Molecular modeling of LDLR aids interpretation of genomic variants.
Genetic variants in low-density lipoprotein receptor (LDLR) are known to cause familial hypercholesterolemia (FH), occurring in up to 1 in 200 people (Youngblom E. et al. 1993 and Nordestgaard BG et al. 34:3478-3490a, 2013) and leading to significant risk for heart disease. Clinical genomics testing using high-throughput sequencing is identifying novel genomic variants of uncertain significance (VUS) in individuals suspected of having FH, but for whom the causal link to the disease remains to be established (Nordestgaard BG et al. 34:3478-3490a, 2013). Unfortunately, experimental data about the atomic structure of the LDL binding domains of LDLR at extracellular pH does not exist. This leads to an inability to apply protein structure-based methods for assessing novel variants identified through genetic testing. Thus, the ambiguities in interpretation of LDLR variants are a barrier to achieving the expected clinical value for personalized genomics assays for management of FH. In this study, we integrated data from the literature and related cellular receptors to develop high-resolution models of full-length LDLR at extracellular conditions and use them to predict which VUS alter LDL binding. We believe that the functional effects of LDLR variants can be resolved using a combination of structural bioinformatics and functional assays, leading to a better correlation with clinical presentation. We have completed modeling of LDLR in two major physiologic conditions, generating detailed hypotheses for how each of the 1007 reported protein variants may affect function. KEY MESSAGES: • Hundreds of variants are observed in the LDLR, but most lack interpretation. • Molecular modeling is aided by biochemical knowledge. • We generated context-specific 3D protein models of LDLR. • Our models allowed mechanistic interpretation of many variants. • We interpreted both rare and common genomic variants in their physiologic context. • Effects of genomic variants are often context-specific.


Changes in diving behaviour and habitat use of provisioned whale sharks: implications for management.
Whale shark (Rhincodon typus) tourism is increasingly popular at predictable aggregations around the world, but only a few use provisioning to ensure close interactions. Understanding the effects of provisioning on the behaviour of this endangered species is critical to manage this growing industry. We recorded the diving behaviour and habitat use of juvenile whale sharks (n = 4) for a mean of 49.5 provisioned and 33.8 non-provisioned days using temperature-depth-recorders. We found that time spent at the surface (< 2 m) between 6 am and 1 pm increased ~ sixfold, while timing of deep dives shifted from 4-10 am to 10 am-2 pm, i.e. near or at the end of the provisioning activities. The shift might be related to a need to thermoregulate following a prolonged period of time in warmer water. These changes could have fitness implications for individuals frequently visiting the provisioning site. Based on recorded amount of time spent in warm waters and published Q10 values for ectotherms, we estimate a 7.2 ± 3.7% (range 1.3-17.8%) higher metabolic rate when sharks frequent the provisioning site. The observed behavioural, habitat use, and potential fitness shifts should be considered when developing guidelines for sustainable tourism, particularly in light of new provisioning sites developing elsewhere.


[Diagnostic informativeness of various psychopathologic symptom complexes].
A comparison of clinico-nosological and syndromal psychopathological diagnoses made in 81 patients as a result of follow-up examinations carried out in the periods of repeated forensic psychiatric examinations demonstrated a higher diagnostic reliability of some psychopathological symptom-complexes. It was organic and schizophrenic psychopathological symptom-complexes that were found to be of the greatest diagnostic informative value. These symptom-complexes enabled one to diagnose exactly the nosological nature of the psychic pathology as early, as in the initial period of the disease, or in atypical, clinically altered cases of its course.


Do individuals who develop transient low back pain exhibit different postural changes than non-pain developers during prolonged standing?
Previous literature has shown that individuals can develop transient low back pain (LBP) during a 2-h bout of unconstrained prolonged standing. The purpose of this study was to investigate the postural changes of individuals who develop LBP during standing (PD) and those who do not (NPD). Forty-one subjects (20 male, 21 female) participated in a 2-h prolonged standing occupational simulation and recorded their ratings of perceived LBP on a visual analog scale (VAS). Center of pressure changes (shifts, drifts, and fidgets) and body weight shifts were determined for each subject. All subjects showed an increase in the BW shift frequency and a decrease in average shift duration over the 2-h protocol. All NPD and female PD were consistent for many of the variables; however, male PD did not show similar patterns to the other groups, especially for anterior-posterior (AP) shift amplitude and total body weight asymmetry. Although gender differences between the pain groups were found, PD and NPD do not use different postural changes during unconstrained standing, showing that changes to postural control may be an "adaptive", rather than "causative" response to their LBP development. Future works should concentrate on bridging the literature seen in quiet standing before and after prolonged standing and the response of PD, as well as the effect of an exercise intervention on postural control, especially for male PD.


[Contribution of digestive system ultrasonography in the assessment of abdominal pain in children].
The quality of modern ultrasonography allows to study not only solid abdominal organs but also digestive walls and abdominal lymphnodes, leading to accurate diagnosis in many cases, and decreasing the use of more aggressive radiological procedures (upper GI baryum studies, baryum enema, intravenous pyelography, etc.). In dealing with abdominal pain in children, the plain XRay film of the abdomen and abdominal echography are performed immediately after history taking and physical examination, and are equally important in the work up of these cases.


Higher reproductive skew among birds than mammals in cooperatively breeding species.
While competition for limited breeding positions is a common feature of group life, species vary widely in the extent to which reproduction is shared among females ('reproductive skew'). In recent years, there has been considerable debate over the mechanisms that generate variation in reproductive skew, with most evidence suggesting that subordinates breed when dominants are unable to prevent them from doing so. Here, we suggest that viviparity reduces the ability of dominant females to control subordinate reproduction and that, as a result, dominant female birds are more able than their mammal counterparts to prevent subordinates from breeding. Empirical data support this assertion. This perspective may increase our understanding of how cooperative groups form and are stabilized in nature.


Downregulation of ACE2 expression by SARS-CoV-2 worsens the prognosis of KIRC and KIRP patients via metabolism and immunoregulation.
Background: Angiotensin-converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2) allow entry of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) into host cells and play essential roles in cancer therapy. However, the functions of ACE2 and TMPRSS2 in kidney cancer remain unclear, especially as kidneys are targets for SARS-CoV-2 infection. Methods: UCSC Xena project, the Cancer Genome Atlas (TCGA), and Gene Expression Omnibus (GEO) databases (GSE30589 and GSE59185) were searched for gene expression in human tissues, gene expression data, and clinical information. Several bioinformatics methods were utilized to analyze the correlation between ACE2 and TMPRSS2 with respect to the prognosis of kidney renal clear cell carcinoma (KIRC) and kidney renal papillary cell carcinoma (KIRP). Results: ACE2 expression was significantly upregulated in tumor tissue, while its downregulation was associated with low survival in KIRC and KIRP patients. TMPRSS2 was downregulated in KIRC and KIRP, and its expression was not correlated with patient survival. According to clinical risk factor-based prediction models, ACE2 exhibits predictive accuracy for kidney cancer prognosis and is correlated with metabolism and immune infiltration. In an animal model, ACE2 expression was remarkably downregulated in SARS-CoV-2-infected cells compared to in the control. Conclusion: ACE2 expression is highly correlated with various metabolic pathways and is involved in immune infiltration.it plays a crucial role than TMPRSS2 in diagnosing and prognosis of kidney cancer patients. The overlap in ACE2 expression between kidney cancer and SARS-CoV-2 infection suggests that patients with KIRC or KIRP are at high risk of developing serious symptoms.


Structural and functional characterization of the membrane-permeabilizing activity of Nicotiana occidentalis defensin NoD173 and protein engineering to enhance oncolysis.
Defensins are an extensive family of host defense peptides found ubiquitously across plant and animal species. In addition to protecting against infection by pathogenic microorganisms, some defensins are selectively cytotoxic toward tumor cells. As such, defensins have attracted interest as potential antimicrobial and anticancer therapeutics. The mechanism of defensin action against microbes and tumor cells appears to be conserved and involves the targeting and disruption of cellular membranes. This has been best defined for plant defensins, which upon binding specific phospholipids, such as phosphatidylinositol 4,5-bisphosphate (PIP2) and phosphatidic acid, form defensin-lipid oligomeric complexes that destabilize membranes, leading to cell lysis. In this study, to further define the anticancer and therapeutic properties of plant defensins, we have characterized a novel plant defensin, Nicotiana occidentalis defensin 173 (NoD173), from N. occidentalis. NoD173 at low micromolar concentrations selectively killed a panel of tumor cell lines over normal primary cells. To improve the anticancer activity of NoD173, we explored increasing cationicity by mutation, with NoD173 with the substitution of Q22 with lysine [NoD173(Q22K)], increasing the antitumor cell activity by 2-fold. NoD173 and the NoD173(Q22K) mutant exhibited only low levels of hemolytic activity, and both maintained activity against tumor cells in serum. The ability of NoD173 to inhibit solid tumor growth in vivo was tested in a mouse B16-F1 model, whereby injection of NoD173 into established subcutaneous tumors significantly inhibited tumor growth. Finally, we showed that NoD173 specifically targets PIP2 and determined by X-ray crystallography that a high-resolution structure of NoD173, which forms a conserved family-defining cysteine-stabilized-αβ motif with a dimeric lipid-binding conformation, configured into an arch-shaped oligomer of 4 dimers. These data provide insights into the mechanism of how defensins target membranes to kill tumor cells and provide proof of concept that defensins are able to inhibit tumor growth in vivo.-Lay, F. T., Ryan, G. F., Caria, S., Phan, T. K., Veneer, P. K., White, J. A., Kvansakul, M., Hulett M. D. Structural and functional characterization of the membrane-permeabilizing activity of Nicotiana occidentalis defensin NoD173 and protein engineering to enhance oncolysis.


Monoclonal antibodies to cowpox virus: polypeptide analysis of several major antigens.
Monoclonal antibodies directed against major antigens induced by cowpox virus (CPV) were produced. The specificities of these antibodies were established by immunoprecipitation, immunoblotting and several serological analyses, and from the cross-reactivities of these antibodies with cells infected with various other poxviruses, ectromelia virus (EV), vaccinia virus and Shope fibroma virus. The antibodies defined included ones reacting with each of the known major antigens of poxviruses, i.e. the common antigen of all poxviruses (probably NP antigen), the Orthopoxvirus-specific antigen (probably LS antigen), the haemagglutinin, the cell surface antigen, the common A-type inclusions in CPV and EV, and the antigen involved in neutralization.


Associations between common polymorphisms in TP53 and p21WAF1/Cip1 and phenotypic features of breast cancer.
The tumour suppressor gene TP53 and its downstream effector p21 are thought to play major roles in the development of breast cancer. We investigated three common sequence variants in TP53 and p21 for possible associations with the risk of breast cancer and with various phenotypic features of this disease. A total of 351 cases were available for study. Germline DNA obtained from female subjects of similar age but without cancer was used to estimate the TP53 and p21 genotype frequencies in a control population. A single nucleotide polymorphism in intron 2 of p21 was associated with slightly increased breast cancer risk (RR = 1.4, P = 0.011) and with well/moderately differentiated tumour histology (P = 0.029). The 16 bp insertion polymorphism in intron 3 of TP53 was associated with poor histological grade (OR = 2.3, P = 0.013) independently of other pathological features. The codon 31 polymorphism in p21 was strongly linked to negative progesterone receptor status (OR = 3.4, P = 0.0001), suggesting this variant may have functional significance for the progesterone signalling pathway in breast cancer. These results add to the growing body of evidence that genetic variants can influence not only the risk of breast cancer but also the disease phenotype.


Perioperative Bilateral Medial Medullary Infarction With "Snake Eyes Appearance": A Case Report.
Perioperative bilateral medial medullary infarction (BMMI) cases mimicking acute motor axonal neuropathy (AMAN) under general anesthesia have not been reported. We describe a patient who suffered flaccid quadriplegia and could not wean from mechanical ventilation after emergence from general anesthesia in cardiac surgery. A diagnosis of AMAN was considered, but intravenous immunoglobulin showed little efficacy. Magnetic resonance imaging of the patient later revealed BMMI with "snake eyes appearance," and he was found to have severe vertebral artery stenosis. Considering the association between severe coronary heart disease and cerebrovascular stenosis, we highlight the significance of preoperative evaluation and comprehensive management of the cerebrovascular system for certain patients.


Depth of phonological recoding in short-term memory.
In testing the hypothesis that surface phonetic form is included in short-term memory (STM) representation, the tone sandhi phenomenon in Mandarin Chinese was exploited, and, as a prerequisite, the hypothesis that tonal similarity affects STM of verbal material in a tone language was also tested. In Experiment 1, subjects recalled visually presented sequences of seven monosyllabic Chinese morphemes having either the same tone or different tones. More errors were made on the monotonal sequences than on the multitonal sequences, confirming the effect of tonal similarity on STM. In Experiment 2, subjects recalled visually presented sequences of disyllabic nonsense words. The sequences were designed in such a way that half of them were subject to the tone sandhi rule in Mandarin Chinese, whereas the other half were not. The consequence of applying the tone sandhi rule, as designed, was to make all the first characters in the sequences identical in pronunciation, thus creating potential phonological confusion. More errors, indeed, occurred on the sequences subject to the tone sandhi rule than on those not subject to it, indicating the existence of a surface phonetic representation in STM. The findings in this study provide further insight into the phonological mechanism of STM. Different accounts for this mechanism are also discussed in the light of the new findings.


Prediction of early (4-week) mortality in acute myeloid leukemia with intensive chemotherapy.
The progress with intensive chemotherapy and supportive care measures has improved survival in patients with newly diagnosed acute myeloid leukemia (AML). Given the recent development of effective low intensity therapies, an optimal decision on the therapy intensity may improve survival through the avoidance of early mortality. We reviewed the outcome of 3728 patients with newly diagnosed AML who received intensive chemotherapy between August 1980 and May 2020. Intensive chemotherapy was defined as a cumulative cytarabine dose ≥ 700 mg/m2 during induction therapy. We divided the whole cohort into a training and validation group at a 3:1 ratio. The population was divided into a training (2790 patients) and a validation cohort (938 patients). The median age was 55 years (range, 15-99). Among them, 442 patients (12%) had core-binding factor AML. Binary logistic regression identified older age, worse performance status, hyperbilirubinemia, elevated creatinine, hyperuricemia, cytogenetic abnormalities other than CBF and -Y, and pneumonia as adverse prognostic factors for an early 4-week mortality. This risk classification for early mortality was verified in the validation cohort of patients. In the validation cohort of more recently treated patients from 2000 to 2017, the 4-week mortality rates with intensive chemotherapy were 2%, 14%, and 50% in the low-, high-, and very high-risk group, respectively. The mortality rates with low intensity therapies were 3%, 9%, and 20%, respectively. The risk classification guides treatment intensity by the assessment of age, frailty, organ dysfunction, cytogenetic abnormality, and infection to avoid early mortality.


In vitro studies on the antibacterial activities of YM-13115, a new broad-spectrum cephalosporin.
The in vitro antibacterial activities of YM-13115, a new parenteral cephalosporin, were compared with those of ceftazidime, cefoperazone, and cefsulodin. The compound was highly active against the common members of the Enterobacteriaceae and 2 to 256 times more active than cefoperazone. YM-13115 was as active as ceftazidime against Citrobacter freundii, Proteus vulgaris, and Morganella morganii and two to four times more active than ceftazidime against Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, Enterobacter aerogenes, Serratia marcescens, Proteus mirabilis, Providencia rettgeri, and Providencia stuartii. The activity of YM-13115 against Pseudomonas aeruginosa (with MICs of 0.78 and 3.13 micrograms/ml for 50 and 90% of the isolates, respectively) was ca. 2 times that of ceftazidime, 4 times that of cefsulodin, and 16 times that of cefoperazone. Against Haemophilus influenzae YM-13115 was more active than ceftazidime. YM-13115 was less active than ceftazidime, cefoperazone, and cefsulodin against Staphylococcus aureus and Staphylococcus epidermidis. The concentrations of YM-13115 required to inhibit the growth of 90% of the isolates of Streptococcus pyogenes and Streptococcus pneumoniae were 0.78 and 1.56 microgram/ml, respectively, but concentrations above 100 micrograms/ml were required to inhibit Streptococcus faecalis. YM-13115 was not hydrolyzed by the common plasmid and chromosomal beta-lactamases. YM-13115 is extremely active against P. aeruginosa and members of the Enterobacteriaceae.


Age differences in the effects of domain knowledge on reading efficiency.
In the present study, the author investigated age differences in the effects of knowledge on the efficiency with which information is processed while reading. Individuals between 18 and 85 years of age, with varying levels of cooking knowledge, read and recalled a series of short passages within the domain of cooking. Reading efficiency was operationalized as time spent reading divided by the amount recalled for each passage. Results showed that reading efficiency increased with increasing levels of knowledge among older but not younger adults. Similarly, those with smaller working memory capacities showed increasing efficiency with increasing knowledge. These findings suggest that knowledge promotes a more efficient allocation policy that is particularly helpful in later life, perhaps due to age-related declines in working memory capacity.


Survival benefit of anti-angiogenic agents in patients with previously treated advanced gastric or gastroesophageal junction cancer: a meta-analysis.
There is a debate as to whether anti-angiogenic molecular agents can produce survival benefits in patients with previously treated advanced gastric cancer (GC) or gastroesophageal junction cancer (GEJC). We performed this meta-analysis of randomized trials to evaluate the survival outcomes of an anti-angiogenic agent versus placebo in the salvage treatment of advanced GC or GEJC. Electronic databases were searched for eligible studies. From the four studies, 910 patients with previously treated advanced GC or GEJC were included in the meta-analysis. Compared with placebo, anti-angiogenic targeted agents significantly improved progression-free survival (hazard ratio = 0.37 [95% confidence interval, 0.26-0.53], P < 0.00001). In terms of overall survival, anti-angiogenic agents induced 36% reduction in the risk for death (hazard ratio = 0.64 [95% confidence interval, 0.48-0.86], P = 0.002). In conclusion, this meta-analysis demonstrates that anti-angiogenic agents can prolong survival in patients with previously treated advanced GC or GEJC. This finding suggests that anti-angiogenic therapy can be a considerable option in patients who are not candidates for further chemotherapy.


Pseudomonas aeruginosa infection and inflammation during contact lens wear: a review.
Infection and inflammation during contact lens wear is often associated with microbial contamination of lenses. Several different types of microbes that colonize lenses can lead to infection and inflammation, but the most common cause of infection (microbial keratitis; MK) remains the Gram-negative bacterium Pseudomonas aeruginosa. P. aeruginosa has a battery of cell-associated and extracellular virulence factors it can use to initiate and maintain infection. Its ability to produce proteases, to either invade or kill corneal cells, and to coordinate expression of virulence factors via quorum-sensing have been shown to be important during MK. Another important factor that contributes to the destruction of the cornea during MK is excessive activation of the host defense system. P. aeruginosa can activate several pathways of the immune system during MK, and activation often involves receptors on the corneal epithelial cells called toll-like receptors (TLRs). These TLRs recognize e.g., lipopolysaccharide or flagella from P. aeruginosa and activate the epithelial cells to produce inflammatory mediators such as cytokines and chemokines. These cytokines or chemokines recruit white blood cells, predominantly polymorphonuclear leukocytes, to the infection in order that they can phagocytose and kill the P. aeruginosa. However, continued recruitment and presence of these polymorphonuclear neutrophils and other white blood cells in the corneal tissue leads to destruction of corneal cells and tissue components. This can ultimately lead to scarring and vision loss.


Downregulation of JMJD2a and LSD1 is involved in CK2 inhibition-mediated cellular senescence through the p53-SUV39h1 pathway.
Lysine methylation is one of the most important histone modifications that modulate chromatin structure. In the present study, the roles of the histone lysine demethylases JMJD2a and LSD1 in CK2 downregulation-mediated senescence were investigated. The ectopic expression of JMJD2a and LSD1 suppressed the induction of senescence-associated β-galactosidase activity and heterochromatin foci formation as well as the reduction of colony-forming and cell migration ability mediated by CK2 knockdown. CK2 downregulation inhibited JMJD2a and LSD1 expression by activating the mammalian target of rapamycin (mTOR)-ribosomal p70 S6 kinase (p70S6K) pathway. In addition, the downregulation of JMJD2a and LSD1 was involved in activating the p53-p21Cip1/WAF1-SUV39h1-trimethylation of the histone H3 Lys9 (H3K9me3) pathway in CK2-downregulated cells. Further, CK2 downregulation-mediated JMJD2a and LSD1 reduction was found to stimulate the dimethylation of Lys370 on p53 (p53K370me2) and nuclear import of SUV39h1. Therefore, this study indicated that CK2 downregulation reduces JMJD2a and LSD1 expression by activating mTOR, resulting in H3K9me3 induction by increasing the p53K370me2-dependent nuclear import of SUV39h1. These results suggest that CK2 is a potential therapeutic target for age-related diseases. [BMB Reports 2022;55(2): 92-97].


[99mTc-2-methoxy-isobutyl-isonitrile in the diagnosis of coronary heart disease: results of a multicenter study].
A total of 226 patients was studied in a phase-III multicenter trial to compare the results of stress/rest 99mTc-MIBI with 201TI scintigraphy and/or coronary arteriography. The evaluation of planar and SPECT imaging was based on a semiquantitative visual scoring. A congruent diagnosis was generated from all planar 201TI and 99mTc-MIBI scintigrams performed in 36 patients. In 62 of 63 patients, SPECT-studies with 99mTc-MIBI and 201TI led to an identical diagnosis. In one patient a positive result was obtained with 201TI-SPECT whereas 99mTc-MIBI was negative. Segmental agreement for 1509 SPECT segments was 85.7%, for 554 planar segments it was 86.6%. In comparison to coronary arteriography, performed in 180 patients, the overall sensitivity for the correct diagnosis of coronary artery disease (CAD) was 86% for planar MIBI scintigraphy or 92% for MIBI-SPECT. Overall specificity was 100% for planar imaging and 57% for SPECT. The low specificity of MIBI-SPECT was probably due to high prevalence of CAD in this study population. In a subgroup of 43 patients, who underwent coronary angiography, identical results were found with 99mTc-MIBI and 201TI. Only one patient showed a positive scintigraphic result with both 99mTc-MIBI and 201TI without angiographically proven stenosis greater than 50% or prior myocardial infarction. Vessel sensitivities in stenosed coronary arteries (greater than 50% stenosis) were 54% for the LAD and 87% for the RCX/RCA areas with MIBI-SPECT, or 51% or 79% for planar scintigraphy, respectively. Vessel specificities for SPECT were 65% in the LAD or 52% in the RCX/RCA, or 90% or 83% in planar imaging.(ABSTRACT TRUNCATED AT 250 WORDS)


The effects of caffeine ingestion on exercise-induced hypoalgesia: A pilot study.
Exercise acutely reduces pain sensitivity, termed exercise-induced hypoalgesia (EIH). The mechanisms underlying EIH remain unclear. Caffeine, a non-specific adenosine receptor antagonist has been shown to attenuate EIH in animals-suggesting the involvement of the adenosinergic system. This pilot study investigated the effects of caffeine on pain sensitivity following cycling exercise in college-aged men. Pressure pain threshold (PPT) and thermal pain threshold (TPT) were assessed in thirteen low caffeine consuming men prior to ingestion of a counter-balanced 5mg·kg(-1) dose of caffeine or a placebo (Pre), 60min following ingestion (Post-In), and then following a 15min bout of cycling exercise (Post-Ex) at an intensity eliciting a quadriceps muscle pain rating of 3 out of 10. Nine of the men completed follow-up testing which was identical except that the exercise consisted of 10min of cycling eliciting a pain rating of 5 out of 10. Caffeine had no effect compared to placebo on PPT (p≥0.15) or TPT (p≥0.41) 60min following ingestion and following exercise. PPT increased from 599±176kPa to 648±202kPa (p=0.009) and from 578±217kPa to 666±278kPa (p=0.01) following 15 and 10min of cycling, respectively. TPT increased from 46.2±2.9°C to 46.8±2.6°C (p=0.008) following the 15min exercise bout, but did not change (46.4±3.6°C vs. 46.8±3.3°C; p=0.24) following the shorter, higher intensity exercise bout. The results from this study indicate cycling exercise reduces pain sensitivity, especially to pressure stimuli. Caffeine ingestion did not alter the EIH response-suggesting adenosine may not play a prominent role in the EIH response in humans.


[Rapid and complete hematopoietic reconstitution following peripheral blood stem cell autotransplantation in a child with T cell acute lymphoblastic leukemia].
Rapid and complete hematopoietic reconstitution was achieved in a child with T cell acute lymphoblastic leukemia who was autografted with peripheral blood stem cells (PBSC). A large number of PBSC was collected by two courses of 3-4 hour-lasting lymphopheresis during early remission induced by the second-line chemotherapy and then cryopreserved in liquid nitrogen. A myeloid progenitor cell dose of 203 X 10(4) CFU-GM/kg body weight was reinfused to the patient following marrow-ablative chemotherapy (MCNU 600 mg/m2, cytosine arabinoside 6 g/m2, etoposide 300 mg/m2, cyclophosphamide 160 mg/kg). Neutrophil count reached 0.5 X 10(9)/l by day + 7 and platelet count reached 20 X 10(9)/l by day + 9. Thereafter, white blood cell count continued to increase and reached a maximum of 38 X 10(9)/l on day + 14. Thus, the rapid recovery of hematopoiesis minimised marrow aplasia-related risks. This approach of stem cell rescue operation can be applied to the treatment of children with cancer, who otherwise have no hope to be cured, as an alternative to bone marrow transplantation.


Interpersonal characteristics of male criminal offenders: personality, psychopathological, and behavioral correlates.
Interest in conceptualizing the interpersonal style of individuals who engage in serious antisocial behavior has increased in recent years. This study examines the personality, psychopathological, and behavioral correlates of interpersonal dominance and warmth, as operationalized via scales of the Personality Assessment Inventory (L. Morey, 2007), across several samples of male prison inmates (combined N = 1,062). Consistent with theory, multivariate analyses indicated that low warmth and, to a lesser extent, high dominance were associated with antisocial and paranoid traits, specifically, and externalizing-spectrum psychopathology more generally, whereas borderline traits and internalizing-spectrum psychopathology were uniquely associated with low interpersonal warmth. Among smaller subsamples of inmates followed prospectively, high dominance and, to a lesser extent, low warmth predicted general and aggressive institutional misconduct, whereas dominance uniquely predicted staff ratings of treatment noncompliance/failure. (PsycINFO Database Record (c) 2009 APA, all rights reserved).


Evaluation of the AMPLATZER vascular plug for embolization of peripheral vascular malformations associated with congenital heart disease.
The purpose of this study is to evaluate the recently FDA-approved AMPLATZER Vascular Plug in the embolization of vascular lesions associated with congenital heart disease (CHD). Fistulas and arteriovenous malformations have been occluded using various devices. The AMPLATZER Vascular Plug is a self-expandable, cylindrical device, used for embolization in the peripheral vasculature. A total of 84 vessels in 52 patients with CHD from 11 centers were occluded with 89 AMPLATZER Vascular Plugs, delivered through a coronary guide catheter in various vascular sites, including collaterals, pulmonary arterio-venous and coronary artery fistulas, transhepatic tracts, central shunts, patent ductus arteriosus (PDA), and excluded hepatic vein. Complete vessel occlusion was demonstrated within 10 min in 94% of patients. There was no device embolization, vascular disruption, or procedure-related complication. One vascular plug implanted in a large type C PDA required surgical removal followed by PDA ligation, after 5 weeks from successful implant because of significant residual flow through the device. The AMPLATZER Vascular Plug is an effective transcatheter occlusion device in the embolization of a wide variety of vascular lesions associated with CHD. Based on our early experience, caution should be used when considering the Vascular Plug as a closure device for large PDA.


Chronic kidney disease awareness, screening and prevention: rationale for the design of a public education program.
Design principles for a chronic kidney disease (CKD) screening program start with the general population at increased risk of CKD. Simple risk factor analysis demonstrates diabetes, hypertension, cardiovascular disease and older age as significant associated conditions. More comprehensive risk factor analysis shows only diabetes and hypertension as risk factors in people aged less than 50-60 years, and that anyone aged older than 50-60 years is at risk. Assessment of the relationship between CKD stage and cardiovascular risk factors shows early stage CKD to be associated with poor blood pressure control, which should be addressed. Other risk factors should be more completely assessed to determine if participants and their physicians are adequately addressing factors amenable to treatment to reduce high adverse event rates, premature death and progression to end-stage renal disease (ESRD). Such assessment is needed to reduce the high burden of ESRD on national health-care systems, which can only be addressed by early screening and active treatment.


Development of neurotransmitter metabolism in embryos of the leech Haementeria ghilianii.
We have investigated the development of neurotransmitter metabolism in embryos of the glossiphoniid leech Haementeria ghilianii. The neurotransmitter content of dissected embryonic tissues was measured by means of radioenzymatic assays, while the presence of neurotransmitters in individual identified neurons was detected by means of immunocytochemical and monoamine histofluorescence techniques. The capacity for synthesis of neurotransmitters was measured by incubating dissected embryonic tissues in radiolabeled neurotransmitter precursors. A specific neurotransmitter uptake system present in some neurons was detected by means of an autoradiographic technique. At an early stage of development of the nervous system, when most neurons are just beginning process outgrowth, the nerve cord acquires the capacity to synthesize ACh, 5-HT, and GABA from their immediate precursors, and contains ACh. Moreover, 5-HT-immunoreactive neurons and neurons that are capable of GABA uptake can be identified. Dopamine-containing neurons are first detected by their histofluorescence at a slightly later stage, after process outgrowth is under way. As development continues, the content of and capacity for synthesis of these neurotransmitters increase, as does the number of neurons capable of GABA uptake. During the earlier stages of development, ACh content exceeds 5-HT content, which in turn exceeds dopamine content. By the end of embryogenesis, however, 5-HT and dopamine contents have greatly increased relative to ACh content, with 5-HT content exceeding ACh content by a factor of 2. Of the neurotransmitters thus far studied, 5-HT is present in the highest amount in the juvenile and adult nerve cord. Our results indicate that in the development of the leech nervous system neurotransmitter metabolism is one of the first neuronal characters to differentiate and that the subsequent levels of the different neurotransmitters are differentially regulated.


Birefringent Properties of the Peripapillary Retinal Nerve Fiber Layer in Healthy and Glaucoma Subjects Analyzed by Polarization-Sensitive OCT.
To study the circumpapillary retinal nerve fiber layer (RNFL) birefringence (BIR) of early glaucoma and age-matched healthy eyes using polarization-sensitive optical coherence tomography (PS-OCT). In this prospective cross-sectional study, we compared virtual circular PS-OCT B-scans with a diameter of 3.5 mm centered on the optic disc (OD) acquired with a PS-OCT prototype (860 nm center wavelength). Early glaucoma was defined by the glaucomatous appearance of the OD and a pathologic visual field test with a mean deviation (MD) better than -6 dB. The main outcome parameters were BIR, RNFL-thickness (RNFL-T), and phase retardation (RET). The BIR value at each virtual A-scan position was the quotient of the RET measured at the inner segment/outer segment junction divided by the RNFL-T. The dataset comprised 49 early glaucoma patients (mean ± standard deviation [SD]: 64 ± 10 years) and 49 healthy control subjects (61 ± 9 years). Glaucomatous eyes showed a statistically significant lower BIR globally (mean ± SD: 0.108 ± 0.008°/µm vs. 0.112 ± 0.009°/µm, P = 0.033), superiorly (0.116 ± 0.017°/µm vs. 0.126 ± 0.013°/µm, P = 0.0001), and inferiorly (0.112 ± 0.011°/µm vs. 0.121 ± 0.011°/µm, P < 0.0001), and increased BIR in the temporal quadrant (0.088 ± 0.015°/µm vs. 0.078 ± 0.014°/µm, P = 0.0001) compared to healthy eyes. We report a reduced BIR of the RNFL in early perimetric glaucoma, which can be interpreted as a sign of loss or change of intracellular microtubules and may contribute to a better understanding of early disease development. Prospective longitudinal studies are needed to determine whether BIR is altered in pre-perimetric human glaucoma before RNFL-T decline.


Walnuts improve semen quality in men consuming a Western-style diet: randomized control dietary intervention trial.
We tested the hypothesis that 75 g of whole-shelled walnuts/day added to the Western-style diet of healthy young men would beneficially affect semen quality. A randomized, parallel two-group dietary intervention trial with single-blind masking of outcome assessors was conducted with 117 healthy men, age 21-35 yr old, who routinely consumed a Western-style diet. The primary outcome was improvement in conventional semen parameters and sperm aneuploidy from baseline to 12 wk. Secondary endpoints included blood serum and sperm fatty acid (FA) profiles, sex hormones, and serum folate. The group consuming walnuts (n = 59) experienced improvement in sperm vitality, motility, and morphology, but no change was seen in the group continuing their usual diet but avoiding tree nuts (n = 58). Comparing differences between the groups from baseline, significance was found for vitality (P = 0.003), motility (P = 0.009), and morphology (normal forms; P = 0.04). Serum FA profiles improved in the walnut group with increases in omega-6 (P = 0.0004) and omega-3 (P = 0.0007) but not in the control group. The plant source of omega-3, alpha-linolenic acid (ALA) increased (P = 0.0001). Sperm aneuploidy was inversely correlated with sperm ALA, particularly sex chromosome nullisomy (Spearman correlation, -0.41, P = 0.002). Findings demonstrated that walnuts added to a Western-style diet improved sperm vitality, motility, and morphology.


[Circulatory mechanical support. State of the art].
Despite advances in medical treatment, the prognosis of advanced heart failure remains poor. The number of hospitalizations for heart failure exacerbations continues to increase and most patients will ultimately die of complications related to heart failure. Implantable left ventricular assist devices (LVAD) are currently in use throughout the world with increasing frequency. This paper is a comprehensive review about mechanical support, focusing on a general description of the different LVAD, complications and mortality.


Serotonin 5-HT2 receptor interactions with dopamine function: implications for therapeutics in cocaine use disorder.
Cocaine exhibits prominent abuse liability, and chronic abuse can result in cocaine use disorder with significant morbidity. Major advances have been made in delineating neurobiological mechanisms of cocaine abuse; however, effective medications to treat cocaine use disorder remain to be discovered. The present review will focus on the role of serotonin (5-HT; 5-hydroxytryptamine) neurotransmission in the neuropharmacology of cocaine and related abused stimulants. Extensive research suggests that the primary contribution of 5-HT to cocaine addiction is a consequence of interactions with dopamine (DA) neurotransmission. The literature on the neurobiological and behavioral effects of cocaine is well developed, so the focus of the review will be on cocaine with inferences made about other monoamine uptake inhibitors and releasers based on mechanistic considerations. 5-HT receptors are widely expressed throughout the brain, and several different 5-HT receptor subtypes have been implicated in mediating the effects of endogenous 5-HT on DA. However, the 5-HT2A and 5-HT2C receptors in particular have been implicated as likely candidates for mediating the influence of 5-HT in cocaine abuse as well as to traits (e.g., impulsivity) that contribute to the development of cocaine use disorder and relapse in humans. Lastly, new approaches are proposed to guide targeted development of serotonergic ligands for the treatment of cocaine use disorder.


Immunoblot analysis of Toxoplasma gondii antigens by human immunoglobulins G, M, and A antibodies at different stages of infection.
The Toxoplasma gondii antigenic components eliciting the immunoglobulin G (IgG), IgM, and IgA antibody responses were studied by using follow-up sera from a laboratory worker who developed an acute glandular toxoplasmosis after an accidental infection with the protozoa. IgG toxoplasma antibodies reacted with multiple components over a wide molecular weight range from 6,000 to 150,000. In contrast, IgM toxoplasma antibodies reacted predominantly with polypeptides of 6, 25, and 35 kilodaltons, which might be useful in new diagnostic procedures. The general pattern of antigenic components in the IgA toxoplasma antibody response closely resembled that in the IgM response, even though some characteristic features were constantly observed. The possibility that the restricted IgM and IgA antibody responses relate to the pathogenetic events in human toxoplasmosis is considered.


Recognition of T cell epitopes and lymphokine secretion by rye grass allergen Lolium perenne I-specific human T cell clones.
T cell lines (TCL) and CD4+ T cell clones (TCC) with specificity for the rye grass allergen Lolium perenne (Lol p) I were isolated from the blood of nine donors, six having active atopic disease, two being in remission, and one having IgE anti-Lol pI Abs but not atopic disease. The T cell epitopes of Lol pI were determined by TCLs and TCCs reactivity with 23 overlapping, 20 amino acid-long peptides spanning the entire length of the 230 amino acid-long allergen. In addition, the Th subsets (Th1, Th2, Th0, Thp) were determined by measuring IL-2, IFN-gamma, and IL-4 in the supernatants of TCC activated with Lol pI and irradiated APC. TCC from individuals from which a large panel of clones were obtained from 10(5) PBMC initial cultures recognized multiple peptides (5-9) and 23 overlapping peptides a total of 16 were recognized by at least one TCC from one of the patients. These 16 peptides were derived from all areas of the Lol pI molecule, indicating the ability of human Th cells to recognize many peptide epitopes on Lol pI. Although no clear cut immunodominant peptides were detected, T cell clones of 50% of the patients reacted with peptide 191-210. There was no correlation between peptide epitope reactivity and lymphokine secretion pattern of the TCC. Of 12 TCC obtained from six patients with active atopic disease, four (33%) were of Th1, five (42%) of Th2, one (8%) of Thp, and two (17%) of Th0 type. Of 14 TCCs isolated from three atopic donors in remission, five (36%) were of Th1, three (21%) of Th2, four (29%) of Thp, and two (14%) of Th0 type. The data demonstrate that T cells from rye grass pollen allergic patients can recognize multiple peptide epitopes on Lol pI scattered over the entire molecule. No correlation existed between epitope reactivity and lymphokine secretion pattern of the TCC.


Trade-Offs between Error, Speed, Noise, and Energy Dissipation in Biological Processes with Proofreading.
High accuracy of major biological processes relies on the ability of the participating enzymatic molecules to preferentially select the correct substrate from a pool of chemically similar substrates by activating the so-called proofreading mechanisms. While the importance of such mechanisms is widely accepted, it is still unclear how evolution has optimized the biological systems with respect to certain characteristic properties. Here, using a discrete-state stochastic framework with a first-passage analysis, we theoretically investigate trade-offs between four characteristic properties of enzymatic systems, namely, error, speed, noise, and energy dissipation. Specifically, two fundamental biological processes are examined, i.e., DNA replication in the T7 bacteriophage and tRNA selection during protein translation in Escherichia coli. Notably, all of the characteristic properties cannot be completely optimized at the same time due to trade-offs between them. To understand the relative importance of the computed quantities to the enzymatic functionality, we introduce a new quantitative metric to rank the properties. The results demonstrate that the reaction speed is the principal characteristic property that evolution optimizes in both enzymatic systems and that the energy dissipation comes in second. In addition, the error and the noise are always ranked third and fourth, respectively, regardless of the system considered. Physicochemical arguments to explain these observations are presented.


Diethyl 1,8-bis-(4-methyl-phen-yl)-11-oxatricyclo-[18.104.22.168(2,7)]undeca-2,4,6-triene-9,10-dicarboxyl-ate.
The title compound, C30H30O5, is the Diels-Alder adduct from 1,3-diphenyl-benzo[c]furan and diethyl maleate. The mol-ecule comprises a fused tricyclic system containing two five-membered rings, which are in envelope conformations with the O atom at the flap, and a six-membered ring adopting a boat conformation. The dihedral angle between the 4-methyl-phenyl substituents in the 1- and 8-positions is 62.1 (1)°. The ethyl group of one ester group and the eth-oxy group of the other ester group are disordered over two sets of sites, with occupancy ratios of 0.43 (2):0.57 (2) and 0.804 (7):0.196 (7), respectively. In the crystal, inversion dimers are formed through pairs of C-H⋯O inter-actions.


Weighting functions and data truncation in the fitting of multi-exponential functions.
Artificial data were simulated by using two-exponential functions and normally distributed pseudo-random numbers. The variation corresponded to either constant or relative variance for the data error. These data were used to test (i) three different weighting functions and (ii) the effect of data truncation on the precision of estimating the parameters of two-exponential functions.


Identification of mosquito larvicidal bacterial strains isolated from north Sinai in Egypt.
In the present study, two of the most toxic bacterial strains of Bacillus sphaericus against mosquito were identified with the most recent genetic techniques. The PCR product profiles indicated the presence of genes encoding Bin A, Bin B and Mtx1 in all analyzed strains; they are consistent with protein profiles. The preliminary bioinformatics analysis of the binary toxin genes sequence revealed that the open reading frames had high similarities when matched with nucleotides sequence in the database of other B. sphaericus strains. The biological activity of B. sphaericus strains varied according to growing medium, and cultivation time. The highest yield of viable counts, spores and larvicidal protein were attained after 5 days. Poly (P) medium achieved the highest yield of growth, sporulation, protein and larvicidal activity for all tested strains compared to the other tested media. The larvicidal protein produced by local strains (B. sphaericus EMCC 1931 and EMCC 1932) in P medium was more lethal against the 3rd instar larvae of Culex pipiens than that of reference strains (B. sphaericus 1593 and B. sphaericus 2297). The obtained results revealed that P medium was the most effective medium and will be used in future work in order to optimize large scale production of biocide by the locally isolated Bacillus sphaericus strains.


Serotonergic Psychedelics in Neural Plasticity.
Psychedelics, compounds that can induce dramatic changes in conscious experience, have been used by humans for centuries. Recent studies have shown that certain psychedelics can induce neural plasticity by promoting neurite growth and synapse formation. In this review, we focus on the role of classical serotonergic psychedelics in neural plasticity and discuss its implication for their therapeutic potentials.


Cognitive-behavioral therapy for chronic insomnia.
Psychological and behavioral therapies should be considered the first line treatment for chronic insomnia. Although cognitive behavioral therapy for insomnia (CBT-I) is considered the standard of care [1], several monotherapies, including sleep restriction therapy, stimulus control therapy, and relaxation training are also recommended in the treatment of chronic insomnia [2]. CBT-I is a multimodal intervention comprised of a combination of behavioral (eg, sleep restriction, stimulus control) and cognitive therapy strategies, and psychoeducation delivered in 4 to 10 weekly or biweekly sessions [3]. Given that insomnia is thought to be maintained by an interaction between unhelpful sleep-related beliefs and behaviors, the goal of CBT-I is to modify the maladaptive cognitions (eg, worry about the consequences of poor sleep), behaviors (eg, extended time in bed), and arousal (ie, physiological and mental hyperarousal) perpetuating the insomnia. CBT-I is efficacious when implemented alone or in combination with a pharmacologic agent. However, because of the potential for relapse upon discontinuation, CBT-I should be extended throughout drug tapering [4]. Although the treatment options should be guided by the available evidence supporting both psychological therapies and short-term hypnotic treatment, as well as treatment feasibility and availability, treatment selection should ultimately be guided by patient preference [5]. Despite its widespread use among treatment providers [6], the use of sleep hygiene education as a primary intervention for insomnia should be avoided. Sleep hygiene may be a necessary, but insufficient condition for promoting good sleep and should be considered an adjunct to another empirically supported treatment.


Angiotensin II receptors in rabbit renal preglomerular vessels.
Controversy exists regarding the specific sites within the renal microcirculation affected by angiotensin II (ANG II). Under some conditions, ANG II can elicit direct vasoconstrictor responses in the preglomerular vessels and efferent arterioles. These experiments were designed to evaluate the binding of 125I-ANG II in preglomerular vessels. Arcuate and interlobular arteries, with attached proximal segments of afferent arterioles, were microdissected from rabbit renal cortexes. A membrane preparation was obtained from the pooled freshly dissected vessels and utilized in an ANG II radioreceptor assay on the same day. Binding site concentrations [N] and dissociation constants [KD] were obtained by Scatchard analyses of binding inhibition data. Specific binding was saturable and reversible. The dissociation of bound ANG II was enhanced in the presence of a nonhydrolyzable analogue of GTP. Linear Scatchard plots were obtained, indicating the presence of a single class of high-affinity binding sites. The KD and N are similar to those for ANG II receptors in extrarenal vascular tissue. The order of binding inhibition potencies of ANG analogues was [Sar1,Ile8]-ANG II much greater than [Sar1,Ala8]ANG II = ANG II = ANG III much greater than ANG I, which is consistent with in vivo observations of the effects of these analogues on renal blood flow. The binding inhibition potencies of ANG III and [Sar1,Ile8]ANG II were greater in renal compared with reported values for extrarenal vasculature and rabbit glomeruli. Furthermore, there were no differences in ANG II receptor parameters in preglomerular vessels obtained from pregnant and nonpregnant rabbits.(ABSTRACT TRUNCATED AT 250 WORDS)


Structure and function in galactosyltransferase. Sequence locations of alpha-lactalbumin binding site, thiol groups, and disulfide bond.
The region(s) of bovine galactosyltransferase that interacts with the lactose synthase regulatory protein alpha-lactalbumin was investigated using trace 3H acetylation to probe the effects of alpha-lactalbumin on the reactivities of the individual amino groups of galactosyltransferase. In the presence of Mn2+, alpha-lactalbumin was found to reduce the reactivities of lysines 93 and 181 and to increase the reactivities of one or more of lysines 230, 237, and 241. The addition of N-acetylglucosamine (20 mM), which enhances complex formation between the two proteins, did not significantly alter the pattern of perturbation. These results indicate that the NH2-terminal region of the catalytic domain of galactosyltransferase, and possibly part of the proline-rich "stem" region, is affected by the association with alpha-lactalbumin and is therefore implicated in the binding of acceptor substrates. In a separate study only cysteines 176, 266, and 342 of galactosyltransferase were found to react with [3H]iodoacetic acid under denaturing conditions. From their lack of reactivity it is deduced that the remaining two cysteines, residues 134 and 247, are joined in a disulfide linkage. From these results and those of a previous study of UDP-galactose binding (Yadav, S., and Brew, K. (1990) J. Biol. Chem. 265, 14163-14169) it appears that the soluble form of galactosyltransferase is composed of two domains, the NH2-terminal 150 residues containing the Cys134-Cys247 disulfide bond, which functions in alpha-lactalbumin and acceptor binding, and the COOH-terminal region, which is involved in UDP-galactose binding.


Management of vertebral column fractures in dogs and cats: 211 cases (1977-1985).
Hospital records and radiographs of 211 dogs and cats with vertebral column fractures or luxations evaluated at the University of Tennessee Veterinary Teaching Hospital between April 1977 and September 1985 were reviewed. After neurologic examination, status of the animal was graded on a scale of 1 to 8. Decision to treat each animal either medically or surgically was based on the extent and type of injury, neurologic signs, veterinarian's experience, and owner's wishes. After treatment, neurologic status was evaluated on the aforementioned scale and differences in the outcome of treatment were determined between surgically and medically treated groups, relative to initial neurologic status and location of the fracture. Surgically treated animals had pretreatment mean (+/- SD) neurologic status (3.71 +/- 1.35) that was slightly worse (P = 0.0079) than that of medically managed cases (5.16 +/- 1.48). Animals of the surgically treated group improved significantly (P = 0.0122) more than did those of the medically treated group but after treatment, significant differences in neurologic status were not evident between surgically (6.67 +/- 1.49) and medically (7.07 +/- 1.24) treated animals. Medically treated animals required substantially longer to reach optimal neurologic status, but the average hospital stay was nearly twice as long for the surgically treated animals (13.5 days), compared with those treated medically (7.1 days). Animals with thoracic vertebral fractures had mean neurologic status that was worse than that in animals with cervical vertebral fractures (P = 0.0109). After either medical or surgical treatment, neurologic status did not differ among animals with cervical, thoracic, or lumbar vertebral fractures.(ABSTRACT TRUNCATED AT 250 WORDS)


Activation of NF-κB by the Kaposi's sarcoma-associated herpesvirus K15 protein involves recruitment of the NF-κB-inducing kinase, IκB kinases, and phosphorylation of p65.
Kaposi's sarcoma herpesvirus (KSHV) (or human herpesvirus 8) is the cause of Kaposi's sarcoma, primary effusion lymphoma (PEL), and the plasma cell variant of multicentric Castleman's disease (MCD). The transmembrane K15 protein, encoded by KSHV, has been shown to activate NF-κB and the mitogen-activated protein kinases (MAPKs) c-jun-N-terminal kinase (JNK) and extracellular signal-regulated kinase (Erk) as well as phospholipase C gamma (PLCγ) and to contribute to KSHV-induced angiogenesis. Here we investigate how the K15 protein activates the NF-κB pathway. We show that activation of NF-κB involves the recruitment of NF-κB-inducing kinase (NIK) and IKK α/β to result in the phosphorylation of p65/RelA on Ser536. A K15 mutant devoid in NIK/IKK recruitment fails to activate NF-κB but remains proficient in the stimulation of both NFAT- and AP1-dependent promoters, showing that the structural integrity of the mutant K15 protein has not been altered dramatically. Direct recruitment of NIK represents a novel way for a viral protein to activate and manipulate the NF-κB pathway. KSHV K15 is a viral protein involved in the activation of proinflammatory and angiogenic pathways. Previous studies reported that K15 can activate the NF-κB pathway. Here we show the molecular mechanism underlying the activation of this signaling pathway by K15, which involves direct recruitment of the NF-κB-inducing kinase NIK to K15 as well as NIK-mediated NF-κB p65 phosphorylation on Ser536. K15 is the first viral protein shown to activate NF-κB through direct recruitment of NIK. These results indicate a new mechanism whereby a viral protein can manipulate the NF-κB pathway.


Brief Review of Chloroquine and Hydroxychloroquine Toxicity and Management.
As of April 21, 2020, more than 2.5 million cases of coronavirus disease 2019 (COVID-19), caused by the SARS-CoV-2 virus, have been reported in 210 countries and territories, with the death toll at 171,810. Both chloroquine and hydroxychloroquine have gained considerable media attention as possible therapies, resulting in a significant surge in demand. In overdose, both medications can cause severe, potentially life-threatening effects. Here, we present a brief overview of the pharmacology of chloroquine and hydroxychloroquine, manifestations of toxicity, and treatment considerations.


Methods for perfusing the male reproductive tract: models for studying drug and hormone metabolism.
Methods for perfusing rat testis and accessory sex organs in situ by recirculating artificial medium to the hemicorpus preparation are described. The advantages and limitations of this system for studying the male reproductive tract were examined. The preparation was then used to study the uptake of androgens into the nuclei of caput epididymis, ventral prostate, seminal vesicle and testis. The accumulation of dihydrotestosterone and accessory sex organ nuclei was saturable and inhibited by perfusion of excess testosterone or cyproterone acetate. By contrast, testosterone was the major nuclear androgen in the testis of mature hypophysectomized preparations perfused with testosterone. In all parts of the reproductive tract, (3)H-nuclear androgens were associated with 3S, salt-extractable macromolecules within the properties of androgen receptors.The hemicorpus preparation was extensively compared with two techniques (selective and isolated) for perfusing the testis directly. Of these two procedures, the isolated method was superior when only the testes were studied. However, the hemicorpus preparation offers the advantage of studying testes along with the remainder of the male reproductive tract. A variety of observations suggest that these perfusion procedures will be useful for the study of drug as well as hormone metabolism and mechanism of action.


[A study of maxillary and mandibular vasculature in relation to orthognathic surgery].
Angiography and Chinese ink perfusion were performed in 14 cadavers. It was found that the origins of arterial blood supply to maxilla and mandible in human beings are multiple. Normally, the maxilla and mandible receive not only the centrifugal blood from the alveolar arteries, but also the centripetal blood from the surrounding mucoperiosteum. This feature of jaw bone's blood supply is the important anatomic basis for the establishment of blood compensation to the moved bone segments which are nourished only by attached mucoperiostea after osteotomies have cut off the blood supply of alveolar arteries in orthognathic surgery.


Biochemical characterization of Sfh-I, a subclass B2 metallo-beta-lactamase from Serratia fonticola UTAD54.
The subclass B2 metallo-β-lactamase (MBL) Sfh-I from Serratia fonticola UTAD54 was cloned and overexpressed in Escherichia coli. The recombinant protein binds one equivalent of zinc, as shown by mass spectrometry, and preferentially hydrolyzes carbapenem substrates. However, compared to other B2 MBLs, Sfh-I also shows limited hydrolytic activity against some additional substrates and is not inhibited by a second equivalent of zinc. These data confirm Sfh-I to be a subclass B2 metallo-β-lactamase with some distinctive properties.


The Impact of Somatostatin Receptor-Directed PET/CT on the Management of Patients with Neuroendocrine Tumor: A Systematic Review and Meta-Analysis.
Somatostatin receptor (SSTR) imaging is widely used for guiding the management of neuroendocrine tumor (NET) patients. 68Ga-DOTATATE approval by the U.S. Food and Drug Administration has triggered widespread clinical interest in SSTR PET/CT throughout the United States. Here, we performed a systematic review and meta-analysis to evaluate the impact of SSTR PET/CT on the management of patients with NETs. Methods: A comprehensive literature search was performed using The National Center for Biotechnology Information PubMed online database, applying the following key words: "management" AND "PET" AND "neuroendocrine". Fourteen of 190 studies were deemed suitable based on the following inclusion criteria: original research, cohort study, number of patients 10 or more, and reported change in management after SSTR PET/CT. Change in management across studies was determined by a random-effects model. Results: A total of 1,561 patients were included. Overall, change in management occurred in 44% (range, 16%-71%) of NET patients after SSTR PET/CT. In 4 of 14 studies, SSTR PET/CT was performed after an 111In-Octreotide scan. In this subgroup, additional information by SSTR PET/CT led to a change in management in 39% (range, 16%-71%) of patients. Seven of 14 studies differentiated between inter- and intramodality changes, with most changes being intermodality (77%; intramodality, 23%). Conclusion: The management was changed in more than one third of patients undergoing SSTR PET/CT even when performed after an 111In-Octreotide scan. Intermodality changes were 3 times more likely than intramodality changes, underlining the clinical impact of SSTR PET/CT.


PSD-95 is essential for hallucinogen and atypical antipsychotic drug actions at serotonin receptors.
Here, we report that postsynaptic density protein of 95 kDa (PSD-95), a postsynaptic density scaffolding protein, classically conceptualized as being essential for the regulation of ionotropic glutamatergic signaling at the postsynaptic membrane, plays an unanticipated and essential role in mediating the actions of hallucinogens and atypical antipsychotic drugs at 5-HT(2A) and 5-HT(2C) serotonergic G-protein-coupled receptors. We show that PSD-95 is crucial for normal 5-HT(2A) and 5-HT(2C) expression in vivo and that PSD-95 maintains normal receptor expression by promoting apical dendritic targeting and stabilizing receptor turnover in vivo. Significantly, 5-HT(2A)- and 5-HT(2C)-mediated downstream signaling is impaired in PSD-95(null) mice, and the 5-HT(2A)-mediated head-twitch response is abnormal. Furthermore, the ability of 5-HT(2A) inverse agonists to normalize behavioral changes induced by glutamate receptor antagonists is abolished in the absence of PSD-95 in vivo. These results demonstrate that PSD-95, in addition to the well known role it plays in scaffolding macromolecular glutamatergic signaling complexes, profoundly modulates metabotropic 5-HT(2A) and 5-HT(2C) receptor function.


Ligamentoplasty using the ulnotriquetral ligament with wafer procedure for chronic triangular fibrocartilage complex foveal tear: Short-tem outcome.
We developed a new ligamentoplasty technique using the ulnotriquetral ligament (UTL) via a palmar approach for cases with dorsal instability of the ulna due to a foveal tear of the triangular fibrocartilage complex (TFCC). The study aimed to evaluate the clinical outcomes of this procedure. We retrospectively analyzed 19 cases with foveal tears of TFCC who underwent the ligamentoplasty using UTL. We harvested the UTL from the triquetrum as a pedicle graft, avoiding damage of the connection between the base of the UTL and palmar radioulnar ligament (PRUL) as possible. The graft was flipped proximally and pulled out through the bone tunnel from the ulnar fovea to the ulnar neck, then fixed to the fovea using a tenodesis screw. The function of the TFCC can be reproduced by pulling out the UTL along with the PRUL through the ulnar fovea and re-establishing the PRUL tension. Clinical results were evaluated at least 12 months following surgery. The preoperative and postoperative wrist pain level on a neumerical rating scale (NRS), the radioulnar joint (DRUJ) ballottement test, the range of motion, the grip strength, and the Patient-rated wrist evaluation (PRWE) scores were evaluated. In all cases, the DRUJ ballottement test became negative at the final follow up compared with the normal side on manual examination. The pain levels significantly decreased as reflected by a decrease in NRS from 6.8 to 2.5 at final follow up (p < 0.01). The range of pronation/supination motion was improved from 149° to 157°. The mean PRWE score significantly decreased from 52.1 to 22.8 (p < 0.01). The postoperative pain level on the NRS scale of the three patients poorly improved. One case of the three with the 3 mm positive ulnar variance needed additional ulnar shaft shortening using a plate seven months after the primary procedure. The ligamentoplasty using UTL efficiently restored the ulnar palmar stability in all 19 cases and significantly decreased the wrist pain and the PRWE scores. Clinical, retrospective study.


Molecular mechanisms of acid-base sensing by the kidney.
A major function of the kidney is to collaborate with the respiratory system to maintain systemic acid-base status within limits compatible with normal cell and organ function. It achieves this by regulating the excretion and recovery of bicarbonate (mainly in the proximal tubule) and the secretion of buffered protons (mainly in the distal tubule and collecting duct). How proximal tubular cells and distal professional proton transporting (intercalated) cells sense and respond to changes in pH, bicarbonate, and CO(2) status is a question that has intrigued many generations of renal physiologists. Over the past few years, however, some candidate molecular pH sensors have been identified, including acid/alkali-sensing receptors (GPR4, InsR-RR), kinases (Pyk2, ErbB1/2), pH-sensitive ion channels (ASICs, TASK, ROMK), and the bicarbonate-stimulated adenylyl cyclase (sAC). Some acid-sensing mechanisms in other tissues, such as CAII-PDK2L1 in taste buds, might also have similar roles to play in the kidney. Finally, the function of a variety of additional membrane channels and transporters is altered by pH variations both within and outside the cell, and the expression of several metabolic enzymes are altered by acid-base status in parts of the nephron. Thus, it is possible that a master pH sensor will never be identified. Rather, the kidney seems equipped with a battery of molecules that scan the epithelial cell environment to mount a coordinated physiologic response that maintains acid-base homeostasis. This review collates current knowledge on renal acid-base sensing in the context of a whole organ sensing and response process.


Ultrasound analysis of tongue contour for the sound [j] in adults and children.
Analyze and compare the mean tongue contours and articulatory gestures in the production of the sound [j] in adults and children with typical and atypical speech development. The children with atypical development presented speech sound disorders. The diagnosis was determined by speech assessments. The study sample was composed of 90 individuals divided into three groups: 30 adults with typical speech development aged 19-44 years (AT), 30 children with typical speech development (CT), and 30 children with speech sound disorders, named as atypical in this study, aged four years to eight years and eleven months (CA). Ultrasonography assessment of tongue movements was performed for all groups. Mean tongue contours were compared between three groups in different vocalic contexts following the sound [j]. The maximum elevation of the tongue tip was considered for delimitation of gestures using the Articulate Assistant Advanced (AAA) software and images in sagittal plane/Mode B. The points that intercepted the language curves were analyzed by the statistical tool R. The graphs of tongue contours were obtained adopting a 95% confidence interval. After that, the regions with significant statistical differences (p<0.05) between the CT and CA groups were obtained. The mean tongue contours demonstrated the gesture for the sound [j] in the comparison between typical and atypical children. For the semivowel [j], there is an articulatory gesture of tongue and dorsum towards the center of the hard palate, with significant differences observed between the children. The results showed differences between the groups of children regarding the ability to refine articulatory gestures.


Using Spatial Analysis to Inform Community Immunization Strategies.
Recent pertussis outbreaks in the United States suggest our response to local disease outbreaks (eg, vaccine-preventable Bordetella pertussis) may benefit from understanding and applying spatial analytical methods that use data from immunization information systems at a subcounty level. A 2012 study on Denver, CO, residents less than 19 years of age confirmed pertussis cases and immunization information system records were geocoded and aggregated to the census tract (CT) level. An algorithm assessed whether individuals were up-to-date (UTD) for pertussis vaccines. Pearson, Spearman, and Kendall correlations assessed relations between disease incidence and pertussis vaccine coverage. Using spatial analysis software, disease incidence and UTD rates were spatially weighted, and smoothed. Global and local autocorrelations based on univariate Moran's I spatial autocorrelation statistics evaluated whether a CT's rate belong to a cluster based on incidence or UTD measures. Overall disease incidence rate was 116.8/100 000. Assessment of pertussis vaccination coverage was available for 90% of the population. Among 134 672 Denver residents less than 19 years old, 103 496 (77%) were UTD for pertussis vaccines. Raw correlation coefficients showed weak relationships between incidence and immunization rates due to the presence of outliers. With geospatial and clustering analysis, estimates and correlation coefficients were improved with statistically significant Moran's I values for global and local autocorrelations rejecting the null hypothesis that incidence or UTD rates were randomly distributed. With evidence indicating the presence of clusters, smoothed and weighted disease incidence and UTD rates in 144 CTs identified 21 CTs (15%) for potential public health intervention. Correlation of raw disease incidence and vaccine UTD rates in subcounty regions showed limited association, providing limited information for decision making. By assessing for clusters using spatial analysis methods, we identified CTs with higher incidence and lower immunization coverage for targeted public health interventions.


[Microbiological procedures for diagnosing mycoses and for antifungal susceptibility testing].
Fungal infections are a diagnostic and therapeutic problem of increasing concern due to the frequency and severity of disseminated infection in immunocompromised patients. Culture-based methods are characteristically slow and have poor sensitivity; hence, other methods, based on the detection of fungus-specific genetic, antigenic and metabolic components are being developed to enable early diagnosis and specific treatment. Moreover, reproducible antifungal susceptibility methods that can be adapted for use in clinical laboratories have been standardized to allow in vitro detection of resistance, which correlates with a less favorable clinical outcome. In this paper we review the main microbiological procedures available for the diagnosis of fungal infections and for antifungal susceptibility testing.


A Simple Method for in situ Quantification of Cells on Carriers.
The technology of cell carriers was developed as a response to the need for high cell density to enable higher production levels in cell-based production processes. To follow the production process, quantifying the number of cells on these carriers is required, as well as tracking their viability and proliferation. However, owing to various carriers' unique structures, tracking the cells is challenging using current traditional assays that were originally developed for monolayers of adherent cells. The current "gold standard" method is counting cell nuclei, which is tedious and counts both live and dead cells. A few other techniques have been developed, but they are all specific to a carrier type and involve specialized equipment. Here, we describe a broad ranging method for counting cells on carriers. The method is based on the Alamar blue dye, a well-known, common marker for cell activity. No separation of the cells from the carriers is needed, nor is any specialized equipment. The method is simple and rapid, and provides comprehensive details necessary for control of production processes in cells. This method can be easily implemented in any cell-based process and other unique platforms for measuring growth of cells. Graphic abstract: Schematic of the in situ quantification method.


Memory: Looking back and looking forward.
This review brings together past and present achievements in memory research, ranging from molecular to psychological discoveries. Despite some false starts, major advances include our growing understanding of learning-related neural plasticity and the characterisation of different classes of memory. One striking example is the ability to reactivate targeted neuronal ensembles so that an animal will seemingly re-experience a particular memory, with the further potential to modify such memories. Meanwhile, human functional imaging studies can distinguish individual episodic memories based on voxel activation patterns. While the hippocampus continues to provide a rich source of information, future progress requires broadening our research to involve other sites. Related challenges include the need to understand better the role of glial-neuron interactions and to look beyond the synapse as the sole site of experience-dependent plasticity. Unmet goals include translating our neuroscientific knowledge in order to optimise learning and memory, especially among disadvantaged populations.


Delivery of femtolitre droplets using surface acoustic wave based atomisation for cryo-EM grid preparation.
Cryo-Electron Microscopy (cryo-EM) has become an invaluable tool for structural biology. Over the past decade, the advent of direct electron detectors and automated data acquisition has established cryo-EM as a central method in structural biology. However, challenges remain in the reliable and efficient preparation of samples in a manner which is compatible with high time resolution. The delivery of sample onto the grid is recognized as a critical step in the workflow as it is a source of variability and loss of material due to the blotting which is usually required. Here, we present a method for sample delivery and plunge freezing based on the use of Surface Acoustic Waves to deploy 6-8 µm droplets to the EM grid. This method minimises the sample dead volume and ensures vitrification within 52.6 ms from the moment the sample leaves the microfluidics chip. We demonstrate a working protocol to minimize the atomised volume and apply it to plunge freeze three different samples and provide proof that no damage occurs due to the interaction between the sample and the acoustic waves.


[Inhibition of bacterial agmatinase by substrate analogs].
Activity of agmatinase (EC 220.127.116.11) from Proteus vulgaris was studied in presence of guanidine derivatives of agmatine as substrates. The guanidine derivatives, containing carboxyl group, did not interact with the enzyme. An inhibitory effect developed if the carboxyl group was esterified. For exhibition of the effect the length of hydrocarbon radical in ligand and presence of hydrophobic groups were important. One of the most effective inhibitor was N-isoamylene agmatine (K1=0.001 M). Compounds, containing an amino- or guanidine group at the position opposite to the guanidine end and possessing the hydrocarbon chain not less that C4 were shown to be substrates of agmatinase.


Vaccines, reverse vaccinology, and bacterial pathogenesis.
Advances in genomics and innovative strategies such as reverse vaccinology have changed the concepts and approaches to vaccine candidate selection and design. Genome mining and blind selection of novel antigens provide a novel route to investigate the mechanisms that underpin pathogenesis. The resulting lists of novel candidates are revealing new aspects of pathogenesis of target organisms, which in turn drives the rational design of optimal vaccine antigens. Here we use the discovery, characterization, and exploitation of fHbp, a vaccine candidate and key virulence factor of meningococcus, as an illustrative case in point. Applying genomic approaches to study both the pathogen and host will ultimately increase our fundamental understanding of pathogen biology, mechanisms responsible for the development of protective immunity, and guide next-generation vaccine design.


[Control of the quality of therapeutic process in hospital departments].
It is recommended to use the following indices for assessing the quality of hospital treatment: average hospital stay, the percent of repeated admissions with one and the same disease, the percent of discrepancy between clinical and postmortem diagnosis with subsequent expert evaluation of each case of death. The departments of surgery also find it necessary to take into account the frequency of postoperative complications, postoperative lethality and average hospital stay of operated patients. The well-founded complaints per 100 hospitalized patients can also be considered as indices.


Transcriptomic changes associated with DKK4 overexpression in pancreatic cancer cells detected by RNA-Seq.
The promotion of tumor development by Dickkopf 4 (DKK4) is receiving increased attention. However, the association between DKK4 and pancreatic cancer remains unclear. DKK4 expression was measured in pancreatic ductal adenocarcinoma tissues using qRT-PCR and immunohistochemistry. A DKK4-overexpressing pancreatic cancer cell line was established, and the differentially expressed genes (DEGs) that were induced by DKK4 were identified using transcriptome sequencing. The association between the identified DEGs and pancreatic cancer was assessed using gene ontology (GO), pathway analysis, pathway interaction networks, differentially expressed gene interaction network analysis, and co-expression gene networks. Finally, the accuracy of the analyses was validated using serial paraffin and frozen sections of clinical samples. DKK4 is highly expressed in pancreatic cancer tissues. DEGs of overexpression DKK4 of PANC-1 are mostly upregulated. GO and pathway analysis showed that DKK4 are associated with tumor and organ development and immune inflammation. The mitogen-activated protein kinase (MAPK) signaling pathway was the main signal transduction pathway that showed significant enrichment in overexpression DKK4 of PANC-1. The results of GO, pathway analyses, and differentially expressed gene interaction network identified genes that are closely associated with tumor development, including MAPK3, PIK3R3, VAV3, JAG1, and Notch3. The immunohistochemistry and immunofluorescence results suggested that DKK4 is co-expressed with MAPK3 and VAV3 in pancreatic cancer tissues. The results presented here show for the first time that DKK4 is highly expressed in pancreatic cancer tissues. Bioinformatics analysis of a DKK4-overexpressing of PANC-1 identified several oncogenes that are closely associated with tumors, and the MAPK signaling pathway is the core signal transduction pathway. DKK4 can be co-expressed with MAPK3 and VAV3 in pancreatic ductal adenocarcinoma tissues. Thus, DKK4 may have function on the development and progression of pancreatic cancer.


A novel near-infrared fluorescent probe with an improved Stokes shift for specific detection of Hg2+ in mitochondria.
The mercury ion (Hg2+), one of the most notorious heavy metal ions, not only causes environmental pollution, but also endangers human health. There is evidence that Hg2+ tends to accumulate in the mitochondria and to induce apoptosis. However, mitochondria-targeted near-infrared (NIR) fluorescent probes with large Stokes shifts are still scarcely described for the specific detection of Hg2+. In this work, a novel near-infrared fluorescent probe JRQNS with a large Stokes shift (78 nm) was reported, and applied for sensitive and specific detection of Hg2+ in mitochondria by incorporating an additional amine group with fused rings to rhodamine dyes to enhance the electron donating ability of amine groups. As expected, the probe exhibited high selectivity and sensitivity to Hg2+ with a detection limit as low as 1.5 nM and fast response times (3 min), revealing that JRQNS could be used as a practical probe for quantitative detection of Hg2+ in real-time. Importantly, JRQNS can be used as an efficient organelle-targeting probe for imaging Hg2+ in the mitochondria of living cells, and thus detect Hg2+ in real-time there. The application of the probe for its selective localization in mitochondria along with the nanomolar level of limit of detection to Hg2+ ions provided a potential tool for studying the cytotoxic mechanisms of Hg2+.


Enthalpy-entropy compensation is not a general feature of weak association.
Relationships between the enthalpy and entropy changes resulting from perturbations of a system have been discussed in the literature for some time. Both positive correlations (compensation) and negative correlations (anti-compensation) between deltaH and DeltaS have been observed in various experimental contexts, including chemical reaction, physical association, solvation, and protein folding. Many examples have been demonstrated to be statistical artifacts, but some are genuine signatures of the perturbations in molecular characteristics. In particular, recent literature claims that compensation is a general feature of bimolecular associations arising from weak intermolecular interactions. We employ a statistical mechanical framework to predict the magnitude and direction of enthalpy-entropy correlation in bimolecular association. The theory links the macroscale thermodynamic correlation to the relationship between the intermolecular potential parameters. Using a harmonic approximation to the Lennard-Jones model and potential parameters taken from the literature, we show examples of both compensation and anti-compensation for gas-phase self-association among five homologous series. Furthermore, an aggregate presentation of data for 48 different chemical species shows no correlation in either direction, for the case of self-association in a dilute gas phase.


Photothermally enhanced photodynamic therapy delivered by nano-graphene oxide.
Graphene with unique physical and chemical properties has shown various potential applications in biomedicine. In this work, a photosensitizer molecule, Chlorin e6 (Ce6), is loaded on polyethylene glycol (PEG)-functionalized graphene oxide (GO) via supramolecular π-π stacking. The obtained GO-PEG-Ce6 complex shows excellent water solubility and is able to generate cytotoxic singlet oxygen under light excitation for photodynamic therapy (PDT). Owing to the significantly enhanced intracellular trafficking of photosensitizers, our GO-PEG-Ce6 complex offers a remarkably improved cancer cell photodynamic destruction effect compared to free Ce6. More importantly, we show that the photothermal effect of graphene can be utilized to promote the delivery of Ce6 molecules by mild local heating when exposed to a near-infrared laser at a low power density, further enhancing the PDT efficacy against cancer cells. Our work highlights the promise of using graphene for potential multifunctional cancer therapies.


Rett syndrome: randomized controlled trial of L-carnitine.
Rett syndrome is a severe neurodevelopmental disorder of unknown etiology, occurring almost exclusively in female patients. The etiology and functional significance of plasma carnitine deficiency seen in some patients with Rett syndrome is unknown. To investigate whether L-carnitine might be of benefit in Rett syndrome, a randomized, placebo-controlled, double-blind crossover trial of L-carnitine has been completed in 35 subjects. Eight-week treatment phases were completed for both a placebo and L-carnitine. Outcome was measured by parents/caregivers and at medical follow-up using three established tools: the Rett Syndrome Motor Behavioral Assessment, the Hand Apraxia Scale, and the Patient Well-Being Index. Analysis comparing change between baseline and week 8 of treatment for L-carnitine and the placebo showed that both parents/caregivers and medical follow-up detected improvements in the subjects' well-being. In addition, medical review showed an improvement on the Hand Apraxia Scale for a higher proportion of girls on L-carnitine. Identification of predictors of clinical improvement has been limited by the power of the study. These findings suggest that L-carnitine is of benefit in some patients with Rett syndrome. While L-carnitine did not lead to major functional changes in ability, the type of changes reported could still have a substantial impact on the girls and their families. Information is still needed, however, to determine if only subgroups of girls with the disorder are responsive to L-carnitine and the appropriate duration of therapy.


[Assessment of standard parenteral nutrition in children].
Nowadays, there is a stronger consensus on the proceedings of nutritional support with parenteral nutrition (PN) in paediatrics, the prescription standards, its formulation, elaboration, and nutritional requirements in order to improve the process quality and the patient's safety. The use of standardized PN solutions in children is rare due to the difficulty to adapt them to every pathophysiologic condition. In order to do so, in 2006 we designed and validated a big range of standard solutions for children weighing more than 10 kg or being older than 1 year. To assess the use of standard PN solutions and their suitability in children from January of 2006 until June of 2008: the frequency of prescription of standard solutions by age, weight, and indication, as well as their modifications. We compared the nutrients given by individualized PN solutions versus the recommendations of the Reference Guidelines and standardised PN. 47 children with a mean weight of 26.6 kg (9-50) and mean age 6.8 years (1-14) received 539 units of PN. Standardized PN (437) were used in 83% of the patients. Their total energy requirements were reached within 1-3 days by using one to three types of formulas. Only 4% (22) of them were modified, with easily feasible changes: volume increase (16), glucose lowering (3), and potassium increase (3). The analysis of the individualized PN in 8 children shows the same trend, with a caloric intake lower than 33% of the recommended one. Standardized PN meet the nutritional requirements in most of the patients according to their morbid condition, highlighting their adaptability and versatility. Their use has eased the prescription-validation-preparation circuit and has improved the efficiency of the process.


ProteinsPlus: a web portal for structure analysis of macromolecules.
With currently more than 126 000 publicly available structures and an increasing growth rate, the Protein Data Bank constitutes a rich data source for structure-driven research in fields like drug discovery, crop science and biotechnology in general. Typical workflows in these areas involve manifold computational tools for the analysis and prediction of molecular functions. Here, we present the ProteinsPlus web server that offers a unified easy-to-use interface to a broad range of tools for the early phase of structure-based molecular modeling. This includes solutions for commonly required pre-processing tasks like structure quality assessment (EDIA), hydrogen placement (Protoss) and the search for alternative conformations (SIENA). Beyond that, it also addresses frequent problems as the generation of 2D-interaction diagrams (PoseView), protein-protein interface classification (HyPPI) as well as automatic pocket detection and druggablity assessment (DoGSiteScorer). The unified ProteinsPlus interface covering all featured approaches provides various facilities for intuitive input and result visualization, case-specific parameterization and download options for further processing. Moreover, its generalized workflow allows the user a quick familiarization with the different tools. ProteinsPlus also stores the calculated results temporarily for future request and thus facilitates convenient result communication and re-access. The server is freely available at http://proteins.plus.


Transforming growth factor beta-1 (TGF-beta 1) released by an Epstein-Barr virus (EBV) positive spontaneous lymphoblastoid cell line from a patient with Kostmann's congenital neutropenia inhibits the growth of normal committed haemopoietic progenitors in vitro.
This study reports the characterization of a spontaneous lymphoblastoid cell line (LCL) raised from the peripheral blood of a patient with Kostmann's congenital neutropenia. The LCL was composed of EBV-infected polyclonal B cells and displayed surface markers and pattern of growth in vitro typical of normal LCLs. The supernatant of the LCL contained a colony inhibiting activity (CIA) that decreased the cloning efficiency of normal committed haemopoietic progenitors and was identified as immunoreactive transforming growth factor beta 1 (TGF-beta 1) by neutralization experiments with a specific antiserum. Control studies with a panel of LCLs spontaneously derived from the peripheral blood of patients seropositive for Epstein-Barr virus (EBV) infections showed that 5/30 LCLs produced a CIA. This CIA was not identifiable as TGF-beta 1 but rather was due to the combined effects of tumour necrosis factor alpha (TNF alpha), tumour necrosis factor beta (TNF beta) and interferon alpha (IFN alpha), that were present in the LCL supernatants. The hypothesis that the B cells latently infected by EBV in vivo and possibly expanded as a consequence of the infection may have contributed to the inhibition of the patient granulopoiesis by releasing TGF-beta 1 will be discussed.


Alteration of CFTR transmembrane span integration by disease-causing mutations.
Many missense mutations in the cystic fibrosis transmembrane conductance regulator protein (CFTR) result in its misfolding, endoplasmic reticulum (ER) accumulation, and, thus, cystic fibrosis. A number of these mutations are located in the predicted CFTR transmembrane (TM) spans and have been projected to alter span integration. However, the boundaries of the spans have not been precisely defined experimentally. In this study, the ER luminal integration profiles of TM1 and TM2 were determined using the ER glycosylation machinery, and the effects of the CF-causing mutations G85E and G91R thereon were assessed. The mutations either destabilize the integrated conformation or alter the TM1 ER integration profile. G85E misfolding is based in TM1 destabilization by glutamic acid and loss of glycine and correlates with the temperature-insensitive ER accumulation of immature full-length CFTR harboring the mutation. By contrast, temperature-dependent misfolding owing to the G91R mutation depends on the introduction of the basic side chain rather than the loss of the glycine. This work demonstrates that CF-causing mutations predicted to have similar effects on CFTR structure actually result in disparate molecular perturbations that underlie ER accumulation and the pathology of CF.


An Extended Swedish National Adoption Study of Bipolar Disorder Illness and Cross-Generational Familial Association With Schizophrenia and Major Depression.
Information about how risk for bipolar disorder is transmitted across generations and how parental risk for bipolar disorder relates to their children's risk for schizophrenia and major depression is limited. To evaluate the sources of parent-offspring transmission of bipolar disorder and its familial cross-generational association with schizophrenia and major depression. Parents and offspring (born 1960-1990) from 4 family types were ascertained from Swedish national samples: intact (offspring, n = 2 175 259), not-lived-with biological father (n = 152 436), lived-with stepfather (n = 73 785), and adoptive (n = 15 624). Data analysis was conducted from October 28, 2019, to January 8, 2020. Three sources of parent-offspring resemblance: genes plus rearing, genes only, and rearing only. Diagnosis of bipolar disorder, broad schizophrenia (ie, schizophrenia as a 3-level variable: unaffected, nonaffective psychosis, and schizophrenia) and major depression obtained from Swedish national registries. Parent-offspring resemblance was assessed primarily by tetrachoric correlation (ie, correlation of liability) and for key results, odds ratios (ORs) from logistic regression. Cross-generational associations of bipolar disorder with broad schizophrenia and major depression were assessed by their transmission from bipolar disorder in parents and transmission to bipolar disorder in offspring. The study population included 2 417 104 individuals of 4 family types (51.8% male and 48.2% female; median age, 41 [range, 25-60] years). For bipolar disorder to bipolar disorder transmission, tetrachoric correlations for 3 types of parent-offspring relationships were statistically homogeneous across family type and mothers and fathers for genes plus rearing (0.25; 95% CI, 0.24-0.26), genes only (0.22; 95% CI, 0.18-0.26), and rearing only (0.07; 95% CI, -0.01 to 0.15). Parallel ORs were 5.20 (95% CI, 4.91-5.50), 3.66 (95% CI, 2.97-4.51), and 1.63 (95% CI, 0.96-2.78). Best-estimate, cross-disorder tetrachoric correlations for 3 types of parent-offspring relationships for bipolar disorder and broad schizophrenia were 0.12 (95% CI, 0.11-0.13) for genes plus rearing, 0.12 (95% CI, 0.09-0.14) for genes only, and -0.03 (95% CI, -0.11 to 0.04) for rearing only, with parallel ORs of 1.95 (95% CI, 1.93-1.97), 2.04 (95% CI, 1.75-2.38), and 0.76 (95% CI, 0.43-1.35). For bipolar disorder and major depression, the parallel tetrachoric correlations were 0.09 (95% CI, 0.07-0.10) for genes plus rearing, 0.04 (95% CI, 0.01-0.07) for genes only, and 0.05 (95% CI, 0.01-0.08) for rearing only; parallel ORs were 1.53 (95% CI, 1.50-1.57), 1.23 (95% CI, 1.13-1.34), and 1.25 (95% CI, 1.09-1.42). Heritability for bipolar disorder was estimated at 0.44 (95% CI, 0.36-0.48). Genetic correlations were estimated at 0.572 (95% CI, 0.560-0.589) between bipolar disorder and broad schizophrenia and 0.302 (95% CI, 0.001-0.523) between bipolar disorder and major depression. The findings of this study suggest that genes are largely responsible for bipolar disorder transmission across generations, although modest rearing effects are also likely present. Cross-generational transmission between bipolar disorder and broad schizophrenia appears to be entirely genetic with a moderate genetic correlation; for bipolar disorder and major depression, transmission appears to result equally from genes and rearing with a modest genetic correlation.


Population heterogeneity in vaccine coverage impacts epidemic thresholds and bifurcation dynamics.
Population heterogeneity, especially in individuals' contact networks, plays an important role in transmission dynamics of infectious diseases. For vaccine-preventable diseases, outstanding issues like vaccine hesitancy and availability of vaccines further lead to nonuniform coverage among groups, not to mention the efficacy of vaccines and the mixing pattern varying from one group to another. As the ongoing COVID-19 pandemic transitions to endemicity, it is of interest and significance to understand the impact of aforementioned population heterogeneity on the emergence and persistence of epidemics. Here we analyze epidemic thresholds and characterize bifurcation dynamics by accounting for heterogeneity caused by group-dependent characteristics, including vaccination rate and efficacy as well as disease transmissibility. Our analysis shows that increases in the difference in vaccination coverage among groups can render multiple equilibria of disease burden to exist even if the overall basic reproductive ratio is below one (also known as backward bifurcation). The presence of other heterogeneity factors such as differences in vaccine efficacy, transmission, mixing pattern, and group size can each exhibit subtle impacts on bifurcation. We find that heterogeneity in vaccine efficacy can undermine the condition for backward bifurcations whereas homophily tends to aggravate disease endemicity. Our results have practical implications for improving public health efforts by addressing the role of population heterogeneity in the spread and control of diseases.