Ageing, exercise and food restriction: effects on body composition.
This study was undertaken to reevaluate the effect of ageing on body composition and to compare the effects of exercise and food restriction on body fat content in rats. Body fat content increased progressively between ages 12 and 28 months in sedentary freely eating male Long-Evans rats. A program of 3 h of swimming/day caused body weight to plateau at a level approximately 20% below that of the freely eating sedentary rats, primarily due to a more than 100 g lower body fat content in the swimmers. Lean body mass and protein content decreased progressively with ageing; in the sedentary animals, this decrease was significant by 18 months. The swimming protected against the decline in lean body mass and protein content up to age 18 months, but did not prevent a large decrease in lean body mass between ages 18 and 24 months. Food restriction that kept the sedentary rats at the same weight as the swimmers had its major effect on body fat content, which was approximately 40% lower than in the sedentary freely eating rats at all ages. The differences in body composition between the swimmers and food restricted sedentary animals were relatively small and consisted primarily of a lower body fat content in the swimmers. In conclusion, our results show that lean body mass declines with ageing in rats and that this loss of lean tissue is counterbalanced by an increase in fat. Exercise is highly effective in minimizing the increase in body fat, and may delay the decline in lean body mass, with ageing.


Self-Care Adherence And Barriers To Good Glycaemic Control In Nepalese Type 2 Diabetes Mellitus Patients: A Hospital-Based Cross-Sectional Study.
The patient believes in adherence to medication rather than to self-care adherence and lifestyle changes for the management of diabetes. This study was carried out to establish the association of self-care adherence and their barriers in poor glycemic control in our diabetic population. This cross-sectional study was conducted among 480 already diagnosed diabetes outpatients attended in our two hospitals. Glycaemic control was defined by levels of HbA1c. Socio-demographic data, lifestyle variables and anthropometric measurements were recorded using a standard questionnaire. Fasting blood glucose, HbA1c and lipid profiles were estimated using the manufacturer's guideline. Student's t-test and one-way ANOVA were used for comparison between different groups and the correlation was established by Spearman correlation. Risk factors associated with poor glycaemic control were verified by logistic regression analysis. The mean HbA1c of the study population was 7.4±1.3% and 65.4% had poor glycaemic control with mean 8.0±1.1%. Higher HbA1c levels were significantly associated with duration of diabetes, a number of drugs used, patient-physician relationship and knowledge about diabetes. The poor glycaemic control was significantly associated with low adherence of following the meal plan, regular medication and regular exercising (p<0.001). Among all the barriers, a too busy schedule for following the meal plan, taking medications and exercising regularly was significantly correlated with HbA1c levels. Multivariable logistic regression analysis showed irregular meal plan (OR=5.27), irregular exercise (OR=2.25), number of medication used (OR= 0.19) and lesser extent patient-physician relationship (OR=2.68) were independent risk factors for poor glycaemic control. The poor glycaemic control was associated with poor adherence to self-care adherence and their barriers in our diabetic population. Integrated knowledge on diabetes management should be targeted to improve glycaemic control in our communities.


First mitochondrial genome of Euprosopia sp. (Diptera: Platystomatidae).
Platystomatid flies, one of the largest families in Diptera. Here, we determined the complete mitogenome of Euprosopia sp., which is the first for the family Platystomatidae. The 16,266 base pair (bp) mitogenome comprises of 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), 2 ribosomal RNA genes, and a control region. The gene order and the orientation are similar to those of other sequenced Acalyptratae species except tRNA Ser ( AGN ) gene were substituted by another tRNA Phe gene. All PCGs start with ATN codons, except COI and ND1, and end with TAA or incomplete stop codon TA + tRNA. The 22 tRNAs have a typical cloverleaf secondary structure. Phylogenetic analyses base on 13 Diptera species supported the monophyly of Superfamily Tephritoidea.


HLA-DPB1 mismatching results in the generation of a full repertoire of HLA-DPB1-specific CD4+ T cell responses showing immunogenicity of all HLA-DPB1 alleles.
Clinical studies have indicated that HLA-DPB1 functions as a classical transplantation antigen in allogeneic stem cell transplantation. Mismatching for HLA-DPB1 was associated with an increased risk of graft-versus-host disease (GVHD), but also a decreased risk of disease relapse. However, specific HLA-DPB1 mismatches were associated with poor clinical outcome. It was suggested that this unfavorable effect was caused by a difference in immunogenicity between HLA-DPB1 alleles. To analyze whether immunogenicity of HLA-DPB1 mismatches could be predicted based on the presence or absence of specific amino acid sequences we developed a model to generate allo-HLA-DPB1 responses in vitro. We tested in total 48 different stimulator/responder combinations by stimulating CD4(+) T cells from 5 HLA-DPB1 homozygous individuals with the same antigen-presenting cells transduced with different allo-HLA-DPB1 molecules. HLA-DPB1 molecules used for stimulation comprised 76% to 99% of HLA-DPB1 molecules present in different ethnic populations. We show that all HLA-DPB1 mismatches as defined by allele typing resulted in high-frequency immune responses. Furthermore, we show that crossrecognition of different HLA-DPB1 molecules is a broadly observed phenomenon. We confirm previously described patterns in crossrecognition, and demonstrate that a high degree in similarity between HLA-DPB1 molecules is predictive for crossrecognition, but not for immunogenicity.


Respiration, predatory behavior and prey consumption by Podisus nigrispinus (Heteroptera: Pentatomidae) nymphs exposed to some insecticides.
Podisus nigrispinus Dallas (Heteroptera: Pentatomidae) preys on insect pests in eucalyptus plantations where it can be exposed to insecticides used in pest control. The effect of insecticides on non-target natural enemies requires further study. The objective of the present study was to evaluate the side-effects of Bacillus thuringiensis (Bt), permethrin, tebufenozide and thiamethoxam on third instar nymphs of the predator P. nigrispinus in the laboratory. The toxicity of insecticides for this insect was determined by estimating their lethal concentrations. Podisus nigrispinus behavior after exposure to insecticides was analyzed using a video tracking system and the respiratory rate with a respirometer. Prey/nymph consumption was assessed after 24 h of starvation. The preference of P. nigrispinus nymphs, for prey treated or not with the insecticides, was evaluated in free choice tests. The insecticides Bt [LC50 = 1.10(0.83-1.46) mg mL-1], permethrin [LC50 = 0.25(0.17-0.34) mg mL-1], tebufenozide [LC50 = 5.71(4.17-7.57) mg mL-1] and thiamethoxam [LC50 = 0.04(0.02-0.06) mg mL-1] are toxic to P. nigrispinus nymphs. Bt and the insecticides tebufenozide, permethrin and thiamethoxam reduced the respiratory rate of P. nigrispinus. The insecticides permethrin, tebufenozide and thiamethoxam affect the locomotion of this insect's nymphs. Prey treated with Bt, permethrin and thiamethoxam are less preferred by P. nigrispinus. The survival of the nymphs of this predator was 93.3%, 66.7%, 56.6%, 0% and 0% in the control, tebufenozide, Bt, permethrin and thiamethoxam treatments, respectively. In addition, the reduction of prey consumption, treated with neurotoxic insecticides, reduces the predatory potential of this natural enemy. Bt and tefubenozide present low toxicity for P. nigrispinus, but the neurotoxic products have low compatibility with this natural enemy and, therefore, are not recommended, with this predator in the management of forest insect pests.


A frameshift mutation results in a truncated nonfunctional carboxyl-terminal pro alpha 1(I) propeptide of type I collagen in osteogenesis imperfecta.
A codon frameshift mutation caused by a single base (U) insertion after base pair 4088 of prepro alpha 1(I) mRNA of type I procollagen was identified in a baby with lethal perinatal osteogenesis imperfecta. The mutation was identified in fibroblast RNA by a new method that allows the direct detection of mismatched bases by chemical modification and cleavage in heteroduplexes formed between mRNA and control cDNA probes. The region of mismatches was specifically amplified by the polymerase chain reaction and sequenced. The heterozygous mutation in the amplified cDNA most likely resulted from a T insertion in exon 49 of COL1A1. The frameshift resulted in a truncated pro alpha 1(I) carboxyl-terminal propeptide in which the amino acid sequence was abnormal from Val1146 to the carboxyl terminus. The propeptide lacked Asn1187, which normally carries an N-linked oligosaccharide unit, and was more basic than the normal propeptide. The distribution of cysteines was altered and the mutant propeptide was unable to form normal interchain disulfide bonds. Some of the mutant pro alpha 1(I)' chains were incorporated into type I procollagen molecules but resulted in abnormal helix formation with over-hydroxylation of lysine residues, increased degradation, and poor secretion. Only normal type I collagen was incorporated into the extracellular matrix in vivo resulting in a tissue type I collagen content approximately 20% of that of control (Bateman, J. F., Chan, D., Mascara, T., Rogers, J. G., and Cole, W. G. (1986) Biochem. J. 240, 699-708).


Selective depression of the C4 component of complement: evidence for an association with genetic deficiency of C4 and dermatologic diseases.
Thirty-four patients with selective low serum C4 levels were found among 1,731 patients studied in a hospital-based routine clinical immunology laboratory. A high incidence of dermatologic diseases, particularly discoid lupus erythematosus and angioedema/urticaria, was present among patients with selective low C4. Although evidence for activation of C4 was present in patients' sera, studies to determine the presence of known activation mechanisms of complement were negative. Genetic C4 typing in four pedigrees showed that all propositi carried the null allele B*QO. It is postulated that the selective C4 depression in these patients is related to the skin lesions and this partial genetic deficiency.


General surgery training in Spain: core curriculum and specific areas of training.
The royal decree RD 639/2014 has been published, regulating among others, the core curriculum, and specific areas of training (SAT). It is of great interest for the specialty of General and Digestive Surgery (GS and DS). The aim is to expose and clarify the main provisions and reflect on their implications for the practical application of the core curriculum and SAT in the specialty of General and Digestive Surgery, to promote initiatives and regulations. This RD will be a milestone in our specialty that will test the strength of the specialty, if it does not finally culminate in its degradation against the emergence of new surgical specialties. A new stage begins in which the Spanish Association of Surgeons should be involved to define the conceptual basis of GS and DS in the XXI century, and the creation of new SAT to continue to maintain the "essence of our specialty".


Delayed dermal hypersensitivity in rhesus monkeys (Macaca mulatta) immunized against Plasmodium knowlesi.
Eighteen Rhesus monkeys were immunized with a fresh or lyophilized antigen preparation obtained from erythrocytes infected with Plasmodium knowlesi. Sixteen of these monkeys showed a positive delayed hypersensitivity response when subsequently skin tested with the immunizing antigen. Ten of the 16 positive reactors survived a P, knowlesi challenge, the remaining 6 showed several indications of protection. None of the 18 control monkeys gave a positive skin reaction or survived the challenging inoculum. Macro- and microscopical evidence suggests that the dermal reaction is of a delayed type. Preliminary biochemical analysis of the antigen used in this experiment is included and its significance is discussed. Hematological data is also presented.


Initial prescription of antipsychotics and antidepressants in general practice and specialist care in Norway.
To examine the incidence rates of antipsychotic (AP) and antidepressant (AD) drug treatment in Norway and the proportions initiated in general practice and specialist care respectively. Data on all prescriptions of APs and ADs dispensed to the general population in Norway from 1 January 2004 until 31 August 2009 were extracted from the Norwegian Prescription Database. This information was merged with data about general practitioners (GPs) from the Norwegian Regular General Practitioner Scheme. One-year incidence rates per 1000 inhabitants were 3.4 for APs and 8.6 for ADs. GPs initiated 58% of APs and 73% of ADs, while psychiatrists initiated 15% and 6% respectively. Psychiatrists initiated treatment more often among younger patients, and they prescribed relatively newer drugs more commonly than GPs. A large share of incident users did not refill their prescriptions for APs (57%) or ADs (33%). GPs have a key role as regards initiating treatment with APs and ADs in Norway, while psychiatrists' influence seems limited, particularly among older patients. Efforts for quality improvement of mental health care need to involve primary health care. In addition, an increased focus from psychiatrists towards the increasingly ageing part of the population seems requisite.


Effects of comorbid alcohol use disorder on the clinical outcomes of first-episode schizophrenia: a nationwide population-based study.
Alcohol use disorder (AUD) is a common psychiatric comorbidity in schizophrenia, associated with poor clinical outcomes and medication noncompliance. Most previous studies on the effect of alcohol use in patients with schizophrenia had limitations of small sample size or a cross-sectional design. Therefore, we used a nationwide population database to investigate the impact of AUD on clinical outcomes of schizophrenia. Data from the Health Insurance Review Agency database in South Korea from January 1, 2007 to December 31, 2016 were used. Among 64,442 patients with first-episode schizophrenia, 1598 patients with comorbid AUD were selected based on the diagnostic code F10. We performed between- and within-group analyses to compare the rates of psychiatric admissions and emergency room (ER) visits, and medication possession ratio (MPR) between the patients with comorbid AUD and control patients matched for the onset age, sex, and observation period. The rates of psychiatric admissions and ER visits in both groups decreased after the time point of diagnosis of AUD; however, the decrease was significantly greater in the patients with comorbid AUD compared to the control patients. While the comorbid AUD group showed an increase in MPR after the diagnosis of AUD, MPR decreased in the control group. The rates of psychiatric admissions, ER visits, and MPR were worse in the comorbid AUD group both before and after the diagnosis of AUD. The results emphasize an importance of psychiatric comorbidities, especially AUD, in first-episode schizophrenia and the necessity of further research for confirmative findings of the association of AUD with clinical outcomes of schizophrenia.


Postischemic hyperglycemia worsens neurologic outcome after spinal cord ischemia.
The neurologic effect of induced hyperglycemia in the postischemic period was investigated with a rat aortic occlusion model. Sprague-Dawley rats weighing 200 to 350 gm were anesthetized, intubated, and ventilated with 1% to 1.5% halothane. Temperature was continuously monitored and maintained at 37 degrees +/- 0.5 degrees C. The chest was opened, the thymus excised, and the aortic arch exposed. Snares were placed around the aorta distal to the left subclavian artery and the right and left subclavian arteries. The three vessels thus isolated were occluded for 8 minutes. With snare release and withdrawal, the rats received an intraperitoneal injection of 5% dextrose in water (2 gm/kg) or an equivalent volume of 0.9% saline solution. In a second group of rats the administration of glucose or saline solution was delayed until 30 minutes after snare release. Blood samples for blood glucose determination were obtained before operation, before occlusion, immediately after occlusion, and 15, 30, 45, 60, and 240 minutes after occlusion. A neurologic deficit score was assigned at 1, 4, 18, and 24 hours after occlusion to quantify hindlimb neurologic deficit based on 15-point scale (0 = normal, 15 = severe deficit). Sham-operated rats received the same operation and injection, but the snares were only manipulated and not made occlusive. The rats that were administered glucose immediately after snare release showed a statistically significant exacerbation of lower extremity neurologic deficit at 24 hours after occlusion (p < or = 0.05, Mann-Whitney U test). The sham-operated rats were normal (0 score) at 24 hours. Significant elevation of blood glucose (321 +/- 33 mg/dl) was seen in the glucose-injected rats at 15 minutes and continued for up to 4 hours after occlusion (p = 0.040 and 0.014, respectively; Student's t test). Postischemic hyperglycemia immediately after a standard spinal cord ischemic stress worsens neurologic outcome.


Comparative activity of plazomicin against extended-spectrum cephalosporin-resistant Escherichia coli clinical isolates (2012-2017) in relation to phylogenetic background, sequence type 131 subclones, blaCTX-M genotype, and resistance to comparator agents.
Extended-spectrum cephalosporin-resistant Escherichia coli (ESCREC) are a growing threat. Leading ESCREC lineages include sequence type ST131, especially its (blaCTX-M-15-associated) H30Rx subclone and (blaCTX-M-27-associated) C1-M27 subset within the H30R1 subclone. The comparative activity against such strains of alternative antimicrobial agents, including the recently developed aminoglycoside plazomicin, is undefined, so was investigated here. We assessed plazomicin and 11 comparators for activity against 216 well-characterized ESCREC isolates (Minnesota, 2012-2017) and then compared broth microdilution MICs with phylogenetic and clonal background, beta-lactamase genotype (blaCTX-M; group 1 and 9 variants), and co-resistance. Percent susceptible was > 99% for plazomicin, meropenem, imipenem, and tigecycline; 96-98% for amikacin and ertapenem; and ≤ 75% for the remaining comparators. For most comparators, MICs varied significantly in relation to multiple bacterial characteristics, in agent-specific patterns. By contrast, for plazomicin, the only bacterial characteristic significantly associated with MICs was ST131 subclone: plazomicin MICs were lowest among O16 ST131 isolates and highest among ST131-H30R1 C1-M27 subclone isolates. Additionally, plazomicin MICs varied significantly in relation to resistance vs. susceptibility to comparator agents only for amikacin and levofloxacin. For most study agents, antimicrobial activity against ESCREC varied extensively in relation to multiple bacterial characteristics, including clonal background, whereas for plazomicin, it varied only by ST131 subclone (C1-M27 isolates least susceptible, O16 isolates most susceptible). These findings support plazomicin as a reliable alternative for treating ESCREC infections and urge continued attention to the C1-M27 ST131 subclone.


Three-dimensional structure of apotransketolase. Flexible loops at the active site enable cofactor binding.
The structure determination of apotransketolase and the comparison of its three-dimensional structure with that of the holoenzyme has revealed that no large conformational changes are associated with cofactor binding. Two loops at the active site are flexible in the apoenzyme which enables ThDP to reach its binding site. Binding of the cofactor induces defined conformations for these two loops at the active site. One of these loops is directly involving in binding of the cofactors, Ca2+ and ThDP. This loop acts like a flap which closes off the diphosphate binding site. After binding of the cofactor, residues of this loop form interactions to residues of loop 383-398 from the second subunit. These interactions stabilize the conformation of the two loops from a flexible to a 'closed' conformation.


Polyacrylamide-alginate (PAAm-Alg) and phospho-L-tyrosine-linked PAAm-Alg monolithic cryogels: Purification of IgG from human serum.
In recent decades cryogels as monolithic materials have gained interest as stationary phase in chromatography for purification of biomolecules. In this study, polyacrylamide-alginate (PAAm-Alg) monolithic cryogels were prepared by cryo-copolymerization of acrylamide and alginate monomers and methylene-bisacrylamide as crosslinker to be used as a matrix in affinity chromatography for purification of proteins. Ortho-phospho-L-tyrosine (P-Tyr) was covalently attached onto PAAm-Alg cryogels via bisoxirane-activation (PAAm-Alg-Bix-P-Tyr) and both derivatized and non-derivatized cryogels were utilized for the purification of immunoglobulin G (IgG) from human serum. Cryogels were characterized by scanning electron microscopy, swelling tests, elemental analysis, FTIR, and flow dynamics. The effects of buffer systems, conductivity, and pH on IgG adsorption were studied. Through breakthrough curve analysis a dynamic capacity of 9.2 mg IgG/mL with an IgG purity of 94% was obtained (based on ELISA analysis of IgG and albumin) for PAAm-Alg-Bix-P-Tyr cryogel when human serum was diluted in 10 mmol/L NaP buffer at pH 6.0. The adsorption isotherm data were well described by the Langmuir model with value of maximum adsorption capacity of 36.12 ± 3.63 mg of IgG/g for PAAm-Alg-Bix-P-Tyr. The PAAm-Alg-Bix-P-Tyr cryogel provides an attractive alternative for adsorption of IgG from human serum.


Potent synergistic combination of rosuvastatin and levofloxacin against Staphylococcus aureus: in vitro and in vivo study.
The present study aims to evaluate the capability of rosuvastatin to synergize with levofloxacin against Staphylococcus aureus. Rosuvastatin inhibited the growth of S. aureus with minimum inhibitory concentration of 16 μg ml-1 . Additionally, it showed a bactericidal effect at 4x minimum inhibition concentration. Using a checkerboard method, a synergistic effect was recorded when rosuvastatin was combined with levofloxacin showing against S. aureus isolate 28 (S 28). Furthermore, this combination was also able to display a significant reduction in biofilm formation (92·8%) and suppress the production of coagulase and β-haemolysin, and virulence factors of S. aureus isolate 28. An animal model for wound infection was used to assess the therapeutic effect of the test combination, in vivo. It was found that the test combination reduced the bacterial burden in the infected wounds by 91·3%. Pathological and histological analyses have revealed a decline in cell infiltration in the excisional wound skin tissue after treatment with rosuvastatin and levofloxacin combination. Rosuvastatin combined with levofloxacin can be considered as a promising solution to combat S. aureus antibiotic resistance phenomenon. This study unveils the potential effect of rosuvastatin when used in combination with levofloxacin can be used as a topical antibacterial agent to treat S. aureus skin infections.


Enhanced feeding in very-low-birth-weight infants may cause electrolyte disturbances and septicemia--a randomized, controlled trial.
High supply of protein and energy has been introduced to very-low-birth-weight infants to improve growth and cognitive development. The aim of this study was to compare two different feeding strategies on postnatal growth and clinical outcome during neonatal hospitalization. Fifty very-low-birth-weight infants were randomized to either an enhanced or a standard feeding protocol within 24 h after birth. Chi-square and T-tests were applied. First week protein, fat and energy supply was significantly higher in the intervention group compared to the control group (all P < 0.001). After inclusion of 50 patients we observed a higher occurrence of septicemia in the intervention group, 63% vs. 29% (P = 0.02), and no more patients were included. The infants in the intervention group demonstrated improved postnatal growth, but they also disclosed significant electrolyte deviations during the first week of life with hypophosphatemia, hypokalemia and hypercalcemia. First week phosphate nadir was lower in the infants experiencing septicemia (1.23 (0.50) mmol/L) as compared to the infants without (1.61 (0.61) mmol/L) (P = 0.03). Our study implies that enhanced feeding may induce electrolyte imbalances in VLBW infants, and that deleterious side effects similar to those seen in refeeding syndrome may occur. ClinicalTrials.gov, number NCT01103219 and the EudraCT number is 2010-020464-38.


Bronchoscopic interventions for plastic bronchitis in children without structural heart disease.
Plastic bronchitis (PB) is a rare and life-threatening complication encountered in several disease states that leads to airway obstruction by branching casts. PB is most often reported in children with cyanotic congenital heart disease where recurrence is common, and mortality is high. There is limited data on optimal management strategies or recurrence of non-structural heart disease-related PB in children. We describe the clinical features, management, and outcomes in our cohort of children with non-structural heart disease-related PB. Among the 12 identified patients, asthma was the most common (67%) diagnosis. Ventilatory requirements ranged from room air to one patient who required extracorporeal membrane oxygenation (ECMO). Most patients (92%) required bronchoscopy, and cryotherapy was successfully utilized in two patients to relieve refractory obstructive airway casts. All patients received chest physiotherapy, and 11 patients were treated with two or more medications. There was one mortality despite ECMO, and one-third had recurrent PB, all of whom had asthma.Conclusion: Asthma is a risk factor for recurrent PB. Bronchoscopic interventions including cryotherapy are safe and effective treatment options in patients with refractory PB. What is Known: • Plastic bronchitis is a rare but life-threatening cause of airway obstruction caused by branching casts that are generally reported in patients with congenital heart disease. What is New: • In children without structural heart disease, asthma is a risk factor for recurrent plastic bronchitis. Cryotherapy via bronchoscopy is a safe and effective intervention in patients with refractory plastic bronchitis.


Mutual interaction between glycation and oxidation during non-enzymatic protein modification.
Aging pathogenesis involves non-enzymatic modifications of proteins; protein oxidation, glycation and their interactions have aroused a particular interest. Possible interrelations between oxidation and glycation have been evaluated in vitro: bovine serum albumin was oxidized by gamma-irradiation and then exposed to in vitro glycation. Fluorescence modifications induced by radiolytic oxidation and glycation were similar and tended to be additive. Both non-enzymatic processes provoked a loss of free sulfhydryl groups and a strong increment of protein carbonyl content: this supports that glycation can act through oxidative mechanisms. The observed rearrangement of amino groups after irradiation could predispose proteins to glycation attacks. Protein peroxides generated during irradiation appear able to give birth to further protein modifications leading to the generation of carbonyl groups and to interact with monosaccharides, probably stimulating their autoxidation and in turn glycative protein damage. Glycation increases the oxidation-mediated structural damage revealed by SDS-PAGE. Therefore our data support the hypothesis of mutual enhancement between oxidation and glycation of proteins and suggest possible molecular mechanisms of interactions.


Vaginal cuff brachytherapy in the adjuvant setting for patients with high-risk early-stage cervical cancer.
To evaluate local control and survival of high-risk patients with early-stage cervical cancer submitted or not to vaginal cuff brachytherapy in the postoperative setting. In this retrospective cohort of patients treated from 2010 to 2017, patients were eligible if they had confirmed histological diagnosis of cervical cancer treated with surgery and adjuvant radiotherapy with or without chemotherapy. Vaginal cuff brachytherapy (VCB) was indicated according to the radiation oncologist discretion. Seventy-nine patients were selected, with a median age at diagnosis of 47.5 years (26-77). Brachytherapy was delivered to 59 patients (74.7%). There were no significant differences between the VCB and the no-VCB groups. A total of 13 (16.5%) patients presented one or more events, 5 (25%) and 8 (13.5%) events in the no-VCB and VCB group, respectively. Most recurrences were pelvic and/or vaginal: 7/20 (35%) in the no-VCB group and 9/59 (10.2%) in the VCB group. There were eight systemic relapses with eight deaths. With a median followup of 45 months, mean overall survival and disease-free survival were, respectively, 85.1 and 83.8 months. No variables were correlated with overall survival. The only factor positively correlated to disease-free survival was VCB, with a mean of 86.9 and 68.4 months for patients who did and did not receive brachytherapy, respectively (p = 0.043). Vaginal recurrence was lower in the brachytherapy group, but with no statistical significance (p = 0.065). VCB was associated with a reduced recurrence rate in the postoperative setting of high-risk patients with early-stage cervical cancer.


Polyandrous Mexican Fruit Flies: Second Male Paternity and Biological Attributes of Transgenic Strains.
Anastrepha ludens (Diptera: Tephritidae), is a damaging agricultural pest. Currently, the Sterile Insect Technique (SIT) is used as part of its control. The SIT consists of the mass-rearing, sterilization, and release of insects in target areas. Sterile males mate with wild females, and prevent them from laying fertile eggs. However, even if females mate with sterile males, they can then remate with a second male. If this second male is wild, then this could reduce the efficiency of the SIT by producing viable offspring. The amount of progeny produced by second males (P2 values) for A. ludens is unknown. Here, we evaluated the biological attributes, mating competitiveness, and the proportion of male paternity gained by the second male, using strains that carry fluorescent marker genes and can be potentially used to develop transgenic sexing strains. Furthermore, the transgenic strains were irradiated, to test their ability to induce sterility in females. We found that the 443-G strain had significantly higher larval survival than the 419-R strain. No significant difference was found between the two strains in their mating probability with wild females. We found P2 values between 67 and 74% for the 419-R and the 443-G strain, respectively. Second male sperm precedence only decreased slightly after 12 days, suggesting that sperm from the first and second male is not mixing with time, but rather the second male's sperm prevails. Furthermore, sterile 443-G males induced significantly higher sterility in females than sterile males from the 419-R strain. The apparent lower ability of the 443-G strain to inhibit female remating should be further investigated. Knowledge of the pre and postcopulatory performance of transgenic strains will help in understanding their potential for control.


Molecular and MALDI-ToF MS differentiation and antifungal susceptibility of prevalent clinical Fusarium species in China.
Fusarium species are emerging causative agents of superficial and disseminated human infections. Early diagnosis and treatment contribute to better prognosis of severe infection. To detect the effectiveness of matrix-assisted laser desorption ionisation time of flight mass spectrometry (MALDI-ToF MS) for Fusarium identification, and evaluate the susceptibility profiles to clinical available antifungals. All 203 clinical Fusarium isolates and 25 environmental isolates were identified by using translation elongation factor 1-alpha (TEF1) and RNA polymerase subunit II (RPB2) sequencing and MALDI-ToF MS. Antifungal susceptibility testing was determined by a microdilution method following the CLSI approved standard M38-A3 document. Correct identification rates at the species and genus levels were 89.04% (203/228) and 95.18% (217/228), respectively, using Bruker Filamentous Fungi Library 1.0 combined with the novel database. Seven species complexes with 19 Fusarium species were identified, including F. solani (59.21%, n = 135), F. verticillioides (17.54%, n = 40), F. proliferatum (6.58%, n = 15) and F. oxysporum (4.39%, n = 10). Four uncommon species complexes (F. incarnatum-equiseti SC, F. dimerum SC, F. redolens SC and F. sporotrichioides SC) were also identified. A high degree of antifungal resistance was observed. Fusarium isolates exhibited lower MICs to luliconazole and terbinafine compared with amphotericin B and voriconazole, which in turn were significantly more active than amorolfine, fluconazole and itraconazole. MALDI-ToF MS showed good performance in Fusarium species with an adapted Bruker library and expanded database. Fusarium isolates exhibited lower MICs to luliconazole and terbinafine compared to amphotericin B and voriconazole.


Perceptions of Transgender and Nonbinary Persons Toward Breast and Cervical Cancer Development, Screening, and Potential Impact on Gender-Affirming Hormone Therapy.
Approximately 1.6% of adult Americans identify as transgender (TG) or nonbinary (NB) and many take gender-affirming hormone therapy (GAHT). Little data exist to inform breast and cervical cancer risks, gender-specific screening guidelines, and inclusive cancer treatment algorithms that consider GAHT. We aimed to assess TGNB persons' perceptions on breast and cervical cancer development, screening knowledge and practices, and attitude toward GAHT in the setting of a hormone receptor-positive breast cancer diagnosis. This single-institution survey study was conducted through an LGBTQ+ focused clinic from 2021 to 2022. Noncisgender patients age ≥ 18 years who were English speaking were eligible to participate. A 5-point Likert scale was used to assess concern of developing breast (all participants) and cervical cancer (those assigned female sex at birth). Demographic and quantitative variables were examined in comparison with responses via chi-squared tests. Eighty-six participants completed the survey: 43% TG men, 24% TG women, and 20% NB persons. Most (84.9%) were age < 40 years, and 86% were non-Hispanic White. The majority were unaware of breast (77%) or cervical (60%) cancer screening recommendations for their sex assigned at birth or their gender. Approximately 35% reported concern regarding breast cancer development and of those age > 40 years; only 50% had undergone screening mammography. Of those assigned female sex at birth with an intact cervix, 47% were concerned about cervical cancer development and 46.6% had a Papanicolaou smear within the past 5 years. Nearly all (87.2%) were on GAHT, and 35.1% reported they would not consider stopping GAHT if diagnosed with a hormone receptor-positive breast cancer. The findings support the need for patient and provider education on screening options and large prospective cohort data to elucidate optimal gender-specific screening guidelines and treatment algorithms. [Media: see text].


Determination of oversulfated chondroitin sulfate and dermatan sulfate impurities in heparin by capillary electrophoresis.
Recently, oversulfated chondroitin sulfate (OSCS) present in certain lots of heparin was identified as the toxic contaminant responsible for severe side effects following intravenous heparin administration. The United States Pharmacopeia (USP) and European Pharmacopeia (Eur.Ph.) announced an immediate revision of their monographs for heparin sodium by adding two US Food and Drugs Administration-recommended tests for OSCS based on nuclear magnetic resonance and capillary electrophoresis (CE). However, the proposed CE method provides only partial separation of the OSCS contaminant from heparin, thereby hindering appropriate impurity profiling. Here we present an improved CE method that is especially useful for the reliable quantification of OSCS in heparin samples, but also allows determination of the common impurity dermatan sulfate (DS). Parameters such as type and concentration of background electrolyte, capillary temperature, sample concentration and injection volume were investigated and optimized. Enhancement of the OSCS-heparin separation was achieved by using high concentrations of Tris phosphate (pH 3.0) as background electrolyte. High currents and excessive Joule heating were prevented by employing fused-silica capillaries with an internal diameter of 25 microm. Good separations of OSCS, heparin and DS are obtained within 17 min. The method permits injection of relatively high heparin concentrations (up to 50 mg/ml) and large sample volumes (up to 5% of the capillary volume) allowing OSCS and DS determination in heparin down to the 0.05% and 0.5% (w/w) level, respectively. The CE method is shown to be repeatable and linear (R(2)>0.99) for OSCS, heparin and DS. CE analyses of OSCS-contaminated heparin samples and different heparin standards further demonstrate the utility of the method.


The use of atypical antipsychotics and the risk of breast cancer.
To determine whether atypical antipsychotics, when compared to typical antipsychotics, increase the risk of breast cancer. We conducted a retrospective cohort study using a nested case-control analysis within the United Kingdom General Practice Research Database population. We identified all female patients prescribed at least one antipsychotic (either typical or atypical), between 1 January 1988 and 31 December 2007, with follow-up until 31 December 2010. All incident cases of breast cancer were identified and matched up to 10 controls. Adjusted rate ratios (RR) of breast cancer associated with ever use of atypical antipsychotics was compared to ever use of typical antipsychotics. The cohort included 106,362 patients prescribed antipsychotics during the study period. During a mean follow-up of 5.3 years, 1237 patients were diagnosed with breast cancer (overall rate: 2.7 per 1000/year). Compared to patients who only used typical antipsychotics, exclusive users of atypical antipsychotics were not an increased risk of breast cancer (RR: 0.81, 95% CI: 0.63, 1.05). These results remained consistent after considering specific atypical antipsychotics known to significantly increase prolactin levels such as risperidone (RR: 0.86, 95% CI: 0.60, 1.25). Furthermore, no dose-response was observed in terms of cumulative duration of use and cumulative dose in olanzapine equivalents. The results of this study should provide reassurance that compared to typical antipsychotics, atypical antipsychotics do not increase the risk of breast cancer.


From the child to the young adult: sex differences in the antecedents of psychological problems. A retrospective study over ten years.
Three hundred children chosen at random from the general population were examined at 9 years of age and seen again at 20. The results of the study emphasize the discontinuity of psychological problems between childhood and adulthood: serious difficulties in psychological functioning at age 9 do not predict disturbances at 20, but psychological disturbances at 20 may originate in adolescence. The overrepresentation, at age 9, of boys at risk disappears at 20, at which point there is a slightly higher proportion of women amongst the "cases". The factors which influence the mental health of girls and boys are analysed and discussed.


TDP-43 proteinopathy occurs independently of autophagic substrate accumulation and underlies nuclear defects in Niemann-Pick C disease.
Neuronal cytoplasmic inclusions of TAR-DNA binding protein of 43 kDa (TDP-43) are a pathological hallmark of diverse neurodegenerative disorders, yet the processes that mediate their formation and their functional significance remain incompletely understood. Both dysfunction in autophagy and neuroinflammation have been linked to TDP-43 mislocalisation. Here, we investigate TDP-43 proteinopathy in Niemann-Pick type C disease (NPC), an autosomal recessive lysosomal storage disease (LSD) distinguished by the accumulation of unesterified cholesterol within late endosomes and lysosomes. NPC is characterised by neurodegeneration, neuroinflammation and multifocal disruption of the autophagy pathway. We utilised immunohistochemistry, confocal microscopy, electron microscopy and biochemical and gene expression studies to characterise TDP-43 pathology and autophagic substrate accumulation in Npc1-deficient mice. In the NPC brain, cytoplasmic TDP-43 mislocalisation was independent of autophagic substrate accumulation. These pathologies occurred in distinct neuronal subtypes, as brainstem cholinergic neurons were more susceptible to TDP-43 mislocalisation, whereas glutamatergic neurons exhibited hallmarks of autophagic dysfunction. Furthermore, TDP-43 mislocalisation did not co-localise with markers of stress granules or progress to ubiquitinated aggregates over months in vivo, indicating a stable, early stage in the aggregation process. Neither microgliosis nor neuroinflammation were sufficient to drive TDP-43 proteinopathy in the NPC brain. Notably, cytoplasmic TDP-43 co-localised with the nuclear import factor importin α, and TDP-43 mislocalised neurons demonstrated nuclear membrane abnormalities and disruption of nucleocytoplasmic transport. Our findings highlight the relationship between LSDs and TDP-43 proteinopathy, define its functional importance in NPC by triggering nuclear dysfunction, and expand the spectrum of TDP-43 pathology in the diseased brain.


Postprandial systolic blood pressure responses of older people in residential care: association with risk of falling.
The association between postprandial blood pressure, falls and medications is controversial. To investigate cardiovascular responses to meals in elderly people together with clinical associations and therapeutic issues. A cross-sectional observational study of 179 semi-independent older people (age 83.2 +/- 7.0 years) in residential care facilities was undertaken. Data on the frequency of falls, medical and medication history and measurement of blood pressure before and after a breakfast meal, and then after standing and walking after the meal were documented. Postprandial hypotension (>/=20 mm Hg fall in systolic blood pressure (SBP)) and low absolute SBP (</=115 mm Hg) were common after meals and exacerbated by standing. Risk factors for low postprandial SBP included use of selective serotonin reuptake inhibitors (OR = 4.3, CI 1.4-13.1, p = 0.006), antipsychotic medications (OR = 5.2, CI 1.4-19.2, p = 0.007) and a history of smoking (OR = 4.7, CI 1.5-14.9, p = 0.005). Antihypertensive therapy ameliorated the postprandial decline in blood pressure. The incidence of falls was increased only in the 10% of subjects with low postprandial SBP. Significant adverse cardiovascular change is common after meals in older people and a postprandial SBP of <115 mm Hg was associated with a history of falls. Selective serotonin reuptake inhibitors and antipsychotic medications were associated with low postprandial SBP, which may provide a mechanism for their association with falls. Hypertension was positively and antihypertensive therapy negatively associated with postprandial hypotension.


Ultraviolet action spectra for photobiological effects in cultured human lens epithelial cells.
The action spectrum for cell killing by UV radiation in human lens epithelial (HLE) cells is not known. Here we report the action spectrum in the 297-365 nm region in cultured HLE cells with an extended lifespan (HLE B-3 cells) and define their usefulness as a model system for photobiological studies. Cells were irradiated with monochromatic radiation at 297, 302, 313, 325, 334 and 365 nm. Cell survival was determined using a clonogenic assay. Analysis of survival curves showed that radiation at 297 nm was six times more effective in cell killing than 302 nm radiation; 297 nm radiation was more than 260, 590, 1400 and 3000 times as effective in cell killing as 313, 325, 334 and 365 nm radiation, respectively. The action spectrum was similar in shape to that for other human epithelial cell lines and rabbit lens epithelial cells. The effect of UV radiation on crystallin synthesis was also determined at different wavelengths. To determine whether exposure to UV radiation affects the synthesis of beta-crystallin, cells were exposed to sublethal fluences of UV radiation at 302 and 313 nm, labeled with [35S]methionine and the newly synthesized beta-crystallin was analyzed by immunoprecipitation and western blotting using an antibody to beta-crystallin. The results show a decrease in crystallin synthesis in HLE cells irradiated at 302 and 313 nm at fluences causing low cytotoxicity. The effect of radiation on membrane perturbation was determined by measuring enhancement of synthesis of prostaglandin E2 (PGE2). Synthesis of PGE2 occurs at all UV wavelengths tested in the 297-365 nm region.(ABSTRACT TRUNCATED AT 250 WORDS)


Albumin used in human IVF contain different levels of lipids and modify embryo and fetal growth in a mouse model.
Different commercial human embryo culture mediums can alter embryo quality and change birthweight. One component that could be contributing to variations but is not widely investigated is human serum albumin (HSA). HSA plays a multitude of roles during embryo culture and is a carrier for molecules including lipids. It remains unclear if lipid composition of HSA varies among commercial products and its effects on embryo quality, implantation, and fetal outcomes are relatively unknown. Utilizing a mouse model of embryo culture, we cultured zygotes until the blastocyst stage (72-h culture) in G1/G2 containing either Vitrolife HSA, Sage HSA, or Recombinant HSA at 10%. Blastocyst quality (development, total cell number, superoxide generation), blastocyst lipid content (neutral lipids, non-esterified fatty acids, phospholipids, and triglycerides), implantation, and fetal lengths and weights were assessed. Fatty acid quantification of HSA source was assessed by standard thin-layer chromatography. Sage HSA had the greatest fatty acid composition, with an eightfold increase in saturated fatty acids. This coincided with reduced blastocyst development, increased superoxide generation, neutral lipids and triglycerides levels of blastocysts, and decreased implantation rates (p < 0.05). Unexpectedly, while Recombinant HSA had the lowest overall lipids it had 70-fold increase in palmitoleic acid and the lowest fetal weights (p < 0.05). Indicates the importance of a balance between different types/amount of lipids, and an "optimal ratio" required for embryo and fetal development. Therefore, the lipid content of HSA should be considered when choosing a suitable HSA source for use in clinical IVF.


Modification of DNA by bile acids: a possible factor in the etiology of colon cancer.
Bile acids have been implicated as promoters and cocarcinogens in the etiology of colon cancer and as comutagens and mutagens in bacteria. These observations suggest the hypothesis that bile acids may interact directly with DNA. We treated the single stranded circular DNA of phage M13 with bile acids and found that the transfection efficiency of this DNA declined up to a 1000-fold. This result suggests that bile acids can damage DNA and thus may play an important role in the etiology of colon cancer.


Special Issue: Viral Infections in Companion Animals.
Companion animals, such as cats, dogs, horses and exotic species, play an important role in society; more than 600 million cats and 900 million dogs live closely with humans worldwide [...].


Intracellular Na+ overload causes oxidation of CaMKII and leads to Ca2+ mishandling in isolated ventricular myocytes.
An increase of late Na(+) current (INaL) in cardiac myocytes can raise the cytosolic Na(+) concentration and is associated with activation of Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) and alterations of mitochondrial metabolism and Ca(2+) handling by sarcoplasmic reticulum (SR). We tested the hypothesis that augmentation of INaL can increase mitochondrial reactive oxygen species (ROS) production and oxidation of CaMKII, resulting in spontaneous SR Ca(2+) release and increased diastolic Ca(2+) in myocytes. Increases of INaL and/or of the cytosolic Na(+) concentration led to mitochondrial ROS production and oxidation of CaMKII to cause dysregulation of Ca(2+) handling in rabbit cardiac myocytes.


Preservation of the inferior mesenteric artery in laparoscopic nerve-sparing colorectal surgery for endometriosis.
Laparoscopic rectosigmoid resection for endometriosis is usually performed with the section of the inferior mesenteric artery (IMA) distal to the left colic artery (low-tie ligation). This study was to determine outcomes in IMA-sparing surgery in endometriosis cases. A single-center retrospective study based on the analysis of clinical notes of women who underwent laparoscopic rectosigmoid segmental resection and IMA-sparing surgery for deep infiltrating endometriosis with bowel involvement between March the 1st, 2018 and February the 29th, 2020 in a referral hospital. During the study period, 1497 patients had major gynecological surgery in our referral center, of whom 253 (17%) for endometriosis. Of the 100 patients (39%) who had bowel endometriosis, 56 underwent laparoscopic nerve-sparing rectosigmoid segmental resection and IMA-sparing surgery was performed in 53 cases (95%). Short-term complications occurred in 4 cases (7%) without any case of anastomotic leak. Preservation of the IMA in colorectal surgery for endometriosis is feasible, safe and enables a tension-free anastomosis without an increase of postoperative complication rates.


The influence of porous media heterogeneity on smouldering remediation.
Self-sustained Treatment for Active Remediation (STAR) is a thermal remediation technology that uses smouldering, a flameless form of combustion, for destroying organic contaminants in soil. Injected cold air flowing through the soil to the treatment zone supports the release of sufficient energy to maintain a self-sustained reaction and the propagation of the reaction through the contaminated zone as long as the airflow local to the reaction exceeds a minimum value. However, the distribution and magnitude of air flux vectors can be complex in the heterogeneous environment common at contaminated sites. This research presents the first investigation of smouldering remediation under varying degrees and patterns of permeability heterogeneity. Nine experiments examined smouldering remediation in contaminated layers of varying permeability arranged alone and in contrasting layers in series, in parallel, and in two distinct complex patterns. The results suggest that smouldering can successfully propagate across layer boundaries and through layers in series regardless of their permeability (at least down to 1 x 10-12 m2). However, fine layers were not smouldered for layers in parallel with a permeability ratio ≥ 3:1. Numerical modelling of these cases with a published smouldering model revealed that this occurred due to insufficient airflow in the fine layers in some cases, or conductive heat transfer (thermal coupling) between parallel layers in other cases. The more complex heterogeneity patterns underscored the importance of the connected length of the higher permeability pathway on airflow distribution and therefore on smouldering propagation. Disconnected coarse zones supported smouldering in both coarse and fine zones while connected coarse zones kept smouldering in the coarse pathway while bypassing fine zones. Overall, this research provides unique insights into understanding heterogeneous scenarios to ensure the successful application of smouldering remediation.


Effects of dicopper oxide and copper sulfate on growth performance and gut microbiota in broilers.
An experiment was conducted to determine the effects of two sources of copper (Cu) from copper sulfate (CuSO4) and dicopper oxide (Cu2O, CoRouge) at three levels of inclusion (15, 75, and 150 mg/kg) on growth performance and gut microbiota of broilers. A total of 840 one-d-old male chickens (Ross 308) were weighed and randomly allocated to seven dietary treatments: negative control (NC, a basal diet without Cu addition), and the NC supplemented with 15, 75, or 150 mg Cu/kg from CuSO4 or Cu2O (12 replicate pens/treatment, 10 chicks per pen). Broilers were challenged by reusing an old litter with high concentrations in Clostridium perfringens to promote necrotic enteritis. Broiler performance was registered at d 21, 35, and 42. Excreta samples were collected at d 14, 28, and 42 for antimicrobial resistance (AMR) analyses. At d 43, one broiler per pen was euthanized to obtain ileal content for microbial characterization. Body weight d 35 and daily gain d 42 improved (P < 0.05) in Cu2O as Cu dose inclusion increased from 15 mg/kg to 150 mg/kg. Supplementation of 150 mg/kg of Cu from Cu2O decreased the abundance (P < 0.01) of some families such as Streptococcaceae and Corynebacteriaceae and increased the abundance (P < 0.05) of some commensal bacteria like Clostridiaceae and Peptostreptococcaceae. Phenotypic AMR was not different among treatments on d 14 and 28. Isolated Enterococcus spp. from broilers fed the NC diet on d 42 showed higher (P < 0.05) resistance to enrofloxacin, gentamicin, and chloramphenicol compared with Cu treatments. By contrast, the isolated Escherichia coli from broilers fed 150 mg/kg of Cu, either from CuSO4 or Cu2O, showed higher (P < 0.05) resistance to streptomycin and chloramphenicol compared to the NC. This study suggests that supplementing 150 mg/kg of Cu from Cu2O establishes changes in the gut microbiota by regulating the bacterial population in the ileum, which may explain the positive impact on broilers' growth performance.


Helicobacter pylori infection status in relation to antibiotic and non-steroidal prescribing in patients on maintenance treatment for chronic duodenal ulcer.
It has been suggested that establishing Helicobacter pylori infection status is irrelevant prior to H. pylori eradication treatment in chronic duodenal ulcer, as virtually all may benefit from therapy. The aim of the present study was (i) to determine the prevalence of active H. pylori infection in patients with proven chronic duodenal ulcer on long-term H2-antagonist prophylactic treatment and whether knowledge of this would influence the use of eradication therapy and (ii) to assess other factors which might influence the clinical diagnosis or H. pylori status, such as non-steroidal and antibiotic use. One hundred and forty-five patients receiving long-term H2-antagonists for chronic duodenal ulcer were recruited. Their case records and a prescribing database were reviewed. Patients underwent endoscopy with biopsies for rapid urease test, histology and H. pylori culture. Serum was immunoblotted and an enzyme-linked immunosorbent assay for H. pylori was performed. Of the 145 patients, 128 (88%) were H. pylori biopsy positive. Twelve of the 17 H. pylori biopsy-negative patients had anti-H. pylori immunoglobulin G (IgG) antibodies and 10 of the 17 H. pylori-negative patients had previously received antibiotics for other indications. Nine patients were exposed to non-aspirin non-steroidal anti-inflammatory drugs (NSAIDs) and one had additional aspirin exposure. Only 11.7% of patients on maintenance treatment for chronic duodenal ulcer had no current infection with H. pylori, although more than 70% of these had serological evidence of previous infection. Confirmation of active infection may be indicated where there is a history of NSAID or antibiotic exposure and may result in more precise targeting of eradication therapy, thus avoiding unnecessary and potentially hazardous treatment.


Production of glutathione using a bifunctional enzyme encoded by gshF from Streptococcus thermophilus expressed in Escherichia coli.
Glutathione (GSH) is one of the most ubiquitous non-protein thiols that is involved in numerous cellular activities. The gene coding for a novel bifunctional enzyme catalyzing the reaction for glutathione synthesis, gshF, was cloned from Streptococcus thermophilus SIIM B218 and expressed in Escherichia coli JM109. In the presence of the precursor amino acids and ATP, the induced cells of E. coli JM109 (pTrc99A-gshF) could accumulate 10.3 mM GSH in 5 h. The S. thermophilus GshF was insensitive to feedback inhibition caused by GSH even at 20 mM. At elevated concentrations of the precursor amino acids and ATP, E. coli JM109 (pTrc99A-gshF) produced 36 mM GSH with a molar yield of 0.9 mol/mol based on added cysteine and of 0.45 mol/mol based on added ATP. When ATP was replaced with glucose, E. coli JM109 (pTrc99A-gshF) produced 7 mM in 3h. Saccharomyces cerevisiae was used to generate ATP for GSH production. In the presence of glucose and the pmr1 mutant of S. cerevisiae BY4742, JM109 (pTrc99A-gshF) produced 33.9 mM GSH in 12h with a yield of 0.85 mol/mol based on added l-cysteine. It is shown that the S. thermophilus GshF can be successfully used for GSH production.


Imaging of Cortical Microtubules in Plants Under Salt Stress.
The microtubule (MT) network is a highly dynamic subcellular structure playing an important role in the growth and development of plants, and it is able to respond to biotic and abiotic environmental signals. Recent literature shows that microtubules play a key role in the tolerance of plants to salt stress. For example, salt stress induces microtubules to undergo a process of depolymerization-repolymerization, which is necessary for Arabidopsis seedlings to survive under these conditions. However, the potential cellular and molecular mechanisms still need to be further studied. Here, we describe the protocol for salt treatment of Arabidopsis seedlings and imaging the MT array by confocal laser scanning microscopy. We also introduce the AnalyzeSkeleton (2D/3D) plugin for quantitative analysis of the microtubule array after salt stress. The application of such an image processing method can rapidly develop an appreciation of the role of microtubules in the salt stress response of plants.


Improving medium chain fatty acid production in Escherichia coli by multiple transporter engineering.
Medium-chain fatty acids (C6-C10, MCFAs) represent valuable molecules for food and pharmaceutical industries. However, MCFAs in Escherichia coli induce stresses and toxicity, reducing final yields. These phenotypes could be alleviated if MCFA export from cells was accelerated, yet information on proteins responsible for exporting MCFAs remains limited. Here, a panel of 17 genes or operons was screened. Overexpressing resistance nodulation cell division family transporter acrE, mdtE and mdtC increased MCFA titer by 46.4%, 65.2% and 33.8%, whereas overexpressing transcriptional activators soxS, marA and multidrug efflux pumps cmr, fadL decreased MCFA titer by 42.5%, 27.8%, 37.6% and 29.5%, respectively. Furthermore, the effect of multiple transporter engineering was investigated. Overexpressing acrE, mdtE and mdtC together with deleting cmr increased MCFA production by more than two-fold. This study demonstrated the efficient efflux pump specifically for MCFAs and this multiple transporter engineering strategy can be employed to synthesize other bio-products toxic to microorganisms.


A decision analysis model for reducing carbon emission from coal-fired power plants and its compensatory units.
The increasing carbon dioxide level in the earth's atmosphere and continuously changing climate creates a significant challenge to sustainability in the world. It is not easy to control pollution due to carbon dioxide emissions from coal-fired power plants into the atmosphere. However, carbon capture technology provides an advantage for capturing carbon from power plants. Various researchers suggested the non-linear optimization model with post-combustion carbon capture technology in coal-fired power plants to reduce carbon emission. However, in their research articles, most researchers did not include loss of power due to retrofitting carbon capture technology in power plants and carbon emission from the compensatory power plant. This paper proposes a linear optimization model that minimizes the emission release from the power plant and its compensatory plant by appropriate selection of carbon capture technology. Our proposed model incorporates loss of power due to adopting carbon capture technology and emission release from the power plant and compensatory power plant in the problem formulation. We have also generated the Pareto curve that determines the trade-off solutions between emission release and the overall electricity cost. The applicability of our model is illustrated through power sector data from two Indian states. The net reduction of emissions in the two states are 27.17 % and 26.29 %, achieved by a mixed integer linear programming approach in coal-fired power plants. The model developed is generic and provides a sustainable environment for the generation of electricity.


Development of bioengineered human larynx.
To date, only two human laryngeal allotransplants have been reported and, although they were successful, both patients required life-long immunosuppression. A bioengineered human larynx could represent a possible alternative to allotransplantation. Human larynxes were decellularized enzymatically to obtain acellular matrices. Histological and molecular analysis demonstrated that all cellular components and nuclear material were removed. SEM showed that decellularized matrices retained the hierarchical structures of the native larynx, and mechanical tests demonstrated that the decellularization did not significantly impaired the biomechanically properties of the obtained matrices. Immunohistochemical staining found residual angiogenic factors after decellularization, and CAM analysis demonstrated that acellular laryngeal scaffolds induce a strong in vivo angiogenic response. Using a decellularization method, we are now able to obtain, in a short and clinically useful time, natural bioengineered laryngeal scaffolds which could be use for partial or total implantation in humans.


Comparisons of calorie restriction and structured exercise on reductions in visceral and abdominal subcutaneous adipose tissue: a systematic review.
Exercise and low-calorie diets are common approaches taken to produce an energy deficit for weight loss in obesity. Changes in visceral and abdominal subcutaneous fat associated with weight loss are important questions but have not yet been concluded. We investigated the relationship between changes in visceral (VAT) and subcutaneous adipose tissue (SAT) areas obtained by abdominal imaging with the change in total body fat. The relevant databases were searched through January 2021 according to the PRISMA guidelines. Sixty-five studies were included. We found that the change in total body fat was associated with changes in both VAT and abdominal SAT areas, but the relationship between total body fat and the abdominal SAT area appeared stronger. Baseline values of VAT and abdominal SAT area were similar in the three treatment groups (calorie restriction, calorie restriction plus exercise, and exercise alone). The reduction in abdominal SAT area for a loss of 1 kg of total body fat was about 10 cm2, which was similar among all the treatments. The change in VAT area (-26.3 cm2) was a similar level as the change in abdominal SAT area (-31.5 cm2) in the exercise, whereas in the calorie restriction with and without exercise, the change in VAT area (-33.6 and -51.6 cm2, respectively) was approximately half of the reduction of SAT area (-65.1 and -87.2 cm2, respectively). Absolute changes in VAT and abdominal SAT areas might differ between interventions for the exercise and calorie restriction with and without exercise.


Primer design for multiplexed genotyping.
Single-nucleotide polymorphism (SNP) genotyping can be carried out by annealing an oligonucleotide primer directly adjacent to the polymorphism and carrying out a single base extension using a polymerase reaction with labeled dideoxynucleotide triphosphates. This can be multiplexed by attaching a unique tag at the 5'-end of each oligonucleotide primer and binding the corresponding antitag to a DNA microarray or microbead. After the polymerase reaction, the tag-antitag system can be used to demultiplex the experiment. However, such an assay requires careful primer and tag design to avoid any crossreactivity among the primers, tags, antitags, and template sequence. A procedure for designing the primers is described in this chapter.


Improving the decolorization for textile dyes of a metagenome-derived alkaline laccase by directed evolution.
To obtain better performing laccases for textile dyes decolorization, random mutagenesis of Lac591, a metagenome-derived alkaline laccase, was carried out. After three rounds of error-prone PCR and high-throughput screening by assaying enzymatic activity toward the phenolic substrate 2,6-dimethoxyphenol (2,6-DMP), a mutant (Lac3T93) with remarkably improved enzymatic activity was obtained. Sequence analysis revealed that four amino acid substitutions (N40S, V55A, F62L, and E316V) were accumulated in the Lac3T93. Compared to the wild-type enzyme, the specific activity of Lac3T93 toward 2,6-DMP was increased to 4.8-fold (61.22 U/mg), and its optimal temperature and pH were changed to 60°C and 8.0 from 55°C and 7.5 of the wild-type enzyme, respectively. Furthermore, the degradation ability of Lac3T93 for textile dyes was investigated, and the new variant represented improved decolorization percentage for four industrial dyes with complex phenyl structure (Basic Blue 3, Methylene Blue, Bromophenol Blue, and Crystal Violet) and higher decolorization efficiency for Indigo Carmine than that of the parent enzyme. Furthermore, the decolorization percentage of Lac3T93 for five dyes in the absence of hydroxybenzotrizole (HBT) is clearly higher than those of the wild-type enzyme with 1 mM HBT, and HBT can further improve its decolorization ability.


Case report-An unusual diagnosis of a gastric phytobezoar in a middle-aged woman.
A gastric phytobezoar is an uncommon finding. The most frequent symptoms are dyspepsia, abdominal pain, nausea, and vomiting (Gastroenterology Report, 7, 2019, 74). It is an important differential to remember in patients with risk factors for developing phytobezoars because phytobezoars can be managed conservatively in most cases.


Dataset for the estimation of a new body fat measurement method.
Due to the increasing prevalence of obesity and its negative consequences worldwide on public health, body composition analysis is a central pillar to assess the nutritional status. Scientists could use datasets to develop a new body fat measurement formula. Using bioelectrical impedance analysis, we analyzed the total body composition of 345 patients (234 men and 111 women) aged between 18 and 60 years residing in the metropolitan area of Barranquilla, Colombia. They have the potential for predictive formula analysis enhancing the cooperation among scientists. Due to the obesity pandemic, new datasets from other populations are needed to develop a body fat basic mathematical equation formula that could be used worldwide to determine the prevalence of overweight and obesity in a specific population group predisposed to develop metabolic syndrome or death, secondary to high cardiovascular risk.


Evaluation of the accuracy of diagnostic coding for influenza compared to laboratory results: the availability of test results before hospital discharge facilitates improved coding accuracy.
Assessing the accuracy of diagnostic coding is essential to ensure the validity and reliability of administrative coded data. The aim of the study was to evaluate the accuracy of assigned International Classification of Diseases version 10-Australian Modification (ICD-10-AM) codes for influenza by comparing with patients' results of their polymerase chain reaction (PCR)-based laboratory tests. A retrospective study was conducted across seven public hospitals in New South Wales, Australia. A total of 16,439 patients who were admitted and tested by either cartridge-based rapid PCR or batched multiplex PCR between January 2016 and December 2017 met the inclusion criteria. We calculated the sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of ICD-10-AM coding using laboratory results as a gold standard. Separate analyses were conducted to determine whether the availability of test results at the time of hospital discharge influenced diagnostic coding accuracy. Laboratory results revealed 2759 positive influenza cases, while ICD-10-AM coding identified 2527 patients. Overall, 13.7% (n = 378) of test positive patients were not assigned an ICD-10-AM code for influenza. A further 5.8% (n = 146) patients with negative test results were incorrectly assigned an ICD-10-AM code for influenza. The sensitivity, specificity, PPV and NPV of ICD-10-AM coding were 93.1%; 98.9%; 94.5% and 98.6% respectively when test results were received before discharge and 32.7%; 99.2%; 87.8% and 89.8% respectively when test results were not available at discharge. The sensitivity of ICD-10-AM coding varied significantly across hospitals. The use of rapid PCR or hospitalisation during the influenza season were associated with greater coding accuracy. Although ICD-10-AM coding for influenza demonstrated high accuracy when laboratory results were received before discharge, its sensitivity was substantially lower for patients whose test results were not available at discharge. The timely availability of laboratory test results during the episode of care could contribute to improved coding accuracy.


Part I: Vaccines for solid tumours.
Active specific immunotherapy holds great potential in the search for new therapeutic approaches for patients with cancer. Much preclinical and clinical evidence has shown that the immune system can be polarised against malignant cells by several vaccination strategies. Although no anticancer vaccine can be recommended outside clinical trials at present, tumour response and immunological findings in animals and humans should prompt researchers to investigate further the potential of this biotherapy. We summarise strategies for cancer vaccines so far implemented in the clinical setting, report the results of more than 200 clinical trials published over the past two decades, and discuss insights into preclinical tumour immunology that might aid the design of the next generation of cancer vaccines.


Sequential Versus Individual Saphenous Vein Grafting During Coronary Arterial Bypass Surgery.
Although multiarterial grafting or bilateral mammary artery use is being increasingly emphasized for contemporary coronary artery bypass grafting (CABG) practice, saphenous vein graft (SVG) is still the most frequently used CABG conduit, and it accounts for 80% of all CABG conduits. Research focusing on modifiable surgical factors such as anastomosis technique, however, is scarce. This study aimed to compare clinical outcomes and graft patency according to anastomosis methods of vein grafting. From January 2005 through December 2016, patients who underwent CABG using SVG either by a sequential or an individual grafting technique were enrolled in this study. Graft patency was evaluated with coronary computed tomographic angiography. Propensity-score matching was used to compare the clinical outcomes and graft patency of these 2 grafting techniques to reduce treatment selection bias. Overall 2515 eligible patients, 1,037 in the sequential SVG graft group (41.3%) and 1478 (58.8%) in the individual SVG graft group were enrolled. After propensity-score matching, 901 matched pairs of patients and 891 matched pairs of grafts were included in the final outcome analysis. There were no significant differences in unadjusted (P = .83) and adjusted overall mortality (hazard ratio [HR], 0.96; 95% confidence interval [CI], 0.79 to 1.17; P = .67). The composite outcome of death, nonfatal myocardial infarction, and repeat revascularization also did not significantly differ between the sequential SVG and the individual SVG groups for both before (P = .20) and after matching (HR, 0.91; 95% CI, 0.75 to 1.09; P = .30). The sequential grafts showed superior patency as compared with the individual grafts for both before (P = .015) and after adjustment (HR, 0.61; 95% CI, 0.45 to 0.82; P < .001). The sequential grafting technique of SVG showed fairly acceptable safety and efficacy with superior long-term graft patency than individual grafts. Sequential SVG grafts perhaps can be a reasonable option as a second graft in CABG in some clinical situations.


Re-contact demographics and clinical characteristics of diabetic patients treated for a hypoglycaemic episode in the pre-hospital environment: a rapid literature review.
Diabetes mellitus has been referred to as an 'epidemic' and the World Health Organization reported 422 million people with the disease in 2014. Hypoglycaemia is common among emergency presentations, yet understanding around the utilisation of emergency medical services (EMS) for this is incomplete.Ambulance service referral pathways for those suitable to be treated in the community have been developed as a means of managing this growing demand. However, there is limited evidence to suggest how they should be constructed or implemented.The aim of this review was to examine patients who re-contacted the health services following EMS non-transport for a hypoglycaemic episode and to determine if risk factors could be identified. Medline/PubMed and CINAHL online databases were searched for papers published between 1998 and 2018 relating to re-contacts following an interaction with EMS. The Cochrane Library online database was also searched, as well as manual searches from key journals. Relevant clinical manuals, guidelines and specific grey literature were also hand searched. After duplicates were removed, 260 articles were identified, with 41 selected for full review. These were then reduced by excluding those that did not provide any data on re-contact rates/demographics. The remaining papers were then assessed using the Critical Appraisal Skills Programme (CASP) appraisal tool and those identified as of low quality were removed. This produced 17 papers for final inclusion. The literature demonstrates that ambulance clinicians can appropriately treat hypoglycaemia in the community and identify those requiring further assessment at emergency departments. However, due to the very nature of diabetes, repeat episodes will and do occur, regardless of community or emergency department management, but these are rarely in the acute phase. Some groups are higher risk, but thorough holistic assessment is vital for identifying those suitable for community management.


Expression of genes encoding the collagen-binding heat shock protein (Hsp47) and type II collagen in developing zebrafish embryos.
Hsp47 is a heat-shock protein which interacts with newly synthesize procollagen chains in the endoplasmic reticulum (ER) of collagen-secreting cells and is thought to assist in procollagen triple helix assembly and subsequent transport to the cis-Golgi. This is supported by studies which have reported that genes encoding collagen and Hsp47 are subject to co-ordinate increases and decreases in expression in cultured cells. However, limited information is available regarding hsp47 expression in vivo, particularly during early embryonic development when a variety of collagen genes are expressed. Here we show that the zebrafish hsp47 gene is expressed in a dynamic spatiotemporal pattern in developing embryos. Strong expression of hsp47 mRNA is co-incident predominantly with expression of the type II collagen gene (col2a1) in a number of chondrogenic and non-chondrogenic tissues including the notochord, otic vesicle and developing fins. Notochordal expression of both genes is disrupted in floating head (flh) and no tail (ntl) embryos, which lack properly differentiated notochords. Surprisingly, no hsp47 mRNA is detectable in the strongly col2a1-expressing cells of the floor plate and hypochord, indicating that the two genes are not strictly co-regulated. Finally, Northern blot analysis revealed two alternative transcripts of col2a1 which are expressed in distinct temporal patterns. Appearance of the larger transcript occurs following somitogenesis, a time which coincides with the co-activation of hsp47 and col2a1 gene expression in tissues outside of the notochord.


Diversification of mineralocorticoid receptor genes in a subterranean rodent, the naked mole-rat.
Naked mole-rats (Heterocephalus glaber) inhabit subterranean burrows in savannas and are, thus, unable to access free water. To identify their mechanism of osmoregulation in xeric environments, we molecularly cloned and analyzed the nuclear receptor subfamily 3 group C member 2 (NR3C2) gene encoding the mineralocorticoid receptor (MR), required for hormone-dependent regulation of genes contributing to body fluid homeostasis. Most vertebrates harbor a single MR homolog. In contrast, we discovered that MR is duplicated in naked mole-rats. The amino acid sequence of naked mole-rat MR1 is 90% identical to its mouse ortholog, and MR1 is abundantly expressed in the kidney and the nervous system. MR2 encodes a truncated protein lacking DNA- and ligand-binding domains of MR1 and is expressed in diverse tissues. Although MR2 did not directly transactivate gene expression, it increased corticosteroid-dependent transcriptional activity of MR1. Our results suggest that MR2 might function as a novel regulator of MR1 activity to fine-tune MR signaling in naked mole-rats.


Cyclophosphamide-induced immunological tolerance: recovery leads to a memory state.
Tolerance to SRBC has been induced in adult mice by the aid of cyclophosphamide, and the duration of the tolerant state has been tested by a second injection of antigen at different times thereafter. As evidenced by 19S and 7S PFC 4 days after challenge, tolerance in this model is of relatively short duration and leads spontaneously to a memory state.


[Causes of death in patients with HIV infection in two Tunisian medical centers].
Antiretroviral tritherapy has contributed to a considerable reduction in HIV-related mortality. The causes of death are dominated by opportunistic infections in developing countries and by cardiovascular diseases and cancer in developed countries. To determine the causes and risk factors associated with death in HIV-infected patients in two Tunisian medical centers. cross-sectional study of HIV-infected patients over 15 years treated at Sousse and Monastir medical centers between 2000 and 2014. Death was considered related to HIV if its primary cause was AIDS-defining illness or if it was due to an opportunistic infection of unknown etiology with CD4 < 50 cells/mm3; it was considered unrelated to HIV if its primary cause wasn't an AIDS defining illness or if it was due to an unknown cause if no information was available. Two hundred thirteen patients, 130 men (61%) and 83 women (39%), average age 40 ± 11 years were enrolled in the study. Fifty four patients died, the mortality rate was 5.4/100 patients/year. Annual mortality rate decreased from 5.8% in 2000-2003 to 2.3% in 2012-2014. Survival was 72% at 5 years and 67% at 10 years. Death events were associated with HIV in 70.4% of cases. The leading causes of death were pneumocystis carinii pneumonia and cryptococcal meningitis in 6 cases (11%) each. Mortality risk factors were a personal history of opportunistic infections, duration of antiretroviral therapy < 12 months and smoking. Strengthening screening, early initiation of antiretroviral therapy and fight against tobacco are needed to reduce mortality in patients infected with HIV in Tunisia.


A comparison of glycoprotein biosynthesis in benign and malignant squamous epithelioma and normal epidermis.
Protein and N-linked glycoprotein biosynthesis was studied in histologically verified normal epidermis, actinic keratoses, keratoacanthoma, intra-epidermal carcinoma and squamous-cell carcinoma using polyacrylamide gel electrophoresis (PAGE). The PAGE profiles of 3[H]-leucine-labelled proteins and 3[H]-mannose-labelled glycoprotein from all disease states studied differed from each other and from normal epidermis. A large 3[H]-mannose-labelled glycoprotein region (band A) with a peak at 97-92 kDa appeared to indicate the presence of a relatively large proportion of basaloid cells in the tissue. An associated peak in the region of 78-74 kDa also appeared in normal epidermis and what appeared to be non-invasive lesions. The main region of change in all lesions corresponded to the 66-34-kDa region (bands B and C). The absence of a group of glycoproteins and proteins in the 62-58-kDa region appeared to be specific for invasive squamous-cell carcinoma. All tumours showed a peak at 38-34 kDa which was not present in normal epidermis. Actinic keratosis had a pattern similar to normal epidermis except that the peaks of band B tended towards the higher-molecular-weight end of the band than those in normal epidermis and peaks at 28-22 kDa were seen. The latter seemed to correspond to the presence of a high proportion of spinous cells in the tissue sample.


Phenolic composition and biological activities of Juniperus drupacea Labill. berries from Turkey.
The present study was designed to define the phenolic profile and the biological potential of berries methanol extract of Juniperus drupacea Labill. from Turkey. The total phenolic content (Folin-Ciocalteau assay) was 48.06±0.99mgGAE/g extract. The HPLC-DAD-ESI-MS analysis allowed the determination of the complete phenolic profile of J. drupacea berries. Phenolic acids represented more than 60% of the total phenolics, and tyrosol was the major one (1324±0.64μg/g extract); within the flavonoids amentoflavone was detected as the main constituent (927±0.35μg/g extract). The extract exhibited good antioxidant properties, as determined by different in vitro models: DPPH test (IC(50) 0.38±0.02mg/mL), reducing power (12.63±0.14ASE/mL), Fe(2+) chelating ability (IC(50) 2.26±0.06mg/mL), and TBA test (IC(50) 2.47±1.13μg/mL). Cytotoxicity against Artemia salina was highlighted (LC(50) 489.47±27.8μg/mL), and a significant decrease (p⩽0.05; p⩽0.01) in HepG2 cells viability was observed at the higher concentrations (5-10μg/mL). The extract displayed good antibacterial activity towards Gram-positive bacteria and in particular Staphylococcus aureus was the most susceptible strain (MIC 78.12μg/mL).


Measurement of the refractive index at cryogenic temperature of absorptive silver thin films used as reflectors in a Fabry-Perot cavity.
Data on the refractive index of silver thin films are scarce in the literature, and largely dependent on both the deposition method and thickness. We measure the refractive index of silver films at cryogenic temperature with a technique that takes advantage of the absorption of the films and the corresponding peculiar properties of Fabry-Perot cavities: a frequency shift between the reflection and transmission peaks, together with a modified cavity bandwidth. We demonstrate a decrease in the real value of the refractive index, together with a decrease in its imaginary value at 4 K.


A Tutorial of EDTA: Extensive De Novo TE Annotator.
Transposable elements (TEs) are important contributors to genome structure and evolution. With the growth of sequencing technologies, various computational pipelines and software programs have been developed to facilitate TE identification and annotation. These computational tools can be categorized into three types based on their underlying approach: homology-based, structural-based, and de novo methods. Each of these tools has advantages and disadvantages. In this chapter, we introduce EDTA (Extensive de novo TE Annotator), a new comprehensive pipeline composed of high-quality tools to identify and annotate all types of TEs. The development of EDTA is based on the benchmarking results of a collection of TE annotation methods. The selected programs are evaluated by their ability to identify true TEs as well as to exclude false candidates. Here, we present an overview of the EDTA pipeline and a detailed manual for its use. The source code of EDTA is available at https://github.com/oushujun/EDTA .


New Evidence of the Bidentate Binding Mode in 3-MBA Protected Gold Clusters: Analysis of Aqueous 13-18 kDa Gold-Thiolate Clusters by HPLC-ESI-MS Reveals Special Compositions Aun(3-MBA)p, (n = 48-67, p = 26-30).
Gold clusters protected by 3-MBA ligands (MBA = mercaptobenzoic acid, -SPhCO2H) have attracted recent interest due to their unusual structures and their advantageous ligand-exchange and bioconjugation properties. Azubel et al. first determined the core structure of an Au68-complex, which was estimated to have 32 ligands (3-MBA groups). To explain the exceptional structure-composition and reaction properties of this complex, and its larger homologs, Tero et al. proposed a "dynamic stabilization" via carboxyl O-H--Au interactions. Herein, we report the first results of an integrated liquid chromatography/mass spectrometer (LC/MS) analysis of unfractionated samples of gold/3-MBA clusters, spanning a narrow size range 13.4 to 18.1 kDa. Using high-throughput procedures adapted from bio-macromolecule analyses, we show that integrated capillary high performance liquid chromatography electrospray ionization mass spectrometer (HPLC-ESI-MS), based on aqueous-methanol mobile phases and ion-pairing reverse-phase chromatography, can separate several major components from the nanoclusters mixture that may be difficult to resolve by standard native gel electrophoresis due to their similar size and charge. For each component, one obtains a well-resolved mass spectrum, nearly free of adducts or signs of fragmentation. A consistent set of molecular mass determinations is calculated from detected charge-states tunable from 3- (or lower), to 2+ (or higher). One thus arrives at a series of new compositions (n, p) specific to the Au/3-MBA system. The smallest major component is assigned to the previously unknown (48, 26); the largest one is evidently (67, 30), vs. the anticipated (68, 32). Various explanations for this discrepancy are considered. A prospective is given for the various members of this novel series, along with a summary of the advantages and present limitations of the micro-scale integrated LC/MS approach in characterizing such metallic-core macro-molecules, and their derivatives.


Transcription factor MEF2D is required for the maintenance of MLL-rearranged acute myeloid leukemia.
Acute myeloid leukemia (AML) with MLL-rearrangement (MLL-r) comprises ∼10% of all AML cases and portends poor outcomes. Much remains uncovered on how MLL-r AML drives leukemia development while preventing cells from normal myeloid differentiation. Here, we identified that transcription factor MEF2D is a super-enhancer-associated, highly expressed gene in MLL-r AML. Knockout of MEF2D profoundly impaired leukemia growth, induced myeloid differentiation, and delayed oncogenic progression in vivo. Mechanistically, MEF2D loss led to robust activation of a CEBPE-centered myeloid differentiation program in AML cells. Chromatin profiling revealed that MEF2D binds to and suppresses the chromatin accessibility of CEBPE cis-regulatory regions. In human acute leukemia samples, MEF2D expression showed a strong negative correlation with the expression of CEBPE. Depletion of CEBPE partially rescued the cell growth defect and myeloid cell differentiation induced by the loss of MEF2D. Lastly, we show that MEF2D is positively regulated by HOXA9, and downregulation of MEF2D is an important mechanism for DOT1L inhibitor-induced antileukemia effects. Collectively, our findings suggest that MEF2D plays a critical role in human MLL-r AML and uncover the MEF2D-CEBPE axis as a crucial transcriptional mechanism regulating leukemia cell self-renewal and differentiation block.


Discontinuity in the competitive interaction of tumor cells with the tumor-bearing host--a hypotheses.
The hypothesis put forward here is that during the growth of a primary tumor ending in metastasis formation, phases occur in which 'unfavourable' conditions arise for the tumor cells. It is proposed that, in order to overcome these 'unfavourable' situations, the tumor cells subsequently secrete a substance designed to have a 'dysfunctional' effect on the normal cells. On the assumption of such a discontinuous secretion by the tumor cells, an investigation is made into the behaviour of certain tumors in humans, together with the findings of previously conducted tests. This leads to the formulation of possible concepts for prophylaxis and treatment. In conclusion, an attempt is made to locate what would be the precise target of such a hypothetical substance.


Examination of Corticospinal and Spinal Reflexive Excitability During the Course of Postoperative Rehabilitation After Anterior Cruciate Ligament Reconstruction.
To investigate corticospinal and spinal reflexive excitability and quadriceps strength in healthy athletes and athletes after anterior cruciate ligament reconstruction (ACLR) over the course of rehabilitation. Prospective cohort study. Eighteen athletes with ACLR and 18 healthy athletes, matched by sex, age, and activity, were tested at (1) 2 weeks after surgery, (2) the "quiet knee" time point, defined as full range of motion and minimal effusion, and (3) return to running, defined as achieving a quadriceps index of 80% or greater. We measured (1) corticospinal excitability, using resting motor threshold (RMT) and motor-evoked potential amplitude at a stimulator intensity of 120% of RMT (MEP120) to the vastus medialis, (2) spinal reflexive excitability, calculating the ratio of the maximal Hoffmann reflex to the maximal M-wave to the vastus medialis, and (3) isometric quadriceps strength. The ACLR group had higher RMTs in the nonsurgical limb and higher MEP120 in the surgical limb at all time points. The healthy-athlete group did not have interlimb differences. The RMT was positively associated with quadriceps strength 2 weeks after surgery; MEP120 was associated with quadriceps strength at all time points. Compared to healthy athletes, athletes after ACLR had altered corticospinal excitability that did not change from 2 weeks after surgery to the time of return to running. J Orthop Sports Phys Ther 2020;50(9):516-522. Epub 1 Aug 2020. doi:10.2519/jospt.2020.9329.


Anticancer and anti-inflammatory activities of 1,8-naphthyridine-3-carboxamide derivatives.
Several 1,8-naphthyridine-3-carboxamide derivatives (8-23) were synthesized and tested for in vitro cytotoxicity against eight cancer cell lines and a normal cell line. Compound 12 exhibited high cytotoxicity (IC(50)=1.37microM) in HBL-100 (breast) cell line while compounds 17 (IC(50)=3.7microM) and 22 (IC(50)=3.0microM) have shown high cytotoxicity in KB (oral) and SW-620 (colon) cell lines, respectively. The synthesized 1,8-naphthyridine-3-carboxamides were also evaluated for anti-inflammatory and myeloprotective activities, indicated by modulation in cytokine and chemokine levels secreted by dendritic cells.


A Genetic Evaluation System for New Zealand White Rabbit Germplasm Resources Based on SSR Markers.
At present, there is an abundance of quality domestic rabbit breeds in China. However, due to the lack of technical standards for the genetic evaluation of rabbit germplasm resources, there have been a number of problems, such as poor breed conservation. By studying the genetic diversity of 130 New Zealand white rabbits (regardless of generation), we obtained the best simple sequence repeat (SSR) marker combination. We found that, when using microsatellite markers for the effective genetic evaluation of domestic rabbits, the number of records should be greater than 60 and the marker number more than 22. Through the comparative analysis of 30 combinations of 22 markers, the optimal combination of 22 markers was determined, and the 22 SSR polymorphic loci were distributed on different chromosomes. We performed a genetic analysis of 200 New Zealand white rabbits corresponding to two generations, using the best SSR polymorphic loci combination. There were no significant differences in the genetic diversity parameters between the two generations of rabbits (p > 0.05), indicating that the characteristics of this excellent rabbit germplasm have been effectively preserved. At the same time, we verified that the established method can be used to evaluate the breed conservation of rabbit germplasm resources.


Pharmacokinetics of oral L-isoidide mononitrate in rats.
L-isoidide mononitrate (L-IIMN) is the most potent mononitrate vasodilator described so far in the literature. Since other mononitrates, such as isosorbide-5-mononitrate and isosorbide-2-mononitrate, have been shown not to be subject to first-pass metabolism, we examined the pharmacokinetics of L-IIMN after oral administration to determine whether this compound also exhibited this behavior. An oral dose of 2 mg kg-1 L-IIMN dissolved in normal saline was given to seven rats. Absorption of L-IIMN after dosing was rapid with an apparent absorption half-life of 9.5 +/- 3.6 min (mean +/- SD). Plasma L-IIMN concentrations peaked between 5 and 20 min after dosing and declined thereafter in an apparently monoexponential manner. The average elimination half-life was 11.9 +/- 1.7 min (mean +/- SD). Oral bioavailability was estimated to be about 50%. Thus, unlike the other mononitrates so far examined in the literature, L-IIMN exhibits incomplete bioavailability. This pharmacokinetic behavior, however, is consistent with its faster systemic clearance compared to other organic mononitrates.


The nuclear DICER-circular RNA complex drives the deregulation of the glioblastoma cell microRNAome.
The assortment of cellular microRNAs ("microRNAome") is a vital readout of cellular homeostasis, but the mechanisms that regulate the microRNAome are poorly understood. The microRNAome of glioblastoma is substantially down-regulated in comparison to the normal brain. Here, we find malfunction of the posttranscriptional maturation of the glioblastoma microRNAome and link it to aberrant nuclear localization of DICER, the major enzymatic complex responsible for microRNA maturation. Analysis of DICER's nuclear interactome reveals the presence of an RNA binding protein, RBM3, and of a circular RNA, circ2082, within the complex. Targeting of this complex by knockdown of circ2082 results in the restoration of cytosolic localization of DICER and widespread derepression of the microRNAome, leading to transcriptome-wide rearrangements that mitigate the tumorigenicity of glioblastoma cells in vitro and in vivo with correlation to favorable outcomes in patients with glioblastoma. These findings uncover the mechanistic foundation of microRNAome deregulation in malignant cells.


Risk factors for urinary retention after urogynecologic surgery: A retrospective cohort study and prediction model.
Postoperative urinary retention (POUR) is a common complication of urogynecological surgery. Our study aimed to identify demographic and perioperative risk factors to construct a prediction model for POUR in urogynecology. Our retrospective cohort study reviewed all patients undergoing pelvic reconstructive surgeries at our tertiary care center (Jan 1, 2013-May 1, 2019). Demographic, pre-, intra- and postoperative variables were collected from medical records. The primary outcome, POUR, was defined as (1) early POUR (E-POUR), failing initial trial of void or; (2) late POUR (L-POUR), requiring an indwelling catheter or intermittent catheterization on discharge. Risk factors were identified through univariate and multivariate logistic regression analyses. A clinical prediction model was constructed with the most significant and clinically relevant risk factors. In 501 women, 182 (36.3%) had E-POUR and 61 of these women (12.2% of the entire cohort) had L-POUR. Multivariate logistic regression revealed preoperative postvoid residual (PVR) over 200 ml (odds ratio [OR]: 3.17; p = 0.026), voiding dysfunction symptoms extracted from validated questionnaires (OR: 3.00; p = 0.030), and number of concomitant procedures (OR: 1.30 per procedure; p = 0.021) as significant predictors of E-POUR; preoperative PVR more than 200 ml (OR: 4.07; p = 0.011) and antiincontinence procedure with (OR: 3.34; p = 0.023) and without (OR: 2.64; p = 0.019) concomitant prolapse repair as significant predictors of L-POUR. A prediction model (area under the curve: 0.70) was developed for E-POUR. Elevated preoperative PVR is the most significant risk factor for POUR. Alongside other risk factors, our prediction model for POUR can be used for patient counseling and surgical planning in urogynecologic surgery.