The role of bone marrow-derived adult stem cells in a transgenic mouse model of allergic asthma.
Asthmatic airway remodeling is an abnormal injury/repair process of the small airways caused by chronic inflammation in which the quantities of multiple cells increase dramatically. However, the origin of these proliferative cells is still undetermined. The aim of this study was to examine whether bone marrow (BM)-derived adult stem cells are responsible for the proliferative cells in asthmatic airway remodeling. Adult mice were durably engrafted with BM isolated from green fluorescent protein (GFP) transgenic mice. Using GFP BM chimera mice, an ovalbumin (OVA)-induced chronic asthma mouse model was established. The distribution of BM-derived GFP+ cells in the lungs of chronic asthma mice was detected by fluorescence microscopy. The phenotype of BM-derived GFP+ cells in the lung tissues of chronic asthma mice was analyzed by flow cytometry. BM chimera mice were successfully generated, with no detectable radioactive inflammation observed. Using BM chimera mice, we established a mouse model of chronic asthma characterized by a significant increase in the thickness of the airway subepithelial basement membrane and smooth muscle layers. OVA treatment caused many GFP+ cells to appear at sites of small airway inflammation. The extravascular localization of some GFP+ cells and their morphology were not consistent with leukocytes. Flow cytometric analysis of lung cells revealed a significant increase in type I collagen (Col I)+GFP+ cells and α-smooth muscle actin (α-SMA)+GFP+ cells in OVA-treated GFP BM chimera mice. Considerable numbers of Col I- and α-SMA-producing cells originated from BM in the lung tissues of mice with OVA-induced chronic asthma.


Disinfection by-products in Croatian drinking water supplies with special emphasis on the water supply network in the city of Zagreb.
The occurrence of disinfection by-products (DBPs) was investigated in 48 drinking water systems across Croatia. Eleven DBPs were studied: chlorite (ClO2-), chlorate (ClO3-), four trihalomethanes (THMs), and five haloacetic acids (HAAs). Furthermore, an intensive sampling program was conducted in the distribution system in the city of Zagreb where, aside from DBP analyses, natural organic matter (NOM) was characterized using fluorescence spectroscopy. In the drinking waters examined across Croatia, DBP values were found in the range from 0.7 μg/L to 32.8 μg/L for THMs, below LOQ to 17.2 μg/L for HAAs (primarily di- and trichloroacetic acids), below LOQ to 720 μg/L for ClO2- and below LOQ to 431 μg/L for ClO3-. The results obtained showed higher chlorite concentrations in the systems treated with hypochlorite compared to systems treated with chlorine dioxide. DBPs in the Zagreb distribution network were generally low (the average values were below 6 μg/L and 2 μg/L for total THM and total HAA respectively). In contrast to our observations throughout Croatia, dibromoacetic acid (DBAA) was found to be the predominant HAA within Zagreb, most likely due to the degradation of chlorinated carboxylates (di-/tri-chloroacetic) in the network. Characterization of NOM by Parallel Factor Analysis (PARAFAC) fluorescence spectroscopy across the Zagreb network showed distinct temporal variations arising from groundwater inputs, as evident from variable humic-, tyrosine-, and tryptophan-like peaks. Statistical correlations between fluorescence data and DBPs highlight its potential for monitoring the presence of DBPs in distribution networks.


One-lung flooding reduces the ipsilateral diaphragm motion during mechanical ventilation.
Diaphragm motion during spontaneous or mechanical respiration hinders image-guided percutaneous interventions of tumours in lung and upper abdomen. Motion-tracking methods can be applied but increase procedure complexity and procedure time. One-lung flooding (OLF) generates a suitable acoustic pathway to lung tumours and likely suppress diaphragm motion. The aim of this study was to quantify the effect of OLF on ipsilateral diaphragm motion during contralateral one-lung ventilation. To measure the diaphragm motion, M-mode ultrasonography of the right hemidiaphragm was performed during spontaneous breathing and mechanical ventilation, as well as after right-side lung flooding, in three pigs. Diaphragm motion was analysed using magnetic resonance images during left-side lung flooding and mechanical ventilation, in four pigs. Double-lung ventilation increased the diaphragm movement in comparison with spontaneous breathing (17.8 ± 4.4 vs. 12.2 ± 3.4 mm, p = 0.014). Diaphragm movement on the flooded side during contralateral one-lung ventilation was significantly reduced compared to that during double-lung ventilation (3.9 ± 1.0 vs. 17.8 ± 4.4 mm, p = 0.041). By analysing the magnetic resonance images, the hemidiaphragm on the flooded side showed an average displacement of 4.2 mm, a maximum displacement of 15 mm close to the ventilated lung and no displacement at the lateral side. OLF leads to a drastic reduction of diaphragm motion on the ipsilateral side which implies that targeting and motion compensation algorithms for interventions like high-intensity focused ultrasound ablation of intrapulmonary and hepatic lesions might not be required.


HIV-1 activates oxidative phosphorylation in infected CD4 T cells in a human tonsil explant model.
Human immunodeficiency virus type 1 (HIV-1) causes a chronic, incurable infection leading to immune activation and chronic inflammation in people with HIV-1 (PWH), even with virologic suppression on antiretroviral therapy (ART). The role of lymphoid structures as reservoirs for viral latency and immune activation has been implicated in chronic inflammation mechanisms. Still, the specific transcriptomic changes induced by HIV-1 infection in different cell types within lymphoid tissue remain unexplored. In this study, we utilized human tonsil explants from healthy human donors and infected them with HIV-1 ex vivo. We performed single-cell RNA sequencing (scRNA-seq) to analyze the cell types represented in the tissue and to investigate the impact of infection on gene expression profiles and inflammatory signaling pathways. Our analysis revealed that infected CD4+ T cells exhibited upregulation of genes associated with oxidative phosphorylation. Furthermore, macrophages exposed to the virus but uninfected showed increased expression of genes associated with the NLRP3 inflammasome pathway. These findings provide valuable insights into the specific transcriptomic changes induced by HIV-1 infection in different cell types within lymphoid tissue. The activation of oxidative phosphorylation in infected CD4+ T cells and the proinflammatory response in macrophages may contribute to the chronic inflammation observed in PWH despite ART. Understanding these mechanisms is crucial for developing targeted therapeutic strategies to eradicate HIV-1 infection in PWH.


[Effect of a new benzodiazepine derivative on experimental malignant melanomas].
It was found, that (+)cis-3,4-dimethoxy-10,11-dimethyl-6,6aR,7,8,13, 13aS-hexahydro-[I]-benzopyrano-[4,3-b]-1,5-benzodiazepine (ZIMET 54/79) increased the life span (ILS) of C57BL/6/Jena mice suffering from transplanted B16 melanoma (ip.) by 57% (9 times ip. 250 mg/kg) and 90% (9 times ip. 500 mg/kg) respectively. In B6D2F1/Bln mice with transplanted B16 melanoma there was no ILS registered when a dose of 100-500 mg/kg was applied (ip. and p.o.). Using the Harding Passey melanoma (B6D2F1/Bln mice) only 59% tumour volume inhibition (500 mg/kg) without ILS was obtained. Finally ZIMET 54/79 tested on AMel 3 hamster melanoma (strain Z3) decreased the mean tumour volume by 72% (125 mg/kg).


MBO clears way for humanistic approach.
Social worker became patient service counselor, classification clerk became patient service interviewer, and together they became a team with a more humanistic philosophy.


Gut microbial metabolites in Parkinson's disease: Association with lifestyle, disease characteristics, and treatment status.
There is growing appreciation of the importance of the intestinal microbiota in Parkinson's disease (PD), and one potential mechanism by which the intestinal microbiota can communicate with the brain is via bacteria-derived metabolites. In this study, plasma levels of bacterial-derived metabolites including trimethylamine-N-oxide (TMAO), short chain fatty acids (SCFA), the branched chain fatty acid isovalerate, succinate, and lactate were evaluated in PD subjects (treatment naïve and treated) which were compared to (1) population controls, (2) spousal / household controls (similar lifestyle to PD subjects), and (3) subjects with multiple system atrophy (MSA). Analyses revealed an increase in the TMAO pathway in PD subjects which was independent of medication status, disease characteristics, and lifestyle. Lactic acid was decreased in treated PD subjects, succinic acid positively correlated with disease severity, and the ratio of pro-inflammatory TMAO to the putative anti-inflammatory metabolite butyric acid was significantly higher in PD subjects compared to controls indicating a pro-inflammatory shift in the metabolite profile in PD subjects. Finally, acetic and butyric acid were different between PD and MSA subjects indicating that metabolites may differentiate these synucleinopathies. In summary, (1) TMAO is elevated in PD subjects, a phenomenon independent of disease characteristics, treatment status, and lifestyle and (2) metabolites may differentiate PD and MSA subjects. Additional studies to understand the potential of TMAO and other bacterial metabolites to serve as a biomarker or therapeutic targets are warranted.


Imprint cytology versus frozen section: intraoperative analysis of sentinel lymph nodes in breast cancer.
Intraoperative analysis of the sentinel node status in breast cancer adds the benefit of proceeding to axillary dissection at the same time as the primary breast operation, without the need for a second trip to theatre. However, the method of intra-operative analysis must be both rapid and accurate for use in this setting. A prospective series of 20 consecutive patients in one hospital who underwent sentinel node biopsy was performed. Intraoperative evaluation by imprint cytology and frozen section was compared with the final histopathological results. The time taken for both procedures was recorded. A total of 21 sentinel node procedures was performed. The sentinel node was found in 20 procedures (95%). The average time to perform intraoperative pathological analysis was 17 min (range: 5-35 min). Of the eight positive sentinel nodes, five were positive on imprint cytology alone and five were positive on frozen section alone (62.5% accuracy). The statistical false negative rate was 18.8% and the 'false reassurance rate' (patients with an initially negative node that was subsequently positive on formal histology who required a second trip to theatre for axillary dissection) was 37.5%. All patients with a negative sentinel node had a negative axilla. Intraoperative analysis of sentinel lymph nodes in breast cancer can be performed quickly. Imprint cytology and frozen section show comparable accuracy in predicting sentinel node status.


Eclampsia in Italy: A prospective population-based study (2017-2020).
To estimate the incidence, and to investigate risk factors, management, and perinatal outcomes of eclampsia in Italy. This is a prospective population-based cohort study enrolling all women admitted for eclampsia between November 2017 and March 2020. Incident cases were reported using electronic and anonymous data collection forms. Incidence, risk factors, maternal and perinatal morbidity, and mortality. 109 cases were included for an estimated incidence rate of 1.5 (95 % CI 1.2-1.8) per 10.000 births. Risk of developing eclampsia was associated with multiple pregnancies (RR = 4.51; p < 0.001) and with pregnancies achieved with assisted reproductive technologies (RR = 3.03; p < 0.001). Magnesium sulfate was used as prophylaxis in almost 30 % of women with preeclampsia, and to treat an eclamptic fit in 89 % of women. The time interval between the first fit and delivery was 62 min for antepartum and 10 min for intrapartum cases. Around one third of women developed at least one other major complication and one mother died. Severe morbidity affected 13.3 % of the newborns. Two fetal and one neonatal death were reported. Our data revealed low incidence of eclampsia in Italy and prompt administration of antihypertensive drugs and magnesium sulfate to prevent eclampsia and to treat the recurrence of seizures. However, the rate of severe maternal complications is still high: increasing the time interval between fit and delivery seems to be crucial to achieve an effective stabilization of maternal conditions and reduce maternal major complications.


A mutational analysis of the acetylcholine receptor channel transmitter binding site.
Mutagenesis and single-channel kinetic analysis were used to investigate the roles of four acetylcholine receptor channel (AChR) residues that are candidates for interacting directly with the agonist. The EC50 of the ACh dose-response curve was increased following alpha-subunit mutations Y93F and Y198F and epsilon-subunit mutations D175N and E184Q. Single-channel kinetic modeling indicates that the increase was caused mainly by a reduced gating equilibrium constant (Theta) in alphaY198F and epsilonD175N, by an increase in the equilibrium dissociation constant for ACh (KD) and a reduction in Theta in alphaY93F, and only by a reduction in KD in epsilonE184Q. This mutation altered the affinity of only one of the two binding sites and was the only mutation that reduced competition by extracellular K+. Additional mutations of epsilonE184 showed that K+ competition was unaltered in epsilonE184D and was virtually eliminated in epsilonE184K, but that neither of these mutations altered the intrinsic affinity for ACh. Thus there is an apparent electrostatic interaction between the epsilonE184 side chain and K+ ( approximately 1.7kBT), but not ACh+. The results are discussed in terms of multisite and induced-fit models of ligand binding to the AChR.


Studies in fetal wound healing: I. A factor in fetal serum that stimulates deposition of hyaluronic acid.
Fetal wound healing without scar formations, fibrosis, or contracture might be accompanied by major differences in the wound extracellular matrix. The matrix of fetal wounds is rich in hyaluronic acid, a glycosaminoglycan found in high concentrations whenever there is tissue proliferation, regeneration, and repair. Although hyaluronic acid is a critical molecule for both embryonic development and wound healing, no factor has yet been identified that modulates hyaluronic acid in a consistent manner. We describe here a substance present in fetal sheep serum that stimulates hyaluronic acid synthesis by cultured fibroblasts. This glycoprotein factor appears to be ubiquitous, present in fetal sheep and bovine serum, reaching a peak in each at 40% of the way through gestation. This factor is also present in amniotic fluid. It might control hyaluronic acid deposition. In turn, hyaluronic acid, by creating an extracellular environment permissive for cell motility and proliferation, might be critical for fetal development. We suggest that the same sequence of events underlie the unique properties observed in fetal wound healing.


Effect of Danqi Buxin Decoction on Chronic Function Indexes and Life Quality in Patients with Chronic Heart Failure of Yang Deficiency Type.
The purpose was to explore the clinical effect of Danqi Buxin decoction on chronic heart failure (CHF) with yang deficiency and its effect on cardiac function and life quality of patients. 106 CHF patients with yang deficiency treated in Jinan Municipal Hospital of Traditional Chinese Medicine from February 2019 to February 2020 were selected as the research objects and divided into the treatment group and reference group according to the odd and even admission numbers, with 53 cases in each group. The reference group was treated with routine antiheart failure drugs, while the treatment group was additionally treated with Danqi Buxin decoction to compare the clinical effect and cardiac function changes between the two groups. The clinical effective rate in the treatment group was significantly higher than that in the reference group (P < 0.05). The TCM symptom scores at T1, T2, and T3 in the treatment group were significantly higher than those in the reference group (P < 0.05). After treatment, the LVEDV levels in both groups were significantly higher than those before treatment, while the BNP levels were significantly lower than those before treatment (P < 0.001). The LVEDV level in the treatment group after treatment was higher than that in the reference group, while the BNP level in the treatment group was significantly lower than that in the reference group (P < 0.001). The life quality scores in the treatment group after treatment were significantly higher than those in the reference group (P < 0.05). Danqi Buxin decoction on the basis of conventional drugs can significantly improve the cardiac function and life quality of CHF patients with yang deficiency type. Its further research is helpful to establish a good treatment plan for CHF patients.


Combination of Histone Deacetylase Inhibitor Panobinostat (LBH589) with β-Catenin Inhibitor Tegavivint (BC2059) Exerts Significant Anti-Myeloma Activity Both In Vitro and In Vivo.
Over the last three decades changes in the treatment paradigm for newly diagnosed multiple myeloma (MM) have led to a significant increase in overall survival. Despite this, the majority of patients relapse after one or more lines of treatment while acquiring resistance to available therapies. Panobinostat, a pan-histone deacetylase inhibitor, was approved by the FDA in 2015 for patients with relapsed MM but how to incorporate panobinostat most effectively into everyday practice remains unclear. Dysregulation of the Wnt canonical pathway, and its key mediator β-catenin, has been shown to be important for the evolution of MM and the acquisition of drug resistance, making it a potentially attractive therapeutic target. Despite concerns regarding the safety of Wnt pathway inhibitors, we have recently shown that the β-catenin inhibitor Tegavivint is deliverable and effective in in vivo models of MM. In this study we show that the combination of low concentrations of panobinostat and Tegavivint have significant in vitro and in vivo anti-MM effects including in the context of proteasome inhibitor resistance, by targeting both aerobic glycolysis and mitochondrial respiration and the down-regulation of down-stream β-catenin targets including myc, cyclinD1, and cyclinD2. The significant anti-MM effect of this novel combination warrants further evaluation for the treatment of MM patients with relapsed and/or refractory MM.


HIV Vpr protein upregulates microRNA-122 expression and stimulates hepatitis C virus replication.
Human immunodeficiency virus (HIV)/hepatitis C virus (HCV) co-infection is characterized by higher serum HCV RNA loads compared with HCV mono-infection. However, the relationship between HIV and HCV replication remains to be clarified. HIV Vpr has been shown to play an essential role in HIV replication. In this study, we aimed to explore the role of Vpr in HCV replication and pathogenesis. We therefore used the genotype 2a full-length HCV strain JFH1 infection system and the genotype 1b full-length HCV replicon OR6 cell line to analyse the effects of Vpr on HCV replication. We found that Vpr promoted HCV 5' UTR activity, HCV RNA replication and HCV protein expression in two HCV infection cell models. Additionally, lymphocyte-produced Vpr significantly induced HCV 5' UTR activity and HCV replication in hepatocytes. We also found that Vpr upregulated the expression of miR-122 by stimulating its promoter activity. Furthermore, an miR-122 inhibitor suppressed the Vpr-mediated enhancement of both HCV 5' UTR activity and HCV replication. In summary, our results revealed that the Vpr-upregulated expression of miR-122 is closely related to the stimulation of HCV 5' UTR activity and HCV replication by Vpr, providing new evidence for how HIV interacts with HCV during HIV/HCV co-infection.


Virtual non-contrast detector-based spectral CT predictably overestimates tissue density for the characterisation of adrenal lesions compared to true non-contrast CT.
To establish if virtual non-contrast (VNC) images generated from contrast-enhanced detector-based spectral CT could replace true non-contrast (TNC) imaging for the characterisation of adrenal masses. TNC and VNC images were retrospectively reviewed for 39 patients with one or more adrenal lesions who underwent contrast-enhanced spectral CT of the upper abdomen. Lesions were categorised as either 'adenoma' or 'indeterminate/other lesion' based on current reference standards. The CT density of each lesion was measured on both image sets by two readers and compared using Wilcoxon signed-rank test. ROC analysis with Youden's J index method was performed to determine the optimal attenuation cut-off for diagnosing benign adenoma on VNC images. Forty-four lesions were included, 37 of which were diagnosed as adenomas. There were significant differences between TNC and VNC measurements for both readers (mean difference 9.1 HU for reader 1; 9.8 HU for reader 2; p < 0.01). Optimal attenuation thresholds for diagnosing adenomas on VNC were 25.3 HU (reader 1) and 23.9 HU (reader 2) for the entire population, and 18.3 HU (reader 1) and 19.7 HU (reader 2) for lipid-rich adenomas < 10 HU on TNC imaging. There is insufficient evidence to support the use of VNC as a substitute for TNC images in the characterisation of adrenal lesions. VNC using a detector-based spectral CT scanner shows a predictable increase in attenuation values compared to TNC. Thus, future studies might be better directed towards finding a new threshold value for diagnosing benign adrenal adenomas on VNC imaging.


Alternatives to conventional ileostomy in chronic ulcerative colitis.
As long as surgeons have performed total proctocolectomy with permanent ileostomy, efforts have been made to avoid the necessity for a permanent stoma. Preservation of sphincter function can now be achieved by more liberal application of the operation of abdominal colectomy and ileorectal anastomosis in carefully selected patients, and knowledge of the progression from dysplasia to cancer can be applied in a prospective fashion to those persons potentially at risk who have retained rectal mucosa. Individuals who are incapable of dealing with conventional permanent ileostomy and who lack the potential use of their own sphincters can be offered the alternative of a continent ileostomy. The advantages of both of these procedures can be combined by greater application of the operation of total abdominal colectomy, mucosal proctectomy, and ileoanal reservoir with ileoanal anastomosis. Although enthusiasm exists for all of these procedures, their results should be compared with the results of what must still be considered the "gold standard" operation, that is, proctocolectomy and conventional ileostomy. Nevertheless, the alternative procedures represent a considerable advance in the overall care of patients with chronic ulcerative colitis.


Ectopic atrial rhythm after superior septal approach to the mitral valve - an often under-reported entity.
Very few reports elaborate on the changes in P wave following superior septal approach to the mitral valve. We aimed to describe the changes in the P wave axis and other electrocardiographic changes following this approach among patients preoperatively in sinus rhythm. We did a retrospective review of medical records among all our patients undergoing superior septal approach for mitral valve surgery from September 2014 to September 2019. Electrocardiograms during hospital stay and until 6-month follow-up were analyzed. A deviation in P wave axis from the normal range of + 30 to + 60° was classified as ectopic atrial rhythm. In the study population of 47 patients (age 16-75 years, 51.3 ± 13.6 years; M:F ratio 3.7:1), who were in normal sinus rhythm preoperatively, 34 patients (72.3%) had a visible P wave on electrocardiogram (ECG) at discharge. Among them, the P wave axes of 17 patients (36.2%) were within normal range (normal sinus rhythm), whereas 17 patients (36.2%) had ectopic atrial rhythm at discharge. The most frequent abnormal P wave axis was between 0 and - 30° (12 patients). At 6 months, 8 patients (17.0%) had a persistent ectopic atrial rhythm. These patients underwent a Holter test at 6 months and were followed up for symptomatic bradycardia for 3 years. None of the patients with ectopic atrial rhythm required pacemaker insertion. Persistence of ectopic atrial rhythm at 6 months is common (17%) after superior septal approach. Documentation of P wave axis after this approach will help avoid missing it. These patients may be kept on follow-up to look for symptomatic bradycardia.


Identification and characterization of two isoforms of an endothelin-converting enzyme-1.
We report the cloning and sequencing of 5'-terminal region of a beta form of rat ECE-1 cDNA which is different only in its N-terminal amino-acid sequence to the cDNA we have cloned previously (alpha form [K. Shimada et al. (1994) J. Biol. Chem. 269, 18275-18278]). No significant difference was found in the specific activity and substrate specificity between the two isoforms. The expression level of ECE-1 alpha mRNA was higher than that of ECE-1 beta in various rat cells and tissues, suggesting that the physiologically important isoform is ECE-1 alpha. The present findings verified the presence of two forms of ECE-1 over many species, which are created probably through alternative splicing.


Relation of coronary artery disease in women < 60 years of age to the combined elevation of serum lipoprotein (a) and total cholesterol to high-density cholesterol ratio.
After age 40 years, coronary artery disease (CAD) is the leading cause of death in both women and men, yet in women the factors associated with, or leading to, CAD have been less extensively studied. This study examined the strength of association of a number of risk factors to CAD in groups of women < 60 years of age with (n = 108) and without (n = 66) angiographically documented significant narrowing of coronary arteries. In univariate analyses, there were significant differences between control subjects and patients with regard to age (49 +/- 6 vs 52 +/- 7 years) and total lipids and apolipoproteins measured. The relative frequency of cigarette smoking and diabetes was higher and that of estrogen replacement therapy lower in patients with CAD than in control subjects. In multivariate analysis the following factors were independently associated with CAD (adjusted odds ratios and 95% confidence intervals): total cholesterol to high-density lipoprotein (HDL) cholesterol (1.91; 1.56 to 2.34); lipoprotein (a) (10.66; 3.51 to 32.35); estrogen replacement (0.24; 0.11 to 0.54); age (1.12; 1.04 to 1.18); and smoking (1.50; 0.98 to 2.29). The nonadjusted odds ratio of CAD, based on combined tercile values of lipoprotein (a) serum level and total cholesterol to HDL cholesterol ratio, was very low (0.15; 0.06 to 0.36) when both values were within the first tercile, but very high (16.63; 3.54 to 78.07) when both were in the third tercile.(ABSTRACT TRUNCATED AT 250 WORDS)


Synthesis, anti-tuberculosis activity, and 3D-QSAR study of ring-substituted-2/4-quinolinecarbaldehyde derivatives.
We have previously identified ring-substituted quinolines as a new structural class of anti-tuberculosis agents. In our ongoing efforts at structural optimization of this class, four series of ring-substituted-2/4-quinolinecarbaldehyde derivatives were synthesized. All twenty-four compounds were synthesized using short and convenient one to two high yielding steps. The newly synthesized compounds were tested in vitro against drug-sensitive Mycobacterium tuberculosis H37Hv strain. Several derivatives were found to be promising inhibitors of M. tuberculosis. For example, derivatives 4a-c (Series 2), 7a-d (Series 3), and 8a-b (Series 4) displayed >90% inhibition at 6.25 microg/mL in the primary assay. The most active compounds, N-(2-fluorophenyl)-N'-quinolin-2-ylmethylene-hydrazine (4a), N-(2-adamantan-1-yl-quinolin-4-ylmethylene)-N'-(4-fluorophenyl)hydrazine (7c), and N-(2-cyclohexyl-quinolin-4-ylmethylene)-N'-(2-fluorophenyl)hydrazine (8a), exhibited 99% inhibition at the lowest tested concentration of 3.125 microg/mL against drug-sensitive M. tuberculosis H37Rv strain. The similarity index based on steric and electrostatic features of the molecules was used, in conjunction with principal component analysis and linear discriminant analysis, successively to classify the molecules based on their activity into two classes. This classification method gives us confidence in predicting the activity class of any new unsynthesized molecule belonging to these series.


Detection and quantification of foot and mouth disease virus by enzyme labelled immunosorbent assay techniques.
Enzyme labelled immunosorbent assays (ELISA) have been developed to detect and quantify foot and mouth disease (FMD) virus using flexible plastic microtitre plates. The methods were successful for the specific detection of FMD virus and were 50 to 100 times more sensitive than the complement fixation test. The application of the ELISA techniques to FMD virus typing and subtyping, and to the assay of antigen concentration during manufacture of vaccines is discussed.


Alterations in colonic transit time after laparoscopic versus open cholecystectomy: a clinical study.
Postoperative enteral paresis constitutes a common problem for surgeons around the world. Evidence by many authors suggests that colonic inertia constitutes a major component of postoperative enteral paresis. This study aims at comparing the effect of laparoscopic versus open cholecystectomy on colonic transit time in humans. In this study, were included a total of 29 patients suffering from cholelithiasis, divided into two groups, a laparoscopic cholecystectomy and an open cholecystectomy group. All patients ingested one capsule containing 24 radiopaque markers on the day of the operation, and plain abdominal films were obtained on the 3rd postoperative day. The number of remaining markers was counted, and the percentage of rejected markers was calculated. For the statistical analysis, SPSS for windows version 12 was used. The study's results show a significant difference in postoperative colonic motility, in favor of the laparoscopic cholecystectomy group (P = 0,001). Causative interpretation of these results is difficult, mainly due to the multifactorial nature of postoperative colonic hypomotility. The present study suggests an advantage of laparoscopic cholecystectomy, as far as the duration of postoperative colonic paresis is concerned.


Degradation of the arginyl-tRNA synthetase protein during purification by affinity chromatography on immobilized total tRNA and immobilized tRNA, specific for arginyl-tRNA synthetase.
Arginyl-tRNA synthetase was prepared from Bom:NMRI mouse liver postribosomal supernatants. Purification was performed by either gel filtration on Sephadex G-200 or chromatofocusing before affinity chromatography on immobilized total tRNA (mixture of tRNA's, specific for all aminoacyl-tRNA synthetases), followed by affinity chromatography on immobilized tRNA, specific for arginyl-tRNA synthetase. The purification factor for arginyl-tRNA synthetase activity was 16700x. Following sodium dodecyl sulphate polyacrylamide electrophoresis several protein bands were found in the purified material. All these protein bands were shown to possess arginyl-tRNA synthetase activity.


Colonic interposition for benign esophageal disease. Long-term clinical and endoscopic results.
Replacement of the esophagus for benign disease requires familiarity with the long-term results of various esophageal substitutes. In the present study, 60 esophageal reconstructions for benign disease using colonic interposition have been presented. The operations were performed mainly without thoracotomy, using both antiperistaltic and isoperistaltic colonic segments. There were no differences in swallowing ability between patients with antiperistaltic and patients with isoperistaltic interpositions. Regurgitation symptoms, however, seemed to be somewhat more common and more difficult in patients with antiperistaltic colonic transpositions. Endoscopic signs of colitis were common, but they did not correlate with regurgitation symptoms. Bacterial cultures from the transplanted colon mainly revealed the usual mouth organisms. Candida albicans was frequently found in the fungal samples. There were no differences in the results between patients with follow-up periods of more and less than 2 years. The clinical results were good or fair in a great majority of the patients.


Inosine, an endogenous purine nucleoside, avoids early stages of atherosclerosis development associated to eNOS activation and p38 MAPK/NF-kB inhibition in rats.
Atherosclerosis is a multifactorial chronic disease, initiated by an endothelial dysfunction. Adenosine and its analogs can change a variety of inflammatory diseases and has shown important effects at different disease models. Inosine is a stable analogous of adenosine, but its effects in inflammatory diseases, like atherosclerosis, have not yet been studied. The aim of this study was to evaluate the pharmacological properties of inosine, administered sub chronically in a hypercholesterolemic model. Male Wistar rats were divided into four groups: control group (C) and control + inosine (C + INO) received standard chow, hypercholesterolemic diet group (HCD) and HCD + inosine (HCD + INO) were fed a hypercholesterolemic diet. At 31st experimentation day, the treatment with inosine was performed for C + INO and HCD + INO groups once daily in the last 15 days. We observed that the hypercholesterolemic diet promoted an increase in lipid levels and inflammatory cytokines production, while inosine treatment strongly decreased these effects. Additionally, HCD group presented a decrease in maximum relaxation acetylcholine induced and an increase in contractile response phenylephrine induced when compared to the control group, as well as it has presented an enhancement in collagen and ADP-induced platelet aggregation. On the other hand, inosine treatment promoted a decrease in contractile response to phenylephrine, evoked an improvement in endothelium-dependent vasorelaxant response and presented antiplatelet properties. Moreover, inosine activated eNOS and reduced p38 MAPK/NF-κB pathway in aortic tissues. Taken together, the present results indicate inosine as a potential drug for the treatment of cardiovascular disorders such as atherosclerosis.


Glycoprotein gpTRL10 of human cytomegalovirus is dispensable for virus replication in human fibroblasts.
Human cytomegalovirus (HCMV) has a coding capacity for glycoproteins which by far exceeds that of other herpesviruses. Few of these proteins have been characterized. Glycoprotein gpTRL10 represents a structural component of HCMV. The protein has no homologous counterparts in other herpesviruses. We have isolated bacterial artificial chromosomes (BACs) harboring the genome from the low passage clinical isolate PAN and constructed a deletion mutant in reading frame TRL10. Our results show that deletion of gpTRL10 has only minimal effects on viral replication in human fibroblasts.


Tolerance of fat emulsions in very-low-birthweight neonates. Monitoring of plasma lipid concentrations.
Tolerance to parenterally administered fat emulsions was studied in 45 normally grown neonates ranging from 820 to 1550 gm in birthweight, from 27 to 34 weeks in gestational age, and from 2 to 10 days postnatal age. Concentrations of total lipids, free glycerol, true triglycerides, free fatty acids, and cholesterol in plasma were studied over an 8-day period. The aim was to determine whether the concentrations of any one lipid fraction could be used to predict those of other fractions. With 650 pairs of data being correlated, the best correlation coefficients were obtained between total lipids and triglycerides (r = 0.67), total lipids and cholesterol (r = 0.63), free glycerol and free fatty acids (r = 0.55), total lipids and free fatty acids (r = 0.54), and triglycerides and free fatty acids (r = 0.50). Although all correlation coefficients were highly significant statistically (P less than 0.001), the very large standard errors precluded using any of these relationships to make clinical predictions. Problems and limitations of currently available techniques are discussed in order to provide a lead for further research.


Sonographic appearances of medial retinacular complex injury in transient patellar dislocation.
To describe the sonographic appearances of the medial retinacular (MPFR) complex of the knee in patients with acute and recurrent patellar dislocation. Thirty patients were scanned within 2-4 weeks of an acute episode of lateral patellar dislocation. Eleven gave a history of recurrent patellar dislocation. Ten patients had examination under anaesthesia with arthroscopy and repair of the injury. The sonographic and operative results were compared. The normal sonographic appearance of the MPFR is described. Of the 10 patients who underwent examination under anaesthesia, four patients had complete avulsion of the MPFR from the patella, two patients had avulsion of the MPFR from the adductor tubercle and four patients had avulsion of the MPFR from both the patella and adductor tubercle. There was complete correlation between the sonographic and operative findings for injuries of the MPFR. Other findings included partial retinacular tears, injury to the medial collateral ligament, haematoma within vastus medialis obliquus (VMO) and bony avulsions from the patella and adductor tubercle Sonography gives reliable information regarding the site of the injury and its extent thus helping to decide whether conservative or operative treatment is the most appropriate approach to management of the injury.


Tumor growth-promoting cellular host response during liver atrophy after portal occlusion.
Clinical observations suggest cancer progression after preoperative segmental portal vein occlusion, a procedure to prevent liver failure after major hepatic resections. The aim of this study was to determine whether portal occlusion induces host reactions which promote cancer invasion and angiogenesis. The rat model of portal branch ligation (PBL) was compared with partial hepatectomy (PH) and sham operation (SO) and evaluated for the expression of heat shock protein-70 (hsp70), heme oxygenase-1 (hmox1), early growth response gene-1 (Egr-1) and urokinase-type plasminogen activator (uPA), its inhibitor (PAI-1) and receptor (uPAR). Portal deprivation after PBL was associated with a regression of liver tissue to 25% of its original mass within 8 days with only modest fibrotic changes. During the progression of atrophy, there were significant inductions of hsp70-, hmox1- and Egr-1-mRNA in comparison with regenerating liver tissue. PAI-1-specific mRNA was transiently elevated at 3 - 48 h after PBL in the atrophying lobes, whereas uPA and uPAR were unaffected in comparison with PH or SO. Hepatic atrophy caused by PBL is associated with increased expression of genes known to promote tumor growth. These host events represent a possible explanation for the tumor progression after portal occlusion and require further evaluation.


Intraoperative radiation therapy for advanced neuroblastoma: the problem of securing the IORT field.
The purpose of this study is to evaluate the efficacy of intraoperative radiation therapy (IORT) and the problem of securing the IORT field in advanced pediatric neuroblastoma. Between 1996 and 2005, 12 children received IORT for advanced pediatric neuroblastoma patients. Electron beam energies ranged from 10 to 12 MeV and median dose was 10 Gy (8-12 Gy). All of them had surgery with IORT against the primary tumor site and the abdominal aorta surroundings. A gross total resection (GTR) was achieved in 10 patients and subtotal resection (STR) was two patients. All of 12 patients were classified as high risk. Nine patients were alive 17-120 (mean 48 months) after diagnosis. Local tumor control was achieved in 100% of patients, of whom one experienced local recurrence outside the IORT field. At the operation, it was difficult to secure the IORT field because of the angle of the radiation cylinder in three patients. One of the three of these patients experienced local recurrence outside of the IORT field in the upper side of superior mesenteric artery and two of three patients had an external beam radiation after surgery, and there was no local recurrence. One patient had a postoperative ileus, and one patient had transient diarrhea and hydronephrosis. For advanced neuroblastoma patients, IORT produced excellent local control after surgery. However, there is a problem of securing the IORT field. For local control, it is necessary to add an external beam radiation after IORT when it is difficult to secure the IORT field.


Chronic infection in woodchucks infected by a cloned hepatitis delta virus.
Two woodchucks (Marmota monax) intrahepatically inoculated with hepatitis delta virus (HDV) complementary DNA clones pSVL-D3 and pSVL-Ag showed virological and pathological signs of acute and chronic HDV infection. HDV-RNA and hepatitis delta antigen (HDAg) were detected in serum by slot-blot hybridization and by western blot five weeks after inoculation. Liver biopsy specimens collected at 8th week post inoculum were positive for HDV-RNA. Anti-HDV antibodies were detected at the 11th and 9th weeks, respectively. Histological finding of hepatocarcinoma and persistence of circulating HDV-RNA and anti-HDV were observed up to the 10th month. Both woodchucks produced "small" and "large" HDAg antigen, although the inoculated cloned DNA bears the coding capability solely for the small antigen. A transient decrease of woodchuck hepatitis virus DNA (WHV-DNA) level was observed during the peak of HDV infection. Successive inoculation of acute-phase serum in three woodchucks resulted in a successful infection in one of the animals.


Anti-hepatitis E virus antibody prevalence in French expatriate workers.
The hepatitis E virus (HEV) is a leading cause of acute hepatitis in developing countries and an emerging pathogen in Europe. HEV seroprevalence has rarely been assessed in cohorts of travelers, and previous studies have reported a very low rate of exposure. We assessed HEV seroprevalence in French expatriate workers. The prevalence of HEV IgG and IgM was assessed among 43 French expatriate workers using two commercial microplate enzyme immunoassays (Adaltis and Wantai). Additionally HEV IgG-positive sera were tested with an immunoblot assay (recomLine), while IgM-positive sera were tested with a rapid immunochromatographic assay (Assure). The prevalence of anti-HEV IgG was 3.7 times higher in French expatriates than in comparable blood donors from the same area. A discrepancy was evidenced between the HEV IgG results obtained by the Wantai and Adaltis assays (48.8% vs. 30.2%). Expatriation from France, including to areas not recognized as hyperendemic for HEV, may expose individuals to HEV infection. This issue warrants further study; in particular, serology should be compared before and after travel. The most sensitive Wantai serological assay should be used for epidemiological studies to obtain better insight into the epidemiology of HEV.


Muscarinic receptor blockade attenuates reserpine-mediated Fos induction in the rat striatopallidal pathway.
Acute administration of the dopamine-depleting agent reserpine (10 mg/kg) induces Fos expression in striatopallidal neurons of intact rats-an effect that is blocked by pretreatment with the D2 agonist quinpirole (0.5 mg/kg). Systemic administration of the muscarinic antagonist scopolamine (50 mg/kg) partially attenuates reserpine-mediated striatal Fos expression. These data suggest that muscarinic receptors, either within the striatum or in extrastriatal sites, regulate D2 receptor-mediated Fos expression in rat striatopallidal neurons.


Screening for congenital cataracts: a cost-consequence analysis of eye examination at maternity wards in comparison to well-baby clinics.
To estimate, on a national basis in Sweden, the costs versus consequences of combined maternity ward and well-baby clinic eye screening compared to well-baby clinic screening alone. Two scenarios were created and compared regarding healthcare costs: visual acuity development and quality-adjusted life-years (QALYs). One scenario represented early management (combined maternity ward and well-baby clinic screening); the other represented less early management (well-baby clinic screening only). Each scenario was based on 100,000 births, and the healthcare costs were calculated from detection until age 18 y. All estimates of prevalence, visual outcome, postoperative complications and screening procedures were based on previous, recently published studies. Estimated costs were obtained from St. Erik's Eye Hospital in Stockholm and the University Hospital of Lund, at 2001 prices. Total cost of the maternity ward/well-baby clinic screening scenario was 7.9 million SEK, and that of the maternity ward screening scenario was 6.9 million SEK. The incremental cost-effectiveness ratio was estimated at 234,000 SEK/QALY provided three more children per year were detected in Sweden by mandatory maternity ward/well-baby clinic screening. The incremental expense of introducing combined maternity ward/well-baby clinic eye screening on a nationwide basis is cost effective and within acceptable levels of cost/QALY when compared with other widely accepted therapies across diverse medical specialties.


Chorionic villus sampling for first-trimester prenatal diagnosis: Northwestern University program.
We present our initial experience in developing a chorionic villus sampling program at Northwestern University. In phase 1, we performed chorionic villus sampling in 58 patients prior to elective first-trimester abortion, assessing the reliability and reproducibility of obtaining adequate villus samples and performing cytogenetic analysis by means of both the direct and culture methods. Specimens were categorized according to quality: class I, multiple identifiable villi (n = 20); class II, few villi or villi mixed with decidua (n = 15); class III, no villi (n = 23). There was a positive trend between operator experience, amount of villi obtained, and quality of cytogenetic preparations. In March, 1984, we received Institutional Review Board approval to perform chorionic villus sampling in continuing pregnancies (phase 2). Among the first 20 cases we found two abnormalities (47,XY, + 13; 45,X). The remaining 18 pregnancies were continuing. Recommendations are made for developing a chorionic villus sampling program.


Hypoprolactinemia and ovarian function.
Thirty-two patients with ovarian hyperstimulation were randomized to receive bromocriptine or placebo from cycle day 5 onward. Bromocriptine decreased serum and follicular fluid prolactin (PRL), accelerated ovarian follicle growth, increased serum and follicular fluid estradiol, lowered luteal phase progesterone, and shortened the luteal phase length of the cycle. The maximal luteal phase estradiol and progesterone concentrations correlated with each other in the placebo group, but not in the bromocriptine group. These findings indicate that hypoprolactinemia interferes with ovarian function. The unchanged concentrations of gonadotropic hormones and pattern of luteinizing hormone pulsation during bromocriptine suggest direct ovarian effects of hypoprolactinemia. Because PRL suppression enhanced follicular responses and inhibited corpus luteum formation and function, the follicular and corpus luteum actions of PRL may be different.


Rapid granulation of aerobic granular sludge and maintaining its stability by combining the effects of multi-ionic matrix and bio-carrier in a continuous-flow membrane bioreactor.
The present investigation aimed at providing a novel approach to promote the rapid granulation and stability of aerobic granular sludge (AGS) in a continuous-flow membrane bioreactor (MBR). By operating two identical MBRs with or with no bio-carrier for 125 days, it was found that the combination of multi-ionic matrix and bio-carrier could promote the rapid formation and maintain the long-term stability of AGS. The primary AGS was first observed inside the reactor on day 14, and the mature AGS appeared soon and kept stable for more than 4 months (its average size still was about 800 μm on day 125). Suitable filling ratio of bio-carrier was beneficial to form a stable and regular circulating water flow inside, and adding divalent metal ions quickly reduced the negative charges of tiny sludge particles, which were two essential factors leading to the rapid granulation of AGS and maintaining its stability. The multi-ionic matrix not only enhanced the biological aggregation process, but also facilitated the expansion of the cultivated AGS into a new multi-habitat system of Mn-AGS, in which, complex microbial communities with rich bio-diversity robustly promoted the efficient removal of organic pollutants and nutrients.


Problems with surveillance methods for alcoholism: differences in coding systems among federal, state, and private agencies.
Social indicator systems can serve as a social accounting method to guide public policy on alcoholism, utilizing data which are routinely collected at public expense. An attempt to develop an alcoholism social indicator system for Minnesota demonstrated many differences in the coding schemes used by various state agencies and institutions. These findings have relevance to other social indicator systems being developed to assess public policies regarding the people's health.


A Transosseous Suture as an Alternative to Suture Anchor on Anterior-Avulsion Greater Tuberosity Fragment Fixation in Neer Three-Part Proximal Humeral Fracture: A Biomechanical Study.
Greater tuberosity (GT) fragments were communicated, and additional techniques to increase the GT fragment stability after the locking plate fixation was necessary. This study aimed to analyze the reinforcement effects on the anterior-avulsion GT fragment in Neer three-part proximal humeral fractures (PHFs) using transosseous suture and suture anchor techniques. Eighteen fresh-frozen human cadaveric shoulder specimens were used in the study. Standardized fracture of the GT and surgical neck was created in 18 human cadaveric proximal humerus. The GT fragments were reinforced with transosseous suture (TS), suture anchor (SA), and suture in addition to the PHILOS plate fixation. The fixed humerus was tested by applying static loading to the supraspinatus tendon. Load forces and fragment displacement were evaluated by a biomechanical testing machine, and the load to 3- and 5-mm displacements, load to failure, and mode of failure were recorded for all specimens. Nonparametric variables were examined by the Kruskal-Wallis test, and the Bonferroni post hoc test was used to analyze the mean loads to create 3- and 5-mm displacements as well as the failure load. The age, female proportion, and bone mineral density showed no statistically significant differences between the three groups. The mean loading force to create 3-mm and 5-mm displacement in the TS group (254.9 ± 77.4, 309.6 ± 152.7) were significantly higher than those in the suture group (136.1 ± 16.7, 193.4 ± 14.5) (P = 0.024, P = 0.005). For the SA group, the force to create 3- and 5-mm displacement (204.3 ± 60.9, 307.8 ± 73.5) were comparable to those in the TS group (P = 0.236, P = 0.983). Moreover, the loading force to failure in the TS group (508.6 ± 217.7) and SA group (406.6 ± 114.9) was significantly higher than that in the suture group (265.9 ± 52.1) (P = 0.021, P = 0.024). In the TS group, three failed due to tendon-bone junction rupture; bone tunnel broken occurred in two specimens; suture rupture could also be seen in one specimen. All specimens in the suture group failed because of suture rupture. In the SA group, three specimens failed due to suture rupture; two failed secondary to tendon-bone junction rupture; and one failed because of shaft fracture. Transosseous suture is a new type of reinforcement for GT fragment in Neer-three part PHFs. The transosseous suture was superior to the suture only in the reinforcement of the anterior-avulsion GT fragment of Neer three-part PHFs, and it had comparable biomechanical strength to the suture anchor.


Intense, stable and excitation wavelength-independent photoluminescence emission in the blue-violet region from phosphorene quantum dots.
Nanoscale phosphorene quantum dots (PQDs) with few-layer structures were fabricated by pulsed laser ablation of a bulk black phosphorus target in diethyl ether. An intense and stable photoluminescence (PL) emission of the PQDs in the blue-violet wavelength region is clearly observed for the first time, which is attributed to electronic transitions from the lowest unoccupied molecular orbital (LUMO) to the highest occupied molecular orbital (HOMO) and occupied molecular orbitals below the HOMO (H-1, H-2), respectively. Surprisingly, the PL emission peak positions of the PQDs are not red-shifted with progressively longer excitation wavelengths, which is in contrast to the cases of graphene and molybdenum disulphide quantum dots. This excitation wavelength-independence is derived from the saturated passivation on the periphery and surfaces of the PQDs by large numbers of electron-donating functional groups which cause the electron density on the PQDs to be dramatically increased and the band gap to be insensitive to the quantum size effect in the PQDs. This work suggests that PQDs with intense, stable and excitation wavelength-independent PL emission in the blue-violet region have a potential application as semiconductor-based blue-violet light irradiation sources.


Exploration of reasons for feeding choices in Hispanic mothers.
To explore factors that contribute to feeding choices Hispanic women make for their newborn infants in the immediate postpartum period, with a focus on the phenomenon of "los dos," breastfeeding with formula supplementation. A descriptive qualitative design that utilized a semistructured interview guide with open-ended questions was used. Qualitative data preparation and analysis methods were used to identify themes surrounding feeding choices. The sample included 19 women within 48 hours of giving birth who self-identified as Spanish speakers. Reasons included 1) previous experience; 2) "no llora con hambre" (no crying for hunger); 3) "para salud" (for health); and 4) to prevent suffering that might occur when the mother and infant were separated while the mother worked "no quiero que sufra mientras trabajo." Discussion of infant feeding choices should occur prior to birth since many mothers decide on feeding method while still pregnant. At the first prenatal visit identification of preference should occur to include not just breast or formula, but also asking about "los dos." Educational interventions that directly address the "los dos" feeding practice should be culturally appropriate, evaluated and disseminated for use in the prenatal period. Influential family members should be included in prenatal visits and educational sessions to help assure that the woman is not receiving mixed messages at home. Assessment of the workplace environment should also occur during the prenatal period with an exploration of how pumping and storing of breast milk might be possible once women return to work after the baby's birth.


A program for the prevention of lung diseases.
The pathogenesis, morbidity, mortality, and economic impact of chronic obstructive pulmonary disease (COPD) are similar to those of coronary heart disease. But in contrast to cardiologists pulmonologists have not yet developed and tested worldwide preventive strategies. Based on a research pneumobile in the FRG, the Institute for Preventive Medicine in Graz has developed a Pneumobile for general use. The program is aimed at all groups that can contribute to the prevention of COPD. This means the lay public, physicians, health-care providers, and politicians, and especially the media. To date about 10,500 people from all social levels have been tested in the pneumobile; 8% underwent a lysis test. The Pneumobile was stationed at heavily frequented locations in urban and rural centers. About 10 times as many people got a close look, and 100-1000 times as many heard of the Pneumobile and were made aware of the problem of COPD.


Long term oncological outcomes for laparoscopic versus open surgery for rectal cancer - A population-based nationwide noninferiority study.
The aim of this work was to compare the 5-year overall survival in a national cohort of patients undergoing curative abdominal resection for rectal cancer by laparoscopic (LAP) or open (OPEN) surgery. All patients diagnosed with clinical Stage I-III rectal cancer and who underwent LAP or OPEN abdominal curative surgery in Sweden between 2010 and 2016 were retrieved from the Swedish Colorectal Cancer Registry. A noninferiority study design was employed with a statistical power of 90%, a one-side type I error of 2.5% and a noninferiority margin of 2%. The analyses were performed as intention-to-treat and the relationship between surgical technique and overall mortality within 5 years was analysed. Multilevel regression models with the patients matched by propensity scores adjusted for patient- and tumour-related variables were used. A total of 8410 Stage I-III cancer patients were included. This group underwent 2094 LAP (24.9%) and 6316 OPEN (75.1%) procedures and were followed until 31 December 2020. Multivariable Cox regression demonstrated that 5-year overall survival was higher in the LAP group [hazard ratio (HR) 0.877; 95% CI 0.775-0.993]. [Correction added on 21 November 2022, after first online publication: In the preceding sentence, the CI value for LAP group has been corrected from "0.877" to "0.775" in this version.] The outcome was similar when multiple imputation and propensity score matching were employed. When cT4 patients were excluded there was no difference (HR 0.885; 95% CI 0.790-1.033). At 5-years' follow-up local recurrence was not different, at 2.9% for the LAP group and 3.6% for the OPEN group (p = 0.075), while metastatic disease was more frequent in the OPEN group (19.6% compared with 15.6% for LAP; p < 0.001). This study demonstrated that the LAP technique was not inferior to OPEN surgery with regard to overall 5-year survival. These results support the use of laparoscopic surgery.


The associations of urinary DEHP metabolite levels, serum thyroid hormones, and thyroid-related genes among the adolescent students from China: a cross-sectional study.
Our study aimed to investigate the associations between DEHP exposure and serum thyroid hormone levels in 347 adolescents and young adults. We measured DEHP metabolites including mono(2-ethylhexyl) phthalate (MEHP), mono(2-ethyl-5-oxohexyl) phthalate (MEOHP), mono(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP), mono(2-ethyl-5-carboxypentyl) phthalate (MECPP), and mono(2-carboxymethyl)hexyl phthalate (MCMHP) in their urine. Total thyroxine (TT4), total triiodothyronine, free triiodothyronine, free thyroxine (FT4), thyroid-stimulating hormone and the mRNA levels of thyroid peroxidase (TPO), thyroglobulin (TG), sodium iodide symporter (NIS), thyroid transcription factor 1 (TTF-1), and paired box gene 8 (PAX-8) in serum were measured. The results of statistical analysis showed that urinary DEHP metabolites were generally negatively associated with TT4 levels in serum. In the males, the FT4 levels showed positive associations with urinary MEHP, MECPP, MCMHP, and ∑DEHP. The mRNA level of TG was significantly positively correlated with the levels of MECPP, MCMHP, and ∑DEHP, while the levels of TTF-1 and PAX-8 mRNA were significantly positively correlated with the levels of DEHP metabolites. Taken together, DEHP may affect the synthesis of TG by altering the normal transcription of TTF-1 and PAX-8, leading to decreased TT4 levels in Chinese adolescents.


Isolation and characterization of cDNA clones encoding rat skeletal muscle peptidylarginine deiminase.
Various mammalian tissues contain protein-arginine deiminases (EC 22.214.171.124), which convert the arginine residues in normal peptide bonds to the citrulline residues in calcium ion-dependent manners. Here, we describe the complete primary structure of rat skeletal muscle peptidylarginine deiminase deduced from the sequences of its cDNA clones isolated by recombinant DNA technology. We have isolated three overlapping cDNA clones which constitute a 4,507-base pair cDNA sequence including a 2,452-base pair 3'-untranslated region. The coding region consists of 1,995 base pairs encoding 665 amino acid residues. A potential N-linked glycosylation site is present at asparagine-534. The molecular weight of the enzyme calculated from the deduced amino acid sequence is 75,122. Direct repeat sequences resembling the rodent B2 type repetitive sequences appear in the 3'-untranslated region (nucleotides 3,090-3,198 and 3,270-3,391). Northern hybridization demonstrated the presence of its mRNA in poly(A)+ fractions of spinal cord, cerebrum, cerebellum, and submaxillary gland as well as skeletal muscle. The sizes of peptidylarginine deiminase mRNAs in these tissues were estimated to be 4.5-5.0 kilobases. No positive bands were detected on the blots of the corresponding RNA fractions of liver and kidney. Possible similarity of the amino acid sequence of peptidylarginine deiminase to those of other calcium binding proteins is discussed.


Anti-tumour activity of afatinib, an irreversible ErbB family blocker, in human pancreatic tumour cells.
The combination of the reversible epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) erlotinib with gemcitabine obtained FDA approval for treating patients with pancreatic cancer. However, duration of response is often limited and there is currently no reliable predictive marker. We determined the sensitivity of a panel of human pancreatic tumour cell lines to treatment with afatinib, erlotinib, monoclonal antibody (mAb) ICR62, and gemcitabine, using the Sulforhodamine B colorimetric assay. The effect of these agents on cell signalling and cell-cycle distribution was determined by western blot and flow cytometry, respectively. At 200 nM, ICR62 had no effect on growth of these tumour cells with the exception of BxPC-3 cells. BxPC-3 cells were also sensitive to treatment with afatinib and erlotinib with respective IC(50) values of 11 and 1200 nM. Compared with erlotinib, afatinib was also more effective in inhibiting the growth of the other human pancreatic tumour cell lines and in blocking the EGF-induced phosphorylation of tyrosine, EGFR, MAPK, and AKT. When tested in BxPC-3 xenografts, afatinib induced significant delay in tumour growth. The superiority of afatinib in this study encourages further investigation on the therapeutic potential of afatinib as a single agent or in combination with gemcitabine in pancreatic cancer.


Effect of prostaglandin, pyrogen and noradrenaline, injected into the hypothalamus, on thermoregulation in newborn lambs.
Thirteen newborn lambs were implanted stereotaxically with an array of 4 guide tubes positioned bilaterally so that their tips lay above various hypothalamic loci. At postnatal ages 50-70 h, rectal temperature was monitored, and bilateral 1 microliter injections were made into the hypothalamus. Prostaglandin E1 (0.2 microgram) and prostaglandin E2 (0.2-2.0 microgram) did not cause fever on any occasion though injections were made into 44 sites throughout the hypothalamus. Nevertheless, lambs that did not develop fever following central injection were often, if sensitized by bacterial pyrogen given earlier able to develop fever after intravenous injection of bacterial pyrogen. Infusion of a bacterial pyrogen (S. abortus equi, 0.2 microgram) into these same sites caused fever only when injections were made into the anterior hypothalamic/preoptic area (AH/POA). Eight lambs were placed in a 10 degrees C environment and noradrenaline was injected. Rectal temperature fell by 0.5-1.4 degrees C after 7 bilateral injections into the AH/POA, but 9 injections into other hypothalamic areas caused little temperature change. When 6 lambs were placed at a temperature of 30 degrees C and noradrenaline was injected, no consistent temperature changes were observed after microinjection into the AH/POA or other parts of the hypothalamus.


Predicting survival of patients with hypocellular myelodysplastic syndrome: development of a disease-specific prognostic score system.
Although most patients with myelodysplastic syndrome (MDS) exhibit bone marrow hypercellularity, a subset of them present with a hypocellular bone marrow. Specific factors associated with poor prognosis have not been investigated in patients with hypocellular MDS. The authors studied a cohort of 253 patients with hypocellular MDS diagnosed at The University of Texas MD Anderson Cancer Center between 1993 and 2007 and a cohort of 1725 patients with hyper-/normocellular MDS diagnosed during the same time period. Patients with hypocellular MDS presented more frequently with thrombocytopenia (P < .019), neutropenia (P < .001), low serum β-2 microglobulin (P < .001), increased transfusion dependency (P < .001), and intermediate-2/high-risk disease (57% vs 42%, P = .02) compared with patients with hyper-/normocellular MDS. However, no difference in overall survival was observed between the 2 groups (P = .28). Multivariate analysis identified poor performance status (Eastern Cooperative Oncology Group ≥2), low hemoglobin (<10 g/dL), unfavorable cytogenetics (-7/7q or complex), increased bone marrow blasts (≥5%), and high serum lactate dehydrogenase (>600 IU/L) as adverse independent factors for survival. A new prognostic model based on these factors was built that segregated patients into 3 distinct risk categories independent of International Prognostic Scoring System (IPSS) score. This model is independent from the IPSS, further refines IPSS-based prognostication, and may be used to develop of risk-adapted therapeutic approaches for patients with hypocellular MDS.


Relationship between fetal heart rate and umbilical blood flow velocity in term human fetuses during labor.
Fourteen pregnant women between 37 and 41 weeks' gestational age, who subsequently were delivered of healthy newborn infants, were studied to examine the effects of spontaneous changes in fetal heart rate on the flow-velocity waveforms in the umbilical artery. The women were in active labor and under epidural anesthesia at the time Doppler measurements were made. There was a significant positive relationship between end-diastolic flow velocity and fetal heart rate. The umbilical artery peak-systolic/end-diastolic ratio was best correlated with the negative reciprocal transformation of the instantaneous fetal heart rate value and end-diastolic flow velocity. No significant correlation was found between fetal heart rate, umbilical artery peak-systolic/end-diastolic ratio, and peak-systolic flow velocity. An upper limit of 3.0 for a normal umbilical artery peak-systolic/end-diastolic ratio was acceptable only if the instantaneous fetal heart rate was greater than or equal to 130 beats/min.


Multifaceted roles for thymine DNA glycosylase in embryonic development and human carcinogenesis.
Thymine DNA glycosylase (TDG) is a multifunctional protein that plays important roles in DNA repair, DNA demethylation, and transcriptional regulation. These diverse functions make TDG a unique enzyme in embryonic development and carcinogenesis. This review discusses the molecular function of TDG in human cancers and the previously unrecognized value of TDG as a potential target for drug therapy.


SREBP2/Rab11s/GLUT1/6 network regulates proliferation and migration of glioblastoma.
Cholesterol serves a vital role in the occurrence and development of glioblastoma multiforme (GBM). Furthermore, cholesterol synthesis is regulated by sterol regulatory element-binding protein 2 (SREBP2), and certain glucose transporters (GLUTs) and Ras-related protein Rab11 (Rab11) small GTPase family members (Rab11s) may contribute to the process. The Cancer Genome Atlas was used to analyze the relationship between prognosis and GLUT gene expressions. To investigate the regulatory effect of Rab11s and SREBP2 on GLUTs during tumor progression, single cell RNA sequencing (scRNA-seq), western blotting and reverse transcription-quantitative PCR were performed on glioma tissues and the T98G GBM cell line. Cell viability and migration were assessed by performing MTT and wound healing assays, respectively. Moreover, the dual-luciferase reporter gene assay was conducted to predict the sterol regulatory elements in the promoter regions of the target genes. The results demonstrated that high SREBP2, GLUT1 and GLUT6 expression was associated with poor survival of patients with GBM. ScRNA-seq distinguished glioblastoma cells by EGFR and indicated the related lipid metabolism signaling pathways. Moreover, the results indicated that GLUT1 and GLUT6 were regulated by SREBP2 and Rab11s. Rab11s and SREBP2 also contributed to T98G cell viability and migration. Additionally, the results indicated that Rab11s, GLUT1 and GLUT6 were transcriptionally regulated by SREBP2. Therefore, the present study suggested that the SREBP2/Rab11/GLUT network promoted T98G cell growth, thus, identifying potential therapeutic targets for GBM.


Phylosymbiosis in the Rhizosphere Microbiome Extends to Nitrogen Cycle Functional Potential.
Most plants rely on specialized root-associated microbes to obtain essential nitrogen (N), yet not much is known about the evolutionary history of the rhizosphere-plant interaction. We conducted a common garden experiment to investigate the plant root-rhizosphere microbiome association using chloridoid grasses sampled from around the world and grown from seed in a greenhouse. We sought to test whether plants that are more closely related phylogenetically have more similar root bacterial microbiomes than plants that are more distantly related. Using metagenome sequencing, we found that there is a conserved core and a variable rhizosphere bacterial microbiome across the chloridoid grasses. Additionally, phylogenetic distance among the host plant species was correlated with bacterial community composition, suggesting the plant hosts prefer specific bacterial lineages. The functional potential for N utilization across microbiomes fluctuated extensively and mirrored variation in the microbial community composition across host plants. Variation in the bacterial potential for N fixation was strongly affected by the host plants' phylogeny, whereas variation in N recycling, nitrification, and denitrification was unaffected. This study highlights the evolutionary linkage between the N fixation traits of the microbial community and the plant host and suggests that not all functional traits are equally important for plant-microbe associations.


A spontaneous runaway vector for production-scale expression of bovine somatotropin from Escherichia coi.
An Escherichia coli expression vector was constructed for the production-scale fermentation of recombinant bovine somatotropin (rBST). Gene expression is regulated by a spontaneous increase in copy number at a constant low temperature without the need for an external inducer. This vector, designated pURA-4, contains the ampicillin resistance gene, the replication origin from pBR322, the R1 temperature-inducible runaway replicon, and a gene encoding rBST. Optimized rBST expression levels of >35% total cell protein were achieved at a constant 28 degrees C. Shake-flask analysis of pURA-4 shows that the copy number spontaneously increases approximately 6-fold during rBST production. Investigation into the mechanism of pURA-4 spontaneous runaway shows that the increase in copy number is directed by the pBR322 ori and not by the R1 replicon. Although the R1 temperature-inducible replicon does not mediate spontaneous runaway, it does have a positive effect on rBST expression. Copy number analysis also confirmed the stability of pURA-4 spontaneous runaway from the shake-flask scale through the production scale.


The roles and signaling pathways of prolyl-4-hydroxylase 2 in the tumor microenvironment.
Tumor hypoxia is a well-known microenvironmental factor that causes cancer progression and resistance to cancer treatment. Proline hydroxylases (PHDs), a small protein family, belong to an evolutionarily conserved superfamily of dioxygenases, considered the central regulator of the molecular hypoxia response. Prolyl-4-hydroxylase 2 (PHD2), one member of PHDs family, regulates the stability of the hypoxia-inducible factor-1 alpha (HIF-1α) in response to oxygen availability. During hypoxia, the inhibition of PHD2 permits the accumulation of HIF-1α, allowing the cellular adaptation to oxygen limitation, causing activation of numerous genes, which enhances the angiogenesis, metastasis and invasiveness. Accurate regulation of oxygen homeostasis is essential, and which implies PHD2 may have a regulatory role in the pathogenesis of cancer. Although ample evidence exists for a positive correlation between HIFs and tumor formation, metastasis and poor prognosis, the function of the PHD2 in carcinogenesis is less well understood. Despite their original role as the oxygen sensors of the cell and many of the its functions are clearly conveyed through the HIF system, PHD2 is currently known to display HIF-independent and hydroxylase-independent functions in cancer cells and stroma in the control of different cellular pathways. In this review, we summarize the recent advances in the structure, regulation and functions of PHD2 in cancer microenvironment.


Early Prediction of Graft Outcomes After Kidney Transplantation From Donors After Circulatory Death: Biomarkers and Transplantation Characteristics.
This study aimed to identify transplantation characteristics and biomarkers that predict outcomes for kidney transplant (KT) patients from donors after circulatory death (DCDs). Consecutive patients receiving a KT from a DCD in our center between 2014 and 2016 were included; the reference population was recipients with a living donor KT. The urinary tubular injury biomarker-to-creatinine ratio and serum lactate dehydrogenase (LDH) were measured at post-transplant days 1 and 3. The primary outcome was the occurrence of delayed graft function (DGF). Descriptive and receiver operating characteristic analyses were performed. Forty-one patients were included in the analysis: 15 (36.59%) DCD KTs (9 of which suffered from DGF) and 26 (63.41%) living donor KTs. For the primary endpoint, neutrophil gelatinase-associated lipocalin, N-acetyl-beta-D-glucosaminidase, urinary tubular injury biomarker-to-creatinine ratio, and LDH areas under the curve were 1 and 0.96 (95% confidence interval: 0.84-1.0), 1 and 0.92 (95% confidence interval: 0.73-1.0), respectively. Among the transplant characteristics, only the 30-minute resistive index on the perfusion machine was significantly higher in DCD KTs with DGF vs those without DGF (0.26 mm Hg/mL/min [0.20; 0.32] vs 0.14 mm Hg/mL/min [0.12; 0.16], P = .05). Median 3-month creatinine clearance among DGF DCD KTs was 49 mL/min/1.73 m2 [IQR: 42; 65] and 65 mL/min/1.73 m2 [IQR: 62; 66] among DCD KTs without DGF (P = .22). In the DCD KT population, clinical and biological markers were identified that provided predictive tools for DGF. Thus, systematic measurement of these biomarkers, particularly LDH, could improve the management of kidney graft recipients' immunosuppressive therapy.


Antinociception induced in rats by intrathecal administration of antiserum against calcitonin gene-related peptide.
To determine whether or not endogenous calcitonin gene-related peptide (CGRP) participates in pain transmission in the spinal dorsal horn, effects of an intrathecal injection of anti-CGRP antiserum on nociceptive threshold in the paw-pressure test was examined in non-arthritic and adjuvant arthritic rats. An intrathecal injection of the antiserum increased the nociceptive threshold in non-arthritic animals, while 0.9% saline, pre-immune serum and antiserum, previously absorbed by synthetic CGRP, were without effect. Adjuvant arthritic rats showed a hyperalgesia, and improvement occurred with intrathecal injection of the antiserum. Saline and absorbed antiserum were without effect on the hyperalgesia. These results suggest that the endogenous CGRP present in primary afferents probably has a facilitating function in nociceptive transmission in the spinal dorsal horn.


HMOX1 STR polymorphism and malaria: an analysis of a large clinical dataset.
Inducible expression of heme oxygenase-1 (encoded by the gene HMOX1) may determine protection from heme released during malaria infections. A variable length, short tandem GT(n) repeat (STR) in HMOX1 that may influence gene expression has been associated with outcomes of human malaria in some studies. In this study, an analysis of the association between variation at the STR in HMOX1 on severe malaria and severe malaria subtypes is presented in a large, prospectively collected dataset (MalariaGEN). The HMOX1 STR was imputed using a recently developed reference haplotype panel designed for STRs. The STR was classified by total length and split into three alleles based on an observed trimodal distribution of repeat lengths. Logistic regression was used to assess the association between this repeat on cases of severe malaria and severe malaria subtypes (cerebral malaria and severe malarial anaemia). Individual analyses were performed for each MalariaGEN collection site and combined for meta-analysis. One site (Kenya), had detailed clinical metadata, allowing the assessment of the effect of the STR on clinical variables (e.g. parasite count, platelet count) and regression analyses were performed to investigate whether the STR interacted with any clinical variables. Data from 17,960 participants across 11 collection sites were analysed. In logistic regression, there was no strong evidence of association between STR length and severe malaria (Odds Ratio, OR: 0.96, 95% confidence intervals 0.91-1.02 per ten GT(n) repeats), although there did appear to be an association at some sites (e.g., Kenya, OR 0.90, 95% CI 0.82-0.99). There was no evidence of an interaction with any clinical variables. Meta-analysis suggested that increasing HMOX1 STR length is unlikely to be reliably associated with severe malaria. It cannot be ruled out that repeat length may alter risk in specific populations, although whether this is due to chance variation, or true variation due to underlying biology (e.g., gene vs environment interaction) remains unanswered.


Probing the enzyme kinetics, allosteric modulation and activation of α1- and α2-subunit-containing AMP-activated protein kinase (AMPK) heterotrimeric complexes by pharmacological and physiological activators.
AMP-activated protein kinase (AMPK) is a serine/threonine protein kinase that serves as a pleotropic regulator of whole body energy homoeostasis. AMPK exists as a heterotrimeric complex, composed of a catalytic subunit (α) and two regulatory subunits (β and γ), each present as multiple isoforms. In the present study, we compared the enzyme kinetics and allosteric modulation of six recombinant AMPK isoforms, α1β1γ1, α1β2γ1, α1β2γ3, α2β1γ1, α2β2γ1 and α2β2γ3 using known activators, A769662 and AMP. The α1-containing complexes exhibited higher specific activities and lower Km values for a widely used peptide substrate (SAMS) compared with α2-complexes. Surface plasmon resonance (SPR)-based direct binding measurements revealed biphasic binding modes with two distinct equilibrium binding constants for AMP, ADP and ATP across all isoforms tested. The α2-complexes were ∼25-fold more sensitive than α1-complexes to dephosphorylation of a critical threonine on their activation loop (pThr(172/174)). However, α2-complexes were more readily activated by AMP than α1-complexes. Compared with β1-containing heterotrimers, β2-containing AMPK isoforms are less sensitive to activation by A769662, a synthetic activator. These data demonstrate that ligand induced activation of AMPK isoforms may vary significantly based on their AMPK subunit composition. Our studies provide insights for the design of isoform-selective AMPK activators for the treatment of metabolic diseases.


The effect of Paget disease on axillary lymph node metastases and survival in invasive ductal carcinoma.
The objective of this study was to examine the effect of Paget disease (PD) on axillary lymph node metastases and survival in patients who had concomitant invasive ductal carcinoma (PD-IDC). The Surveillance, Epidemiology, and End Results (SEER) database was used to identify women who were diagnosed with PD-IDC from 2000 to 2011, comparing baseline demographic and tumor characteristics with those who were diagnosed with IDC alone during the same period. Multivariable logistic regression was used to examine the association of PD-IDC with axillary lymph node metastasis, and breast cancer-specific survival and overall survival were compared between the PD-IDC and IDC groups using the Kaplan-Meier method and Cox proportional hazards regression. The study cohort included 1102 patients with PD-IDC and 302,242 controls with IDC alone. PD-IDC tumors were more likely to be centrally located (26.9% vs 5.5%; P < .001), high grade (63.5% vs 40.3%; P < .001), >2 cm in greatest dimension (47.1% vs 35.7%; P < .001), and estrogen/progesterone receptor-negative (45.2% vs 22.1%; P < .001). In adjusted analyses, patients with PD-IDC had higher odds of axillary lymph node metastasis (odds ratio, 1.83; P < .001). The unadjusted 10-year breast cancer-specific and overall survival rates were lower for the PD-IDC group compared with the IDC-alone group, although, after adjusting for disease stage, tumor characteristics, and local therapy, no significant differences in mortality risk were observed between the 2 groups (hazard ratio, 0.91; P = .24). PD-IDC is associated with an increased risk of axillary lymph node metastasis, but not with inferior survival, compared with IDC alone after adjustment for other disease factors.


Characterization of the malonyl-CoA-sensitive carnitine palmitoyltransferase (CPTo) of a rat heart mitochondrial particle. Evidence that the catalytic unit is CPTi.
A post 30,000 x g particulate fraction was isolated from rat heart. This mixed membrane fraction is enriched in a carnitine palmitoyltransferase which is sensitive to both malonyl-CoA and etomoxiryl-CoA at concentrations that inhibit the malonyl-CoA-sensitive carnitine palmitoyltransferase (CPTo/CPT-I) of intact mitochondria. Tritiated etomoxiryl-CoA labels two proteins with the same molecular weight as the labeled proteins from rat heart mitochondria. Malonyl-CoA-sensitive carnitine palmitoyltransferase in the particulate fraction is stable to freeze-thawing, and the activity is not latent. These data show that the carnitine palmitoyltransferase associated with this particle is CPTo/CPT-I. Positive Western blots were obtained, with the particle using anti-CPTi/CPT-II at a molecular weight identical with the CPT1/CPT-II purified from rat heart mitochondria. Catalytic activity was purified to near homogeneity in approximately 40% yield. The purified protein has a molecular weight identical with CPTi/CPT-II, it cross-reacts with antibody against CPTi/CPT-II, it is not inhibited by malonyl-CoA or etomoxiryl-CoA, and mass spectral analyses of the tryptic peptides give the same molecular masses as CPTi/CPT-II, and, when mixed with equal amounts of CPTi/CPT-II, one uniform spot is found by two-dimensional electrophoresis. These data indicate that the catalytic subunit of CPTo/CPT-I is the same as CPTi/CPT-II. The average inhibition of the CPT of frozen-thawed particles is 71% by 50 nM etomoxiryl-CoA and 62% by 50 nM malonyl-CoA. The inhibitor sensitivity, but not the catalytic activity, is lost by solubilization in 1% Triton X-114; removal of Triton X-114 using Extracti-Gel D restores etomoxiryl-CoA and malonyl-CoA sensitivity (both 50 nM) of CPT to an average of 77 and 48%, respectively. Consistent with previous reports, these results show that CPTo/CPT-I is NOT inactivated by detergents, rather detergents both desensitize it to malonyl-CoA and alter its Vmax. These data show the assumption that CPTo/CPT-I is inactivated by detergents is untenable.


Suppression by radioactive strontium of the spontaneous cytotoxicity expressed by adherent, predominantly phagocytic cells from various mouse tissues.
Adherent, predominantly phagocytic effector cells which have previously been shown to occur in a large number of tissues of mice and rats have the spontaneous capacity to kill a variety of syngeneic, allogeneic and xenogeneic target cells in vitro. The present work shows that 89Sr treatment of mice suppressed this spontaneous cytotoxicity, thus indicating that the effector cells pass a substantial part of their life in the bone marrow. In contrast, the capacity to mobilize mononuclear phagocytes and to enhance cytotoxicity in response to an appropriate stimulus were not consistently impaired.


Studies on the development of human acute myeloid leukaemia xenografts in immune-deprived mice: comparison with cells in short-term culture.
Human AML cells from the blood of a series of patients have been implanted subcutaneously into mice immune-suppressed by thymectomy and total-body irradiation. Solid tumours resulted from 18 out of 19 samples and their growth was compared with the proliferation of AML cells in culture. In 17 cases tumours grew to a maximum size and then spontaneously regressed. Cells from one patient produced tumours which did not regress and could be retransplanted into freshly immune-suppressed mice. Cells from a human promyelocytic cell line (HL60) also produced nonregressing and retransplantantable tumours. Normal human mononuclear bone marrow cells implanted s.c. produced a growth pattern similar to that of the majority of AML cells. A second inoculum of AML cells into animals with regressing tumours also produced tumours and thus regression cannot be accounted for on the basis of returning immunity. AML cells placed into short-term suspension culture invariably matured to monocyte/macrophage type cells and/or granulocytic cells as identified by cytochemical staining. However, no correlation was observed between proliferation or maturation of cells in culture, and tumour growth in vivo. Cells derived from disaggregated AML tumours also showed evidence of myeloid differentiation suggesting that tumour regression is due to maturation of leukaemic cells.


Oncology drug lag in Japan: has it improved over the last decade?
Existing studies and statistics on the drug lag between Japan and the United States (US) for anti-cancer drugs indicate that it has decreased, whereas more drugs are left unapproved in Japan. This study aimed to quantify the impact of unapproved drugs on the drug lag. Information on 136 anti-cancer drugs approved in the US between 2011 and 2022 was collected. The approval lag, defined as the number of days from the date of approval in the US to the date of approval in Japan, was calculated for all selected drugs, and the median was calculated using the Kaplan-Meier method. The approval lag for drugs not approved in Japan was treated as censored data. Factors potentially associated with the approval lag were explored using Cox regression analysis. The median approval lags for the first half-period (2011-2016) and the last half-period (2017-2022) were 961 days (2.6 years) and 1547 days (4.2 years), respectively (Log-rank test: p = 0.0687). The participation of Japan in the global pivotal trial was associated with a shorter approval lag, and new drug applications by non-Japanese companies that did not rank in the global sales top 20 were associated with a longer approval lag. Drug lag has not decreased over the last decade. The percentage of pivotal trials for US approval that included Japan has increased but should be further increased in the future. Japan may require a scheme to encourage smaller non-Japanese companies to include Japan in their global clinical development plan.


Generation of VHSV replicon particles carrying transmembrane and C-terminal cytoplasmic region-deleted G gene (rVHSV-GΔTM) and comparison of vaccine efficacy with G gene-deleted VHSV (rVHSV-ΔG).
Vaccines based on viral replicon particles would be advantageous to induce immune responses compared to inactivated viruses in that they can infect host cells (only once) and can produce viral proteins in the infected cells like live viruses. Furthermore, as viral replicon particles are replication-defective, they are safer than live attenuated viruses. Previously, we had rescued viral hemorrhagic septicemia virus (VHSV) replicon particles lacking full ORF of G gene (rVHSV-ΔG). In the present study, to enhance the immunogenicity of VHSV replicon particles, we newly generated another form of VHSV replicon particles that can produce the transmembrane and C-terminal cytoplasmic region-deleted G protein in host cells (rVHSV-GΔTM), and compared the protective efficacy of rVHSV-GΔTM with that of rVHSV-ΔG through immunization of olive flounder (Paralichthys olivaceus). In addition, we evaluated the safety of rVHSV-GΔTM by the analysis of effects on wild-type VHSV replication. In the vaccine experiment, olive flounder immunized with rVHSV-GΔTM showed significantly higher titers of serum neutralization activity than fish immunized with rVHSV-ΔG suggesting that the G protein that is not only spiked on the viral envelop but also secreted extracellularly can contribute to the enhancement of adaptive humoral immunity. Moreover, fish immunized with rVHSV-GΔTM showed higher survival rates than fish immunized with rVHSV-ΔG, suggesting that the amount of G protein provided to hosts is an important factor for the enhancement of vaccine efficacy against VHSV disease. In a safety aspect, rVHSV-GΔTM could not replicate in infected cells, and significantly inhibited the replication of wild-type VHSV when co-infected, suggesting that rVHSV-GΔTM would not worsen disease progression caused by wild-type VHSV infection.


T-cadherin in the mammalian cochlea.
Cadherins are a superfamily of transmembrane glycoproteins, which mediate calcium-dependent intercellular adhesions. T-cadherin is an atypical member of the cadherin family in regard to its structure; it acts as a signalling receptor rather than an adhesion molecule. In this study we examine the role of T-cadherin in the mammalian cochlea. This study investigated the expression of T-cadherin in the inner ear under physiologic and pathologic conditions. Expression of T-cadherin in the rat cochlea was analyzed by reverse-transcriptase polymerase chain reaction (RT-PCR), real-time RT-PCR, Western blot, and immunohistochemistry. We detected T-cadherin mRNA expression in three different components in the cochlea of postnatal mouse, namely the organ of Corti (OC), the spiral ganglion (SG), and the stria vascularis (SV). The SG and SV showed a higher T-cadherin mRNA level than the OC. T-cadherin protein was detected by Western blotting in the OC, SG, and SV. Immunofluorescence microscopy of adult mouse cochlea revealed the presence of T-cadherin in the apical parts of the inner and outer hair cells as well as in the SV and SG. OCs treated with gentamicin for 3, 6, or 12 hours did not show any change in T-cadherin gene expression compared to control explants maintained in culture medium alone. T-cadherin is expressed within the cochlea. T-cadherin seems to have a wide variety of functions in the inner ear, ranging from mechanical functions to functions in response to hair cell damage and loss.


Representational cortical plasticity associated with brain tumours: evidence from laser-induced interstitial thermotherapy.
To test the hypothesis that cortical plasticity related to destructive tumour growth is functionally relevant. This hypothesis predicts that function is dependent on the intactness of tissue surrounding the tumour. Eight patients underwent laser-induced interstitial thermotherapy (LITT) for minimally invasive palliative treatment of brain tumours located in eloquent frontal motor regions including the primary motor cortex. A multimodal approach was used to assess the functional outcome of patients after LITT in detail. Following LITT, motor function deteriorated in four patients. In three of these four patients the LITT-induced lesion involved minimal parts of adjacent non-tumorous tissue. By contrast, the other four patients whose LITT-induced signal changes were confined to the tumour, showed no functional deficits. These findings support the idea that peri-tumorous neuronal circuitry in motor competent areas may permanently take over those functions that were formerly represented in the neuronal tissue destroyed by the tumour.


A Segmental Approach in Dorsal Preservation Rhinoplasty: The Tetris Concept.
Rhinoplasty has become a detail-dependent surgery. With precise techniques better results have been achieved. We are continually searching for the ideal technique to offer predictable, accurate, and desired long-term results. Conservative rhinoplasty techniques have been described for more than 100 years as an alternative to dorsal resective surgeries and have reemerged as the new fashion among rhinoplasty surgeons. This article presents our philosophy when approaching the nasal dorsum regarding its segments and their specific anatomic aspects and surgical demands. We describe our intermediate septal strip approach, the intermediate split and the Tetris concept, and discuss their advantages and limitations.


Design, synthesis, in vitro anti-α-glucosidase evaluations, and computational studies of new phthalimide-phenoxy-1,2,3-triazole-N-phenyl (or benzyl) acetamides as potential anti-diabetic agents.
An important target in the treatment of type 2 diabetes is α-glucosidase. Inhibition of this enzyme led to delay in glucose absorption and decrease in postprandial hyperglycemia. A new series of phthalimide-phenoxy-1,2,3-triazole-N-phenyl (or benzyl) acetamides 11a-n were designed based on the reported potent α-glucosidase inhibitors. These compounds were synthesized and screened for their in vitro inhibitory activity against the latter enzyme. The majority of the evaluated compounds displayed high inhibition effects (IC50 values in the range of 45.26 ± 0.03-491.68 ± 0.11 µM) as compared to the positive control acarbose (IC50 value = 750.1 ± 0.23 µM). Among this series, compounds 11j and 11i represented the most potent α-glucosidase inhibitory activities with IC50 values of 45.26 ± 0.03 and 46.25 ± 0.89 µM. Kinetic analysis revealed that the compound 11j is a competitive inhibitor with a Ki of 50.4 µM. Furthermore, the binding interactions of the most potent compounds in α-glucosidase active site were studied through molecular docking and molecular dynamics. The latter studies confirmed the obtained results through in vitro experiments. Furthermore, in silico pharmacokinetic study of the most potent compounds was also performed.


Ferroelectric properties of ultrasonochemically prepared SbSI ethanogel.
This article presents for the first time the electrical properties of sonochemically synthesised, high-surface-area SbSI ethanogel made up of large quantity nanowires with lateral dimensions of about 10-50 nm and lengths reaching up to several micrometers. The composition, morphology, dimensions, microstructures, and optical energy gap of the new form of SbSI were characterized. This material is a semiconducting ferroelectric as in the case of bulk SbSI crystals. The maximum of dielectric constant epsilon=1.6x10(4) is observed at Tc=292(1) K. The activation energies in temperature dependences of electric conductivity of SbSI ethanogel are different for ferroelectric and paraelectric phases during heating and cooling of the sample.


Introduction to Clinical Microbiology for the General Dentist.
Clinical oral microbiology may help dental professionals identify infecting pathogenic species and evaluate their in vitro antimicrobial susceptibility. Saliva, dental plaque biofilms, mucosal smears, abscess aspirates, and soft tissue biopsies are sources of microorganisms for laboratory testing. Microbial-based treatment end points may help clinicians better identify patients in need of additional or altered dental therapies before the onset of clinical treatment failure, and help improve patient oral health outcomes. Microbiological testing appears particularly helpful in periodontal disease treatment planning. Further research and technological advances are likely to increase the availability and clinical utility of microbiological analysis in modern dental practice.


Transcription factor C/EBPα-induced microRNA-30c inactivates Notch1 during granulopoiesis and is downregulated in acute myeloid leukemia.
The transcription factor CCAAT enhancer binding protein α (C/EBPα) is a master regulator in granulopoiesis and is frequently disrupted in acute myeloid leukemia (AML). We have previously shown that C/EBPα exerts its effects by regulating microRNAs (miRs) such as miR-223 and miR-34a. Here, we confirm miR-30c as a novel important target of C/EBPα during granulopoiesis. Thus, wild-type C/EBPα-p42 directly upregulates miR-30c expression, whereas C/EBPα-p30, found in AML, does not. miR-30c is downregulated in AML, especially in normal karyotype AML patients with CEBPA mutations. An induced C/EBPα knockout in mice leads to a significant downregulation of miR-30c expression in bone marrow cells. We identified NOTCH1 as a direct target of miR-30c. Finally, a block of miR-30c prevents C/EBPα-induced downregulation of Notch1 protein and leads to a reduced CD11b expression in myeloid differentiation. Our study presents the first evidence that C/EBPα, miR-30c, and Notch1 together play a critical role in granulocytic differentiation and AML, and particularly in AML with CEBPA mutations. These data reveal the importance of deregulated miRNA expression in leukemia and may provide novel biomarkers and therapeutic targets in AML.