Effects of paraquat on canine bronchoalveolar lavage fluid.
Bronchoalveolar lavage (BAL) recovers the epithelial lung fluid of the lower respiratory tract. In this study, we have used BAL to detect early pulmonary injury in beagle dogs following an intravenous infusion of 10 mg paraquat dichloride/kg bodyweight. Bronchoalveolar lavage was performed twice in 11 dogs, 60 hr before and 34 hr after an intravenous infusion of paraquat dichloride (n = 8) or saline (n = 3). The dogs were studied in three groups: (1) paraquat only (n = 4); (2) paraquat plus hemoperfusion (n = 4); and (3) hemoperfusion only (n = 3). Because hemoperfusion, a treatment used for paraquat poisoning, could have effects on BAL independent of paraquat, we evaluated the effects on BAL fluid of this procedure performed separately from and together with administration of paraquat. We examined cytology, proteins, enzymes, and glutathione in the BAL fluid and expressed all results per milliliter of aspirated lavage fluid. Hemoperfusion did not alter the BAL fluid. In contrast, in dogs studied 34 hr after administration of paraquat, total cell counts, alveolar macrophage and neutrophil counts, and concentrations of total protein, albumin, ACE, LDH, and ALP were increased. Bronchoalveolar lavage in the dog provides an excellent tool with which to detect early paraquat-induced pulmonary injury. The same technique could be useful for sequential monitoring of other types of pulmonary disease and injury.


Gas Plasma Exposure of Glioblastoma Is Cytotoxic and Immunomodulatory in Patient-Derived GBM Tissue.
Glioblastoma multiforme (GBM) is the most common primary malignant adult brain tumor. Therapeutic options for glioblastoma are maximal surgical resection, chemotherapy, and radiotherapy. Therapy resistance and tumor recurrence demand, however, new strategies. Several experimental studies have suggested gas plasma technology, a partially ionized gas that generates a potent mixture of reactive oxygen species (ROS), as a future complement to the existing treatment arsenal. However, aspects such as immunomodulation, inflammatory consequences, and feasibility studies using GBM tissue have not been addressed so far. In vitro, gas plasma generated ROS that oxidized cells and led to a treatment time-dependent metabolic activity decline and G2 cell cycle arrest. In addition, peripheral blood-derived monocytes were co-cultured with glioblastoma cells, and immunomodulatory surface expression markers and cytokine release were screened. Gas plasma treatment of either cell type, for instance, decreased the expression of the M2-macrophage marker CD163 and the tolerogenic molecule SIGLEC1 (CD169). In patient-derived GBM tissue samples exposed to the plasma jet kINPen ex vivo, apoptosis was significantly increased. Quantitative chemokine/cytokine release screening revealed gas plasma exposure to significantly decrease 5 out of 11 tested chemokines and cytokines, namely IL-6, TGF-β, sTREM-2, b-NGF, and TNF-α involved in GBM apoptosis and immunomodulation. In summary, the immuno-modulatory and proapoptotic action shown in this study might be an important step forward to first clinical observational studies on the future discovery of gas plasma technology's potential in neurosurgery and neuro-oncology especially in putative adjuvant or combinatory GBM treatment settings.


Patterns of Prior and Subsequent Neoplasms in Children and Adolescents With Soft Tissue Sarcomas.
The occurrence of prior, concurrent and subsequent neoplasms (SN) represents a serious problem in children and adolescents with soft tissue sarcomas. Pathogenic germline variants contribute to the diagnosis of multiple neoplasms in sarcoma survivors. The records of 748 children and adolescents, diagnosed with soft tissue sarcomas and registered in trials/registries by the cooperative soft tissue sarcoma (Cooperative Weichteilsarkom Studie) group, were reviewed for the occurrence of SNs. Reference histology review was available for all cases; the presence of oncogenic fusions known at the time of diagnosis was confirmed for fusion-positive (F+) entities. Concurrent or subsequent SNs developed in 13 of 473 survivors of fusion-negative (F-) sarcomas, for an 8-year cumulative SN incidence of 5% in survivors of F- sarcomas. In contrast, only 1 of 278 survivors of F+ sarcoma developed an SN. Twenty of 748 patients with soft tissue sarcomas had a history of prior neoplasms. Six of 14 patients who developed SNs after their index sarcomas met Chompret criteria for Li-Fraumeni syndrome. Nine of 20 patients who had tumors before their index sarcoma diagnosis had neurofibromatosis type 1 or neurofibromatosis type 1 spectrum tumors. Sarcoma phenotype/genotype and the sequence and nature of prior and subsequent neoplasms provide a window into underlying germline genetic susceptibilities in children and adolescents with soft tissue sarcomas.


Cytosine deaminase: structural modifications studies.
Structural modification studies have been shown that a cysteine, a histidine and possibly an arginine residue are involved in the catalytic process. The enzyme gave a single band on polyacrylamide gel electrophoresis, and the amino acid analysis showed it to contain a high proportion of hydrophobic residues, which was in agreement with the chemical modification results.


[Epicutaneous electrical recording for immediate diagnosis of the site of intestinal occlusions and subocclusions].
64 patients have been studied by means of an original abdominal epicutaneous electric recording technique. Cases of intestinal paralysis did not show up waves of particular importance while cases of intestinal occlusion showed waves of three basic types. A close relationship was found between types of waves and occlusion localization. The physiopathological and clinical implications of this diagnostic approach are discussed.


Administration of amyloid-β oligomer to the buccal ganglia may reduce food intake and cholinergic synaptic responses within the feeding neural circuit in Aplysia kurodai.
Anorexia is a behavioral change caused by functional brain disorders in patients with Alzheimer's disease (AD). Amyloid-β (1-42) oligomers (o-Aβ) are possible causative agents of AD that impair signaling via synaptic dysfunction. In this study, we used Aplysia kurodai to study functional disorders of the brain through o-Aβ. Administration of o-Aβ to the buccal ganglia (feeding brain for oral movements) by surgical treatment significantly reduced food intake for at least five days. Furthermore, we explored the effects of o-Aβ on the synaptic function in the feeding neural circuit, focusing on a specific inhibitory synaptic response in jaw-closing motor neurons produced by cholinergic buccal multi-action neurons because we recently found that this cholinergic response decreases with aging, which is consistent with the cholinergic hypothesis for aging. Administration of o-Aβ to the buccal ganglia significantly reduced the synaptic response within minutes, whereas administration of amyloid-β (1-42) monomers did not. These results suggest that o-Aβ may impair the cholinergic synapses, even in Aplysia, which is consistent with the cholinergic hypothesis for AD.


3-(4-Bromo-phenyl-sulfin-yl)-5-cyclo-hexyl-2-methyl-1-benzofuran.
In the title compound, C(21)H(21)BrO(2)S, the cyclohexyl ring adopts a chair conformation. The 4-bromo-phenyl ring makes a dihedral angle of 81.62 (6)° with the mean plane of the benzofuran fragment. In the crystal, mol-ecules are linked by weak C-H⋯O and C-H⋯π inter-actions. The crystal structure also exhibits a slipped π-π inter-action between the furan rings of neighbouring mol-ecules [centroid-centroid distances = 3.540 (3) Å, inter-planar distance = 3.481 (3) Å and slippage = 0.644 (3) Å].


Hemodialysis Access in the Elderly: Outcomes among Patients Older than Seventy.
Arteriovenous fistulae (AVF) are the preferred access for hemodialysis, however, there continues to be limited data guiding the surgical management of primary dialysis access creation in elderly end-stage renal disease patients. This is an 8-year retrospective institutional study evaluating the operative and clinical characteristics of patients 70 years or above who underwent primary, first-ever, AVF or arteriovenous graft (AVG) creation. There was no overall significant difference in AVF and AVG primary (P = 0.127) and secondary (P = 0.870) patency. AVG had higher graft loss secondary to infection (P = 0.0002) and thrombosis (P = 0.0213). Survival was less than 50% at 2 years for AVF and AVG patients. An equal number of AVF and AVG patients who died had functional access at the time of death (P = 1.0000) with more AVG patients using their graft (initiating dialysis) before death (P = 0.0118). Elderly patient patency rates for AVF and AVG are satisfactory and support surgical access creation; however, overall 2-year survival is low. An equal number of AVF and AVG patients died with functioning surgical access, however, more AVG patients initiated dialysis and successfully used their access. Accordingly, special consideration needs to be given with regard to estimated timing until dialysis and predicted patient longevity. Delaying access creation until dialysis is needed and proceeding with AVG creation appears to be justified.


Periodontal status following orthodontic mini-screw insertion: A prospective clinical split-mouth study.
Anchorage control is one of the most important determinants of orthodontic treatments. Mini-screws are used to achieve the desired anchorage. Despite all their advantages, there is a possibility that treatment will not be successful due to conditions related to their interaction with the periodontal tissue. To evaluate the status of the periodontal tissue at the sites adjacent to the orthodontic mini-implants. A total of 34 teeth (17 case and 17 control) in 17 orthodontic patients requiring a mini-screw in the buccal area to proceed with their treatment were included in the study. Oral health instruction was provided to the patients prior to the intervention. In addition, scaling and root planing of the root surface were done using manual instruments and ultrasonic instruments if needed. For tooth anchorage, a mini-screw with Elastic Chain or Coil Spring was used. The following periodontal indices were examined in the mini-screw receiving tooth and the contralateral tooth: plaque index, pocket probing depth, attached gingiva level (AG), and gingival index. Measurements were made before the placement of the mini-screws and 1, 2, and 3 months following that. The results revealed a significant difference only in the amount of AG between the tooth with mini-screw and the control tooth (p = 0.028); for other periodontal indices, there were no significant differences between the two groups. This study showed that periodontal indices in adjacent teeth of the mini-screws do not change significantly compared to other teeth and mini-screws can be used as a suitable anchorage without posing a threat to the periodontal health. Using mini-screws is a safe intervention for orthodontic treatments.


"Bottom-up" meets "top-down" assembly in nanoscale polyoxometalate clusters: self-assembly of [P4W52O178](24-) and disassembly to [P3W39O134](19-).
The pH-controlled assembly/disassembly of a nanoscale {P4W52O178}24- cluster at pH 2 to a {P4W44O152}20- cluster at pH 3-5 via a {P3W39O134}19- cluster species at pH 2-3 to finally give {P2W19O69(OH2)}14- at pH 6 is reported. This process can be traced in the solid state crystallographically and in solution using dynamic light scattering studies.


Using intravoxel incoherent motion MR imaging to study the renal pathophysiological process of contrast-induced acute kidney injury in rats: Comparison with conventional DWI and arterial spin labelling.
To investigate the potential of intravoxel incoherent motion (IVIM) to assess the renal pathophysiological process in contrast-induced acute kidney injury (CIAKI). Twenty-seven rats were induced with CIAKI model, six rats were imaged longitudinally at 24 h prior to and 30 min, 12, 24, 48, 72 and 96 h after administration; three rats were randomly chosen from the rest for serum creatinine and histological studies. D, f, D* and ADC were calculated from IVIM, and renal blood flow (RBF) was obtained from arterial spin labelling (ASL). A progressive reduction in D and ADC was observed in cortex (CO) by 3.07 and 8.62 % at 30 min, and by 25.77 and 28.16 % at 48 h, respectively. A similar change in outer medulla (OM) and inner medulla (IM) was observed at a later time point (12-72 h). D values were strongly correlated with ADC (r = 0.885). As perfusion measurement, a significant decrease was shown for f in 12-48 h and an increase in 72-96 h. A slightly different trend was found for D*, which was decreased by 26.02, 21.78 and 10.19 % in CO, OM and IM, respectively, at 30 min. f and D* were strongly correlated with RBF in the cortex (r = 0.768, r = 0.67), but not in the medulla. IVIM is an effective imaging tool for monitoring progress in renal pathophysiology undergoing CIAKI. • IVIM analysis permits separate quantification of diffusion and perfusion. • IVIM can provide useful biomarkers ifor changes in renal pathophysiology. • IVIM can be useful for monitoring progress in renal pathophysiology undergoing CIAKI.


Infraspinatus muscle hypertrophy and wasting of axillary folds as the important signs in Duchenne muscular dystrophy.
Eighty-four patients with Duchenne muscular dystrophy (DMD) were examined clinically for hypertrophy and wasting in different muscles, parts of the muscles or muscle groups. Some muscles were examined under mild contraction to bring out any subclinical pseudohypertrophy. Findings revealed infraspinatus muscle hypertrophy to be significantly frequent (88%) and closely second to well known calf hypertrophy (94%). Infraspinatus hypertrophy was noted in 5 such DMD patients in whom calf hypertrophy was unremarkable. The wasting was consistently observed in the muscles forming anterior and posterior axillary folds. In conclusion, infraspinatus muscle hypertrophy and wasting of axillary folds are the important supportive clinical evidences during the examination of DMD patients.


Recurrent osteoid osteoma of the index proximal phalanx: A case report with a new resection technique.
Osteoid osteoma is a relatively common benign bone-forming lesion that often occurs in young adults with a predilection for the lower limbs and spine. It is characterized by the formation of osteoid tissue, a precursor to bone, and can cause pain and restricted mobility in affected areas. Osteoid osteoma of the hand and fingers is rare, thus representing a diagnostic challenge. A written informed consent was provided from the patient's father for the publication and sharing all images and clinical data concerning the patient. We are reporting a case of an 11-year-old girl who presented with persistent pain and swelling in her left index finger, as well as limited mobility, over the course of 1 year. Nonsteroidal anti-inflammatory drugs eased the pain, but it resurfaced once the medication was discontinued. A series of investigations, showed a lytic lesion at the second proximal phalangeal neck, with features indicative of osteoid osteoma. Excision with bone grafting was performed as definitive therapy with pathological confirmation of the osteoid osteoma diagnosis. Nevertheless, the patient returned 2 years later with a recurrence of her previous symptoms, and further tests suggested a recurrence of osteoid osteoma. To address the recurrence, a modified open thermoablation technique was used. Thermoablation is a minimally invasive procedure that uses heat to destroy the tumor cells, and it has been shown to be effective in treating osteoid osteoma. The modified open approach involves making a small incision to access the tumor and delivering heat directly through a previously CT-guided inserted Kirschner wire to the affected area. The patient reported no pain at 1 month and 1 year after the surgery, with no radiological signs of recurrence, indicating complete excision of the lesion. Overall, this case highlights the challenges of diagnosing and treating osteoid osteoma in the hands and fingers. Further research is needed to better understand the underlying causes, potential risk factors, and optimal treatment for osteoid osteoma recurrence.


Detection of circulating tumor lysate-reactive CD4+ T cells in melanoma patients.
We wanted to study whether an allogeneic melanoma lysate would be a feasible stimulatory antigen source for detection of a peripheral CD4+ T-cell immune response in patients with medically untreated malignant melanoma. The lysate was produced from a melanoma cell line (FM3.29) which expresses high amounts of melanoma antigens. Fresh peripheral blood was incubated with and without lysate for 6 h in the presence of anti-CD28/anti-CD49d MoAb (for costimulation). After flow cytometric estimation of the frequency of CD69+/IFN-gamma+ cells in the CD4+ population, the response to lysate was calculated as the difference between the number of activated IFN-gamma-producing CD4+ cells in the lysate-stimulated and the nonstimulated sample. An immune response to lysate was observed in blood samples from 11 of 15 patients (73%) with metastatic melanoma. A weak response was found in 1 of 4 patients radically operated for localized disease, whereas no responders were seen among 7 healthy donors. The fraction of circulating lysate-activated T cells ranged from 0.0037% to 0.080% of the CD4+ population. A negative result of the assay was found occasionally, especially in donors with high background levels of spontaneous IFN-gamma production, indicating an inhibitory effect of the lysate. This method for detection of a peripheral T-cell immune response in melanoma patients has several advantages for clinical use. The tumor lysate preparations may contain large numbers of stimulating antigens (known, as well as unknown) and are easily prepared and handled. Potentially, the assay might be useful as a diagnostic tool, a marker of residual or recurrent disease, a prognostic factor, or a predictor or monitor of the effect of antineoplastic therapy including immune-modulating therapy.


Sambucus Williamsii Hance Promotes MC3T3-E1 Cells Proliferation and Differentiation via BMP-2/Smad/p38/JNK/Runx2 Signaling Pathway.
The 50% ethanol elution fractions of root-bark of Sambucus Williamsii Hance (rbSWH) evaluated the effect of proliferation and differentiation on preosteoblast MC3T3-E1 cell, and the mechanism of actions. We found that rbSWH(30, 60, and 90 µg/mL) can enhance cell proliferation by MTT assay and promote alkaline phosphatase (ALP) and bone Gla protein (BGP) activities, type I collagen (Col-I) synthesis, and mineralization nodule formation in primary cultured osteoblasts. The results showed that rbSWH can increase mRNA levels of BMP-2 and Runx2 using real-time reverse transcription-quantitative polymerase chain reaction, whereas the BMP-2 antagonist Noggin attenuated the increase of ALP activity induced by rbSWH, indicating that BMP-2 expression was required for the action of rbSWH in osteoblastic. We also found that rbSWH can enhance the expressions of BMP-2, BMPRIB, BMPRII, phosphorylation of Smad, JNK and p38, and Runx2 proteins by western blotting. In addition, pretreatment of cells with p38 inhibitor (SB203580) or JNK inhibitor (SP600125) can antagonize the elevation of BMP-2 expression, ALP activity, and cell viability induced by rbSWH. Taken together, our results provided an evidence that rbSWH can promote MC3T3-E1 cell proliferation and differentiation via BMP-2/Smad/p38/JNK/Runx2 signaling pathway.


Retinoic acid receptor fusion proteins: friend or foe.
Acute promyelocytic leukemia (APL) is characterized by a block in differentiation of hematopoietic cells at the promyelocytic stage of development. This disease is uniquely sensitive to treatment with pharmacological doses of all-trans retinoic acid (ATRA), and the combination of ATRA with chemotherapy has improved the durable disease-free survival in these patients to up to 80%. APL is characterized by chromosomal translocations that lead to the fusion of the retinoic acid receptor-alpha (RARalpha) to various partner genes. RARalpha functions as a ligand-inducible transcription factor and the aberrant RARalpha fusion proteins contribute to leukemic transformation by dominant inhibition of the expression of target genes that are important for cellular differentiation. This may at least in part be explained by an abnormally strong interaction with corepressor proteins leading to deacetylation of DNA and silencing of target genes. Most RARalpha fusion proteins can still be induced to transactivate genes, but only at very high doses of ligand, explaining why pharmacological doses of ATRA are necessary to obtain a therapeutic effect in these patients.


Stomatal responses to humidity of coastal and interior populations of a Californian shrub.
Plants of two populations of Diplacus aurantiacus, a subshrub of the Californian chaparral, were compared for their stomatal response to water vapor concentration gradients. Plants of a coastal and an interior population were compared when grown under both low and high humidities. When grown at high humidity the coastal plants exhibited higher conductances and higher transpiration/photosynthesis ratios at all leaf-to-air water vapor concentration gradients than did the interior plants. Although all of the plants examined showed a pronounced stomatal response to humidity the response did not result in the degree of regulation of water-use efficiency reported for other Californian coastal species.


Practical aspects of using methacrylate-ester-based monolithic columns in capillary electrochromatography.
Methacrylate-ester-based monoliths containing quaternary ammonium groups were prepared in situ in capillary columns and in simultaneous experiments in vials, employing thermal initiation. The chromatographic properties of the monoliths were determined with capillary electrochromatography (CEC), and their morphology was studied with mercury-intrusion porosimetry on the bulk materials. Materials with different, well repeatable pore-size distributions could be prepared. A satisfactory column-to-column and run-to-run repeatability was obtained for the electro-osmotic mobility, the retention characteristics (k-values) and the efficiency on the columns prepared and tested in the CEC mode. A relatively high electro-osmotic flow was observed in the direction of the positive electrode. The electro-osmotic mobility was found to be influenced only marginally by mobile-phase parameters such as the pH, ionic strength, and acetonitrile content. The retention behavior of the monolithic columns was similar to that of columns packed with C18-modified silica particles. Columns could be prepared with optimum plate heights ranging from 6 microm for unretained compounds to 20 microm for well retained (k=2.5) polyaromatic hydrocarbons. However, for specific analytes a - still unexplained - lower chromatographic column efficiency was observed.


Rapid on-site evaluation of endoscopic ultrasound core biopsy specimens has excellent specificity and positive predictive value for gastrointestinal lesions.
Endoscopic ultrasound (EUS) with fine needle aspiration (FNA) is a safe and effective way to sample lesions in the gastrointestinal tract. Rapid on-site specimen evaluation (ROSE) improves the accuracy of EUS-FNA. While data suggests that EUS with fine-needle biopsy (EUS-FNB) is effective, it remains unclear if ROSE is predictive of a final diagnosis when obtaining core specimens. The aim of this study was to investigate the utility of ROSE in achieving a final diagnosis for EUS-FNB core specimens. We evaluated 60 consecutive patients referred for EUS guided sampling of lesions within or adjacent to the gastrointestinal tract. All patients underwent EUS-FNB to evaluate the additive value of ROSE to the diagnostic accuracy of specimens obtained using a core biopsy needle. EUS-FNA was also performed in a majority of cases. EUS-FNB was feasible in all 60 cases; on-site specimen adequacy and final diagnostic accuracy was 58 % [95 % confidence intervals (CI) 45.1-71.2] and 83 % (95 % CI 71.9-91.5), respectively. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value of ROSE for core biopsies were 65, 100, 100, and 39 %, respectively. On-site adequacy and diagnostic accuracy for EUS-FNA was 38 % (95 % CI 22.2-53.5) and 63 % (95 % CI 50.1-75.8), respectively. There were no significant complications. EUS-FNB is safe, feasible, and effective. ROSE of the core biopsy provides excellent PPV; however, an inadequate ROSE appears to be of limited value. Further prospective studies are needed to assess the optimal handling and onsite processing of core specimens to determine whether ROSE is beneficial for EUS-FNB.


Effect of Pod e-Cigarettes vs Cigarettes on Carcinogen Exposure Among African American and Latinx Smokers: A Randomized Clinical Trial.
Fourth-generation nicotine salt pod system (NSPS) electronic cigarettes (e-cigarettes) are the leading class of e-cigarettes. They contain high nicotine concentrations, which may facilitate switching among smokers, but could also lead to increased exposure to nicotine and biomarkers of potential harm. African American and Latinx smokers experience significant tobacco-related health disparities. The potential of NSPS e-cigarettes to reduce smoking-related harm among these groups is unknown. To compare the harm reduction potential of NSPS e-cigarette vs combustible cigarettes. This unblinded randomized clinical trial compared 6 weeks of e-cigarette use vs cigarettes as usual from to 2018 to 2019 among smokers in the San Diego, California, and Kansas City, Missouri, areas. Participants included African American and Latinx adult combustible cigarette smokers who smoked at least 5 cigarettes/d on at least 25 of the past 30 days for at least 6 months and were interested in switching to e-cigarettes. Data were analyzed from September 18, 2019, to September 4, 2020. 6 weeks of e-cigarette use in a choice of pod flavors (5% nicotine) along with brief education, training, and action planning to completely switch to e-cigarettes from combustible cigarettes. The control group smoked combustible cigarettes as usual. The primary outcome was reduction in urinary 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) concentration at week 6. Secondary outcomes were change in urinary cotinine, expired carbon monoxide (CO), respiratory symptoms, lung function, blood pressure, past 7-day consumption of combustible cigarettes, and switching rates (e-cigarette group only) at weeks 2 and 6. This study included 186 participants, including 92 African American participants and 94 Latinx participants. The mean (SD) age was 43.3 (12.5) years, and 75 (40.3%) were women. Participants smoked a mean (SD) of 12.1 (7.2) cigarettes/d on 6.8 (0.6) d/wk at baseline. A total of 125 participants were randomized to the e-cigarette group and 61 were randomized to the control group. At baseline, median (interquartile range) NNAL was 124 (45-197) pg/mL in the e-cigarette group and 88 (58-197) pg/mL in the control group. At week 6, the e-cigarette group had significantly greater reductions in NNAL (relative risk [RR], 0.36 [95% CI, 0.23-0.54]; P < .001), CO (RR, 0.53 [95% CI, 0.42-0.68]; P < .001), respiratory symptoms (RR, 0.63 [95% CI, 0.47-0.85]; P = .002), and number of cigarettes smoked in the past 7 days among those still smoking (RR, 0.30 [95% CI, 0.20-0.43]; P < .001) than the control group and maintained their cotinine levels (RR, 0.80 [95% CI, 0.58-1.10]; P = .17). Lung function and diastolic and systolic blood pressure remained unchanged and did not differ between groups. For participants randomized to receive e-cigarettes, 32 participants (28.1%) were exclusively using e-cigarettes at week 6, while 66 participants (57.9%) were dual using and 16 participants (14%) resumed exclusively using cigarettes. These findings suggest that e-cigarettes may be an inclusive harm reduction strategy for African American and Latinx smokers. ClinicalTrials.gov Identifier: NCT03511001.


Unraveling proteome changes and potential regulatory proteins of bovine follicular Granulosa cells by mass spectrometry and multi-omics analysis.
In previous study, we performed next-gene sequencing to investigate the differentially expressed transcripts of bovine follicular granulosa cells (GCs) at dominant follicle (DF) and subordinate follicle (SF) stages during first follicular wave. Present study is designed to further identify the key regulatory proteins and signaling pathways associated with follicular development using label-free liquid chromatography-tandem mass spectrometry (LC-MS/MS) and multi-omics data analysis approach. DF and SF from three cattle were collected by daily ultrasonography. The GCs were isolated from each follicle, total proteins were digested by trypsin, and then proteomic analyzed via LC-MS/MS, respectively. Proteins identified were retrieved from Uniprot-COW fasta database, and differentially expressed proteins were used to functional enrichment and KEGG pathway analysis. Proteome data and transcriptome data obtained from previous studies were integrated. Total 3409 proteins were identified from 30,321 peptides (FDR ≤0.01) obtained from LC-MS/MS analysis and 259 of them were found to be differentially expressed at different stage of follicular development (fold Change > 2, P < 0.05). KEGG pathway analysis of proteome data revealed important signaling pathways associated with follicular development, multi-omics data analysis results showed 13 proteins were identified as being differentially expressed in DF versus SF. This study represents the first investigation of transcriptome and proteome of bovine follicles and offers essential information for future investigation of DF and SF in cattle. It also will enrich the theory of animal follicular development.


Improving measurement of harms from others' drinking: A key informant study on type and severity of harm.
Some types of harms experienced because of others' drinking (AHTO) may produce greater negative effects than other harms. However, AHTO survey items were developed to measure type, not severity, of harm. We aimed to compare the perceived severity of a comprehensive list of AHTO items to assess consistency in subjective ratings of severity, facilitate a more nuanced analysis and identify strategies to improve measurement of AHTO in epidemiological surveys. Thirty-six leaders of national alcohol surveys (conducted between 1997 and 2016) from 23 countries rated the typical severity of negative effects on the victim of each of 48 types of AHTO using a scale from zero (no negative effect) to 10 (very severe negative effect). The survey leaders were also asked to provide open-ended feedback about each harm and the severity-rating task generally. Of 48 harm items, five were classified as extreme severity (mean rating ≥8), 17 as high (≥6 <8), 25 as moderate (≥4 <6), and one as low (≤4). We used two-way random effects models to estimate absolute agreement intraclass correlation coefficients (AA-ICC) and consistency of agreement intraclass correlation coefficients (CA-ICC). Results showed that there was fair to excellent absolute agreement and consistency of agreement among "experts'' ratings of the severity of harms from others' drinking (single measures CA-ICC = 0.414, single measures AA-ICC = 0.325; average CA-ICC = 0.940, average AA-ICC = 0.914). Harms to children, and harms causing physical, financial, practical, or severe emotional impacts were rated most severe. When designing new AHTO surveys and conducting analyses of existing data, researchers should pay close attention to harms with high perceived severity to identify effective ways to prevent severe AHTO and reduce the negative health and social impacts of AHTO. In-depth analyses of specific sub-sets of harms and qualitative interviews with victims of severe AHTO may prove useful.


Imprints of pandemic lockdown on subsurface water quality in the coastal industrial city of Tuticorin, South India: A revival perspective.
Globally, the incidences of environmental improvements owing to seizing the anthropogenic activities during the lockdown have been reported through news articles and photographs, yet a formal scholarly study has been lacking to substantiate the imprints of lockdown. We hereby present the imprints of lockdown on water quality (both chemical and biological) parameters during the nationwide lockdown (COVID-19 epidemic) in India between 25th March to 30th May 2020. The present study describes the changes in chemical and biological water quality parameters based on twenty-two groundwater samples from the coastal industrial city of Tuticorin in Southern India, taken before (10 and 11th February 2020) and during the lockdown (19 and 20th April 2020) periods. The physico-chemical parameters compared are pH, total dissolved solids (TDS) and electrical conductivity (EC), nitrate (NO3), fluoride (F), chromium (Cr), iron (Fe), copper (Cu), zinc (Zn), cadmium (Cd), lead (Pb), arsenic (As), and selenium (Se), and the bacterial parameters are total coliforms, fecal coliforms, E. coli, and fecal streptococci. Among the metals, the significant reductions in Se (42%), As (51%), Fe (60%) and Pb (50%) were noticed probably owing to no or very less wastewater discharges from metal-based industries, seafood-based industries and thermal power plants during the lockdown. Reduction in NO3 (56%), total coliform (52%) and fecal coliforms (48%) indicated less organic sewage from the fishing industries. Contents of Cr, Cu, Zn and Cd, however, remained similar and fluoride did not show any change, probably as they were sourced from rock-water interactions. Similarly, we did not observe alterations in E. coli and fecal streptococci due to no significant change in domestic sewage production during the lockdown. The multivariate analyses aptly illustrated this and the principal component analyses helped to identify the sources that controlled water qualities of the lockdown compared to the pre-lockdown period. Our observation implies that groundwater is definitely under active interaction with surface waters and thus a quick revival could be observed following the seizing of anthropogenic activities.


Role of positron emission tomography scanning in the management of lung nodules detected at baseline computed tomography screening.
Indeterminate noncalcified lung nodules are a frequent finding when low-dose computed tomography (LD-CT) is used for lung cancer screening. The best clinical management for such nodules remains uncertain. We present results using positron tomography scanning (CT-PET) to evaluate LD-CT-detected lung nodules during the first year of the Continuing Observation of Smoking Subjects (COSMOS) early detection trial for lung cancer. A total of 5200 asymptomatic current or former smokers (> or = 20 pack-years) older than 50 years of age were enrolled in a single-institution screening trial using annual LD-CT. Growing nodules and those with a maximum diameter exceeding 8 mm were studied with CT-PET. Transthoracic needle biopsy was not a routine part of the protocol. During the first year of study, 157 subjects underwent CT-PET, 66 of whom underwent surgical biopsy. Of the 58 lung cancers found on surgical biopsy, 51 were positive (standard uptake value > 2.0) and seven were negative for malignancy by CT-PET. Sensitivity was 88% overall, but 100% in the subgroup with solid nodules of 10 mm or more. Among the 8 patients with benign disease at surgical biopsy, CT-PET was positive in 6 and negative in 2. CT-PET is a highly promising modality for identifying potentially malignant lesions in screening-detected lung nodules and appears particularly useful as an alternative, in the screening setting, to invasive procedures for the further investigation of uncertain nodules. Our findings also indicate that the standard uptake value threshold for positivity should be lowered for small nodules (< 10 mm). Longer follow-up and larger prospective studies are necessary to confirm these preliminary findings.


Effects of genetic resistance against Herpes simplex virus in vaginally infected mice.
In order to take the conditions of natural Herpes simplex virus (HSV) infection into consideration, the genetic resistance of C57-Bl mice, which was established by intraperitoneal HSV-1 infection [Lopez, 1975], was investigated in vaginally infected mice. The course of infection in the mucous membranes did not differ in sensitive (NMRI) and resistant (C57-Bl) mice: both number of takes and virus elimination from the vagina were equal, and no difference in viral titer produced in the vagina was detected. Viral titer in the productively infected lumbosacral ganglia, however, was less in the resistant mice. An experiment with foot-pad-infected mice confirmed that the number of productively infected ganglia was reduced in resistant mice, and contralateral ganglia were infected only in the sensitive mouse strain. In spite of this, the number of latently infected animals did not vary significantly in the mouse strains. Higher activity of defense mechanisms in resistant mice, apparently localized in the ganglia, resulted in reduced lethality. As to the mechanisms of the resistance, neither antibody nor interferon response were enhanced in C57-Bl mice, but resistance was abolished by depletion of several cellular functions, i.e., lymphocytes by cyclophosphamide or X-rays, macrophages by silica or macrophage-antiserum, and M-cells by 89Sr.


Hypolipidemic effects in dogs of ML-236B, a competitive inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase.
ML-236B, a competitive inhibitor of 3-hydroxy-3-methylglutaryl-CoA reductase, significantly reduced both serum cholesterol and phospholipid levels in dogs, when used at a dosage higher than 10 mg/kg per day. Triglyceride levels were not consistently changed, but beta- and pre-beta-lipoproteins were preferentially reduced. Serum cholesterol levels were reduced by 44--45% at the higher dosage of 100--400 mg/kg per day (for 5 weeks) but ML-236B caused no significant changes in the cholesterol content of the liver and aorta and in the activities of serum GOT, GPT, CPK and lecithin : cholesterol acyltransferase. Fecal excretion of neutral sterols was unaffected but that of bile acids was markedly elevated by the drug. Under these conditions, hepatic cholesterol 7alpha-hydroxylase, the rate-limiting enzyme in bile acid biosynthesis, showed no detectable changes.


Meta-analysis of risk factors for Parkinson's disease dementia.
Parkinson's disease (PD) is a common heterogeneous neurodegenerative disorder in elder population. Parkinson's disease dementia (PDD) is one of the most common non-motor manifestations in PD patients. No comprehensive review has been conducted to assess risk factors for PDD. A systemic search for studies on PDD risk factors was performed. Cohort and case-control studies that clearly defined PDD and presented relevant data were included. The data were analyzed to generate a pooled effect size and 95 % confidence interval (CI). Publication bias was assessed using the Egger's test and the Begg's test. A systematic search was conducted and yielded 5195 articles. After screening, 25 studies were included in the current analysis. Development of PDD was positively associated with age (odds ratio [OR] 1.07, 95 % CI 1.03-1.13), male (OR 1.33, 95 % CI 1.08-1.64), higher Unified Parkinson's Disease Rating Scale (UPDRS) part III scores (relative risk [RR] 1.04, 95 % CI 1.01-1.07), hallucination (OR 2.47, 95 % CI 1.36-4.47), REM sleep behavior disorder (RBD) (OR 8.38, 95 % CI 3.87-18.08), smoking (ever vs. never) (RR 1.93, 95 % CI 1.15-3.26) and hypertension (OR 1.57, 95 % CI 1.11-2.22). An inverse association was found between education (RR 0.94, 95 % CI 0.91-0.98) and PDD. Other reported factors, including age of onset, disease duration of PD, Hoehn and Yahr stage and diabetes mellitus were not significantly associated with PDD. Advanced age, male, higher UPDRS III scores, hallucination, RBD, smoking and hypertension increase the risk of PDD, whereas higher education is a protective factor for PDD.


The use of alternative therapies by children with cancer.
To identify the nature and prevalence of therapies used by children with cancer as alternatives to orthodox medical treatment. A descriptive study in which the parents of children with cancer completed a questionnaire describing the use of alternative therapies by their children. Children diagnosed with cancer, excluding brain tumours, at the Women's and Children's Hospital, South Australia, from July 1988 to July 1992. Approximately 46% of the children with cancer had used at least one alternative therapy. In general, parents reported that the therapies were used as an addition to orthodox medical treatment rather than as an alternative. Less than 50% of the parents had discussed the use of the alternative therapies with the children's doctors. A substantial proportion of children with cancer use alternative therapies and the use of these therapies is often not discussed with the children's medical practitioners. There is a continuing need to consider how to better provide, within the context of orthodox medical treatment, those elements of their children's care which parents currently seek from alternative therapies.


A 3D-2D Hybrid U-Net Convolutional Neural Network Approach to Prostate Organ Segmentation of Multiparametric MRI.
Prostate cancer is the most commonly diagnosed cancer in men in the United States with more than 200,000 new cases in 2018. Multiparametric MRI (mpMRI) is increasingly used for prostate cancer evaluation. Prostate organ segmentation is an essential step of surgical planning for prostate fusion biopsies. Deep learning convolutional neural networks (CNNs) are the predominant method of machine learning for medical image recognition. In this study, we describe a deep learning approach, a subset of artificial intelligence, for automatic localization and segmentation of prostates from mpMRI. This retrospective study included patients who underwent prostate MRI and ultrasound-MRI fusion transrectal biopsy between September 2014 and December 2016. Axial T2-weighted images were manually segmented by two abdominal radiologists, which served as ground truth. These manually segmented images were used for training on a customized hybrid 3D-2D U-Net CNN architecture in a fivefold cross-validation paradigm for neural network training and validation. The Dice score, a measure of overlap between manually segmented and automatically derived segmentations, and Pearson linear correlation coefficient of prostate volume were used for statistical evaluation. The CNN was trained on 299 MRI examinations (total number of MR images = 7774) of 287 patients. The customized hybrid 3D-2D U-Net had a mean Dice score of 0.898 (range, 0.890-0.908) and a Pearson correlation coefficient for prostate volume of 0.974. A deep learning CNN can automatically segment the prostate organ from clinical MR images. Further studies should examine developing pattern recognition for lesion localization and quantification.


[Analgesic effect of the selective noradrenaline reuptake inhibitor reboxetine].
Reboxetine is a selective noradrenaline reuptake inhibitor (NARI). As noradrenaline plays a relevant role in antinociceptive mechanisms, the hypothesis was investigated in this study whether reboxetine has analgetic efficacy. Twenty-four healthy volunteers were investigated in a crossover design. Reboxetine (2 x 2 mg) or placebo were given for 5 days with the "crossover" following a one-day washout in between. The primary end-points were the N1 and P2 amplitudes of laser evoked somatosensory potentials (laser SEP) of vertex EEG. In addition, visual analogue scales (VAS, 100 mm) were used. Reboxetine showed a higher analgesic potency than placebo, with a statistically significant superiority in capsaicinirritated skin--in the objective tests (laser SEP, P < 0.0001 for N1 and P = 0.0002 for P2) as well as in the subjective pain measurements (VAS, P < 0.0003). Besides pain, the subjective feelings of burning and itching were lower with reboxetine than with the placebo. The hypothesis that a selective noradrenergic substance could be of analgesic efficacy was proven the first time experimentally. Reboxetine reduced the N1 and the P2 components of the laser SEP, which speaks for central and peripheral mechanisms of action.


An empirical exploration of the relations between the health components of the International Classification of Functioning, Disability and Health (ICF).
The aim of this study was to investigate the relations between the ICF components from a subjective perspective. Data on health condition and perceived functioning were collected among 2941 individuals with at least one chronic disease or disorder. Path analysis was used with perceived level of participation as the final denominator. Three models were tested: one with the number of chronic diseases and disorders as an indicator of health condition, one with perceived health as indicator of health condition, and one with perceived health as part of the personal factors. Although all models showed a good fit, the model with the best fit was that with perceived health as an indicator of health condition. From a patient's perspective, components of the ICF scheme appear to be associated with each other, with perceived health being the best indicator of the health condition.


In vitro activation of ovarian cortex and autologous transplantation: A novel approach to primary ovarian insufficiency and diminished ovarian reserve.
Primary ovarian insufficiency (POI) and diminished ovarian reserve are two conditions that affect women's fertility. Oocyte donation remains an option for these patients; however, the development of certain novel technologies, such as in vitro activation of ovarian cortex (IVA), enables the possibility of activating the pool of resting primordial follicles, increasing the chance of pregnancy. Here, we review the main pathways (PI3K and Hippo signaling) that govern the activation of primordial follicles and its application through the development of culture systems that support ovarian cortex for autologous transplantation. We also review the available data from case reports regarding outcomes of pregnancy and live birth rates with IVA. A PubMed search was conducted using the PubMed-NCBI database to identify literature pertinent to the pathways involved in the activation of primordial follicles and the outcomes of IVA techniques from 2013 to the present. Women with POI have around a 5% chance of spontaneous pregnancy. Recently, novel techniques involving the activation of primordial follicles through molecular pathways have been developed, thus increasing the odds of these patients. More recently, the introduction of a drug-free IVA technique has shown to increase the number of antral follicles with successful oocyte maturation after gonadotropin treatment, reaching pregnancy rates over 30%, either through spontaneous conception or by the implementation of assisted reproductive technology. The evidence of this review is based on a few small series, so data should be interpreted with caution, and only randomized controlled trials could estimate the real magnitude and success of the procedure. IVA technique remains an experimental strategy, with limited available data and the requirement of invasive procedures. Moreover, possible carcinogenic effects not yet determined after transplantation require special caution. In view of the results achieved, IVA could provide a promising option for the preservation of fertility in some cancer patients and prepuberal girls where the only alternative is tissue cryopreservation. The authors received no specific funding for this work and declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.


NAB-1 instructs synapse assembly by linking adhesion molecules and F-actin to active zone proteins.
During synaptogenesis, macromolecular protein complexes assemble at the pre- and postsynaptic membrane. Extensive literature identifies many transmembrane molecules sufficient to induce synapse formation and several intracellular scaffolding molecules responsible for assembling active zones and recruiting synaptic vesicles. However, little is known about the molecular mechanisms coupling membrane receptors to active zone molecules during development. Using Caenorhabditis elegans, we identify an F-actin network present at nascent presynaptic terminals and required for presynaptic assembly. We unravel a sequence of events whereby specificity-determining adhesion molecules define the location of developing synapses and locally assemble F-actin. Next, the adaptor protein NAB-1 (neurabin) binds to F-actin and recruits active zone proteins SYD-1 and SYD-2 (liprin-α) by forming a tripartite complex. NAB-1 localizes transiently to synapses during development and is required for presynaptic assembly. Altogether, we identify a role for the actin cytoskeleton during presynaptic development and characterize a molecular pathway whereby NAB-1 links synaptic partner recognition to active zone assembly.


The role of recombination in the evolvement of the fragile X mutation.
The frequency of recombination in the regions adjacent to the fragile X locus was studied in two groups of carriers: daughters of transmitting males and transmitters of maternally inherited fragile X chromosomes. Approximately one-half of the offspring of the former and one quarter of the offspring of the latter are recombinant. Recombinants and parentals are equally distributed among affected and normal offspring in the two groups. These results indicate that crossing-over at or around the fragile X locus occurs in every meiosis in daughters of transmitting males, although the recombinant chromatids do not necessarily carry the fragile X mutation. Hence, crossing-over is unequivocally associated with, but is not the direct cause of, the transition from the primary genetic lesion to the final mutation.


Differences in physician-reported practice in palliative sedation therapy.
Although palliative sedation therapy is often required in terminally ill cancer patients, little is known about actual practice. The aims of this study were to clarify the physician-reported sedation practices and the factors influencing the sedation rates. A questionnaire was sent to 105 representative physicians of all certified palliative care units in Japan. A total of 81 responses were analyzed (effective response rate, 80%). The prevalence of continuous-deep sedation for physical symptoms was <10% in 33 institutions (41%), 10-50% in 43 institutions (53%), and >50% in 5 institutions (6.2%). The prevalence of continuous-deep sedation for psychoexistential suffering was 0% in 52 institutions (64%), 0.5-5% in 26 institutions (32%) and more than 10% in 3 institutions (3.6%). Continuous-deep sedation was more frequently performed by physicians who did not believe clear consciousness was necessary for a good death, who did not believe that sedation often shortened patient life, who worked with nurses specializing in cancer/palliative care, who judged the symptoms as refractory without actual trials of treatments, who performed continuous sedation first rather than intermittent sedation, and who used phenobarbitones frequently. Physician-reported practice in palliative sedation therapy varied widely among institutions. The differences were mainly associated with the physicians' philosophy about a good death, physicians' belief about the effects of sedation on patient survival, and physicians' medical practice. Discussion should be focused on these divergent areas, and clear clinical guidelines are urgently needed to provide valid end-of-life care.


Serum beta-lipoprotein, serum cholesterol and Quetelet's index as predictors for survival of breast cancer patients.
We studied the survival of breast cancer patients in relation to serum cholesterol level, serum beta-lipoprotein level (BLP) and being overweight among women having breast cancer diagnosed during a follow-up period of 20 years. A cohort of 46,570 women attended a general health screening including examination of serum lipid levels, height and weight during 1963-1965. Of these, 1170 women developing breast cancer; 196 were below the age of 50 and 974 were above 50 years of age. 66 of the younger women, and 341 of the older women were reported to have died of breast cancer. A correlation was found between high serum BLP and decreased survival of breast cancer patients < 50 years of age. For women > or = 60 years of age, BLP was positively correlated to breast cancer survival. No correlation was found between serum cholesterol level and breast cancer survival in any age group. Increasing obesity was statistically significantly correlated to decreased survival with breast cancer. The latter findings were in accordance with other studies which have shown being overweight as a risk factor for breast cancer. As for the relationships between ischaemic heart disease and serum lipid levels, in studies of cancer risks in relation to serum cholesterol level, the different fractions of cholesterol seem to be of importance.


Assessment of pneumoconiosis in surface coal miners after implementation of a national radiographic surveillance program, United States, 2014-2019.
Pneumoconiosis can occur in surface coal miners. The Coal Workers' Health Surveillance Program (CWHSP) has only included surface coal miners as part of its regular disease surveillance since 2014. This analysis identifies the prevalence of pneumoconiosis among working surface coal miners participating in the CWHSP since their initial inclusion, through 2019. Working surface coal miners who had chest radiographs through the CWHSP from January 1, 2014 through December 31, 2019 were included in this analysis. Demographic information, mining tenure and occupation, and radiographic classifications according to the International Labour Office system were included from each miner's most recent encounter with the CWHSP. Prevalence ratios were calculated comparing the prevalence of the disease by region and occupation by log-binomial regression. Pneumoconiosis was present in 109 (1.6%) surface coal miners, including 12 miners with progressive massive fibrosis, the most severe form of the disease. After taking surface mining tenure into account, surface miners in Central Appalachia (prevalence ratio [PR], 3.2; 95% confidence interval [CI], 2.2-4.7) and surface miners who worked as a driller or blaster (PR, 2.1; 95% CI, 1.3-3.5) were at increased risk of pneumoconiosis. The occurrence of pneumoconiosis in surface coal miners supports including them within a systematic respiratory health surveillance program. The current surveillance findings are consistent with past findings of pneumoconiosis, particularly silicosis, in surface mining occupations such as drilling and blasting.


[The graphic representation of the temperature field of the skin surface in the orbital area].
The paper describes results of researches on the possibility of graphic presentation of a temperature field of skin surface of the orbital area. A method of liquid crystal thermography and electron thermometry was used for examination of 30 patients with practically healthy eyes. It was shown that graphic presentation of data of liquid crystal thermography and electron thermometry remarkably facilitates documentation and analysis of the results obtained.


Two Asian families with gastric adenocarcinoma and proximal polyposis of the stomach successfully treated via laparoscopic total gastrectomy.
Family 1: a 39-year-old woman and her sister were admitted to our hospital for fundic gland polyps (FGPs). Their mother died of gastric cancer with FGPs. We performed repeated biopsies at close intervals, suspecting gastric adenocarcinoma and proximal polyposis of the stomach (GAPPS). After a 1-year follow-up, the sisters were diagnosed with gastric cancer with FGP. We performed laparoscopic total gastrectomies with D1+lymph node dissection. Promoter 1B (exon 1B) of the APC gene (chr5: 112,043,224 T>C) contained a point mutation. The sisters were subsequently diagnosed with GAPPS as per the mutational analysis. Family 2 (unrelated to Family 1): a-24-year-old woman was referred for epigastralgia. EGD revealed FGPs localized in the proximal stomach. Pathological biopsy results showed severe dysplasia and adenocarcinoma in situ. Her father was simultaneously diagnosed with FGPs with GC localized in the proximal stomach. We performed laparoscopic total gastrectomies with D1+lymph node dissection. They had the same gene mutation as the family 1. Here, we report two Asian families with GAPPS successfully treated via laparoscopic total gastrectomy.


The Development of Metaphor Comprehension and Its Relationship with Relational Verbal Reasoning and Executive Function.
Our main objective was to analyse the different contributions of relational verbal reasoning (analogical and class inclusion) and executive functioning to metaphor comprehension across development. We postulated that both relational reasoning and executive functioning should predict individual and developmental differences. However, executive functioning would become increasingly involved when metaphor comprehension is highly demanding, either because of the metaphors' high difficulty (relatively novel metaphors in the absence of a context) or because of the individual's special processing difficulties, such as low levels of reading experience or low semantic knowledge. Three groups of participants, 11-year-olds, 15-year-olds and young adults, were assessed in different relational verbal reasoning tasks-analogical and class-inclusion-and in executive functioning tasks-updating information in working memory, inhibition, and shifting. The results revealed clear progress in metaphor comprehension between ages 11 and 15 and between ages 15 and 21. However, the importance of executive function in metaphor comprehension was evident by age 15 and was restricted to updating information in working memory and cognitive inhibition. Participants seemed to use two different strategies to interpret metaphors: relational verbal reasoning and executive functioning. This was clearly shown when comparing the performance of the "more efficient" participants in metaphor interpretation with that of the "less efficient" ones. Whereas in the first case none of the executive variables or those associated with relational verbal reasoning were significantly related to metaphor comprehension, in the latter case, both groups of variables had a clear predictor effect.


Primary Tumors of the Posterior Pituitary Gland: A Systematic Review of the Literature in Light of the New 2017 World Health Organization Classification of Pituitary Tumors.
The rare clinical entity of primary posterior pituitary tumors (PPTs) includes pituicytomas, granular cell tumors, spine cell oncocytomas, and sellar ependymomas. The recent World Health Organization classification of PPTs based on thyroid transcription factor 1 positivity has led to more investigations into the epidemiology, clinical presentation, nature history, histologic features, and operative characteristics of these tumors. The aim of this review is to summarize the characteristics of primary PPTs. Our summary involved an in-depth review of the literature on PPTs. Our systematic review was carried out using the PubMed database and PRISMA guidelines. An initial search identified 282 publications. After strict application of the inclusion criteria, we found 16 articles for case series of patients with primary PPT (N > 5), which were included in our table for literature review. An additional 10 articles were review articles on PPTs published in the last 20 years and were used as resource for our systematic review. An extensive analysis was then performed to extract relevant clinical data with respect to the clinical radiologic histopathologic profile of primary PPTs and their treatment outcome. Primary PPTs are a rare group of pituicyte-derived low-grade nonneuroendocrine neoplasms that arise from the sellar region. The nondescript radiographic findings and subtle endocrine abnormalities also veil their accurate diagnostic prediction. As shown through the narrative as well as the literature review, there is still a lot to be understood about PPTs. A prospective multicenter registry of these rare tumors would benefit both the neurosurgical as well as the endocrinologic knowledge base.


Does routine patch angioplasty after carotid endarterectomy lessen the risk of perioperative stroke?
From 1964 through 1991 we performed primary closure of the arteriotomy in 1173 patients and patch angioplasty in 506 patients after carotid endarterectomy. The decision to patch was made at the surgeon's discretion. In general a patch was used for small arteries. In the primary closure group 32 patients (2.7%) had a perioperative stroke and in the patch angioplasty group 17 (3.4%) had a stroke. The difference (2.7% vs. 3.4%) was not significant (p < 0.5275, Fisher's exact two-tailed test). A total of 240 arteries were closed with a vein patch and 11 (4.6%) of these patients had a stroke; 266 were closed with a synthetic patch (Dacron, 211; polytetrafluoroethylene, 55) and six of the patients had a stroke (2.3%). The difference in stroke rate between the vein and synthetic patch groups (4.6% vs. 2.3%) was not significant (p < 0.2159). Patch angioplasty cannot be shown to reduce the incidence of perioperative stroke. Late carotid patency was not studied. This study supports a policy of selective patch angioplasty based on arterial size rather than patching all carotid arteries. When a patch is used, we prefer filamentous Dacron as the patch material.


Can birth weight predict later body composition in anorexia nervosa?
The relationship between birth weight and body composition at later stages in life was not studied previously in anorexia nervosa (AN). The aim of the following brief report is to present results concerning the relationship between birth weight and later body composition specifically in AN, and to check if the programming of body composition from birth weight is still detected in severely emaciated AN patients. One hundred and fifty-one female AN patients aged between 13 and 44 were recruited from 11 inpatient treatment facilities in France. Birth weight, body weight and height were obtained. Body composition was measured using bioelectrical impedance. Birth weight was significantly correlated to lifetime maximum body mass index (BMI; r=0.211, P=0.009) and significantly correlated to fat-free mass index (r=0.190, P=0.027) but not to fat mass index (FMI). This report confirms that even in AN when patients are severely emaciated and where fat-free mass (FFM) and fat mass (FM) are low, a link between birth weight and FFM and BMI can still be identified, independently from age. Further studies are needed on larger samples exploring other factors, such as gender, puberty and ethnicity.


RNA toxicity in human disease and animal models: from the uncovering of a new mechanism to the development of promising therapies.
Mutant ribonucleic acid (RNA) molecules can be toxic to the cell, causing human disease through trans-acting dominant mechanisms. RNA toxicity was first described in myotonic dystrophy type 1, a multisystemic disorder caused by the abnormal expansion of a non-coding trinucleotide repeat sequence. The development of multiple and complementary animal models of disease has greatly contributed to clarifying the complex disease pathways mediated by toxic RNA molecules. RNA toxicity is not limited to myotonic dystrophy and spreads to an increasing number of human conditions, which share some unifying pathogenic events mediated by toxic RNA accumulation and disruption of RNA-binding proteins. The remarkable progress in the dissection of disease pathobiology resulted in the rational design of molecular therapies, which have been successfully tested in animal models. Toxic RNA diseases, and in particular myotonic dystrophy, clearly illustrate the critical contribution of animal models of disease in translational research: from gene mutation to disease mechanisms, and ultimately to therapy development. This article is part of a Special Issue entitled: Animal Models of Disease.


Estrogenic in vitro evaluation of zearalenone and its phase I and II metabolites in combination with soy isoflavones.
Humans and animals are exposed to multiple substances in their food and feed that might have a negative health impact. Among these substances, the Fusarium mycoestrogen zearalenone (ZEN) and its metabolites α-zearalenol (α-ZEL) and α-zearalanol (α-ZAL) are known to possess endocrine disruptive properties. In a mixed diet or especially animal feed, these potential contaminants might be ingested together with naturally occurring phytoestrogens such as soy isoflavones. So far, risk assessment of potential endocrine disruptors is usually based on adverse effects of single compounds whereas studies investigating combinatorial effects are scarce. In the present study, we investigated the estrogenic potential of mycoestrogens and the isoflavones genistein (GEN), daidzein (DAI) and glycitein (GLY) as well as equol (EQ), the gut microbial metabolite of DAI, in vitro alone or in combination, using the alkaline phosphatase (ALP) assay in Ishikawa cells. In the case of mycoestrogens, the tested concentration range included 0.001 to 10 nM with multiplication steps of 10 in between, while for the isoflavones 1000 times higher concentrations were investigated. For the individual substances the following order of estrogenicity was obtained: α-ZEL > α-ZAL > ZEN > GEN > EQ > DAI > GLY. Most combinations of isoflavones with mycoestrogens enhanced the estrogenic response in the investigated concentrations. Especially lower concentrations of ZEN, α-ZEL and α-ZAL (0.001-0.01 nM) in combination with low concentrations of GEN, DAI and EQ (0.001-0.1 µM) strongly increased the estrogenic response compared to the single substances.


The association of alpha-synuclein with membranes affects bilayer structure, stability, and fibril formation.
The aggregation of alpha-synuclein is believed to be a critical factor in the etiology of Parkinson's disease. alpha-Synuclein is an abundant neuronal protein of unknown function, which is enriched in the presynaptic terminals of neurons. Although alpha-synuclein is found predominantly in the cytosolic fractions, membrane-bound alpha-synuclein has been suggested to play an important role in fibril formation. The effects of alpha-synuclein on lipid bilayers of different compositions were determined using fluorescent environment-specific probes located at various depths. alpha-Synuclein-membrane interactions were found to affect both protein and membrane properties. Our results indicate that in addition to electrostatic interactions, hydrophobic interactions are important in the association of the protein with the bilayer, and lead to disruption of the membrane. The latter was observed by atomic force microscopy and fluorescent dye leakage from vesicles. The kinetics of alpha-synuclein fibril formation were significantly affected by the protein association and subsequent membrane disruption, and reflected the conformation of alpha-synuclein. The ability of alpha-synuclein to disrupt membranes correlated with the binding affinity of alpha-synuclein for the particular membrane composition, and to the induced helical conformation of alpha-synuclein. Protofibrillar or fibrillar alpha-synuclein caused a much more rapid destruction of the membrane than soluble monomeric alpha-synuclein, indicating that protofibrils (oligomers) or fibrils are likely to be significantly neurotoxic.


Design, Synthesis, Characterization and Computational Studies of Mannich Bases Oxadiazole Derivatives as New Class of Jack Bean Urease Inhibitors.
Mannich bases consisting of 1,3,4-oxadiazole-2-thione (3 a-3 l) bearing various substituents were synthesized and found potent jack bean urease inhibitors. The prepared compounds showed significantly good inhibitory activities with IC50 values from 9.45±0.05 to 267.42±0.23 μM. The compound 3 k containing 4-chlorophenyl (-R) and 4-hydroxyphenyl (-R') was most active with IC50 9.45±0.05 μM followed by 3 e (IC50 22.52±0.15 μM) in which -R was phenyl and -R' was isopropyl group. However, when both -R and -R' were either 4-chlorophenyl groups (3 l) or only -R' was 4-nitrophenyl (3 i), both compounds were found inactive. The detailed binding affinities of the produced compounds with protein were explored through molecular docking and data-supported in-vitro enzyme inhibition profiles. Drug likeness was confirmed by in silico ADME investigations and molecular orbital analysis (HOMO-LUMO) and electrostatic potential maps were got from DFT calculations. ESP maps exposed that there are two potential binding sites with the most positive and most negative parts.


Combined topical-peribulbar anesthesia for cataract surgery.
To evaluate the anesthetic effect of single-point low-volume peribulbar anesthesia supplemented by topical anesthesia. Private ambulatory ophthalmic practice. Five hundred consecutive patients received 4 cc of lidocaine 2% with 200 units hyaluronidase as 1-point peribulbar anesthesia. This was supplemented by lidocaine 4%, 1 drop every 3 to 5 minutes for 3 instillations. Phacoemulsification and intraocular lens implantation were performed through a scleral tunnel or clear corneal approach. All patients were evaluated for intraoperative akinesia, lid closure, and anesthesia. One hour after surgery, the eye patch was removed and patients were evaluated for pain, discomfort, foreign-body sensation, diplopia, and lid closure. All patients had no pain to mild discomfort during surgery; 34% had total and 58% partial akinesia; 78% had poor orbicularis action (lid closure); 12% had subconjunctival hemorrhage. Postoperatively, 42% of patients had foreign-body sensation caused by conjunctival coaptation by diathermy or corneal edema. Diplopia occurred in 32% of patients but resolved within 1 hour after eye-patch removal, and partial ptosis occurred in 58%, resolving within 2 hours of patch removal. All patients had normal lid closure when the eye patch was removed. Low-volume 1-point peribulbar anesthesia supplemented by topical anesthesia was safe and effective and provided early visual recovery. Topical therapy can be started 1 hour postoperatively.


Experimental design for optimal parameter estimation of an enzyme kinetic process based on the analysis of the Fisher information matrix.
The investigation of enzyme kinetics is increasingly important, especially for finding active substances and understanding intracellular behaviors. Therefore, the determination of an enzyme's kinetic parameters is crucial. For this a systematic experimental design procedure is necessary to avoid wasting time and resources. The parameter estimation error of a Michaelis-Menten enzyme kinetic process is analysed analytically to reduce the search area as well as numerically to specify the optimum for parameter estimation. From analytical analysis of the Fisher information matrix the fact is obtained, that an enzyme feed will not improve the estimation process, but substrate feeding is favorable with small volume flow. Unconstrained and constrained process conditions are considered. If substrate fed-batch process design is used instead of pure batch experiments the improvements of the Cramer-Rao lower bound of the variance of parameter estimation error reduces to 82% for mu(max) and to 60% for K(m) of the batch values in average.


A survey exploring the practices of smoking cessation support among hospital-based healthcare providers.
Hospital visits constitute a 'window of opportunity' for initiating smoking cessation attempts, and healthcare providers (HCPs) play an important role in supporting patients to stop smoking. Yet, the current practices of supporting smoking cessation in the hospital setting are largely unexplored. The aim of this study was to explore practices of smoking cessation support among hospital-based HCPs. HCPs working in a large hospital in the secondary care sector completed an online, cross-sectional survey, including sociodemographic and work-related factors as well as 21 questions assessing practices of smoking cessation support based on the "five As" framework. Descriptive statistics were computed, and predictors of HCPs giving patients advice to stop smoking were explored using logistic regression analysis. All employees (N = 3998) in the hospital received a survey link; 1645 (41.1%) HCPs with daily patient contact completed the survey. Smoking cessation support in the hospital setting was limited with regard to assessment of smoking; providing information and advice; planning and referral for further support; and follow-up on smoking cessation attempts. Almost half (44.8%) of participating HCPs with daily patient contact never or rarely advise their patients to stop smoking. Physicians were more likely than nurses to advice patients to stop smoking, and HCPs in outpatient clinics were more likely to give advice than inpatient clinic HCPs. Smoking cessation support is very limited in the hospital-based healthcare setting. This is problematic, as hospital visits can be windows of opportunity to help patients change their health behaviour. An intensified focus on the implementation of hospital-based smoking cessation support is needed.


Ontogenetic shifts within the selfish herd: predation risk and foraging trade-offs change with age in colonial web-building spiders.
We examine costs and benefits associated with spatial position relative to spider age (size) in colonial web-building spiders. Predator attack and capture rates vary with position in the colony, and suggest that risk is higher for the smallest and the largest spiders on the periphery, and lower in the central core of the colony. Foraging success is greater on the periphery for small and medium spiders but does not differ significantly with position for larger spiders. Decreased predation risk may be the reason why larger spiders aggressively seek and defend positions in the colony core, demonstrating a "selfish herd effect" (Rayor and Uetz, 1990). Smaller (immature) spiders, unable to compete for protected web positions in the core, must trade-off potentially higher risk of predation to take advantage of higher prey availability on the periphery. Increased foraging success on the periphery may allow juvenile spiders to achieve the larger size necessary to compete successfully for protected core positions as adults. Spatial variation in size-related fitness trade-offs between predation risk and foraging success may explain why colonies are dynamic entities - with individual spiders exhibiting ontogenetic shifts in web location as they grow larger and mature-accounting for the characteristic age (size) structure ofMetepeira incrassata colonies.


Effect of television viewing on social-emotional competence of young Thai children.
Exposure time, program content and cultural context may affect the impact of television (TV) on the social-emotional competence (SEC) of children. This study examined the effects of TV viewing on the SEC of Thai infants. The study was based on a Thai birth cohort study from which duration and content of TV viewing and data from the Modified Infant-Toddler Social and Emotional Assessment instrument at 1 and 3 years of age were available. Generalized estimating equations were used to examine whether scores below the 10th national percentile were associated with TV viewing duration. The relationship between viewing duration and SEC risk was quadratic rather than linear. Viewing duration of 30-120 min/day was associated with a decreased risk of low overall SEC compared to non-viewers after adjustments for confounding factors. However, the beneficial effect diminished when the duration exceeded 120 min/day. Viewing educational programs was associated with a risk reduction of having low overall SEC compared to non-educational programs. These results suggest that a short period of TV viewing may be beneficial for the SEC of Thai infants, especially if the programs are educational.


Alcohol dehydrogenase gene expression in potato following elicitor and stress treatment.
A cDNA clone corresponding to a mRNA that rapidly accumulates during the hypersensitive-like response induced by elicitor treatment of potato (Solanum tuberosum L.) tuber was characterized. The clone encodes a polypeptide (Mr = 41,097) having 83%-85% amino acid identity with known plant alcohol dehydrogenase sequences (ADH; EC 188.8.131.52). The identity of the clone was confirmed by measuring the ADH enzyme activity in extracts of Escherichia coli transformed with the cDNA clone. In potato tuber disks, a wide range of stresses, including treatment with fatty acid elicitors, salicylic acid, UV light and anaerobiosis, was shown to induce accumulation of Adh transcripts. In stems, a high constitutive level of Adh transcripts could be detected in 4-week old plants, but not in 8-week old plants. However, the mRNA could be induced to accumulate in stems of 8-week old plants by treatment with arachidonic acid elicitor or by anaerobiosis. Induction in leaves was also obtained during anaerobiosis and after treatment with a Phytophthora infestans mycelial homogenate.


Variants in DNA double-strand break repair and DNA damage-response genes and susceptibility to lung and head and neck cancers.
Cigarette smoking is the major risk factor for lung cancer, and together with alcohol for head and neck (H--N) cancer. These genotoxics produced DNA damage and particularly double-strand breaks (DSB) that are removed by various repair pathways. To understand the initiation of these cancers, we performed a genotype analysis to correlate some variants in specific genes in a case-control study of lung and H-N cancers. In a discovery phase, we sequenced DNA samples of 32 healthy Caucasians to describe genetic variants in 30 genes involved in the repair of DSB and in DNA damage response. 625 variants were detected on 29 out of the 30 genes successfully screened by sequencing exons, parts of introns and flanking regions. These included 470 non-exonic variants, from which 33 insertions/deletions, and 155 exonic alterations, corresponding to 59 non synonymous polymorphisms. 223 of these variants were not previously described. In total, 379 variants were successfully genotyped in a case-control study restricted to smokers including 151 lung cases, 251 H-N cases, and 172 controls. To account for multiple testing, we associated to each p-value a proportion of false positives (q-value). Haplotype-analysis suggested potential associations (p < 0.05) between lung cancer and 2 genes (RECQL4 and RAD52), which came with q-value of 8%, and between H-N cancer and 1 gene (DNA-PK) but with q-value of 56%. The 3 genes are key players for regulating the efficiency of DSB repair. Large-scale studies are needed to show if any of these 3 variants are truly associated with an increased risk of cancer.


Transmission/disequilibrium tests of androgen receptor and glutathione S-transferase pi variants in prostate cancer families.
Population-based case-control studies have found relationships between risk of prostate cancer and genetic polymorphisms in the CAG repeat and GGC repeat of the X-linked androgen receptor gene (AR) as well as the autosomal gene coding for glutathione S-transferase pi (GSTP1). This family-based study utilized the transmission disequilibrium test to examine whether there was evidence that these polymorphisms could account for familial aggregation of prostate cancer. Seventy-nine North American pedigrees were studied. Most of these families had 3 or more affected first-degree relatives. Genotype information was obtained on 578 individuals. The reconstruction combined transmission disequilibrium test (RC-TDT) was used to test for linkage. There was no evidence of linkage to the CAG and GGC repeat sequences in the AR gene or the pentanucleotide (ATAAA) repeat in the GSTP1 gene when each allele was analyzed separately or when alleles were grouped by repeat length. Our findings do not support the hypothesis that familial clustering of prostate cancer in high-risk families is attributable to these genetic variants.


Hyaluronic acid stimulates protein kinase activity in intact cells and in an isolated protein complex.
The addition of hyaluronate to intact chick embryonic heart fibroblasts enriched with a hyaluronate-binding protein (HABP) stimulated phosphorylation of tyrosine and serine/threonine residues in cellular proteins. A protein complex containing a hyaluronate-binding protein (cell-HABP) was isolated from the cultured heart fibroblasts. The isolated complex (Mr approximately 1 x 10(6] contained phosphoproteins that exhibited protein kinase activity specifically stimulated by hyaluronate. Both tyrosine and serine residues in the protein complex were phosphorylated in response to this glycosaminoglycan. The hyaluronate-stimulated protein kinase activity was tightly associated with cell-HABP in vitro; enzyme activity co-immunoprecipitated with cell-HABP using a monospecific anti-HABP antibody and co-eluted with cell-HABP when chromatographed on a column of Sephacryl S-1000 in 2.0 M guanidine hydrochloride. The uniqueness of the cell-HABP-associated protein kinase activity was suggested by both its specific response to hyaluronate, relative to related glycosaminoglycans such as heparin and chondroitin sulfate or to growth factors such as epidermal growth factor or insulin, and its antigenic distinction from other protein kinases such as growth factor receptors. These results point to a new mechanism by which glycosaminoglycans, such as hyaluronate, may modify cell behavior.


Indian hedgehog and syndecans-3 coregulate chondrocyte proliferation and function during chick limb skeletogenesis.
Hedgehog proteins exert critical roles in embryogenesis and require heparan sulfate proteoglycans (HS-PGs) for action. Indian hedgehog (Ihh) is produced by prehypertrophic chondrocytes in developing long bones and regulates chondrocyte proliferation and other events, but it is not known whether it requires HS-PGs for function. Because the HS-PG syndecan-3 is preferentially expressed by proliferating chondrocytes, we tested whether it mediates Ihh action. Primary chick chondrocyte cultures were treated with recombinant Ihh (rIhh-N) in absence or presence of heparinase I or syndecan-3 neutralizing antibodies. While rIhh-N stimulated proliferation in control cultures, it failed to do so in heparinase- or antibody-treated cultures. In reciprocal gain-of-function studies, chondrocytes were made to overexpress syndecan-3 by an RCAS viral vector. Cells became more responsive to rIhh-N, but even this response was counteracted by heparinase or antibody treatment. To complement the in vitro data, RCAS viral particles were microinjected in day 4-5 chick wing buds and effects of syndecan-3 misexpression were monitored over time. Syndecan-3 misexpression led to widespread chondrocyte proliferation and, interestingly, broader expression and distribution of Ihh. In addition, the syndecan-3 misexpressing skeletal elements were short, remained cartilaginous, lacked osteogenesis, and exhibited a markedly reduced expression of collagen X and osteopontin, products characteristic of hypertrophic chondrocytes and bone cells. The data are the first to indicate that Ihh action in chondrocyte proliferation involves syndecan-3 and to identify a specific member of the syndecan family as mediator of hedgehog function.


Comparative Study of Two Infrared Meibographers in Evaporative Dry Eye Versus Nondry Eye Patients.
To compare the measurement of the meibomian gland (MG) dropout between two infrared meibographers in patients with and without dry eye. The right eyelids of each patient were imaged using the Antares and Cobra meibography devices. All images were analyzed using Phoenix software to calculate the percentage of the MG dropout. Lipid layer thickness, eyelid margin characteristics, ocular surface staining, MG secretion, number of expressible glands, and noninvasive tear breakup time were also evaluated. A comparison between nondry eye and evaporative dry eye was performed. Eighty participants (mean age, 36.93 years and 51.3% women) were included, of which 67.5% had nondry eye. A significant difference was observed in the dropout percentage of the superior eyelid between the Antares and Cobra devices (P=0.007) for all participants and when only the nondry eye patients were examined. In patients with dry eye, no significant differences were found in the dropout measurements of both eyelids. Statistically significant differences in the MG dropout percentage in the upper eyelid of nondry eye patients were obtained from both meibographers. The measurements were similar in patients with dry eye, suggesting that the two instruments can be interchanged.


Blood glycoprotein from antarctic fish. Possible conformational origin of antifreeze activity.
High resolution 1H NMR and circular dichroism (CD) measurements have been performed on aqueous solutions of antarctic fish antifreeze glycoprotein. The carbohydrate contribution ot the observed CD spectrum has been estimated from closely analogous model compounds. The residual peptide contribution cannot be interpreted of the known spectral behaviour of alpha-helix, beta-sheet and random coil. Instead it resembles the CD spectrum of beta-structure in position, magnitude and spectral form, but is of opposite sign, indicating a specific but unusual peptide conformation, which we suggest may be stabilised by non-bonded interactions between the peptide backbone and the carbohydrate sidechains. Previous evidence which supports this interpretation is reviewed. NMR and CD measurements between -2 and +30 degrees C are consistent with conformational stability throughout the biologically relevant temperature range. The mechanism of the antifreeze activity is discussed in terms of the spatial and orientational correlations of sugar hydroxy groups and water in the liquid and solid states. The implication of an ordered peptide structure is explained by the comparison of the antifreeze glycoprotein with synthetic water-soluble polymers which also exhibit limited antifreeze properties.


Systemic 8-OH-DPAT challenge causes hyperventilation largely via activating pre-botzinger complex 5-HT1A receptors.
Systemic 8-OH-DPAT (a 5-HT1A receptor agonist) challenge evokes hyperventilation independent of peripheral 5-HT1A receptors. Though the pre-Botzinger Complex (PBC) is critical in generating respiratory rhythm and activation of local 5-HT1A receptors induces tachypnea via disinhibition of local GABAA neurons, its role in the respiratory response to systemic 8-OH-DPAT challenge is still unclear. In anesthetized rats, 8-OH-DPAT (100 μg/kg, iv) was injected twice to confirm the reproducibility of the evoked responses. The same challenges were performed after bilateral microinjections of (S)-WAY-100135 (a 5-HT1A receptor antagonist) or gabazine (a GABAA receptor antagonist) into the PBC. Our results showed that: 1) 8-OH-DPAT caused reproducible hyperventilation associated with hypotension and bradycardia; 2) microinjections of (S)-WAY-100135 into the PBC attenuated the hyperventilation by ˜60 % without effect on the evoked hypotension and bradycardia; and 3) the same hyperventilatory attenuation was also observed after microinjections of gabazine into the PBC. Our data suggest that PBC 5-HT1A receptors play a key role in the respiratory response to systemic 8-OH-DPAT challenge likely via disinhibiting local GABAergic neurons.


Expression of E-cadherin, alpha-catenin, and beta-catenin in the process of lymph node metastasis in oral squamous cell carcinoma.
Regional lymph node metastasis is a very important prognostic indicator. In the metastatic process, reduction in cell to cell adhesion including E-cadherin-catenin cell adhesion complex is an essential step. We investigated immunohistochemical expression of E-cadherin, alpha-catenin and beta-catenin in 159 tissue samples from patients with oral squamous cell carcinoma and examined the correlation between their expressions and the presence of regional lymph node metastasis. Significantly greater reduction in expression levels of E-cadherin, alpha-catenin and beta-catenin was found in the metastatic group (n=64) compared to the nonmetastatic group (n=95) (P=0.007, 0.001, 0.001, respectively). However, there was no significant correlation between their expressions and the features of the regional metastasis, the number of metastatic lymph nodes or the presence of extracapsular metastasis. These data suggest that evaluation of the immunohistochemical expression of E-cadherin, alpha-catenin and beta-catenin is extremely valuable for the diagnosis of metastatic occurrence.


SARS-CoV-2 infection and stem cells: Interaction and intervention.
The emergence of the novel severe acute respiratory coronavirus 2 (SARS-CoV-2) in China and its rapid national and international spread have created a global health emergency. The resemblance with SARS-CoV in spike protein suggests that SARS-CoV-2 employs spike-driven entry into angiotensin-converting enzyme 2 (ACE2)-expressing cells. From a stem cell perspective, this review focuses on the possible involvement of ACE2+ stem/progenitor cells from both the upper and lower respiratory tracts in coronavirus infection. Viral infection-associated acute respiratory distress syndrome and acute lung injury occur because of dysregulation of the immune response. Mesenchymal stem cells appear to be a promising cell therapy given that they favorably modulate the immune response to reduce lung injury. The use of exogenous stem cells may lead to lung repair. Therefore, intervention by transplantation of exogenous stem cells may be required to replace, repair, remodel, and regenerate lung tissue in survivors infected with coronavirus. Ultimately, vaccines, natural killer cells and induced-pluripotent stem cell-derived virus-specific cytotoxic T lymphocytes may offer off-the-shelf therapeutics for preventing coronavirus reemergence.


Middle cerebral artery flow velocity decreases and electroencephalogram (EEG) changes occur as acute hypercapnia reverses.
To define the changes in middle cerebral artery flow velocity (Vmca) and the electroencephalogram (EEG) during rapid reduction in arterial carbon dioxide (PCO2) from acute hypercapnia. DESIGN. Human volunteer study. University-affiliated hospital experimental laboratory. Nine healthy volunteers aged 25-35 years. Subjects rebreathed exhaled gas from a bag pre-filled with 5% carbon dioxide (CO2) in oxygen, up to an end-tidal CO2 of 10% or to the limit of discomfort, when they were disconnected. Middle cerebral artery blood flow velocity was continuously measured by transcranial Doppler ultrasound, quantitative EEG was recorded and hemodynamics were monitored non-invasively. Vmca closely correlated with end-tidal CO2 changes (r=0.65, p<0.001). When rebreathing ceased, there was a rapid decline in end-tidal CO2 and in Vmca to baseline within 42+/-14 s, followed by a rapid further decline in both variables to below baseline. End-tidal CO2 reached a nadir of 4.4+/-1.1% at 146+/-79 s, with Vmca decreasing to 37+/-10 cm/s at 104+/-65 s (a 40% reduction and 31% below basal values). Electroencephalogram alpha activity significantly decreased and delta activity increased during hypercapnia. During subsequent hypocapnia, delta activity decreased back to baseline, while alpha activity increased, but remained below baseline. These findings demonstrate the rapid changes occurring in cerebral blood flow during rapid declines in arterial CO2 and the consequent potential for producing brain ischemia and hemodynamic alterations if inadvertent hyperventilation occurs following institution of mechanical ventilation for acute hypercapnia.


Effect of charged groups on the adsorption and penetration of proteins onto and into carboxymethylated poly(HEMA) hydrogels.
A range of carboxymethylated poly(hydroxyethyl methacrylate) (CM-PHEMA) hydrogels with varying degrees of carboxymethylation was synthesized for a systematic study of the effects of ionized groups ('charge') on the uptake by hydrogel matrices of the proteins, lysozyme and human serum albumin (HSA). Using a radiolabel-tracer technique, X-ray photoelectron spectroscopy, and laser scanning confocal microscopy, we attempted to differentiate between protein molecules that were irreversibly adsorbed onto the hydrogel surface and those that penetrated into the hydrogel matrix. The effective pore size of the CM-PHEMA hydrogels was modelled and compared with the known molecular dimensions of the two proteins. The effects of the presence of varying amounts of ionized groups in the hydrogel matrix differed for the two proteins. For lysozyme, increased uptake was observed at higher carboxymethylation; this is interpreted as resulting from a combination of electrostatic attraction and increasing ease of penetration of the protein into the more porous hydrogel matrix. For HSA, on the other hand, the uptake was primarily by surface adsorption, with little diffusive penetration into the matrix.


Brain afferents to the lateral caudal ventrolateral medulla: a retrograde and anterograde tracing study in the rat.
The ventrolateral medulla (VLM) modulates autonomic functions, motor reactions and pain responses. The lateralmost part of the caudal VLM (VLMlat) was recently shown to be the VLM area responsible for pain modulation. In the present study, the brain sources of VLMlat afferent fibers were determined by tract-tracing techniques. Following injection of cholera toxin subunit B into the VLMlat, retrogradely labeled neurons in the forebrain occurred at the somatosensory, insular, motor, limbic and infralimbic cortices, and at the central amygdaloid nucleus. Retrogradely labeled neurons in diencephalic regions were observed in the lateral hypothalamus, posterior hypothalamus and paraventricular nucleus. In the brainstem, retrograde labeling occurred at the periaqueductal gray, red nucleus, parabrachial area, nucleus raphe magnus, nucleus tractus solitarii, lateral reticular nucleus and dorsal and ventral medullary reticular formation. In the cerebellum, retrogradely labeled neurons occurred at the lateral nucleus. Following injections of the anterograde tracer biotinylated dextran amine (BDA) into the lateral hypothalamus or paraventricular nucleus, anterogradely labeled fibers were mainly observed in the VLMlat. Injections of BDA into the periaqueductal gray, red nucleus or lateral nucleus of the cerebellum resulted in anterograde labeling in the VLMlat and lateral reticular nucleus. The present study gives an account of the brain regions putatively involved in triggering the modulatory actions elicited from the VLMlat. These include areas committed to somatosensory processing, autonomic control, somatic and visceral motor activity and affective reactions. The findings suggest that the VLMlat may play a major homeostatic role in the integration of nociception with other brain functions.


Preliminary analysis of the mouse cerebellum proteome.
This paper reports on the initial analysis of protein expression in the mouse cerebellum with the proteomics approach. Proteins from cerebellar tissue homogenates were separated by two-dimensional gel electrophoresis, and the proteins were stained with colloidal Coomassie Blue to produce a high-resolution map of the cerebellum proteome. Selected proteins from this map were digested with trypsin, and the resulting tryptic peptides were analyzed by matrix-assisted laser desorption/ionization mass spectrometry and liquid chromatography-electrospray quadrupole ion trap mass spectrometry. The mass spectrometric data were used to identify the proteins through searches of the SWISSPROT protein sequence database. To date, 30 prominent proteins with various functional characteristics were identified. These data will be used for future studies of differential protein expression in mouse models of neurological disorders.


In vivo imaging reveals rapid astrocyte depletion and axon damage in a model of neuromyelitis optica-related pathology.
Neuromyelitis optica (NMO) is an autoimmune disease of the central nervous system, which resembles multiple sclerosis (MS). NMO differs from MS, however, in the distribution and histology of neuroinflammatory lesions and shows a more aggressive clinical course. Moreover, the majority of NMO patients carry immunoglobulin G autoantibodies against aquaporin-4 (AQP4), an astrocytic water channel. Antibodies against AQP4 can damage astrocytes by complement, but NMO histopathology also shows demyelination, and - importantly-axon injury, which may determine permanent deficits following NMO relapses. The dynamics of astrocyte injury in NMO and the mechanisms by which toxicity spreads to axons are not understood. Here, we establish in vivo imaging of the spinal cord, one of the main sites of NMO pathology, as a powerful tool to study the formation of experimental NMO-related lesions caused by human AQP4 antibodies in mice. We found that human AQP4 antibodies caused acute astrocyte depletion with initial oligodendrocyte survival. Within 2 hours of antibody application, we observed secondary axon injury in the form of progressive swellings. Astrocyte toxicity and axon damage were dependent on AQP4 antibody titer and complement, specifically C1q. In vivo imaging of the spinal cord reveals the swift development of NMO-related acute axon injury after AQP4 antibody-mediated astrocyte depletion. This approach will be useful in studying the mechanisms underlying the spread of NMO pathology beyond astrocytes, as well as in evaluating potential neuroprotective interventions. Ann Neurol 2016;79:794-805.