Platelet aggregation in acute coronary syndromes: use of a new aggregometer with laser light scattering to assess platelet aggregability.
Platelet aggregation has been implicated in the pathogenesis of acute coronary syndromes. Small aggregates consisting of < or = 100 platelets cannot be quantified with a conventional aggregometer employing optical density. Using a recently developed aggregometer based on laser light scattering, we studied platelet aggregability in patients with acute coronary syndromes. Peripheral blood samples were obtained from 39 patients with acute myocardial infarction or unstable angina who had received no prior antiplatelet or anticoagulant therapy, to be assayed immediately using a PA-100 platelet aggregometer. Blood samples from 14 healthy volunteers were used as controls. Spontaneous formation of platelet aggregates was observed only in patients with acute coronary syndromes. The size of these aggregates was small, consisting of < or = 100 platelets (primary aggregation). Agonist-induced aggregation consisted of two phases. In the first few minutes, the number of small aggregates increased markedly (primary aggregation), followed by an increase in larger aggregates (secondary aggregation). The EC50 of epinephrine for primary aggregation was nearly 50 times lower in acute coronary patients than in controls (P < 0.001), while the EC50 for secondary aggregation was only 2 times lower (P < 0.001). Aggregometry using light scattering suggests that platelet hyperaggregability and hypersensitivity in acute coronary syndromes may occur in primary but not secondary aggregation.


Machine learning methods for functional recovery prediction and prognosis in post-stroke rehabilitation: a systematic review.
Rehabilitation medicine is facing a new development phase thanks to a recent wave of rigorous clinical trials aimed at improving the scientific evidence of protocols. This phenomenon, combined with new trends in personalised medical therapies, is expected to change clinical practice dramatically. The emerging field of Rehabilomics is only possible if methodologies are based on biomedical data collection and analysis. In this framework, the objective of this work is to develop a systematic review of machine learning algorithms as solutions to predict motor functional recovery of post-stroke patients after treatment. We conducted a comprehensive search of five electronic databases using the Patient, Intervention, Comparison and Outcome (PICO) format. We extracted health conditions, population characteristics, outcome assessed, the method for feature extraction and selection, the algorithm used, and the validation approach. The methodological quality of included studies was assessed using the prediction model risk of bias assessment tool (PROBAST). A qualitative description of the characteristics of the included studies as well as a narrative data synthesis was performed. A total of 19 primary studies were included. The predictors most frequently used belonged to the areas of demographic characteristics and stroke assessment through clinical examination. Regarding the methods, linear and logistic regressions were the most frequently used and cross-validation was the preferred validation approach. We identified several methodological limitations: small sample sizes, a limited number of external validation approaches, and high heterogeneity among input and output variables. Although these elements prevented a quantitative comparison across models, we defined the most frequently used models given a specific outcome, providing useful indications for the application of more complex machine learning algorithms in rehabilitation medicine.


Development of a promoter shutoff system in Aspergillus oryzae using a sorbitol-sensitive promoter.
Promoter shutoff is a general method for analyzing essential genes, but in the fungus Aspergillus oryzae, no tightly repressed promoters have been reported. To overcome the current limitations of conditional promoters, we examined sorbitol- and galactose-responsive genes using microarrays to identify regulatable genes with only minor physiological and genetic effects. We identified two sorbitol-induced genes (designated as sorA and sorB), cloned their promoters, and built a regulated egfp and brlA expression system. Growth medium-dependent enhanced green fluorescence protein (EGFP) fluorescence and conidiation were confirmed for egfp and brlA under the control of their respective promoters. We also used this shutoff system to regulate the essential rhoA, which demonstrated the expected growth inhibition under repressed growth conditions. Our new sorbitol promoter shutoff system developed can serve as a valuable new tool for essential gene analyses of filamentous fungi.


Accuracy of intraoperative frozen-section analysis of breast cancer lumpectomy-bed margins.
My colleagues and I have been using intraoperative frozen-section analysis (FSA) to evaluate lumpectomy margins in an attempt to reduce the number of additional operations that patients with ductal carcinoma in situ or stage I and II breast cancer would have to endure. We review our experience in breast-conservation therapy (BCT) at the University of Florida (Gainesville) to determine the effectiveness of this approach. Operative reports, operative logs, and pathology reports were retrospectively reviewed for patients who had BCT from January 2001 to January 2004. Ninety-seven patients (116 operations) were reviewed. Nineteen patients required an additional operation (19.6%). Forty-three patients had positive margins on paraffin-embedded histologic analysis (44.3%). Accuracy of FSA was 84% when evaluated on a per-case basis, and 96% on a per-slide basis. False negatives were identified in 22 patients, affecting the operative pathway of 19 patients (19.6%) and were identified more frequently in cases of ductal carcinoma in situ (p < 0.001). There were no false positives. Additional operative time required for FSA was approximately 13 minutes per case. Eighty-four (86.6%) patients had successful BCT and 13 patients (13.4%) required mastectomy. Intraoperative analysis of margins using FSA is effective at minimizing the number of additional operations, with 19 patients benefiting from immediate intervention in this study. The authors believe that the number of second operations prevented and the high BCT rates justify performing FSA. Ductal carcinoma in situ is more difficult to identify in FSA. Preoperative discussions with the patient should reflect these findings.


Impact of Catheter Ablation on Sleep Quality and Relationship Between Sleep Stability and Recurrence of Paroxysmal Atrial Fibrillation After Successful Ablation: 24-Hour Holter-Based Cardiopulmonary Coupling Analysis.
Background Sleep fragmentation and sleep apnea are common in patients with atrial fibrillation (AF). We investigated the impact of radio-frequency catheter ablation (RFCA) on sleep quality in patients with paroxysmal AF and the effect of a change in sleep quality on recurrence of AF. Methods and Results Of 445 patients who underwent RFCA for paroxysmal AF between October 2007 and January 2017, we analyzed 225 patients who had a 24-hour Holter test within 6 months before RFCA. Sleep quality was assessed by cardiopulmonary coupling analysis using 24-hour Holter data. We compared cardiopulmonary coupling parameters (high-frequency coupling, low-frequency coupling, very-low-frequency coupling) before and after RFCA. Six months after RFCA, the high-frequency coupling (marker of stable sleep) and very-low-frequency coupling (rapid eye movement/wake marker) was significantly increased (29.84%-36.15%; P<0.001; and 26.20%-28.76%; P=0.002, respectively) while low-frequency coupling (unstable sleep marker) was decreased (41.25%-32.13%; P<0.001). We divided patients into 3 tertiles according to sleep quality before RFCA, and the risk of AF recurrence in each group was compared. The second tertile was used as a reference; patients with unstable sleep (Tertile 3) had a significantly lower risk of AF recurrence (hazard ratio [HR], 0.32; 95% CI, 0.12-0.83 for high-frequency coupling; and HR, 0.22; 95% CI, 0.09-0.58 for low-frequency coupling). Conclusions Sleep quality improved after RFCA in patients with paroxysmal AF. The recurrence rate was significantly lower in patients who had unstable sleep before RFCA. These results suggest that RFCA can influence sleep quality, and sleep quality assessment before RFCA may provide a risk marker for recurrence after RFCA in patients with paroxysmal AF.


MTP18 inhibition triggers mitochondrial hyperfusion to induce apoptosis through ROS-mediated lysosomal membrane permeabilization-dependent pathway in oral cancer.
Although stress-induced mitochondrial hyperfusion (SIMH) exerts a protective role in aiding cell survival, in the absence of mitochondrial fission, SIMH drives oxidative stress-related induction of apoptosis. In this study, our data showed that MTP18, a mitochondrial fission-promoting protein expression, was increased in oral cancer. We have screened and identified S28, a novel inhibitor of MTP18, which was found to induce SIMH and subsequently trigger apoptosis. Interestingly, it inhibited MTP18-mediated mitochondrial fission, as shown by a decrease in p-Drp1 along with increased Mfn1 expression in oral cancer cells. Moreover, S28 induced autophagy but not mitophagy due to the trouble in engulfment of hypoperfused mitochondria. Interestingly, S28-mediated SIMH resulted in the loss of mitochondrial membrane potential, leading to the consequent generation of mitochondrial superoxide to induce intrinsic apoptosis. Mechanistically, S28-induced mitochondrial superoxide caused lysosomal membrane permeabilization (LMP), resulting in decreased lysosomal pH, which impaired autophagosome-lysosome fusion. In this setting, it showed that overexpression of MTP18 resulted in mitochondrial fission leading to mitophagy and inhibition of superoxide-mediated LMP and apoptosis. Further, S28, in combination with FDA-approved anticancer drugs, exhibited higher apoptotic activity and decreased cell viability, suggesting the MTP18 inhibition combined with the anticancer drug could have greater efficacy against cancer.


Differentiation of primary embryonic neuroblasts in purified neural cell cultures from Drosophila.
Embryonic neuroblasts of Drosophila are undifferentiated precursor cells that give rise to the central nervous system. Centrifugal elutriation has been employed to fractionate embryonic cells on the basis of size. A fraction of large cells was found to be greatly enriched for neuroblasts, whereas mesodermal precursor cells were completely excluded. This allowed a second step of purification, based upon adhesion to glass, to provide virtually pure cultures of neural cells. The cells in these cultures had the properties of neurons of the Drosophila CNS: They gave rise to ganglion-like clusters from which neurites extended on the culture substrate, and they expressed the enzyme, acetylcholinesterase, and the cell surface antigens recognized by antisera raised against horseradish peroxidase. The production of large-scale neuronal cell cultures will be useful for immunological and molecular studies of neural cell differentiation.


Promoting sustainable responses to the US opioid epidemic with community-academic partnerships: qualitative outcomes from a statewide program.
Drug overdose deaths in the United States have continued to increase at an alarming rate. The Substance Abuse and Mental Health Services Administration (SAMHSA) distributed more than $7 billion between January 2016 and June 2020 to address the drug overdose crisis. The funds support evidence-based responses, including medications for opioid use disorder, and other prevention, treatment and recovery activities. Although the State Opioid Response (SOR) grants finance much-needed community level interventions, many of the services they support may not be sustainable, without ongoing assessment, evaluation and planning for continuation. This paper describes a statewide effort to support local entities through SAMHSA's SOR grants in Virginia. Community agencies across the state participated in detailed needs assessment exercises with VHEOC investigators, and developed requests for proposals (RFPs) to sustain their SOR programs. The RFPs were then distributed to prospective academic partners at the five VHEOC universities, based on the required subject matter expertise identified in the RFP. All responsive proposals were then provided to the local agencies who selected the proposal most likely to meet their needs. VHEOC investigators also conducted an inductive, three-phase content analysis approach to examine the RFPs submitted to the VHEOC to identify nominal categories of support requested of the VHEOC investigators. VHEOC Investigators received and coded 27 RFPs from ten community agencies representing four of five regions of the state. We identified six nominal categories of academic assistance with high inter-coder agreement. The six categories of support requested of the academic partners were program development and support, literature review and best practices, outreach and education, data analysis and interpretation, program evaluation, and grant writing assistance. Several RFPs requested up to three categories of support in a single project. Our analysis of the requests received by the consortium identified several categories of academic support for SOR-grantees addressing the drug overdose crisis. The most common requests related to development and maintenance of supportive collaborations, which existing research has demonstrated is necessary for the long-term sustainability of SOR-funded services. In this way, the academic partners reinforced sustainable SOR-funded programs. As the state opioid response program is implemented nationally, we hope that other states will consider similar models in response to the opioid crisis.


Long Duration of Ground Motion in the Paradigmatic Valley of Mexico.
Built-up on top of ancient lake deposits, Mexico City experiences some of the largest seismic site effects worldwide. Besides the extreme amplification of seismic waves, duration of intense ground motion from large subduction earthquakes exceeds three minutes in the lake-bed zone of the basin, where hundreds of buildings collapsed or were seriously damaged during the magnitude 8.0 Michoacán earthquake in 1985. Different mechanisms contribute to the long lasting motions, such as the regional dispersion and multiple-scattering of the incoming wavefield from the coast, more than 300 km away the city. By means of high performance computational modeling we show that, despite the highly dissipative basin deposits, seismic energy can propagate long distances in the deep structure of the valley, promoting also a large elongation of motion. Our simulations reveal that the seismic response of the basin is dominated by surface-waves overtones, and that this mechanism increases the duration of ground motion by more than 170% and 290% of the incoming wavefield duration at 0.5 and 0.3 Hz, respectively, which are two frequencies with the largest observed amplification. This conclusion contradicts what has been previously stated from observational and modeling investigations, where the basin itself has been discarded as a preponderant factor promoting long and devastating shaking in Mexico City.


Wheezing in childhood: incidence, longitudinal patterns and factors predicting persistence.
Childhood asthma is frequently perceived as a disease with uniform clinical pathways. This perception might be an oversimplification. The aim of the present study was to investigate the incidence and natural course of wheeze over the first 13 yrs of life and analyse the risk factors predicting wheeze at 11-13 yrs of age. The Multicentre Allergy Study, a German birth cohort, recruited 1,314 children in 1990. Physical examinations, interviews on atopic diseases, immunoglobulin (Ig)E and lung function tests were performed up to 13 yrs of age. Complete data on the course of wheeze were available for 441 children. It was found that incidence of wheezing declined with age. The first wheezing episode was reported by 29, 9 and 9% of participants at < or = 3 (early wheezers), 3-6 (late wheezers), and > 6 yrs (very late wheezers) of age, respectively. Wheezing at the age of 13 yrs was associated with parental atopy, and with IgE sensitisation to common allergens, elevated total IgE and exposure to high levels of indoor allergens in early life. All these associations were remarkably stronger among early wheezers than among early nonwheezers. In conclusion, the relevance of an early expression of atopy as a predictor of wheezing at age 13 yrs declines with increasing age of wheezing onset.


NMR identification of an acyl-adenylate intermediate in the aryl-aldehyde oxidoreductase catalyzed reaction.
A new one-pot synthesis was designed to prepare benzoyl-AMP under anhydrous conditions in N,N-dimethylformamide. Reaction of benzoic acid with N,N'-carbonyldiimidazole and subsequently with 5'-adenosyl monophosphate gave the mixed anhydride in 76% isolated yield. The structure of benzoyl-AMP was confirmed by mass spectroscopy and 1H-, 31P-, and 13C-NMR. The purity of the preparation was greater than 98% as indicated by 31P- and 13C-NMR. Purified aryl-aldehyde oxidoreductase was incubated in NMR tubes together with either carboxy-13C-benzoyl-AMP or carboxy-13C-benzoic acid to demonstrate that benzoyl-AMP is an active intermediate during the enzymatic reduction of benzoic acid to benzaldehyde.


Freezer or non-freezer: clinical assessment of freezing of gait.
Freezing of gait (FOG) is both common and debilitating in patients with Parkinson's disease (PD). Future pathophysiology studies will depend critically upon adequate classification of patients as being either 'freezers' or 'non-freezers'. This classification should be based ideally upon objective confirmation by an experienced observer during clinical assessment. Given the known difficulties to elicit FOG when examining patients, we aimed to investigate which simple clinical test would be the most sensitive to provoke FOG objectively. We examined 50 patients with PD, including 32 off-state freezers (defined as experiencing subjective 'gluing of the feet to the floor'). Assessment including a FOG trajectory (three trials: normal speed, fast speed, and with dual tasking) and several turning variants (180° vs. 360° turns; leftward vs. rightward turns; wide vs. narrow turning; and slow vs. fast turns). Sensitivity of the entire assessment to provoke FOG in subjective freezers was 0.74, specificity was 0.94. The most effective test to provoke FOG was rapid 360° turns in both directions and, if negative, combined with a gait trajectory with dual tasking. Repeated testing improved the diagnostic yield. The least informative tests included wide turns, 180° turns or normal speed full turns. Sensitivity to provoke objective FOG in subjective freezers was 0.65 for the rapid full turns in both directions and 0.63 for the FOG trajectory. The most efficient way to objectively ascertain FOG is asking patients to repeatedly make rapid 360° narrow turns from standstill, on the spot and in both directions.


Genetic disruption of oncogenic Kras sensitizes lung cancer cells to Fas receptor-mediated apoptosis.
Genetic lesions that activate KRAS account for ∼30% of the 1.6 million annual cases of lung cancer. Despite clinical need, KRAS is still undruggable using traditional small-molecule drugs/inhibitors. When oncogenic Kras is suppressed by RNA interference, tumors initially regress but eventually recur and proliferate despite suppression of Kras Here, we show that tumor cells can survive knockout of oncogenic Kras, indicating the existence of Kras-independent survival pathways. Thus, even if clinical KRAS inhibitors were available, resistance would remain an obstacle to treatment. Kras-independent cancer cells exhibit decreased colony formation in vitro but retain the ability to form tumors in mice. Comparing the transcriptomes of oncogenic Kras cells and Kras knockout cells, we identified 603 genes that were specifically up-regulated in Kras knockout cells, including the Fas gene, which encodes a cell surface death receptor involved in physiological regulation of apoptosis. Antibodies recognizing Fas receptor efficiently induced apoptosis of Kras knockout cells but not oncogenic Kras-expressing cells. Increased Fas expression in Kras knockout cells was attributed to decreased association of repressive epigenetic marks at the Fas promoter. Concordant with this observation, treating oncogenic Kras cells with histone deacetylase inhibitor and Fas-activating antibody efficiently induced apoptosis, thus bypassing the need to inhibit Kras. Our results suggest that activation of Fas could be exploited as an Achilles' heel in tumors initiated by oncogenic Kras.


Cholesteryl Ester Storage Disease: An underdiagnosed cause of cirrhosis in adults.
Cholesteryl Ester Storage Disease (CESD), is a rare multisystem autosomal recessive disorder and belongs to the broad family of lysosomal storage disorders. It can present anytime from infancy and childhood to even adulthood. The clinical manifestations are generally severe in infants and with milder forms in adults. One of the prominent sites of involvement is liver. Due to low awareness of this condition among physicians including surgical pathologists, majority of the liver biopsies, especially from the adults are often misdiagnosed as non-alcoholic fatty liver disease/non-alcoholic steatohepatitis or cryptogenic cirrhosis. Given the recent availability of safe and effective enzyme replacement therapy that can alter the natural course of CESD, the pathologists signing out adult and pediatric liver biopsies should be aware of this entity, thus contributing to timely patient management. This review discusses the clinical features, pathogenesis, diagnostic approach, differential diagnosis and management of CESD in adults.


"A lot to manage and still have some kind of a life": How multiple myeloma impacts the function and quality-of-life of Black-White patient-caregiver dyads.
Multiple myeloma (MM) is an incurable debilitating blood cancer associated with the lowest health related quality of life (HRQoL) of all cancers. With nearly 88% of adults aged ≥55 years at diagnosis, age-associated physical losses, comorbidities, and social factors contribute to worsening HRQoL. This qualitative study assessed dyadic (patient-informal caregiver) perspectives on the factors contributing to HRQoL in MM survivors. We recruited 21 dyads from the UNC-Chapel Hill Lineberger Comprehensive Cancer between 11/2021 and 04/2022. Participants completed a single dyadic semistructured interview capturing broad perspectives on MM. We used ATLAS. ti v 9 for project management and to facilitate data analysis using the Sort and Sift, Think and Shift approach (ResearchTalk, Inc.). This iterative approach allowed the exploration and identification of themes within and across transcripts. The mean age at enrollment was 71 years (median: 71, range: 57-90) for patients and 68 years (median 67, range: 37-88) for caregivers. All dyads were racially concordant (11 Black/AA and 10 White). However, we aggregated the findings due to no consistent racial differences. Six themes related to (1) physical burden, (2) treatment challenges, (3) losses of independence, (4) caregiver burden, (5) patient and caregiver perseverance, and (6) adjustment to a new normal were identified. Dyads also experienced MM together, resulting in patients and caregivers experiencing changes in their ability to engage in physical and social activities, which further contributed to poor HRQoL. Patients' increased need for social support led to shifts in the caregiver roles, resulting in caregivers feeling burdened by their responsibilities. All dyads acknowledged the need for perseverance and adaptability to a new normal with MM. The functional, psychosocial, and HRQoL of older patients with MM and their caregivers remain impacted ≥6 months after a new diagnosis highlighting clinical and research opportunities to focus on preserving or improving the health of dyads living with MM.


Transitional cell ovarian carcinoma in a BRCA1 mutation carrier.
BRCA1 mutation carriers are at high risk of developing epithelial ovarian cancer, but the transitional cell variant has not been previously reported in these patients. A nulligravid, perimenopausal woman underwent exploratory laparotomy for a pelvic mass, ascites, and omental caking. Intraoperatively, frozen section of a tumor implant revealed high-grade epithelial ovarian carcinoma. Optimal surgical cytoreduction was performed. The final surgical pathology confirmed International Federation of Gynecology and Obstetrics stage IIIC transitional cell ovarian carcinoma. Her family history was significant for a sister with premenopausal breast cancer and a paternal aunt with ovarian cancer. The patient was counseled and elected to undergo genetic testing. Comprehensive gene sequence analysis detected the germline BRCA1 5382insC mutation. Transitional cell ovarian carcinoma is a rare histologic variant of epithelial ovarian cancer that may occur in BRCA1 mutation carriers.


Quantitative determination of naltrexone and naltrexone prodrugs by electron-capture gas-liquid chromatography.
An electron-capture gas chromatographic method has been applied to the determination of eleven ester and ether derivatives of naltrexone believed to act as prodrugs. Standard analytical curves are presented for all prodrugs and quantitation is shown to be possible from 10 ng to 1.5 microgram of each compound. Ester derivatives of naltrexone are hydrolyzed to naltrexone prior to analysis as the perfluoroalkyl esters. Analysis of synthetic mixtures of these with naltrexone demonstrated that quantitation by difference measurements is possible with naltrexone-derivative ratios from 4:1 to 1:10. Ether derivatives are analyzed without hydrolysis. This method is applicable to biological fluids as well as aqueous solutions.


Seed longevity: survival and maintenance of high germination ability of dry seeds.
The seed constitutes the main vector of plant propagation and it is a critical development stage with many specificities. Seed longevity is a major challenge for the conservation of plant biodiversity and for crop success. Seeds possess a wide range of systems (protection, detoxification, repair) allowing them to survive in the dry state and to preserve a high germination ability. Therefore, the seed system provides an appropriate model to study longevity and aging.


Prostaglandin D2 regulates joint inflammation and destruction in murine collagen-induced arthritis.
Prostaglandin D2 (PGD2) may exert proinflammatory or antiinflammatory effects in different biologic systems. Although this prostanoid and the enzymes responsible for its synthesis are up-regulated by interleukin-1β (IL-1β) in human chondrocytes in vitro, the role of PGD2 in arthritis remains unclear. This study was undertaken to investigate the role of PGD2 in the inflammatory response and in joint destruction during the development of collagen-induced arthritis (CIA) in mice. PGD2 and cytokine levels in mice with CIA were determined by enzyme-linked immunosorbent assay. Expression of hematopoietic PGD synthase (h-PGDS), lipocalin-type PGD synthase (l-PGDS), and DP1 and DP2 receptors was analyzed by immunohistochemical methods. PGE2 levels were determined by radioimmunoassay. The arthritic process up-regulated the expression of h-PGDS, l-PGDS, DP1, and DP2 in articular tissue. PGD2 was produced in the joint during the early phase of arthritis, and serum PGD2 levels increased progressively throughout the arthritic process, reaching a maximum during the late stages of CIA. Treatment of arthritic mice with the DP1 antagonist MK0524 soon after the onset of disease increased the incidence and severity of CIA as well as the local levels of IL-1β, CXCL-1, and PGE2, whereas IL-10 levels were reduced. The administration of the DP2 antagonist CAY10595 did not modify the severity of arthritis. The injection of PGD2 into the paw, as well as the administration of the DP1 agonist BW245C, significantly lowered the incidence of CIA, the inflammatory response, and joint damage. Our findings indicate that PGD2 is produced in articular tissue during the development of CIA and plays an antiinflammatory role, acting through the DP1 receptor.


Investigation of free fatty acid reduction from mixed crude palm oil using 3D-printed rotor-stator hydrodynamic cavitation: An experimental study of geometric characteristics of the inner hole.
A continuous esterification process is employed to decrease the free fatty acid (FFA) concentration of FFA-rich mixed crude palm oil. Both optimal and recommended conditions are determined for the esterification reaction conditions and the geometry of the 3D-printed rotor design in the rotor-stator hydrodynamic cavitation reactor. This study is primarily concerned with the effect of the cavitation device configuration, especially the rotor design, on FFA reduction. Instead of conventional spherical or cylindrical drilled holes, a point angle cone-shaped hole is used to create cavities over the rotor surface. These point angles are adjusted to clarify their effect on FFA reduction. The response surface methodology is applied to determine the optimal concentrations of methanol and sulfuric acid, rotor speed, hole diameter and depth, and cone point angle. The recommended conditions are 20.8 wt% methanol, 2.6 wt% sulfuric acid, 3000 rpm, 5 mm hole diameter, 5 mm hole depth, and 110°, respectively. Under this configuration, the FFA content is reduced from 12.014 wt% to around 1 wt%. A maximum yield of 97.34 vol% esterified oil is obtained through a completed phase separation step, and 93.31 vol% pure oil is collected after the cleansing step. The recommended conditions result in reduced chemical usage, cheaper FFA reduction, and lower environmental impact. This creative rotor design effectively improves our understanding of the geometry of the cavitation device, thus enhancing the cavitation effect in industrial operations.


Health care needs for children of the recently homeless.
Thirty families were surveyed at a shelter for the homeless in San Diego, California concerning the health care status and needs of their children. 56.7% of the families had no regular source of health care and 46.6% were not covered by any form of health coverage. Children whose families had a regular source of health care were more likely to have reported better health and more frequent checkups. Also, increased duration of homelessness was predictably correlated with poorer reported health of the children. Though the families sampled were representative of those recently made homeless i.e. less than six months, needs were identified that would be applicable to all homeless families. The most frequent needs the families expressed for their children were: general nonemergency clinics (76.7%), emergency services (66.7%), and dental services (66.7%). Nutritional and dietary counseling were chosen by 43.3% and only 6.7% of the parents rated social and psychological services as needed.


Pressure ulcers: more lethal than we thought?
To investigate the burden of pressure ulcer-associated mortality in the United States and to examine racial/ethnic differences and associated comorbidities. A descriptive study with matched odds ratio comparisons. The United States, 1990-2001. Pressure ulcer-associated deaths were identified from national multiple cause-coded death records from 1990 to 2001. Age-adjusted mortality rates and matched odds ratio comparisons of pressure ulcer-associated deaths with deaths from other conditions. Between 1990 and 2001, pressure ulcers were reported as a cause of death among 114,380 persons (age-adjusted mortality rate, 3.79 per 100,000 population; 95% confidence interval [CI], 3.77-3.81). For 21,365 (18.7%) of these deaths, pressure ulcers were reported as the underlying cause. Nearly 80% of pressure ulcer-associated deaths occurred in persons at least 75 years old. Septicemia was reported in 39.7% of pressure ulcer-associated deaths (matched odds ratio, 11.3; 95% CI, 11.0-11.7). Multiple sclerosis, paralysis, Alzheimer disease, osteoporosis, and Parkinson disease were reported more often in pressure ulcer-associated deaths than in matched controls. Pressure ulcer-associated mortality was higher among blacks than among whites (age-adjusted rate ratio, 4.22; 95% CI, 4.16-4.27). Pressure ulcers are associated with fatal septic infections and are reported as a cause of thousands of deaths each year in the United States. Incapacitating chronic and neurodegenerative conditions are common comorbidities, and mortality rates in blacks are higher than in other racial/ethnic groups.


Differential toxicity of chronic exposure to phenytoin, phenobarbital, or carbamazepine in cerebral cortical cell cultures.
The effects of phenytoin (30 micrograms/ml), phenobarbital (64 micrograms/ml), and carbamazepine (24 micrograms/ml) were assessed in cerebral cortical cell cultures. After antiepileptic drug exposure for eleven days, cultures were assayed for total protein, number of neurons, tetanus toxin fixation, high-affinity uptake of gamma-aminobutyric acid and beta-alanine, activity of choline acetyltransferase, and benzodiazepine binding. Carbamazepine-exposed cultures demonstrated minimal effects, whereas highly significant deficits related to generalized toxicity were observed in cultures exposed to phenytoin or phenobarbital.


Effects of Leptin, Growth Hormone and Photoperiod on Pituitary SOCS-3 Expression in Sheep.
This study examined how leptin affects growth hormone (GH) release and investigated the effects of leptin, GH, and day length on the suppressor of cytokine signaling-3 (SOCS-3) mRNA levels in the adenohypophyses of sheep. The study consisted of two experiments. The first experiment was conducted during long (LD) and short (SD) days. Within-season and replicate sheep were centrally infused with Ringer-Locke buffer or leptin three times at 60-min intervals at the beginning of experiments. The second experiment involved adenohypophyses collected from sheep that were euthanized in May or November. Pituitary explants were treated with medium alone (Control) or medium with leptin or GH at different concentrations and incubated for various times. The results of the first experiment indicated GH concentrations were seasonally dependent and that leptin had no effect on GH secretion. The results of the second experiment indicated a stronger influence of leptin on the expression of SOCS-3 during the SD season than the LD season. During SDs, significant effects of both GH doses on SOCS-3 expression were observed. These results indicate a strong association between leptin, GH, and SOCS-3, which may explain the disruption of SOCS-3 leptin and GH signaling and the dominant effect of photoperiod on the above relationships.


TGF-beta is required for programmed cell death in interdigital webs of the developing mouse limb.
During limb formation massive cell death in the mesenchyme of the interdigital spaces accompanies the formation of free digits. Members of the transforming growth factor beta (TGF-) superfamily were discussed to play a key role in cell-cell interactions, important in the regulation of programmed cell death (PCD). TGF-beta itself is believed to be involved in epithelial-mesenchymal interactions. Here, we demonstrate that PCD is significantly reduced in interdigital spaces of the developing limbs of Tgfbeta2-/-Tgfbeta3-/- double knockouts. The regression of interdigital webs seems to be doses-dependent as interdigital mesenchyme is at least partly reduced in Tgfbeta2-/-Tgfbeta3+/- mutants, whereas interdigital zones of Tgfbeta2-/-Tgfbeta3-/- double knockouts reveal only minimal signs of regression. We conclude that TGF- is a critical extrinsic regulator of PCD.


Isolation of hepatocytes from postnatal mice.
A method for the isolation of hepatocytes from postnatal (1- to 3-week-old) mice has been developed. Cell isolation was carried out by retrograde perfusion of the liver with a collagenase-containing bicarbonate buffer. Viable cells were separated by selective adsorption onto collagen membranes. Cell viability was assessed by measuring ATP and glutathione content, lactate:pyruvate ratio, and the rate of protein synthesis. Comparisons of these parameters were made with those in cells isolated from adult mice and with values in whole liver. These hepatocytes were capable of metabolizing acetaminophen to its known hepatic conjugates and were susceptible to acetaminophen toxicity. This procedure for isolating mouse hepatocytes should be useful for the study of hepatic drug metabolism and its relationship to toxicity in the postnatal mouse.


Extraordinary phylogenetic diversity and metabolic versatility in aquifer sediment.
Microorganisms in the subsurface represent a substantial but poorly understood component of the Earth's biosphere. Subsurface environments are complex and difficult to characterize; thus, their microbiota have remained as a 'dark matter' of the carbon and other biogeochemical cycles. Here we deeply sequence two sediment-hosted microbial communities from an aquifer adjacent to the Colorado River, CO, USA. No single organism represents more than ~1% of either community. Remarkably, many bacteria and archaea in these communities are novel at the phylum level or belong to phyla lacking a sequenced representative. The dominant organism in deeper sediment, RBG-1, is a member of a new phylum. On the basis of its reconstructed complete genome, RBG-1 is metabolically versatile. Its wide respiration-based repertoire may enable it to respond to the fluctuating redox environment close to the water table. We document extraordinary microbial novelty and the importance of previously unknown lineages in sediment biogeochemical transformations.


Effect of chromatic filters on visual performance in individuals with mild traumatic brain injury (mTBI): A pilot study.
Spectral filters have been used clinically in patients with mild traumatic brain injury (mTBI). However, they have not been formally assessed using objective techniques in this population. Thus, the aim of the present pilot study was to determine the effect of spectral filters on reading performance and visuo-cortical responsivity in adults with mTBI. 12 adults with mTBI/concussion were tested. All reported photosensitivity and reading problems. They were compared to 12 visually-normal, asymptomatic adults. There were several test conditions: three luminance-matched control filters (gray neutral density, blue, and red), the patient-selected 'precision tint lens' that provided the most comfort and clarity of text using the Intuitive Colorimeter System, and baseline without any filters. The Visagraph was used to assess reading eye movements and reading speed objectively with each filter. In addition, both the amplitude and latency of the visual-evoked potential (VEP) were assessed with the same filters. There were few significant group differences in either the reading-related parameters or VEP latency for any of the test filter conditions. Subjective improvements were noted in most with mTBI (11/12). The majority of patients with mTBI chose a tinted filter that resulted in increased visual comfort. While significant findings based on the objective testing were found for some conditions, the subjective results suggest that precision tints should be considered as an adjunctive treatment in patients with mTBI and photosensitivity.


Introducing the Escalation Antibiogram: A Simple Tool to Inform Changes in Empiric Antimicrobials in the Nonresponding Patient.
Hospital antibiograms guide initial empiric antibiotic treatment selections, but do not directly inform escalation of treatment among nonresponding patients. Using gram-negative bacteremia as an exemplar condition, we sought to introduce the concept of an escalation antibiogram. Among episodes of gram-negative bacteremia between 2017 and 2020 from 6 hospitals in the Greater Toronto Area, we generated escalation antibiograms for each of 12 commonly used agents. Among organisms resistant to that antibiotic, we calculated the likelihood of susceptibility to each of the other 11 agents. In subgroup analyses, we examined escalation antibiograms across study years, individual hospitals, community versus hospital onset, and pathogen type. Among 6577 gram-negative bacteremia episodes, the likelihood of coverage was ampicillin 31.8%, cefazolin 62.7%, ceftriaxone 67.1%, piperacillin-tazobactam 72.5%, ceftazidime 74.1%, trimethoprim-sulfamethoxazole 74.4%, ciprofloxacin 77.1%, tobramycin 88.3%, gentamicin 88.8%, ertapenem 91.0%, amikacin 97.5%, and meropenem 98.2%. The escalation antibiograms revealed marked shifts in likelihood of coverage by the remaining 11 agents. For example, among ceftriaxone-resistant isolates, piperacillin-tazobactam susceptibility (21.2%) was significantly lower than trimethoprim-sulfamethoxazole (54.2%, P < .0001), ciprofloxacin (63.0%, P < .0001), ertapenem (73.4%, P < .0001), tobramycin (80.1%, P < .0001), gentamicin (82.8%, P < .0001), meropenem (94.3%, P < .0001), and amikacin (97.1%, P < .0001). Trimethoprim-sulfamethoxazole was the second-ranked agent in the meropenem escalation antibiogram (49.6%) and first in the amikacin escalation antibiogram (86.0%). Escalation antibiograms were consistent across 4 study years and 6 hospitals. Escalation antibiograms can be generated to inform empiric treatment changes in nonresponding patients. These tools can yield important insights such as avoiding the common maneuver of escalating from ceftriaxone to piperacillin-tazobactam in suspected gram-negative bacteremia.


Comprehensive analysis of multiple mycotoxins and Aspergillus flavus metabolites in maize from Kenyan households.
This study assessed the levels of mycotoxins in maize from Kenyan households. Further, local open pollinated maize varieties were compared with commercial hybrids to evaluate which variety is less susceptible to mycotoxin contamination. Four hundred and eighty (n = 480) maize samples were collected in the years 2018-2020 from households in Eastern, Western, Coastal and Lake Victoria regions of Kenya. Liquid chromatography coupled to tandem mass spectrometry was used to detect and quantify 22 mycotoxins, along with 31 Aspergillus flavus metabolites in the samples. Eastern Kenya had the highest aflatoxin (AF) contamination with 75% of samples having AF levels above the Kenyan regulatory limits (10 μg/kg), the highest concentration was 558.1 μg/kg. In Western Kenya, only 18% of samples had concentration levels above the Kenyan regulatory limits for AF with highest sample having 73.3 μg/kg. The Lake Victoria region had the most fumonisins (F) contamination, with 53% of the samples having fumonisin B1 (FB1) < 1000 μg/kg. However, only 20% of the samples surpassed the Kenyan regulatory limit for total fumonisins (2000 μg/kg) with the highest concentration being 13,022 μg/kg. In addition, 21.6% of samples from the Lake Victoria region had zearalenone (ZEN) and deoxynivalenol (DON) above regulatory limits for European countries (1000 μg/kg). Western region had the least A. flavus metabolites contamination (18%) while the Eastern region had the highest incidence of A. flavus metabolites (81%). Among the A. flavus metabolites, cyclopiazonic acid (CPA), beta-cyclopiazonic acid (β CPA), flavacol (FLV) and methylcitreo-isocoumarin (MIC) positively correlated with each other but negatively correlated with the other metabolites. Significant positive co-occurrence was also noted among Fusarium mycotoxins: nivalenol (NIV) positively correlated with DON (r = 0.81), fusarenon-X (FX) (r = 0.81) and ZEN (r = 0.70). Negative correlations were observed between Aspergillus and Fusarium mycotoxins: aflatoxin B1 (AFB1) negatively correlated with FB1 (r = -0.11), FX (r = -0.17) and ZEN (r = -0.20). Local open-pollinated maize varieties (L-opv) were less susceptible to mycotoxin contamination compared to the commercial hybrids (C-hy). This study reveals that Kenyan maize is contaminated with multiple mycotoxins most of which are not regulated in Kenya despite being regulated in other parts of the world. A comprehensive legislation should therefore be put in place to protect the Kenyan public against chronic exposure to these mycotoxins. In addition to high yield, there is a need for commercial hybrid maize breeders to incorporate mycotoxin resistance as an important trait in germplasm improvement in seeds production.


A novel method for producing unequal sized droplets in micro- and nanofluidic channels.
We propose a novel method for producing unequal sized droplets through breakup of droplets. This method does not have the disadvantages of the available methods and also reduces the dependence of the droplets volume ratio on the inlet velocity of the system by up to 26 percent. The employed method for investigating the proposed system relies on 3D numerical simulation using the VOF algorithm and the results have been obtained with various valve ratios for both the micro- and nanoscale. The results indicate that the droplet length during the breakup process increases linearly with time. The droplet length at the nanoscale is smaller than that at the micro scale. It has been shown that the maximum local capillary number in this system is 2.5 times the average capillary number. Therefore one can use the analytical theories based on the low capillary number assumptions to investigate the method.


Target Lesion Failure With Current Drug-Eluting Stents: Evidence From a Comprehensive Network Meta-Analysis.
The aim of this study was to investigate the efficacy and safety of currently used drug-eluting stents (DES). Head-to-head comparisons among newer DES have shown conflicting results. For this network meta-analysis, randomized controlled trials comparing different types of currently used DES were searched in PubMed, Scopus, and proceedings of international meetings. The primary endpoint was target lesion failure (TLF) at 1 year and at long-term follow-up. Seventy-seven trials with 99,039 patients were selected for this network meta-analysis. Among the 10 DES included in the meta-analysis, 4 received the most extensive investigation: Orsiro, XIENCE, Nobori/BioMatrix, and Resolute. At 1 year, the Orsiro stent was associated with lower rates of TLF compared with XIENCE (odds ratio [OR]: 0.84; 95% confidence interval [CI]: 0.71 to 0.98; p = 0.03), Resolute (OR: 0.81; 95% CI: 0.68 to 0.95; p = 0.01), and Nobori/BioMatrix (OR: 0.81; 95% CI: 0.67 to 0.98; p = 0.03). Orsiro had the highest probability to be the best (70.8%), with a surface under the cumulative ranking curve value of 95.9%. However, after a median follow-up period of 50 months (range: 24 to 60 months), no significant difference was apparent in the rates of TLF between any DES, although Orsiro still ranked as the best stent (58.6% probability to be the best). In addition, Orsiro had a lower rate of long-term definite stent thrombosis compared with Nobori/BioMatrix (OR: 0.60; 95% CI: 0.36 to 0.98; p = 0.04) and lower rates of definite and probable stent thrombosis compared with Resolute (OR: 0.66; 95% CI: 0.45 to 0.99; p = 0.04). No differences in cardiac mortality between any DES were observed. Orsiro is associated with a lower 1-year rate of TLF compared with XIENCE, Resolute, and Nobori/BioMatrix but with an attenuation of the efficacy signal at long-term follow-up.


Topological information embodied in local juxtaposition geometry provides a statistical mechanical basis for unknotting by type-2 DNA topoisomerases.
Topoisomerases may unknot by recognizing specific DNA juxtapositions. The physical basis of this hypothesis is investigated by considering single-loop conformations in a coarse-grained polymer model. We determine the statistical relationship between the local geometry of a juxtaposition of two chain segments and whether the loop is knotted globally, and ascertain how the knot/unknot topology is altered by a topoisomerase-like segment passage at the juxtaposition. Segment passages at a "free" juxtaposition tend to increase knot probability. In contrast, segment passages at a "hooked" juxtaposition cause more transitions from knot to unknot than vice versa, resulting in a steady-state knot probability far lower than that at topological equilibrium. The reduction in knot population by passing chain segments through a hooked juxtaposition is more prominent for loops of smaller sizes, n, but remains significant even for larger loops: steady-state knot probability is only approximately 2%, and approximately 5% of equilibrium, respectively, for n=100 and 500 in the model. An exhaustive analysis of approximately 6000 different juxtaposition geometries indicates that the ability of a segment passage to unknot correlates strongly with the juxtaposition's "hookedness". Remarkably, and consistent with experiments on type-2 topoisomerases from different organisms, the unknotting potential of a juxtaposition geometry in our polymer model correlates almost perfectly with its corresponding decatenation potential. These quantitative findings suggest that it is possible for topoisomerases to disentangle by acting selectively on juxtapositions with "hooked" geometries.


Nanostring technology on Fibrous Dysplasia bone biopsies. A pilot study suggesting different histology-related molecular profiles.
Identifying the molecular networks that underlie Fibrous Dysplasia (FD) is key to understand the pathogenesis of the disease, to refine current diagnostic approaches and to develop efficacious therapies. In this study, we used the NanoString nCounter Analysis System to investigate the gene signature of a series of nine Formalin Fixed Decalcified and Paraffin-Embedded (FFDPE) bone biopsies from seven FD patients. We analyzed the expression level of 770 genes. Unsupervised clustering analysis demonstrated partitioning into two clusters with distinct patterns of gene expression. Differentially expressed genes included growth factors, components of the Wnt signaling system, interleukins and some of their cognate receptors, ephrin ligands, matrix metalloproteinases, neurotrophins and genes encoding components of the cAMP-dependent protein kinase. Interestingly, two tissue samples obtained from the same skeletal site of one patient one year apart failed to segregate in the same cluster. Retrospective histological review of the samples revealed different microscopic aspects in the two groups. The results of our pilot study suggest that the genetic signature of FD is heterogeneous and varies according to the histology and, likely, to the age of the lesion. In addition, they show that the Nanostring technology is a valuable tool for molecular translational studies on archival FFDPE material in FD and other rare bone diseases.


40Hz auditory steady-state responses in patients with disorders of consciousness: Correlation between phase-locking index and Coma Recovery Scale-Revised score.
We aimed to elucidate whether 40Hz auditory steady-state response (ASSR) could be sensitive to the state of patients with disorders of consciousness (DOC) as estimated with Coma Recovery Scale-Revised (CRS-R) diagnostic tool. Fifteen DOC patients and 24 healthy controls took part in the study. The 40Hz click trains were used to evoke ASSRs. Mean evoked amplitude (EA) and phase-locking index (PLI) within 38-42Hz window were calculated for 100ms bins, starting from -200 to 700ms relative to stimulus onset. The PLI values from the patient group in the period of 200-500ms after the stimulus onset positively correlated with the CRS-R total score and with the scores of the Auditory and Visual subscales. The phase-locking index of 40Hz auditory steady-state responses can be an indicator of the level of dysfunction of the central nervous system in DOC. Our results emphasize the role of central auditory system integrity in determining the level of functioning of DOC patients and suggest the possibility to use the ASSR protocol as an objective diagnostic method in DOC patients.


Spatial modeling of zoonotic cutaneous leishmaniasis with regard to potential environmental factors using ANFIS and PCA-ANFIS methods.
This study compares two adaptive neuro-fuzzy inference system (ANFIS) and principal component analysis (PCA)-ANFIS techniques for spatial modeling and forecasting of zoonotic cutaneous leishmaniasis (ZCL) cases in rural districts of Golestan province, Iran. We collected and prepared data on ZCL cases and climatic, topographic, vegetation, and human population factors. By applying the PCA algorithm, the parameters affecting the ZCL incidence were decomposed into principal components (PCs), and their dimensions were reduced. Then, PCs were used to train the ANFIS model. To evaluate the proposed approaches in model assessment phase, we used test data in 2016. In this phase, we showed that PCA-ANFIS model with values of R2 = 0.791, MAE = 0.681, RMSE = 0.904 compared to ANFIS model with values of R2 = 0.705, MAE = 0.827, RMSE = 1.073 has better performance in prediction of the ZCL cases. Actual and predicted maps of ZCL cases in 2016 by both models demonstrated that the high-risk regions of the disease are located in the northeastern, northern parts, and some central rural districts of Golestan province. Sensitivity analysis of the ANFIS model showed that population, vegetation, average wind speed, elevation, and average soil temperature, respectively, are the most significant factors in predicting the ZCL cases. The findings indicated the importance of machine learning (ML) techniques (ANFIS and PCA-ANFIS) in medical geography studies. By using these approaches, with less cost and shorter time, high-risk areas of diseases can be predicted, and the most effective factors on the spatial prediction of diseases can be identified. Public health policymakers can use these useful tools to control and prevent the disease and to allocate resources to disease-prone areas.


Stress and glucocorticoids increase transthyretin expression in rat choroid plexus via mineralocorticoid and glucocorticoid receptors.
Transthyretin (TTR) is a carrier for thyroid hormones and retinol binding protein. Several mutated forms of TTR cause familial amyloidotic polyneuropathy, an inheritable lethal disease. On the other hand, wild-type TTR has a protective role against Alzheimer's disease. Despite its overall importance in normal animal physiology and in disease, few studies have focused on its regulation. An in silico analysis of the rat TTR gene revealed a glucocorticoid responsive element in the 3' region of the first intron. Thus, we hypothesised that TTR could be regulated by glucocorticoid hormones and investigated the regulation of TTR expression in response to hydrocortisone in a rat choroid plexus cell line (RCP) and in primary cultures of choroid plexus epithelial cells (CPEC). In addition, the effect of psychosocial stress on TTR expression was analysed in rat liver, choroid plexus (CP) and cerebrospinal fluid (CSF). In RCP and CPEC cultures hydrocortisone upregulated TTR expression, an effect suppressed by glucocorticoid receptor and mineralocorticoid receptor antagonists. Moreover, induction of psychosocial stress increased TTR expression in liver, CP and CSF of animals subjected to acute and chronic stress conditions. Overall, we conclude that stress upregulates TTR expression in CP.


Slide tracheoplasty: a case report of successful concomitant reconstruction of extensive congenital tracheal stenosis and pulmonary artery sling.
An 8-month-old infant presented with an extensive congenital tracheal stenosis with an aberrant left pulmonary artery. The patient was treated successfully by relocation of the left pulmonary artery and tracheal reconstruction with slide tracheoplasty. This patient is the first survivor cited in the literature after concomitant repair using slide tracheoplasty.


[Use of recombinant factor VIIa to reduce transfusional load in hemorrhaging patients after surgery for malignant tumors].
Often expanded surgical interventions in patients with malignant tumors are accompanied by huge bleeding and require transfusion of big mass of medications prepared from donor blood that may develop different complications. It is showed that with the aim of treatment of hemorrhagic complications the use of recombination factor VIIa allows reducing the surgical blood loss, as well as diminishing hemotransfusional load after surgery especially in patients with malignancies of urogenital system and malignant tumors of the colon and lung.


Different Modes of Acid-Promoted Cyclooligomerization of 4-(4-Thiosemicarbazido)butan-2-one Hydrazone: 14-Membered versus 28-Membered Polyazamacrocycle Formation.
Unprecedented self-assembly of a novel 14-membered cyclic bis-thiosemicarbazone or/and a 28-membered cyclic tetrakis-thiosemicarbazone upon acid-promoted cyclooligomerization of 4-(4-thiosemicarbazido)butan-2-one hydrazone has been discovered. A thorough study of the influence of various factors on the direction of macrocyclization provided the optimal conditions for the highly selective formation of each of the macrocycles in excellent yields. Plausible pathways for macrocyclizations have been discussed. The macrocycle precursor was prepared by the reaction of readily available 4-isothiocyanatobutan-2-one with an excess of hydrazine.


The novel isoflavone derivatives inhibit RANKL-induced osteoclast formation.
Isoflavones are compounds structurally similar to the mammalian estrogens and have received considerable attention for their preventive actions on bone loss. Here, we synthesized the novel isoflavone derivatives and examined their activities in bone cells. We found that the novel isoflavone derivatives markedly inhibited the receptor activator of nuclear factor kappa B ligand (RANKL) plus macrophage colony stimulating factor (M-CSF)-induced osteoclastic differentiation from bone marrow stromal cells and RAW264.7 macrophage cells. Treatment of RAW264.7 macrophages with RANKL-induced extracellular signal-regulated kinase (ERK), p38 and c-Jun N-terminal kinase (JNK) phosphorylation. However, RANKL-induced p38 and JNK but not ERK phosphorylation was attenuated by isoflavone derivatives. Furthermore, RANKL-mediated increase of p65 phosphorylation at Ser⁵³⁶, NF-κB-specific DNA-protein complex formation and κB-luciferase activity was inhibited by isoflavone derivatives. On the other hand, isoflavone derivatives did not affect the cell proliferation and differentiation of human cultured osteoblasts. Our data suggest that the novel isoflavone derivatives inhibit osteoclastogenesis from bone marrow stromal cells and macrophage cells via attenuation of RANKL-induced p38, JNK and NF-κB activation.


Tumor budding to investigate local invasion, metastasis and prognosis in temporal bone squamous cell carcinoma.
Few studies have tried to go beyond the conventional clinic-pathological prognostic factors investigating the molecular markers involved in the biology of temporal bone squamous cell carcinoma (TBSSC). Tumor budding represents a very aggressive subpopulation of cancer cells and demonstrates the heterogeneity of cancer cells residing in different locations within tumors. The main aim of this exploratory study was to investigate the role of tumor budding in primary TBSCC prognosis. We also analyzed the association between TBSCC tumor budding and: (i) loco-regional aggressiveness evaluated according to the revised Pittsburgh staging system, (ii) tumor infiltrating lymphocytes, lymphovascular invasion (LVSI), perineural invasion, pattern of invasion, and type of stroma. Thirty-two TBSCCs treated surgically were considered. The three-tier grading system recommended by the International Tumor Budding Consensus Conference was used first on TBSCC. Advanced (T3-4) TBSCC was related with high risk intra-tumoral budding (ITB) at two-tier risk grading (p = 0.0361). N + status was associated with intermediate/high budding (Bd2-3) at three-tier risk grading for peri-tumoral budding (PTB) (p = 0.0382). Disease-free survival (DFS) was related with T-stage (p = 0.0406), N-status (p < 0.0001), PTB two-tier risk grading (p = 0.0463), LVSI (p < 0.0001). Overall survival (OS) was associated with N-status (p = 0.0167), PTB absolute count (p = 0.0341), PTB three-tier risk grading (p = 0.0359), PTB two-tier risk grading (p = 0.0132), and LVSI (p = 0.0004). At the multivariate analysis, DFS was related with N-status (p = 0.0147) and LVSI (p < 0.0001), while OS resulted associated only with LVSI (p = 0.0144). Our preliminary findings suggest that tumor budding in TBSCC, regardless of its localization (the main tumor body [ITB] or invasive front [PTB]) may be a reliable predictor of neck lymph node metastasis and poor prognosis. Tumor budding and LVI could be predictive markers for precise treatment in TBSCC. Further investigations on larger prospective series should be designed to confirm this evidence both in post-operative specimens and in preoperative biopsies.


Prospective cohort study investigating quality of life outcomes following multi-speciality robotic-assisted surgery.
Despite recent evidence on the surgical outcomes of robotic-assisted surgery (RAS), other patient centre outcomes, including quality of life (QOL), are lacking. This study aims to examine changes in QoL trajectories following RAS across different surgical specialities. A prospective cohort study was conducted for patients undergoing urologic, cardiothoracic, colorectal or benign gynaecological RAS, between June 2016 and January 2020 at a tertiary referral hospital in Australia. QoL was measured using the 36-item Short-Form Health Survey at pre-operative, 6 weeks and 6 months postoperatively. Physical and mental summary scores and utility index were primary outcomes, and sub-domains were secondary outcomes. Mixed-effects linear regressions were used to determine changes in QoL trajectories. Of the 254 patients undergoing RAS, 154 underwent urologic, 36 cardiothoracic, 24 colorectal and 40 benign gynaecological surgery. Overall, the average age was 58.8 years and most patients were male (75.1%). Physical summary scores significantly decreased from pre-operative to 6 weeks' post-operative in urologic and colorectal RAS; with all surgical specialities at least returning to pre-operative levels within 6 months postoperatively. Mental summary scores consistently increased from pre-operative to 6 months postoperatively for colorectal and gynaecological RAS. RAS contributed to positive changes in QoL, with physical health returning to the pre-operative level and mental health improvements across specialities, in the short term. While degrees of post-operative changes varied amongst specialities, significant improvements demonstrate benefits in RAS.


Local delivery of interleukin-2 and adriamycin is synergistic in the treatment of experimental malignant glioma.
Local delivery of adriamycin (ADR) via biodegradable polymers has been shown to improve survival in rats challenged intracranially with 9L gliosarcoma. Likewise, local delivery of interleukin-2 (IL-2) has been shown to extend survival in experimental brain tumor models. In the current study, we hypothesized that local delivery of ADR and IL-2 might act synergistically against experimental intracranial glioma. Polyanhydride polymers (PCPP-SA) containing 5% ADR by weight were prepared using the mix-melt method. IL-2 polymer microspheres (IL-2 MS) were produced via the complex coacervation of gelatin and chondroitin sulfate in the presence of IL-2. Sixty male Fisher 344 rats received an intracranial challenge with a lethal dose of 9L gliosarcoma cells. In addition, a group of rats were injected with either IL-2 MS or empty microspheres. Five days later they received ADR or blank polymer. There were a total of four treatment groups: (1) empty microspheres, blank polymer; (2) empty microspheres, ADR polymer; (3) IL-2 MS, blank polymer; and (4) IL-2 MS, ADR polymer. Compared to control animals treated with empty microspheres and blank polymer, animals receiving empty microspheres and ADR polymer (P < 0.0004), IL-2 MS and blank polymer (P < 0.0005), and IL-2 MS combined with ADR polymer (P < 0.0000002) all showed statistically significant improvement in survival. In addition, animals receiving the IL-2/ADR combination had significantly extended survival compared to either ADR or IL-2 alone (P < 0.000003 and P < 0.0004, respectively). Both ADR and IL-2, when delivered locally, are effective monotherapeutic agents against experimental intracranial gliosarcoma. The combination ADR and IL-2 therapy is more effective than either agent alone.


Extracapsular dissection in peripheral nerve schwannoma surgery using bright light and fluorescein sodium visualization: case series.
Schwannomas are the most frequent peripheral nerve sheath tumors and are treated by surgical resection when symptomatic. Tumor removal is performed by intraneural dissection and enucleation. In order to safely remove the tumor from the nerve, the use of sodium fluorescein has recently been proposed to distinguish the tumor from the adjacent normal nerve fibers, before incision of the tumor pseudocapsule and during intraneural tumor dissection. We report a consecutive case series of 5 peripheral nerve schwannomas operated in 4 patients, in which we evaluate the usefulness of sodium fluorescein compared to usual visual landmarks, at each step of the surgical procedure. After exposition of the schwannoma, sodium fluorescein helped with the localization of intracapsular en passant nerve fascicles in only one case. Hence, the definition of a safe entry zone for capsular incision relied mainly on nerve monitoring and direct visualization of en passant nerve fascicles under microscope. During intraneural dissection, there was a sharp contrast between the fluorescent tumor and the non-fluorescent adjacent pseudocapsule in most cases but the colorimetric variation between tumor and normal tissue induced by fluorescence did not outperform the natural contrast between the yellow true capsule and the gray-red layers of the pseudocapsule. Based on these results, we consider that the limited additional value of sodium fluorescein in primary peripheral nerve schwannoma surgery does not warrant its use in daily clinical practice. Additional studies are needed to assess its usefulness during the surgery of recurrences and tumors which are intertwined with several fascicles of origin such as neurofibromas.


Program practices and demographic factors associated with federal funding for the Safe Routes to School program in the United States.
In 2005, the United States Congress authorized $612 million for use in implementing the US Safe Routes to School program to address physical inactivity, air quality, safety and traffic near schools. Each US state developed administrative practices to implement the program. Based on state-specific annual obligations, on average, states have obligated 44% of available funds. State project obligations were directly associated with programmatic factors, including broader adherence to federal agency administrative guidance objectives and the number of years for which the states obligated new projects and indirectly associated with student enrollment and state child poverty. Research and policy recommendations are discussed.


Patterns of utilization and cost of healthcare services and pharmacotherapy among patients with drug-resistant epilepsy during the two-year period before neurostimulation: A descriptive analysis of the journey to implantation based on analyses of a large United States healthcare claims database.
To conduct a descriptive assessment of patterns of utilization and cost of healthcare services and pharmacotherapies among patients with drug-resistant epilepsy (DRE) before neurostimulator implantation. Using a large United States healthcare claims database, we identified all patients with DRE who were implanted with neurostimulators between January 1, 2012, and December 31, 2019. Patients without an epilepsy diagnosis on their implantation date were excluded, as were those without (1) anti-seizure medication (ASM) dispenses within 12 months of implantation date, and (2) continuous enrollment for the 24-month period before this date. Demographic and clinical characteristics were assessed over the two-year period before implantation, as were patterns of utilization and cost of healthcare services and pharmacotherapy. Care was assessed as all-cause or epilepsy-related, with the latter defined as all medical (inpatient and outpatient) care resulting in diagnoses of epilepsy and all ASM dispenses. Eight hundred sixty patients met all selection criteria. Among these patients, comorbidities were common, including depression (27%), anxiety (30%), and learning disabilities (25%). Fifty-nine percent of patients had ≥1 all-cause hospitalizations; 57% had ≥1 epilepsy-related admissions. Patients averaged 8.6 epilepsy-related visits to physicians' offices, including 5.1 neurologist visits. Mean all-cause and epilepsy-related healthcare costs during the pre-implantation period were $123,500 and $91,995, respectively; corresponding median values were $74,567 and $53,029. Median monthly all-cause healthcare costs increased by 138% during the 24-month period (from $1,042 to $2,481 in the month prior to implantation); median epilepsy-related costs, by 290% (from $383 to $1,492). The two-year period before neurostimulator implantation is a long and costly journey. Estimates likely minimize the burden experienced during this period, given that seizure frequency and severity-and corresponding impacts on quality of life-were unavailable in these data. Further research is needed to understand the clinical, economic, and psychological impact of the time between DRE onset and implantation among qualifying patients.


Liquid-solid extraction coupled with magnetic solid-phase extraction for determination of pyrethroid residues in vegetable samples by ultra fast liquid chromatography.
In this study, liquid-solid extraction coupled with magnetic solid-phase extraction was successfully developed for the extraction of pyrethroid residues in vegetable samples. The analytes were determined by ultra fast liquid chromatography. The pyrethroids were extracted by liquid-solid extraction and then adsorbed onto magnetic adsorbent. Magnetic adsorbent, C18-functionalized ultrafine magnetic silica nanoparticles, was synthesized by chemical coprecipitation, silanization and alkylation. The analytes adsorbed onto the magnetic adsorbent can be simply and rapidly isolated from sample solution with a strong magnet on the bottom of the extraction vessel. The extraction parameters, such as liquid-solid extraction solvent, liquid-solid extraction time, the amount of magnetic adsorbent, magnetic solid-phase extraction time and magnetic solid-phase extraction desorption solvent, were optimized to improve the extraction efficiency. The analytical performances of this method, including linear range, detection limit, precision, and recovery were evaluated. The limits of detection for pyrethroid were between 0.63 and 1.2 ng g(-1). Recoveries obtained by analyzing the four spiked vegetable samples were between 76.0% and 99.5%. The results showed that the present method was a simple, accurate and high efficient approach for the determination of pyrethroids in the vegetable samples.


Canadian evidence-based guideline for treatment of relapsed/refractory chronic lymphocytic leukemia.
Following the recent publication of Canadian evidence-based guidelines for frontline treatment of chronic lymphocytic leukemia (CLL), the same group of clinicians developed guidelines for CLL in the relapsed/refractory (R/R) setting. The treatment of R/R CLL has changed significantly in the past few years, with many novel therapeutics available to hematologists across the country. These guidelines aim to standardize the management of CLL in the relapsed/refractory setting, using the best evidence currently available.


Complete genome sequence of a new botybirnavirus isolated from a phytopathogenic Alternaria alternata in China.
A new double-stranded RNA (dsRNA) virus named Alternaria alternata botybirnavirus 1 (AaBRV1) was isolated from Alternaria alternata strain SD-BZF-19, a phytopathogenic fungus infecting watermelon in China. The genome of AaBRV1 consists of two dsRNA segments (dsRNAs 1 and 2), 6,130 and 5,862 bp in size, respectively. The sequence contains two putative open reading frames (ORFs) which encode two polyproteins of 1,874 and 1,784 amino acids, respectively. Nucleotide sequence comparisons revealed that the two ORFs of AaBRV1 have the highest similarity 60.3% and 56.7%, respectively, with dsRNAs 1 and 2 of Botrytis porri RNA virus 1 (BpRV1). The two polyproteins encoded by dsRNA1 and dsRNA2 shared the highest amino acid identities with the cap-pol fusion protein (60.2%) and hypothetical protein (53.7%) of BpRV1, respectively. AaBRV1 is composed of isometric particles, approximately 35 nm in diameter. Phylogenetic analysis of the RNA dependent RNA polymerase (RdRp) domain of the polyprotein revealed that AaBRV1 clusters together with members of the genus Botybirnavirus. These findings support the discovery of a new botybirnavirus in A. alternata.


[Subjective impairment after macroamputation of the upper extremity].
The subjective outcome following macroamputation of the upper limb was evaluated using the DASH score. In a retrospective study, 11 patients with stumps of the upper limb following trauma were investigated 5.6 years after injury. Data according to injury pattern, age, gender, level of amputation and DASH score were collected. Included are 2 unilateral and 3 bilateral forearm amputees and 6 patients with upper arm amputation. Only males were seen with a mean age of 32 years. Amputation occurred following polytrauma in 3 cases while in the other 8 cases the amputated limb was unreconstructably mangled. 8 patients wear a prosthesis. 4 (36 %) patients are unemployed. The mean DASH score was 44.8, in bilateral forearm amputees 74.16, in unilateral upper-arm amputees 38.47 and in unilateral lower forearm amputees 43.33. Patients with bilateral amputation showed the greatest functional and social impairment. Mean pain was 40 % of the pain section in the DASH questionnaire and showed no difference with regard to the level of amputation or bilateral injury. All patients are socially integrated. Patients with unilateral amputation rated psychological impairment higher than functional impairment. Patients with macroamputation of the upper extremity show high psychological and functional impairment. Pain, deficits in function and still not optimal prosthetic devices result in a high percentage of unemployment.


Green synthesis of biogenic silver nanomaterials using Raphanus sativus extract, effects of stabilizers on the morphology, and their antimicrobial activities.
The present study explores the reducing and capping potentials of aqueous Raphanus sativus root extract for the synthesis of silver nanomaterials for the first time in the absence and presence of two stabilizers, namely, water-soluble starch and cetyltrimethylammonium bromide (CTAB). The surface properties of silver nanoparticles (AgNPs) were determined by dynamic light scattering (DLS), transmission electron microscopy (TEM), energy dispersion X-ray spectroscopy (EDX), scanning electron microscopy (SEM), X-ray diffraction (XRD), and Fourier transform infrared (FT-IR) techniques. The mean size of AgNPs, ranging from 3.2 to 6.0 nm, could be facilely controlled by merely varying the initial [extract], [CTAB], [starch], and [Ag(+)] ions. The agglomeration number, average number of silver atoms per nanoparticle, and changes in the fermi potentials were calculated and discussed. The AgNPs were evaluated for their antimicrobial activities against different pathogenic organisms. The inhibition action was due to the structural changes in the protein cell wall.


Diffusion-weighted MR imaging of normal and abnormal scrotum: preliminary results.
Magnetic resonance (MR) imaging of the scrotum represents an important supplemental diagnostic tool in the evaluation of scrotal diseases. Diffusion-weighted (DW) MR imaging is a developing technique, proved to improve tissue characterization. We evaluated the feasibility and diagnostic performance of DW MR imaging in the detection and characterization of scrotal lesions. We retrospectively evaluated 31 scrotal lesions (23 intratesticular and 8 extratesticular) in 26 men. All MR examinations were performed on a 1.5-T unit, using a pelvic-phased array coil. DW sequences were obtained using a single shot, multislice spin echo planar diffusion pulse sequence and a b factor of 0 and 900 s mm(-2). The DW MR characteristics and the apparent diffusion coefficient (ADC) values of normal scrotal contents and scrotal diseases were evaluated. Comparison between the ADC values of normal scrotum, benign lesions and scrotal malignancies was performed. The accuracy of conventional sequences, DW images alone and DW imaging combined with conventional images in differentiating benign from malignant scrotal lesions was calculated. The ADC values of testicular malignancies were different from those of normal testis and benign intratesticular lesions, and the ADC values of benign extratesticular lesions from those of normal epididymis (P<0.05). The overall accuracy of conventional imaging, DW imaging alone and DW MR combined with conventional sequences in the characterization of intratesticular lesions was 91%, 87% and 100%, respectively. Our findings suggest that DW MR imaging and ADC values may provide valuable information in the diagnosis and characterization of scrotal diseases.


Cross-reactions in the HLA system revealed by red blood cells expressing HLA determinants, with particular reference to cross-reaction between HLA-A2 and B17.
Sera with cytotoxic and haemagglutinating activity towards HLA-A2/28 were also shown to react with red blood cells (RBC) expressing the HLA-B17 antigen determinant. Absorption procedures with white blood cells (WBC) indicated that this was due to an HLA-A2/B17 cross-reaction. Absorption experiments with some other sera which previously had shown cytotoxic and haemagglutinating activity towards different HLA specificities, revealed broad cross-reaction related to HLA-B locus antigens.


Effect of Insulin Glargine Up-titration vs Insulin Degludec/Liraglutide on Glycated Hemoglobin Levels in Patients With Uncontrolled Type 2 Diabetes: The DUAL V Randomized Clinical Trial.
Achieving glycemic control remains a challenge for patients with type 2 diabetes, even with insulin therapy. To assess whether a fixed ratio of insulin degludec/liraglutide was noninferior to continued titration of insulin glargine in patients with uncontrolled type 2 diabetes treated with insulin glargine and metformin. Phase 3, multinational, multicenter, 26-week, randomized, open-label, 2-group, treat-to-target trial conducted at 75 centers in 10 countries from September 2013 to November 2014 among 557 patients with uncontrolled diabetes treated with glargine (20-50 U) and metformin (≥1500 mg/d) with glycated hemoglobin (HbA1c) levels of 7% to 10% and a body mass index of 40 or lower. 1:1 randomization to degludec/liraglutide (n = 278; maximum dose, 50 U of degludec/1.8 mg of liraglutide) or glargine (n = 279; no maximum dose), with twice-weekly titration to a glucose target of 72 to 90 mg/dL. Primary outcome measure was change in HbA1c level after 26 weeks, with a noninferiority margin of 0.3% (upper bound of 95% CI, <0.3%). If noninferiority of degludec/liraglutide was achieved, secondary end points were tested for statistical superiority and included change in HbA1c level, change in body weight, and rate of confirmed hypoglycemic episodes. Among 557 randomized patients (mean: age, 58.8 years; women, 49.7%), 92.5% of patients completed the trial and provided data at 26 weeks. Baseline HbA1c level was 8.4% for the degludec/liraglutide group and 8.2% for the glargine group. HbA1c level reduction was greater with degludec/liraglutide vs glargine (-1.81% for the degludec/liraglutide group vs -1.13% for the glargine group; estimated treatment difference [ETD], -0.59% [95% CI, -0.74% to -0.45%]), meeting criteria for noninferiority (P < .001), and also meeting criteria for statistical superiority (P < .001). Treatment with degludec/liraglutide was also associated with weight loss compared with weight gain with glargine (-1.4 kg for degludec/liraglutide vs 1.8 kg for glargine; ETD, -3.20 kg [95% CI, -3.77 to -2.64],P < .001) and fewer confirmed hypoglycemic episodes (episodes/patient-year exposure, 2.23 for degludec/liraglutide vs 5.05 for glargine; estimated rate ratio, 0.43 [95% CI, 0.30 to 0.61],P < .001). Overall and serious adverse event rates were similar in the 2 groups, except for more nonserious gastrointestinal adverse events reported with degludec/liraglutide (adverse events, 79 for degludec/liraglutide vs 18 for glargine). Among patients with uncontrolled type 2 diabetes taking glargine and metformin, treatment with degludec/liraglutide compared with up-titration of glargine resulted in noninferior HbA1c levels, with secondary analyses indicating greater HbA1c level reduction after 26 weeks of treatment. Further studies are needed to assess longer-term efficacy and safety. clinicaltrials.gov Identifier: NCT01952145.


Tracking pathophysiological processes in Alzheimer's disease: an updated hypothetical model of dynamic biomarkers.
In 2010, we put forward a hypothetical model of the major biomarkers of Alzheimer's disease (AD). The model was received with interest because we described the temporal evolution of AD biomarkers in relation to each other and to the onset and progression of clinical symptoms. Since then, evidence has accumulated that supports the major assumptions of this model. Evidence has also appeared that challenges some of our assumptions, which has allowed us to modify our original model. Refinements to our model include indexing of individuals by time rather than clinical symptom severity; incorporation of interindividual variability in cognitive impairment associated with progression of AD pathophysiology; modifications of the specific temporal ordering of some biomarkers; and recognition that the two major proteinopathies underlying AD biomarker changes, amyloid β (Aβ) and tau, might be initiated independently in sporadic AD, in which we hypothesise that an incident Aβ pathophysiology can accelerate antecedent limbic and brainstem tauopathy.


Whether high government subsidies reduce the healthcare provision of township healthcare centers in rural China.
China's government launched a large-scale healthcare reform from 2009. One of the main targets of this round reform was to improve the primary health care system. Major reforms for primary healthcare institutions include increasing government investment. However, there are insufficient empirical studies based on large sample to catch long-term effect of increased government subsidy and lack of sufficient incentives on township healthcare centers (THCs), therefore, this study aims to provide additional empirical evidence on the concern by conducting an empirical analysis of THCs in Shaanxi province in China. We collected nine years (2009 to 2017) data of THCs from the Health Finance Annual Report System (HFARS) that was acquired from the Health Commission of Shaanxi Province. We applied two-way fixed effect model and continue difference-in-difference (DID) model to estimate the effect of percentage of government subsidy on medical provision. A clear jump of the average percentage of government subsidy to total revenue of THCs can be found in Shaanxi province in 2011, and the average percentage has been more than 60% after 2011. Continue DID models indicate every 1% percentage of government subsidy to total revenue increase after 2011 resulted in a decrease of 1.1 to 3.5% in THCs healthcare provision (1.9% in medical revenue, 1.2% in outpatient visit, 3.5% in total occupy beds of inpatient, 1.1% in surgery revenue, 2.1% in sickbed utilization rate). The results show that the THCs with high government subsidy reduce the number of medical services after 2011. We think that it is no doubt that the government should take more responsibility for the financing of primary healthcare institutions, the problem is when government plays a central role in the financing and delivery of primary health care services, more effective incentives should be developed.


A method of measuring pupil-blocking force in the human eye.
A new method of measuring pupil-blocking force in the human eye is reported. The anterior segment of the eye was photographed using a slit-lamp apparatus and the color photographs then analyzed with a digitizer for converting the optical image of the anterior eye segment into the true image using a computer. This technique permits precise estimation of the relative positions of the anterior chamber, such as pupil margins, iris root insertions, and central anteroposterior pupillary axis. Pupil-blocking force was calculated from these data according to Hook's law. There was a statistically significant difference in pupil-blocking force between normal subjects (mean +/- SD, -0.027 +/- 0.029 lambda; lambda = Young's module) and patients with primary angle-closure glaucoma (mean +/- SD, +0.134 +/- 0.068 lambda) (P less than 0.001). In patients with primary angle-closure glaucoma, during an acute attack the pupil-blocking force was markedly high. This technique for measuring pupil-blocking force will provide valuable data for the analysis of the mechanism of primary angle-closure glaucoma.


Feasibility and acute healing of vocal fold microflap incisions in a rabbit model.
The purpose of this study was to investigate the feasibility of performing mucosal elevation of a vocal fold microflap in a rabbit model and to measure the acute healing of rabbit microflap incisions compared to control vocal folds. Prospective animal study. Ten New Zealand white rabbits were used in this study. All rabbits received a 3-mm incision through the epithelium of one vocal fold using a sickle knife and mucosal elevation through this incision using a microlaryngeal fine-angled spatula. The contralateral vocal fold was left intact to serve as an internal control. Student t tests were used to investigate differences in epithelial thickness, immunohistochemical staining of CD45, and inflammatory and profibrotic gene expression between vocal folds undergoing microflap and control. Exposure of the rabbit larynx was achieved, allowing for the identification of a surgical plane and the creation of a microflap and elevation of the vocal fold mucosa. Hematoxylin-and-eosin staining revealed no significant differences in epithelial thickness, immunohistochemistry for CD45 showed no significant differences in CD45-positive cells, and quantitative polymerase chain reaction revealed no significant differences in interleukin-1β, transforming growth factor β-1, or cyclooxygenase-2 gene expression between vocal folds undergoing microflap and control. We demonstrate the feasibility of vocal fold microflap surgery in a rabbit model. With the advantage of greater access to primers and antibodies for molecular biologic studies, the application of the microflap technique in a small-animal model such as rabbit has broad implications for future experimental investigations in laryngology.


Simultaneous removal of triclosan and Cd(Ⅱ) by bio-reduced graphene oxide and its mechanism.
The co-existence of contaminants such as triclosan (TCS) and Cadmium in wastewater is a major public health problem because of their persistence and toxicity. In this study, bio-reduced graphene oxide (B-rGO) synthesized by Lysinibacillus sp. Simultaneously remove TCS and Cd(II), with adsorption capacities of 81.91 and 23.32 mg g-1, for TCS and Cd (Ⅱ), respectively. This was significantly higher than that previously reported for commercially available reduced graphene oxide (C-rGO), which was only 31.94 and 2.01 mg g-1, for TCS and Cd (Ⅱ), respectively. Fourier transform infrared spectroscopy (FTIR) showed that rGO surface-bound extracellular polymeric substances (EPS) played a key role in the observed enhanced contaminant removal, which was verified by scanning electron microscopy (SEM), energy dispersive spectrometry (EDS) and X-ray photoelectron spectroscopy (XPS). In addition, the absorption of both Cd(II) and TCS on B-rGO was confirmed by XPS and high-performance liquid chromatography (HPLC-UV). The adsorption kinetics of both TCS and Cd(II) fitted well to the pseudo-second-order model, while the adsorption isotherms of Cd(II) followed the Langmuir model, and triclosan the Freundlich model. A mechanism of simultaneous removal of TCS was proposed based on π-π interactions and hydrogen bonding, while Cd(II) was removed by a combination of electrostatic and chelation/complexation. Finally, the adsorption of TCS and Cd(II) by B-rGO in real wastewater was shown to be 76.67 and 16.53 mg g-1, respectively, demonstrating that B-rGO has the potential for practical simultaneous removal of TCS and Cd(II) from wastewater.


Technical modifications for transplant in the failing Fontan.
Heart transplant after Fontan completion poses a unique surgical challenge. Twenty patients are presented, stressing the technical hints performed in the five anastomoses to match the graft in the recipient. Data are collected from 20 Fontan patients between 2013 and 2019. Age (13 years), weight (37 kg.), and time interval between Fontan and transplant (7 years) are presented as median. Extracardiac conduit (size 18/20) was implanted in 15 patients, whereas atrio-pulmonary connection was performed in 4 and lateral tunnel in 1. Six patients developed protein-losing enteropathy. Seventeen stents had been previously deployed. The five anastomoses underwent some changes. Left atrium once, aorta 9 times, superior vena cava 7 times, pulmonary branches 15 times, and inferior vena cava 12 times. Follow-up was complete for a median of 42 months (range 6-84). Two patients died. ECMO was needed in six cases for pulmonary hypertension. Four patients had collateral vessels occluded in the cath lab, and stents were placed in superior vena cava (1) and aorta (1) post-transplant. Protein-losing enteropathy was resolved in five patients. Interestingly, one patient was on a systemic assist device before transplant (Levitronix) and right assistance (ECMO) afterwards. Transplant in Fontan patients is actually challenging. Hints in every of the five proposed anastomoses must be anticipated, including stents removal. Extra tissue from the donor (innominate vein, aortic arch, and pericardium) is strongly advisable. ECMO for right ventricular dysfunction was needed in nearly one-third of the cases. Overall results can match other transplant cohorts.


Differential effects of inescapable footshocks and of stimuli previously paired with inescapable footshocks on dopamine turnover in cortical and limbic areas of the rat.
The effect of electric footshocks and of exposure to environmental stimuli paired with electrical shocks upon the dopaminergic activity in various cortical and limbic areas of the rat were evaluated by measuring dihydroxyphenylacetic acid (DOPAC) levels in these areas. In animals exposed to a 20 min electric footshock session DOPAC concentrations were significantly increased in the antero-medial and sulcal frontal cortices, olfactory tubercle, nucleus accumbens and amygdaloid complex (by 66, 37, 28, 55 and 90% respectively). Re-exposure of rats to an environment where they had been shocked 24 h earlier induced an elevation of DOPAC content only in the anteromedial frontal cortex (by 47%). Plasma corticosterone levels were elevated in both situations. No change in serotonin or 5-hydroxyindolacetic acid content of these areas could be detected in either situation. The results show that electric footshocks and environmental stimuli associated to previous shocks both activate central dopaminergic systems, although the patterns of activation are different.


Tyrosinase inhibition: conformational analysis based studies on molecular dynamics calculations of bipiperidine based inhibitors.
Two series of variably N-substituted biperidines were synthesized by condensing various acid chlorides, alkyl halides and anhydrides with 1,4-bipiperidine. The new compounds were tested as tyrosinase inhibitors and a structure-activity relationship (SAR) study was carried out. Potent inhibition was observed in the case of the 4'-methylbenzyl substitution on this atom (IC50 = 1.72 microM) with this compound being a lead for future drug design. Additionally, calculations of the important QSAR molecular descriptors were done on the biperidine analogues after their 2 ps molecular dynamics (MD) simulations using molecular mechanics force field (MMFF) approaches. Using MD simulations potential and total energies were calculated for the energy minimized models of bipiperidine and the most active analogs 2, 3, 4, 6, 8 and 10.


Differential diagnosis of plantar neuromas: a preliminary report.
A discussion has been presented concerning etiology, diagnosis, and treatment of plantar neuromas. The surgeon should, at all times, attempt to ascertain the particular biomechanical fault which predisposed the patient to develop a neuroma, and he should attempt to correct or accommodate the underlying pathology.


Comparison of muscarinic receptor- and beta-adrenoceptor-mediated vasorelaxation between euthyroid and acute hyperthyroid rats.
Hyperthyroidism was induced by subcutaneous injections of L-thyroxine (T4) (0.5 mg/kg/day) for 3 days in order to investigate the effects of acute hyperthyroidism on the vasorelaxing responses to isoprenaline and acetylcholine in isolated rat aortae. In the aortae, there was no significant difference in isoprenaline-induced relaxation between hyperthyroid and control rats, however acetylcholine-induced relaxation was significantly greater in hyperthyroid rats than in control rats. N(G)-nitro-L-arginine (L-NOARG), an inhibitor of nitric oxide (NO) synthase, reduced isoprenaline- and acetylcholine-induced relaxations in both hyperthyroid and control rats and in the presence of L-NOARG no significant difference in the acetylcholine-induced relaxation was seen between the two groups of rats. Indomethacin, a cyclo-oxygenase inhibitor, had no significant influence on both isoprenaline- and acetylcholine-induced relaxations in both control and hyperthyroid rats. 17-Octadecynoic acid (17-ODYA), a cytochrome P-450 mono-oxygenase inhibitor, reduced the both isoprenaline- and acetylcholine-induced relaxation in both hyperthyroid and control rats, and acetylcholine-induced relaxation was still greater in hyperthyroid rats than in control rats. These results indicate that an acute hyperthyroidism significantly enhances muscarinic receptor- but not adrenoceptor-mediated relaxations of the aortae and L-NOARG abolished an enhancement by acute hyperthyroidism of muscarinic receptor-mediated relaxation, suggesting that the effects may be due to an alteration in muscarinic receptor-mediated NO systems of the aortae at early stage of hyperthyroidism.


Endothelins Inhibit Osmotic Swelling of Rat Retinal Glial and Bipolar Cells by Activation of Growth Factor Signaling.
Water accumulation in retinal glial (Müller) and neuronal cells resulting in cellular swelling contributes to the development of retinal edema and neurodegeneration. Here, we show that endothelin-1 (ET-1) dose-dependently inhibits the hypoosmotic swelling of Müller cells in freshly isolated retinal slices of control and diabetic rats, with a maximal inhibition at 100 nM. Osmotic Müller cell swelling was also inhibited by ET-2. The effect of ET-1 was mediated by activation of ETA and ETB receptors resulting in transactivation of metabotropic glutamate receptors, purinergic P2Y1, and adenosine A1 receptors. ET-1 (but not ET-2) also inhibited the osmotic swelling of bipolar cells in retinal slices, but failed to inhibit the swelling of freshly isolated bipolar cells. The inhibitory effect of ET-1 on the bipolar cell swelling in retinal slices was abrogated by inhibitors of the FGF receptor kinase (PD173074) and of TGF-β1 superfamily activin receptor-like kinase receptors (SB431542), respectively. Both Müller and bipolar cells displayed immunoreactivities of ETA and ETB receptor proteins. The data may suggest that neuroprotective effects of ETs in the retina are in part mediated by prevention of the cytotoxic swelling of retinal glial and bipolar cells. ET-1 acts directly on Müller cells, while the inhibitory effect of ET-1 on bipolar cell swelling is indirectly mediated, via stimulation of the release of growth factors like bFGF and TGF-β1 from Müller cells.


Structural connectivity and response to ketamine therapy in major depression: A preliminary study.
Ketamine elicits an acute antidepressant effect in patients with major depressive disorder (MDD). Here, we used diffusion imaging to explore whether regional differences in white matter microstructure prior to treatment may predict clinical response 24h following ketamine infusion in 10 MDD patients. FSL's Tract-Based Spatial Statistics (TBSS) established voxel-level differences in fractional anisotropy (FA) between responders (patients showing >50% improvement in symptoms 24h post-infusion) and non-responders in major white matter pathways. Follow-up regions-of-interest (ROI) analyses examined differences in FA and radial (RD), axial (AD) and mean diffusivity (MD) between responders and non-responders and 15 age- and sex-matched controls, with groups compared pairwise. Whole brain TBSS (p<0.05, corrected) and confirmatory tract-based regions-of-interest analyses showed larger FA values in the cingulum and forceps minor in responders compared to non-responders; complementary decreases in RD occurred in the cingulum (p<0.05). Only non-responders differed from controls showing decreased FA in the forceps minor, increased RD in the cingulum and forceps minor, and increased MD in the forceps minor (p<0.05). Non-responders showed an earlier age of onset and longer current depressive episode than responders. Though these factors did not interact with diffusion metrics, results may be impacted by the limited sample size. Though findings are considered preliminary, significant differences in FA, RD and MD shown in non-responders compared to responders and controls in fronto-limbic and ventral striatal pathways suggest that the structural architecture of specific functional networks mediating emotion may predict ketamine response in MDD.