Impact of subsectors of agriculture and economic growth on CO2 emissions in Pakistan: evidence from Environmental Kuznets Curve.
Modernization produces carbon dioxide (CO2) emissions but is also able to achieve sustainable agriculture growth by introducing the concept of renewable energy into the agriculture sector, and through this process reduce the CO2 emissions in the country. The main objective of this research is to check the environmental Kuznets curve hypothesis with CO2 emissions and economic development in renewable energy and agriculture subsectors such as fisheries and crop production in Pakistan. This study covers the time period 1984 to 2020. We have applied the autoregressive distribution lag (ARDL) bound test. The results indicate the existence of long-term association among all variables in the model. The result of co-integration in the short run shows a negative relationship between CO2 emissions and crop production in the current time period, and it shows a positive correlation with the first lag of CO2 emission, which means that current crop production reduces the CO2 emissions by 32% during a year, while in the lag period, it will increase in the short run. In the long run, a 1% increase in crop production will reduce the CO2 emission by 86%. Renewable energy shows a negative relation with CO2 emissions in the short run; a 1% increase in renewable energy will reduced the CO2 by 0.017%. Our results support the existence of the EKC hypothesis for Pakistan. In light of the findings, it is suggested that policy makers should focus more on renewable energy to decrease the level of CO2 as much as possible. Moreover, the government must provide subsidies for machines used for cropping and give special attention to subsectors such as livestock and fisheries.


Preoperative Staphylococcus aureus Screening/Decolonization Protocol Before Total Joint Arthroplasty-Results of a Small Prospective Randomized Trial.
To study the prevalence of Staphylococcus aureus carriage and the impact of preoperatively treating carriers in prosthetic joint infection (PJI), a prospective randomized trial was organized. From January 2010 to December 2012, 1028 of 1305 total joint arthroplasties performed were screened, and selected carriers underwent preoperative decolonization. We observed a 22.2% (228/1028) S aureus colonization rate and only 0.8% methicillin-resistant S aureus. Prosthetic joint infection rate was higher, albeit not significantly, in S aureus carriers than among noncarriers-3.9% (9/228) vs 2.0% (16/800). Treated and untreated carriers showed no significant differences-3.4% (3/89) vs 4.3% (6/139). Most of the 14 S aureus PJI occurred in noncarriers suggesting a lack of causal relation between nasal and PJI S aureus. No clear benefit in screening/decolonizing carriers before total joint arthroplasty could be demonstrated.


Synthesis of trans N-Substituted Pyrrolidine Derivatives Bearing 1,2,4- triazole Ring.
1,2,4-triazoles scaffolds display significant biological activities due to hydrogen bonding, solubility, dipole character, and rigidity. The core motif of 1,2,4-triazoles plays a vital role in clinical drugs such as Rizatriptan (antimigraine), Ribavirin (antiviral), anastrozole (anticancer), etizolam (anxiolytic), estazolam (anticonvulsant), alprazolam (anti-hypnotic), letrozole (aromatase inhibitor), loreclezole (anticonvulsant), trazadone (antidepressant) etc. Methods: Epoxide ring opening of tert-butyl 6-oxa-3-azabicyclo [3.1.0] hexane-3-carboxylate followed by methylation under basic conditions and de-protection gave the corresponding trans 1-(4- methoxypyrrolidin-3-yl)-1H-1,2,4-triazole hydrochloride salt as the precursor. This precursor on reaction with substituted benzoyl chlorides and benzyl bromides gave the desired amide and amine products. A library of 14 N-substituted pyrrolidine derivatives i.e. trans3-methoxy-4-(1H-1,2,4-triazol- 1-yl) pyrrolidin-1-yl) (phenyl)methanone and trans 1-benzyl-4-methoxypyrrolidin-3-yl)-1H-1,2,4- triazole were prepared. Eight novel amides (6a-h) and six amines (8a-f) derivatives were synthesized using 1-(4- methoxypyrrolidin-3-yl)-1H-1,2,4-triazole 4 salt with substituted benzoyl chlorides and benzyl bromides.


Incidence of metal and antibiotic resistance in Pseudomonas spp. from the river water, agricultural soil irrigated with wastewater and groundwater.
A total of 144 isolates of Pseudomonas spp. (48 each from the Yamuna River water, wastewater irrigated soil and groundwater irrigated soil) were tested for their resistance against certain heavy metals and antibiotics. Minimum inhibitory concentrations (MICs) of Hg(2+ ), Cd(2+ ), Cu(2+ ), Zn(2+ ), Ni(2+ ), Pb(2+ ), Cr(3+ ) and Cr(6+ ) for each isolate were also determined. A maximum MIC of 200 μg/ml for mercury and 3,200 μg/ml for other metals were observed. The incidences of metal resistance and MICs of metals for Pseudomonas isolates from the Yamuna water and wastewater irrigated soil were significantly different to those of groundwater irrigated soil. A high level of resistance against tetracycline and polymyxin B (81.2%) was observed in river water isolates. However, 87.5% of Pseudomonas isolates from soil irrigated with wastewater showed resistance to sulphadiazine, whereas 79.1% were resistant to both ampicillin and erythromycin. Isolates from soil irrigated with groundwater exhibited less resistance towards heavy metals and antibiotics as compared to those of river water and wastewater irrigated soil. Majority of the Pseudomonas isolates from water and soil exhibited resistance to multiple metals and antibiotics. Resistance was transferable to recipient Escherichia coli AB2200 strains by conjugation. Plasmids were cured with the curing agent ethidium bromide and acridine orange at sub-MIC concentration.


The use of prostaglandin gel in obstetrics and gynecology.
The use of prostaglandin gel for therapeutic termination of second trimester pregnancy, the management of intrauterine fetal death, ripening of the cervix and induction of labour is reviewed. For these indications, prostaglandin gel is acceptable to patients, effective, and has low incidence of side effects and complications.


Lentivirus vectors in β-thalassemia.
Patients with β-thalassemia major require lifelong transfusions and iron chelation, regardless of the type of causative mutations (e.g., β⁰, β(E)/β⁰). The only available curative therapy is allogeneic hematopoietic transplantation, although most patients do not have an HLA-matched, geno-identical donor, and those who do still risk graft-versus-host disease. Hence, gene therapy by ex vivo transfer of a functional β-globin gene is an attractive novel therapeutic modality. In β-thalassemia, transfer of a therapeutic globin gene does not confer a selective advantage to transduced stem cells, and complex DNA regulatory sequences have to be present within the transfer vector for proper expression. This is why lentiviral vectors have proven especially suited for this application, and the first Phase I/II human clinical trial was initiated. Here, we report on the first gene therapy patient with severe β(E)/β⁰-thalassemia, who has become transfusion-independent, and provide methods and protocols used in the context of this clinical trial.


Women's quality of life is decreased by acute cystitis and antibiotic adverse effects associated with treatment.
Although acute cystitis is a common infection in women, the impact of this infection and its treatment on women's quality of life (QOL) has not been previously described. To evaluate QOL in women treated for acute cystitis, and describe the relationship between QOL, clinical outcome and adverse events of each of the interventions used in the study. Randomized, open-label, multicenter, treatment study. Two family medicine outpatient clinics in Iowa. One-hundred-fifty-seven women with clinical signs and symptoms of acute uncomplicated cystitis. Fifty-two patients received trimethoprim/sulfamethoxazole 1 double-strength tablet twice daily for 3 days, 54 patients received ciprofloxacin 250 mg twice daily for 3 days and 51 patients received nitrofurantoin 100 mg twice daily for 7 days. QOL was assessed at the time of enrollment and at 3, 7, 14 and 28 days after the initial visit. QOL was measured using a modified Quality of Well-Being scale, a validated, multi-attribute health scale. Clinical outcome was assessed by telephone interview on days 3, 7, 14 and 28 using a standardized questionnaire to assess resolution of symptoms, compliance with the prescribed regimen, and occurrence of adverse events. Patients experiencing a clinical cure had significantly better QOL at days 3 (p = 0.03), 7 (p < 0.001), and 14 (p = 0.02) compared to patients who failed treatment. While there was no difference in QOL by treatment assignment, patients experiencing an adverse event had lower QOL throughout the study period. Patients treated with ciprofloxacin appeared to experience adverse events at a higher rate (62%) compared to those treated with TMP/SMX (45%) and nitrofurantoin (49%), however the difference was not statistically significant (p = 0.2). Patients experiencing cystitis have an increase in their QOL with treatment. Those experiencing clinical cure have greater improvement in QOL compared to patients fail therapy. While QOL is improved by treatment, those reporting adverse events have lower overall QOL compared to those who do not experience adverse events. This study is important in that it suggests that both cystitis and antibiotic treatment can affect QOL in a measurable way.


Design of an In Situ Cross-Linked Eutectic Tablet for Enhanced Delivery of Gastro-Sensitive Proteins and Peptides.
In the present study, a eutectic platform was designed as an in situ cross-linked eutectic tablet for structural protection, enhanced intestinal permeation, and controlled release of proteins after oral administration. Physicochemical and physicomechanical analyses of the eutectic tablets were undertaken to elucidate the in situ cross-linking mechanism, thermal transitions, crystallinity, ex vivo permeation, and in vitro release of the protein. Following thermal characterization, results revealed successful eutectic formation with a melting point to 37°C. Protein release from the formulation was controlled over 24 h with a maximum fractional release of ±0.8 for all formulations. The release pattern alternated between phases of burst and slow release which was attributed to the combined effects of swelling, surface erosion, and in situ cross-linking. Mathematical modeling of the protein release kinetics corresponded best with the Higuichi model with near zero-order (R(2) ≈ 0.9787) release. The permeation-enhancing effect of menthol contained within the eutectic powder blend was investigated and results showed an enhanced protein flux (0.0576-0.0720 mg·cm(-2) h(-1)) across the intestinal tissue model compared with a control formulation. Extensive in vitro characterization highlighted the successful design of the eutectic tablets as a potential oral delivery system for proteins with structural protection, enhanced intestinal permeation, and controlled release.


Immunogenicity of subviral herpes simplex virus preparations. I. Formation of neutralizing antibodies in different animal species after administration of herpes simplex virus solubilized antigens.
Production of neutralizing antibodies was followed in guinea pigs, rabbits, hamsters and mice immunized with crude antigen extracts (AM) from human diploid cells infected with herpes simplex virus type 1. The AM induced relatively high levels of neutralizing antibodies in all four species. The antibodies were predominantly complement-requiring and remained so even after administration of repeated AM doses. With the strains used, the antibody response was predominantly type specific and, surprisingly, the type specificity of sera usually increased after administration of repeated doses of AM. Guinea pigs seemed to be the best responsive animal species. They developed the highest levels of antibodies and complement-nonrequiring antibodies were seen in them earlier than in the other animal species. The dose-response experiments carried out in guinea pigs indicated that after a single dose administration the ratio between complement-requiring and complement-nonrequiring antibodies was dependent on the amount of antigen administered. When AM was given without adjuvant less efficient antibody production wws observed than after the administration of the same amount of antigen with adjuvant.


Third International Consensus Conference on lesions of uncertain malignant potential in the breast (B3 lesions).
The heterogeneous group of B3 lesions in the breast harbors lesions with different malignant potential and progression risk. As several studies about B3 lesions have been published since the last Consensus in 2018, the 3rd International Consensus Conference discussed the six most relevant B3 lesions (atypical ductal hyperplasia (ADH), flat epithelial atypia (FEA), classical lobular neoplasia (LN), radial scar (RS), papillary lesions (PL) without atypia, and phyllodes tumors (PT)) and made recommendations for diagnostic and therapeutic approaches. Following a presentation of current data of each B3 lesion, the international and interdisciplinary panel of 33 specialists and key opinion leaders voted on the recommendations for further management after core-needle biopsy (CNB) and vacuum-assisted biopsy (VAB). In case of B3 lesion diagnosis on CNB, OE was recommended in ADH and PT, whereas in the other B3 lesions, vacuum-assisted excision was considered an equivalent alternative to OE. In ADH, most panelists (76%) recommended an open excision (OE) after diagnosis on VAB, whereas observation after a complete VAB-removal on imaging was accepted by 34%. In LN, the majority of the panel (90%) preferred observation following complete VAB-removal. Results were similar in RS (82%), PL (100%), and FEA (100%). In benign PT, a slim majority (55%) also recommended an observation after a complete VAB-removal. VAB with subsequent active surveillance can replace an open surgical intervention for most B3 lesions (RS, FEA, PL, PT, and LN). Compared to previous recommendations, there is an increasing trend to a de-escalating strategy in classical LN. Due to the higher risk of upgrade into malignancy, OE remains the preferred approach after the diagnosis of ADH.


Associations between HOTAIR polymorphisms rs4759314, rs920778, rs1899663, and rs7958904 and risk of primary ovarian insufficiency in Korean women.
We investigated the association between the Hox transcript antisense RNA (HOTAIR) polymorphisms rs4759314, rs920778, rs1899663, and rs7958904 and primary ovarian insufficiency (POI) in Korean women. We conducted a case-control study of 134 Korean women with POI and 383 control individuals with at least one live birth and no history of pregnancy loss. The GT genotype of rs1899663 was associated with a decreased risk of POI compared with other genotypes at that locus. In addition, compared with the wild-type homozygous genotypes, the combination of the AA genotype of rs4759314 and the GC genotype of rs7958904 was associated with a decreased risk of POI (P < 0.05), whereas the combination of the GG genotype of rs1899663 and the GC genotype of rs7958904 was associated with an increased risk of POI (P = 0.003). Haplotype analysis revealed that certain haplotypes involving some or all of the polymorphisms were associated with a decreased risk of POI, whereas other haplotypes were associated with an increased risk of POI. Serum levels of luteinizing hormone, follicle-stimulating hormone, and estradiol differed between patients with POI and control individuals (P < 0.05). Our results suggest that the HOTAIR polymorphisms rs4759314, rs920778, rs1899663, and rs7958904 are involved in POI.


Minimum income protection and European integration: trends and levels of minimum benefits in comparative perspective, 1990-2005.
This article draws attention to the Europeanization of social policy and the development of minimum income protection in a large number of welfare democracies. The empirical analyses are based on unique institutional and comparative data on benefit levels from the Social Assistance and Minimum Income Protection Interim Dataset. There is some evidence of convergence in benefit levels among the European countries in the new millennium, but there is no clear proof of universal ambitions to fight poverty or of the existence of a single European social model. There are still welfare front-runners and those who lag behind in this regard, not only among industrial welfare democracies in general but also in Europe.


Biomarkers for Heart Failure in Asia.
Contributions from the Asian biomedical community to knowledge of biomarkers in heart failure have grown rapidly since 2000. Japan has made world-leading contributions in the discovery and application of cardiac natriuretic peptides as biomarkers in heart failure, but there has been rapid growth in reports from China. Contributions also come from Taiwan, South Korea, Singapore, and Hong Kong. Centers in Asia have established clinical cohorts providing powerful platforms for the discovery and validation of biomarkers in heart failure. This century, Asian enquiry into biomarkers in heart failure will include peptides, cytokines, metabolites, nucleic acids, and other analytes.


Use of magnetic circular dichroism to study dinuclear metallohydrolases and the corresponding biomimetics.
Magnetic circular dichroism (MCD) is a convenient technique for providing structural and mechanistic insight into enzymatic systems in solution. The focus of this review is on aspects of geometric and electronic structure that can be determined by MCD, and how this method can further our understanding of enzymatic mechanisms. Dinuclear Co(II) systems that catalyse hydrolytic reactions were selected to illustrate the approach. These systems all contain active sites with similar structures consisting of two Co(II) ions bridged by one or two carboxylates and a water or hydroxide. In most of these active sites one Co(II) is five-coordinate and one is six-coordinate, with differing binding affinities. It is shown how MCD can be used to determine which binding site--five or six-coordinate--has the greater affinity. Importantly, zero-field-splitting data and magnetic exchange coupling constants may be determined from the temperature and field dependence of MCD data. The relevance of these data to the function of the enzymatic systems is discussed.


Uremic pulmonary edema.
Pulmonary edema fluid analyses and hemodynamic evaluations were performed in two uremic patients with acute pulmonary edema. The colloid osmotic pressure of the pulmonary edema fluid ranged from 57 per cent to 93 per cent that of the serum. Although cardiac function was normal in both patients, the serum colloid osmotic pressure--pulmonary artery wedge pressure gradients were markedly reduced. Uremic pulmonary edema is the result of alterations of pulmonary intravascular Starling forces and increases in pulmonary capillary membrane permeability, allowing for the efflux of protein-rich fluid from the capillaries into the lung.


Low hemoglobin levels and recurrent falls in U.S. men and women: prospective findings from the REasons for Geographic And Racial Differences in Stroke (REGARDS) cohort.
There are few data available on low hemoglobin and incident falls in the general U.S. population. Of 30,239 black and white U.S. adults ≥45 years in the population-based REasons for Geographic And Racial Differences in Stroke study, 16,782 had hemoglobin measured at baseline and follow-up data on falls. Hemoglobin was categorized by 1.0 g/dL increments relative to the World Health Organization anemia threshold (<13.0 g/dL for men, <12.0 g/dL for women). Recurrent falls (≥2 falls in the 6 months after baseline) were assessed during a telephone interview. Recurrent falls occurred in 3.9% of men and 4.8% of women. Compared with those with a hemoglobin level 1 to 2 g/dL above the anemia cut-off, multivariable adjusted odds ratios (95% confidence intervals) for recurrent falls associated with hemoglobin levels ≥3, 2 to <3 and 0 to 1 g/dL above the cut-off point, and 0 to <1 and ≥1 g/dL below the cut-off point were 0.73 (0.45-1.19), 0.84 (0.57-1.24), 1.29 (0.88-1.90), 1.32 (0.0.80-1.2.18) and 2.12 (1.23-3.63), respectively, among men (linear trend P < 0.001), and 1.59 (1.10-2.3), 1.24 (0.95-1.62), 1.42(1.11-1.81), 1.28 (0.91-1.80) and 1.76 (1.13-2.74), respectively, among women (linear trend P = 0.45; quadratic trend P = 0.016). Among men, lower hemoglobin levels were associated with an increased risk for recurrent falls. Although our findings suggest an increased risk for recurrent falls at both lower and higher hemoglobin levels among women, these findings should be confirmed in subsequent studies.


Addition of cisplatin and bleomycin to vincristine-carmustine-doxorubicin-prednisone (VBAP) combination in the treatment of relapsing or resistant multiple myeloma: a Southwest Oncology Group study.
Fifty-five patients with myeloma who had relapsed on or were resistant to melphalan and/or cyclophosphamide with prednisone received vincristine, carmustine, doxorubicin, and prednisone (VBAP) plus cisplatin and bleomycin at 21-day intervals. Eighteen (32.7%) patients responded. The response rate was 38.5% (15 responses among 39 patients) in relapsing patients. Three (18.8%) of 16 patients with resistant myeloma responded. Granulocytopenia was the most frequent toxic effect, and was severe in 12 (22%) patients. Severe thrombocytopenia occurred in seven (13%) patients and severe nausea and vomiting occurred in eight (15%). One patient with previously normal renal function developed renal failure on this regimen. The median survival (100 weeks) in those patients responding to treatment was significantly longer than that in patients not responding (25 weeks; P = 0.001). VBAP plus cisplatin and bleomycin was at least as effective as VBAP but had greater toxicity, expense, and inconvenience; it therefore is not preferable to VBAP.


Major histocompatibility complex and central nervous system involvement by paracoccidioidomycosis.
Paracoccidioidomycosis (PCM) is a chronic granulomatous infectious disease, whose etiologic agent is the fungus Paracoccidioides brasiliensis. The central nervous system (CNS) involvement with paracoccidioidomycosis (NPCM) occurs more frequently than has been admitted in the past. There are some major histocompatibility complex antigen association studies with systemic paracoccidioidomycosis. Some indicate a positive association with HLA antigens, but there is no study with the involvement of the CNS. To investigate why not all cases of systemic PCM show the involvement of the CNS and whether genetic factors are involved, we studied 6 patients with NPCM, from the neuroinfection outpatient clinic. The patients were typed for class I and class II antigens by a microlymphocytoxity standard test. The HLA antigen frequencies found in this study in patients with NPCM were not different from the alleles frequencies observed in the Paraná population.


Identification of denatured enzyme proteins in sodium dodecyl sulfate polyacrylamide gels.
A simple modification of the immunological sandwich method of Muilerman et al. for the identification of denatured enzyme proteins in sodium dodecyl sulfate-polyacrylamide gels is described, enabling the method to be used in principle for any enzyme whose activity is not inhibited by binding to antibodies. An immunological sandwich consisting of denatured enzyme, antibodies, and native enzyme is formed on a nitrocellulose filter blot of the gel, the filter is divided into strips, and each strip is tested for enzyme activity. The presence of enzyme activity serves to identify the region in the gel containing denatured enzyme protein. Experiments with human lysosomal alpha-glucosidase as a model system are described. The method was applied to identify a protein of Mr 125,000 as the main component with UDPgalactose pyrophosphatase activity in a partially purified preparation of the enzyme from rat liver.


Effect of smoking on blood lead levels in workers and role of reactive oxygen species in lead-induced sperm chromatin DNA damage.
To investigate whether cigarette smoking affects the blood lead levels (BLL) and whether exposure to lead introduces sperm chromatin DNA damage in factory workers. Cross-sectional study. A battery plant in Taiwan. Eighty male workers employed within a battery plant. Standard semen analysis was performed according to the World Health Organization guidelines. Assessment of BLL, sperm chromatin DNA structure, reactive oxygen species generation and other conventional parameters of semen quality. As compared with nonsmoking workers, the BLL were found to be considerably higher among smokers. Statistically significant differences were found in the sperm DNA denaturation (alphaT) induction and the percentage of sperm with increased DNA denaturation (COMP alphaT) in workers with moderate BLL (>or=25 microg/dL). After adjustment for smoking propensity, a positive correlation was discernible between BLL and alphaT, COMP alphaT, and morphologic abnormality. Furthermore, alphaT and COMP alphaT were also found to have positive correlations with sperm superoxide anion production. Workers with higher BLL were found to be at a higher risk of sperm morphologic abnormality and chromatin DNA integrity. These data are significant because they can facilitate the estimation of lead exposure in reproductive toxicology.


Insulin absorption in renal proximal tubules: a quantitative immunocytochemical study.
The purposes of the present study are mainly biological concerning proximal tubular handling of insulin: we will study the intracellular transport to subcellular compartments involved in insulin degradation, the specificity and saturability of the luminal endocytic absorption of insulin, the visualization of transtubular transport, and finally, if possible, the evaluation of the relative distribution (accumulation) of insulin in endocytic vacuoles and lysosomes. The second part is methodological: application of quantitative immunocytochemistry to endocytosis, quantitation of the effect of particle size and antigen density on labeling density on tissue sections, labeling at very low antigen densities, and effect of fish gelatin on background. Isolated renal proximal tubules were perfused with native insulin, 125I-insulin, or [leucineB-25]-insulin (2% receptor-binding ability and full immunoreactivity) or exposed to native insulin from the basolateral membranes. In conclusion, the luminal uptake of insulin is of low specificity, as native and [leucineB-25]-insulin were accumulated to the same extent. Endocytic uptake is of high capacity and the mechanism is saturable. Insulin accumulated in endocytic vacuoles and lysosomes, thus following the classical degradation pathway. No other subcellular compartment is associated with insulin degradation. It was not possible to detect the basolateral uptake, indicating loss of immunoreactivity after binding to its receptor. Absolute quantitative immunocytochemistry is applicable in studying endocytosis. The labeling density increases nonproportionally with antigen density probably caused by steric hindrances. Reduction of the particle size (16 to 6 nm) increased the labeling density 17.6 times.


Characterization and clinical implications of different malignant transformation patterns in diffuse low-grade gliomas.
Malignant transformation (MT) of low-grade gliomas (LGGs) to a higher-grade variant seems inevitable, yet it remains unclear which LGG patients will progress to grade 3 or even directly to grade 4 after receiving a long course of treatment. To elucidate this, we conducted a retrospective cohort study based on 229 adults with recurrent LGG. Our study aimed to disclose the characteristics of different MT patterns and to build predictive models for patients with LGG. Patients were allocated into group 2-2 (n = 81, 35.4%), group 2-3 (n = 91, 39.7%), and group 2-4 (n = 57, 24.9%), based on their MT patterns. Patients who underwent MT showed lower Karnofsky performance scale (KPS) scores, larger tumor sizes, smaller extents of resection (EOR), higher Ki-67 indices, lower rates of 1p/19q codeletion, but higher rates of subventricular involvement, radiotherapy, chemotherapy, astrocytoma, and post-progression enhancement (PPE) compared with those in group 2-2 (p < 0.01). On multivariate logistic regression, 1p/19q codeletion, Ki-67 index, radiotherapy, EOR, and KPS score were independently associated with MT (p < 0.05). Survival analyses demonstrated that patients in group 2-2 had the longest survival, followed by group 2-3 and then group 2-4 (p < 0.0001). Based on these independent parameters, we constructed a nomogram model that exhibited superior potential (sensitivity: 0.864, specificity: 0.814, and accuracy: 0.843) compared with PPE in early prediction of MT. Combining the factors of 1p/19q codeletion, Ki-67 index, radiotherapy, EOR, and KPS score that were presented at initial diagnosis could precisely forecast the subsequent MT patterns of patients with LGG.


The in vitro effect on arterial wall function of serum from patients with pregnancy-induced hypertension.
The object of this study was to develop an in vitro model to identify the serum factor(s) responsible for the vascular changes that occur in pregnancy-induced hypertension (preeclampsia). Segments of rabbit carotid artery were perfused for 3 hours with nutrient medium to which serum from normal pregnant women or patients with preeclampsia was added. After perfusion, ring segments of the vessels were cut and mounted in muscle chambers for force measurement. Sensitivity of the vessels to constrictor agents and endothelium-mediated relaxation was studied. The results indicate that arteries exposed to serum from preeclamptic patients developed a 2.9-fold increase in sensitivity to angiotensin II, 1.6-fold increased sensitivity to norepinephrine, and no change of sensitivity to serotonin. These effects were not accompanied by changes in isometric force. Endothelium-mediated relaxation was not affected by serum from preeclamptic patients. These findings confirm clinical observations of increased angiotensin II sensitivity in preeclampsia. The system used may facilitate identification of factors that cause the difference between the sera of normal pregnant women and those with preeclampsia.


Ichthyotoxic activities of Ateleia glazioviana Baill. and Thinouia coriacea Britt.
Ethanolic extracts from leaves and roots of Ateleia glazioviana (Leguminosae-Papilionoideae) and the saponin containing fraction from Thinouia coriacea (Sapindaceae) have been studied for ichthyotoxic activity on Phallocerus caudimaculatus and Geophagus brasiliensis. The ethanolic extracts were used in doses ranging from 50 to 100 ppm, based on the dry residue. The saponin fraction was used at doses of 1-10 ppm. The two plant extracts and saponin fractions were found to be ichthyotoxic, justifying their traditional role as an aid in fishing.


Spectral characteristics of heart rate and blood pressure variability in primary aldosteronism.
A disturbance of the autonomic cardiovascular function has been postulated in primary aldosteronism as a possible mechanism for hypertension. Using the method of spectral analysis of heart rate and blood pressure variability, the aim of this study was to assess sympathovagal interactions modulating cardiovascular function and baroreflex control in patients with primary aldosteronism. Seventeen patients (7 with adenomas and 10 with idiopathic hyperaldosteronism) and a control group of 11 essential hypertensives (EH) and 10 normotensive subjects were studied. Continuous finger blood pressure was measured using a Finapres device and heart rate was measured using an electrocardiographic monitor. The studies were conducted in each patient for 20 min in the supine position and 10 min during a passive head-up tilt to 60 degrees. The low frequency and the low frequency-to-high frequency ratio of systolic and diastolic blood pressures were significantly higher both in primary aldosteronism and in EH patients compared with normotensives (P < .01). Impaired response to tilt was found in the heart rate and blood pressure variability in primary aldosteronism and EH patients; the increase in low frequency was smaller than in normotensives. Baroreflex gain (assessed by alpha index) was less in primary aldosteronism and EH patients as compared with normotensives. These findings may suggest that volume expansion or sympathetically mediated central translocation of the blood volume, or both, could stimulate the cardiopulmonary receptors and impair the baroreflex gain. Our findings suggest that an enhanced peripheral vascular responsiveness to a normal sympathetic outflow may be involved in the pathogenesis of hypertension in primary aldosteronism.


Investigation of HDAC8-ligands' intermolecular forces through molecular dynamics simulations: profiling of non-bonding energies to design potential compounds as new anti-cancer agents.
Histone deacetylases are zinc-dependent isoform enzymes and play important role in cellular homeostasis. Among these, HDAC8 is a potential anticancer drug target. To design new inhibitors using protein-ligand energy profiles, an all atom molecular dynamics (MD) simulations were carried out on nine HDAC8-ligand co-crystals (PDBs: 1T64, 1T69, 1T67, 3F07, 1W22, 1VKG, 5FCW, 3SFF and 3SFH). TSN, SHH, B3N, AGE, NHB, CRI, 5YA, 0DI and 1DI are ligands of PDBs, respectively. For these HDAC8-ligands, relative Gibbs binding free energy (ΔGbind) from MM/PBSA method and non-bonding energies (NBE) are in agreement with each other (r2=0.678). Therefore, the NBEs are used to analyze ligands' sub-structures, namely zinc-binding, linker and CAP groups. For linker/CAP regions, this identified carbonyl, amide, and sulfonamide moieties as desirable and alkyl/aryl moieties as electrostatically unfavourable. Using this information, systematically new compounds were designed and subjected to MD simulations. This resulted in seven compounds (NC-I to NC-VII) with encouraging energy profiles (NBE: -76.25 to -127.09 kcal/mol; ΔGbind: -17.21 to -57.42 kcal/mol) in comparison to that of the HDAC8 ligands (NBE: -46.25 to -106.29 kcal/mol; ΔGbind: -14.74 to -49.52 kcal/mol). From these, NC-VI showed best energy profile (NBE = -126.15 kcal/mol; ΔGbind = -57.42 kcal/mol) suggesting its binding affinity and thermodynamic stability. In addition to this, NC-II and NC-III have shown promising NBE and ΔGbind profiles. These may serve as lead molecules for exploration against HDAC8 in cancer therapy. This has provided a basis for designing new compounds with improved NBE and ΔGbind profiles by modifying the unfavourable or not so favourable regions of ligands. [Formula: see text] Communicated by Ramaswamy H. Sarma.


Using NMR spectroscopy to elucidate the role of molecular motions in enzyme function.
Conformational motions play an essential role in enzyme function, often facilitating the formation of enzyme-substrate complexes and/or product release. Although considerable debate remains regarding the role of molecular motions in the conversion of enzymatic substrates to products, numerous examples have found motions to be crucial for optimization of enzyme scaffolds, effective substrate binding, and product dissociation. Conformational fluctuations are often rate-limiting to enzyme catalysis, primarily through product release, with the chemical reaction occurring much more quickly. As a result, the direct involvement of motions at various stages along the enzyme reaction coordinate remains largely unknown and untested. In the following review, we describe the use of solution NMR techniques designed to probe various timescales of molecular motions and detail examples in which motions play a role in propagating catalytic effects from the active site and directly participate in essential aspects of enzyme function.


Is fourth port really required in laparoscopic cholecystectomy?
Since the advent of four-port laparoscopic cholecystectomy, many modifications regarding port number and size have been tried. The feasibility of three-port technique has been found comparable to the conventional four-port laparoscopic cholecystectomy. To assess the feasibility and safety of three-port laparoscopic cholecystectomy in a prospective study. Between March 2007 and March 2009, fifty patients with cholelithiasis aged between 15 and 56 years underwent three-port cholecystectomy in a prospective study in Government medical college, Srinagar. A single surgeon did all the cases and there was no criterion for the patient selection. These were consecutive fifty surgeries done by the surgeon. The outcome was assessed in terms of intra-operative and post-operative parameters. The mean (range) age was 45 (15-56) years and there were thirty-nine females and eleven males in the study. All the procedures were completed successfully without any conversions to open or any major complications; though three patients needed the addition of a fourth port as in conventional laparoscopic cholecystectomy. The mean (range) operative time was 55 (30-90) min and the average blood loss was 30 ml. The mean (range) hospital stay was 1 (1-3) days. All patients returned to routine work within 1 week of surgery. The mean follow-up was 5 (2-7) months. We conclude, from the results above, that three-port laparoscopic cholecystectomy is safe and feasible. There are only two visible surgical scars, better cosmetic appearance with no increased risk of bile duct injury. It reduces the manpower in the form of a second assistant. Thus, it can be recommended as a safe alternative procedure to conventional four-port laparoscopic cholecystectomy.


Investigation of mitochondrial biogenesis defects in single substantia nigra neurons using post-mortem human tissues.
Mitochondrial respiratory chain deficiency and mitochondrial DNA deletions are reported in substantia nigra neurons from healthy aged and Parkinson's disease cases, with extensive neuronal loss only seen in the latter. This study aimed to understand the pathological relevance of mitochondrial defects for neuronal survival. Using post-mortem human midbrain, substantia nigra neurons exposed to different types of mitochondrial defects (including mitochondrial DNA point mutations, single and multiple deletions) were compared to neurons from healthy aged and Parkinson's disease cases (either sex) at a single neuronal level. We identified mitochondrial deficiencies in all cases, though these deficiencies were more severe in the mitochondrial disease patients with multiple deletions. A significant reduction in TFAM expression was detected in Parkinson's disease compared to cases with other mitochondrial defects. Higher mitochondrial DNA copy number was detected in healthy aged neurons, despite a deletion level equivalent to Parkinson's disease. Our data support that in individuals with pathogenic mitochondrial defects, neurons respond to mitochondrial defect to survive and such an adaptation may involve TFAM.


Mechanical analysis of the lumbar vertebrae in a three-dimensional finite element method model in which intradiscal pressure in the nucleus pulposus was used to establish the model.
We established a three-dimensional finite element method (FEM) model of the 4th and 5th vertebrae, using computed tomography (CT) images (2-mm slice thickness) of a healthy 29-year-old man. Because of the lack of specific data regarding the material characteristics of the nucleus pulposus of intervertebral discs, we used intradiscal pressure in the nucleus pulposus to establish the model. We referred to data from Nachemson and from Sato et al. regarding intradiscal pressure and to the methods of Shirazi-Adl for data for other material characteristics (see text for these references). The mid-position model bears a load of 294 N in the vertical direction, while the models of the flexed and extended positions bear loads of 15 N-m. In addition, a degenerative disc model without intradiscal pressure was created for the standing model. The use of these models allowed the investigation of von Mises stress on the vertebral endplates and the annulus fibrosus. We also examined von Mises stress on the facet joint in normal and degenerative disc models. There was increased von Mises stress on the vertebral endplate in the anterior, center portions. von Mises stress on the annulus fibrosus increased in the posterior portion, the entrance to the neural foramen, and the exit of the neural foramen. von Mises stress was greater during flexion in the posterior portion; in particular, increasing to about 1.6 times the level seen with other postures. No changes were observed in von Mises stress on the vertebral endplates or annulus fibrosus in the degenerative disc model, but von Mises stress on the facet joints was about 2.5 times that seen in the normal disc model.


Analysis of competitive interactions in triocultures of Drosophila melanogaster.
Competitive interactions in complex mixtures of genotypes have rarely been studied despite their obvious importance in both natural and commercial populations. Here, we describe a procedure for the analysis of competition in tripartite mixtures of Drosophila melanogaster genotypes. We have utilised a substitution design coupled with a yield-density regression analysis which describes intra- and inter-genotypic competitive effects in terms of simple linear parameters. The experimental design allows any of the competitors to be considered as the primary or indicator genotype and also incorporates variation in the relative proportions of the two associate competitors. The regression parameters are used to derive estimates of the competitive pressure exerted by each associate on the indicator genotype and also the response or sensitivity of the indicator to the competitive pressure which it faces in mixed culture. The results indicate that the joint pressure exerted by the paired associate genotypes in trioculture is equal to the sum of the individual pressures of those associates. This additive relationship holds for a variety of indicator genotypes isolated from the Texas population and appears to be a general property of Drosophila competition. We identified one indicator genotype which consistently departed from this relationship although additivity of joint pressures could be restored in combination with particular associate genotypes. The possible role of larval interference in the determination of these interactions is discussed.


Loss of heterozygosity on the short arm of chromosome 3 in sporadic, von Hippel-Lindau disease-associated, and familial pheochromocytoma.
Pheochromocytomas occur sporadically and are associated with several dominantly inherited cancer syndromes, including von Hippel-Lindau (VHL) disease. We examined 14 pheochromocytomas (four from VHL patients, nine from sporadic patients, and one from a patient with familial pheochromocytoma) for loss of heterozygosity on chromosome arm 3p by using the polymerase chain reaction and restriction fragment length polymorphisms at eight loci. Loss of heterozygosity was detected in 8 of 14 pheochromocytomas examined: in three of the four VHL-associated tumors, in four of the nine sporadic tumors, and in the familial pheochromocytoma-associated tumor. Deletion of the inherited wild-type VHL allele was demonstrated in both informative VHL-associated pheochromocytomas, demonstrating involvement of VHL in pheochromocytoma development. However, because VHL mutations have not been detected in sporadic pheochromocytomas, VHL and/or another chromosome arm 3p gene may be involved in the etiology of these tumors.


Non-invasive quantification of cerebral blood flow for rats by microPET imaging of 15O labelled water: the application of a cardiac time-activity curve for the tracer arterial input function.
In-vivo quantitative cerebral blood flow (CBF) measurement using positron emission tomography (PET) has typically employed invasive arterial blood sampling procedure to determine the arterial input function (AIF). The present study was performed to provide a non-invasive quantitative CBF measurement technique for rats using a dedicated animal PET. CBF was measured in 10 male rats (Fischer 344, 247-290 g) under alpha-chloralose anesthesia (30 mg x kg . h, intravenous infusion) by dynamic PET imaging employing the intravenous bolus injection of H2(15)O. Unlike other conventional PET methods, no arterial blood sampling was employed. Instead, a cardiac time-activity curve (TAC) obtained from the dynamic PET imaging was used to determine the AIF. For the validation of this technique, CBF was also measured by calculating the washout rate of the tracer (H2(15)O) following an intracarotid bolus injection. CBF measurements by two independent methods were done while modulating and maintaining the body temperature at two different levels (32+/-1 and 37+/-1 degrees C by the rectal temperature). Two methods were compared by the linear regression analysis. CBF (ml x 100 g x min) values (mean+/-SD) were 45.2+/-6.05 (intravenous) and 47.4+/-8.64 (intracarotid) at the hypothermic condition (32 degrees C), and 55.1+/-4.88 (intravenous) and 54.4+/-4.60 (intracarotid) at the normothermic condition (37 degrees C). There was a good agreement between the two methods (r=0.70). Our cardiac TAC analysis technique for small animals can be used for the non-invasive quantification of CBF using the PET-based in-vivo imaging technique.


The effectiveness of market orientation in the logistic industry: A focus on SMEs in an emerging country.
Although the interrelationship between market orientation, innovation, and performance has been intensively examined in the literature, there is a lack of understanding of this in the contexts of small and medium enterprises (SMEs) operating in Vietnamese logistic industry. Thus, this study aims to draw upon the resource-based view to examine the mediating effects of incremental and radical innovation on the link between market orientation and organizational performance. Data collected from 153 logistic SMEs in Vietnam were used to assess the proposed model. Partial least square structural equation modelling was used to examine data. The results suggest that incremental and radical innovation partially mediate the link between market orientation and organizational performance. In this regard, this study contributes to logistic literature by showing the antecedents and consequences of innovation. In addition, this study contributes to market orientation literature by assessing the consequences of market orientation in Vietnam. This study also contributes to the refinement of resource-based view. Finally, this study also provides practical implications for managers manage logistic SMEs in Vietnam.


Oxytocin drives prosocial biases in favor of attractive people.
Current perspectives on attractiveness-related prosocial biases emphasize the contribution of evolutionarily shaped mating drives. Here, we extend these concepts by highlighting the pivotal role of the hypothalamic peptide oxytocin in augmenting the salience and rewarding value of social stimuli, including the partner's face, thereby fostering social bonding in general and the stability of monogamous pair bonds and offspring care in particular.


Financial burden on the families of patients with hepatitis B virus-related liver diseases and the role of public health insurance in Yunnan province of China.
To investigate the financial burden of patients who had various stages of hepatitis B virus-related diseases and the level of alleviation from financial burden by health insurance schemes in Yunnan province of China. A cross-sectional survey. Patients' information was consecutively recorded at the First Affiliated Hospital of Kunming Medical University, from December 2012 to June 2013. Consecutive cases of hepatitis B virus (HBV) (520), compensated cirrhosis (91), decompensated cirrhosis (198) and hepatocellular carcinoma (HCC) (131) were recruited from the outpatient and inpatient departments. The total direct costs, hospital charge, outpatient costs, hospitalization fees being reimbursed and household catastrophic health expenditure were estimated for each disease group. The average annual direct costs for each disease group were 19,496 RMB for HBV, 28,466 RMB in compensated cirrhosis, 46,061 RMB for decompensated cirrhosis, and 33,044 RMB for HCC patients. Catastrophic health expenditure occurred in all four groups. Health insurance reimbursement released the financial burden incurred by medical expenses of patients under a high level of household economic status. Public health insurance schemes helped the patients to various extents. Among these patient groups, direct costs represent a significant economic burden. Health expenditure and financing systems must be considered to prevent the increase of household catastrophe, particularly among the poor.


Short- and intermediate-term exposure to ambient fine particulate elements and leukocyte epigenome-wide DNA methylation in older men: the Normative Aging Study.
Several epigenome-wide association studies (EWAS) of ambient particulate matter with aerodynamic diameter ≤ 2.5 µm (PM2.5) have been reported. However, EWAS of PM2.5 elements (PEs), reflecting different emission sources, are very limited. We performed EWAS of short- and intermediate-term exposure to PM2.5 and 13 PEs. We hypothesized that significant changes in DNAm may vary by PM2.5 mass and its elements. We repeatedly collected blood samples in the Normative Aging Study and measured leukocyte DNA methylation (DNAm) with the Illumina HumanMethylation450K BeadChip. We collected daily PM2.5 and 13 PEs at a fixed central site. To estimate the associations between each PE and DNAm at individual cytosine-phosphate-guanine (CpG) sites, we incorporated a distributed-lag (0-27 d) term in the setting of median regression with subject-specific intercept and examined cumulative lag associations. We also accounted for selection bias due to loss to follow-up and mortality prior to enrollment. Significantly differentially methylated probes (DMPs) were identified using Bonferroni correction for multiple testing. We further conducted regional and pathway analyses to identify significantly differentially methylated regions (DMRs) and pathways. We included 695 men with 1,266 visits between 1999 and 2013. The subjects had a mean age of 75 years. The significant DMPs, DMRs, and pathways varied by to PM2.5 total mass and PEs. For example, PM2.5 total mass was associated with 2,717 DMPs and 10,470 DMRs whereas Pb was associated with 3,173 DMPs and 637 DMRs. The identified pathways by PM2.5 mass were mostly involved in mood disorders, neuroplasticity, immunity, and inflammation, whereas the pathways associated with motor vehicles (BC, Cu, Pb, and Zn) were related with cardiovascular disease and cancer (e.g., "PPARs signaling"). PM2.5 and PE were associated with methylation changes at multiple probes and along multiple pathways, in ways that varied by particle components.


[Nuclear magnetic resonance tomography studies of orbital space occupying lesions].
A comparative study of MRI, CT and ultrasound of orbital tumors revealed that only for malignant melanomas and orbital pseudotumors MRI had the most diagnostic relevance. Malignant melanomas could be differentiated from a haematoma due to their characteristic T1 and T2 signal intensity. MRI is impaired by its long examination time and the danger of quality reduction caused by motion artifacts.


Some general regularities of synergistic interaction of hyperthermia with various physical and chemical inactivating agents.
Various cellular systems have been analysed for synergistic interactions of the simultaneous action of hyperthermia with physical agents including ionizing radiation, ultraviolet light, and ultrasound, and with chemical agents including thio-TEPA and cis-DDP. Some general rules of synergistic interaction were revealed. First, for every constant rate or intensity of exposure to physical factors or concentration of chemical agents, synergy can be observed only within a certain temperature range that is different for various cellular systems. Secondly, within this range, there is a specific temperature that maximizes the synergistic effect. Any deviation of temperature from the optimal one results in a reduction in synergy. Thirdly, the rate of exposure to physical agents or the concentration of chemical agents strongly influences the synergy; i.e. as the dose rate or concentration is reduced, the temperature for maximum synergism decreases. Possible interpretations of the mechanisms responsible for these relationships of synergy are discussed.


Synthesis and 5 alpha-reductase inhibitory activities of benzofuran derivatives with a carbamoyl group.
A series of 2-phenylbenzofuran derivatives with a diphenylmethylcarbamoyl group at the 5 or 6 position of the benzofuran ring were synthesized and evaluated for rat and human testosterone 5 alpha-reductase inhibitory activities in vitro. They had inhibitory activities against both enzymes and the 6-carbamoyl derivatives tended to be more potent than the 5-carbamoyl compounds.


Simultaneous quantification of tafetinib (SIM010603), a novel potent inhibitor of receptor tyrosine kinase, and its major metabolite in dog plasma by HPLC-ESI/MS/MS and its application to a pharmacokinetic study.
This study presents a high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (HPLC-ESI/MS/MS) technique for the simultaneous determination of tafetinib (SIM010603) and its main metabolite (M1) in dog plasma by using Prazosin hydrochloric acid as the internal standard (IS). Both compounds were extracted from dog plasma with ethyl acetate and were separated by HPLC on a reversed phase C₁₈ column with a mobile phase of 10 mM ammonium acetate buffer containing 0.1% formic acid-acetonitrile (40:60, v/v) at a flow rate of 0.2 mL/min. For quantification, the triple-quadruple MS was used in selected reaction monitoring (SRM) mode. The monitored transitions were m/z 425.3 → 309.2 for tafetinib, m/z 397.2 → 309.2 for M1 and m/z 384.2 → 247.1 for IS. The developed method had a short run time of 4 min and good linearity was observed over a wide range of 1-1000 ng/mL for the two compounds. The method was successfully applied in the pharmacokinetic study of tafetinib and M1 in dog.


Pharmacokinetics of Single and Multiple Doses of Omega-3 Carboxylic Acids in Healthy Chinese Subjects: A Phase I, Open-Label Study.
In patients with coronary heart disease undergoing primary prevention, hypertriglyceridemia is a residual risk for cardiovascular events. Omega-3 carboxylic acid (OM3-CA), a mixture of the free fatty acid forms of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), may be beneficial in reducing triglyceride levels. As part of the clinical development program of OM3-CA in China, this phase I study evaluated the pharmacokinetics, safety, and tolerability profile of OM3-CA in healthy subjects. The pharmacokinetic results of this study were also compared with those of available data for Western populations. Fourteen healthy Chinese subjects (aged 18-45 years) received once-daily oral OM3-CA 4 g for 14 consecutive days. Pharmacokinetic parameters were assessed from both baseline-uncorrected and baseline-corrected plasma concentrations vs time profile of EPA, DHA, and EPA plus DHA. Following single and multiple oral doses of OM3-CA, the absorption of EPA, DHA, and EPA plus DHA was steady with median tmax occurring at 5.5-6 hours after both single and multiple dosing. Close to steady-state concentrations in plasma were reached after 14 days of continuous once-daily dosing, and accumulation was confirmed for EPA, DHA, and EPA plus DHA. Of the 14 subjects treated with OM3-CA, 6 (42.9%) reported at least 1 adverse event (diarrhea) during the study, which was determined as mild and treatment emergent. No serious adverse events were reported. In summary, the pharmacokinetic profile of oral OM3-CA 4 g after single and multiple dosing in healthy Chinese subjects is consistent with that observed in other ethnic populations.


A gravimetric assessment of the Gotthard Base Tunnel geological model: insights from a novel gravity terrain-adaptation correction and rock physics data.
The Gotthard Base Tunnel (GBT) is a 57 km long railway tunnel, constructed in the Central Alps in Switzerland and extending mainly North-South across numerous geological units. We acquired 80 new gravity data points at the surface along the GBT profile and used 77 gravity measurements in the tunnel to test and constrain the shallow crustal, km-scale geological model established during the tunnel construction. To this end, we developed a novel processing scheme, which computes a fully 3D, density-dependent gravity terrain-adaptation correction (TAC), to consistently compare the gravity observations with the 2D geological model structure; the latter converted into a density model. This approach allowed to explore and quantify candidate rock density distributions along the GBT modelled profile in a computationally-efficient manner, and to test whether a reasonable fit can be found without structural modification of the geological model. The tested density data for the various lithologies were compiled from the SAPHYR rock physical property database. The tested models were evaluated both in terms of misfit between observed and synthetic gravity data, and also in terms of correlation between misfit trend and topography of the target profile. The results indicate that the locally sampled densities provide a better fit to the data for the considered lithologies, rather than density data averaged over a wider set of Alpine rock samples for the same lithology. Furthermore, using one homogeneous and constant density value for all the topographic corrections does not provide an optimal fit to the data, which instead confirms density variations along the profile. Structurally, a satisfactory fit could be found without modifying the 2D geological model, which thus can be considered gravimetry-proof. From a more general perspective, the gravity data processing routines and the density-dependent corrections developed in this case study represent a remarkable potential for further high-resolution gravity investigations of geological structures. The online version contains supplementary material available at 10.1186/s00015-022-00422-z.


Rational design of peptide-based implants for corneal bioengineering.
Regeneration of damaged cornea can save vision for millions of patients. Most of these patients are waiting for transplantation of a donor cornea or suitable substitute to restore vision. Although donor cornea transplantation is the most clinically accepted treatment, shortage of donor cornea results in almost 69 out of every 70 patients untreated with the waiting list for transplantation drastically increasing every year according to a prepandemic estimation. Therefore, corneal replacements are coming up as a cutting-edge alternative strategy. In view of the peptides, especially collagen-like peptides and peptide amphiphiles with bioactive functional motifs demonstrate promising avenue for the corneal tissue engineering and promoting regeneration, by their hierarchical self-assembling propensity to acquire desired nano- to macroscale 3D architecture. Here, we analyze rational peptide designing, self-assembly, and strategies of peptide/peptide-based nanoscale building blocks to create the extracellular matrix mimetic implants for functional regeneration of the cornea.


Cannabinoid receptor blockade reveals parallel plasticity mechanisms in different layers of mouse visual cortex.
The ocular dominance (OD) shift that occurs in visual cortex after brief monocular deprivation (MD) is a classic model of experience-dependent cortical plasticity. It has been suggested that OD plasticity in layer 2/3 of visual cortex precedes and is necessary for plasticity in the thalamocortical input layer 4. Here, we show in mouse visual cortex that rapid OD plasticity occurs simultaneously in layers 2/3 and 4. Remarkably, pharmacological blockade of cannabinoid receptors completely prevents the OD shift in layer 2/3, leaving plasticity intact in layer 4. Thus, experience-dependent cortical modifications in layers 2/3 and 4 can occur in parallel, via distinct mechanisms. These findings simplify the mechanistic description of plasticity in layer 4, force a revision in the interpretation of previous studies in which laminar differences in OD plasticity mechanisms were unrecognized, and have important implications for the therapeutic use of cannabinoid receptor antagonists in humans.


Rescue treatment for noninvasive ventilation failure due to interface intolerance with remifentanil analgosedation: a pilot study.
To assess the feasibility of remifentanil-based sedation in hypoxemic acute respiratory failure (HARF) patients refusing to continue noninvasive ventilation (NPPV) for intolerance to two different interfaces-helmet and total face mask. Prospective uncontrolled clinical investigation in a 14-bed ICU of an university hospital in Italy. Thirty-six patients with persistent severe HARF who complained of discomfort and asked for interruption of NPPV session. Patients started sedation with remifentanil (0.025 μg kg(-1) min(-1)) and the infusion rate was increased by 0.01 μg kg(-1) min(-1) every minute to a maximum of 0.12 μg kg(-1) min(-1) to obtain patient comfort. Twenty-two out of 36 patients (61%) with median (IQR) SAPS II score of 32 (30, 38) continued the NPPV treatment after the introduction of remifentanil infusion. In this success group, median (IQR) respiratory rate decreased from 34 (31, 37) to 24 (20, 26) min(-1) (p < 0.0001) and PaO(2)/FiO(2) ratio increased from 156 (144, 176) to 270 (210, 300) mmHg (p < 0.0001) after 1 h of NPPV with remifentanil-analgosedation either with helmet or total face mask. Fourteen patients failed to continue the noninvasive treatment and were intubated after a mean of 2.5 ± 2.3 h; they showed a respiratory rate decrease from 35 (30, 38) to 27 (25, 35) min(-1) (p = 0.02) and an inability to increase the PaO(2)/FiO(2) ratio above 180 mmHg. The ICU mortality in the failure group patients was 50 versus 14% in the NPPV success group (p < 0.05). The mean remifentanil dose administered was 0.07 ± 0.03 μg kg(-1) min(-1). This clinical study suggests that a remifentanil-based sedation protocol can decrease the rate of failure in patients with intolerance to NPPV.


Retrospective review of clinical presentations, microbiology, and outcomes of patients with psoas abscess.
The clinical features of patients with psoas abscess in a local setting, including the microbiology, incidence and aetiologies, have not been well described. This study aimed to review such clinical features and patient outcomes. Retrospective case series. Regional hospital, Hong Kong. Adults (aged ≥18 years) with psoas abscess admitted to Tuen Mun Hospital from 1 January 2006 to 31 December 2010 were included. The clinical presentations, aetiologies, microbiology, treatments, hospital stays, intensive care unit admissions, and outcomes of these patients were reviewed. The series entailed 42 patients, five of whom developed their psoas abscess after admission and seven were intravenous drug abusers. The most common presenting symptom in community-onset cases was back, hip, or thigh pain (43% [16/37]). Fever was present at presentation in 41% (15/37) of these patients, four of whom presented with fever only. The diagnosis was made by computed tomography in 95% (40/42) of these cases. In all, 23 abscesses were considered secondary; the most common aetiology being infective spondylitis or spondylodiscitis. The commonest causative organism for a primary psoas abscess was methicillin-sensitive Staphylococcus aureus, while for secondary abscesses they were more commonly from the gastro-intestinal and genitourinary tracts. Overall in-hospital mortality rate was 14% (6/42). Secondary psoas abscess patients had longer hospital stays (mean, 62 vs 34 days; P=0.007). Psoas abscess is an uncommon condition. Most patients presented with only non-specific symptoms leading to difficulty in making an early diagnosis. In more than half of these patients, the psoas abscesses were secondary, the aetiology of which differed from reported overseas experience.


Involvement of central GABA receptors in the regulation of blood pressure and heart rate of anesthetized cats.
Intracerebroventricular (i.c.c.) injections of muscimol (0.03-3 microgram/kg) and GABA (30-1000 microgram/kg) caused dose-related reductions in blood pressure and heart rate of anesthetized cats whereas glycine (30-3000 microgram/kg) was much less effective. I.v. administration of muscimol or GABA had little or no effect. Renal sympathetic nerve discharge was also reduced by central GABA receptor stimulation. The effects of both muscimol and GABA were reversed by either picrotoxin or bicuculline administered intravenously. The results suggest an important role for GABA receptors in the central regulation of blood pressure and heart rate.


The emerging global epidemic of paediatric inflammatory bowel disease--causes and consequences.
Two decades ago, paediatric inflammatory bowel disease (IBD) drew only modest interest from the international paediatric community. Since then, dramatically globally increasing incidence rates have made childhood-onset IBD a priority for most paediatric gastroenterologists. The emerging pandemia of paediatric IBD has fuelled a quest to identify the recent changes in early life exposures that could explain the increasing risk for IBD amongst today's children. Treatment of children with IBD should aim for symptom control but should also target restoration of growth and prevention of pubertal delay. The paediatric IBD phenotype seems to be characterized by more extensive disease location, and some comparative studies have suggested that childhood-onset IBD also represents a more severe phenotype than the adult-onset IBD form. In this review, we analyse recent global incidence trends of paediatric IBD. We present an update on the known and suggested risk factors that could explain the emerging global epidemia of paediatric IBD. We also draw attention to differences in treatment between children and adults with IBD. Finally, we highlight latest follow-up studies that question the proposed dynamic and aggressive nature of childhood-onset IBD.


Multiplicity in type V adenylylcyclase: type V-a and type V-b.
Type V mammalian adenylylcyclase cDNA was originally isolated from two animal species, the dog and rat. The amino acid sequences from the two species are highly homologous, but completely different in the putative N-terminal, cytoplasmic region. Northern blot analysis using oligonucleotide probes unique to either of the two clones has revealed that the two forms of type V adenylylcyclase mRNA, canine form (= type V-a) and rat form (= type V-b), are co-expressed as splicing variants in both species. Genomic Southern blot analysis has suggested that the two forms are the products of a single gene. When overexpressed, however, deletion of the N-terminal domain did not alter any biochemical properties. Thus multiple splicing variants with unique N-terminal amino acid sequences of type V adenylylcyclase can be generated from a single gene, however, biochemical properties of these variants may not be different.


Weak anion-exchange mixed-mode materials to selectively extract acidic compounds by stir bar sorptive extraction from environmental waters.
In this study, the first example of a polytetrafluoroethylene (PTFE)-based magnet coated with weak anion exchange (WAX) monolith as novel support for stir bar sorptive extraction (SBSE) is presented. Firstly, the PTFE magnets were properly modified and vinylized in order to immobilize polymer monoliths onto its surface. Then, a glycidyl methacrylate monolith was prepared and modified with ethylenediamine (EDA) to create weak anion exchanger via ring opening reaction of epoxy groups. The prepared covalently immobilized EDA-modified monoliths onto PTFE magnet exhibited good stability and reusability. Application of resulting material as stir bar for SBSE was investigated for a series of acidic compounds that includes acesulfame, saccharin, diclofenac or ibuprofen, among others as target compounds. Firstly, the SBSE conditions were optimized to promote the WAX interactions with the target compounds achieving recoveries from 37 to 75% and enable the selective extraction of these compounds as it provided values of% matrix effect from 17 to -13% when they were determined by SBSE followed by liquid chromatography - tandem mass spectrometry. The analytical methodology, was then validated and applied for the determination of the target solutes in environmental water samples, which were found at concentration up to 2500 ng L-1 in river waters.


The dynamics of molecular, immune and physiological features of the host and the gut microbiome, and their interactions before and after onset of laying in two hen strains.
Aggregation of data, including deep sequencing of mRNA and miRNA data in jejunum mucosa, abundance of immune cells, metabolites, or hormones in blood, composition of microbiota in digesta and duodenal mucosa, and production traits collected along the lifespan, provides a comprehensive picture of lifelong adaptation processes. Here, respective data from two laying hen strains (Lohmann Brown-Classic (LB) and Lohmann LSL-Classic (LSL) collected at 10, 16, 24, 30, and 60 wk of age were analyzed. Data integration revealed strain- and stage-specific biosignatures, including elements indicative of molecular pathways discriminating the strains. Although the strains performed the same, they differed in the activity of immunological and metabolic functions and pathways and showed specific gut-microbiota-interactions in different production periods. The study shows that both strains employ different strategies to acquire and maintain their capabilities under high performance conditions, especially during the transition phase. Furthermore, the study demonstrates the capacity of such integrative analyses to elucidate molecular pathways that reflect functional biodiversity. The bioinformatic reduction of the multidimensional data provides good guidance for further manual review of the data.


GFI1 proteins orchestrate the emergence of haematopoietic stem cells through recruitment of LSD1.
In vertebrates, the first haematopoietic stem cells (HSCs) with multi-lineage and long-term repopulating potential arise in the AGM (aorta-gonad-mesonephros) region. These HSCs are generated from a rare and transient subset of endothelial cells, called haemogenic endothelium (HE), through an endothelial-to-haematopoietic transition (EHT). Here, we establish the absolute requirement of the transcriptional repressors GFI1 and GFI1B (growth factor independence 1 and 1B) in this unique trans-differentiation process. We first demonstrate that Gfi1 expression specifically defines the rare population of HE that generates emerging HSCs. We further establish that in the absence of GFI1 proteins, HSCs and haematopoietic progenitor cells are not produced in the AGM, revealing the critical requirement for GFI1 proteins in intra-embryonic EHT. Finally, we demonstrate that GFI1 proteins recruit the chromatin-modifying protein LSD1, a member of the CoREST repressive complex, to epigenetically silence the endothelial program in HE and allow the emergence of blood cells.


Molecular characterization of eutF mutants of Salmonella typhimurium LT2 identifies eutF lesions as partial-loss-of-function tonB alleles.
The eutF locus of Salmonella typhimurium LT2 was identified as a locus necessary for the utilization of ethanolamine as a sole carbon source. Initial models suggested that EutF was involved in either ethanolamine transport or was a transcriptional regulator of an ethanolamine transporter. Phenotypic characterization of eutF mutants suggested EutF was somehow involved in 1,2-propanediol, propionate, and succinate utilization. Here we provide evidence that two alleles defining the eutF locus, Delta903 and eutF1115, are partial-loss-of-function tonB alleles. Both mutations were complemented by plasmids containing a wild-type allele of the Escherichia coli tonB gene. Immunoblot analysis using TonB monoclonal antibodies detected a TonB fusion protein in strains carrying eutF alleles. Molecular analysis of the Delta903 allele identified a deletion that resulted in the fusion of the 3' end of tonB with the 3' end of trpA. In-frame translation of the tonB-trpA fusion resulted in the final 9 amino acids of TonB being replaced by a 45-amino-acid addition. We isolated a derivative of a strain carrying allele Delta903 that regained the ability to grow on ethanolamine as a carbon and energy source. The molecular characterization of the mutation that corrected the Eut- phenotype caused by allele Delta903 showed that the new mutation was a deletion of two nucleotides at the tonB-trpA fusion site. This deletion resulted in a frameshift that replaced the 45-amino-acid addition with a 5-amino-acid addition. This change resulted in a TonB protein with sufficient activity to restore growth on ethanolamine and eut operon expression to nearly wild-type levels. It was concluded that the observed EutF phenotypes were due to the partial loss of TonB function, which is proposed to result in reduced cobalamin and ferric siderophore transport in an aerobic environment; thus, the eutF locus does not exist.


Prevalence and characteristics of sadistic personality disorder in an outpatient veterans population.
This study is designed to examine the prevalence, demographics, and the Axis I and Axis II diagnoses of sadistic personality disorder (SPD) in a previously unexamined population (outpatient veterans). It also represents the first attempt to examine the family history of those with SPD. Standardized methods were used to select 144 nonpsychotic subjects from a veterans psychiatric outpatient clinic. The population was then divided into three groups: those with SPD, those without SPD but with antisocial personality disorder or traits, and those with no Axis II disorders. A fourth group consisted of 28 control veterans without psychopathology. Subjects with SPD tended to be younger and to have lower scores on the Global Assessment Scale than did clinical control subjects without personality disorders; they also had significantly more bipolar and panic disorder than were found in the antisocial traits group. High levels of depression and alcohol dependence were also present. SPD traits correlated with DSM-III-R dramatic cluster, compulsive, passive-aggressive, and self-defeating personality disorders. The SPD group could be distinguished from other groups on the basis of family history. It was concluded that sadistic personality traits and disorders are prevalent (8.1%), associated with reduced functioning, and may have specific associations with certain Axis I and Axis II disorders. It is possible that they have a distinct familial pattern.


Effect of isoflavone from Flemingia vestita (Fabaceae) on the Ca2+ homeostasis in Raillietina echinobothrida, the cestode of domestic fowl.
The alcoholic crude root-peel extract of Flemingia vestita and its major isoflavone, genistein, have been shown to have a vermifugal/vermicidal effect by causing a flaccid paralysis accompanied by alterations in the structural architecture of the tegumental interface and metabolic activity in Raillietina echinobothrida, the cestode of domestic fowl. In the present study, the crude root-peel extract and pure genistein were tested in vitro with respect to Ca2+ homeostasis and the occurrence of some metal ions was detected in the parasite. Live cestodes were incubated in pre-defined concentrations of the crude root-peel extract, genistein and praziquantel (as reference drug), till the paralysis time with simultaneous maintenance of respective controls. In the parasite tissue, a significant amount of Ca2+ (approximately 400 microg/g dry tissue wt) was found to be present besides magnesium, iron, zinc, lead and chromium, whilst manganese, cadmium and nickel were below the level of detection. The Ca2+ concentration was decreased significantly by 39%-49%, in the parasite tissue exposed to the test materials in comparison to the respective controls. There was also an increase in Ca2+ efflux by 91%-160% into the culture medium under similar treatments. The changes in Ca2+ homeostasis may be related to the rapid muscular contraction and consequent paralysis in the parasite due to the anthelmintic stress caused by the phytochemicals of F. vestita.


Unexpected serum phosphorus lost in an amniotic fluid embolism patient.
Serum phosphorus concentration reflects body energy equilibrium and functions of the kidney and the coagulation system. It is regulated by serum calcium concentration and parathyroid hormone (PTH). We present a case of constant low concentrations of serum phosphorus in a 34-year-old female who was diagnosed with amniotic fluid embolism (AFE) and was continuously treated with extracorporeal membrane oxygenation (ECMO), continuous renal replacement therapy (CRRT) and blood component transfusion during an observational period of 11 days. This case highlights that the release of inorganic phosphorus from platelets and plasma into the blood prompts PTH secretion. The administration of active vitamin D supplement and PTH antagonism should be considered to neutralize the negative regulatory effect of PTH on serum phosphorus and to benefit patients' recovery in the intensive care unit.


The "right story" to the "right person": communication issues in end-of-life care for Indigenous people.
To explore communication issues faced by health care workers and Indigenous patients and their families in a palliative care setting. Effective communication with Aborigines is especially important because Aboriginal beliefs of health and sickness are so different from Western views. Data were collected from 72 qualitative interviews conducted throughout the regional, rural and remote areas of the Northern Territory with Indigenous patients and carers and the health professionals who care for them. Participants highlighted the struggle associated with effective communication when working in a cross-cultural setting at the interface of Indigenous and Western health care. The findings record the wisdom and insight from practitioners who have extensive experience dealing with communication difficulties.


Afferentation of the lateral nidopallium: A tracing study of a brain area involved in sexual imprinting in the zebra finch (Taeniopygia guttata).
The lateral forebrain of zebra finches that comprises parts of the lateral nidopallium and parts of the lateral mesopallium is supposed to be involved in the storage and processing of visual information acquired by an early learning process called sexual imprinting. This information is later used to select an appropriate sexual partner for courtship behavior. Being involved in such a complicated behavioral task, the lateral nidopallium should be an integrative area receiving input from many other regions of the brain. Our experiments indeed show that the lateral nidopallium receives input from a variety of telencephalic regions including the primary and secondary areas of both visual pathways, the globus pallidus, the caudolateral nidopallium functionally comparable to the prefrontal cortex, the caudomedial nidopallium involved in song perception and storage of song-related memories, and some parts of the arcopallium. There are also a number of thalamic, mesencephalic, and brainstem efferents including the catecholaminergic locus coeruleus and the unspecific activating reticular formation. The spatial distribution of afferents suggests a compartmentalization of the lateral nidopallium into several subdivisions. Based on its connections, the lateral nidopallium should be considered as an area of higher order processing of visual information coming from the tectofugal and the thalamofugal visual pathways. Other sensory modalities and also motivational factors from a variety of brain areas are also integrated here. These findings support the idea of an involvement of the lateral nidopallium in imprinting and the control of courtship behavior.


Heparin-induced thrombocytopenia (HIT II) in liver transplant recipients: a retrospective multivariate analysis of prognostic factors.
We investigated the prevalence of HIT II in liver transplant recipients and analysed associated factors. In recipients with clinically suspected HIT II in the 4Ts pretest clinical scoring system HIPA-assay was performed. Next, 37 clinical variables were analysed retrospectively for their association with HIT II. Factors significantly correlated to our findings in univariate analysis were included in a multivariate model and binary logistic regression analysis. Among 46 recipients 21 patients were suspicious in the 4Ts pretest and 14 of them (30.4%) were diagnosed HIT-antibody positive. Patient's age (P = 0.001), postoperative dialysis (P = 0.028), and postoperative hospital stay (P = 0.035) were significantly associated with development of HIT-antibodies in univariate analysis. Postoperative dialysis and postoperative hospital stay turned out as epiphenomena of patient's age, the only independent predictor (P = 0.021). Using multiple χ(2) -testing, a cut-off could be calculated, assigning patients younger than 59 years to a low risk group and patients of 59 years and older to a high risk group. High incidence of peri-operative HIT II seroconversion in liver transplant recipients is not associated with factors known to induce thrombocyte activation, like blood products or cell-saver. Only patients' age was identified as independent predictor.


Rotational Salter-Harris type 1 fracture of the proximal phalanx.
A purely rotational Salter-Harris type I fracture of the proximal phalanx of the middle finger is described. This injury presents diagnostic problems as the radiographs do not show the displacement. Accurate clinical examination is essential, in particular checking for rotation of the finger.


Imaging treated prostate cancer.
In patients with a clinical suspicion of recurrence after treatment for prostate cancer, imaging can be used to distinguish between local recurrence and metastatic disease. Multiparametric magnetic resonance imaging (mpMRI) of the prostate may be a valuable imaging modality for the detection and localization of local recurrence in patients treated for prostate cancer. In mpMRI, morphological T2-weighted images are combined with functional MRI techniques including diffusion-weighted imaging, dynamic contrast-enhanced imaging, and magnetic resonance spectroscopic imaging to improve accuracy. In this paper, the current status of imaging techniques used to detect and to localize tumor recurrence in patients treated for prostate cancer will be reviewed, with emphasis on mpMRI for local prostate cancer recurrence.


CRISPR/Cas9-mediated tetra-allelic mutation of the 'Green Revolution' SEMIDWARF-1 (SD-1) gene confers lodging resistance in tef (Eragrostis tef).
Tef is a staple food and a valuable cash crop for millions of people in Ethiopia. Lodging is a major limitation to tef production, and for decades, the development of lodging resistant varieties proved difficult with conventional breeding approaches. We used CRISPR/Cas9 to introduce knockout mutations in the tef orthologue of the rice SEMIDWARF-1 (SD-1) gene to confer semidwarfism and ultimately lodging resistance. High frequency recovery of transgenic and SD-1 edited tef lines was achieved in two tef cultivars by Agrobacterium-mediated delivery into young leaf explants of gene editing reagents along with transformation and regeneration enhancing morphogenic genes, BABY BOOM (BBM) and WUSCHEL2 (WUS2). All of the 23 lines analyzed by next-generation sequencing had at least two or more alleles of SD-1 mutated. Of these, 83% had tetra-allelic frameshift mutations in the SD-1 gene in primary tef regenerants, which were inherited in subsequent generations. Phenotypic data generated on T1 and T2 generations revealed that the sd-1 lines have reduced culm and internode lengths with no reduction in either panicle or peduncle lengths. These characteristics are comparable with rice sd-1 plants. Measurements of lodging, in greenhouse-grown plants, showed that sd-1 lines have significantly higher resistance to lodging at the heading stage compared with the controls. This is the first demonstration of the feasibility of high frequency genetic transformation and CRISPR/Cas9-mediated genome editing in this highly valuable but neglected crop. The findings reported here highlight the potential of genome editing for the improvement of lodging resistance and other important traits in tef.


Intra-aneurysmal hemodynamic alterations by a self-expandable intracranial stent and flow diversion stent: high intra-aneurysmal pressure remains regardless of flow velocity reduction.
Little is known about how much protection a flow diversion stent provides to a non-thrombosed aneurysm without the adjunctive use of coils. A three-dimensional anatomically realistic computation aneurysm model was created from the digital subtraction angiogram of a large internal carotid artery-ophthalmic artery aneurysm which could have been treated with either a neck bridging stent or a flow diversion stent. Three-dimensional computational models of the Neuroform EZ neck bridging stent and Pipeline embolization device were created based on measurements with a stereo-microscope. Each stent was placed in the computational aneurysm model and intra-aneurysmal flow structures were compared before and after placement of the stents. Computational fluid dynamics were performed by numerically solving the continuity and Navier-Stokes momentum equations for a steady blood flow based on the finite volume method. Blood was assumed as an incompressible Newtonian fluid. Vessel walls were assumed to be rigid, and no-slip boundary conditions were applied at the lumens. To estimate the change in the intra-aneurysmal pressures we assumed that, at the inlets, the intra-arterial pressure at peak systole was 120 mm Hg both before and after stent placement Without any stent, the blood flow entered into the aneurysm dome from the mid to proximal neck area and ascended along the distal wall of the aneurysm. The flow then changed its direction anteriorly and moved along the proximal wall of the aneurysm dome. In addition to the primary intra-aneurysmal circulation pattern, a counterclockwise vortex was observed in the aneurysm dome. The placement of a Neuroform EZ stent induced a mean reduction in flow velocity of 14% and a small change in the overall intra-aneurysmal flow pattern. The placement of a Pipeline device induced a mean reduction in flow velocity of 74% and a significant change in flow pattern. Despite the flow velocity changes, Neuroform EZ and Pipeline devices induced reductions in intra-aneurysmal pressure of only 4 mm Hg and 8 mm Hg, respectively. The flow diversion effects of both stents were limited to flow velocity reduction. In a non-thrombosed aneurysm or an aneurysm with delayed thrombosis, the intra-aneurysmal pressure remains essentially unchanged regardless of the level of the intra-aneurysmal flow velocity reduction induced by the stents.


More uninsured, fewer excuses. Policy analysts and politicians predict the fallout from recent Census Bureau figures showing a 4.9% increase in uninsured.
With the latest round of Census Bureau data pegging the number of uninsured in the U.S. at 47 million and rising-including 8.7 million children--experts are weighing what the results will mean for the 2008 election. "I think the take-away message is that neither side should be closer to walking away from the debate after looking at these numbers," says James Tallon Jr., right.


Construction of a panel of chromosome-specific oligonucleotide probes (PRINS-primers) useful for the identification of individual human chromosomes in situ.
We present the sequences of a set of oligonucleotides that, when used as primers for PRimed IN Situ (PRINS) labeling, are diagnostic for repetitive sequences in specific human chromosomes. Combined, they enable identification of all human chromosomes except 6, 19, and 20. However, as is also the case with cloned centromeric hybridization probes, chromosomes 14 and 22 are stained together. Along with the sequences of these oligonucleotides we offer a simple, universal procedures for their use. We also present an oligonucleotide that binds to both strands of alpha-satellite DNA, making it possible to specifically amplify alpha-satellite DNA by PCR with this one oligonucleotide alone as primer. The origin of the alpha-satellite DNA in the starting material (whether somatic cell hybrids, flow-sorted chromosomes, or microdissected material) can then be determined on test metaphase spreads by in situ hybridization or PRINS with the PCR product.


Mediation effect of maternal triglyceride and fasting glucose level on the relationship between maternal overweight/ obesity and fetal growth: a prospective cohort study.
Previous studies have suggested that maternal overweight/obesity is asscociated with macrosomia. The present study aimed to investigate the mediation effects of fasting plasma glucose (FPG) and maternal triglyceride (mTG) in the relationship between maternal overweight/obesity and large for gestational age (LGA) among non-diabetes pregnant women. This prospective cohort study was conducted in Shenzhen from 2017 to 2021. A total of 19,104 singleton term non-diabetic pregnancies were enrolled form a birth cohort study. FPG and mTG were measured at 24-28 weeks. We analyzed the association of maternal prepregancy overweight/obesity with LGA and mediation effects of FPG and mTG. Multivariable logistic regression analysis and serial multiple mediation analysis were performed. The odds ratio (OR) and 95% confidence intervals (CIs) were calculated. Mothers who were overweight or obese had higher odds of giving birth to LGA after adjusting potential confounders (OR:1.88, 95%CI: 1.60-2.21; OR:2.72, 95%CI: 1.93-3.84, respectively). The serial multiple mediation analysis found prepregnancy overweight can not only have a direct positive effect on LGA (effect = 0.043, 95% CI: 0.028-0.058), but also have an indirect effect on the LGA through two paths: the independent mediating role of FPG (effect = 0.004, 95% CI: 0.002-0.005); the independent mediating role of mTG (effect = 0.003,95% CI: 0.002-0.005). The chain mediating role of FPG and mTG has no indirect effect. The estimated proportions mediated by FPG and mTG were 7.8% and 5.9%. Besides, the prepregnancy obesity also has a direct effect on LGA (effect = 0.076; 95%CI: 0.037-0.118) and an indirect effect on LGA through three paths: the independent mediating role of FPG (effect = 0.006; 95%CI: 0.004-0.009); the independent mediating role of mTG (effect = 0.006; 95%CI: 0.003-0.008), and the chain mediating role of FPG and mTG (effect = 0.001; 95%CI: 0.000-0.001). The estimated proportions were 6.7%, 6.7%, and 1.1%, respectively. This study found that in nondiabetic women, maternal overweight/obesity was associated with the occurence of LGA, and this positive association was partly mediated by FPG and mTG, suggesting that FPG and mTG in overweight/obese nondiabetic mothers deserve the attention of clinicians.


The biomechanical advantage of lordosis and hip extension for upright activity. Man as compared with other anthropoids.
The mechanics of the muscle-ligament system for a weight lift in man is described. The adaptations of the human spine and the hip to this mechanism are indicated. The efficiency of this mechanism in the human is compared with other primates showing the vast superiority of man in this aspect over his nearest relatives.


Glomerular basement membrane as a compressible ultrafilter.
The ultrafiltration properties of isolated glomerular basement membrane were studied in vitro by forming membrane fragments into thin films for use as ultrafiltration membranes. The filtration properties of the films were examined using cytochrome c, myoglobin, lysozyme, ovalbumin, lactoglobulin, and serum albumin. The films behaved as compressible filters showing size-dependent rejection of the proteins. The behavior of the films was modelled using the fiber matrix hypothesis which gave good prediction of film behavior. The membrane behaved as a random fiber matrix composed of fibers of 0.8-1.0 nm in radius.


Further assessments of ligase LplA-mediated modifications of proteins in vitro and in cellulo.
Posttranslational modifications of proteins are catalyzed by a large family of enzymes catalyzing many chemical modifications. One can hijack the natural use of those enzymes to modify targeted proteins with synthetic chemical moieties. The lipoic acid ligase LplA mutants can be used to introduce onto the lysine sidechain lipoic acid moiety synthetic analogues. Substrate protein candidates of the ligase must obey a few a priori rules. In the present report, we technically detailed the use of a cell line stably expressing both the ligase and a model protein (thioredoxin). Although the goal can be reach, and the protein visualized in situ, many experimental difficulties must be fixed. The sequence of events comprises (i) in cellulo labeling of the target protein with a N3-lipoic acid derivative catalyzed by the mutant ligase, (ii) the further introduction by click chemistry onto this lysine sidechain of a fluorophore and (iii) the following of the labeled protein in living cells. One of the main difficulties was to assess the click chemistry step onto the living cells, because images from both control and experimental cells were similar. Alternatively, we describe at that stage, the preferred use of another technique: the Halo-Tag one that led to the obtention of clear images of the targeted protein in its cellular context. Although the ligase-mediated labeling of protein in situ is a rich domain for which many cellular tools must be developed, many difficulties must be considered before entering a systematic use of this approach. In the present contribution, we added several steps of analytical characterization, both in vitro and in cellulo that were previously lacking. Furthermore, we show that the use of the click chemistry should be manipulated with care, as the claimed specificity might be not complete whenever living cells are used. Finally, we added another approach-the Halo Tag-to complete the previously suggested approaches for labelling proteins in cells, as we found difficult to strictly apply the previously reported methodology.


Modeling the Influence of Vaccine Administration on COVID-19 Testing Strategies.
Vaccination is considered the best strategy for limiting and eliminating the COVID-19 pandemic. The success of this strategy relies on the rate of vaccine deployment and acceptance across the globe. As these efforts are being conducted, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is continuously mutating, which leads to the emergence of variants with increased transmissibility, virulence, and resistance to vaccines. One important question is whether surveillance testing is still needed in order to limit SARS-CoV-2 transmission in a vaccinated population. In this study, we developed a multi-scale mathematical model of SARS-CoV-2 transmission in a vaccinated population and used it to predict the role of testing in an outbreak with variants of increased transmissibility. We found that, for low transmissibility variants, testing was most effective when vaccination levels were low to moderate and its impact was diminished when vaccination levels were high. For high transmissibility variants, widespread vaccination was necessary in order for testing to have a significant impact on preventing outbreaks, with the impact of testing having maximum effects when focused on the non-vaccinated population.


Addition of Co(2+) to culture medium decides the functional expression of a recombinant nitrile hydratase in Escherichia coli.
A nitrile hydratase (NHase) gene from Aurantimonas manganoxydans, cloned and expressed in Escherichia coli, gave an enzyme that efficiently hydrated 3-cyanopyridine to nicotinamide with high thermal stability. We have now found that adding Co(2+) at 0.1 mM to LB medium was essential for production of an active enzyme. However, ≥0.3 mM Co(2+) inhibited the growth of host cells in LB medium and decreased the production of the recombinant NHase. Furthermore, β-mercaptoethanol promoted regeneration of the Co(2+)-defective apoenzyme in vitro possibly by breaking a key disulfide bond thereby promoting the incorporation of Co(2+) into the apoenzyme.


Eyes on the needle: Identification and confirmation of the epidural space.
Epidural catheters are used to provide effective intraoperative and postoperative analgesia. Standard epidural catheterization techniques rely on palpation of surface anatomy and the experience of the anesthesiologist. Failure to correctly place an epidural catheter can lead to inadequate analgesia and serious complications, such as dural puncture headache. Exciting new devices and techniques are being developed for identification of the epidural space and confirmation of catheter entry. This article reviews and describes the recent research findings. The devices and techniques are categorized into three sections: devices that modify the loss of resistance technique; visual confirmation using the epidural needle; and confirmation of placement of the epidural catheter.


Convergent evolution of plant pattern recognition receptors sensing cysteine-rich patterns from three microbial kingdoms.
The Arabidopsis thaliana Receptor-Like Protein RLP30 contributes to immunity against the fungal pathogen Sclerotinia sclerotiorum. Here we identify the RLP30-ligand as a small cysteine-rich protein (SCP) that occurs in many fungi and oomycetes and is also recognized by the Nicotiana benthamiana RLP RE02. However, RLP30 and RE02 share little sequence similarity and respond to different parts of the native/folded protein. Moreover, some Brassicaceae other than Arabidopsis also respond to a linear SCP peptide instead of the folded protein, suggesting that SCP is an eminent immune target that led to the convergent evolution of distinct immune receptors in plants. Surprisingly, RLP30 shows a second ligand specificity for a SCP-nonhomologous protein secreted by bacterial Pseudomonads. RLP30 expression in N. tabacum results in quantitatively lower susceptibility to bacterial, fungal and oomycete pathogens, thus demonstrating that detection of immunogenic patterns by Arabidopsis RLP30 is involved in defense against pathogens from three microbial kingdoms.