% or greater reduction in vein bypass diameter. The majority were corrected using vein patch angioplasty. Primary cumulative patency at two years was maintained at 68% while correction of two out of seven bypass failures raised this to 80%. Duplex scanning allows precise definition of vein bypass pathology, facilitates operative correction and increases long-term limb salvage.