Gene-centric analysis identifies variants associated with interleukin-6 levels and shared pathways with other inflammation markers.
BACKGROUND- Inflammatory cytokine interleukin-6 (IL-6), a possible risk factor for coronary heart disease, has an estimated heritability of >60%, but to date few genetic variants influencing IL-6 levels are known. METHODS AND RESULTS- We used the ITMAT-Broad-Care (IBC) HumanCVD disease BeadChip in the Whitehall II study (N=4911) and British Women's Heart and Health Study (N=3445) to identify single-nucleotide polymorphisms associated with circulating IL-6 levels. Twenty-two single-nucleotide polymorphisms from 7 loci (IL6R/TDRD10, HLA-DRB1, BUD13, SEZ6L, IL1RN, TRIB3, and ABO) were associated with IL-6 (P<10(-5)), although none were associated with the IL6 gene itself. With the exception of TRIB3, all loci have been previously reported in genome-wide association studies for autoimmune and cardiovascular diseases. Fourteen single-nucleotide polymorphisms in the IL6R region in high-linkage disequilibrium (r(2)>0.9) with a nonsynonymous variant, rs2228145, were also associated with IL-6 and C-reactive protein concentration (P<10(-5)). An IL-6-specific weighted allele score explained 2% of the variance of log IL-6 levels (P=2.4410(-22)) in Whitehall II and 1% (P=1.910(-8)) in British Women's Heart and Health Studies. CONCLUSIONS- Multiple common genetic variants of modest effect influence IL-6 concentration. Several loci contain single-nucleotide polymorphisms, exhibiting overlapping associations with autoimmune and cardiovascular disorders and other circulating biomarkers. Genetic variants associated with IL-6 provide important tools for probing the causal relevance of IL-6 signaling in a range of cardiometabolic diseases.


Effects of testosterone on alpha2A-adrenergic receptor expression in the rat brain.
Androgens are involved in regulation of behaviour through intracellular mechanisms owing to their receptors. Involvement of intercellular messengers such as brain norepinephrine and adrenergic receptors (ARs) is seemed to be necessary to realise hormone-dependent behavioural effects. Castration of adult male rats, which decreases copulatory activity in the animals, was accompanied by a significant increase in 3H-clonidine (alpha2-AR agonist) binding site density in the frontal cortex. The levels of mRNA for the alpha2A-ARs (measured by RT-PCR) were increased in the brainstem of castrated males in parallel to the changes in cortical ARs densities. Testosterone treatment, that activates copulatory behaviour in castrates, down regulated alpha2A-AR mRNA levels in the brainstem and 3H-clonidine binding sites densities in the cortex, where terminals of the brain stem neurones are situated. Unlike in the brainstem, castration caused a decrease in alpha2A-AR mRNA in the cortex and testosterone up-regulated this mRNA in the cortical region. The data suggested that down-regulation of alpha2-ARs densities in the cortex that is induced by testosterone can be preferentially related to alpha2-ARs subpopulation which is expressed by the brainstem neurones and imported into the cortex by axons of these neurones.


A common human missense mutation of vesicle coat protein SEC23B leads to growth restriction and chronic pancreatitis in mice.
SEC23B is one of two vertebrate paralogs of SEC23, a key component of the coat protein complex II vesicles. Complete deficiency of SEC23B in mice leads to perinatal death caused by massive degeneration of professional secretory tissues. However, functions of SEC23B in postnatal mice and outside professional secretory tissues are unclear. In this study, we generated a Sec23b KO mouse and a knockin (KI) mouse with the E109K mutation, the most common human mutation in congenital dyserythropoietic anemia type II patients. We found that E109K mutation led to decreases in SEC23B levels and protein mislocalization. However, Sec23bki/ki mice showed no obvious abnormalities. Sec23b hemizygosity (Sec23bki/ko) was partially lethal, with only half of expected hemizygous mice surviving past weaning. Surviving Sec23bki/ko mice exhibited exocrine insufficiency, increased endoplasmic reticulum stress and apoptosis in the pancreas, and phenotypes consistent with chronic pancreatitis. Sec23bki/ko mice had mild to moderate anemia without other typical congenital dyserythropoietic anemia type II features, likely resulting from exocrine insufficiency. Moreover, Sec23bki/ko mice exhibited severe growth restriction accompanied by growth hormone (GH) insensitivity, reminiscent of Laron syndrome. Growth restriction is not associated with hepatocyte-specific Sec23b deletion, suggesting a nonliver origin of this phenotype. We propose that inflammation associated with chronic pancreatic deficiency may explain GH insensitivity in Sec23bki/ko mice. Our results reveal a genotype-phenotype correlation in SEC23B deficiency and indicate that pancreatic acinar is most sensitive to SEC23B deficiency in adult mice. The Sec23bki/ko mice provide a novel model of chronic pancreatitis and growth retardation with GH insensitivity.


Ex vivo gene transfer to mature skeletal muscle by using adenovirus helper cells.
Adenoviral gene transfer to adult skeletal muscle is hindered by several major limitations, including host immune responses and maturation-dependent loss of myofiber infectivity. Ex vivo gene delivery is more efficient than direct viral injection in surmounting maturation-dependent adenoviral transduction. Here we investigated the use of helper cells to improve the efficiency of ex vivo gene transfer to adult mouse skeletal muscle. New producer cells carrying the E1 gene of adenovirus type 5 (E32 cells) were developed using primary myoblasts from mdx mice. The E32 cells and 293 cells were infected with an E1-deleted first-generation adenovirus carrying the LacZ gene. These transduced helper cells were injected into the skeletal muscle of adult mdx and SCID mice. LacZ-positive mature myofibers were detected in the skeletal muscle of adult mice sacrificed 5 days post-injection. The gene transfer efficiency using 293 cells and E32 cells was 6.2 and 3.6 times higher than myoblast-mediated gene transfer, respectively. Ex vivo gene transfer of these cell types led to a better outcome than did direct adenoviral injection. We achieved more efficient adenoviral gene transduction by using 293 and E32 helper cells than by myoblast-mediated gene transfer and direct viral injection. These helper cells also enabled adenoviral gene transfer to mature myofibers. The mechanisms by which this method facilitated adenoviral gene transfer to mature myofibers remains unclear; however, we hypothesize that the in vivo occurrence of cytopathic effects (CPE) in the transduced 293 and E32 helper cell populations facilitated the improved adenoviral transduction of myofibers.


Kinetics and mechanism of 17β-estradiol chlorination in a pilot-scale water distribution systems.
The kinetics and mechanisms of 17β-estradiol (E2) chlorination in water distribution systems (WDS) were studied. We examined the impacts of different factors, including pH, temperature, humic acid concentration and type, and flow velocity. The experimental results showed that the rate constants in beaker tests and WDS were described by a pseudo-first-order model. pH had the greatest impact on E2 chlorination in the beaker tests. However, temperature had the greatest impact on E2 chlorination in WDS. Mechanistic analysis of E2 chlorination showed that chlorine attacked E2 in three stages: 1) halogenation of the aromatic ring, 2) cleavage of the benzene moiety and chlorine or bromine substitution formation, and 3) formation of trihalomethanes (THMs) and halogenated acetic acids (HAAs) from phenolic intermediates through benzene ring opening with chlorine and/or bromine substitution of hydrogen on the carbon atoms. In the third stage, the concentrations of THMs and HAAs increased rapidly.


Early lymphocyte recovery at 28 d post-transplant is predictive of reduced risk of relapse in patients with acute myeloid leukemia transplanted with peripheral blood stem cell grafts.
Allogeneic hematopoietic cell transplantation (HCT) is potentially curative for acute myeloid leukemia (AML). Impact of lymphocyte recovery on post-transplant outcomes has been suggested but reports are conflicting. We evaluated the impact of lymphocyte recovery at 28 d post-HCT in 191 AML patients using peripheral blood stem cells as graft. Patients were divided into those with absolute lymphocyte count (ALC) ≥ 0.5 × 10(9) /L (n = 111, 58%; high ALC group) and those with ALC < 0.5 × 10(9) /L (n = 80, 42%; low ALC group), at day 28 post-transplant. With a median follow-up of 49 months, overall survival (OS) was significantly improved in the high ALC group (59% at 3 yr) vs. patients with low ALC (40% at 3 yr, P = 0.03). Cumulative incidence of relapse (CIR) was significantly lower in the high ALC group (16% at 3 yr) vs. low ALC group (36% at 3 yr, P = 0.001). Multivariable analysis for CIR demonstrated high ALC group as an independent factor decreasing relapse risk (P = 0.03, HR = 0.49, 95% CI = 0.26-0.92). Multivariable analysis for OS and non-relapse mortality did not demonstrate ALC ≥ 0.5 × 10(9) /L at 28 d post-transplant to be predictive. We conclude that lymphocyte recovery with ALC ≥ 0.5 × 10(9) /L at day 28 post-transplant is associated with less relapse in AML patients undergoing allogeneic peripheral blood HCT, but without survival benefit.


The type of gastrectomy and modified frailty index as useful predictive indicators for 1-year readmission due to nutritional difficulty in patients who undergo gastrectomy for gastric cancer.
Patients who undergo gastrectomy for gastric cancer (GC) are likely to have nutritional difficulty after surgery. Readmission due to nutritional difficulty is common in such patients. Thus, in this study, we aim to identify the predictive indicators for readmission due to nutritional difficulty in patients who underwent gastrectomy for GC. We retrospectively reviewed surgical outcomes in 516 consecutive patients who underwent gastrectomy for GC. The readmission rate within 1 year was 13.8%. Readmission due to nutritional difficulty was observed in 20 patients (3.9%); it was determined as the second leading cause of readmission. Multivariate analysis revealed that the type of gastrectomy and the modified frailty index (mFI) were independent predictive indicators of readmission due to nutritional difficulty. Patients were assigned 1 point for each predictive indicator, and the total points were calculated (point 0, point 1, or point 2). The readmission rates due to nutritional difficulty were 1.2%, 4.7%, and 11.5% in patients with 0, 1, and 2 points, respectively (P = 0.0008). The readmission rate due to nutritional difficulty was noted to be high in patients who underwent total or proximal partial gastrectomy with high mFI. Intensive follow-up and nutritional support are needed to reduce readmissions due to nutritional difficulty. Reduced readmission rates can improve patient quality of life and reduce medical costs.


Validation of an efficient and continuous urodynamic monitoring system for awake, unrestrained, chronic rodent studies.
The postvoid residual (PVR) is an important measure of bladder function, but obtaining PVR is burdensome because bladder volume must be measured at the time of voiding. The PVR measurement problem has led to experimental tricks in animal studies (infusing the bladder at supraphysiological rates and limiting animal observation windows) to keep the number of observed voids statistically robust while reducing the time an experimenter must be present. Our solution to the PVR measurement problem is a system called Automatic Monitoring for Efficient, Awake, Sensitive, Urine Residual Estimation (AMEASURE). AMEASURE combines metabolic cages and optimization algorithms to estimate continuously PVR for every voiding event 24 h/day for multiple weeks, without artificial bladder infusion, continuous experimenter supervision, anesthesia, or restraints. Using AMEASURE, we obtained voided volumes, PVRs, and other urodynamic parameters continuously for 21 days in 10 healthy female Sprague-Dawley rats. Importantly, this required only one manual measurement of animals' bladder volume every 12 h. We validated the accuracy of the system experimentally and in simulation. We detected marked differences in voiding frequency and efficiency between light and dark cycles and found that voiding frequency increased over time during the dark cycle (but not the light cycle), due to surgical recovery, cage acclimation, and socialization. This tool enhances the relevance of rodent models to the study of human lower urinary tract by expanding observation periods and obviating the need to infuse the bladder and facilitates the study of conditions for which behavioral, social, or circadian factors play essential roles.


[Nondetectable prostate cancer in radical prostatectomy specimens].
Since the widespread use of prostate-specific antigen, early diagnosis of prostate cancer at an early stage has been increased. Consequently, a greater frequency of low-volume disease or no tumor has been found in radical prostatectomy specimen. In the present study authors analyzed patients classified as pT0 after radical prostatectomy at their center. 1134 radical prostatectomies were evaluated retrospectively at the author's center between 1996 and 2010. If there was no evidence of prostate cancer in the specimen, patient was staged as pT0. Patients were divided into two groups: received neoadjuvant hormonal therapy or not. Overall 32 (2.8%) patients were staged as pT0. The rate of pT0 staging was 9.3% and 1.2% in the hormonally treated group and non-hormonally treated group. False-positive prostate biopsy was found in 2 cases. The rate of pT0 staging was higher in patients with incidental prostate cancer, low Gleason score and enlarged prostate. Biochemical relapse was observed in 3 pT0 patients in the hormonally treated group, among them there was one clinical relapse. In non-hormonally treated group no recurrence was detected. The rate of pT0 staging was higher in the hormonally treated group. Because of biochemical and clinical relapse despite vanishing prostate cancer phenomenon, these cases are considered not to be true pT0. On the basis of present study and other reports the rate of pT0 staging is about one percentage in non-hormonally treated patients. Prognosis of these patients is excellent.


Corrigendum to "A comprehensive review on modelling the adsorption process for heavy metal removal from waste water using artificial neural network technique" [Heliyon 9(4) (April 2023) e15455].
[This corrects the article DOI: 10.1016/j.heliyon.2023.e15455.].


Immunogenicity Characterization of the Recombinant gI Protein Fragment from Pseudorabies Virus and an Evaluation of Its Diagnostic Use in Pigs.
Serological testing is an important method for the diagnosis of pseudorabies virus (PRV) infection. We aimed to investigate the envelope glycoprotein I (gI) of PRV, a strong immunogen, and its potential as an efficient and low-cost diagnostic reagent. In this study, the DNA of the PRV SC strain was used as the template, and the recombinant fragment of gI (633 bp) was amplified via PCR using synthetic primers, and was then ligated into the pET-30a expression vector. The constructs were transferred into Escherichia coli (E. coli) for prokaryotic expression, and the antigenicity of the expression products was identified by Western blot analysis with pig positive serum against PRV. The recombinant protein was purified by a Ni column, and BALB/c mice were immunized with purified gI protein to obtain anti-gI-positive serum. After PK-15 cells had been infected by PRV for 48 h, the immunogenicity of purified gI protein was identified with a fluorescence immunoassay using anti-gI mouse serum. The recombinant plasmid (pET-30a-gI) was expressed, and the native gI protein was obtained after denaturation by urea and renaturation by dialysis. A small-scale ELISA test containing 1.0 µg/mL of purified gI protein was designed to evaluate pig serum (80 samples), and the results of the ELISA test were compared to those of competitive ELISA (cELISA) tests using IDEXX Kits, which resulted in 97.5% consistency. The results suggested that the truncated gI protein may be a potential diagnostic reagent.


Regenerative Medicine in COVID-19 Treatment: Real Opportunities and Range of Promises.
Novel coronavirus disease (COVID-19) has attracted much attention around the world due to its rapid transmission among humans and relatively high mortality rate. Studies are increasing to find the best therapeutic approach for the disease and its management. Regenerative medicine offers various cell-tissue therapeutics and related products, such as stem cell therapy, natural killer (NK) cell therapy, Chimeric antigen receptor (CAR) T cell therapy, exosomes, and tissue products. Interestingly, mesenchymal stem cells (MSCs) can reduce inflammatory symptoms and protect against cytokine storm, which critically contributes to the COVID-19 progression. Notably, having the potentials to exert cytotoxic effects on infected cells and induce interferon production probably make NK cells a candidate for COVID-19 cell therapy. Besides, exosomes are one of the crucial products of cells that can exert therapeutic effects through the induction of immune responses and neutralizing antibody titers. The paper aims to briefly consider current options for COVID-19 therapy to show that there is no specific cure for COVID-19, and then assess the real opportunities and range of promises regenerative medicine can provide for specific treatment of COVID-19. Graphical Abstract Therapeutic Potential of Regenerative Medicine against COVID19.


Cell kinetics of the malignant evolution of meningothelial meningioma.
A meningotheliomatous meningioma occurred in the right parietal parasagittal region of a 43-year-old woman. A total excision was carried out. Three years and 9 months later, one parasagittal and two falx tumors in close proximity recurred and were excised. The falx tumors showed the features of an anaplastic transitional meningioma with increased mitoses. The falx meningioma recurred locally and repeatedly, despite radiation therapy and chemoimmunotherapy. She died 7 years and 8 months after the onset. At autopsy, the tumor was found to have metastasized via the cerebrospinal fluid to the spinal cord and pituitary stalk. Extracranial metastases were not evident. At the fourth craniotomy, bromodeoxyuridine (BrdU) was administered intravenously to label tumor cells in the S phase of the cell cycle. The BrdU-labeled cells accounted for 9.0%; they are usually less than 1% in benign meningiomas. Meningothelial meningiomas with a high labeling index recur rapidly, even after Simpson's grade 2 removal, and treatment for a malignant meningioma should be instituted.


A histochemical study of the distribution of aldehyde dehydrogenase activity in brain structures of rats with genetically different alcohol-related behaviour.
Brain samples from rats genetically selected for high or low voluntary alcohol intake (AA and ANA strains) or for differences in alcohol-induced motor incoordination (AT and ANT strains) were analyzed by histochemistry for aldehyde dehydrogenase (EC 22.214.171.124; ALDH) activity in various CNS structures. All strains exhibited the highest ALDH activities in neurons of the mesencephalic tract of trigeminal nerve nucleus and in spinal cord motoneurons, while the lowest activities were observed in the somatosensory cortex. Although the general distribution pattern of ALDH activity was similar in the genetically selected strains, some potentially important differences were observed. AA rats with high voluntary alcohol consumption had lower ALDH activity (with acetaldehyde as substrate) in the neuropil of the olfactory tubercle but higher activity (with benzaldehyde as substrate) in the spinal cord motoneurons, Purkinje cells and capillary endothelium of the cerebellum as compared to the corresponding structures from the alcohol avoiding ANA rats. Alcohol-resistant AT rats had higher ALDH activity, with benzaldehyde, in most CNS structures than did the alcohol-sensitive ANT's, significantly so in the lamina II of the somatosensory cortex and the neurons of the lateral hypothalamic area. This relationship was also found with acetaldehyde as substrate in the neurons of the hypothalamic arcuate nuclei and in cerebellar capillaries, but the ANT's had the higher activity in the neurons of the cerebral cortex V lamina. We suggest that some of the differences observed may relate to the differences between the rat strains with respect to voluntary alcohol intake and alcohol-induced motor incoordination.


Rapid Macular Thinning Is an Early Indicator of Hydroxychloroquine Retinal Toxicity.
To demonstrate rapid macular thinning as an early and objective sign of hydroxychloroquine retinopathy. Retrospective case cohort. Cohort of 301 patients receiving long-term hydroxychloroquine therapy at Kaiser Permanente Northern California who underwent a minimum of 4 OCT studies that included Early Treatment Diabetic Retinopathy Study (ETDRS) retinal thickness values over a minimum of 4 years. Creation of sequential retinal thickness plots to show the rate of change in macular thickness within ETDRS regions. Identification of rapid macular thinning, comparison of patients with rapid thinning to those with stable macular thickness, and comparison of rapid thinning patients with and without conventional OCT or 10-2 visual field signs of hydroxychloroquine toxicity. Retina thinning in 219 stable patients receiving long-term hydroxychloroquine therapy averaged (mean ± standard deviation) 0.62 ± 0.45 μm/year, whereas 82 patients showed a period of relatively linear rapid thinning with a loss of 3.75 ± 1.34 μm/year. Of the patients with rapid thinning, 38 eventually demonstrated conventional OCT or 10-2 visual field signs of hydroxychloroquine retinal toxicity. The cumulative retinal thinning in these patients was 25.1 ± 6.2 μm compared with 15.7 ± 4.0 μm in those without conventional toxicity (P < 0.01). Retinal thickness remains stable for many years in most patients receiving long-term hydroxychloroquine therapy, but after a critical point, the retina may begin to thin rapidly. Sequential plots of inner and outer ETDRS ring macular thickness provide objective evidence of this early structural change several years before conventional signs appear. This approach can alert patients and prescribing physicians to potential retinal damage and uses readily available OCT measurements that could be automated by manufacturers for use in comprehensive eye care settings.


The convergent lady beetle, Hippodamia convergens Guérin-Méneville and its endoparasitoid Dinocampus coccinellae (Schrank): the effect of a microsporidium on parasitoid development and host preference.
Convergent lady beetles, Hippodamia convergens Guérin-Méneville are host to the braconid endoparasitoid, Dinocampus coccinellae (Schrank) and the microsporidian pathogen, Tubulinosema hippodamiae. The interrelationship between the endoparasitoid and the pathogen in H. convergens adults under laboratory conditions was examined by quantifying the effect of microsporidiosis on D. coccinellae development and host preference. Uninfected wasps were provided either uninfected or T. hippodamiae-infected beetles as hosts and the development of their progeny was observed over 30 days. The duration of endoparasitoid development from egg deposition in the host until adult eclosion for D. coccinellae did not differ significantly, regardless of the infection status of the host beetle. All wasp progeny that developed within, and emerged from, T. hippodamiae-infected beetles were infected with the microsporidian pathogen (n = 48; 100% transmission). Infected D. coccinellae adults were also provided either uninfected or T. hippodamiae infected host beetles so that the development of their progeny could be assessed over 30 days. Endoparasitoid development did not differ significantly; however, a significantly greater proportion of beetles stung by microsporidia-infected wasps did not contain an endoparasitoid larva when dissected at the end of the 30-day trial when compared to those stung by uninfected wasps. This suggests that the pathogen may reduce wasp fecundity or egg viability. Examination of paraffin-embedded D. coccinellae adult tissues revealed an extensive microsporidian infection throughout all major organs and tissues with exception of the ovary. During host choice trials, uninfected and microsporidia-infected D. coccinellae adults pursued, took an ovipositional stance, and attacked uninfected beetles more often than microsporidia-infected hosts but these observations did not differ significantly (P > 0.05).


Emergency department visits in older patients: a population-based survey.
Given the higher incidence of emergency conditions in older inhabitants, the global increase in aged population will pose a challenge for emergency services. In this study we examined the burden caused to emergency health care by the aged population. Consecutive patients aged 80 years or over visiting a high-volume, collaborative emergency department (ED) between 2015 and 2016 were included. The key factors under analysis were the incidence of emergency conditions and costs associated with emergency care. A total of 6944 patients (median age 85 years, range 80-104 years; 67% female) aged ≥80 years representing 1.5% of the local population, made 17,769 ED visits during the two-year observation period accounting for 15% of all ED visits. Forty-two percent (n = 2884) of patients had a single ED visit, whereas 8.2% (n = 570) made ≥5 ED visits/year for a total of 1400 visits (7.9%). Thirty-two percent of those aged ≥80 years required ED services each year. The number of ED visits increased with age (p < 0.001); and was 768/1000 person-years among octogenarians and 1007/1000 among nonagenarians, in comparison to 233/1000 among those aged < 80 years. One in five of the study population were discharged with non-specific diagnoses. Typical diagnoses included pneumonia (4.8%), malaise and fatigue (4.5%) and heart failure (4.3%). Non-specific diagnoses were frequent, and examination of patients with non-specific diagnoses incurred costs similar to or higher than those of other patients. The mean cost per ED visit in older patients was 422 €. We demonstrated a high incidence of emergency department visits in older patients. While our aim was not to solve how the growing demand should be met, it seems unlikely that increasing ED resources is feasible. Instead, the focus should be on chronic care of the aged and prevention of potentially avoidable ED visits.


Progressing Sclerosing Mesenteritis (Mesenteric Panniculitis) Mimics Progression of Malignancy After Neoadjuvant Chemotherapy for Gastric Adenocarcinoma on Serial 18F-FDG PET/CT.
A 62-year-old man was diagnosed with a moderately differentiated gastric adenocarcinoma in the proximal stomach. A staging 18F-FDG PET/CT showed an intensely FDG-avid gastric mass, as well as a mildly FDG-avid misty nodular mesentery. After 3 cycles of neoadjuvant chemotherapy, a follow-up PET/CT showed partial response of the gastric primary, with increase in the size of nodules in the mesentery and increased FDG uptake, raising concern of secondary malignancy. Biopsy of the mesentery revealed xanthogranulomatous inflammation, consistent with sclerosing mesenteritis.


Intraorbital optic nerve signal hyperintensity on magnetic resonance imaging sequences in perioperative hypotensive ischemic optic neuropathy.
A 61-year-old man experienced severe bilateral posterior ischemic optic neuropathy after cardiac bypass surgery. Routine magnetic resonance imaging sequences were normal, but diffusion-weighted and fluid-attenuated inversion recovery (FLAIR) sequences showed abnormal hyperintensity within both intra-orbital optic nerves. This imaging abnormality has not been previously reported in this setting.


Correlating state-specific and national trends in breast reconstruction after Medicaid expansion: A decade-long update on the Affordable Care Act's impact.
While disparities in access to reconstruction persist, a comprehensive analysis comparing state-based outcomes and national patterns in breast reconstruction as a result of Medicaid expansion has never been examined. In this study, we investigated how breast reconstruction rates changed as a result of Medicaid expansion and compared these state-based findings to national counterparts. Patient data from the Healthcare Cost and Utilization Project among states that chose to expand Medicaid were compared with those from states that did not expand. The difference-in-differences estimate of expansion to nonexpansion states was 7.05 (p = 0.10) for implant-based reconstruction, -11.56 (p = 0.01) for autologous reconstruction, and -7.08 (p = 0.18) for overall reconstruction. Comparing rates of nonexpansion states to national trends yielded estimates of -0.06 (p = 0.04), 0.06 (p = 0.01), and 0.004 (p = 0.90) for implant-based, autologous, and overall breast reconstruction, respectively. Similarly, comparing rates of expansion states to national trends yielded estimates of 0.02 (p = 0.38), -0.05 (p = 0.03), and -0.02 (p = 0.44) for implant-based, autologous, and overall breast reconstruction, respectively. In this study on national health policy, Medicaid expansion was associated with a significant increase in autologous rates while state-specific trends alone did not appear to predict the national outcomes of sweeping legislative changes that were differentially applied among states.


Three-dimensional multifractal analysis of trabecular bone under clinical computed tomography.
An adequate understanding of bone structural properties is critical for predicting fragility conditions caused by diseases such as osteoporosis, and in gauging the success of fracture prevention treatments. In this work we aim to develop multiresolution image analysis techniques to extrapolate high-resolution images predictive power to images taken in clinical conditions. We performed multifractal analysis (MFA) on a set of 17 ex vivo human vertebrae clinical CT scans. The vertebrae failure loads (FFailure) were experimentally measured. We combined bone mineral density (BMD) with different multifractal dimensions, and BMD with multiresolution statistics (e.g., skewness, kurtosis) of MFA curves, to obtain linear models to predict FFailure. Furthermore we obtained short- and long-term precisions from simulated in vivo scans, using a clinical CT scanner. Ground-truth data - high-resolution images - were obtained with a High-Resolution Peripheral Quantitative Computed Tomography (HRpQCT) scanner. At the same level of detail, BMD combined with traditional multifractal descriptors (Lipschitz-Hölder exponents), and BMD with monofractal features showed similar prediction powers in predicting FFailure (87%, adj. R2 ). However, at different levels of details, the prediction power of BMD with multifractal features raises to 92% (adj. R2) of FFailure. Our main finding is that a simpler but slightly less accurate model, combining BMD and the skewness of the resulting multifractal curves, predicts 90% (adj. R2) of FFailure. Compared to monofractal and standard bone measures, multifractal analysis captured key insights in the conditions leading to FFailure. Instead of raw multifractal descriptors, the statistics of multifractal curves can be used in several other contexts, facilitating further research.


Some remarks on assessing morphologic changes in old, demented people.
Some aspects of quantitative assessment of senile and vascular changes in brains of old, demented people are discussed. Particular emphasis is placed upon neuritic plaque count and upon the amount of softened brain parenchyma relative to the degree of intellectual deterioration. The functional significance of the topography of ischemic lesions is stressed in some cases.


Update on the Quality and Health Literacy Demand of Diet-Related Videos on YouTube for People With Polycystic Kidney Disease.
To describe the quality and health literacy demand of publicly available diet-related videos on YouTube for people with polycystic kidney disease (PKD). This desk-based content analyzed involved a comprehensive search of YouTube using key words relating to the dietary management of people with PKD. The health literacy demand was evaluated using the Patient Education Material Assessment Tool (PEMAT) for audio-visual materials. A PEMAT score of >70% is indicative of videos that are understandable and actionable. A total of 15 videos were evaluated (median 1.28 minutes duration (range: 55 seconds-81 minutes). The majority were produced by reputable sources (such as a PKD-related charity (n = 12) or a nephrologist (n = 2). The quality of the videos was high with the majority (14/15) providing evidence-based advice. The understandability and actionability of the videos were considered low (median PEMAT score 67% (interquartile range: 51.4-81.7) and 66.83% (interquartile range: 50-67), respectively. There has been a small increase in the number of diet-related videos for people with PKD in the past 5 years. The majority provide evidence-based information, which is also a substantial improvement since the last analysis in 2015. Unfortunately, the health literacy demand of these resources remains suboptimal, and future work should be directed to improving the understandability and actionability of videos.


Use of confocal scanning laser microscopy to measure the concentrations of aerial and penetrative hyphae during growth of Rhizopus oligosporus on a solid surface.
In order to develop a method for use in investigations of spatial biomass distribution in solid-state fermentation systems, confocal scanning laser microscopy was used to determine the concentrations of aerial and penetrative biomass against height and depth above and below the substrate surface, during growth of Rhizopus oligosporus on potato dextrose agar. Penetrative hyphae had penetrated to a depth of 0.445 cm by 64 h and showed rhizoid morphology, in which the maximum biomass concentration, of 4.45 mg dry wt cm(-3), occurred at a depth of 0.075 cm. For aerial biomass the maximum density of 39.54 mg dry wt (-3) occurred at the substrate surface. For both aerial and penetrative biomass, there were two distinct regions in which the biomass concentration decayed exponentially with distance from the surface. For aerial biomass, the first exponential decay region was up to 0.1 cm height. The second region above the height of 0.1 cm corresponded to that in which sporangiophores dominated. This work lays the foundation for deeper studies into what controls the growth of fungal hyphae above and below the surfaces of solid substrates.


An accurate and efficient method for occlusal tooth wear assessment using 3D digital dental models.
Tooth or material wear in a dentition is a common finding that requires timely diagnosis for management and prevention of further loss or associated esthetic or functional impairment. Various qualitative and quantitative methods have been suggested to measure tooth or material wear, but they present with limitations, such as imprecision, subjectivity, or high complexity. Here we developed and assessed an efficient 3D superimposition method to accurately measure occlusal tooth wear on 3D digital dental models. For this purpose, teeth on plaster casts were manually grinded on their occlusal surfaces to simulate various degrees of tooth wear. The casts were scanned using a surface scanner. Grinded tooth crowns (T1) were segmented and compared to the original crowns (T0) using five 3D surface superimposition techniques and a gold standard technique (GS). GS measurements were obtained by using intact adjacent structures as superimposition references. The technique of choice (complete crown with 30% estimated overlap of meshes) showed the best reproducibility (maximum difference < 0.050 mm3) and excellent agreement with the GS technique (median difference: 0.032 mm3). The suggested 3D superimposition method offers a highly efficient and accurate tool for tooth wear assessment, which could be applicable to clinical conditions.


Addressing the psychosocial wellbeing of teenage children of cancer patients and survivors.
Thomas is 13 years old. His parents report a sharp decline in his school grades caused, according to his teachers' opinions, by listlessness and lack of concentration. The parents of Julia, 16 years old, describe her as restless, evasive, isolated, and withdrawn from others and from her usual activities. Linda, 18 years old, is described by her parents as indecisive, uncertain, and almost lethargic. Normally resolute and a high academic achiever, she appears locked in herself, unable to make choices. We first learned about them through the accounts of their concerned parents. Claire, 19 years old, lost weight and exercised hard enough to induce amenorrhea after her young mother underwent treatment for breast cancer, including antihormonal treatment. These four teenagers have in common a parent diagnosed with cancer, undergoing or having just completed treatment.


Brain tumor survival: results from the National Cancer Data Base.
Hospital-based data reported to the National Cancer Data Base (NCDB) were available for over 60,000 patients with a primary brain tumor diagnosed from 1985-1988 and 1990-1992. The most common histologies were glioblastomas, astrocytomas and meningiomas. Five-year survival rates for these tumors were 2%, 30% and 70% respectively. Histology, age at diagnosis, behavior, and location were important variables in estimating survival. Comparisons with population-based registry data suggest that the malignant tumors are well represented in NCDB, but the benign histologies are under-reported. Survival estimates for the malignant tumors are comparable to previously reported studies. The NCDB provides recent information on brain tumor distribution and survival patterns not available in other large databases.


[Making a diagnosis of different lesions of vocal fold mucosa by contact endoscopy--the first usage in our clinical practice].
During laryngomicroscopy, the superficial layers of vocal fold epithelium can be examined in vivo and in situ by contact endoscopy. Methylene blue is applied initially to stain the epihelial cells of the vocal folds. When in contact with mucosal tissue, this endoscope provides 60 and 150 times magnification and clear visualization of cellular patterns of the superficial epithelial layers. For the first time in our laryngological clinical practice, we confirmed a number of previosly established parametars such as regularity and arrangement of the epithelium, nucleus contour, and nucleus-cytoplasm ratio, what all enable recognition and easy evaluation of different clinical conditions such as chronic laryngitis, Reinke's edema, papiloma dysplasia or vocal fold carcinoma. The advantage of contact endoscopy in vivo and in situ allows for detailed scan and mapping of all cell changes of the whole mucosa surface. All these features definitely classify the contact endoscopy into additional diagnostic methods in laryngology.


Abnormal sharp transients on electroencephalograms in preterm infants with periventricular leukomalacia.
To determine the clinical significance of abnormal sharp transients other than positive rolandic sharp waves (PRS), electroencephalograms were used for the diagnosis of periventricular leukomalacia (PVL). We evaluated 126 electroencephalograms from 93 preterm infants; 31 infants had PVL, and 62 were control infants. Frontal sharp waves (FS) were defined as sharp transients of positive polarity with an amplitude >100 microV. Occipital sharp waves (OS) were defined as those of negative polarity with an amplitude >150 microV. FS, OS, or PRS were considered to be present when there were >0.1 per minute. The number of FS per minute was significantly higher in the PVL group than in the control group during days 0 to 4 and 5 to 7. The number of OS per minute was also significantly higher in the PVL group than in the control group during days 0 to 4, 5 to 7, and 8 to 14. The sensitivity of FS or OS was relatively high but that of PRS was low. The presence of two or more types of abnormal sharp transients was correlated with a poor outcome. FS or OS may be useful for predicting which infant will have PVL.


Depletions of central norepinephrine by intraventricular xylamine in rats.
A series of neurochemical studies evaluated the selectivity of the depletions of norepinephrine produced by intraventricular injections of xylamine. Brain monoamines were assayed by liquid chromatography with electrochemical detection. With ether anesthesia, bilateral injections of 50 or 100 micrograms xylamine reduced hippocampal levels of norepinephrine and serotonin without affecting striatal dopamine or serotonin. The depletion of hippocampal norepinephrine was significant as soon as 24 h. Animals treated with the combination of 20 mg/kg fluoxetine and 100 micrograms xylamine had selective depletions of central norepinephrine with no significant changes in serotonin.


Development of a decentralized cohort for studying post-acute sequelae of COVID-19 in India in the Data4life Study.
Decentralized, digital health studies can provide real-world evidence of the lasting effects of COVID-19 on physical, socioeconomic, psychological, and social determinant factors of health in India. Existing research cohorts, however, are small and were not designed for longitudinal collection of comprehensive data from India's diverse population. Data4Life is a nationwide, digitally enabled, health research initiative to examine the post-acute sequelae of COVID-19 across individuals, communities, and regions. Data4Life seeks to build an ethnically and geographically diverse population of at least 100,000 participants in India. Here we discuss the feasibility of developing a completely decentralized COVID-19 cohort in India through qualitative analysis of data collection procedures, participant characteristics, participant perspectives on recruitment and reported study motivation. As of June 13th, 2022, more than 6,000 participants from 17 Indian states completed baseline surveys. Friend and family referral were identified as the most common recruitment method (64.8%) across all demographic groups. Helping family and friends was the primary reason reported for joining the study (61.5%). Preliminary findings support the use of digital technology for rapid enrollment and data collection to develop large health research cohorts in India. This demonstrates the potential for expansion of digitally enabled health research in India. These findings also outline the value of person-to-person recruitment strategies when conducting digital health research in modern-day India. Qualitative analysis reveals opportunities to increase diversity and retention in real time. It also informs strategies for improving participant experiences in the current Data4Life initiative and future studies.


A methodology to model causal relationships on offshore safety assessment focusing on human and organizational factors.
Focusing on people and organizations, this paper aims to contribute to offshore safety assessment by proposing a methodology to model causal relationships. The methodology is proposed in a general sense that it will be capable of accommodating modeling of multiple risk factors considered in offshore operations and will have the ability to deal with different types of data that may come from different resources. Reason's "Swiss cheese" model is used to form a generic offshore safety assessment framework, and Bayesian Network (BN) is tailored to fit into the framework to construct a causal relationship model. The proposed framework uses a five-level-structure model to address latent failures within the causal sequence of events. The five levels include Root causes level, Trigger events level, Incidents level, Accidents level, and Consequences level. To analyze and model a specified offshore installation safety, a BN model was established following the guideline of the proposed five-level framework. A range of events was specified, and the related prior and conditional probabilities regarding the BN model were assigned based on the inherent characteristics of each event. This paper shows that Reason's "Swiss cheese" model and BN can be jointly used in offshore safety assessment. On the one hand, the five-level conceptual model is enhanced by BNs that are capable of providing graphical demonstration of inter-relationships as well as calculating numerical values of occurrence likelihood for each failure event. Bayesian inference mechanism also makes it possible to monitor how a safety situation changes when information flow travel forwards and backwards within the networks. On the other hand, BN modeling relies heavily on experts' personal experiences and is therefore highly domain specific. "Swiss cheese" model is such a theoretic framework that it is based on solid behavioral theory and therefore can be used to provide industry with a roadmap for BN modeling and implications. A case study of the collision risk between a Floating Production, Storage and Offloading (FPSO) unit and authorized vessels caused by human and organizational factors (HOFs) during operations is used to illustrate an industrial application of the proposed methodology.


A novel voltammetric sensor for amoxicillin based on nickel-curcumin complex modified carbon paste electrode.
The electrocatalytic oxidation of amoxicillin was investigated on a nickel-based (Ni(II)-curcumin) chemically modified electrode. This modified electrode was prepared by electropolymerization of complex (curcumin = 1,7-bis[4-hydroxyl-3-methoxyphenyl]-1,6-heptadiene-3,5-dione) in alkaline solution. For the first time, the catalytic oxidation of amoxicillin was demonstrated by cyclic voltammetry, chronoamperometry, chronocoulometry and amperometry methods at the surface of this modified carbon paste electrode. The obtained results showed that NiOOH acts as an electrocatalyst for oxidation of amoxicillin. This electrocatalytic oxidation exhibited a good linear response for amoxicillin concentration over the range of 8 × 10⁻⁶-1×10⁻⁴ M with a detection limit of 5 × 10⁻⁶ M. Therefore, this electrocatalytic method was used as a simple, selective and rapid method able to determine amoxicillin in pharmaceutical preparations and biological media.


Are Palliative Care Volunteers Special People? A Comparative Observational Study Exploring Their Sociodemographic Profile and Psychological Resources.
Faced with rising needs for patient support, palliative care is shifting towards a more community-based approach. Yet the profile of volunteers in this field is poorly known. To explore psychosocial characteristics of palliative care volunteers, by comparing them to two groups, volunteers from the health and social sector without contact with palliative patients, and people from the general population without volunteer activity. Observational comparative study. Data were analyzed using descriptive statistics and logistic regression analysis. The 3 groups of participants were recruited in France between September 2020 and June 2021. They completed an online survey assessing sociodemographic characteristics and psychological resources (self-efficacy, hope, optimism, resilience, mindfulness, self-compassion, empathy, forgivingness, and gratitude). Data from 559 participants were analyzed. Palliative care volunteers had overall significantly higher levels of psychological resources than control groups. Multivariate analysis revealed, among sociodemographic and psychological characteristics that showed differences between groups at the univariate level, the most efficient predictors of group status. The best criteria to distinguish palliative care volunteers from health and social sector volunteers were older age, having received training, and lower levels of self-compassion. To distinguish palliative care volunteers from people with no volunteer activity, older age and non-active professional status were the best criteria. Palliative care volunteers displayed more psychological resources than controls. However, older age and being trained for volunteering, emerged as stronger factors for distinguishing palliative care volunteers from controls. Pursuing research about these volunteers should facilitate recruitment, training, and retention.


Lethal comorbidity with genital anomaly in the infant.
Genital anomaly (GA) carries a risk of life-threatening comorbidity which is not well defined and is under reported. We aimed to determine the associated anomalies in children with GA who died. We retrospectively reviewed the deaths among all GA patients presenting to our institution over 32 years (1970-2001). Seventy children with exstrophy/epispadias were excluded, leaving 200 patients, of whom 26 had died. Hospital and postmortem records were examined for biographical data, karyotype, morphological anomalies and causes of death. Sixty-eight out of 200 patients had congenital adrenal hyperplasia (CAH), with two deaths, from cardiomyopathy and encephalitis. Of the 132 non-CAH patients, 24 (17%) died. Sixteen deaths were in the non-CAH intersex group of patients with GA. There were eight deaths in the non-intersex genital anomaly group, which included five with a cloacal anomaly (+/- VATER association). Fifteen had major cardiac anomalies and 10 had facial dysmorphism suggesting a syndrome. Two infants died of renal failure with Denys-Drash syndrome. Infants with a non-CAH intersex GA have a high risk of mortality, particularly from an associated cardiac anomaly. This mortality may decrease in the future with cardiac surgical advances. There is a significant mortality in infants with non-intersex GA (anorectal malformations and cloacal anomalies) also due to complex cardiac and/or renal anomalies. Major chromosomal aberrations and facial dysmorphism were noted in some patients in both study groups. GA associated with testicular dysgenesis and/or imperforate anus requires screening for cardiac defects, chromosomal anomalies and dysmorphology syndromes.


The effect of mesenchymal stem cells and surgical angiogenesis on immune response and revascularization of acellular nerve allografts in a rat sciatic defect model.
Increasing evidence demonstrates an interplay between neoangiogenesis and immune cells. We investigated the immune response and revascularization of acellular nerve allografts (ANA) after combined stem cell delivery and surgical angiogenesis in a rat model. Unilateral sciatic nerve defects in 60 Lewis rats were repaired with (I) autografts, (II) ANAs, and (III) ANAs wrapped within a pedicled superficial inferior epigastric artery fascial flap to induce surgical angiogenesis, combined with seeding of either (IV) undifferentiated mesenchymal stem cells (uMSCs) or (V) MSCs differentiated into Schwann cell-like cells. Immune cell phenotyping was performed on days 7 and 14. The vascular volume of nerves was measured by microcomputed tomography at 12 and 16 weeks. On day 7, helper T cells (CD4+) were significantly increased in groups IV and V compared to group I. Regulatory T cells (CD4+CD25+) were significantly higher in groups III-IV, and cytotoxic T cells (CD8+) were significantly reduced in groups IV and V compared to group II, respectively. Group II demonstrated the highest levels of natural killer cells (CD161+) compared to groups III-V. On day 14, group IV demonstrated the highest CD4/CD8 ratio. Vascular volume was significantly higher in groups III-V compared to group II at 12 weeks and groups IV and V compared to group II at 16 weeks. The CD4/CD8 ratio demonstrated a positive correlation to vascular volumes at 12 weeks. Early favorable immune responses were observed in ANAs treated with surgical angiogenesis with or without stem cell delivery and demonstrated improved vascularity at longer follow-up.


A reliable, non-invasive technique for measuring growth in tadpoles exposed to salt.
The use of chemical de-icers raises salt levels in roadside streams and ponds, which has adverse effects on tadpole development. Experiments on the effects of de-icers on tadpole development are often hampered by difficulties measuring body size without introducing handling stress that may skew results or cause unintended mortality. We have found a linear relationship between surface area and body mass in tadpoles that is unaffected by exposure to salt. Measuring surface area is therefore a suitable technique whose use should be encouraged when investigating the effects of salt exposure on tadpole growth and development.


Do Biologic Treatments for Psoriasis Lower the Risk of Psoriatic Arthritis? A Systematic Review.
The effectiveness of biologic treatments in slowing the progression of psoriatic arthritis is well established, but there is limited and conflicting evidence on their ability to prevent the development of psoriatic arthritis in patients with psoriasis. The objective of this review was to evaluate the role of biologic treatment for psoriasis in preventing or delaying subsequent psoriatic arthritis. A literature search was performed using MEDLINE (PubMed), Embase, Web of Science, and the Cochrane Library for studies published in English from database inception to March 2022 that statistically compared the risk of psoriatic arthritis in patients aged > 16 years who were previously treated with biologic disease-modifying antirheumatic drugs or with other drugs for skin psoriasis. Four articles were eligible for analysis, all retrospective cohort studies. Three were conducted in preselected patients attending dermatology or dermatology-rheumatology collaboration centers and one was a large population-based study. In three studies, a primary two-step statistical analysis yielded a significantly lower risk of psoriatic arthritis in patients treated with biologic agents. These findings were not supported by the large retrospective electronic health record-based study. Biologic treatments may be effective in preventing the development of psoriatic arthritis in patients with psoriasis. More research is needed given the retrospective cohort design of all studies included in the review limiting the generalizability of the results, and the conflicting results from the registry study. At present, biologic agents should not be prescribed to unselected patients with psoriasis for the sole purpose of preventing psoriatic arthritis.


A nineteenth century view of the optic commissure.
Historically, ophthalmologists and anatomists have been intrigued by the optic nerve and its decussation. Isaac Newton was the first to describe and suggest the significance of the chiasmal crossing, while W.H. Wollaston defined the importance of hemiretinal differentiation in 1824. However, at that time, the vertical meridian separating nasal from temporal retina was believed to be located at the optic papillae. This theory was disproved in 1870 by two Philadelphia ophthalmologists, who demonstrated and documented the vertical meridian at the macula during their examination of an injured Civil War soldier.


Frontal brain injury chronically impairs timing behavior in rats.
Traumatic brain injury (TBI) affects over 2.8 million people annually, and has been shown to increase motor impulsivity in both humans and animals. However, the root cause of this behavioral disinhibition is not fully understood. The goal of the current study was to evaluate whether timing behavior is disrupted after TBI, which could potentially explain increases in impulsive responding. Twenty-one male three-month old Long-Evans rats were trained on a fixed interval-18 s schedule. Following training, rats were placed on the Peak Interval Procedure, with intermittent peak trials. On peak trials, no behaviors were reinforced and response rates were recorded to determine timing ability. After reaching a stable baseline, rats received bilateral frontal TBI (n = 12) using controlled cortical impact or sham procedures (n = 9). After one week recovery, rats were re-assessed on the Peak Procedure for six weeks. An amphetamine challenge was carried out after behavior reached stable post-injury performance. TBI caused a chronic decrease/acceleration in peak time, increase in response variability, and reduction in response rate. The shifted peak time suggests that altered perception of time may contribute to impairments in response inhibition after TBI. Amphetamine significantly increased response variability, with TBI animals demonstrating greater sensitivity, but did not affect peak time in either group. These data suggest that timing may not be the sole factor explaining impulsive action after TBI given that amphetamine reduced motor impulsivity in prior studies. Further investigations will be needed to dissociate the effects of amphetamine on TBI with regard to timing behavior.


Kinetic aspects of the interaction of blood clotting enzymes. VIII. The relation between clotting time and clotting velocity.
It is shown that in a system where thrombin acts on fibrinogen, the clotting time can be used to assess coagulation velocity because clotting time and clotting velocity are inversely proportional.


Small RNA-mediated repair of UV-induced DNA lesions by the DNA DAMAGE-BINDING PROTEIN 2 and ARGONAUTE 1.
As photosynthetic organisms, plants need to prevent irreversible UV-induced DNA lesions. Through an unbiased, genome-wide approach, we have uncovered a previously unrecognized interplay between Global Genome Repair and small interfering RNAs (siRNAs) in the recognition of DNA photoproducts, prevalently in intergenic regions. Genetic and biochemical approaches indicate that, upon UV irradiation, the DNA DAMAGE-BINDING PROTEIN 2 (DDB2) and ARGONAUTE 1 (AGO1) of Arabidopsis thaliana form a chromatin-bound complex together with 21-nt siRNAs, which likely facilitates recognition of DNA damages in an RNA/DNA complementary strand-specific manner. The biogenesis of photoproduct-associated siRNAs involves the noncanonical, concerted action of RNA POLYMERASE IV, RNA-DEPENDENT RNA POLYMERASE-2, and DICER-LIKE-4. Furthermore, the chromatin association/dissociation of the DDB2-AGO1 complex is under the control of siRNA abundance and DNA damage signaling. These findings reveal unexpected nuclear functions for DCL4 and AGO1, and shed light on the interplay between small RNAs and DNA repair recognition factors at damaged sites.


Metabolism of alkyldihydroxyacetone phosphate in rat brain.
Alkyldihydroxyacetone-P is the first detectable product in the biosynthetic pathway for ether-linked glycerolipids that eventually leads to the formation of ethanolamine plasmalogens, a major constituent of myelin. During early postnatal development, the specific activity of NADPH2:alkyldihydroxyacetone-P oxidoreductase in microsomes from rat brain is maximum at 4-5 days after birth, the time when the specific activity of the enzymes that synthesize alkyldihydroxyacetone-P also peaks. For the oxidoreductase assay, we developed a thin-layer chromatographic method that separates alkyldihydroxyacetone-P as the dinitrophenylhydrazine derivative from its reduced product (alkylglycerol-P), with excellent resolution. Phosphohydrolases associated with brain microsomes exhibit optimal pH maximums at 5.2-5.6 and 7.5-7.8 for all three substrates tested -- alkyldihydroxyacetone-P, alkylglycerol-P and alkylacylglycerol-P. Alkylglycerol-P was most readily dephosphorylated under all experimental conditions. The enzyme(s) that dephosphorylates alkyldihydroxyacetone-P and alkylglycerol-P have similar properties with respect to Mg-2+ or EDTA; with both substrates, Mg-2+ had no effect and EDTA was highly stimulatory. In contrast, EDTA strongly inhibited the dephosphorylation of alkylaclglycerol-P and although Mg-2+ (1 mM) appeared to be required for optimal activity, higher levels inhibited the reaction.


Development of a bifunctional crosslinking agent with potential for the preparation of immunotoxins.
A new protein crosslinking agent, 2,3-dibromopropionyl-N-hydroxysuccinimide ester, has been synthesized and characterized. The potential use of this compound as a temperature-controllable heterobifunctional crosslinking agent has been investigated using model systems and its reactivity compared with that of chlorambucil-N-hydroxysuccinimide ester. The coupling of 14C-labeled phenylethylamine to lysozyme has been used to illustrate the feasibility of the use of this crosslinking agent for the synthesis of immunotoxins.


Colon carcinogenesis models for chemoprevention studies.
Every animal model has its strengths and weaknesses; however, some of these models have proven useful in evaluating hereditary factors, whereas other models were found to be useful in understanding the relationship between nutritional factors and colon cancer. We believe that the results obtained by these models, both genetic based and chemically induced, will contribute to the understanding of genetic and nutritional factors as they relate to colon carcinogenesis.


Physical characterization of a stomatitis papulosa virus genome: a cleavage map for the restriction endonucleases HindIII and EcoRI.
The genome of stomatitis papulosa virus (a parapoxvirus) was cleaved with the restriction endonucleases HindIII and EcoRI, each giving rise to 6 fragments respectively. Double digestion with both enzymes resulted in 8 bands, two of which contained DNA fragments in double molar concentrations as revealed by reciprocal digests of isolated DNA fragments. The genome size, estimated by summation of the molecular weights of the fragments, is approximately 86 X 10(6) daltons, some 30 X 10(6) daltons smaller than vaccinia virus (an orthopoxvirus) DNA. The cleavage sites of HindIII and EcoRI endonucleases were mapped on the genome by analysis of reciprocal digests of isolated DNA fragments and by cross-hybridization experiments. This yielded two mapped segments which were then oriented relative to one another by cleavage of isolated partial digestion products. The terminal restriction fragments show rapid renaturation after alkali denaturation and subsequent neutralization, indicating that stomatitis papulosa virus DNA contains terminal cross-links analogous to those found in vaccinia virus DNA.


Long-term outcome of cervical and thoracolumbar dural arteriovenous fistulas with emphasis on sensory disturbance and neuropathic pain.
Clinical features and prognosis of sensory disturbance in spinal dural arteriovenous fistula (SDAVF) have not been well documented. Here we report long-term outcomes and detailed sensory evaluations of surgically treated SDAVF, including 14 patients with the craniocervical fistulas. Thirty-four consecutive patients with SDAVF treated at our institute during a period of 14 years were reviewed (mean age, 64.6 years; 67.6% men). Fistulas were located at the craniocervical junction in 14 patients (CC group) and in the thoracolumbar spine in 20 patients (TL group). In the CC group, six patients presented with subarachnoid hemorrhage. Fistulas were found incidentally in seven patients. One patient in the CC group and all patients in the TL group presented with progressive myelopathy. Most patients underwent microsurgery either alone (30 patients) or combined with embolization (3 patients). One patient was treated by embolization only. The follow-up ranged from 12 to 145 months (mean, 57 months). All but one patient in the CC group had excellent surgical outcome. Most patients in the TL group stabilized or improved neurologically. Shorter duration before treatment indicated better gait recovery. Important, a few patients in the TL group suffered worsening or development of new pain, as well as lesser degree of improvement in gait and micturition. Spinal cord atrophy was correlated with clinical deterioration. In craniocervical SDAVF, surgical treatment provides favorable long-term outcomes without risk of recurrence. In the thoracolumbar SDAVF, irreversible structural changes, such as spinal cord atrophy, may lead to poor recovery. Early diagnosis and treatment are thus warranted.


Unravelling colloid filter cake motions in membrane cleaning procedures.
The filtration performance of soft colloid suspensions suffers from the agglomeration of the colloids on the membrane surface as filter cakes. Backflushing of fluid through the membrane and cross-flow flushing across the membrane are widely used methods to temporally remove the filter cake and restore the flux through the membrane. However, the phenomena occurring during the recovery of the filtration performance are not yet fully described. In this study, we filtrate poly(N-isopropylacrylamide) microgels and analyze the filter cake in terms of its composition and its dynamic mobility during removal using on-line laser scanning confocal microscopy. First, we observe uniform cake build-up that displays highly ordered and amorphous regions in the cake layer. Second, backflushing removes the cake in coherent pieces and their sizes depend on the previous cake build-up. And third, cross-flow flushing along the cake induces a pattern of longitudinal ridges on the cake surface, which depends on the cross-flow velocity and accelerates cake removal. These observations give insight into soft colloid filter cake arrangement and reveal the cake's unique behaviour exposed to shear-stress.


HeatMapper: powerful combined visualization of gene expression profile correlations, genotypes, phenotypes and sample characteristics.
Accurate interpretation of data obtained by unsupervised analysis of large scale expression profiling studies is currently frequently performed by visually combining sample-gene heatmaps and sample characteristics. This method is not optimal for comparing individual samples or groups of samples. Here, we describe an approach to visually integrate the results of unsupervised and supervised cluster analysis using a correlation plot and additional sample metadata. We have developed a tool called the HeatMapper that provides such visualizations in a dynamic and flexible manner and is available from http://www.erasmusmc.nl/hematologie/heatmapper/. The HeatMapper allows an accessible and comprehensive visualization of the results of gene expression profiling and cluster analysis.


Treatment of sodium disorders after surgery of craniopharyngioma.
To study the treatment of the patients with blood sodium disorder after craniopharyngioma surgery. The blood sodium in 44 patients with craniopharyngiomas was daily examined from operative to post-operative days. Hypernatremia is defined as [Na] > 145 mmol/l, and hyponatremia as [Na] < 135 mml/L. Of the 44 patients, 36 developed sodium disorder. Among them, simple hyponatremia was 16 patients, simple hypernatremia was 9 patients, and alternative sodium disorder was 11 patients. The blood sodium disorder in patients with craniopharyngiomas after surgery appears to present complicated changes. Three types of blood sodium disorder could be determined as simple hyponatremia, simple hypernatremia, and alternative sodium disorder. Treatment of the sodium disorder in patients with after surgery could be carried in accordance with the above three types.


First-in-human autologous implantation of genetically modified adipocytes expressing LCAT for the treatment of familial LCAT deficiency.
Familial lecithin: cholesterol acyltransferase (LCAT) deficiency (FLD) is a severe inherited disease without effective treatment. Patients with FLD develop severe low HDL, corneal opacity, hemolytic anemia, and renal injury. We developed genetically modified adipocytes (GMAC) secreting LCAT (LCAT-GMAC) for ex vivo gene therapy. GMACs were prepared from the patient's adipocytes to express LCAT by retroviral gene transduction to secrete functional enzymes. This study aimed to evaluate the safety and efficacy of LCAT-GMAC implantation in an FLD patient. Proliferative preadipocytes were obtained from a patient using a ceiling culture and retrovirally transduced with LCAT. After obtaining enough cells by expansion culture of the transduced cells, the resulting LCAT-GMACs were implanted into a patient with FLD. To evaluate the safety and efficacy, we analyzed the outcome of the autologous implantation for 24 weeks of observation and subsequent 240 weeks of the follow-up periods. This first-in-human autologous implantation of LCAT-GMACs was shown to be safe by evaluating adverse events. The LCAT-GMAC implantation increased serum LCAT activity by approximately 50% of the baseline and sustained over three years. Consistent with increased LCAT activity, intermediate-density lipoprotein (IDL) and free cholesterol levels of the small and very small HDL fractions decreased. We found the hemoglobin/haptoglobin complex in the hemolyzed pre-implantation sera of the patient. After one week of the implantation, the hemoglobin/haptoglobin complex almost disappeared. Immediately after the implantation, the patient's proteinuria decreased temporarily to mild levels and gradually increased to the baseline. At 48 weeks after implantation, the patient's proteinuria deteriorated with the development of mild hypertension. By the treatment with antihypertensives, the patient's blood pressure normalized. With the normalization of blood pressure, the proteinuria rapidly decreased to mild proteinuria levels. LCAT-GMAC implantation in a patient with FLD is shown to be safe and appears to be effective, in part, for treating anemia and proteinuria in FLD.


Studies on the mechanism of ethanol-induced gastric damage in rats.
Concentrated ethanol causes gastric lesions by a mechanism that is poorly understood. We have investigated this mechanism in the rat stomach via gross morphologic, videomicroscopic, histochemical, and pharmacologic approaches. Within 1 min of contact, ethanol caused diffuse mucosal hyperemia. By 5 min, hyperemia greatly intensified at some mucosal sites. Beneath sites where mucosal hyperemia developed, intramural venules strongly constricted at 3-13 s postethanol, whereas submucosal arterioles dilated more than two times in diameter by 25 s. Submucosal venular constriction began sooner than arteriolar dilation (9 vs. 16 s, p less than 0.05). One-third of the gastric mucosal mast cells degranulated by 15 s postethanol; 50% discharged by 30 s. Ethanol-induced hyperemia was markedly reduced by lipoxygenase-selective inhibitors BW755C or nordihydroguaiaretic acid, or by the H1-antihistamine pyrilamine, but not by indomethacin, cimetidine, phentolamine, or methysergide. Based on these results, a model for the pathogenesis of ethanol-induced gastric lesions is proposed.


Different means, same end-heterochromatin formation by RNAi and RNAi-independent RNA processing factors in fission yeast.
The assembly of heterochromatin in eukaryotic genomes is critical for diverse chromosomal events including regulation of gene expression, silencing of repetitive DNA elements, proper segregation of chromosomes and maintenance of genomic integrity. Previous studies have shown that noncoding RNAs and the RNA interference (RNAi) machinery promote the assembly of heterochromatin that serves as a multipurpose platform for targeting effectors involved in various chromosomal processes. Recent work has revealed that RNAi-independent mechanisms, involving RNA processing activities that utilize both noncoding and coding RNAs, operate in the assembly of heterochromatin. These findings have established that, in addition to coding for proteins, mRNAs also function as signaling molecules that modify chromatin structure by targeting heterochromatin assembly factors.


Atopic Dermatitis: A Qualitative Study on the Burdens of Living with Itchiness.
We sought to understand the consequences itchiness has on daily life that may not be immediately obvious in clinical assessments for patients with atopic dermatitis (AD). Focus groups and interviews involving 21 patients with AD and 12 family members examined aspects of the effects of itchiness on health-related quality of life (HRQL). Investigators conducted a thematic analysis where two researchers independently coded the narratives and arrived at a consensus on major themes. Five themes emerged from our discussions. 1) Miserable experience: Itchiness was difficult to control and cease. 2) Physical damage: Damage to skin and hair occurred from scratching to alleviate the itchiness. 3) Effects on daily activities: Itchiness could affect everything participants did, including how they dressed, used make-up, and slept. 4) Effects on social activities and relationships: The discomfort and embarrassment from scratching in public and others' reactions hindered participants' social lives. 5) Emotional consequences: Various emotional responses to itchiness were reported, including embarrassment, depression, and irritation. Though qualitative research provides a level of detail not often found in quantitative analyses, this study design is limited by small sample size and generalizability. Understanding these challenges can help clinicians open deeper conversations with their patients to learn more about what patients need from their dermatologic care. While itchiness from AD is well-known, this study shows that its effects on HRQL are not minimal and that patients may need further care for the consequences of this symptom.


[Rhinitis].
Acute rhinitis is an acute inflammatory disease affecting nasal mucous membranes, most frequently caused by viral infections. Complications include secondary bacterial invasion. Treatment is mainly symptomatic. Antibiotics should be prescribed only when significant bacterial infection is demonstrated. Local vasoactive drugs should not be used but occasionally their oral administration can be effective. Hypertrophic and atrophic forms may be differentiated among chronic rhinitis. Hypertrophic forms include congestive, hiperplastic and allergic lesions of nasal and sinus mucous membranes. Allergic rhinitis should be treated with antihistamines, descongestants and avoidance of allergen. The atrophic forms with crust formation and foul breath are usually managed with local flushings of 9% sodium chloride in water followed by instillation of fluid vaseline, but in some resistant cases reduction of intranasal space by surgical insertion of acrylic pieces is recommended. Nasal obstruction, foul odor, dryness sensation and headache are usually controlled in 90% of these surgically treated patients.


Synthesis and biological evaluation of indolyl glyoxylamides as a new class of antileishmanial agents.
A series of indolylglyoxylamide derivatives have been synthesized and evaluated in vitro against amastigote form of Leishmania donovani. Compound 8c has been identified as the most active analog of the series with IC(50) value of 5.17μM and SI value of 31.48, and is several folds more potent than the standard drugs sodium stilbogluconate and pentamidine.


A systematic review and meta-analysis of randomised trials of neo-adjuvant hormone therapy for localised and locally advanced prostate carcinoma.
We performed a systematic review and meta-analysis of randomised trials of neo-adjuvant hormone therapy (NHT) in localised and locally advanced prostate cancer to assess the effectiveness of this therapy. We searched MEDLINE, The Cochrane Library, Science Citation Index, LILACS and SIGLE for randomised trials comparing NHT plus primary therapy (radiotherapy or prostatectomy) with primary therapy alone. Data included information on study design, participants, interventions, and outcomes. Comparable data were extracted from eligible studies and pooled for meta-analysis with intention to treat principle. NHT prior to prostatectomy did not improve overall or disease-free survival, but did significantly reduce positive margin rates (RR 0.49, 95% CI 0.42-0.56, p<0.00001), organ confinement (RR 1.63, 95% CI 1.37-1.95, p<0.0001) and lymph node invasion (RR 0.49, 95% CI 0.42-0.56, p<0.02). In one study NHT before radiotherapy significantly improved overall survival for men with Gleason 2-6 (p=0.015). In addition, there was a significant improvement in both clinical disease-free survival (RR 1.46, 95% CI 1.24-1.71, p<0.00001) and biochemical disease-free survival (RR 1.59, 95% CI 1.00-2.55, p=0.05). Toxicities included hot flushes, gastrointestinal, hepatic and miscellaneous adverse events. NHT is associated with significant clinical benefit when given with radiotherapy and improves pathological outcome prior to prostatectomy but is of minimal value prior to radical prostatectomy. The decision to use hormone therapy should be discussed between the patient, the clinician and policy maker based on the benefits, toxicity and cost.


Familial hereditary progressive sensorineural hearing loss among Saudi population.
Clinical and audiological studies of 234 Saudi patients from the central area of Saudi Arabia with progressive sensorineural hearing loss were carried out in Riyadh. One hundred and sixty-four came from 38 families, i.e. 70% of the total and 30% were sporadic cases. Consanguinity was found in 80.8%. Their hearing loss was characterized as being bilateral sensorineural, starting at 1 kHz frequency and gradually sloping the pure tone audiogram to severe and profound hearing loss at 8 kHz. Special tests for recruitment indicate hair cell lesions. Caloric tests were normal in 56 patients tested. No abnormal internal ears were found in a C.T. scan done for 84 cases. Serology for TORCH and non-TORCH agents were done for (124) selected cases. Syphilis was negative, however, positive immunoglobulins for cytomegalovirus, herpes simplex type 1 and 2, and toxoplasmosis were seen in 52 cases. All patients came from one area called Qassim where old customs and the tradition of consanguineous marriage were still practiced. Some of these families are now under molecular and cytogenetic study.


[Not Available].
An analytical method for the detection of fusaric acid in wheat and wheat products has been developed as well as employed for the survey of randomly selected samples (n=33). While the application of GC-FID proved to be not sensitive enough for this purpose, the sensitivity and selectivity of GC-MS permitted the quantification of the mycotoxin in 78% of the wheat samples.The measured concentrations ranged from 7 to 570 μg/kg (average value 134 μg/kg). However, clear differences were recognizable regarding the type of sample: While field samples showed 332 μg/kg on an average, cleaned grains designated for human consumption contained averaged 138 μg/kg, and wholemeal products 68 μg/kg.Although the limited number of samples does not permit final conclusions, the toxicological relevance of fusaric acid alone may probably be classified as small. To what extent the well known cumulative effects with deoxynivalenol and fumonisin B1 impair this evaluation has to remain elusive.


[Fatal decompression sickness following therapeutic hyperbaric environment (author's transl)].
In Hannover/Germany in 1976 a so called "Society for Regenerative Hyperbaric Therapy" (Gesellschaft für Regenerative Uberdruck-Therapie) subjected 20 patients within two coupled multi-place chambers to a simulated hyperbaric environment, equivalent to a maximum of 4 ata, followed usually by gradual reductions of the pressure. The patients were of an average age of 67.2 years and were afflicted by various disorders. During one of the "dives" a patient developed air embolism of the central nervous system. During ensuing confusion of the chamber was opened suddenly, with resultant explosive reduction of the high pressure. This resulted in five letal accidents of decompression sickness. Post mortem examination revealed diffuse distribution of gas bubbles throughout the entire body. By elecronmicroscopy each bubble was covered by an osmiophilic coat. Post mortem findings are discussed and correlated with well-known and new clinical symptomes. These observations present for the first time fatal accidents of decompression sickness in humans, associated with hyperbaric air-therapy.


Molecular weight determination of lignosulfonates by size-exclusion chromatography and multi-angle laser light scattering.
A lignosulfonate sample was fractionated according to the solubility in ethanol-water. The fractions were analysed by aqueous size exclusion chromatography (SEC) combined with in-line multi-angle laser light scattering (MALLS), and by static MALLS. Satisfactory SEC results were obtained with aqueous phosphate buffer containing DMSO and SDS. The refractive index increment (dn/dc) varied from 0.186 to 0.205 ml/g, depending on Mw and the degree of sulfonation. The second viral coefficient (A2) was 7 x 10(-3) ml mol/g2. The weight-average molecular weight (Mw) of the fractions varied from 4600 up to 398 000 g/mol. and the polydispersity (Mw/Mn) varied between 1.3 and 3.5.


Depicting the interplay between organisational tiers in the use of a national quality registry to develop quality of care in Sweden.
With a pending need to identify potential means to improved quality of care, national quality registries (NQRs) are identified as a promising route. Yet, there is limited evidence with regards to what hinders and facilitates the NQR innovation, what signifies the contexts in which NQRs are applied and drive quality improvement. Supposedly, barriers and facilitators to NQR-driven quality improvement may be found in the healthcare context, in the politico-administrative context, as well as with an NQR itself. In this study, we investigated the potential variation with regards to if and how an NQR was applied by decision-makers and users in regions and clinical settings. The aim was to depict the interplay between the clinical and the politico-administrative tiers in the use of NQRs to develop quality of care, examining an established registry on stroke care as a case study. We interviewed 44 individuals representing the clinical and the politico-administrative settings of 4 out of 21 regions strategically chosen for including stroke units representing a variety of outcomes in the NQR on stroke (Riksstroke) and a variety of settings. The transcribed interviews were analysed by applying The Consolidated Framework for Implementation Research (CFIR). In two regions, decision-makers and/or administrators had initiated healthcare process projects for stroke, engaging the health professionals in the local stroke units who contributed with, for example, local data from Riksstroke. The Riksstroke data was used for identifying improvement issues, for setting goals, and asserting that the stroke units achieved an equivalent standard of care and a certain level of quality of stroke care. Meanwhile, one region had more recently initiated such a project and the fourth region had no similar collaboration across tiers. Apart from these projects, there was limited joint communication across tiers and none that included all individuals and functions engaged in quality improvement with regards to stroke care. If NQRs are to provide for quality improvement and learning opportunities, advances must be made in the links between the structures and processes across all organisational tiers, including decision-makers, administrators and health professionals engaged in a particular healthcare process.


Ultrafast cold-brewing of coffee by picosecond-pulsed laser extraction.
Coffee is typically brewed by extracting roasted and milled beans with hot water, but alternative methods such as cold brewing became increasingly popular over the past years. Cold-brewed coffee is attributed to health benefits, fewer acids, and bitter substances. But the preparation of cold brew typically needs several hours or even days. To create a cold-brew coffee within a few minutes, we present an approach in which an ultrashort-pulsed laser system is applied at the brewing entity without heating the powder suspension in water, efficiently extracting caffeine and aromatic substances from the powder. Already 3 min irradiation at room temperature leads to a caffeine concentration of 25 mg caffeine per 100 ml, comparable to the concentrations achieved by traditional hot brewing methods but comes without heating the suspension. Furthermore, the liquid phase's alkaloid content, analyzed by reversed-phase liquid chromatography coupled to high-resolution mass spectrometry, is dominated by caffeine and trigonelline and is comparable to traditional cold-brewed coffee rather than hot-brewed coffee. Furthermore, analyzing the head-space of the prepared coffee variants, using in-tube extraction dynamic head-space followed by gas chromatography coupled to mass spectrometry, gives evidence that the lack of heating leads to the preservation of more (semi-)volatile substances like pyridine, which provide cold-brew coffee its unique taste. This pioneering study may give the impetus to investigate further the possibility of cold-brewing coffee, accelerated by more than one order of magnitude, using ultrafast laser systems.


Nonoperative management of biliary ductal stenosis. Report of a most unusual case.
A patient with documented iatrogenic stricture of the common duct is reported. Certain very unusual characteristics of this lesion suggested that it was not precisely similar to ordinary biliary strictures and that nonoperative care was worthy of trial. No reconstructive operation has been done. The strictured segment has resolved so that the patient is completely asymptomatic and has no direct or indirect laboratory or radiographic evidence of biliary obstruction.


Vitamin D Deficiency Accelerates Coronary Artery Disease Progression in Swine.
The role of vitamin D deficiency in coronary artery disease (CAD) progression is uncertain. Chronic inflammation in epicardial adipose tissue (EAT) has been implicated in the pathogenesis of CAD. However, the molecular mechanism underlying vitamin D deficiency-enhanced inflammation in the EAT of diseased coronary arteries remains unknown. We examined a mechanistic link between 1,25-dihydroxyvitamin D-mediated suppression of nuclear factor-κB (NF-κB) transporter, karyopherin α4 (KPNA4) expression and NF-κB activation in preadipocytes. Furthermore, we determined whether vitamin D deficiency accelerates CAD progression by increasing KPNA4 and nuclear NF-κB levels in EAT. Nuclear protein levels were detected by immunofluorescence and Western blot. Exogenous KPNA4 was transported into cells by a transfection approach and constituted lentiviral vector. Swine were administered vitamin D-deficient or vitamin D-sufficient hypercholesterolemic diet. After 1 year, the histopathology of coronary arteries and nuclear protein expression of EAT were assessed. 1,25-dihydroxyvitamin D inhibited NF-κB activation and reduced KPNA4 levels through increased vitamin D receptor expression. Exogenous KPNA4 rescued 1,25-dihydroxyvitamin D-dependent suppression of NF-κB nuclear translocation and activation. Vitamin D deficiency caused extensive CAD progression and advanced atherosclerotic plaques, which are linked to increased KPNA4 and nuclear NF-κB levels in the EAT. 1,25-dihydroxyvitamin D attenuates NF-κB activation by targeting KPNA4. Vitamin D deficiency accelerates CAD progression at least, in part, through enhanced chronic inflammation of EAT by upregulation of KPNA4, which enhances NF-κB activation. These novel findings provide mechanistic evidence that vitamin D supplementation could be beneficial for the prevention and treatment of CAD.


Effect of fibrillation on acetylcholinesterase mRNA in cultured embryonic rat myotubes.
Acetylcholinesterase (AChE) and AChE mRNA were evaluated in spontaneously fibrillating myotubes derived from 20-day-old rat fetuses and in matched cultures in which fibrillation was prevented by adding tetrodotoxin on the fourth day of culture. On the eighth day of culture, the AChE activity of fibrillating and nonfibrillating cultures was 5332 and 1861 pmol ACh hydrolyzed min-1 dish-1, respectively (P less than 0.005). Total mRNA was essentially the same in fibrillating and nonfibrillating cultures (27.4 and 25.4 micrograms/dish, respectively). AChE mRNA was assessed by assaying the AChE produced by Xenopus oocytes microinjected with purified mRNA. The AChE produced by mRNA from fibrillating and nonfibrillating cultures was 0.46 and 0.10 pmol ACh hydrolyzed min-1 oocyte-1, respectively (P less than 0.005).


Use of ACTH and prednisolone in infantile spasms: experience from a developing country.
Adrenocorticotrophic hormone (ACTH) and prednisone are both used to treat infantile spasms (IS) in West syndrome. In many countries, ACTH is expensive and difficult to obtain whereas, prednisone or prednisolone are cheap, given orally and easily available. The purpose of this retrospective data analysis was to compare the efficacy and cost of ACTH and prednisolone in the treatment of IS from the perspective of a developing country. Patients admitted with West syndrome in Children's Hospital, Islamabad, between January 1995 and December 2001 were included in the analysis. The diagnosis was made after eliciting a history of characteristic seizures and detecting hypsarrhythmia on the EEG. Parents were offered the use of either ACTH administered by intramuscular injection or prednisolone given orally. ACTH was expensive and difficult to obtain whereas prednisolone was cheap and easily available. One hundred and five children were included in the study. Sixty-three were boys and their age ranged from 2 months to 3 years with a mean of 11 months. Thirty-three children received ACTH injections; 27 showed improvement and 11 remained spasms free after discontinuation of injections. Seventy-two patients were given oral prednisolone, 51 responded and 17 remained spasms free after oral steroids were stopped. Overall outcome was similar in both groups. The cost of ACTH injection was more than 100 times the cost of oral prednisolone. No significant difference was seen in the final outcome in both treatment groups. Since prednisolone is inexpensive, easily available and given orally, it is the preferred mode of therapy.


Improved antioxidant effect of idebenone-loaded polyethyl-2-cyanoacrylate nanocapsules tested on human fibroblasts.
The protective antioxidant role of idebenone both as free drug and drug-loaded Tween 80-coated polyethyl-2-cyanoacrylate (PECA) nanocapsules is reported. The relationship between oxidative damage and apoptotic or nonapoptotic cell death is evaluated in vitro. Idebenone-loaded nanocapsules were prepared with the interfacial polymerization method in the presence of Tween 80. Human nonimmortalized fibroblasts. under different stress conditions, either 0.5 mM diethylmaleate (DEM) for 60 min or 0.1 mM H2O2 for 30 min, were used as the experimental in vitro model. The production of reactive oxygen species, the cell viability, and the nuclear DNA damage were evaluated. The presence of apoptotic damage was evaluated both by the determination of caspase-3-like protein activity and by Promega's fluorescent apoptotic detection system. DEM and H2O2 affected the cultured cells in different ways. DEM induced a moderate cellular insult, which was efficaciously antagonized by idebenone-loaded PECA nanocapsules. H2O2 elicited severe damage to nuclear DNA, which was reduced by idebenoneloaded PECA nanocapsules. The free drug was less effective than idebenone-loaded nanocapsules. The findings reported here demonstrate that an improved antioxidant effect was obtained with a low idebenone concentration (0.5 microM) when the drug was entrapped within Tween 80-coated PECA nanocapsules.


Level of exclusive breastfeeding and its enabling factors among lactating women who delivered in health facilities of Asosa town, Ethiopia: a cross sectional study.
Despite the enormous benefit of exclusive breastfeeding (EBF) to mothers and infants, the practice of exclusive breastfeeding is globally low. In sub-Saharan Africa and Ethiopia, the prevalence of EBF stands at 35% and 59%, respectively. The low EBF practice in Ethiopia as well as in the studied region calls for further study and thus we studied the EBF practice in the study area since little is known about its current magnitude and factors influencing its practice for some programmatic improvements. A facility based cross-sectional study linked to a community was conducted from June-July 2019 among 412 mothers who had antenatal follow-up and delivered in health facilities of Asosa town over a period of one year prior to the study. Data on socio demographic characteristics and other important variables were collected through face to face interview while gestational age in weeks was recorded from their medical chart by trained health extension workers in accordance with relevant ethical guidelines and regulations. The collected data were then cleaned and entered into Epi-data software version 3.02. Analysis was done by SPSS version 20. Binary and multivariate logistic regression were performed to identify the contributing factors. P-value of less than 0.05 and 95% confidence interval was considered to determine statistical significance. Of the 412 respondents, the majority (88.1%) were multi-gravida and above. Slightly higher than a quarter (26.0%) and over half (55.4%) had neither received antenatal nor postnatal care. The proportion of mothers who exclusively breastfed their children was 76.0% and the overall aggregated good practice of EBF score was 64.1%. Mothers who completed primary school [AOR = 4.5; 95% CI = 1.1,18.2], had four or more ANC [AOR = 1.8;95CI = 0.79-0.98], and postnatal follow-up [AOR = 0.21;95% CI = 0.07-0.67], and had male infants [AOR = 2.3; 95% CI = 1.0-4.95] were among the factors influencing the exclusive breastfeeding practice score. While three in four mothers exclusively breastfed their newborns and about two-thirds had good EBF score, the observed women's retention on the continuum of the maternal care pathway is low with one in four had no antenatal and over half had no postnatal care which are important derivers for EBF practice. To improve the EBF score and narrow the observed maternal and child health disparities, it is essential to promote maternal education and increase the recomended coverage of antenatal and postnatal care for mothers.


Erratum: In vitro assessment of pesticides capacity to act as agonists/antagonists of the thyroid hormone nuclear receptors.
[This corrects the article DOI: 10.1016/j.isci.2021.102957.].


Non-specific cross-reacting antigen (NCA) in individual maturation stages of myelocytic cell series.
The distribution and localization of NCA and carcinoembryonic antigen CEA in cells of different types of myelogenous leukaemias (acute myelogenous leukaemia - AML; chronic granulocytic leukaemia - CGL; CGL in myeloblastic crisis - CGL-BC) was studied using the immunofluorescence test. Discontinuous density-gradient centrifugation was used to separate myeloid cells into fractions containing granulocytes in individual stages of maturation. Serum NCA and CEA levels were estimated in parallel. It was established that: (a) AML blasts without maturation (MO type) and monoblasts did not synthesize NCA; (b) individual blasts of AML with features of maturation (M1, M2 types) and some myeloblasts of CGL-BC exhibited a limited ability to express cytoplasmic NCA; (c) the number of NCA-containing cells increased in the more mature granulocyte fractions isolated on Ficoll-Hypaque density-gradients; (d) myelocytic NCA is immunologically related to NCA isolated from lung tissue and (e) CEA is undetectable in the myelocytic cell series.


[A further study on potential of microsatellite analysis of urine in diagnosis of bladder cancer].
To investigate the application value of microsatellite analysis of voided urine in diagnosis of bladder cancer and compare the advantages and disadvantages of this method and urine cytology. Thirteen microsatellite markers, including the nine markers used in the authors' previous study and another four new markers, were tested on urine DNA from 143 individuals with or without bladder lesions in a blinded fashion. Urine cytology was applied at the same time. The urine DNA from 89% (97/109) of the patients with bladder cancer, 86% (6/7) of the patients with inverted papilloma of the bladder, 3 out of 7 patients with cystitis and one patient with hematuria of unknown cause showed alterations, at least, in one of the 13 microsatellite markers, while the 19 individuals without bladder lesion had no change at any of these loci. Nucleic atypia or suspicious cancer cells were detected by routine cytology in the urine of the 3 patients with cystitis and one patient with hematuria of unknown cause who showed alterations of microsatellite in their urine sediments. Among the 81 cases of bladder cancer whose urine was examined by both microsatellite analysis and routine cytology, 73 cases (90%), of various stage or grade, were identified by microsatellite analysis, while only 41 cases (51%), mainly at advanced stage, were detected by cytology. Cancer cells were detected in the urine sediments of 4 of the 12 cases that were not identified by microsatellite analysis. Microsatellite analysis of urine is useful for detection of bladder cancer. Microsatellite analysis and urine cytology could be complementary to each other.