Adaptation of the Sensititre broth microdilution technique to antimicrobial susceptibility testing of Mycoplasma hyopneumoniae.
A broth microdilution technique is described for determining the antimicrobial susceptibility of Mycoplasma hyopneumoniae, using commercially prepared Sensititre plates. Twenty-five field isolates and two reference strains (J & 232), were tested against seven antimicrobials. Field isolates were tested in duplicate and reference strains, four times to estimate reproducibility. Ninety-seven percent of the duplicate MIC results for the field isolates were in agreement, or within one log2 dilution. Similar results were obtained with the reference strains. The isolates were susceptible to lincomycin-spectinomycin, tylosin and oxytetracycline or resistant to amoxycillin, apramycin and erythromycin. Susceptibility to furaltadone varied. This method retains the accuracy and reproducibility of broth MIC determinations, while avoiding the lengthy preparation of antimicrobial dilutions normally associated with more traditional methods.


Scalene lymph node biopsy in the preoperative evaluation of patients with recurrent cervical cancer.
From 1982 to 1987, 24 patients with recurrent carcinoma of the cervix underwent scalene lymph node biopsy prior to exploration for exenteration. Patients with palpable nodes were excluded from the study. There was no significant morbidity associated with the procedure. None of the 24 patients was found to have metastases to the scalene lymph nodes. From this study it would seem unjustifiable to perform this procedure on all patients undergoing evaluation for exenteration.


Lymphocyte beta 2-adrenoceptors and plasma catecholamines in patients with cirrhosis.
The hemodynamic response to propranolol in patients with cirrhosis is heterogeneous. In some patients, there is an expected decrease in portal pressure, whereas in others, portal pressure fails to decrease despite adequate beta-blockade. It has been suggested that this lack of response could be related to down-regulation of beta 2-adrenoceptors, promoted by the increased adrenergic activity frequently found in decompensated cirrhosis. The present study investigated this hypothesis by measuring the density and affinity of lymphocyte beta 2-adrenoceptors (L-beta 2-AR), an established index of beta 2-adrenoceptors in target organs, and the plasma levels of norepinephrine and epinephrine in a group of 32 patients with cirrhosis and portal hypertension. The portal pressure response to propranolol administration (0.1 mg/kg + 2 mg/h) was also investigated (n = 27). Patients with cirrhosis had increased norepinephrine levels (605 +/- 360 vs. 224 +/- 110 pg/mL in controls; P less than 0.001), but plasma epinephrine level was not increased (136 +/- 72 vs. 111 +/- 22 pg/mL in controls; NS). There were no differences between cirrhotic patients and controls in the density of L-beta 2-AR (1398 +/- 489 vs. 1278 +/- 356 receptors/cell; NS). Portal pressure decreased greater than 10% in 12 patients (responders) and less than 10% in the remaining 15 (nonresponders). Contrary to previous suggestions, there were no significant differences between responders and nonresponders in relation to the density of L-beta 2-AR (1362 +/- 527 vs. 1487 +/- 419 receptors/cell; NS), to the affinity of these receptors (0.15 +/- 0.27 vs. 0.13 +/- 0.12 nmol/L; NS), to the plasma levels of norepinephrine (661 +/- 508 vs. 621 +/- 256 pg/mL; NS), and to plasma epinephrine concentration (141 +/- 90 vs. 140 +/- 75 pg/mL; NS). These results show that the response of portal pressure to propranolol administration is neither related to the density and affinity of L-beta 2-AR nor to the plasma levels of norepinephrine and epinephrine. Thus, the determination of these parameters cannot substitute measurements of portal pressure to identify those patients with an adequate portal pressure response to propranolol treatment.


Targeting colorectal cancer via its microenvironment by inhibiting IGF-1 receptor-insulin receptor substrate and STAT3 signaling.
The tumor microenvironment (TME) exerts critical pro-tumorigenic effects through cytokines and growth factors that support cancer cell proliferation, survival, motility and invasion. Insulin-like growth factor-1 (IGF-1) and signal transducer and activator of transcription 3 (STAT3) stimulate colorectal cancer development and progression via cell autonomous and microenvironmental effects. Using a unique inhibitor, NT157, which targets both IGF-1 receptor (IGF-1R) and STAT3, we show that these pathways regulate many TME functions associated with sporadic colonic tumorigenesis in CPC-APC mice, in which cancer development is driven by loss of the Apc tumor suppressor gene. NT157 causes a substantial reduction in tumor burden by affecting cancer cells, cancer-associated fibroblasts (CAF) and myeloid cells. Decreased cancer cell proliferation and increased apoptosis were accompanied by inhibition of CAF activation and decreased inflammation. Furthermore, NT157 inhibited expression of pro-tumorigenic cytokines, chemokines and growth factors, including IL-6, IL-11 and IL-23 as well as CCL2, CCL5, CXCL7, CXCL5, ICAM1 and TGFβ; decreased cancer cell migratory activity and reduced their proliferation in the liver. NT157 represents a new class of anti-cancer drugs that affect both the malignant cell and its supportive microenvironment.


Formulation and delivery issues for monoclonal antibody therapeutics.
Antibodies can have exquisite specificity of target recognition and thus generate highly selective outcomes following their systemic administration. While antibodies can have high specificity, the doses required to treat patients, particularly for a chronic condition, are typically large. Fortunately, advances in production and purification capacities have allowed for the exceptionally large amounts of highly purified monoclonal antibodies to be produced. Additionally, genetic engineering of antibodies has provided a stable of antibody-like proteins that can be easier to prepare. Together, these advances have made antibody-based therapies one of the most commonly pursued pharmaceuticals in biotechnology pipelines. With this success, however, has come a series of technical challenges in the formulation of antibody-based materials to maintain sufficient stability in a variety of configurations and sometimes at particularly high concentrations. This review focuses on issues related to identifying and verifying stable antibody-based formulations.


Exploring staff willingness to attend work during a disaster: a study of nurses employed in four Australian emergency departments.
Much of the literature about emergency nurses willingness to work during disasters has been from a non-Australian perspective. Despite the many recent disasters, little is known of Australian nurse's willingness to participate in disaster response. This paper presents findings from a study that explored nurses willingness to attend work during a disaster and the factors that influenced this decision. Data were collected consecutively using a combination of focus group and interview methods. Participants in this study, registered nurses from emergency departments, were recruited through convenience sampling from four hospitals in Australia. Participant narrative was electronically recorded, transcribed and thematically analysed. The participants for both the focus groups and interviews compromised a mix of ages, genders and years of experience as emergency nurses from across four jurisdictions within Australia. Three major themes that influenced willingness emerged with a number of subthemes. Theme one reflected the uncertainty of the situation such as the type of disaster. The second theme surrounded the preparedness of the workplace, emergency nurse and colleagues, and the third theme considered personal and professional choice based on home and work circumstances and responsibilities. The decision to attend work or not during a disaster, includes a number of complex personal, work-related and professional factors that can change, depending on the type of disaster, preparedness of the work environment and the emergency nurses' personal responsibilities at that time.


Spontaneous resolution of solitary osteochondroma in the young adult.
Spontaneous resolution of a solitary osteochondroma is rare. Such a case is presented in a patient nearing skeletal maturity. Based on a search of the English literature this is the first such report in a patient of this age.


Linking Annual Prescription Volume of Antidepressants to Corresponding Web Search Query Data: A Possible Proxy for Medical Prescription Behavior?
Persons using the Internet to retrieve medical information generate large amounts of health-related data, which are increasingly used in modern health sciences. We analyzed the relation between annual prescription volumes (APVs) of several antidepressants with marketing approval in Germany and corresponding web search query data generated in Google to test whether web search query volume may be a proxy for medical prescription practice. We obtained APVs of several antidepressants related to corresponding prescriptions at the expense of the statutory health insurance in Germany from 2004 to 2013. Web search query data generated in Germany and related to defined search terms (active substance or brand name) were obtained with Google Trends. We calculated correlations (Person's r) between the APVs of each substance and the respective annual "search share" values; coefficients of determination (R) were computed to determine the amount of variability shared by the 2 variables. Significant and strong correlations between substance-specific APVs and corresponding annual query volumes were found for each substance during the observational interval: agomelatine (r = 0.968, R = 0.932, P = 0.01), bupropion (r = 0.962, R = 0.925, P = 0.01), citalopram (r = 0.970, R = 0.941, P = 0.01), escitalopram (r = 0.824, R = 0.682, P = 0.01), fluoxetine (r = 0.885, R = 0.783, P = 0.01), paroxetine (r = 0.801, R = 0.641, P = 0.01), and sertraline (r = 0.880, R = 0.689, P = 0.01). Although the used data did not allow to perform an analysis with a higher temporal resolution (quarters, months), our results suggest that web search query volume may be a proxy for corresponding prescription behavior. However, further studies analyzing other pharmacologic agents and prescription data that facilitate an increased temporal resolution are needed to confirm this hypothesis.


Morphologic and Molecular Characteristics of Bladder Cancer.
Bladder cancer is the fourth most common cancer in men, and is associated with significant morbidity and mortality. Pathologic evaluation of urothelial cancers relies predominantly on histomorphologic features but can be aided in a small subset of cases by immunohistochemical analyses. Distinction of papillary versus flat lesions, low-grade versus high-grade cytology, and histologic variants and the presence or absence of invasive tumor is important for proper clinical management. Advances in the molecular alterations associated with the various subtypes of urothelial carcinoma have been made but such studies are ongoing.


Esters and amides of 2,3-dimethoxy-8,9-methylenedioxy-benzo[i]phenanthridine-12-carboxylic acid: potent cytotoxic and topoisomerase I-targeting agents.
The exceptional topoisomerase I-targeting activity and antitumor activity of 5-(2-N,N-dimethylamino)ethyl-8,9-dimethoxy-2,3-methylenedioxy-5H-dibenzo[c,h][1,6]naphthyridin-6-one (ARC-111, topovale) prompted studies on similarly substituted benzo[i]phenanthridine-12-carboxylic ester and amide derivatives. Among the benzo[i]phenanthridine-12-carboxylic esters evaluated, the 2-(N,N-dimethylamino)ethyl, 2-(N,N-dimethylamino)-1-methylethyl, and 2-(N,N-dimethylamino)-1,1-dimethylethyl esters possessed similar cytotoxicity, ranging from 30 to 55 nM in RPMI8402 and KB3-1 cells. Several of the carboxamide derivatives possess potent topoisomerase I-targeting activity and cytotoxicity. The 2-(N,N-dimethylamino)ethyl, 2-(N,N-diethylamino)ethyl, and 2-(pyrrolidin-1-yl)ethyl amides were among the more cytotoxic benzo[i]phenanthridine-12-carboxylic derivatives, with IC50 values ranging from 0.4 to 5.0 nM in RPMI8402 and KB3-1 cells.


An improved methodology to detect human T cell receptor beta variable family gene expression patterns.
Comprehensive gene expression analysis of the T cell receptor repertoire of an individual can be very useful in evaluating the immune response in a variety of conditions. Antibody-based analysis methods can detect approximately 60% of the human T cell receptor beta variable (TCRBV) proteins, while gene expression analysis, primarily through employment of the polymerase chain reaction (PCR), has had somewhat greater success in the detection of additional TCRBV families. Many of these previous PCR methods, however, have been unable to detect all 91 alleles of the human TCRBV genes. This is primarily due to either deficiencies in the amplification of all of the variable beta families, subfamilies, and alleles, or the prior lack of a systematic classification of the TCR variable family gene segment sequences. We describe here a real-time reverse transcription polymerase chain reaction-based method, which allows efficient automation and integration of amplification, detection, and analysis with sequence-specific detection of all T cell receptor beta variable gene families, subfamilies, and alleles. This method, which in itself contributes significant improvements over existing technologies through its comprehensiveness and efficiency, also functions independently of variables such as sample source and sample processing and has the ability to run on multiple real-time PCR platforms, affording one the implementation of personal preferences.


Gendered sexual uses of alcohol and associated risks: a qualitative study of Nigerian University students.
Alcohol misuse among young people is a global phenomenon. In many countries, young people engage in heavy drinking and this exacerbates risky sexual behaviour. In Nigeria, alcohol held multiple roles in the traditional era but was mainly consumed by adult males for pleasure. Adult females and young people were culturally constrained from drinking in most communities. In contemporary Nigeria, young people's drinking is increasing, and many engage in sexual intercourse under the influence of alcohol. This study draws on the traditional gender and social sexual scripts to explore the factors that motivate young people to use alcohol for sexual purposes. In-depth interviews were conducted with 19 to 23-year old male and female undergraduate students from a Nigerian university. Thematic analysis was conducted with the aid of NVivo 10 software. Men drink to become confident to initiate sexual relationships, stimulate sexual urges, prolong erection, increase sexual satisfaction and become more aggressive during sexual intercourse. Women also drink to be bold in initiating sexual relationships, for sexual arousal and to increase satisfaction. Relatedly, not every brand of alcohol is used for sexual purposes. For example, while men use 'herbal' alcoholic beverages and a mixture of locally-produced gin and marijuana, women use champagne and other flavoured alcoholic beverages. The results also revealed that young people use alcohol or salt in a bid to prevent conception after sexual intercourse. Adherence to the traditional gender (masculinity) and social sexual scripts amongst men and the enactment of what appears to be a new form of femininity script amongst women contribute to a culturally specific understanding of the motivations to use alcohol for sexual purposes. Evidence-based strategies should be employed to distribute information about the consequences of sexual intercourse under the influence of alcohol.


Anaphylactic reactions to modified fluid gelatins.
Use of modified fluid gelatins as a plasma expander is of interest in human clinical medicine due to osmotic pressure similarities with plasma proteins. However, adverse reactions such as urticaria, edema, and/or anaphylactic shock occur and can lead to diagnostic problems. In addition, mechanisms of these reactions are poorly understood. We report three cases of anaphylactic shock studied by skin tests and, for the first time, in vitro leukocyte histamine release (LHR). Intradermal skin tests were significantly positive for concentrations of 1:1000 to 1:10 of the commercial preparation used before the reaction in each patient. Thirty control subjects were negative even for the undiluted preparation. Positive LHR was obtained from patients' leukocytes washed and then incubated in Tris-albumin Ca++ Mg++ buffer with serial dilutions of fluid gelatins; controls were negative. Addition of D2O (50%) caused significant increase of LHR in patients but had no effect on controls. In conclusion, skin tests and LHR might be valuable in diagnosis of patients reactive to gelatins. Furthermore, these findings suggest release of mediators from mast cells or basophils, but discrimination between immunologic and idiosyncratic pharmacologic mechanism was not obtained.


A precise review on NAATs-based diagnostic assays for COVID-19: A motion in fast POC molecular tests.
Diagnosis is one of the main strategies to deal with infectious and deadly diseases such as coronavirus disease 2019 (COVID-19). The global pandemic of COVID-19 has led to an immediate need to expand rapid diagnostic techniques. New isothermal-based methods are being developed for COVID-19 detection aiming to resolve the limitations related to the reverse transcription-quantitative polymerase chain reaction (RT-qPCR) method through immediate samples processing and minimizing false-negative or ambiguous results. Advances in nucleic acid amplification techniques (NAATs) can provide affordable and easy-to-use diagnostic platforms with high sensitivity and specificity in order to be available to the public as approved commercial kits. The development of point-of-care (POC) testing can assist in rapid clinical decision-making and mitigate burdens on health care facilities. Finally, we discussed the different diagnostic methods based on NAATs for COVID-19 in detail. Comparative parameters are addressed for all assays and Emergency Use Authorizations (EUA)-approved commercial tests are cited. Isothermal-coupled methods and LAMP-based molecular methods have been suggested as suitable portable tests with high diagnostic speed for use in POC testing.


In-hospital recurrence and functional outcome between ischemic stroke caused by intracranial arterial dissection and intracranial atherosclerosis: Retrospective cohort study of the nationwide multicenter registry.
Intracranial arterial dissection (ICAD) and intracranial atherosclerotic stenosis (ICAS) are often difficult to differentiate, and studies on their background factors and prognosis are scarce. Information on prognosis, including recurrence, is necessary for stroke care, and clarification of epidemiological and clinical differences between the two diseases is important for appropriately handling their heterogeneity. This study aimed to determine the association of ICAD and ICAS with in-hospital recurrence and prognosis and compare their background and clinical findings. In this multicenter cohort study, we retrospectively analyzed data from the Saiseikai Stroke Database. Adults with ischemic stroke caused by ICAD or ICAS were included in this study. Patients' backgrounds and clinical findings were compared between the ICAD and ICAS groups. The outcome showed an association of ICAD with in-hospital recurrence of ischemic stroke and poor functional outcome relative to ICAS. Multivariable logistic regression analyses were performed to calculate the adjusted odds ratios (ORs) for ICAD with 95% confidence intervals (CIs) for each outcome. Among 15,622 patients registered in the Saiseikai Stroke Database, 2,020 were enrolled (ICAD group: 89; ICAS group: 1,931). In the ICAD group, 65.2% of the patients were aged <64 years. Vascular lesion location was more common in ICAD with the vertebral artery [42 (47.2%)], anterior cerebral artery [20 (22.5%)], and middle cerebral artery (MCA) [16 (18.0%)], and in ICAS with MCA 1046 (52.3%). Multivariable logistic regression analyses of the association between ICAD and in-hospital recurrence and poor functional outcome yielded a crude OR (95% CI) of 3.26 (1.06-9.97) and 0.97 (0.54-1.74), respectively, relative to ICAS. ICAD was associated with a higher in-hospital recurrence than ICAS; however, there was no significant difference in prognosis between the two groups. Differences in background characteristics and vessel lesions may be of interest in these two diseases.


Characterization of differences in insulin receptors from young and old red blood cells.
It has been demonstrated that young RBCs (reticulocytes and early mature erythrocytes) possess more insulin receptors than old RBCs (late mature erythrocytes) but it is not yet known whether insulin receptors on young and old RBCs are regulated similarly. In the present investigation insulin receptors on young and old RBCs have, therefore, been studied in five normal male subjects before and after 2 days dexamethasone ingestion (0.5 mg tablet every 6 h) and, in the same subjects, before and 5 h after ingestion of 75 g glucose. The results obtained clearly demonstrate that dexamethasone increases insulin receptor concentration while glucose ingestion increases both insulin receptor affinity and concentration on young RBCs. By contrast, neither stimuli modify insulin receptors on old RBCs. Studies on RBCs are usually performed on the whole RBC population not taking into account this differential responsiveness of receptors on young versus old RBCs; consequently, this phenomenon might be responsible of the fact that some data reported on RBCs are not in agreement with those reported on monocytes or adipocytes and it should be taken into consideration when using RBCs to evaluate insulin receptor regulation.


Colloidal gold submonolayer-coated thin-film glass plates for waveguide-coupled surface plasmon resonance sensors.
A novel optical sensor based on spectroscopic measurement of the plasmon absorption of a colloidal gold submonolayer immobilized upon a planar waveguide has been developed for label-free detection of biomolecular interactions at the sensor's surface. We fabricated the sensor by locally modifying a 50-microm-thick glass plate with hemoglobin (Hb) and then self-assembling gold colloids from the aqueous solution onto the Hb-modified area of the glass plate. Polychromatic light from a xenon-arc lamp was launched into the thin-film glass plate by use of a broadband fiber-coupling method. With the use of a CCD detector to monitor the light beam emitted from an end face of the glass plate, the plasmon absorption spectrum of the colloidal gold submonolayer was determined to depend on the polarization states of the guided light and to change with the refractive index of the medium in contact with the colloids. In addition to simplicity of fabrication and the ease of use, the sensor yields a larger absorbance sensitivity than the normal transmission measurement.


Quantifying chromosomal instability from intratumoral karyotype diversity using agent-based modeling and Bayesian inference.
Chromosomal instability (CIN)-persistent chromosome gain or loss through abnormal mitotic segregation-is a hallmark of cancer that drives aneuploidy. Intrinsic chromosome mis-segregation rate, a measure of CIN, can inform prognosis and is a promising biomarker for response to anti-microtubule agents. However, existing methodologies to measure this rate are labor intensive, indirect, and confounded by selection against aneuploid cells, which reduces observable diversity. We developed a framework to measure CIN, accounting for karyotype selection, using simulations with various levels of CIN and models of selection. To identify the model parameters that best fit karyotype data from single-cell sequencing, we used approximate Bayesian computation to infer mis-segregation rates and karyotype selection. Experimental validation confirmed the extensive chromosome mis-segregation rates caused by the chemotherapy paclitaxel (18.5 ± 0.5/division). Extending this approach to clinical samples revealed that inferred rates fell within direct observations of cancer cell lines. This work provides the necessary framework to quantify CIN in human tumors and develop it as a predictive biomarker.


Molecular dynamics in thin grafted and spin-coated polymer layers.
Broadband Dielectric Spectroscopy is employed to study the molecular dynamics in thin layers of grafted and spin-coated polydimethylsiloxane (PDMS) and of the type-A polymer cis-1,4-polyisoprene (PI). Strong resemblance with low-molecular-weight systems is found but additionally the conformation of the chain and its extension becomes important. For PI a confinement-induced relaxation is found. It is observable if the layer thickness is comparable to the end-to-end vector of the chain and it depends strongly on the separation between interfaces. In contrast, the dynamic (and hence the calorimetric) glass transition is not affected by the geometrical confinement.


Feasibility and safety of percutaneous laser revascularization using the Biosense system in porcine hearts.
Direct myocardial revascularization (DMR) is being explored to improve symptoms in patients with refractory ischemic coronary syndromes. This study assessed the safety and feasibility of percutaneous DMR using the Biosense non-fluoroscopic navigation system (incorporating the holmium : yttrium aluminium garnet laser) in pig hearts. Twenty pigs underwent left ventricular mapping using the Biosense system, followed by percutaneous DMR with a holmium : yttrium aluminium garnet laser at 2 J x 1 pulse, and were sacrificed acutely (n = 5), at 24 h (n = 5), and at 3 weeks (n = 10) after the DMR procedure. The hearts were examined grossly and microscopically. Serial creatine kinase-MB level was measured up to 24 h. There were no procedural complications. Animals received 13 +/- 2 channels in 19 +/- 7 min. The catheter was able to reach all regions, including the septum. There was no increase in creatine kinase-MB level up to 24 h. One animal died within an hour of the procedure. There was one sealed perforation into the subepicardial fat as a result of same site laser activation. Histologic evaluation in the acute and 24 h groups revealed laser channels which were 3.6 +/- 2.2 mm long, 1.5 +/- 0.7 mm wide, with an entry angle of 73 +/- 12 degrees. Channels were filled with platelet thrombi acutely and were surrounded by well-defined rims of thermal coagulation and an 'impact zone' of viable myocardium; at 3 weeks, no channels remained patent and they had been replaced by well-healed granulation tissue. It is feasible and safe to perform percutaneous DMR with the Biosense system and the channels created with the chosen laser parameters are rapidly sealed with platelet thrombi and at 3 weeks are replaced by well-healed granulation tissue.


A mixed-methods study of the management of hearing loss associated with otitis media with effusion in children with Down syndrome.
To scope current service provision across England for management of otitis media with effusion and hearing loss in children with Down syndrome; to explore professional decision-making about managing otitis media with effusion and hearing loss; and to explore patient and public views on the direction of future research. Mixed methods including a service evaluation of NHS clinical practice through a structured telephone survey; a qualitative study of professional decision-making with in-depth interviews collected and analysed using grounded theory methods; patient/public involvement consultations. Twenty-one audiology services in England took part in the evaluation; 10 professionals participated in the qualitative study; 21 family members, 10 adults with Down syndrome and representatives from two charities contributed to the consultations. There was variation across services in the frequency of routine hearing surveillance, approaches to managing conductive hearing loss in infancy and provision of hearing aids and grommets. There was variation in how professionals describe their decision-making, reflecting individual treatment preferences, differing approaches to professional remit and institutional factors. The consultations identified that research should focus on improving practical support for managing the condition and supporting decision-making about interventions. There is system-level variation in the provision of services and individual-level variation in how professionals make clinical decisions. As a consequence, there is inequity of access to hearing health care for children with Down syndrome. Future research should focus on developing core outcomes for research and care, and on improving decision support for families.


Identification of cattle carrying alleles associated with resistance and susceptibility to the Bovine Leukemia Virus progression by real-time PCR.
Previous studies have shown a significant association between polymorphisms of the BoLA DRB3 gene and Bovine Leukemia Virus (BLV) infection profile. The presence of allele *1501 has been associated with high proviral load in peripheral blood while allele *0902 has been associated with low proviral load. The purpose of this study was to develop allele-specific real-time PCRs to identify cattle carrying alleles associated with resistance (BoLA DRB3*0902) or susceptibility (BoLA DRB3*1501) to the BLV progression. Specific primers were designed and differential amplification was carried out by real-time PCR and monitored by SYBR® Green dye in DNA samples from peripheral blood. Conditions were also adjusted for traditional PCR amplification (end point amplification). These methods are rapid, simple and suitable for high throughput screening, and could aid in marker-assisted selection of BLV-resistant and susceptible cattle.


Subunit structure of the purified human placental insulin receptor. Intramolecular subunit dissociation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
Insulin receptors purified from human placental membranes by gel-filtration and insulin-agarose affinity chromatography were found to be composed of eight different high molecular weight complexes as identified by nonreducing sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. The subunit stoichiometry of these different high molecular weight forms of the insulin receptor were determined by comparisons of silver-stained gel profiles with the autoradiograms of 125I-insulin specifically cross-linked to the alpha subunit and [gamma-32P]ATP specifically autophosphorylated beta subunit gel profiles. Two-dimensional SDS-polyacrylamide gel electrophoresis in the absence and presence of reductant confirmed the subunit stoichiometries as alpha 2 beta 2, alpha 2 beta beta 1, alpha 2 (beta 1)2, alpha 2 beta, alpha 2 beta 1, alpha 2, alpha beta, and beta, where alpha is the Mr = 130,000 subunit, beta is the Mr = 95,000 subunit, and beta 1 is the Mr = 45,000 subunit. Treatment of the insulin receptor preparations with oxidized glutathione or N-ethylmaleimide prior to SDS-polyacrylamide gel electrophoresis increased the relative amount of the alpha 2 beta 2 complex concomitant with a total disappearance of the alpha 2 beta, alpha 2 beta 1, alpha 2, and free beta forms. The effects of oxidized glutathione were found to be completely reversible upon extensive washing of the treated insulin receptors. In contrast, the effects of N-ethylmaleimide were totally irreversible by washing, consistent with known sulfhydryl alkylating properties of this reagent. The formation of these lower molecular weight insulin receptor subunit complexes was further demonstrated to be due to SDS/heat-dependent intramolecular sulfhydryl-disulfide exchange occurring within the alpha 2 beta 2 complex. These studies demonstrate that the largest disulfide-linked complex (alpha 2 beta 2) is the predominant insulin receptor form purified from the human placenta with the other complexes being generated by proteolysis and by internal subunit dissociation.


Meta-analysis of the accuracy of p16 or p16/Ki-67 immunocytochemistry versus HPV testing for the detection of CIN2+/CIN3+ in triage of women with minor abnormal cytology.
Women with atypical squamous cells of undetermined significance (ASC-US) can be triaged accurately with a high-risk human papillomavirus (hrHPV) test to identify those who need a referral. However, the triage of low-grade squamous intraepithelial lesion (LSIL) with hrHPV testing has very low specificity. Overexpression of p16, with or without Ki-67, indicates neoplastic transformation of human papillomavirus-infected cervical cells and may more accurately predict underlying cervical intraepithelial neoplasia of grade 3 or worse (CIN3+). A literature search was conducted in 3 bibliographic databases. Studies were selected if they included women with ASC-US or LSIL who were triaged with dual staining (p16/Ki-67) and/or p16 staining and, if available, with a comparator hrHPV test to detect cervical intraepithelial neoplasia of grade 2 or worse (CIN2+) or CIN3+. Thirty-eight studies were eligible. The sensitivity of p16 staining for CIN3+ was significantly lower than that of hrHPV DNA testing (ratio for ASC-US, 0.87; 95% confidence interval [CI], 0.78-0.97; ratio for LSIL, 0.86; 95% CI, 0.80-0.93). In contrast, the specificity of p16 staining was substantially higher with relative specificities of 1.60 (95% CI, 1.35-1.88) and 2.29 (95% CI, 2.05-2.56) for ASC-US and LSIL respectively. Dual staining was as sensitive as hrHPV DNA testing but was more specific (ratio for ASC-US, 1.65; 95% CI, 1.42-1.92; ratio for LSIL, 2.45; 95% CI, 2.17-2.77). This meta-analysis confirms that p16 staining and p16/Ki-67 staining are more specific for CIN2+/CIN3+ than hrHPV DNA testing. Although p16 staining is less sensitive for CIN3+ than hrHPV DNA testing, dual staining has similar sensitivity.


Prothrombinex-induced thrombosis and its management with regional plasminogen activator in hepatic failure.
Fulminant hepatic failure causes a bleeding diathesis as a result of impaired synthesis of hepatic clotting factors, thrombocytopenia, fibrinolysis and disseminated intravascular coagulation. Administration of clotting factor concentrates can cause thrombosis in patients with acute hepatic failure. Regional infusion of recombinant tissue-type plasminogen activator may be used to induce local thrombolysis. A case report of a five-year-old child is presented and the literature is reviewed.


Temporal trends of cause-specific mortality after diagnosis of atrial fibrillation.
Reports of outcomes after atrial fibrillation (AF) diagnosis are conflicting. The aim of this study was to investigate mortality and hospitalization rates following AF diagnosis over time, by cause and by patient features. Individuals aged ≥16 years with a first diagnosis of AF were identified from the UK Clinical Practice Research Datalink-GOLD dataset from 1 January 2001, to 31 December 2017. The primary outcomes were all-cause and cause-specific mortality and hospitalization at 1 year following diagnosis. Poisson regression was used to calculate rate ratios (RRs) for mortality and incidence RRs (IRRs) for hospitalization and 95% confidence intervals (CIs) comparing 2001/02 and 2016/17, adjusted for age, sex, region, socio-economic status, and 18 major comorbidities. Of 72 412 participants, mean (standard deviation) age was 75.6 (12.4) years, and 44 762 (61.8%) had ≥3 comorbidities. All-cause mortality declined (RR 2016/17 vs. 2001/02 0.72; 95% CI 0.65-0.80), with large declines for cardiovascular (RR 0.46; 95% CI 0.37-0.58) and cerebrovascular mortality (RR 0.41; 95% CI 0.29-0.60) but not for non-cardio/cerebrovascular causes of death (RR 0.91; 95% CI 0.80-1.04). In 2016/17, deaths caused from dementia (67, 8.0%), outstripped deaths from acute myocardial infarction, heart failure, and acute stroke combined (56, 6.7%, P < .001). Overall hospitalization rates increased (IRR 2016/17 vs. 2001/02 1.17; 95% CI, 1.13-1.22), especially for non-cardio/cerebrovascular causes (IRR 1.42; 95% CI 1.39-1.45). Older, more deprived, and hospital-diagnosed AF patients experienced higher event rates. After AF diagnosis, cardio/cerebrovascular mortality and hospitalization has declined, whilst hospitalization for non-cardio/cerebrovascular disease has increased.


Studies of the growth and infiltration of experimental tumor of the pancreas in rabbits.
An experimentally transplanted pancreatic cancer was induced by means of an intraductal injection of Vx2 carcinoma supernatant into the pancreatic duct of rabbits. The presence of ductal obstruction appeared to assist the nidation and growth of injected cancer cells in the pancreas. Tumors induced in the head of the pancreas grew by extension to the body and tail where they also produced interspersed, nodular tumor masses. Tumors induced in the body and tail region, on the other hand, grew by direct expansion towards the head, but this did not produce the interspersed nodular tumors in the head portion. These observations indicate that cancer of the head of the pancreas associated with the stenosis or obstruction of the pancreatic duct resulted in ductal dissemination and extensive lymphatic infiltration in the body and tail, a wider cancer invasion than recognizable at the gross cancer border. This extension was noncontinuous and interspersed, so that radical curative surgery for such cancer of the head of the pancreas would indicate total pancreatectomy.


Attenuation of neonatal ischemic brain damage using a 20-HETE synthesis inhibitor.
20-Hydroxyeicosatetraenoic acid (20-HETE) is a cytochrome P450 metabolite of arachidonic acid that that contributes to infarct size following focal cerebral ischemia. However, little is known about the role of 20-HETE in global cerebral ischemia or neonatal hypoxia-ischemia (H-I). The present study examined the effects of blockade of the synthesis of 20-HETE with N-hydroxy-N'-(4-n-butyl-2-methylphenyl) formamidine (HET0016) in neonatal piglets after H-I to determine if it protects highly vulnerable striatal neurons. Administration of HET0016 after H-I improved early neurological recovery and protected neurons in putamen after 4 days of recovery. HET0016 had no significant effect on cerebral blood flow. cytochrome P450 4A immunoreactivity was detected in putamen neurons, and direct infusion of 20-HETE in the putamen increased phosphorylation of Na(+), K(+) -ATPase and NMDA receptor NR1 subunit selectively at protein kinase C-sensitive sites but not at protein kinase A-sensitive sites. HET0016 selectively inhibited the H-I induced phosphorylation at these same sites at 3 h of recovery and improved Na(+), K(+) -ATPase activity. At 3 h, HET0016 also suppressed H-I induced extracellular signal-regulated kinase 1/2 activation and protein markers of nitrosative and oxidative stress. Thus, 20-HETE can exert direct effects on key proteins involved in neuronal excitotoxicity in vivo and contributes to neurodegeneration after global cerebral ischemia in immature brain.


"Oh yes, oh yes, these are the atoms!" A personal recollection from the times of the invention of the STM.
The scanning tunneling microscope, invented and developed in the 1980s in the IBM research laboratory in Rüschlikon, has become the dominent scientific tool in surface science and together with its younger brother the atomic force microscope, is widely used also in many other modern research areas. This account contains very personal anecdotal memories from a colleague who worked in a neighboring lab in Rüschlikon describing some events at the periphery of the development of this ingenious instrument.


Assessment of the formation of inorganic oxidation by-products during the electrocatalytic treatment of ammonium from landfill leachates.
This work investigates the formation of oxidation by-products during the electrochemical removal of ammonium using BDD electrodes from wastewaters containing chlorides. The influence of the initial chloride concentration has been experimentally analyzed first, working with model solutions with variable ammonium concentration and second, with municipal landfill leachates. Two different levels of chloride concentration were studied, i) low chloride concentrations ranging between 0 and 2000 mg/L and, ii) high chloride concentrations ranging between 5000 and 20,000 mg/L. Ammonium removal took place mainly via indirect oxidation leading to the formation of nitrogen gas and nitrate as the main oxidation products; at high chloride concentration the formation of nitrogen gas and the rate of ammonium removal were both favored. However, chloride was also oxidized during the electrochemical treatment leading to the formation of free chlorine responsible of the ammonium oxidation, together with undesirable products such as chloramines, chlorate and perchlorate. Chloramines appeared during the treatment but they reached a maximum and then started decreasing, being totally removed when high chloride concentrations were used. With regard to the formation of chlorate and perchlorate once again the concentration of chloride exerted a strong influence on the formation kinetics of the oxidation by-products and whereas at low chloride concentrations, chlorate appeared like an intermediate compound leading to the formation of perchlorate, at high chloride concentrations chlorate formation was delayed significantly and perchlorate was not detected during the experimental time. Thus this work contributes first to the knowledge of the potential hazards of applying the electro-oxidation technology as an environmental technology to deal with ammonium oxidation under the presence of chloride and second it reports efficient conditions that minimize or even avoid the formation of undesirable by-products.


Role of age on transmitral flow velocity patterns in assessing left ventricular diastolic function in normal infants and children.
This study demonstrated that both peak E and flow velocity integral of early diastole increased to reach the older children's values by 36 months of age and leveled off thereafter, whereas both peak A and flow velocity integral of atrial contraction had little change. These results suggest that age-related changes in E wave reflect the maturational or developmental alterations in LV diastolic properties, especially in the relaxation process.


PEI - a potent, but not harmless, mucosal immuno-stimulator of mixed T-helper cell response and FasL-mediated cell death in mice.
Polyethyleneimine (PEI) is one of the most effective gene delivery systems available today. However, very little is known about its ability to stimulate a systemic immune response and the molecular mechanisms thereof. However, this information is vital for the future development of new gene delivery systems. Here we address this issue by studying gene expression profiles from spleen lymphocytes after in vivo immunization of mice with PEI formulated with a reporter plasmid (PEI+) or the formulation alone (PEI-). PEI- was found to provoke the activation of genes with important immunostimulatory functions, but without the necessary costimulatory signals. PEI+ resulted in: a mixed Th1/Th2 response; activation of both CD8(+) and CD4(+) T cells, with a larger effect on CD4(+); and FasL-mediated antigen-induced cell death. A comparison of the immune responses of PEI+ with that of the clinically used tetanus toxoid-aluminum phosphate vaccine showed that the DNA vaccine provoked a stronger immune response as compared to the protein vaccine. However, many genes involved in other cellular responses such as apoptosis, stress responses and oncogenesis were activated in PEI+, supporting the theory of immunostimulation by danger genes, but also pointing toward possible adverse reactions such as Alzheimer's disease.


Comparison of byakujutsu (Atractylodes rhizome) and sojutsu (Atractylodes lancea rhizome) on anti-inflammatory and immunostimulative effects in vitro.
The Japanese Pharmacopoeia defines byakujutsu (Atractylodes rhizome) as the rhizome of Atractylodes japonica or A. macrocephala and sojutsu (Atractylodes lancea rhizome) as the rhizome of A. lancea, A. chinensis, or their interspecific hybrids. Because their pharmaceutical uses differ in traditional Japanese Kampo medicine and traditional Chinese medicine, with less apparent scientific evidence, we compared the pharmacological properties between byakujutsu and sojutsu. Crude drug specimens of byakujutsu (n = 40) and sojutsu (n = 49) obtained in markets were identified by their species using DNA profiling. Their pharmacological properties were evaluated by the inhibitory effect of a MeOH extract of the samples on nitric oxide (NO) production by lipopolysaccharide-stimulated murine macrophage-like RAW264.7 cells and by the inducing effect of boiling water extract of the samples on granulocyte-colony stimulating factor (G-CSF) secretion from murine normal colonic epithelial MCE301 cells. We authenticated A. macrocephala (n = 8), A. japonica (n = 35), and the hybrid between A. macrocephala and A. japonica (n = 1), and they were used as byakujutsu. We authenticated A. chinensis (n = 25), A. lancea (n = 14), and the hybrid between A. chinensis and A. lancea (n = 6), and they were used as sojutsu. The inhibitory effects of byakujutsu on NO production were significantly higher than those of sojutsu (P < 0.05). This activity of A. japonica rhizome was significantly higher than that of A. macrocephala rhizome and A. lancea rhizome (P < 0.01). The activity of A. chinensis rhizome was significantly higher than that of A. lancea rhizome (P < 0.05). The extract of A. japonica rhizome significantly induced G-CSF secretion from MCE301 cells in a concentration-dependent manner. These effects of byakujutsu samples were not significantly different from those of sojutsu samples. A. japonica rhizome had significantly higher activity than A. macrocephala rhizome; however, there were no statistically significant differences among A. japonica, A. chinensis, and A. lancea. The pharmacological differences of byakujutsu and sojutsu may not be large among highly variated crude drug samples with average values, and quality control with the identification of the original plant species of byakujutsu and sojutsu may guarantee their pharmacological properties.


Criminal characteristics and psychopathy in women in prison.
To compare the factors and facets of psychopathy based on criminal characteristics in a sample of women in prison. Descriptive, comparative, cross-sectional study composed of a sample of 41 women incarcerated in the Ambato prison in Ecuador. In an individual session, the Hare Psychopathy Scale Revised was applied. Recidivist women, with a juvenile criminal history and who are admitted to the maximum-security ward, have a higher score in the affective facet of the PCL-R. In addition, those women who are in the maximum-security pavilion have scored high in factor 2 (social deviance), mainly in the antisocial aspect. This subgroup of women in prison is characterized by lack of remorse, emotional insensitivity, manipulation, inability to accept responsibility for their own actions, and superficial affection. Further expansion of the study of psychopathy in women is required.


A simple procedure for preparing substrate for Pleurotus ostreatus cultivation.
The use of wooden crates for composting a mixture of 70% grass, (Digitaria decumbens), and 30% coffee pulp, combined with 2% Ca(OH)(2), was studied as a method for preparing substrate for the cultivation of Pleurotus ostreatus. Crate composting considerably modified the temperature pattern of the substrate in process, as compared to pile composting, where lower temperatures and less homogeneous distributions were observed. Biological efficiencies varied between 59.79% and 93% in the two harvests. Based on statistical analysis significant differences were observed between the treatments, composting times and in the interactions between these two factors. We concluded that it is possible to produce P. ostreatus on a lignocellulosic, non-composted, non-pasteurized substrate with an initial pH of 8.7, and that composting for two to three days improves the biological efficiency.


Vorinostat, a histone deacetylase inhibitor, facilitates fear extinction and enhances expression of the hippocampal NR2B-containing NMDA receptor gene.
Histone acetylation, which alters the compact chromatin structure and changes the accessibility of DNA to regulatory proteins, is emerging as a fundamental mechanism for regulating gene expression. Histone deacetylase (HDAC) inhibitors increase histone acetylation and enhance fear extinction. In this study, we examined whether vorinostat, an HDAC inhibitor, facilitates fear extinction, using a contextual fear conditioning (FC) paradigm, in Sprague-Dawley rats. We found that vorinostat facilitated fear extinction. Next, the levels of global acetylated histone H3 and H4 were measured by Western blotting. We also assessed the effect of vorinostat on the hippocampal levels of NMDA receptor mRNA by real-time quantitative PCR (RT-PCR) and protein by Western blotting. 2 h after vorinostat administration, the levels acetylated histones and NR2B mRNA, but not NR1 or NR2A mRNA, were elevated in the hippocampus. The NR2B protein level was elevated 4 h after vorinostat administration. Last, we investigated the levels of acetylated histones and phospho-CREB (p-CREB) binding at the promoter of the NR2B gene using the chromatin immunoprecipitation (ChIP) assay followed by RT-PCR. The ChIP assay revealed increases in the levels of acetylated histones and they were accompanied by enhanced binding of p-CREB to its binding site at the promoter of the NR2B gene 2 h after vorinostat administration. These findings suggest that vorinostat increases the expression of NR2B in the hippocampus by enhancing histone acetylation, and this process may be implicated in fear extinction.


The difference in myocardial injuries and mitochondrial damages between asphyxial and ventricular fibrillation cardiac arrests.
Ventricular fibrillation (VF) and asphyxia account for most cardiac arrests but differ in cardiac arrest course, neurologic deficit, and myocardial damage. In VF resuscitation, cardiac mitochondria were known to be damaged via excess generation of reactive oxygen species. This study evaluated the difference of cardiac mitochondrial damages between VF and asphyxial cardiac arrests. In the VF + electrical shock (ES) group, VF was induced and untreated for 5 minutes, followed by 1 minute of cardiopulmonary resuscitation (CPR) and 1 ES of 5 J. Animals were killed immediately after ES. In the asphyxia group, cardiac arrest was induced by airway obstruction, and then pulselessness was maintained for 5 minutes, followed by 1 minute of CPR. The animals were killed immediately after CPR. The histology and ultrastructural changes of myocardium and complex activities and respiration of mitochondria were evaluated. The mitochondrial permeability transition pore opening was measured based on mitochondrial swelling rate. The histopathologic examinations showed myocardial necrosis and mitochondrial damage in both cardiac arrests. Instead of regional damages of myocardium in the VF + ES group, the myocardial injury in the asphyxia group distributed diffusely. The asphyxia group demonstrated more severe mitochondrial damage than the VF + ES group, which had a faster mitochondrial swelling rate, more decreased cytochrome c oxidase activity, and more impaired respiration. Both VF and asphyxial cardiac arrests caused myocardial injuries and mitochondrial damages. Asphyxial cardiac arrest presented more diffuse myocardial injuries and more severe mitochondrial damages than VF cardiac arrest.


The utilization of an ultrasound-guided 8-gauge vacuum-assisted breast biopsy system as an innovative approach to accomplishing complete eradication of multiple bilateral breast fibroadenomas.
Ultrasound-guided vacuum-assisted breast biopsy technology is extremely useful for diagnostic biopsy of suspicious breast lesions and for attempted complete excision of appropriately selected presumed benign breast lesions. A female patient presented with 16 breast lesions (eight within each breast), documented on ultrasound and all presumed to be fibroadenomas. Over a ten and one-half month period of time, 14 of these 16 breast lesions were removed under ultrasound guidance during a total of 11 separate 8-gauge Mammotome(R) excision procedures performed during seven separate sessions. Additionally, two of these 16 breast lesions were removed by open surgical excision. A histopathologic diagnosis of fibroadenoma and/or fibroadenomatous changes was confirmed at all lesion excision sites. Interval follow-up ultrasound imaging revealed no evidence of a residual lesion at the site of any of the 16 original breast lesions. This report describes an innovative approach of utilizing ultrasound-guided 8-gauge vacuum-assisted breast biopsy technology for assisting in achieving complete eradication of multiple bilateral fibroadenomas in a patient who presented with 16 documented breast lesions. As such, this innovative approach is highly recommended in similar appropriately selected patients.


An additive regression model for investigating the relationship between childhood health and socio-economic status.
Health risks associated with socio-economic status (SES) have extensively been studied in epidemiology. It is not uncommon that data used exhibit spatial correlation, nonlinear effects, overdispersion and heterogeneity, and structured additive regression (STAR) models permit incorporating these features in a single analytical framework. Nevertheless, most STAR models assume constant spatial effects. However, due to social or disease transmission processes, covariates may be space-varying. We explore this feature by fitting a multinomial logistic model on a joint response variable constructed from four health indicators (i.e. a child having fever, diarrhoea, or being stunted and underweight) and examine its relationship with SES estimated as a space-varying coefficient (SVC) variable. Implementation of the model follow a Bayesian framework. Our comparison with models that assume constant spatial effects shows superiority of the SVC model as well as confirms the fact that SES varies in space.


Mannose dependent tightening of the rat ependymal cell barrier. In vivo and in vitro study using neoglycoproteins.
The possible role of carbohydrate binding proteins (lectins) and glycoconjugates in the formation of junctions ensuring tightening between ependymal cells was studied using synthetic glycoconjugates, the neoglycoproteins. These compounds are prepared by substituting bovine serum albumin with sugar residues and additional labelling (or not) with fluorescein or biotin. Injections of these components into the cerebral ventricles of adult rats resulted in a binding pattern which could be related to their carbohydrate composition. Mannose-containing neoglycoproteins were bound to ependymal cell cilia and penetrated rapidly the brain tissue. Such phenomenon was not seen with glucose- or galactose-containing neoglycoprotein molecules. In contrast, mannose-, galactose- and glucose-containing neoglycoproteins bound strongly to some endothelial cells around blood vessels. Fluorescent unglycosylated serum albumin did not bind to any brain structures. In contrast, co-injection of mannose-containing non-fluorescent neoglycoproteins with the other fluorescent compounds (including fluorescent sugar-free BSA) resulted in the penetration of the fluorescent compounds into the brain tissue. This internalization into brain was attributed to disaggregation of junctions between ependymal cells. Cultured ependymal cells behaved likewise. In short term experiments (5 min-1 h), only the mannose-containing neoglycoproteins bound strongly to the ependymal cells, particularly to the cilia. In long term experiments (1-9 days), mannose-containing neoglycoproteins specifically induced the disappearance of junctions between the cultured cells. These results emphasize the importance of mannose-dependent recognition system in the maintenance of junctions between ependymal cells, where a mannose-binding lectin has been previously detected.


Evaluation of different column chemistries for fast urinary metabolic profiling.
Fast analytical methodologies are mandatory for large scale metabolic profiling. Here, we present a thorough evaluation of different column chemistries in combination with different mobile phases for fast LC-MS urinary metabolic profiling. Three porous HILIC materials were investigated, next to core-shell C18-, XB-C18- and PFP-RPLC material. The performance of the selected column chemistries was tested in a non-targeted manner with pooled urine samples and in a targeted manner with a set of 54 common urinary metabolites. In order to evaluate the differential behaviour of the tested columns in a targeted manner, we applied a peak scoring algorithm. This algorithm takes into account several quality criteria such as retention time, dead time, peak height and peak shape. In general, HILIC columns generate more retention for polar metabolites. Our results show that the diol-HILIC column outperforms the RPLC columns. However, because of their opposite nature, comprehensive behaviour is observed as well, which was shown by investigating gender differences in a small urinary sample set. All applied column chemistries enabled sufficient peak capacity within a short gradient time.


Characterization of the increased biological potency in BALB/C 3T3 cells of two analogs of human insulinlike growth factor I which have reduced affinity for the 28 K cell-derived binding protein.
We have characterized the biological activity of two analogs of insulinlike growth factor I (IGF I) which have significantly reduced affinity for the soluble 28 K binding proteins which are secreted by various cell types. The analogs, which were made by site-directed mutagenesis of a synthetic gene encoding for IGF I, are [Gln 3, Ala 4, Tyr 15, Leu 16] IGF I and an analog in which the first 16 amino acids of IGF I were replaced with the first 17 amino acids of insulin (B-chain mutant). These two peptides have 100-fold and greater than 1,000-fold lower affinity, respectively, than IGF I for the 28 K binding protein present in the conditioned medium of two cell types, the clonal rat vascular smooth muscle line A10, and BALB/C 3T3 cells. The 28 K protein secreted by BALB/C 3T3 cells has fivefold-lower apparent affinity for both IGF I and [Gln 3, Ala 4, Tyr 15, Leu 16] IGF I than does the 28 K protein secreted by A 10 cells. Conditioned medium from these two cell types has similar amounts of unoccupied 28 K protein as evidenced by the ability of 125I-IGF I to specifically bind to and be covalently bound to the protein after treatment with the bifunctional cross-linking reagent disuccinimidyl suberate. In the presence of 0.1% calf serum, IGF I and [Gln 3, Ala 4, Tyr 15, Leu 16] IGF I stimulate DNA synthesis in A10 cells with ED50 = 0.4 nM, and in BALB/C 3T3 cells with ED50 = 10 nM and 1.3 nM, respectively. Thus, these peptides are equipotent in A10 cells, but the mutant peptide is ten times more active than IGF I in BALB/C 3T3 cells. A10 cells can be made ten times less sensitive to IGF I by performing the incubation in the presence of conditioned media from BALB/C 3T3 cells but not from A10 cells. The activity of [Gln 3, Ala 4, Tyr 15, Leu 16] IGF I is not altered under these conditions. Thus, the conditioned media, which contain 28 K proteins secreted by A10 cells and BALB/C 3T3 cells, have different effects on the biological action of IGF I. These data suggest that the 28 K binding proteins can have important effects on the sensitivity of tissues to IGF I and that the B-chain mutant and [Gln 3, Ala 4, Tyr 15, Leu 16] IGF I will be useful in assessing the biological role of these proteins.


Contrast Induced Sialadenitis with Low Volume Non-ionic Contrast in a Patient with Cerebral Venous Sinus Thrombosis.
Background: Contrast Induced Sialadenitis or Iodide Mumps refers to non-suppurative inflammation of salivary glands following intravenous iodinated contrast administration. It is a rare adverse effect of iodinated contrast with only a few cases reported worldwide. It is hypothesized to be an idiosyncratic reaction due to toxic accumulation of iodine in salivary glands. Case Report: We report a case of a 40-year-old female patient who underwent CECT brain after intravenous injection of 40ml of non-ionic iodinated contrast and developed symmetric painless swelling in bilateral submandibular triangles within five hours of contrast administration. Ultrasound with color doppler and MR imaging was performed which confirmed the diagnosis of contrast induced sialadenitis. Sialadenitis was managed conservatively and resolved slowly over eight days. Conclusion: Though it is a rare self-limiting adverse event of iodinated contrast, it must be known to the radiologist as well as the clinician to avoid unnecessary work up and manage the patient better. The online version contains supplementary material available at 10.1007/s12070-023-03575-x.


A systematic genotype and subgenotype re-ranking of hepatitis B virus under a novel classification standard.
It is commonly noticed that chaotic and inefficient subgenotyping are universally used academically and clinically, a standardized HBV genotype/subgenotype classification criterion is urgently acquired. Sequence similarity, which was commonly used for the last three decades, should be upgraded by phylogenetic analysis in genotyping of recombinant-free HBV strains. In this study, 4,429 HBV whole-genome sequences were employed to reconstruct the phylogeny of HBV using Bayesian inference. After excluding recombinant sequences, calculating partitioned evolutionary models, excluding recombinant sequences, reconstructing phylogenetic trees, and performing a correlation analysis of genetic distances, geographical distribution and serotypes, we systematically redefined the genotypes and subgenotypes of HBV. Compared to previous taxonomy, fourteen subgenotypes (A5-A7; B5-B9; C2-C4, C7; and D6-D7) were revised in the new standard. Now the HBV is divided into ten genotypes (A-J) and 24 subgenotypes (A1-A3; B1-B5; C1-C6; D1-D6; and F1-F4). Our robust genotype/subgenotype new taxonomy has objectively re-molded the current shape of HBV classification. We believe that all future hepatitis B related researches or diagnosis will be benefited under the new HBV genotyping/subgenotyping standards.


Conformational change in an engineered biliverdin-binding cyanobacteriochrome during the photoconversion process.
Cyanobacteriochromes (CBCRs) derived from cyanobacteria are linear-tetrapyrrole-binding photoreceptors related to the canonical red/far-red reversible phytochrome photoreceptors. CBCRs contain chromophore-binding cGMP-specific phosphodiesterase/adenylate cyclase/FhlA (GAF) domains that are highly diverse in their primary sequences and are categorized into many subfamilies. Among this repertoire, the biliverdin (BV)-binding CBCR GAF domains receive considerable attention for their in vivo optogenetic and bioimaging applications because BV is a mammalian intrinsic chromophore and can absorb far-red light that penetrates deep into the mammalian body. The typical BV-binding CBCR GAF domain exhibits reversible photoconversion between far-red-absorbing dark-adapted and orange-absorbing photoproduct states. Herein, we applied various biochemical and spectral studies to identify the details of the conformational change during this photoconversion process. No oligomeric state change was observed, whereas the surface charge would change with a modification of the α-helix structures during the photoconversion process. Combinatorial analysis using partial protease digestion and mass spectrometry identified the region where the conformational change occurred. These results provide clues for the future development of optogenetic tools.


Ovarian Suppression for Adjuvant Treatment of Hormone-Positive Early Breast Cancer.
Editor's note: The mission of Cochrane Nursing is to provide an international evidence base for nurses involved in delivering, leading, or researching nursing care. Cochrane Corner provides summaries of recent systematic reviews from the Cochrane Library. For more information, see https://nursing.cochrane.org.


[Comparative study of clinical and radiographic examinations for detection of proximal caries in posterior deciduous teeth].
Statistical analysis on data obtained through clinical and radiographical interproximal exams was the main purpose of this study. Posterior deciduous teeth of 450 children of both sex, of low social and economical level, without distinction of colour, aging seven to ten years were examined.


Are Europeans moving towards dietary habits more suitable for reducing cardiovascular disease risk?
Cardiovascular diseases are the main cause of death in Europe. Food choices represent the most important factors undermining health and well-being; they account for as much as half of all CVD deaths in Europe. The aim of this viewpoint is to evaluate food choices of the European population and their temporal trends in relation to possible effects on the cardiovascular disease risk. The CVD death rate attributable to diet-related factors has fallen in Europe over the last 25 years; however, the pace of the reduction has slowed down in the last ten years. In parallel, in the last decade unfavorable changes in the dietary habits associated with CVD risk have occurred. A mismatch exists between the available evidence on the health-promoting potential of the diet and the relatively modest and inconsistent improvements of dietary habits in the European population observed in recent years. Nutritional education alone will not be enough to improve the lifestyle of people in Europe. Policy options to be considered to reach this aim should necessarily include also initiatives for facilitating production, marketing, availability and affordability of healthy foods in each and every European country.


Presentation and Management of Uterine Didelphys with Unilateral Cervicovaginal Agenesis/Dysgenesis (CVAD): A Multicenter Case Series.
There are several well-described presentations of uterine didelphys (UD): UD without vaginal septum, UD with non-obstructed longitudinal vaginal septum, or UD with duplicated vaginas and an obstructed hemivagina on one side with ipsilateral renal anomaly. To describe another variant of UD and compare the presentation and management across different institutions METHODS: This was a retrospective case series approved by the NASPAG Fellows Research Consortium. Participating institutions obtained IRB approval. Inclusion criteria included a diagnosis of UD and unilateral cervicovaginal agenesis/dysgenesis (CVAD). Descriptive statistics were used. Five patients met the inclusion criteria, with ages ranging from 13 to 27 years. Presenting symptoms included dysmenorrhea (80%), irregular bleeding (40%), acute onset left lower quadrant pain (20%), and abdominal mass (20%). Three patients had additional known abnormalities, including solitary kidney and solitary adrenal gland. All patients underwent pelvic magnetic resonance imaging. Two cases were only suspicious for unilateral CVAD on imaging and required pathology review postoperatively to confirm diagnosis. Two cases required a 2-staged approach with an initial diagnostic surgery followed by a second definitive procedure. Three patients were noted to have endometriosis intraoperatively. Postoperative follow-up ranged from 2 months to 2 years, with 1 patient reporting chronic pelvic pain. Diagnosis on the basis of pelvic imaging can be difficult, as this unique variant may mimic classic obstructed hemivagina with ipsilateral renal anomaly. In patients with UD with unilateral CVAD, standard management is removal of the obstructed uterine horn. This multicenter series stresses awareness about the clinical presentation, distinguishes cases of cervical agenesis from dysgenesis, and reviews approaches to management.


Clinical characteristics of central serous chorioretinopathy in women.
To describe the clinical features of central serous chorioretinopathy (CSC) in women and identify factors predictive of complete recovery, moderate visual loss, and prolonged duration. Retrospective observational case series. Eighty-four eyes in 78 women with CSC seen in our practices between 1982 and 1999. Univariate and multivariate statistical analysis. Analyses of three outcome parameters: complete recovery (recovery of visual acuity and complete symptom resolution), moderate visual acuity loss (final acuity less than 20/40), and duration of symptoms greater than 5 months. Sixty-two percent (52 of 84) achieved complete recovery over a median of 5 months (range, 2-108 months) and 88% (74 of 84) had a final vision of 20/40 or better. Patients with subretinal precipitates (P = 0.001), single occurrence (P = 0.002), absence of hormone replacement therapy (HRT) (P = 0.01), duration less than 5 months (P = 0.02), or absence of a pigment epithelial detachment (PED) (P = 0.05) were more likely to recover completely. Recurrence (P = 0.03) and lack of subretinal precipitates (P = 0.03) were associated with a final vision less than 20/40. Age older than 50 (P = 0.004) and the presence of a PED (P = 0.02) were associated with duration longer than 5 months. In women, CSC associated with subretinal precipitates, shorter duration, single occurrence, lack of HRT use, and absence of PED is more likely to resolve completely. CSC occurring in women older than 50 or associated with PED formation is more likely to take longer to resolve.


The pharmacology of 14-hydroxyazidomorphine.
6-Deoxy-6-dihydroazido-14-hydroxyisomorphine (14-hydroxyazidomorphine) was synthesized, its analgesic potency in mice and rats, its antitussive effect in rats and its dependence liability in mice, rats and monkeys were studied. Azidomorphine the 14-nonhydroxylated parent molecule, morphine, hydromorphone and oxymorphone were used for comparison. 14-Hdroxyazidomorphine proved to be as potent an analgesic as azidomorphine, even more potent as an antitussive, and showed the same low tolerance and dependence capacity. It was 11-6 times less toxic than azidomorphine in mice and 6-5 times less toxic in rats.


Risk factors and the Child Behavior Checklist in a child mental health center setting.
The relationship between risk factors and the severity and type of childhood disorder, as measured by parent-completed Child Behavior Checklists, was examined for 768 children, ages 4-16, seen at a child mental health center. Regression analyses revealed no significant relationships between any combination of risk factors and the total number of behavior problems, internalizing, or externalizing scores for the entire group, males and females separately, or for the age and sex groupings of 6- to 11- and 12- to 16-year-olds. Chi-square tests revealed no relationship of profile type with any risk factor or with total number of risk factors. Implications for broad-based child assessment and risk factor research within a clinical population are discussed.


Neonatal disorders diagnosed with ultrasound.
This article discusses a large number of neonatal conditions for which ultrasound, by virtue of its noninvasiveness and its minimum of disturbance to the neonate, is the ideal mode for first investigation.


A hemoglobin-based oxygen carrier, bovine polymerized hemoglobin (HBOC-201) versus hetastarch (HEX) in a moderate severity hemorrhagic shock swine model with delayed evacuation.
To evaluate the efficacy of HBOC-201 for resuscitation of hemorrhagic shock in a swine model incorporating soft tissue injury and delayed evacuation. A muscle crush injury and 40% estimated blood volume controlled hemorrhage was completed in 24 Yucatan mini-pigs. Pigs were untreated or resuscitated with HBOC-201 or 6% hetastarch (HEX) at 20 min. Invasive hemodynamics and clinical variables were monitored for 4 h (pre-hospital phase) and subsequent fluid infusions were administered for severe hypotension or tachycardia. Animals were recovered from anesthesia and monitored non-invasively to 72 h (hospital phase). 100% (8/8) of HBOC-201-, 88% (7/8) of HEX-, and 63% (5/8) of non-resuscitated pigs, survived to 72 h (p=0.27). Mean arterial pressure, mean pulmonary arterial pressure and systemic vascular resistance index were higher in HBOC-201 pigs. By 90 min, cardiac index was restored to baseline in the HBOC-201 group and was 1.4-fold greater than baseline in the HEX group. HBOC-201 pigs had lower fluid requirements than HEX pigs (18.8+/-1.8 and 29.9+/-1.1 ml/kg, p<0.001) in the pre-hospital phase and required fewer blood transfusions (1.3+/-1.3 and 9.4+/-0.6 ml/kg, respectively, p<0.001) in the hospital phase. Urine output and blood creatinine were comparable in HBOC-201 and HEX pigs. Tissue oxygenation levels were highest in the HBOC-201 group. As HBOC-201 restored hemodynamics and tissue oxygenation and decreased fluid requirements, in comparison with HEX, HBOC-201 was at least as efficacious and possibly a superior resuscitative fluid in a military-relevant delayed evacuation hemorrhagic shock swine model.


Modulatory effects of sesamol in dinitrochlorobenzene-induced inflammatory bowel disorder in albino rats.
Inflammatory bowel disease (IBD) is a chronic inflammatory condition of gastrointestinal tract of immune, genetic and environmental origin. In the present study, we examined the effect of sesamol (SES), the main anti-oxidative constituent of Sesamum indicum (sesame seed) Linn. in the dinitrochlorobenzene (DNCB)-induced model for IBD in rats. The groups were divided into normal control, DNCB control, SES and sulfasalazine (SS). On day 24, the rats were killed, colon removed and the macroscopic, biochemical and histopathological evaluations were performed. The levels of MPO, TBARS and nitrite increased significantly (p < 0.05) in the DNCB group, whereas reduced significantly in the SES, SS treated groups. Serum nitrite levels were found to be insignificant between the different groups. IL-6 and TNF-α levels were significantly high in the DNCB group. We conclude the mucosal protective effect of SES on colon due to its potent antioxidant actions. Further investigation is required in a chronic model of different rodent strain for its role involved in the cytokine pathway.


Purification and detailed study of two clinically different human glucose 6-phosphate dehydrogenase variants, G6PD(Plymouth) and G6PD(Mahidol): Evidence for defective protein folding as the basis of disease.
In an attempt to investigate the molecular mechanism underlying human glucose-6-phosphate dehydrogenase (G6PD) deficiency caused by two mutations, G6PD(Plymouth) (G163D) and G6PD(Mahidol) (G163S), the two variants were constructed by site-directed mutagenesis and expressed in G6PD-deficient E. coli DF 213 cells. A first indication of impaired folding came from problems in expressing these clinical mutants, which were only overcome by lowering the growth temperature or co-expressing with molecular chaperones (GroEL and GroES). Both strategies significantly increased soluble expression of recombinant G6PD(Plymouth) and G6PD(Mahidol), judged by both G6PD activity in extracts and the amount of immunoreactive protein. Using a modified 3-step protocol, the two mutant enzymes were successfully purified for the first time. Steady-state kinetic parameters (K(m) for NADP(+), K(m) for G6P and k(cat)) of the two mutants are very similar to the wild-type values, indicating that the catalytic efficiency of the two mutants remains unchanged. The two mutants are, however, markedly less stable than wild-type G6PD in both thermostability and urea-induced inactivation tests. In a typical experiment at 37 degrees C and pH 7.2 after 24h G6PD WT, G6PD(Mahidol) and G6PD(Plymouth) retained 58.3%, 27.0% and 3.9%, respectively, of their corresponding initial activity. The stability of all three enzymes is enhanced by addition of NADP(+). According to unfolding and refolding experiments, the two mutants are impaired in their folding properties. Thus structural instability appears to be the molecular basis of the clinical phenotype in G6PD(Plymouth) and G6PD(Mahidol) and in particular of the differing clinical severity of the two mutations. The 3-D structure solved for G6PD(Canton) allows an interpretation of these effects in terms of steric hindrance.


The unfolding story of a redox chaperone.
Oxidative stress, especially in combination with heat stress, poses a life-threatening challenge to many organisms by causing protein misfolding and aggregation. In this issue, Reichmann et al. demonstrate how a destabilized linker region of the bacterial chaperone Hsp33 prevents aggregation of a denatured protein by stabilizing structural elements.


The recovery of mammalian cells treated with methyl methanesolfonate, nitrogen mustard or UV light. II. The importance of DNA repair prior to the initiation of S phase.
CHO cells were synchronized 2 G1 phase and treated with UV light or HN2. These treatments resulted in a dose-dependent reduction in the rate of DNA replication and cell survival. Holding UV-irradiated cells in G1 phase (in HU medium) for an additional 10 h prior to their release into S phase did not assist recovery as measured by either of these criteria. The survival of cells treated with HN2 was also not enhanced by this recovery period. However, following 2 X 10(-5) M HN2 the rate of DNA replication increased from 30% to 70% of the control level when the period in HU medium was extended to 14 h. The induction of cross-links following HN2 treatment of asynchronous cells was shown to be dose dependent. Subsequent incubation in fresh medium resulted in complete recovery within 20 h at concentrations of HN2 up to 10(-5) M, and at 2 X 10(-5) M HN2, 75% of the cross-links were removed at 14 h post treatment.


Dietary Risk Factors and Odds of Colorectal Adenoma in Malaysia: A Case Control Study.
Current evidence suggests that dietary and lifestyle factors may play an important role in colorectal cancer risk but there are only a few studies that investigated their relationship with colorectal adenomas (CRA), the precursors for colorectal cancer. A case-control study was conducted to determine the relationship between dietary and lifestyle factors associated with CRA risk among 125 subjects with CRA and 150 subjects without CRA at Hospital Canselor Tuanku Muhriz UKM (HCTM), Malaysia. We used dietary history questionnaire (DHQ) and International Physical Activity Questionnaire-Short Form (IPAQ) to estimate the diet and physical activity. The findings of this study showed that male gender [OR = 2.71 (95% CI= 1.01-7.27)], smoking [OR = 6.39 (95% CI= 1.04-39.30)], family history of cancer [OR = 6.39 (95% CI= 1.04-39.30)], high body fat percentage [OR = 1.25 (95% CI= 1.04-1.51)], high calorie and fat intake [OR = 1.03 (95% CI= 1.01-1.06)], [OR = 1.01 (95% CI= 0.95-1.09)] and red meat intake more than 100 g per day [OR = 1.02 (95% CI= 1.01-1.04)] increased CRA risk. High fiber [OR = 0.78 (95% CI= 0.64-0.95)] and calcium intake [OR = 0.78 (95% CI= 0.98-1.00)] was found to decrease CRA risk. Some of these modifiable risk factors could be advocated as lifestyle interventions to reduce risk of CRA.


Patterned polymer matrix promotes stemness and cell-cell interaction of adult stem cells.
The interaction of stem cells with their culture substrates is critical in controlling their fate and function. Declining stemness of adult-derived human mesenchymal stem cells (hMSCs) during in vitro expansion on tissue culture polystyrene (TCPS) severely limits their therapeutic efficacy prior to cell transplantation into damaged tissues. Thus, various formats of natural and synthetic materials have been manipulated in attempts to reproduce in vivo matrix environments in which hMSCs reside. We developed a series of patterned polymer matrices for cell culture by hot-pressing poly(ε-caprolactone) (PCL) films in femtosecond laser-ablated nanopore molds, forming nanofibers on flat PCL substrates. hMSCs cultured on these PCL fiber matrices significantly increased expression of critical self-renewal factors, Nanog and OCT4A, as well as markers of cell-cell interaction PECAM and ITGA2. The results suggest the patterned polymer fiber matrix is a promising model to maintain the stemness of adult hMSCs. This approach meets the need for scalable, highly repeatable, and tuneable models that mimic extracellular matrix features that signal for maintenance of hMSC stemness.


Comment on "Topically Applied Connective Tissue Growth Factor/CCN2 Improves Diabetic Preclinical Cutaneous Wound Healing: Potential Role for CTGF in Human Diabetic Foot Ulcer Healing".
A recent paper in this journal, presented a novel method by topical application of growth factors in stimulating diabetic cutaneous wound healing that caught our attention. We believe that the experimental method in the article is efficient and creative, but it also has some controversies and shortcomings to be discussed. We noted that the authors used "Tegaderm" as a semiocclusive dressing film and stated that it exerted a "splinting effect" on the wound margins and controlled contraction. Indeed, the "Tegaderm" itself can serve as a dressing film to isolate the wound bed with outside environments while the "splinting effect" is mainly achieved by adding silicone splints around the wound. Considering the unique properties of silicone splints and "Tegaderm," our experimental group propose an alternative method named "combined-suturing" technique that is not only suturing the silicone splints but also securing the "Tegaderm" around the wound. The specific reasons and operative procedures are explained in detail in this letter.


Investigating changes in travel behavior over time in response to the COVID-19 pandemic.
The COVID-19 pandemic has significantly affected travel behavior, including the frequency and mode of travel, with the magnitude and nature of these effects varying over time. This study investigates the nature of these relationships by examining changes in various measures of travel behavior, including weekly driving hours, as well as the frequency of telecommuting, use of ride-sharing services, travel for medical purposes, and use of food delivery services. Self-reported travel data from a representative statewide survey of Michigan residents were used to assess changes in these metrics during the early stages of the pandemic, as well as one year thereafter. Random effects linear regression and ordered logit regression models were estimated and the findings show that various changes in behavior had long-lasting effects, while other behaviors generally reverted back toward pre-pandemic levels. In addition, these changes were found to vary across individuals. For example, significant differences were observed based on socio-demographic characteristics, between urban and rural areas, and amongst individuals with differing views on COVID-19 and related government interventions. In general, the pandemic tended to have less pronounced and sustained effects among younger adults as compared to older age groups. Further, those individuals who were opposed to mandatory COVID-19 vaccines were less likely to change their travel behavior, during both the early and latter stages of the pandemic. Changes were observed consistently across most of the travel metrics of interest. Among these, overall driving hours, travel for medical purposes, and ride-sharing were still lower during the latter stages of the pandemic, while telecommuting and the use of food delivery services reverted nearer to pre-pandemic levels.


The effect of montelukast on wheal reactions in skin prick tests: a double-blind-placebo-controlled randomized trial.
It is well-known that number of drugs may interfere with wheal reactions in skin prick test. However, the effect of long-term use of montelukast, a cystenil leukotriene receptor antagonist, on skin prick test hasn't been full elucidated. The aim of present study was to demonstrate the effect of montelukast on skin prick tests (SPT). This is a single-center, randomized, double-blinded, placebo-controlled study including two treatment periods with a wash-out interval. The subjects received montelukast (5 mg per day), fexofenadine HCl (60 mg per day) and placebo (lactose) with a double-blinded manner during 7- and 21-days treatment periods with a 14 days wash-out period. Dermatophagoides farinae (D. farinae) was used as the skin test material, while histamine as positive control and normal saline as negative control. Overall, 7 skin prick tests were performed at following time points: before treatment periods, on the last days of both treatment periods, 24 h after completion of treatment periods, and on the last day of 14-days interval. Sixty house dust mite (HDM) allergic children (23 girls and 37 boys) with allergic rhinitis and/or asthma completed the study. Mean age was 8.3 ± 2.0 years. In the fexofenadine group, a significant suppression was observed in post-treatment values when compared to baseline values in SPT with D. farinae (p = 0.019). In the montelukast group, no significant suppression was observed in SPT with D. farinae at all time points when compared to baseline. Our results showed that montelukast had no effect on measurements of SPT. Thus, we concluded that there is no need to discontinue the treatment in order to perform SPT in patients receiving montelukast, even in those on montelukast treatment for at least 21 days.


A Novel Chd8 Mutant Mouse Displays Altered Ultrasonic Vocalizations and Enhanced Motor Coordination.
Mutations in CHD8 are among the most common autism-causing genetic defects identified in human genomics studies. Therefore, many labs have attempted to model this disorder by generating mice with mutations in Chd8. Using a gene trap inserted after Exon 31, we created a novel Chd8 mutant mouse (Chd8+/E31T ) and characterized its behavior on several different assays thought to have face validity for the human condition, attempting to model both the core symptoms (repetitive behaviors and social communication impairments) and common comorbidities (motor deficits, anxiety, and intellectual disability). We found that Chd8+/E31T mice showed no difference compared to wild-type mice in amount of self-grooming, reproducing the negative finding most other studies have reported. Unlike some of the other published lines, Chd8+/E31T mice did not show deficits in the three-chamber test for social novelty preference. A few studies have examined ultrasonic vocalizations in Chd8 mutant mice, but we are the first to report an increase in call length for adult mice. Additionally, we found that in contrast to previous published lines, Chd8+/E31T mice displayed no anxiety-like behaviors or learning impairments but showed paradoxically significant improvement in motor function. The inconsistencies in behavioral phenotypes in the Chd8 mutant mice generated by different laboratories poses a challenge for modeling autism spectrum disorder and preclinical studies in mice going forward and warrants further investigation into the molecular consequences of the different mutations in Chd8 and the functional impact on behavior. LAY SUMMARY: Several different mouse models carrying mutations in the Chd8 gene have been created to study the effects of these autism-causing mutations in the laboratory. The current study characterizes a novel Chd8 mutant mouse model as well as summarizes data from previously published Chd8 mutant mice. The inconsistencies between different studies are concerning, but future research into the reasons why these inconsistencies occur may help us understand why patients with various mutations have different degrees of symptom severity. Autism Res 2020, 13: 1685-1697. © 2020 International Society for Autism Research and Wiley Periodicals LLC.


Meniscal Injuries Are Decreasing but Are Increasingly Being Treated Surgically With Excellent Return to Play Rates in Professional Baseball Players.
The purposes of this study were to determine the incidence and key characteristics of meniscus injuries in professional baseball players, assess current treatment strategies, determine the return to play rates at any level (RTP) and at the same level (RSL), and identify prognostic factors that predict injury severity. After approval from the Major League Baseball (MLB) Research Committee and our institutional review board, the MLB Health and Injury Tracking System was used to identify meniscus injuries occurring across MLB and Minor League Baseball (MiLB) from 2011 to 2017. Analyzed injuries occurred during normal baseball activity in a player who was active on an MLB or MiLB roster and resulted in at least 1 day missed. A total of 293 professional baseball players sustained 314 meniscus injuries from 2011 to 2017 (7 years) for a mean of 44.9 injuries/y. Pitchers were the most injured position (31.8%), followed by infielders (26.4%). Catchers and infielders missed the most median number of days (50 days). When comparing injuries to landing leg vs push-off leg in pitchers, injury to the push-off leg resulted in significantly more days missed per injury compared to the lead leg (59.6 vs 39.9 days, P = .048). Overall, RTP was 93.0%, while RSL was 84.4%. Over 7 professional baseball seasons, 314 meniscus injuries occurred in 293 players. Pitchers and catchers were most injured, and overall, the number of meniscal injuries per year declined while the percentage of injuries that required surgery increased over time. High rates of RTP were observed. Level IV, therapeutic case series.


Production of aflatoxin and cyclopiazonic acid by various aspergilli: An ELISA analysis.
An enzyme-linked Immunosorbent assay (ELISA) was used to monitor a total of 153 fungi in theAspergillus flavus group, Including 130A. flavus, 15A. parasiticus and 8A. tamarii, for their ability to produce aflatoxins (AFs) and cyclopiazonic acid (CPA) in a mycologlcal broth-sucrose-yeast extract medium. Of 15A. parasiticus isolates, ten produced AFs In a range of 12.4 to 89.3 μg/vial (average 56.9 μg/vial); two isolates produced only trace amounts of AFs and three isolates produced none at all. Production of CPA was not demonstrated in anyA. parasiticus isolate. On the other hand, all A. tamarii isolates produced only CPA with a range of 310 to 1100 gmg/vial. Fifteen percent (14.6%) of theA. flavus isolates (19/130) produced more than 500 μg CPA/vial, but yielded no or little AF (less than 0.1 μg/vial). About 22.3% ofA. flavus (29/130) that produced less than 500 μg of CPA also yielded little or no aflatoxin. MostA. flavus isolates (44.6%) produced both CPA (50 to 300 μg/vial) and AFs (10 to 40 μg/vial). About 9.2% of theA. flavus are low CPA producers (less than 100 μg/vial) but yielded higher amounts of AFs. A small percentage (12/130 or 9.2%) of A. flavus isolates produced neither CPA nor aflatoxin. Excluding the isolates that produced neither AFs nor CPA, there is a negative correlation between the production of CPA and AFs by most A.flavus isolates. Data obtained from ELISA for the production of CPA were consistent with TLC results. Thus, the ELISA method for CPA and AFB could be applied to the screening of toxigenic fungi. Data on the simultaneous production of both toxins by a large percentage of the toxigenicA. flavus isolates suggest that there is a potential health hazard for co-existence of both toxins in foods and feeds.


Hollow fibers as an oral sustained-release delivery system.
Phenylpropanolamine (PPA) bound to ion-exchange resin was encapsulated in hollow fibers made of segmented polyurethane. This system was examined as an oral sustained-release delivery system. The fibers were spun by the phase inversion process and cut into different aspect ratios (length/diameter). The U.S.P. basket dissolution method was used to evaluate the in vitro drug release kinetics and the effect of the aspect ratio on the release. For in vivo evaluation, selected fibers were orally administered to dogs in gelatin capsules. The fiber delivery system provided a sustained-release profile of plasma PPA and a longer terminal half-life when compared to an oral immediate-release formulation.


Evidence against the occurrence of artifacts due to carrier ampholyte-protein binding during isoelectric focusing.
The formation of irreversible complexes between carrier ampholyte components and proteins was investigated by gel filtration of mixtures of proteins and radioactively labelled ampholytes. Experiments were performed both with purified proteins (albumin, ferritin, beta-glucuronidase) and with a complex mixture of proteins (serum); in no case was binding of ampholytes to proteins detected. Thus the results argue against the occurrence in isoelectric focusing of proteins of artifacts due to such complex formation.


Use of the collagen-binding assay for von Willebrand factor in the analysis of type 2M von Willebrand disease: a comparison with the ristocetin cofactor assay.
This study compares the utility of two functional assays for von Willebrand factor (VWF), the ristocetin cofactor assay (VWF:RCo) and the collagen-binding assay (VWF:CBA). We analysed a group of 32 patients with type 2 von Willebrand disease (VWD) (25 patients with type 2M, six with type 2A and one with type 2B) and 22 normal control subjects. VWF:RCo/VWF antigen (VWF:Ag) ratios and VWF:CBA/VWF:Ag ratios were compared between the patient and control groups. In the six patients with type 2A VWD, both VWF:RCo/VWF:Ag ratios and VWF:CBA/VWF:Ag ratios were discordant (< or = 0.7). In the 25 type 2M VWD patients, the VWF:CBA/VWF:Ag ratios were concordant (> 0.7), but the VWF:RCo/VWF:CBA ratios were discordant (< or = 0.7) (P = 0.001) compared with control subjects. Thus, VWF:RCo/VWF:Ag ratios were discordant in both type 2M and 2A VWD patient groups indicating a functional abnormality. However, VWF:CBA/VWF:Ag ratios were discordant in the type 2A VWD group but not in the type 2M VWD group. Our study showed that VWF:CBA is sensitive to functional variants associated with the loss of high-molecular-weight multimers, i.e. type 2A and 2B in VWD, but the assay was unable to discriminate defective platelet-binding VWD variants with normal multimeric patterns such as type 2M VWD. It was concluded that the VWF:CBA assay should be used in association with rather than as a replacement for the VWF:RCo assay.


A case of restitution ad integrum in Gorham-Stout disease.
Gorham-Stout disease (or "vanishing bone" disease) is a rare mono or polyostotic disease of unknown etiology, characterized by intraosseous angiomatous proliferation leading to bone resorption. We report the case of a 17-year-old woman presenting with symptomatic osteolytic lesions of the frontal vault. Imaging was suggestive of Gorham-Stout disease without argument for other diagnoses. An unusual evolution of the "vanishing bone" lesions was observed on the scan after one year, with a full recovery of the lytic lesions. This report shows for the first time a spontaneous restitutio ad integrum of bone matrix in Gorham-Stout disease.


Possible beta-adrenoceptor modulating effect of ZK 36 374 a stable analogue of carbacyclin.
Experiments were performed to investigate the nature of the interaction of noradrenaline (NA) and adrenaline (A) with ZK 36 374, a stable analogue of carbacyclin, in spontaneously beating isolated rat right atria and spirally cut rabbit aortic strips. At lower concentrations (below 10(-8) M), ZK 36 374 failed to alter heart rate and contractility but caused an inhibition of both NA effects. At higher concentrations (above 10(-7) M), the compound produced a weak positive chronotropic and inotropic response but did not cause a further decrease in the responses to NA. At the same concentrations propranolol (PR) produced more pronounced inhibition of NA actions. The nature of the inhibition by ZK 36 374 against NA in the rat atria seems to be different from that of PR since the calculated pD2 values of NA were the same for chronotropic and inotropic responses in control experiments and in the presence of PR while the pD2 value of NA for chronotropic response was significantly lower than that for inotropic response in the presence of ZK 36 374. Nicotine partly reversed the inhibitory effect of PR against NA on heart rate which was prevented by ZK 36 374. ZK 36 374 caused a significant potentiation of the contractile response to A but not to NA in rabbit aortic strips. These results suggest that ZK 36 374 interacts with beta-adrenoceptors causing an inhibition on the responses of beta-mediated pharmacological effects of A and NA. These results were also taken as an evidence that PR may partly act through the release of endogenous PGI2.


Modified sandwich embolization technique for postpartum hemorrhage caused by uterine artery pseudoaneurysm: a case series.
Uterine artery pseudoaneurysm (UAP) is rare but can cause life-threatening postpartum hemorrhage (PPH). To evaluate a novel sandwich embolization technique as a treatment for PPH caused by UAP. This retrospective study included 10 patients with PPH caused by UAP who were treated using a modified sandwich embolization technique at the Radiology Department, Third Affiliated Hospital of Guangzhou Medical University between April 2009 and September 2018. Baseline clinical characteristics, intraoperative data (including treatment effectiveness) and postoperative data (including re-bleeding events and complications) were extracted from the medical records. Uterine arterial angiography showed cystic shadowing of the vascular wall during the arterial phase in all patients. Spraying of contrast agent into the pseudoaneurysm was observed for large UAPs, and the pseudoaneurysm disappeared in the venous phase. The pseudoaneurysm blood supply was from the uterine artery in 9 patients (90%) and the uterine, superior vesical, internal pudendal and nameless little arteries in 1 patient (10%). Bleeding symptoms were completely relieved in all patients after sandwich embolization. Eight patients experienced painful contractions in the perioperative period, but there were no other postoperative complications. During the 1-year postoperative follow-up, 9 patients (90%) had no re-bleeding symptoms/signs. One patient (10%), who had a pseudoaneurysm supplied by the uterine, superior vesical, internal pudendal and nameless little arteries, experienced re-bleeding 20 days after surgery and was treated by hysterectomy. Modified sandwich embolization is an effective treatment for PPH caused by UAP.