DGGE with genomic DNA: suitable for detection of numerically important organisms but not for identification of the most abundant organisms.
Identification of all important community members as well as of the numerically dominant members of a community are key aspects of microbial community analysis of bioreactor samples. A systematic study was conducted with artificial consortia to test whether denaturing gradient gel electrophoresis (DGGE) is a reliable technique to obtain such community data under conditions where results would not be affected by differences in DNA extraction efficiency from cells. A total of 27 consortia were established by mixing DNA extracted from Escherichia coli K12, Burkholderia cepacia and Stenotrophomonas maltophilia in different proportions. Concentrations of DNA of single organisms in the consortia were either 0.04, 0.4 or 4ng/microl. DGGE-PCR of genomic DNA with primer sets targeted at the V3 and V6-V8 regions of the 16S rDNA failed to detect the three community members in only 7% of consortia, but provided incorrect information about dominance or co-dominance for 85% and 89% of consortia with the primer sets for the V6-V8 and V3 regions, respectively. The high failure rate in detection of dominant B. cepacia with the primers for the V6-V8 region was attributable to a single nucleotide primer mismatch in the target sequences of both, the forward and reverse primer. Amplification bias in PCR of E. coli and S. maltophilia for the V6-V8 region and for all three organisms for the V3 region occurred due to interference of genomic DNA in PCR-DGGE, since a nested PCR approach, where PCR-DGGE was started from mixtures of 16S rRNA genes of the organisms, provided correct information about the relative abundance of original DNA in the sample. Multiple bands were not observed in pure culture amplicons produced with the V6-V8 primer pair, but pure culture V3 DGGE profiles of E. coli, S. maltophilia and B. cepacia contained 5, 3 and 3 bands, respectively. These results demonstrate DGGE was suitable for identification of all important community members in the three-membered artificial consortium, but not for identification of the dominant organisms in this small community. Multiple DGGE bands obtained for single organisms with the V3 primer pair could greatly confound interpretation of DGGE profiles.


[Duplex ultrasound of the liver and portal vein system].
Duplex ultrasound allows direct access to portal venous and hepatic vein hemodynamics. This method has been accepted as a reliable procedure to detect portal and splenic venous thrombosis, Budd-Chiari's syndrome, portosystemic collateralisation and changes in the portal venous blood flow direction with a high rate of sensitivity. Besides these qualitative findings Doppler flowmetry allows quantitative measurement of portal venous blood flow velocities. The estimation of portal hypertension and the prediction of the risk of variceal bleeding by quantitative Doppler flowmetry is of increasing clinical importance. In monitoring the efficiency of medicational prophylaxis against gastrointestinal bleeding is also helpful. Duplex ultrasound can be used to evaluate the function of portocaval shunts including the intrahepatic stent shunt as well as to detect vascular complications after liver transplantation. Furthermore the method is gaining importance in the diagnosis and monitoring of therapy of acute liver transplant rejection. In case of liver tumors duplex ultrasound is of value for tumor staging and provides information about the dignity on the basis of perfusion patterns.


Lyophilized Cheliensisin A submicron emulsion for intravenous injection: characterization, in vitro and in vivo antitumor effect.
Growing attentions have been focused on natural antitumor drugs. Recently, a novel and potent antitumor drug Cheliensisin A (GC-51) with broad-spectrum efficiency has been developed. However, due to its poor water solubility and chemical instability, choosing the appropriate dosage form is of great significance. This study aimed at developing a lyophilized submicron emulsion for GC-51 and further improving the therapeutic index of the drug. The resultant lyophilized GC-51 submicron emulsion was much more stable than its solution, which can be stored for years without significant change on physicochemical properties. And its solubility was increased from 6.74+/-0.14 to 2.00+/-0.10 mg mL(-1). The 50% inhibitory concentration IC50 values were calculated from growth curves by MTT assay on various tumor cell lines. Compared with the IC50 of GC-51 crude drug, that of lyophilized GC-51 submicron emulsion decreased from 24.04+/-1.97 to 8.23+/-1.84 microg mL(-1) on HepG2, and from 31.08+/-2.56 to 10.85+/-2.09 microg mL(-1) on CT-26, from 17.90+/-1.83 to 7.49+/-1.87 microg mL(-1) on HeLa and from 16.38+/-2.41 to 10.13+/-2.12 microg mL(-1) on A549, respectively. In the time-dependent assay of tumor cell viability, lyophilized GC-51 submicron emulsion exhibited significantly lower inhibition rate in the initial action times, but increased gradually afterwards. That means lyophilized submicron emulsion as the vector for GC-51 had some protective and delayed release effect. Further, the in vivo therapeutic efficacy was measured in pulmonary metastasis of colon cancer-bearing BALB/c mice model. An obvious enhanced antitumor activity was observed after administration of lyophilized GC-51 submicron emulsion (P<0.05), which increased from 22.78+/-3.5 to 41.42+/-4.2% compared with GC-51 injection. And the life span of tumor-bearing mice in lyophilized GC-51 submicron emulsion group was significantly longer than that of the mice in GC-51 injection and normal saline groups. Compared with crude drug, the lyophilized GC-51 submicron emulsion showed a significantly higher antitumor efficiency both in vivo and in vitro, suggesting a potential application in tumor chemotherapy.


An adenovirus vectored mucosal adjuvant augments protection of mice immunized intranasally with an adenovirus-vectored foot-and-mouth disease virus subunit vaccine.
Foot-and-mouth disease virus (FMDV) is a highly contagious pathogen that causes severe morbidity and economic losses to the livestock industry in many countries. The oral and respiratory mucosae are the main ports of entry of FMDV, so the stimulation of local immunity in these tissues may help prevent initial infection and viral spread. E. coli heat-labile enterotoxin (LT) has been described as one of the few molecules that have adjuvant activity at mucosal surfaces. The objective of this study was to evaluate the efficacy of replication-defective adenovirus 5 (Ad5) vectors encoding either of two LT-based mucosal adjuvants, LTB or LTR72. These vectored adjuvants were delivered intranasally to mice concurrent with an Ad5-FMDV vaccine (Ad5-A24) to assess their ability to augment mucosal and systemic humoral immune responses to Ad5-A24 and protection against FMDV. Mice receiving Ad5-A24 plus Ad5-LTR72 had higher levels of mucosal and systemic neutralizing antibodies than those receiving Ad5-A24 alone or Ad5-A24 plus Ad5-LTB. The vaccine plus Ad5-LTR72 group also demonstrated 100% survival after intradermal challenge with a lethal dose of homologous FMDV serotype A24. These results suggest that Ad5-LTR72 could be used as an important tool to enhance mucosal and systemic immunity against FMDV and potentially other pathogens with a common route of entry.


A state-of-art review on camel milk proteins as an emerging source of bioactive peptides with diverse nutraceutical properties.
The generation of camel milk derived bioactive peptides (CM-BAPs) have started to grab keen interest of many researchers during the past decade. CM-BAPs have shown more significant bioactive properties in comparison to camel milk intact proteins. CM-BAPs can be obtained using enzyme hydrolysis to form hydrolysates, or by the fermentation process. In this systematic review, 46 research articles exploring the health-related bioactive properties of CM-BAPs through in-vitro and in-vivo studies have been included. CM-BAPs have been reported for their antioxidant, anti-diabetic, anti-obesity, antihypertensive, antibacterial, antibiofilm, anticancer, anti-inflammatory, anti-haemolytic, and anti-hyperpigmentation activities. The effects of factors such as molecular weight of peptides, type of enzyme, enzyme to substrate ratio, hydrolysis temperature and duration have been analysed. The in-vitro studies have provided enough evidence on certain aspects of the pharmacological actives of camel milk bioactive peptides. Nevertheless, the in-vivo studies are very limited, and no clinical studies on CM-BAPs have been reported.


Molecular diagnosis of ochratoxinogenicAspergillus species.
Toxigenic and non-toxigenic black aspergilli belonging to theAspergillus niger aggregate and toA. carbonarius were compared to each other and to strains of other species by DNA fingerprinting. AFLPs showed a clear separation ofA. niger andA. carbonarius. However, no clear correlation between the genetic similarity of the strains and the ability to produce ochratoxin A (OTA) was detected. Based on AFLP, marker sequences were chosen for the construction of SCAR-PCR primers for the detection ofA. carbonarius. A similar approach was used forA. ochraceus, another fungus of concern regarding ochratoxin A contamination of coffee. Cluster analysis ofA. ochraceus isolates mainly from Brazilian coffee showed a very close genetic similarity. Three species specific primer pairs were developed and one of these was used for the PCR and realtime PCR (RT-PCR) based detection of the mould in green coffee.A. ochraceus was specifically and rapidly detected and quantified in green coffee. A positive correlation between the amount of DNA and OTA content was established.


Serum p24 antigen level as an intermediate end point in clinical trials of zidovudine in people infected with human immunodeficiency virus type 1. Aids Clinical Trials Group Virology Laboratories.
Serum p24 antigen levels were examined in subjects from three clinical trials of zidovudine to determine whether the pattern of change in serum p24 antigen during the first 8-16 weeks of therapy was associated with human immunodeficiency virus type 1 (HIV-1) disease progression or death. Among 406 patients with AIDS and a first episode of Pneumocystis carinii pneumonia, 65% had measurable pretreatment concentrations of serum p24 antigen (> or = 10 pg/mL). Changes during treatment were not associated with reduced mortality. In 637 mildly symptomatic patients, 24% had measurable concentrations, and changes were marginally associated with increased time until more advanced disease. Among 683 asymptomatic patients, 18% had measurable concentrations, and changes were not associated with increased time until progression. Despite the small number of clinical events and the low rate of serum p24 antigen positivity in the latter two studies, pretreatment serum p24 antigen levels were predictive of clinical outcome; subsequent measurements appear to be of limited use in evaluating zidovudine therapy.


Impact of traditional Samoan lifestyle (fa'aSamoa) on cancer screening practices.
The Samoan way of life, or the fa'aSamoa, is a collection of practices and rituals by which Samoans conduct their lives. The purpose of this study was to explore the possible influence of the fa'aSamoa on cancer prevention practices. The investigators conducted six focus groups; three in Carson, California and three in Pago Pago, American Samoa among 60 men from the communities, matai (chiefs), faifeau (pastors). Using a discussion guide, bilingual Samoan moderators asked questions about the fa'aSamoa, how traditional cultural practices might influence cancer prevention, and whether certain aspects of the fa'aSamoa could be incorporated into cancer screening programs to improve screening rates. Through qualitative content analysis, the investigators identified major themes and came to a consensus about them. The mean age of the 60 participants was 61 years, all were married, 50 were born in American Samoa, and 10 were born in the Independent State of Samoa. Among the themes, three were particularly important. First, participants confirmed that the fa'aSamoa remains an important concept in Samoan life. Second, they believed that disease prevention was not an integral part of the fa'aSamoa. Indeed, traditionally modesty and reluctance to discuss personal issues may inhibit use of screening services. Despite these a beliefs, a third theme emerged. Because of the influence of the matai and faifeau in the fa'aSamoa, participants stressed that seeking input from these leaders could improve cancer prevention programs. The lack of emphasis on disease prevention in the fa'aSamoa may contribute to low cancer screening rates. However, other aspects of the culture, such as the respect for matai and faifeau could be incorporated into cancer prevention programs aimed at improving cancer control among Samoans.


Aptamer-Drug Conjugates of Active Metabolites of Nucleoside Analogs and Cytotoxic Agents Inhibit Pancreatic Tumor Cell Growth.
Aptamer-drug conjugates (ApDCs) have the potential to improve the therapeutic index of traditional chemotherapeutic agents due to their ability to deliver cytotoxic drugs specifically to cancer cells while sparing normal cells. This study reports on the conjugation of cytotoxic drugs to an aptamer previously described by our group, the pancreatic cancer RNA aptamer P19. To this end, P19 was incorporated with gemcitabine and 5-fluorouracil (5-FU), or conjugated to monomethyl auristatin E (MMAE) and derivative of maytansine 1 (DM1). The ApDCs P19-dFdCMP and P19-5FdUMP were shown to induce the phosphorylation of histone H2AX on Ser139 (γ-H2AX) and significantly inhibited cell proliferation by 51%-53% in PANC-1 and by 54%-34% in the gemcitabine-resistant pancreatic cancer cell line AsPC-1 (p ≤ 0.0001). P19-MMAE and P19-DM1 caused mitotic G2/M phase arrest and inhibited cell proliferation by up to 56% in a dose-dependent manner when compared to the control group (p ≤ 0.001). In addition, the cytotoxicity of P19-MMAE and P19-DM1 in normal cells and the control human breast cancer cell line MCF7 was minimal. These results suggest that this approach may be useful in decreasing cytotoxic side effects in non-tumoral tissue.


Do social capital and the quality of institutions affect waste recycling?
In a social capital framework, the quality of institutions and generalised social trust enhance attitudes towards disposal practices that are less environmentally invasive for the. This study aims to improve our empirical knowledge of this relationship through panel data models to show that waste recycling is affected by social capital and government effectiveness. The study uses Italian annual provincial data (NUTS-3 level) for 2004-2016 period to show that social capital and the quality of institutions are extremely important in determining waste recycling behavior. We find that both social capital and the quality of institutions, which act as 'substitutes for goods', increase waste recycling, although with different effects at the territorial level. In the southern and central areas, waste recycling needs to be spurred through policy interventions aimed at improving the quality of institutions, education and trust among citizens, whereas in northern Italy, social capital on its own is sufficient to enhance waste recycling. Hence, our results suggest that policies for waste recycling cannot be uncoupled from the promotion of social cohesion and institutional quality.


Prosthetic mesh "slim-cigarette like" for laparoscopic repair of ventral hernias: a new technique without transabdominal fixation sutures.
Prosthetic mesh rolled up and fixed with stitches like a slim cigarette ("slim-mesh") for laparoscopic ventral hernia (VH) repair is an new technique which allows an easy intraperitoneally introduction, distension and circumferential fixation of a prosthetic mesh without transabdominal fixation sutures even for meshes larger than 16 cm up to 30 cm for the "slim-mesh" repair of wide ventral hernias. We report the technique of laparoscopic repair of VH with "slim-mesh". This technique enables an easy intra-peritoneally introduction of the mesh through the trocar because it reduces consistently its size, it allows a rapid intra-abdominal handling of the mesh and a fast and easy fixation for VH repair. The average time of surgery with "slim-mesh" for treatment of all 28 VH was 97 min ranging from 57 to 160 min. The average time for the repair of the 24 VH smaller than 10 cm was 91 and 135 min for the four VH larger than 10-22 cm. This new surgical technique leads to a reduction of surgical risks avoiding the use of transfascial sutures with the associated complications. This new surgical procedure in our experience is fast, safe, simple and also easily reproducible by surgeons in laparoscopic training. This technique may be used in wide VH (larger than 10-22 cm) that generally require open surgery.


Analysis of dimeric species derived from the reaction of phosphatidylethanolamine with dimethylsuberimidate.
Phosphatidylethanolamines of various fatty acyl species in vortexed lipid dispersions were reacted with dimethylsuberimidate to produce dimeric products. The yield was 34% at pH 10 and 2% at pH 7. The crosslinked phosphatidylethanolamine species were separated from minor products and the reactants by extraction and two-dimensional thin-layer chromatography on silica gel G, gel filtration on lipophilic Sephadex, or C18-reversed phase high-performance liquid chromatography (HPLC). Reversed phase HPLC was also used to resolve the dimers into individual molecular species. Analysis of the dimers revealed the extinction coefficients at 205 nm to be higher than those of the reactant phosphatidylethanolamines. Proton nuclear magnetic resonance spectroscopy and chemical analysis implied that the dimers contain an amidine group.


Triacylglycerol hydroperoxides not detected in pig small intestinal epithelial cells after a diet rich in oxidized triacylglycerols.
The presence of TAG hydroperoxides in the epithelial cells of the small intestines in growing pigs was studied after they had consumed a diet rich in either nonoxidized or oxidized sunflower seed oil (PV in oils, 1 and 190 mequiv O2/kg, respectively). To obtain molecular-level information on the oxidized TAG structures, a new approach based on TLC and HPLC-electrospray ionization-MS was used in the analysis of the samples. TAG hydroperoxides were not detected in the small intestinal mucosa or adipose tissue of either group, whereas TAG hydroxides, ketones, and epoxides were detected in all samples. The results suggest that dietary TAG hydroperoxides do not lead to the appearance of these molecules in the tissues.


The Sesame-Peanut Conundrum in Israel: Reevaluation of Food Allergy Prevalence in Young Children.
Food allergies (FAs) are on the rise worldwide. A previous cross-sectional study from 2002 in Israel estimated the prevalence of IgE-mediated FA among young children at 0.85%. Although sesame was found to be a common allergen, peanuts were found to be a rare allergen. To determine the prevalence and distributions of IgE-mediated FAs among young children in Israel compared with previous data. A total of 1932 young children (56% males, 44% females) with a mean age of 22.4 months (range, 18-30 months) were sequentially recruited from 15 government family health care centers in north Israel. Parents completed a questionnaire with 2 screening questions for suspected FA. Subjects with suspected FA underwent further evaluation including telephone interview, skin prick tests, and oral food challenge as needed. After analyzing the questionnaires, 146 subjects were suspected to have FA. Seventy-nine subjects were excluded by telephone interview and 13 were excluded on the basis of negative oral food challenge. We identified 54 of 1932 (2.8%) young children with 75 IgE-mediated FAs. Thirty-nine of 54 (72.2%) had allergy to 1 food and 9 (16.6%) to 2 foods. The most common food allergens were cow's milk (1%), eggs (0.88%), sesame (0.93%), tree nuts (0.57%), peanuts (0.2%), and fish (0.2%). The prevalence of IgE-mediated FA among young children in Israel has increased dramatically from 0.85% to 2.8%. The relative prevalence of the most common food allergens is similar to that identified in 2002, with a high prevalence of sesame FA and low prevalence of peanut FA.


Dorsoventral splitting of the infundibulum in a child with pituitary hypoplasia.
Pituitary development arises from ectodermal tissue creating Rathke's pouch and ultimately the adenohypophysis anteriorly whereas neuroectodermal tissue arising from the diencephalon creates the neurohypophysis posteriorly. Alterations in pituitary development can lead to hormonal dysregulation and dysfunction. Following clinical suspicion of pituitary endocrinopathy, MRI plays a vital role in identifying and characterizing underlying structural abnormalities of the pituitary gland, as well as any associated extrapituitary findings. Here we report a case of an 18-month-old female presenting with short stature and growth hormone deficiency. MRI was notable for a shallow sella turcica, a hypoplastic adenohypophysis, thin pituitary stalk, and ectopic neurohypophysis. Interestingly, the pituitary stalk was noted to split dorsoventrally with a split pituitary bright spot and T1 hypointense lobe hypothesized to represent separation of the posterior pituitary lobes.


Bony Regrowth and New Spur Formation as Possible Causes of Failed Distal Clavicle Excision-Mid-Term Outcomes after Revision Surgery in a Matched-Pair Analysis.
Despite high rates of successful outcomes after open and arthroscopic distal clavicle excision (DCE) for symptomatic acromioclavicular joint (ACJ) degeneration, some patients present with persistent symptoms and disabilities after surgical intervention. This study aims to compare radiological, functional, and subjective outcomes of open revision surgery after failed arthroscopic DCE to primary successful arthroscopic DCE. In this retrospective case-control study, 10 patients who underwent open DCE revision were age- and gender-matched with 10 patients who did not require revision surgery after DCE. Radiographic evaluation included presence of acromioclavicular spurs and acromioclavicular joint distance. Functional and subjective outcomes were assessed using the CS, SSV, SST, VAS for pain, patient's satisfaction, ASES and quick DASH score. At the latest postoperative follow-up (case: 57.3 ± 19.2 months; control: 63.5 ± 16.3 months), spur formation was detected in twice as many cases in the revision group, while acromioclavicular distance showed no significant difference. However, a significant bony regrowth was noticed in the revision group between revision surgery and latest follow-up, with a decrease of the acromioclavicular distance from 9.2 ± 1.6 mm to 5.9 ± 4.6 mm (p = 0.026) and a development of new spur formations in 30% of cases. There were no significant differences in overall CS between the revision and control group (p = 0.174) at final follow-up, but the control group scored significantly higher in the CS subgroups pain (p = 0.012) and internal rotation (p = 0.016). Mean SSV was significantly lower in the revision (65.5 ± 22.3%) compared to the control group (85.9 ± 16.4%; p = 0.031). Bony regrowth at the distal clavicle presenting as postoperative AC-distance narrowing and new spur formation was observed more distinctly in the revision group. Despite a slight increase in postoperative outcomes after revision surgery, subjective satisfaction and recalcitrant pain remain a concern. Therapeutic Level III, retrospective case-control study.


Measured GFR in murine animal models: review on methods, techniques, and procedures.
Chronic kidney disease (CKD) is one of the most common chronic diseases worldwide, with increasing rates of morbidity and mortality. Thus, early detection is essential to prevent severe adverse events and the progression of kidney disease to an end stage. Glomerular filtration rate (GFR) is the most appropriate index to evaluate renal function in both clinical practice and basic medical research. Several animal models have been developed to understand renal disease induction and progression. Specifically, murine models are useful to study the pathogenesis of renal damage, so a reliable determination of GFR is essential to evaluate the progression of CKD. However, as in clinical practise, the estimation of GFR in murine by levels of serum/urine creatinine or cystatin-C could not be accurate and needed other more reliable methods. As an alternative, the measurement of GFR by the clearance of exogenous markers like inulin, sinistrin, 51Cr-EDTA, 99mTc-DTPA, 125I-iothalamate, or iohexol could be performed. Nevertheless, both approaches-estimation or measurement of GFR-have their limitations and a standard method for the GFR determination has not been defined. Altogether, in this review, we aim to give an overview of the current methods for GFR assessment in murine models, describing each methodology and focusing on their advantages and limitations.


Aspergillus fumigatus empyema, arthritis, and calcaneal osteomyelitis in a lung transplant patient successfully treated with posaconazole.
A 64-year-old male with Aspergillus fumigatus infection that had disseminated from the lung to the ankle and adjacent bone was treated successfully with posaconazole after therapy with itraconazole and amphotericin B lipid complex failed. Marked clinical improvement occurred within 6 weeks of initiation of posaconazole therapy; after 6 months, infection had resolved at all sites. The patient has had no recurrence of infection.


Electrocardiogram in a Young Woman With the Eisenmenger Syndrome.
A young woman with an atrial septal defect and the Eisenmenger syndrome has worsening symptoms and electrocardiographic changes of right ventricular hypertrophy five years later.


Ricinus communis L. A Value Added Crop for Remediation of Cadmium Contaminated Soil.
Heavy metal pollution of soil is a global environmental problem and therefore its remediation is of paramount importance. Cadmium (Cd) is a potential toxicant to living organisms and even at very low concentrations. This study was aimed to assess the effectiveness of Ricinus communis for remediation of Cd contaminated soils. For this, growth and biomass of R. communis and Cd accumulation, translocation and partitioning in different plant parts were investigated after 8 months of plant growth in Cd contaminated soil (17.50 mg Cd kg−1 soil). Eight months old plants stabilized 51 % Cd in its roots and rest of the metal was transferred to the stem and leaves. There were no significant differences in growth, biomass and yield between control and Cd treated plants, except fresh weight of shoots. The seed yield per plant was reduced only by 5 % of Cd contaminated plants than control. The amount of Cd translocated to the castor seeds was nominal i.e. 0.007 µg Cd g−1 seeds. The bioconcentration factor reduced significantly in shoots and seeds in comparison to roots. The data indicates that R. communis is highly tolerant to Cd contamination and can be used for remediation of heavy metal polluted sites.


New antitumor antibiotics, OS-4742 A1, A2, B1 and B2 produced by a strain of Streptomyces.
New antibiotics, OS-4742 A1, A2, B1 and B2, were isolated from the culture broth of the strain OS-4742, which was designated as Streptomyces matensis subsp. vineus. These compounds have anthracycline chromophores and sugar moieties, but do not contain nitrogen. They possess antimicrobial activities against Gram-positive bacteria and antitumor activities against S-180 solid tumor on mice.


Behavioural characterization in rats using the elevated alley Suok test.
Exploration of the long elevated alley (the rat Suok test) enables behavioural characterisation of anxiety, activity and neurological phenotypes in rats. Here we show that this new test is sensitive to different types of anxiety in rats, including drugs (pentylenetetrazole)-induced, light-induced and socially induced (encounter with an unfamiliar male) anxiety, as assessed by reduced Suok test horizontal, vertical, directed exploration and stops. High anxiety also leads to higher motor incoordination (as assessed by the number of falls and hind-paw slips), suggesting that this test may be used for combined profiling of anxiety, motor-vestibular anomalies and anxiety-induced motor incoordination in rats. This new behavioural paradigm may be widely used in neurobehavioural stress research, including modelling of stress-evoked states and pharmacological screening of psychotropic drugs.


Characterizing the Role of HMG-CoA Reductase in Aryl Hydrocarbon Receptor-Mediated Liver Injury in C57BL/6 Mice.
The aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor. The prototypical ligand of the AHR is an environmental contaminant called 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). TCDD exposure is associated with many adverse health outcomes in humans including non-alcoholic fatty liver disease (NAFLD). Previous studies suggest that AHR ligands alter cholesterol homeostasis in mice through repression of genes involved in cholesterol biosynthesis, such as Hmgcr, which encodes the rate-limiting enzyme of cholesterol biosynthesis called 3-hydroxy-3-methyl-glutaryl coenzyme A reductase (HMGCR). In this study, we sought to characterize the impact of HMGCR repression in TCDD-induced liver injury. C57BL/6 mice were exposed to TCDD in the presence or absence of simvastatin, a competitive inhibitor of HMGCR. Simvastatin exposure decreased TCDD-induced hepatic lipid accumulation in both sexes, but was most prominent in females. Simvastatin and TCDD (S + T) co-treatment increased hepatic AHR-battery gene expression and liver injury in male, but not female, mice. In addition, the S + T co-treatment led to an increase in hepatic glycogen content that coincides with heavier liver in female mice. Results from this study suggest that statins, which are amongst the most prescribed pharmaceuticals, may protect from AHR-mediated steatosis, but alter glycogen metabolism and increase the risk of TCDD-elicited liver damage in a sex-specific manner.


Skin malignancies in renal transplant recipients: a Brazilian center registry.
Renal transplant recipients have an increased risk of malignancies, especially nonmelanoma skin cancers, compared with the normal population. The aim of the present study was to analyze the incidence of skin malignancies in a setting of renal transplant recipients over 20 years follow-up. This retrospective analysis of medical records included posttransplant patients with biopsy-proven skin cancer. Recipients of pancreas kidney transplants or with suspected but not biopsy-proven skin malignancy were excluded from this series. Among 1300 renal transplant recipients from January 1984 to December 2006, 33 (2.5%) were diagnosed with skin malignancies during follow-up. The majority of patients were men (70.2%), of white race (97%), and with a mean posttransplant follow-up of 65 months. The most frequent skin cancer was squamous cell carcinoma (46.2%), in single or multiple lesions (50% each group). Basal cell carcinoma was diagnosed in seven patients; most presented as a single lesion (71.3%). Eight patients presented with more than one histologic type of skin cancer; most frequently squamous and basal cell carcinomas. Kaposi sarcoma was diagnosed in four patients, one of whom also had a basal cell carcinoma. The incidence of skin malignancies in this series was 2.5%. The most frequent tumor was squamous cell carcinoma, isolated or in association with basal cell carcinoma. An higher frequency was observed in white male patients, at a mean follow-up of 5 years posttransplantation.


CTCF functions as an insulator for somatic genes and a chromatin remodeler for pluripotency genes during reprogramming.
CTCF mediates chromatin insulation and long-distance enhancer-promoter (EP) interactions; however, little is known about how these regulatory functions are partitioned among target genes in key biological processes. Here, we show that Ctcf expression is progressively increased during induced pluripotency. In this process, CTCF first functions as a chromatin insulator responsible for direct silencing of the somatic gene expression program and, interestingly, elevated Ctcf expression next ensures chromatin accessibility and contributes to increased EP interactions for a fraction of pluripotency-associated genes. Therefore, CTCF functions in a context-specific manner to modulate the 3D genome to enable cellular reprogramming. We further discover that these context-specific CTCF functions also enlist SMARCA5, an imitation switch (ISWI) chromatin remodeler, together rewiring the epigenome to facilitate cell-fate switch. These findings reveal the dual functions of CTCF in conjunction with a key chromatin remodeler to drive reprogramming toward pluripotency.


The effect of tyrosine kinase inhibitor genistein on voltage-dependent K+ channels in rabbit coronary arterial smooth muscle cells.
We examined the effect of the protein tyrosine kinase (PTK) inhibitor, genistein on voltage-dependent K+ (Kv) channels in freshly isolated rabbit coronary arterial smooth muscle cells, using whole-cell patch clamp techniques. The amplitude of the Kv current was inhibited by genistein in a dose-dependent manner, with a Kd value of 7.51 microM. Genistein had no effect on the steady-state activation or inactivation of Kv channels. The applications of trains of pulses at 1 or 2 Hz caused a progressive increase in the genistein-blockade. Genistein produced use-dependent inhibition of the Kv currents, consistent with a slow recovery from inactivation in the presence of genistein. Daidzein and genistin, two inactive analogs of genistein, showed an inhibitory effect similar to that of genistein on Kv channels. Moreover, the absence of ATP inside the pipette did not influence the blocking effect of genistein. We suggest that genistein directly inhibited the Kv current, independently of PTK inhibition.


Perlecan/HSPG2: Signaling role of domain IV in chondrocyte clustering with implications for Schwartz-Jampel Syndrome.
Perlecan/heparan sulfate proteoglycan 2 (HSPG2), a large HSPG, is indispensable for the development of musculoskeletal tissues, where it is deposited within the pericellular matrix (PCM) surrounding chondrocytes and disappears nearly completely at the chondro-osseous junction (COJ) of developing long bones. Destruction of perlecan at the COJ converts an avascular cartilage compartment into one that permits blood vessel infiltration and osteogenesis. Mutations in perlecan are associated with chondrodysplasia with widespread musculoskeletal and joint defects. This study elucidated novel signaling roles of perlecan core protein in endochondral bone formation and chondrocyte behavior. Perlecan subdomains were tested for chondrogenic properties in ATDC5 cells, a model for early chondrogenesis. A region within domain IV of perlecan (HSPG2 IV-3) was found to promote rapid prechondrocyte clustering. Introduction of the mutation (R3452Q) associated with the human skeletal disorder Schwartz-Jampel syndrome limited HSPG2 IV-3-induced clustering. HSPG2 IV-3 activity was enhanced when thermally unfolded, likely because of increased exposure of the active motif(s). HSPG2 IV-3-induced clustering was accompanied by the deactivation of key components of the focal adhesion complex, FAK and Src, with increased messenger RNA (mRNA) levels of precartilage condensation markers Sox9 and N-cadherin ( Cdh2), and cartilage PCM components collagen II ( Col2a1) and aggrecan ( Acan). HSPG2 IV-3 reduced signaling through the ERK pathway, where loss of ERK1/2 phosphorylation coincided with reduced FoxM1 protein levels and increased mRNA levels cyclin-dependent kinase inhibitor 1C (Cdkn1c) and activating transcription factor 3 ( Atf3), reducing cell proliferation. These findings point to a critical role for perlecan domain IV in cartilage development through triggering chondrocyte condensation.


Appendectomy or not in middle-aged male with non-inflamed appendix in Amyand's hernia? Case report and literature review.
An Amyand's hernia is a rare disease where a vermiform appendix is found within an inguinal hernia sac. It is reported in the literature as having an incidence between 0.4%-1.0% of reported hernia cases. Typically, an incidental finding, Amyand's hernia is consequently found more frequently intra-operatively rather than preoperatively. This case is a recount of a 56-year-old male, who presented in an outpatient setting for the evaluation of right inguinal pain and bulge. The patient was diagnosed with a vermiform appendix within the indirect hernia. The patient underwent elective repair of his inguinal hernia via Transabdominal Preperitoneal (TAPP) approach of the hernia with avoidance of appendectomy. An Amyand's hernia presents a challenging diagnosis and the treatment algorithm is contingent on the condition of the appendix in individual cases. This case presents a Type 1 Amyand's hernia that was repaired through laparoscopic approach using prosthetic mesh. The aim of this case study highlights the approach to surgical decision making in the diagnosis and treatment of Amyand's hernia proposed in the current literature. This case presents a rare condition known as Amyand's hernia followed by a discussion on the epidemiology, diagnostic workup, and treatment options. Treatment is dependent on the state of the appendix found in the hernia sac and the clinical scenario. Comprehensive literature review shows that the true prevalence of this disease is lower than classically described and still has no clear standardized approaches.


Economic aspects of cervical cancer screening.
The results of a cost-effectiveness analysis of cervical cancer screening in The Netherlands are reported, emphasizing the analysis of the costs of screening and consequent diagnosis and treatment. Many organized screening policies are evaluated, differing in age-range and interval between screens. The cost estimates are based on organization charts, file studies and tariffs. The costs of screening itself are by far the most important cost component. Screening increases the costs of diagnosis. Costs for primary treatment only rise for large screening policies. Screening causes savings in costs of terminal treatment, but these are small compared with the costs of screening. The costs per life-year gained for the most efficient policies amount to DFL 24,000 for the policy with 7 invitations per woman in a lifetime and rise considerably in case of more than 10 invitations. Cervical cancer screening appears to be less cost-effective than breast cancer screening, but compared with other services the results are comparatively good. Implementing one of the efficient organized screening policies and discouraging spontaneous screening beyond that schedule leads to considerable savings. Moreover, many organized policies which are not efficient are still superior to spontaneous screening.


pandasGWAS: a Python package for easy retrieval of GWAS catalog data.
Since the NHGRI-EBI Catalog of human genome-wide association studies was established by NHGRI in 2008, research on it has attracted more and more researchers as the amount of data has grown rapidly. Easy-to-use, open-source, general-purpose programs for accessing the NHGRI-EBI Catalog of human genome-wide association studies are in great demand for current Python data analysis pipeline. In this work we present pandasGWAS, a Python package that provides programmatic access to the NHGRI-EBI Catalog of human genome-wide association studies. Instead of downloading all data locally, pandasGWAS queries data based on input criteria and handles paginated data gracefully. The data is then transformed into multiple associated pandas.DataFrame objects according to its hierarchical relationships, which makes it easy to integrate into current Python-based data analysis toolkits. pandasGWAS is an open-source Python package that provides the first Python client interface to the GWAS Catalog REST API. Compared with existing tools, the data structure of pandasGWAS is more consistent with the design specification of GWAS Catalog REST API, and provides many easy-to-use mathematical symbol operations.


Lysine succinylation, the metabolic bridge between cancer and immunity.
Lysine succinylation is a naturally occurring post-translational modification (PTM) that regulates the stability and function of proteins. It can be regulated by enzymes such as SIRT5 and SIRT7. Recently, the effect and significance of lysine succinylation in cancer and its implication in immunity have been extensively explored. Lysine succinylation is involved in the malignant phenotype of cancer cells. Abnormal regulation of lysine succinylation occurs in different cancers, and inhibitors targeting lysine succinylation regulatory enzymes can be used as potential anti-cancer strategies. Therefore, this review focused on the target protein lysine succinylation and its functions in cancer and immunity, in order to provide a reference for finding more potential clinical cancer targets in the future.


Relation of local platelet glycoprotein IIb/IIIa independent activation during coronary angioplasty in acute myocardial infarction to recovery of left ventricular function.
We assessed glycoprotein (GP) IIb/IIIa independent platelet activation in coronary sinus and peripheral blood from patients who underwent angioplasty for acute myocardial infarction and stable angina. Despite complete blockade of the activated GP IIb/IIIa receptor with abciximab in patients with acute myocardial infarction, unsuppressed local GP IIb/IIIa independent activation was associated with a lack of recovery of left ventricular function.


Influence of biotin intervention on glycemic control and lipid profile in patients with type 2 diabetes mellitus: A systematic review and meta-analysis.
Biotin is a water-soluble vitamin acting as a covalently bound coenzyme in regulating energy production. Previous studies have reported that biotin supplementation may influence blood glucose and lipid level in patients with type 2 diabetes mellitus (T2DM). We searched Pubmed, Embase, and Cochrane library databases up to 8th August 2022 for studies examining the effects of biotin supplementation in T2DM patients. Pooled effects were measured by weighted mean differences (WMDs) with 95% confidence intervals (CI) using random effects models. Inter-study heterogeneity was assessed and quantified. A total of five random controlled trials (RCT), involving 445 participants were included. It was suggested that biotin supplementation for 28 to 90 days significantly decreased the level of fasting blood glucose (FBG) (MD: -1.21 mmol/L, 95% CI: -2.73 to 0.31), total cholesterol (TC) (MD: -0.22 mmol/L, 95% CI: -0.25 to -0.19) and triglycerides (TG) (MD: -0.59 mmol/L, 95% CI: -1.21 to 0.03). No significant beneficial effects were observed on insulin (MD: 1.88 pmol/L 95% CI: -13.44 to 17.21). Evidence for the impact of biotin supplementation on the levels of glycated hemoglobin (HbA1c), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) and very low-density lipoprotein cholesterol (VLDL-C) was limited to draw conclusion. Biotin supplementation may decrease FBG, TC and TG levels. However, its influence on insulin is not significant and further studies on the effects of biotin on HbA1c, LDL-C, HDL-C and VLDL-C are expected.


Salt preference of congenic strains derived from the spontaneously hypertensive rat.
Previous studies using reciprocal crosses between the spontaneously hypertensive rat (SHR) and the normotensive Wistar-Kyoto (WKY) strain suggested a role for the Y chromosome in the SHR's exaggerated preference for saline solutions. We have reexamined the role of the Y chromosome in the salt preference of the SHR using a consomic strain derived from SHR and Brown Norway (BN-Lx) progenitors. We also studied congenic lines in which regions of BN-Lx chromosomes 8 and 20 had been introgressed into the SHR genome. Animals were given a choice of water and 0.9% saline to drink over a period of 7 days and their total fluid intake (TFI; water plus saline) and saline preference (proportion of the TFI taken as saline) calculated. SHR bearing the BN-Lx Y chromosome had a significantly reduced saline preference when compared to progenitor SHR. Evidence was also found for the existence of a region on chromosome 8, which influences fluid intake in the SHR. The causative genes involved in these effects however remain to be determined.


Cost drivers in endovascular pulmonary embolism interventions.
To determine the costs associated with endovascular pulmonary embolism (PE) interventions. Procedural costs were determined utilising time-driven activity-based costing (TDABC). A multidisciplinary team created process maps describing personnel, space, equipment, materials, and time required for each procedural step. Costs and capacity cost rates were determined using institutional and publicly available financial data. Process maps were developed for catheter-directed thrombolysis (CDT), ultrasound-assisted thrombolysis (USAT), pharmaco-mechanical thrombectomy (PMT), mechanical-aspiration thrombectomy (MAT), and aspiration thrombectomy (AT). Total costs were CDT $3,889, USAT $9,017.10, PMT $9,565.98, AT $12,126.42, and MAT $13,748.01. Tissue plasminogen activator costs represented 46.4% of the total materials cost for CDT, 13.1% for PMT, and 10.8% for USAT. Intensive care unit costs constitute 33.4% in CDT, 13.5% in USAT, and 13.1% in PMT of the total procedure costs. Highest total procedural costs were AT and MAT with materials cost comprising 82.6% and 80.3% of total costs, respectively. Costs were greatest with large-bore mechanical aspiration and least with catheter-directed thrombolysis using a multi-side hole infusion catheter. In the absence of a reference standard technique, physician-driven device selection can substantially impact the price of a procedure. Device choice and costs must be weighed against long-term technical and clinical success to maximise the healthcare value equation.


Understanding the Reasons Why Men and Women Do Not Donate Gametes.
The global under-supply of sperm and oocyte donors is a serious concern for assisted reproductive medicine. Research has explored self-selected populations of gamete donors and their ex-post rationalisations of why they chose to donate. However, such studies may not provide the necessary insight into why the majority of people do not donate. Utilising the unique open form responses of a large sample (n = 1035) of online survey respondents, we examine the reasons participants cite when asked: "Why haven't you donated your sperm/eggs?." We categorise these responses into four core themes (conditional willingness, barriers, unconsidered, and conscientious objector) and eleven lower-order themes. We find that, on average, women are more conditionally willing (8.2% difference; p = 0.008) to participate in gamete donation than men. We also find that women are more likely than men to justify their non-donation based on their reproductive history (21.3% difference; p = 0.000) or kin selection and inclusive fitness (5.7% difference; p = 0.008). However, compared to women, men are more likely to validate their non-donation based on sociocultural or social norms (6% difference; p = 0.000) or religion (1.7% difference; p = 0.030). That so many of our study participants report in-principal willingness for future participation in gamete donation speaks to the need for increased research on understanding non-donor population preferences, motivations, and behaviours.


Transseptal control of the difficult aortic annulus.
Successful control of the aortic root in extensive erosive endocarditis, with or without mycotic ventricular septal defect, frequently depends on secure prosthetic fixation to the interventricular septum. We describe transseptal suture fixation of aortic root prostheses through a pulmonary infundibular approach and recommend its early use to avoid difficult, often injurious, attempts at septal suture from within the left ventricular outflow tract.


Use of cell cultures for predicting the biological effects of mycotoxins.
The risk presented by mycotoxins is a toxicological problem. As the data given by physico-chemical analysis may be difficult to translate in terms of toxicity, however, especially when considering multiple contamination, we have developed a system for toxicological analysis of mycotoxins using cell cultures of different origins. The response of several cell types to a number of well defined mycotoxins was obtained in three days. This approach allowed us to: demonstrate and quantify a toxic effect, define some organ specificity related to the preferential action on a particular cell type, and detect an immunosuppressive effect. The results indicate that the system can be used for toxicological screening and that it has a predictive value for the pathological effects of tested products.


Small-for-gestation birth exerts a minor additional effect on functional impairment of the auditory brainstem in high-risk babies born at late preterm.
To address if small-for-gestational-age (SGA) significantly affects the developing auditory brainstem in late preterm babies with perinatal problems (i.e. high-risk), providing valuable information for management of such babies. SGA and appropriate-for-gestational age (AGA) babies born at 33-36weeks of gestation were studied at term using maximum length sequence brainstem auditory evoked response in response to 91-910/s clicks. Compared with AGA late preterm babies without perinatal problems (i.e. low-risk), the high-risk SGA babies manifested a significant increase in wave V latency and I-V interval at all 91-910/s clicks, and III-V interval at 455 and 910/s. The amplitude was smaller for wave I at 227 and 910/s, wave III at 910/s and wave V at 227 and 910/s. Compared with low-risk SGA babies, the high-risk SGA babies showed similarly abnormalities. Compared with high-risk AGA babies, the high-risk SGA babies manifested slightly different abnormalities. Brainstem auditory response was abnormal in high-risk SGA late preterm babies. The abnormalities, suggesting brainstem auditory impairment, were slightly different from high-risk AGA late preterm babies. SGA birth exerts a minor additional effect on the impaired auditory brainstem in high-risk babies born at late preterm. For these babies, neuroprotective measures should mainly target at associated perinatal problems, although the relatively minor adverse SGA effect cannot be ignored.


The Effects of Treatment on Body Mass Index in Giant Cell Arteritis: A Post Hoc Analysis of the GiACTA Trial.
Giant cell arteritis (GCA) requires treatment with high-dose, long-term glucocorticoids (GCs), and tocilizumab is often used early in the treatment paradigm. Weight gain, which is associated with morbidity and mortality, is a major concern for patients, though the factors that contribute to changes in body mass index (BMI) throughout the treatment of GCA are poorly understood. We analyzed GCA patients enrolled in the GiACTA (Tocilizumab in Giant Cell Arteritis) trial. We used univariable and multivariable mixed-effects modeling to examine the association between changes in BMI and cumulative GC dose, disease status at baseline (newly diagnosed versus relapsing), randomization to tocilizumab, and disease flares. A total of 250 patients were included (75% females, mean age 69 years). The mean ± SD BMI change over 52 weeks was 1.18 ± 1.98 kg/m2. On multivariable analysis, cumulative prednisone dose at 52 weeks was independently associated with BMI increase (β = 0.94 kg/m2 for 0-1 g exposure; β = 1.40 kg/m2 for ≥ 4 g exposure; p for trend < 0.001). Relapsing disease at baseline (β = - 0.42 kg/m2 compared to those with newly diagnosed disease; p = 0.002) and flares over 52 weeks in newly diagnosed patients (β = - 0.18 kg/m2 per flare; p = 0.03) were independently associated with lower BMI increase. Cumulative prednisone exposure is associated with increased BMI in GCA patients. In those with newly diagnosed disease, effective disease control regardless of the treatment used also contributes to BMI increase. Modest weight gain may be an indicator of adequate treatment response.


A Culex quinquefasciatus strain resistant to the binary toxin from Lysinibacillus sphaericus displays altered enzyme activities and energy reserves.
The resistance of a Culex quinquefasciatus strain to the binary (Bin) larvicidal toxin from Lysinibacillus sphaericus is due to the lack of expression of the toxin's receptors, the membrane-bound Cqm1 α-glucosidases. A previous transcriptomic profile of the resistant larvae showed differentially expressed genes coding Cqm1, lipases, proteases and other genes involved in lipid and carbohydrate metabolism. This study aimed to investigate the metabolic features of Bin-resistant individuals by comparing the activity of some enzymes, energy reserves, fertility and fecundity to a susceptible strain. The activity of specific enzymes was recorded in midgut samples from resistant and susceptible larvae. The amount of lipids and reducing sugars was determined for larvae and adults from both strains. Additionally, the fecundity and fertility parameters of these strains under control and stress conditions were examined. Enzyme assays showed that the esterase activities in the midgut of resistant larvae were significantly lower than susceptible ones using acetyl-, butyryl- and heptanoyl-methylumbelliferyl esthers as substrates. The α-glucosidase activity was also reduced in resistant larvae using sucrose and a synthetic substrate. No difference in protease activities as trypsins, chymotrypsins and aminopeptidases was detected between resistant and susceptible larvae. In larval and adult stages, the resistant strain showed an altered profile of energy reserves characterized by significantly reduced levels of lipids and a greater amount of reducing sugars. The fertility and fecundity of females were similar for both strains, indicating that those changes in energy reserves did not affect these reproductive parameters. Our dataset showed that Bin-resistant insects display differential metabolic features co-selected with the phenotype of resistance that can potentially have effects on mosquito fitness, in particular, due to the reduced lipid accumulation.


Endocytosis of N-acetylglucosamine-containing glycoproteins by rat fibroblasts expressing a single species of chicken liver glycoprotein receptor.
A cDNA clone for the chicken liver receptor which mediates endocytosis of glycoproteins containing terminal N-acetylglucosamine has been isolated and sequenced, confirming the previously obtained amino acid sequence of this protein (which is also known as the chicken hepatic lectin). This cDNA was introduced into Rat-1 fibroblasts and expressed using the promotor in the long terminal repeat of Moloney murine leukemia virus. Cells expressing chicken receptor were identified by screening with antireceptor antibodies followed by fluorescein-conjugated second antibodies. Receptor expressed in these cells was indistinguishable on gel electrophoresis from receptor isolated from liver. Three clonally isolated lines were examined for their ability to bind agalacto-alpha 1-acid glycoproteins at 0 degrees C and to take up and degrade this ligand at 37 degrees C. The receptor number (50,000/cell), affinity for ligand (35 nM), and uptake rate (5 molecules ligand/surface receptor/h) are similar to those previously observed for chicken hepatocytes, and for the uptake of asialoglycoproteins by rat hepatocytes and hepatoma cells. These findings indicate that the chicken receptor correctly traverses the endocytic pathway in a rat cell even though the cytoplasmic domain of this protein shows no primary structural homology with the corresponding portion of the rat liver receptor or with receptors found in fibroblasts.


[Seroprevalence of SARS-CoV-2 among children and adolescents in Germany-an overview].
SARS-CoV‑2 serologic studies complement and expand findings from confirmed COVID-19 cases through identification of undetected cases. This article summarizes previous results on SARS-CoV‑2 prevalence from seroepidemiological studies in Germany focusing on children and adolescents and complements the already existing overview on seroprevalence in adults from general population samples and especially blood donors in Germany. The results are based on an ongoing systematic search in study registries, in literature databases, of preprint publications, and of media reports of seroepidemiological studies in Germany and their results. As of 17 September 2021, we are aware of 16 German seroepidemiological studies focusing on children and adolescents. Results are available for nine of these studies. For almost all settings studied, SARS-CoV‑2 seroprevalence was well below 1% for preschool and elementary school children in the first COVID-19 wave and below 2% for adolescents. As the pandemic progressed, higher seroprevalences of up to 8% were found in elementary school children. Results of SARS-CoV‑2 antibody studies in children and adolescents in Germany are scarce so far and are based on non-representative samples at local or regional level. In future studies, it is necessary on the one hand to estimate which proportion of children and adolescents has already either had an infection or has been vaccinated. On the other hand, it is important to investigate physical and mental health impairments that occur after an infection.


The testosterone patch for women.
Has the FDA discriminated against female sexual dysfunction by not endorsing Procter & Gamble's (P&G) Intrinsa? Has the pendulum swung so far to the right that drugs for sexual medicine receive undue scrutiny? Why is testosterone replacement therapy available for men, but not for women? How should the pharmaceutical industry proceed with future studies? How should clinicians guide their patients? In this third and final perspective regarding Intrinsa, Sheryl Kingsberg, PhD, an expert on female sexual dysfunction and one of the lead investigators of the Intrinsa clinical trials, addresses these questions. This is a highly charged issue that appears to be on the agenda for the foreseeable future.


Gliotoxin contamination in and pre- and postfermented corn, sorghum and wet brewer's grains silage in Sao Paulo and Rio de Janeiro State, Brazil.
The aim of this study was to determine total fungal counts and the relative density of Aspergillus fumigatus and related species in silage samples intended for bovines before and after fermentation as well as to monitor the natural occurrence of gliotoxin in silage samples (pre- and postfermentation). The survey was performed in farms located in São Paulo and Rio de Janeiro States in Brazil. In addition, the ability of A. fumigatus strains and related species strains to produce gliotoxin was also evaluated. A total of 300 samples were taken, immediately after opening of the silo (3-5 months) and during the ensiling period. Fungal counts were done by the surface-spread method. Gliotoxin production ability of isolates and natural contamination were determined by HPLC. All postfermented samples had a total number of moulds exceeding 1 × 10(4) CFU g(-1), with Aspergillus sp. as the most prevalent genus. Frequency of strains, among A. fumigatus and related species, was able to produce gliotoxin was similar in pre- and postfermented samples, except for sorghum, which showed differences between both kinds of samples. The highest toxin levels were produced by strains isolated from postfermented samples. More than 50% of the samples showed gliotoxin contamination levels that exceeded concentrations known to induce immunosuppressive and apoptotic effects in cells. The present data suggest that care should be taken because gliotoxin contamination in feedstuffs could affect productivity and also present a health risk for herds. Gliotoxin was found at quite important concentrations levels in pre- and postfermented substrates and its presence could therefore probably affect the productivity and health of herds. Current conservation and management practices do not avoid contamination with A. fumigatus on silage. Therefore, farm workers should be adequately protected during its handling.


Interference of WEB 2086 and BN 52021 with Paf-induced effects on guinea-pig trachea.
1. The thienotriazolodiazepine WEB 2086 and the gingkolide BN52021 have been evaluated as antagonists of Paf-acether (Paf) by studying their effects on Paf-induced relaxation and Paf-induced prostaglandin E2 (PGE2) production in histamine-contracted guinea-pig tracheal preparations. 2. Relaxation induced by Paf 4 microM in histamine-contracted guinea-pig tracheal preparations was 39.67 +/- 3.5% (n = 30). At the same concentration, Paf significantly increased PGE2 production from histamine-contracted guinea-pig tracheal preparations. 3. WEB 2086 inhibited in a dose-related manner (IC50 = 21.2 nM) the relaxant effect induced by Paf and, at 1 microM, suppressed Paf-induced release of PGE2. 4. BN 52021 100 microM inhibited to about 60% Paf-induced relaxation of histamine-contracted guinea-pig tracheal preparations, but completely abolished Paf-induced increase in PGE2. 5. Both antagonists had no effects on relaxations induced by arachidonic acid 10 microM or PGE2 0.1-1 microM in histamine-contracted guinea-pig tracheal preparations. 6. The results are consistent with the presence of specific Paf receptors in guinea-pig trachea and indicate that a relaxant prostanoid, namely PGE2, at least partially mediates Paf-induced relaxation in this experimental model.


Biological relevance of inflammation and oxidative stress in the pathogenesis of arterial diseases.
Over the past three decades, age-adjusted rates of cardiovascular morbidity and mortality have fallen in the United States, but the prevalence of obesity and associated metabolic disorders has risen dramatically. Recent studies have begun to unravel the complex linkages between adipose and vascular tissues that may accelerate the development of atherosclerosis in the context of obesity. Experimental models indicate that inflammation and oxidative stress, which mutually amplify each other within the vasculature and in visceral fat, are key processes that drive the initiation, progression, and subsequent rupture of the atherosclerotic lesion. Emerging research is further elucidating the contributions made by chemokines and their receptors, adipokines, and miRNAs to arterial disease. Translation of these basic science findings to clinical applications represents a tantalizing possibility for reducing the global burden of obesity-associated atherosclerosis and other cardiovascular diseases.


Data related to cyclic deformation and fatigue behavior of direct laser deposited Ti-6Al-4V with and without heat treatment.
Data is presented describing the strain-controlled, fully-reversed uniaxial cyclic deformation and fatigue behavior of Ti-6Al-4V specimens additively manufactured via Laser Engineered Net Shaping (LENS) - a Direct Laser Deposition (DLD) process. The data was collected by performing multiple fatigue tests on specimens with various microstructural states/conditions, i.e. in their 'as-built', annealed (below the beta transus temperature), or heat treated (above the beta transus temperature) condition. Such data aids in characterizing the mechanical integrity and fatigue resistance of DLD parts. Data presented herein also allows for elucidating the strong microstructure coupling of the fatigue behavior of DLD Ti-6Al-4V, as the data trends were found to vary with material condition (i.e. as-built, annealed or heat treated) [1]. This data is of interest to the additive manufacturing and fatigue scientific communities, as well as the aerospace and biomedical industries, since additively-manufactured parts cannot be reliably deployed for public use, until their mechanical properties are understood with high certainty.


Developing structure-activity relationships from an HTS hit for inhibition of the Cks1-Skp2 protein-protein interaction.
Structure-activity relationships have been developed around 5-bromo-8-toluylsulfonamidoquinoline 1 a hit compound in an assay for the interaction of the E3 ligase Skp2 with Cks1, part of the SCF ligase complex. Disruption of this protein-protein interaction results in higher levels of CDK inhibitor p27, which can act as a tumor suppressor. The results of the SAR developed highlight the relationship between the sulfonamide and quinoline nitrogen, while also suggesting that an aryl substituent at the 5-position of the quinoline ring contributes to the potency in the interaction assay. Compounds showing potency in the interaction assay result in greater levels of p27 and have been shown to inhibit cell growth of two p27 sensitive tumor cell lines.


Role of sulfatase 2 in lipoprotein metabolism and angiogenesis.
This article summarizes the current evidence to support a role of sulfatase 2 (SULF2) in triglyceride-rich lipoprotein (TRL) metabolism and angiogenesis. Heparan sulfate proteoglycans (HSPG) are involved in the hepatic clearance of TRLs in mice and in humans. Different genetically modified mouse models have been instrumental to provide evidence that syndecan1, the core protein of HSPG, but also the degree of sulfation of the heparin sulfate chain, attached to syndecan 1, is important for hepatic TRL metabolism. Studies in humans demonstrate the regulating role of SULF2 in the hepatic uptake of TRL by HSPG and demonstrate the importance of 6-O-sulfation, modulated by SULF2, for HSPG function. The role of SULF2 in angiogenesis is illustrated by increased SULF2 mRNA expression in the stalk cells of angiogenic vascular sprouts that use fatty acids derived from TRL as a source for biomass production. Interestingly, SULF2 also interferes with HSPG-vascular endothelial growth factor binding, which impacts upon the angiogenic properties of stalk cells. SULF2 is a multifaceted protein involved in TRL homeostasis and angiogenesis. Future investigations should focus on the potential benefits of targeting SULF2 in atherosclerosis and angiogenesis.


Endonuclein is a cell cycle regulated WD-repeat protein that is up-regulated in adenocarcinoma of the pancreas.
The transcript encoding endonuclein, the human homolog of yeast PWP1, was previously found up-regulated in pancreatic cancer tissue. By immunohistochemistry we detected a ubiquitous presence in several tissues examined: skin, liver, thyroid gland, heart muscle, neurons, kidney, bladder, pancreas, adrenal gland, ovary, uterus, testis and prostate gland. We especially noticed that normal pancreatic exocrine cells exhibited low protein levels while pancreatic adenocarcinoma cells revealed high levels. We found a heterogeneous subcellular distribution, especially with varying nuclear levels. In proliferating cells endonuclein protein expression and localization was cell cycle dependent, with increasing levels and nuclear focusing during the interphase toward mitosis. Ultrastructural analysis revealed ER and nuclear localization. Endonuclein contains five WD-repeats, indicating a putative role in crucial regulatory activities in the nucleus as well as in the ER.


Microvascular engineering: Dynamic changes in microgel-entrapped vascular cells correlates with higher vasculogenic/angiogenic potential.
Successful strategies to promote neovascularization of ischemic tissues are still scarce, being a central priority in regenerative medicine. Microparticles harboring primitive vascular beds are appealing cell delivery candidates for minimally-invasive therapeutic vascularization. However, dynamic cellular alterations associated with in vitro vascular morphogenesis are still elusive. Here, bioengineered microgels guided the assembly of entrapped outgrowth endothelial cells (OEC) and mesenchymal stem cells (MSC) into cohesive vascularized microtissues. During in vitro maturation, OEC formed capillary-like networks enveloped in newly-formed extracellular matrix. Gene expression profiling showed that OEC acquired a mesenchymal-like phenotype, suggesting the occurrence of partial endothelial-to-mesenchymal transition (EndMT), while MSC remained transcriptionally stable. The secretome of entrapped cells became more pro-angiogenic, with no significant alterations of the inflammatory profile. Importantly, matured microgels showed improved cell survival/retention after transplantation in mice, with preservation of capillary-like networks and de novo formation of human vascular structures. These findings support that in vitro priming and morphogenesis of vessel-forming cells improves their vasculogenic/angiogenic potential, which is of therapeutic relevance, shedding some light on the associated mechanisms.


Coxsackieviruses A6 and A16 associated with hand, foot, and mouth disease in Vietnam, 2008-2017: Essential information for rational vaccine design.
Development of multivalent hand, foot, and mouth disease (HFMD) vaccines against enterovirus A71 (EV-A71) and several non-EV-A71 enteroviruses is needed for this life-threatening disease with a huge economic burden in Asia-Pacific countries. Comprehensive studies on the molecular epidemiology and genetic and antigenic characterization of major causative enteroviruses will provide information for rational vaccine design. Compared with molecular studies on EV-A71, that for non-EV-A71 enteroviruses remain few and limited in Vietnam. Therefore, we conducted a 10-year study on the circulation and genetic characterization of coxsackievirus A16 (CV-A16) and CV-A6 isolated from patients with HFMD in Northern Vietnam between 2008 and 2017. Enteroviruses were detected in 2228 of 3212 enrolled patients. Of the 42 serotypes assigned, 28.4% and 22.4% accounted for CV-A6 and CV-A16, being the second and the third dominant serotypes after EV-A71 (31.7%), respectively. The circulation of CV-A16 and CV-A6 showed a wide geographic distribution and distinct periodicity. Phylogenetic analyses revealed that the majority of Vietnamese CV-A6 and CV-A16 strains were located within the largest sub-genotypes or sub-genogroups. These comprised strains isolated from patients with HFMD worldwide during the past decade and the Vietnamese strains have been evolving in a manner similar to the strains circulating worldwide. Amino acid sequences of the putative functional loops on VP1 and other VPs among Vietnamese CV-A6 and CV-A16 isolates were highly conserved. Moreover, the functional loop patterns of VP1 were similar to the dominant patterns found worldwide, except for the T164K substitution on the EF loop in Vietnamese CV-A16. The findings suggest that the development of a universal HFMD vaccine, at least in Vietnam, must target CV-A6 and CV-A16 as two of the three major HFMD-causing serotypes. Vietnamese isolates or their genome sequences can be considered for rational vaccine design.


Risky Behaviours among Adolescent Students of Pokhara Valley: A School-Based Cross-sectional Survey.
Adolescence is the period of physical and mental vulnerabilities where health risk behaviours are common due to their experimental nature and decision-making power. A higher prevalence of risky behaviours among adolescents has been reported globally which resulted in adverse effects on health and wellbeing. The present study aims to explore the overall prevalence of health risk behaviours among adolescents. This study was conducted in 2014 using the WHO guideline of a Global School-Based Student Health Survey (GSHS) in the Pokhara valley of Nepal using a stratified clustered sampling technique. A descriptive study was conducted among 1190 adolescent school students of the valley. A self-administered questionnaire was used for data collection. The data were analysed using SPSS for descriptive statistics. The response rate of the participants was 95.35%. The prevalence of the current use of tobacco, alcohol, and drug was 15.9%, 17.1%, and 4.1% respectively. The mean age of first use of tobacco, alcohol, and drug was found 14.07±1.82, 14.19±1.78, and 15.52±1.61 years respectively. Two third (66.4%) of the participants reported having sexual intercourse one or two times within the last 12 months. Among them, 21.4 % reported no use of condoms during sexual intercourse. The mean age of first sexual intercourse was 14.69±1.56 years. Similarly, 42.2% reported being injured in the past 12 months of study. The study shows a higher prevalence of health risk behaviours among adolescents. Tobacco, drug, alcohol use, and risky sexual activities were initiated at a younger age.


General and Central Obesity Are Associated With Increased Severity of the VMS and Sexual Symptoms of Menopause Among Chinese Women: A Longitudinal Study.
Strong evidence has linked overweight and obesity to increased risks of cardiovascular disease and all-cause mortality in Chinese populations. Menopause is considered associated with increased obesity and central body fat distribution. However, the correlation between obesity and menopausal symptoms has not been well studied. To examine the associations between obesity or abdominal obesity and menopausal symptoms as women progressed from premenopausal to postmenopausal status. This study included 430 midlife Chinese women who had experienced natural menopause and were followed up for 10 years. Physical examinations and questionnaires should be completed annually. The questionnaires include the Menopause-Specific Quality of Life questionnaire, the Hospital Anxiety and Depression Scale, and other physical and behavioral factors. Among women who were not obese (n=345) or not abdominal obese (n=372) at baseline, 5.8% and 31.7% became obese or abdominal obese at the recent follow-up visit, respectively. Women at the recent follow-up visit had an increased body mass index (BMI) by 0.14%, and the waist-to-hip ratio (WHR) increased by 5.2% compared with the data at baseline. In multivariate analysis, more frequent hot flashes, moderate/severe bothered vasomotor symptoms (VMS), mild bothered sexual functioning, and less anxiety symptoms were significantly associated with obesity. Increasing age, moderate/severe bothered VMS, and less anxiety symptoms were independently associated with abdominal obesity. Multivariable analysis also showed that less education level is independently associated with both obesity and abdominal obesity. Our findings suggest that the proportion of obesity and abdominal obesity increased gradually during menopause. The increase of abdominal obesity is more rapidly than obesity in middle-aged women. Both obesity and abdominal obesity are related with severe or frequent VMS and anxiety symptoms in Chinese women. Although the proportion of obese women in China is lower than in western countries, the problem of abdominal obesity and related complications cannot be ignored.


Measurement of the analgesic effects of aspirin with a new experimental algesimetric procedure.
Using controlled long lasting noxious squeeze stimuli applied to the interdigital webs we have tried to develop experimental methods allowing us to measure the effects of peripherally acting analgesics. In the present double-blind cross-over study with 12 subjects we tested the effects of aspirin (1000 and 1500 mg) vs. placebo on subjective pain induced by alternately applied 12 N (Newton) and 8 N stimuli. During the sessions blood samples were taken in regular intervals to measure acetylsalicylate (ASA)- and salicylate (SA)-plasma levels. Analyses of variance were computed with several psychophysical parameters. Both the '12 N' and the '8 N' ratings discriminated between placebo and aspirin, however, only the ratings obtained from the stronger stimuli discriminated between two doses of aspirin. Subsequently we computed analyses of covariance with the ASA- and SA-plasma levels as covariates. Significant (negative) correlations of pain ratings and SA-plasma levels were found for the high dose of aspirin, but there were no significant correlations of ASA levels and ratings.


Erbu Zhuyu decoction improves endometrial angiogenesis via uterine natural killer cells and the PI3K/Akt/eNOS pathway a mouse model of embryo implantation dysfunction.
We investigated the effect of Erbu Zhuyu decoction (EBZY) on angiogenesis via uterine natural killer (uNK) cells and the PI3K/Akt/eNOS pathway in embryo implantation dysfunction (EID) mice. Pregnant mice were randomly divided into blank, model, EBZY, progynova, and aspirin groups. Uteri were excised on the 5th day of pregnancy for analysis. Mice in the model group showed pale uteri, a reduced implantation rate, and lower expression levels of phosphatidylinositol-3-kinase (PI3K), protein kinase B (Akt), endothelial nitric oxide synthase (eNOS) and nitric oxide (NO). Compared to the model group, implantation rates in the medium-dose and high-dose groups of EBZY were significantly higher (P < .05), PI3K and Akt mRNA expression levels in the low-dose group were significantly higher (P < .05, P < .01), and the expression of p-PI3K, p-Akt, and p-eNOS proteins in all treatment groups were significantly increased (P < .01, P < .05). The expression of NO was significantly increased in the low-dose and high-dose groups (P < .01, P < .05, respectively). The level of p-Akt protein in the high-dose group was significantly higher than those in the other treatment groups (P < .01, P < .05). There was no significant difference in the density of uNK cells (P > .05). EBZY facilitated embryo implantation in EID mice by enhancing endometrial angiogenesis via activation of the PI3K/Akt/eNOS pathway, at least in part. There was no evidence to indicate that EBZY could adjust the expression of uNK.


RcAlb-PepII, a synthetic small peptide bioinspired in the 2S albumin from the seed cake of Ricinus communis, is a potent antimicrobial agent against Klebsiella pneumoniae and Candida parapsilosis.
Antimicrobial peptides (AMPs) are important constituents of the innate immunity system of all living organisms. They participate in the first line of defense against invading pathogens such as viruses, bacteria, and fungi. In view of the increasing difficulties to treat infectious diseases due to the emergence of antibiotic-resistant bacterial strains, AMPs have great potential to control infectious diseases in humans and animals. In this study, two small peptides, RcAlb-PepI and RcAlb-PepII, were designed based on the primary structure of Rc-2S-Alb, a 2S albumin from the seed cake of Ricinus communis, and their antimicrobial activity assessed. RcAlb-PepII strongly inhibited the growth of Klebsiella pneumoniae and Candida parapsilosis, and induced morphological alterations in their cell surface. C. parapsilosis exposed to RcAlb-PepII presented higher cell membrane permeabilization and elevated content of reactive oxygen species. RcAlb-PepII also degraded and reduced the biofilm formation in C. parapsilosis and in K. pneumonia cells. Experimentally, RcAlb-PepII was not hemolytic and had low toxicity to mammalian cells. These are advantageous characteristics, which suggest that RcAlb-PepII is safe and apparently effective for its intended use and has great potential for the future development of an antimicrobial agent with the ability to kill or inhibit K. pneumoniae and C. parapsilosis cells.


What teachers need to know and be able to do: A view from teachers, students, and principals in the Brazilian context.
The aim of this study was to determine the main characteristics of a "good teacher" through the use of questionnaires designed to assess teaching skills and competences, considering the point of view of teachers, principals, and students. In total, 82 teachers, 14 principals, and 625 middle-school students from 5 public schools in São Paulo state participated in this study. Two questionnaires were applied, one designed for teachers and principals and the other for students. First, teachers and principals completed their specific questionnaire, after which the other questionnaire was applied to the students. Both questionnaires contained multiple choice questions related to eight distinct subsections. The questions were answered through the use of a Likert scale, varying from 1 ("totally disagree") to 5 ("totally agree"). The comparisons of the frequency of responses among all questionnaire subsections between teachers and principals were analyzed using a Chi-Square and the z-test, with P-values adjusted to the Bonferroni method. The statistical significance level was set as P < 0.05. The subsection "class atmosphere" presented the highest percentage of response "totally agree", closely followed by "professional engagement". Significant differences (P < 0.05) in responses were observed between teachers and principals for "teaching planning and practice", "use of time and material resources to develop classes", and "professional engagement" domains. In summary, it was demonstrated that some teaching characteristics might be more important than others, with some of these characteristics exhibiting significant differences between groups. Nevertheless, it is crucial to emphasize that all assessed educational domains may be recognized as critical teaching qualities, as all of them presented high levels of "totally agree" responses.


Normal tissue reactions and complications following high-energy neutron beam therapy. II: Complication rates adjusted for censoring.
A dose response analysis was performed on patients at risk for significant radiation injuries following neutron radiotherapy. Complication rates were calculated at various dose intervals using a maximum likelihood method that is formally equivalent to the product limit estimator of survival. The variance of each complication rate was used to weight a logistic regression on log dose. The treatment sites that were analyzed were head and neck, pelvis, thorax, and pancreas. Except for osteomyelitis of the mandible, complications of all types were considered collectively at each site and dose increment since there were too few complications to determine dose response functions for individual types of injuries. The head and neck was observed to be the site with the highest tolerance to radiation whereas the thorax was the most sensitive site.


Recycling of NAD(P) by multienzyme systems immobilized by microencapsulation in artificial cells.
Multistep enzyme systems can be immobilized in solution within semipermeable microcapsules. With the ability to recycle cofactors, a number of potentially useful systems have been made possible. Furthermore NAD+ can be retained inside the microcapsules by two approaches. (1) NAD+ can be linked to macromolecules such as dextran or polyethyleneimine. However, in this form, there are significant increases in steric hindrance and diffusion restrictions. (2) "Artificial cells" consisting of lipid-polyamide membrane microcapsules containing multienzyme systems, cofactors, and substrates can retain NAD+ in the free form. Analogous to the intracellular environments of red blood cells, free NAD+ in solution inside the microcapsules is effectively recycled by the multistep enzyme systems which are also in solution. Enzymes in the microcapsules are in high concentrations and in close proximity to one another. Any number and any concentration of different enzyme systems can be microencapsulated all within one artificial cell, within the limit of solubility of the total amount of enzymes. Products of sequential reactions inside the microcapsules are at much higher concentrations than outside. All these factors result in an optimal intracellular environment for multistep enzyme reactions. External substrates in the form of lipophilic or small hydrophilic molecules can equilibrate across the membrane to participate as initial substrates in the multistep reactions in the microcapsules. A number of potential applications are possible using this approach. The lipid-polyamide membrane artificial cell can also be used in basic research as a biochemical cell model for the simpler types of biological cells such as erythrocytes.


Glycated Hemoglobin, Fasting, Two-hour Post-challenge and Postprandial Glycemia in the Diagnosis and Treatment of Diabetes Mellitus: Are We Giving Them the Right Interpretation and Use?
This brief review is aimed to point out the importance of considering glycated hemoglobin, fasting blood glucose, post-glucose-load glycemia, and postprandial glycemia into an evolutive and dynamic criteria that may grant a better concept and understanding of the diagnostic and therapeutic status of individual patients with type-2 diabetes mellitus.


Shikimic acid, a mannose bioisostere, promotes hair growth with the induction of anagen hair cycle.
Shikimic acid (SA) has recently been found to be a major component of plant stem cells. The exact effects of SA on human hair follicles (HFs) is unknown. The purpose of this study was to examine the effects of SA on hair growth. We investigated the effect of SA on an in vivo C57BL/6 mouse model. We examined the expression of mannose receptor (MR), which is a known receptor of SA, in human HFs and the effect of SA on human dermal papilla cells (hDPCs), outer root sheath cells (hORSCs), and on ex vivo human hair organ culture. SA significantly prolonged anagen hair growth in the in vivo mouse model. We confirmed expression of the MR in human HFs, and that SA increased the proliferation of hDPCs and hORSCs. It was found that SA enhanced hair shaft elongation in an ex vivo human hair organ culture. SA treatment of hDPCs led to increased c-myc, hepatocyte growth factor, keratinocyte growth factor and vascular endothelial growth factor levels and upregulation of p38 MAPK and cAMP response element-binding protein levels. Our results show that SA promotes hair growth and may serve as a new therapeutic agent in the treatment of alopecia.


Predicting the pharyngeal airway space after mandibular setback surgery.
The purpose of this study is to propose a mathematical model to predict the change in pharyngeal airway space (PAS) associated with mandibular setback surgery. Twenty-three female adults, who were diagnosed as having skeletal Class III deformity, underwent mandibular setback surgery by bilateral sagittal split ramus osteotomy (BSSO) and orthodontic multibracket treatment. The subjects were assessed within 6 months before operation (T1) and 1 to 1.5 years after mandibular setback surgery (T2). The PAS change in the level of the base of the tongue was predicted. The equation is: PAS narrowing = 0.386 - 0.541 ANB (T1 - T2) + 0.253 Co-Gn (T1 - T2) - 0.098 SN-GoGn (T1 - T2), where ANB is the angle formed by the planes nasion-point A and nasion-point B, Co is condylion, Go is gonion, and Gn is gnathion. The PAS change can be predicted by the mandibular changes after setback surgery 66.2% with the regression equation using the change of the ANB angle, the total mandibular length (Co-Gn) and the mandibular plane (SN-GoGn), where SN is sella-nasion. The equation can be used to predict the change of pharyngeal airway space after mandibular setback surgery. In patients who have other risk factors, such as obesity, short neck, macroglossia, large uvula, and excessive soft tissue around the nasopharyngeal region, a mandibular setback surgery could possibly predispose to the development of sleep apnea syndrome.


MAP3K1 expression is associated with progression and poor prognosis of hormone receptor-positive, HER2-negative early-stage breast cancer.
In this study, we assessed whether the overexpression of MAP3K1 promotes the proliferation, migration, and invasion of breast cancer cells, which affect the prognosis of hormone receptor (HR)-positive, human epidermal growth factor receptor 2 (HER2)-negative early stage breast cancer. Two HR-positive, HER2-negative breast cancer cell lines (MCF7 and T-47D) overexpressing MAP3K1 were transfected with two MAP3K1 short hairpin RNA plasmids (shMAP3K1 [#3] and shMAP3K1 [#5]). The proliferation, migration, and invasion of these cells were then examined. We assessed whether shMAP3K1 affects the cell cycle, levels of downstream signaling molecules (ERK, JNK, p38 MAPK, and NF-κB), and sensitivity to chemotherapeutic and hormonal agents. To assess the anti-tumor effect of MAP3K1 knockdown in the breast cancer orthotopic model, MCF7 and T-47D cells treated with or without shMAP3K1 (#3) and shMAP3K1 (#5) were inoculated into the mammary fat pads of mice. In total, 182 patients with HR-positive, HER2-negative T1 and T2 breast cancer and 0-3 nodal metastases were included. Additionally, 73 patients with T1 and T2 breast cancer and negative nodes who received adjuvant endocrine therapy alone were selected as an independent validation cohort. In both cell lines, shMAP3K1 (#3) and shMAP3K1 (#5) significantly reduced cell growth, migration, and invasion by downregulating MMP-9 and by blocking the G2/M phase of the cell cycle and its regulatory molecule cyclin B1. Moreover, both shMAP3K1 (#3) and shMAP3K1 (#5) downregulated ERK-, JNK-, p38 MAPK-, and NF-κB-dependent gene transcription and enhanced the sensitivity of both cell lines to doxorubicin, docetaxel, and tamoxifen. We observed that both shMAP3K1 (#3) and shMAP3K1 (#5) inhibited tumor growth compared with that in the scrambled group of MCF7 and T-47D cell orthotopic tumors. Patients with MAP3K1 overexpression exhibited significantly poorer 10-year disease-free survival (DFS) (70.4% vs. 88.6%, p = 0.003) and overall survival (OS) (81.9% vs. 96.3%, p = 0.001) than those without MAP3K1 overexpression. Furthermore, phospho-ERK (p < 0.001) and phospho-JNK (p < 0.001) expressions were significantly associated with MAP3K1 expression, and both phospho-ERK and phospho-JNK expressions were significantly correlated with poor 10-year DFS and OS. These biological findings, including a significant association between DFS and OS, and the expressions of MAP3K1, phospho-ERK, and phospho-JNK were further validated in an independent cohort. Multivariate analysis identified MAP3K1 expression as an independent poor prognostic factor for DFS and OS. Our results indicate that the overexpression of MAP3K1 plays a major role in the poor prognosis of HR-positive, HER2-negative early stage breast cancer.


[Detection of postoperative lymphoedema in patients with breast cancer].
Lymphoedema is a severe postoperative complication after treatment of many malignancies. It is a pathological accumulation of extracellular water (ECW). Early diagnostic tool is needed. Multifrequency bioimpedance analysis (MFBIA) is a method for detection of changes in ECW. Prospective study. Dept. of Obstetrics and Gynaecology of the 2nd Medical Faculty, Charles University, Prague. We measured a control group of 72 women and a group of 74 patients undergoing a breast cancer surgery during 18 month after the surgery by MFBIA and circumferency. Characteristics of the patients were recorded. The detection of lymphoedema was done using MFBIA, circumferency measurement and upon the symptoms of the patients. The average age in the controll and tested group was 40,1 and 58,9 years. The average size of the tumour, grade and positivity if oestrogen receptors was 15,1 mm, 2,04 and 43%. In 23 patients (21%) complete lymphadenectomy was performed, in 51 patients (79%) a detection of sentinel lymph node was performed. Lymphoedema was detected in 8 women (11%). In these patients MFBIA detected lymphoedema 9 month earlier in total than other methods. MFBIA is a low-cost and precise method for the detection od early stage postoperative lymphoedema. We recommend to incorporate MFBIA into standard dispensatory plan of every patient combined with circumferency measurement.


Genetically determined encephalopathy: Rett syndrome.
Rett syndrome (RTT) is a severe neurodevelopmental disorder primarily affecting females that has an incidence of 1:10000 female births, one of the most common genetic causes of severe mental retardation in females. Development is apparently normal for the first 6-18 months until fine and gross motor skills and social interaction are lost, and stereotypic hand movements develop. Progression and severity of the classical form of RTT are most variable, and there are a number of atypical variants, including congenital, early onset seizure, preserved speech variant, and "forme fruste." Mutations in the X-linked gene methyl-CpG-binding protein 2 (MECP2) involve most of the classical RTT patients. Mutations in cyclin-dependent kinase like 5 (CDKL5) and FoxG1 genes have been identified in the early onset seizure and the congenital variants respectively. Management of RTT is mainly symptomatic and individualized. It focuses on optimizing each patient's abilities. A dynamic multidisciplinary approach is most effective, with specific attention given to epileptic and nonepileptic paroxysmal events, as well as scoliosis, osteoporosis, and the development of spasticity, which can have a major impact on mobility, and to the development of effective communication strategies for these severely disabled individuals.


Persistent DNA methylation changes associated with prenatal mercury exposure and cognitive performance during childhood.
Prenatal exposure to mercury, a known neurotoxic metal, is associated with lower cognitive performance during childhood. Disruption of fetal epigenetic programming could explain mercury's neurodevelopmental effects. We screened for epigenome-wide methylation differences associated with maternal prenatal blood mercury levels in 321 cord blood DNA samples and examined the persistence of these alterations during early (n = 75; 2.9-4.9 years) and mid-childhood (n = 291; 6.7-10.5 years). Among males, prenatal mercury levels were associated with lower regional cord blood DNA methylation at the Paraoxonase 1 gene (PON1) that persisted in early childhood and was attenuated in mid-childhood blood. Cord blood methylation at the PON1 locus predicted lower cognitive test scores measured during early childhood. Methylation at the PON1 locus was associated with PON1 expression in an independent set of cord blood samples. The observed persistent epigenetic disruption of the PON1 gene may modulate mercury toxicity in humans and might serve as a biomarker of exposure and disease susceptibility.


Theoretical insight on electronic structure and photophysical properties of a series of cyclometalated iridium(III) complexes bearing the substituted phenylpyrazole with different electron-donating or electron-accepting groups.
By using the density functional theory (DFT) and time-dependent density functional theory (TDDFT), the electronic structure and photophysical properties of a series of cyclometalated iridium(III) complexes bearing the substituted phenylpyrazole have been theoretically investigated. All studied iridium(III) complexes have the distorted octahedral geometry with cis-C,C, cis-O,O, and trans-N,N chelate disposition. The lowest lying singlet → singlet absorptions of all studied iridium(III) complexes are respectively located at 405 nm, 387 nm, 382 nm, 370 nm, and 387 nm. The calculated emission wavelengths for all studied iridium(III) complexes are 654 nm, 513 nm, 506 nm, 505 nm and 499 nm, respectively. The calculated emission wavelength for complex 4 at the CAM-B3LYP level is in good agreement with the experimental value. From the theoretical results, it can be seen that the electron-donating substituent groups have the important effect on the electronic structure and photophysical properties of all studied iridium(III) complexes. We hope that this study can provide valuable guidance for the design of new phosphorescent organic light-emitting diodes (OLEDs) materials.


Seawall-induced impacts on large river delta wetlands and blue carbon storage under sea level rise.
Coastal wetlands have been enclosed by thousands of kilometers of seawalls in China to obtain extra land for rapid socio-economic development in the coastal region. Although understanding seawall-induced impacts on delta wetlands and their ecosystem can provide valuable decision-making information to support coastal management, quantifying and measuring long-term, cumulative ecological impacts of harden seawall under sea level rise (SLR) remains a vital research gap. In this study, by combining the land-use transformation trajectory analysis, ecosystem services assessment, and the SLAMM (Sea Level Affecting Marshes Model), we have explored the seawall-induced effects on temporal-spatial dynamics of tidal wetlands and the Coastal Blue Carbon storage (CBCs) in the Yellow River Delta (YRD) under the SLR by 2050 and 2100. Our study revealed that the delta wetland area would have increased by 2327.87 km2 after seawall removal without regard for SLR while increasing by 3050 km2 in 2100 in both seawall scenarios under SLR. The effects of driving processes trajectory on the changes in CBCs indicated two-sided seawall-induced impacts on the delta wetlands in the YRD, i.e., functioning as a physical coastal defense to prevent coastal erosion (before 2050) while intensifying coastal squeeze effects and quickening the loss in delta wetlands and the CBCs by hindering their inland migration under SLR. For example, the gap of CBCs between the seawall-impacting and seawall-removal scenarios would have reached at 9.94 × 106 Mg by 2050 under the SLR, and the magnitude of the final decrease effect on CBCs induced by the seawall-impacting would be nearly 5 times higher than its gain after seawall-removal in the regressive succession, while the same magnitudes in the salinization process on both scenarios. Our study has provided valuable insights for shoreline management by mitigating seawall-induced impacts on the delta wetlands and their ecosystem services such as CBCs.