Ichnocarpus frutescens (L.) R. Br. root derived phyto-steroids defends inflammation and algesia by pulling down the pro-inflammatory and nociceptive pain mediators: An in-vitro and in-vivo appraisal.
Ichnocarpus frutescens, a climber plant, is distributed all over India. As its different parts are used as anti-inflammatory agent, so we re-investigated the roots to isolate compounds and evaluate its biological efficacy. Also, in-silico molecular docking was carried out to elucidate the structure activity relationship (SAR) of isolated compounds toward identifies the drug target enzyme with validation, which was further supported by anti-inflammatory in-vitro and in-vivo experimental models. The compounds have been undertaken mainly to investigate the anti-inflammatory and analgesic efficacy along with molecular docking investigation followed by anti-proteinase, anti-denaturation and cyclooxygenase (COX) inhibition studies. Inflammatory cytokines like TNF-α and IL-6 were assayed from lipopolysaccharides (LPS) and Concavallin (CON A) stimulated human PBMC derived macrophages by Enyme linked immune sorbent assay (ELISA) method. The purity index of the lead compound was determined by HPLC. The compounds were illustrated as 2-hydroxy tricosanoic acid (1), stigmasterol glucoside (2), stigmasterol (3), β-sitosterol (4) and β-sitosterol glucoside (5). The test molecules showed significant anti-denaturation, anti-proteinase and analgesic effect validated with docking study. Compounds exhibited anti-inflammatory and pain killing action due to dexamethasone like phytosterol property. Promising anti-denaturation and anti-proteinase activity offered by the compound 5, may hold its promise to fight against arthritis by rejuvenating the osteoblast cells and destroying the bone-resorpting complex of hydrated protein, bone minerals by secreting the acid and an enzyme collagenase along with pain management. The lead bioactive compound i.e. β-sitosterol glucoside (compound 5) demonstrated considerable anti-inflammatory activity showing more than 90% protection against the inflammatory cytokines at 50 µM dose. The anti-denaturation and COX-2 inhibition shown by the compound 5 was also noteworthy with the significant IC50 (ranging from 0.25 to 2.56 µM) that also supporting its future promise for developing as anti-inflammatory agent. Since the most bio-active compound (5) elicit promising acute anti-inflammatory action and peripheral anti-nociceptive pain killing action with a significant ED50 dose of 3.95 & 2.84 mg/kg i.p. respectively in the in-vivo animal model. It could suggest its potentiality as a good in-vivo bio available agent to be an emerging anti-inflammatory drug regimen scaffold in the future. It also establishes significant in-vitro and in-vivo result co-relation. Therefore, the compound 5 could be believed as a potent lead for designing anti-inflammatory, anti-arthritic drug or pain killer without showing any untoward effect.


Plasmodium ovale: in vitro development of hepatic stages.
Primary cultures of human hepatocytes, a culture-derived clone from the human hepatoma Hep G2 line, and cultured rat hepatocytes were inoculated in vitro with Plasmodium ovale sporozoites extracted from Anopheles stephensi, An. gambiae, and An. dirus mosquitoes. Penetration and differentiation of P. ovale sporozoites into trophozoite stage parasites occurred in all three cell types, but with a lower transformation rate in the Hep G2 cell line than in the primary cultured hepatocytes. Further maturation was obtained only in the human hepatocytes, in which the parasites were uninucleate until the third day after infection, before development to 60 micron in length by the eighth day. Additionally, this culture system was used to assess the ability of an anti-P. ovale sporozoite monoclonal antibody to inhibit penetration of sporozoites into hepatocytes and to detect sporozoite determinants in the maturing liver stage parasites.


Differential inhibition produced by peripheral conditioning stimulation on noxious mechanical and thermal responses of different classes of spinal neurons in the cat.
The effect of conditioning stimulation of a peripheral nerve on responses of spinal neurons (dorsal horn cells and motoneurons) was studied in 16 decerebrate-spinal cats. The activity of dorsal horn cells was recorded with a microelectrode at the lumbosacral spinal cord and the single-unit activity of motoneurons was recorded from a filament of ventral rootlet divided from either the L7 or S1 ventral root. The responses of spinal neurons were evoked by noxious and innocuous mechanical stimuli and by noxious thermal stimuli applied to the receptive fields. The peripheral conditioning stimulation was applied to the tibial nerve with repetitive electrical pulses (2 Hz) at an intensity either suprathreshold for A delta or C fibers for 5 min. Applying conditioning stimulation to a peripheral nerve produced a powerful inhibition of the responses elicited by noxious stimuli, suggesting this inhibition is an antinociceptive effect. The inhibition produced by peripheral conditioning stimulation was differentially greater on the responses to noxious than to innocuous stimuli. Based on the results obtained from conditioning stimulation with graded strengths, afferent inputs from both myelinated and unmyelinated fibers seem to contribute to the production of the antinociceptive effect. The magnitude of the antinociceptive effect is bigger for the responses to noxious thermal than to mechanical stimuli. Furthermore, the reflex activity recorded in motor axons seemed to be more sensitive than in dorsal horn cells to the antinociceptive effect.


Endocytoscopy can provide additional diagnostic ability to magnifying chromoendoscopy for colorectal neoplasms.
Pit pattern (PIT) diagnosis with magnifying chromoendoscopy is effective diagnostic method for predicting a massively invasive submucosal colorectal cancer (SMm) which has possibility of metastasis, whereas endocytoscopy (EC) is recently reported to provide excellent diagnostic ability by enabling in vivo cellular visualization. The aim was to assess the additional diagnostic value of EC to PIT for diagnosing colorectal lesions. We conducted a retrospective comparative analysis using a prospectively recorded database in a referral hospital. The subjects were 538 patients who were detected of a colorectal lesion with use of a magnifying colonoscope with EC capability. Each detected lesion was initially diagnosed by PIT findings followed by EC diagnosis by the on-site endoscopist. The diagnostic abilities in predicting neoplastic change and SMm were compared between PIT and PIT plus EC. Overall, 514 lesions from 455 patients were available for analysis. Of them, there were 58 non-neoplastic lesions, 352 adenomas, 15 slightly invasive submucosal cancers, and 89 SMm. The diagnostic abilities of predicting neoplastic change were comparable between PIT and PIT plus EC: sensitivity was 97.8% versus 97.4%, specificity was 91.4% versus 89.7%, and accuracy was 97.1% versus 96.5%. Regarding those of predicting SMm, PIT plus EC showed additional specificity and accuracy to PIT: specificity was 99.1% versus 97.6% (P = 0.041), and accuracy was 96.3% versus 93.8% (P = 0.004). Though PIT has feasible diagnostic ability for predicting both neoplastic change and SMm, EC provides additional diagnostic value to PIT diagnosis for predicting SMm.


Can IgG4-related disease present as isolated myositis?
IgG4-Related Disease (IgG4-RD)is a chronic fibroinflammatory disease typically characterized by inflammation or tumefaction of the organs involved. Skeletal muscle is not one of the typical organs involved in IgG4-RD. Isolated myositis related to IgG4-RD without common organ involvement such as lacrimal or salivary glands or retroperitoneal fibrosis is a controversial and debatable entity. Here we report a case of inflammatory myopathy in an elderly woman with several atypical clinical, lab, and histopathological findings suggestive of IgG4-related myositis. Two such case reports of IgG4-related myositis were reported in the literature review. This is a third case report of elevated IgG4 positive plasma cell infiltration in muscle with severe endomysial fibrosis and unusual myositis features (Figs. 1 and 2). This case-based review opens a possibility of a novel presentation of IgG4-RD and new pathogenesis in myositis.


DNA methylation of RUNX3 promotes the progression of gallbladder cancer through repressing SLC7A11-mediated ferroptosis.
Gallbladder cancer (GBC) is a type of rare but highly aggressive cancer with a dismal prognosis. Runt-related transcription factor 3 (RUNX3), a member of the runt-domain family, and its promoter methylation have been widely observed in a variety of human malignancies. However, the biological function and underlying mechanism of RUNX3 in GBC remain elusive. In this study, bisulfate sequencing PCR (BSP), Western blot, and qPCR were applied to identify the expression level and DNA methylation level of RUNX3 in GBC tissues and cells. The transcriptional relationship between RUNX3 and Inhibitor of growth 1 (ING1) was validated by dual-luciferase reporter assay and ChIP assay. A series of gain-of-function and loss-of-function assays were performed to detect the function and the regulatory relationship of RUNX3 in vitro and in vivo. RUNX3 was aberrantly downregulated in GBC cells and tissues caused by DNA Methyltransferase 1 (DNMT1)-mediated methylation, and downregulation of RUNX3 is associated with poor prognosis of GBC patients. Functional experiments reveal that RUNX3 can induce ferroptosis of GBC cells in vitro and in vivo. Mechanistically, RUNX3 induces ferroptosis by activating ING1 transcription, thereby repressing SLC7A11 in a p53-dependent manner. In conclusion, the downregulation of RUNX3 is mediated by DNA methylation, which promotes the pathogenesis of gallbladder cancer through attenuating SLC7A11-mediated ferroptosis. This study gives novel insights into the role of RUNX3 in the ferroptosis of GBC cells, which may contribute to developing potential treatment targets for GBC.


Cell migration regulated by RGD nanospacing and enhanced under moderate cell adhesion on biomaterials.
While nanoscale modification of a biomaterial surface is known to influence various cell behaviors, it is unclear whether there is an optimal nanospacing of a bioactive ligand with respect to cell migration. Herein, we investigated the effects of nanospacing of arginine-glycine-aspartate (RGD) peptide on cell migration and its relation to cell adhesion. To this end, we prepared RGD nanopatterns with varied nanospacings (31-125 nm) against the nonfouling background of poly(ethylene glycol), and employed human umbilical vein endothelial cells (HUVECs) to examine cell behaviors on the nanopatterned surfaces. While HUVECs adhered well on surfaces of RGD nanospacing less than 70 nm and exhibited a monotonic decrease of adhesion with the increase of RGD nanospacing, cell migration exhibited a nonmonotonic change with the ligand nanospacing: the maximum migration velocity was observed around 90 nm of nanospacing, and slow or very slow migration occurred in the cases of small or large RGD nanospacings. Therefore, moderate cell adhesion is beneficial for fast cell migration. Further molecular biology studies revealed that attenuated cell adhesion and activated dynamic actin rearrangement accounted for the promotion of cell migration, and the genes of small G proteins such as Cdc42 were upregulated correspondingly. The present study sheds new light on cell migration and its relation to cell adhesion, and paves a way for designing biomaterials for applications in regenerative medicine.


Phace Syndrome in Children: Two Case Reports.
PHACE(S) syndrome combines: posterior fossa brain malformations, face hemangioma, arterial cerebrovascular abnormalities, cardiovascular abnormalities, eye abnormalities, and ventral developmental defects (Sternal defects or supra-umbilical rope). The diagnosis is based on the association of an infant hemangioma exceeding 5cm in size on the face, neck, scalp with 1 major criteria or 2 minor criteria. Imaging, especially Gadolinium MRI and MRA of the brain, neck, and aortic arch, transthoracic echocardiography, or even cardiac MRI play a key role in the detection of associated neurological and cardiovascular abnormalities. We report 2 cases of PHACE syndrome revealed by 2 different clinical presentations.


Resistance of oat breeding lines to grain contamination with Fusarium langsethiae and T-2/HT-2 toxins.
Fusarium disease of oats reduces yield quality due to decreasing germination that is caused by the contamination of grain with mycotoxins produced by Fusarium fungi. The aim of this study was to characterize the resistance of naked breeding lines of oats to fungal grain infection and to contamination with T-2 and HT-2 toxins. Thirteen naked oat breeding lines and two naked varieties, Nemchinovsky 61 and Vyatskiy, as well as a husked variety Yakov, were grown under natural conditions in the Nemchinovka Federal Research Center in 2019-2020. The contamination of grain with fungi was determined by the mycological method and real-time PCR. The analysis of mycotoxins was carried out by ELISA. In oats, Alternaria (the grain infection was 15-90 %), Cochliobolus (1-33 %), Cladosporium (1-19 %), Epicoccum (0-11 %), and Fusarium (3-17 %) fungi prevailed in the grain mycobiota. The predominant Fusarium species were F. poae (its proportion among Fusarium fungi was 49-68 %) and F. langsethiae (29-28 %). The highest amounts of F. langsethiae DNA ((27.9-71.9) × 10-4 pg/ng) and T-2/HT-2 toxins (790-1230 μg/kg) were found in the grain of husked oat Yakov. Among the analysed naked oat lines, the amount of F. langsethiae DNA varied in the range of (1.2-42.7) × 10-4 pg/ng,and the content of T-2/HT-2 toxins was in the range of 5-229 μg/kg. Two oat breeding lines, 54h2476 and 66h2618, as well as a new variety, Azil (57h2396), can be characterized as highly resistant to infection with Fusarium fungi and contamination with mycotoxins compared to the control variety Vyatskiy.


Crystal structure of ethyl 2-{4-[(2-oxo-3-phenyl-1,2-di-hydro-quinoxalin-1-yl)meth-yl]-1H-1,2,3-triazol-1-yl}acetate.
The quinoxaline portion of the title mol-ecule, C21H19N5O3, is not quite planar as indicated by a dihedral angle of 3.38 (7)° between the constituent rings. The mol-ecule is 'U-shaped', which is consolidated by an intra-molecular anti-parallel carbonyl electrostatic inter-action with C··O distances of 2.8905 (16) and 3.0221 (15) Å, in the crystal forms corrugated layers through C-H⋯O and C-H⋯N hydrogen bonds and C-H⋯π(ring) and π-stacking inter-actions.


Monitoring of functional residual capacity by an oxygen washin/washout; technical description and evaluation.
It was the goal of this study to develop and test an automated method for measuring functional residual capacity (FRC) by an oxygen washin/washout in intensive care settings. Such a method is required to work with conventional ventilator breathing systems and to use only medical grade sensors. The oxygen setting on a standard intensive care ventilator is changed by at least 10%. Ventilatory pressure and flow are measured by the built-in sensors of the intensive care ventilator. Oxygen concentration is measured by a diverting medical oxygen analyzer. In order to overcome the known problem that synchrony between flow and concentration measurement is corrupted by the change of gas viscosity and by the cyclic change of airway pressure, a physical/mathematical model of the pneumatic circuit of the analyzer was developed. With this model, the change of sample flow is calculated continuously. Thus, synchrony between flow and gas concentration measurement is restored. This allows the determination of volumetric gas fluxes as needed for the FRC measurement. The setup was tested in the laboratory with a lung simulator. Simulated lung compliance, breathing frequency and tidal volume were varied. Results. The mean difference between measured and simulated FRC (range 1.7 to 5 L) was less than 1% at tidal volumes greater than 400 mL. This difference ranged from -5% to 8%, depending on simulated lung compliance and ventilator setting. The variability of consecutive measurements was about 2.5%. A method has been developed for reliable measurement of the FRC with an oxygen washin/washout technique. This method is sufficiently easy to use to suit for application in intensive care units. It does not require any action by the operator except a manual change of inspired oxygen concentration. Accuracy and sensitivity of the method have been proven sufficient to meet clinical and scientific requirements. Future clinical studies will reveal the applicability of the chosen procedure under clinical conditions.


Selective embryo reduction in a sextuplet pregnancy.
The treatment course of a 29-year-old infertile woman using hMG and TDI was the cause of a sextuplet pregnancy in which a successful embryo reduction was carried out during the first trimester. The reduction was performed twice: the first around the 9th week of pregnancy transvaginally, and the second during the 12th week by transabdominal puncture. The perinatal outcome was favorable, and the patient spontaneously delivered a healthy infant on the 39th week of pregnancy.


Identification of the active compounds in the Yi-Fei-San-Jie formula using a comprehensive strategy based on cell extraction/UPLC-MS/MS, network pharmacology, and molecular biology techniques.
Chinese herbal formulae has multiple active constituents and targets, and the good clinical response is encouraging more scientists to explore the bio-active ingredients in such complex systems. Yi-Fei-San-Jie formula (YFSJF) is commonly used to treat patients with lung cancer in South China; however, its bio-active ingredients remain unknown. We investigated the bio-active ingredients of the YFSJF using a novel comprehensive strategy. A549 cell extraction coupled with ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS/MS) was used for the screening of potential bio-active ingredients. Network pharmacology approach and molecular dynamics simulation were performed for the screening of targets. Surface plasmon resonance (SPR) assay and molecular biology techniques were used to verify the targets. Nine A549 cell membrane-binding compounds were identified through cell extraction/UPLC-MS/MS. Five compounds, namely ginsenoside Ro, ginsenoside Rb1, ginsenoside Rc, peimisine, and peimine were cytotoxic to A549 cells, and they were considered the bio-active ingredients of the YFSJF in vitro. Network pharmacology analysis revealed that TGFBR2 is the key target and the TGFβ pathway is the key pathway targeted by YFSJF in non-small cell lung cancer. Peimisine showed an affinity to TGFBR2 using molecular docking and dynamic stimulation, which was confirmed using surface plasmon resonance spectroscopy. The molecular biology-based analysis further confirmed that peimisine targets TGFBR2 and can reverse A549 epithelial-mesenchymal transition by inhibiting the TGFβ pathway. Taken together, cell extraction/UPLC-MS/MS, network pharmacology, and molecular biology-based analysis comprise a feasible strategy to explore active ingredients in YFSJF.


Novel spiroindoline quinazolinedione derivatives as anticancer agents and potential FLT3 kinase inhibitors.
Despite considerable recent progress in therapeutic strategies, cancer still remains one of the leading causes of death. Molecularly targeted therapies, in particular those focused on blocking receptor tyrosine kinases have produced promising outcomes in recent years. In this study, a new series of spiro[indoline-3,2'-quinazoline]-2,4'(3'H)-dione derivatives (5a-5l) were synthesized and evaluated as potential kinase inhibitors with anticancereffects. The anti-proliferative activity was measured by MTT assay, while the cell cycle was studied using flow cytometry. Moreover, kinase inhibition profiles of the most promising compounds were assessed against a panel of 25 oncogenic kinases. Compounds 5f,5g,5i, and 5jshowed anti-proliferative effect against EBC-1, A549, and HT-29 solid tumor models in addition to leukemia cell line K562. In particular, compound 5f, bearing 4-methylphenyl pendant on the isatin ring displayed considerable potency with IC50 values of 2.4 to 13.4 μM against cancer cells. The most potent derivatives also altered the distribution of cells in different phases of cell cycle and increased the sub-G1 phase cells in K562 cells. Moreover, kinase inhibition assays identified FLT3 kinase was as the primary targetof these derivatives. Compound 5f at 25 μM concentration showed inhibitory activities of 55% and 62% against wild-type FLT3 and its mutant, D835Y, respectively. Finally, the docking and simulation studies revealed the important interactions of compound 5f with wild type and mutant FLT3. The results of this study showed that some novel spiroindoline quinazolinedione compounds could be potential candidates for further development as novel targeted anticancer agents.


Inhibitory effect of anti-obesity drugs on NADH dehydrogenase of mouse heart homogenates.
Propylhexedrine and chlorphentermine inhibit the activity of NADH cytochrome c reductase but to a lesser extent than fenfluramine. Diethylpropion and phendimetrazine have little effect.


Impact of CYP2D6 genotype on adverse effects during treatment with metoprolol: a prospective clinical study.
Our objective was to study the impact of the cytochrome P450 (CYP) 2D6 polymorphism on the tolerability of metoprolol in a real-life primary care setting. The adverse effects studied comprised effects related to the central nervous system, cardiovascular effects, and sexual dysfunction. Patients in whom treatment with metoprolol was considered were enrolled into this prospective, 6-week multicenter study. The dosage of metoprolol was determined on an individual basis and could be freely adjusted on clinical grounds. The indication for treatment was hypertension in about 90% of cases. Systolic and diastolic blood pressure, resting heart rate, and plasma metoprolol and alpha-hydroxymetoprolol concentrations were measured. CYP2D6 genotyping covered alleles *3 to *10 and *41 and the duplications. Possible adverse effects of metoprolol were systematically assessed over a 6-week period by means of standardized rating scales and questionnaires. The final study population comprised 121 evaluable patients (all white patients); among them, there were 5 ultrarapid metabolizers (UMs) (4.1%), 91 extensive metabolizers (EMs) (75%), 21 intermediate metabolizers (IMs) (17%), and 4 poor metabolizers (PMs) (3.3%). Plasma metoprolol concentrations normalized for the daily dose and metoprolol/alpha-hydroxymetoprolol ratios at steady state were markedly influenced by CYP2D6 genotype and displayed a gene-dose effect. The median of the dose-normalized metoprolol concentration was 0.0088 ng/mL, 0.047 ng/mL, 0.34 ng/mL, and 1.34 ng/mL among UMs, EMs, IMs, and PMs, respectively (P<.0001). There was no significant association between CYP2D6 genotype-derived phenotype (EMs and UMs combined versus PMs and IMs combined) and adverse effects during treatment with metoprolol. There was a tendency toward a more frequent occurrence of cold extremities in the PM plus IM group as compared with the EM plus UM group (16.0% versus 4.2%, P=.056; relative risk, 3.8 [95% confidence interval, 1.03--14.3]). CYP2D6 genotype-derived phenotype was not significantly associated with a propensity for adverse effects to develop during treatment with metoprolol. However, the results concerning tolerability of metoprolol in PMs were inconclusive because of the small number of PMs enrolled.


Evidence of Brucella abortus OPS dictating uptake and restricting NF-kappaB activation in murine macrophages.
Smooth Brucella abortus S2308 is virulent while rough derivatives are attenuated. Intracellular killing is often blamed for these differences. In the studies described, uptake kinetics and interaction of S2308 and S2308 manBA::Tn5 (CA180) rough mutants with macrophages were investigated. The results revealed that smooth B. abortus was rapidly internalized, achieving a maximum level in less than 5 min without additional uptake over the next 30 min. In contrast, continued uptake of the rough mutant was observed and only achieves a maximum level after 30 min. The results were confirmed by the differences in F-actin polymerization, lipid raft staining, early endosome colocalization and electron microscopic observations after smooth and rough Brucella infection. We also demonstrated for the first time that uptake of S2308, but not rough mutant CA180 was PI3-kinase and toll-like receptor 4 (TLR4) dependent. Differences in uptake were associated with differences in macrophage activation with regard to NF-kappaB translocation and cytokine production. These results provide evidence that the presence of B. abortus OPS dictates the interactions between Brucella and specific cell surface receptors minimizing macrophage activation and enhancing Brucella survival and/or persistence.


Tissue-specific transcriptome for Dendrobium officinale reveals genes involved in flavonoid biosynthesis.
Dendrobium officinale is a kind of crop and precious herbal that is widely distributed in China. We applied transcriptomics to investigate the flavonoids and their biosynthesis-related genes from different tissues. Total flavonoid was determined in three different tissues. In this study, nine cDNA libraries were generated from Dendrobium officinale. A total of 530 million (70.73%) of the high-quality reads were successfully mapped to the reference genome of Dendrobium officinale and nine libraries were combined and assembled into 24,927 non-redundant genes. We mapped all of these genes to reference pathways in the KEGG database to identify polysaccharide and secondary metabolites pathways in which the genes may be involved. We outlined the biosynthetic pathway of flavonoids and identified putative genes, which provide understanding of the biosynthesis and regulation of Dendrobium officinale flavonoids at the molecular level. We identified five CHS genes from Dendrobium officinale and characterized the CHS gene family. The flavonoid-related key enzyme genes were identified, and their expression patterns in different tissues were further analyzed using quantitative real-time PCR. These data on flavonoid genes obtained in this work will be useful in understanding the molecular mechanisms of different tissues in Dendrobium officinale.


Outpatient catheterless modified Mathieu hypospadias repair.
Distal penile hypospadias was repaired in 59 boys with a modification of the Mathieu technique. Since a watertight urethroplasty is performed without overlying suture lines, catheter drainage and urinary diversion may not be necessary and they were not used in any patient. Penile nerve block and a small dressing allowed for postoperative mobilization. An excellent cosmetic and functional result was achieved with few complications, even in boys who had had prior unsatisfactory repairs. Because the children are ambulatory postoperatively and tolerate voiding through the repair, this procedure often is performed on an outpatient basis.


Baccharis trimera: effect on hematological and biochemical parameters and hepatorenal evaluation in pregnant rats.
This investigation evaluated the effect of a hydroethanolic extract of Baccharis trimera on pregnant Wistar rats, once the plant is well-known consumed in pregnancy and little is known on its potentially toxic effects on pregnant women. Thirty-five female rats were distributed into three groups. Those in treatments 1 and 2 were given 8.4 mg/kg of the extract orally from gestational day (GD) 1 to 19 and from GD 6 to 15, respectively, whereas those in the control group received distilled water orally from GD 1 to 19. Body weights were recorded on GD 1, 6, 15, and 20. On GD 20 animals were anesthetized, blood samples were collected and maternal livers, kidneys, and spleens were weighed and processed for histological studies. No clinical signs of maternal toxicity and no changes in hematological parameters were observed. Urea levels and kidney weights differed significantly between animals receiving treatment 1 and controls. Histopathological alterations were found in kidneys and livers in both treatment groups. The hydroethanolic extract of Baccharis trimera administered to pregnant rats at 8.4 mg/kg was toxic to maternal kidney and liver cells, although such alterations are reversible once administration is discontinued.


The effects of flumazenil-precipitated abstinence on the pharmacokinetics of chronic oxazepam in dogs.
The pharmacokinetics of oxazepam was studied in naive dogs and in oxazepam-dependent dogs without and with administered flumazenil (6 mg/kg). Oxazepam is eliminated with a relatively short elimination half life (ca. 150 min) in both acutely and chronically treated dogs. It exhibits only a modest first pass metabolism (ca. 10%) and its bioavailability following oral administration is about 22%. The steady state concentration of oxazepam in chronically treated dogs was lower than was predicted from single dose studies. Flumazenil did not change the rate of absorption or elimination of oxazepam-dependent dogs. The total steady state plasma concentration of oxazepam was significantly reduced by flumazenil administration suggesting a displacement interaction between flumazenil and oxazepam.


Identification of recurring patterns in fractionated atrial electrograms using new transform coefficients.
Identification of recurrent patterns in complex fractionated atrial electrograms (CFAE) has been used to differentiate paroxysmal from persistent atrial fibrillation (AF). Detection of the atrial CFAE patterns might therefore be assistive in guiding radiofrequency catheter ablation to drivers of the arrhythmia. In this study a technique for robust detection and classification of recurrent CFAE patterns is described. CFAE were obtained from the four pulmonary vein ostia, and from the anterior and posterior left atrium, in 10 patients with paroxysmal AF and 10 patients with longstanding persistent AF (216 recordings in total). Sequences 8.4 s in length were analyzed (8,192 sample points, 977 Hz sampling). Among the 216 sequences, two recurrent patterns A and B were substituted for 4 and 5 of the sequences, respectively. To this data, random interference, and random interference + noise were separately added. Basis vectors were constructed using a new transform that is derived from ensemble averaging. Patterns A and B were then detected and classified using a threshold level of Euclidean distance between spectral signatures as constructed with transform coefficients. In the presence of interference, sensitivity to detect and distinguish two patterns A and B was 96.2%, while specificity to exclude nonpatterns was 98.0%. In the presence of interference + noise, sensitivity was 89.1% while specificity was 97.0%. Transform coefficients computed from ensemble averages can be used to succinctly quantify synchronized patterns present in AF data. The technique is useful to automatically detect recurrent patterns in CFAE that are embedded in interference without user bias. This quantitation can be implemented in real-time to map the AF substrate prior to and during catheter ablation.


Optimization of LC-MS/MS using triple quadrupole mass analyzer for the simultaneous analysis of carbosulfan and its main metabolites in oranges.
This paper describes an analytical method involving a simple solvent extraction for the simultaneous liquid chromatography coupled to quadrupole tandem mass spectrometry (LC-MS/MS) determination of carbosulfan, its most toxic metabolite--carbofuran--, and its other main metabolites--3-hydroxycarbofuran, 3-ketocarbofuran, 3-hydroxy-7-phenolcarbofuran, 3-keto-7-phenolcarbofuran, 7-phenolcarbofuran and dibutylamine--in oranges. Chromatography was performed on a Zorbax Bonus-RP (150 mm x 2.1 mm, 5 microm). The mobile phase was a ternary gradient water-methanol-acetonitrile with 1.0 mM ammonium acetate at flow rate of 0.2 ml min(-1). The LC separation and MS/MS optimization were studied to select the most appropriate operating conditions. The method developed has also been validated. The limits of quantification (LOQs) were from 1 microg kg(-1) for carbofuran to 10 microg kg(-1) for 3-keto-7-phenolcarbofuran. Extracts spiked with carbosulfan and its metabolites, at LOQ level, yielded average recoveries in the range 60-94%, with relative standard deviations (R.S.D.s) less than 15%. Calibration curves for carbosulfan and its metabolites (range LOQ-1000LOQ) were linear, with coefficients of correlations better than 0.990. The method was successfully applied to establish the primary degradation products in oranges treated with carbosulfan. The LC-MS/MS method developed is simple, rapid, and suitable for the quantification and confirmation of carbosulfan and seven of its main metabolites in orange at levels lower than 10 microg kg(-1).


Gender proteomics I. Which proteins in non-sexual organs.
Differences related to gender have long been neglected but recent investigations show that they are widespread and may be recognized with all types of omics approaches, both in tissues and in biological fluids. Our review compiles evidence collected with proteomics techniques in our species, mainly focusing on baseline parameters in non-sexual organs in healthy men and women. Data from human specimens had to be replaced with information from other mammals every time invasive procedures of sample procurement were involved. As our knowledge, and the methods to build it, get refined, gender differences need to receive more and more attention, as they influence the outcome of all aspects in lifestyle, including diet, exercise and environmental factors. In turn this background modulates a differential susceptibility to some disease, or a different pathogenetic mechanism, depending on gender, and a different response to pharmacological therapy. Preparing this review we meant to raise awareness about the gender issue. We anticipate that more and more often, in the future, separate evaluations will be carried out on male and female subjects as an alternative - and an upgrade - to the current approach of reference and test groups being 'matched for age and sex'.


Strategies for attraction and retention of health workers in remote and difficult-to-access areas of Chhattisgarh, India: Do they work?
To address the acute shortages of health workers in underserved, remote, and difficult-to-access areas, the Government of Chhattisgarh and the National Rural Health Mission (NRHM) launched the Chhattisgarh Rural Medical Corps (CRMC) in 2009. CRMC has enabled provisions such as financial incentives, residential accommodation, life insurance, and extra marks during admission at the postgraduate (PG) level to eligible doctors for the attraction and retention of health workers, i.e., doctors, staff nurses, auxiliary nurse midwives (ANMs), and rural medical assistants (RMAs) in underserved areas. This study aims to understand the CRMC scheme in terms of implementation, challenges, gaps, and outcome in achieving the attraction and retention of health workers in the remote and difficult-to-access areas of Chhattisgarh. The study adopts a mix of both qualitative and quantitative research methods. The purposive sampling method was used for the selection of three districts having normal, difficult, and inaccessible areas. Data were collected through key informant (KI) interviews with beneficiaries and non-beneficiaries of CRMC or district and state government officials, and reviews of document were analyzed using a thematic analysis approach. CRMC has made positive outcome as 1319 health workers, including doctors, have joined the service in 2010-11, reducing the vacancy of doctors from 90% to 45%. The scope of CRMC was primarily limited to payment of monthly financial incentives. The fund utilization rate of CRMC has increased (from 27% in 2009-10 to 98% in 2011-12), though there are delays in payment of incentives. The majority of staff lack awareness about CRMC during job applications. The payment of incentives based on facility performance has demotivated staff. Establishment of a performance management system, activating the CRMC cell to make it functional, and wide publicity of CRMC benefits are likely to improve attraction and retention of staff.


The influence of (+)-propranolol on the inotropic effect of dihydro-ouabain in relation to stimulation frequency.
1. The influence of (+)-propranolol, 10(-6) to 4 x 10(-5) mol/l, upon the positive inotropic effect of dihydro-ouabain was studied on guinea-pig papillary muscle. At 1 Hz, negative inotropically effective concentrations of (+)-propranolol decreased the inotropic effectiveness of the glycoside. 2. For equivalent diminution of the basal force of contraction, either by reduction of stimulation frequency or by increasing concentrations of (+)-propranolol, a similar decrease in the inotropic effectiveness of dihydro-ouabain was observed. In contract, reduction of [Ca2+]o had almost no effect on the effectiveness of the glycoside. 3. The inhibitory effect of (+)-propranolol on the inotropic effectiveness of dihydro-ouabain declined with reduction of stimulation frequency. At any stimulation frequency tested, the inhibitory effect of (+)-propranolol was proportional to the effect of a reduction of that particular frequency to the rested-state condition in the absence of (+)-propranolol. 4. The results are consistent with the hypothesis that the inhibition of the excitation-coupled sodium influx by (+)-propranolol is the cause of its inhibiting effect on the effectiveness of dihydro-ouabain.


Use of 18F-FDG PET-CT imaging to determine internal mammary lymph node location for radiation therapy treatment planning in breast cancer patients.
Adjuvant internal mammary lymph node (IMN) radiation is often delivered with 2-dimensional techniques that use anatomic landmarks and predetermined depths for field placement and dose specification. In contrast, 3-dimensional planning uses the internal mammary vessels (IMVs) to localize the IMNs for planning. Our purpose was to determine if localization of the involved IMN (i-IMN) by 18F-labeled fluorodeoxyglucose positron emission tomography-computed tomography (18F-FDG PET-CT) offers opportunities to improve treatment. Breast cancer patients (n = 80) who had i-IMNs (n = 112) on PET-CT for initial staging (n = 40) or recurrence (n = 40) were studied. Size, intercostal space (IC), and distance from skin, sternum, and IMVs were recorded. Effects on 2- and 3-dimensional planning were evaluated. Most i-IMNs (94.6%) were in the first to third ICs. Few were in the fourth (4.5%) or fifth (0.9%) IC. Mean i-IMN depth was 3.4 cm (range, 1.1-7.3 cm). Prescriptive depths of 4, 5, and 6 cm would result in undertreatment of 25%, 10.7%, and 5.3% of IMNs, respectively. Most IMNs (86.6%) were lateral or adjacent to the sternal edge. Only 13.4% of IMNs were posterior to the sternum. Use of the ipsilateral or contralateral sternal edge for field placement increases the risk of geographic miss or excess normal tissue exposure. Most i-IMNs were adjacent to (83%) or ≤0.5 cm (14%) from the IMV edge. Three (3%) were >0.5 cm beyond the IMV edge. The clinical target volume (CTV) defined by the first to third ICs encompassed 78% of i-IMNs. IMN-CTV coverage of i-IMNs increased with inclusion of the fourth IC (82%), 0.5 cm medial and lateral margin expansion (93%), or both (96.5%). Two-dimensional treatment techniques risk geographic miss of IMNs and exposure of excess normal tissue to radiation. An IMN-CTV defined by the IMVs from the first to third ICs with 0.5-cm medial and lateral margin expansion encompasses almost all i-IMNs identified on PET-CT imaging. Inclusion of the fourth IC offers modest coverage improvement, and its inclusion should be weighed against potential increase in cardiac exposure. The use of 2-dimensional treatment techniques for adjuvant internal mammary lymph node (IMN) radiation may cause geographic miss of tumor and expose normal tissue to radiation injury. Conformal 3-dimensional planning improves coverage and reduces risk of normal tissue damage by using the internal mammary vessel to define an IMN clinical target volume (CTV). Contouring the IMN-CTV from the first to third intercostal spaces with a 0.5-cm expansion medially and laterally encompasses most IMN. Positron emission tomography-computed tomography may have a role in radiation planning by identifying involved-IMN for dose escalation.


Effects of hip joint angle changes on intersegmental spinal coupling in human spinal cord injury.
Pathological expression of movement and muscle tone in human upper motor neuron disorders has been partly associated with impaired modulation of spinal inhibitory mechanisms, such as reciprocal or presynaptic inhibition. In addition, input from specific afferent systems contributes significantly to spinal reflex circuits coupled with posture or locomotion. Accordingly, the objectives of this study were to identify the involved afferents and their relative contribution to soleus H-reflex modulation induced by changes in hip position, and to relate these effects with activity of spinal interneuronal circuits. Specifically, we investigated the actions of group I synergistic and antagonistic muscle afferents (e.g. common peroneal nerve, CPN; medial gastrocnemius, MG) and tactile plantar cutaneous afferents on the soleus H-reflex during controlled hip angle variations in 11 motor incomplete spinal cord injured (SCI) subjects. It has been postulated in healthy subjects that CPN stimulation evokes an inhibition on the soleus H-reflex at a conditioning test (C-T) interval of 2-4 ms. This short latency reflex depression is caused mainly by activation of the reciprocal Ia inhibitory pathway. At longer C-T intervals (beyond 30 ms) the soleus H-reflex is again depressed, and is generally accepted to be caused by presynaptic inhibition of soleus Ia afferents. Similarly, MG nerve stimulation depresses soleus H-reflex excitability at the C-T interval of 6 ms, involving the pathway of non-reciprocal group I inhibition, while excitation of plantar cutaneous afferents affects the activity of spinal reflex pathways in the extensors. In this study, soleus H-reflexes recorded alone or during CPN stimulation at either short (2, 3, 4 ms) or long (80, 100, 120 ms) C-T intervals, and MG nerve stimulation delivered at 6 ms were elicited via conventional methods and similar to those adopted in studies conducted in healthy subjects. Plantar skin conditioning stimulation was delivered through two surface electrodes placed on the metatarsals at different C-T intervals ranging from 3 to 90 ms. CPN stimulation at either short or long C-T intervals and MG nerve stimulation resulted in a significant facilitation of the soleus H-reflex, regardless of the hip angle tested. Plantar skin stimulation delivered with hip extended at 10 degrees induced a bimodal facilitation reflex pattern, while with hip flexed (10 degrees , 30 degrees ) the reflex facilitation increased with increments in the C-T interval. This study provides evidence that in human chronic SCI, classically key inhibitory reflex actions are switched to facilitatory, and that spinal processing of plantar cutaneous sensory input and actions of synergistic/antagonistic muscle afferents interact with hip proprioceptive input to facilitate soleus H-reflex excitability. These actions might be associated with the pathological expression of neural control of movement in individuals with SCI, and potentially could be considered in rehabilitation programs geared to restore sensorimotor function in these patients.


Cloudy outlook. Supercommittee failure leaves healthcare providers questioning future cuts, impact on hospitals.
President Barack Obama responded to the failure of the so-called supercommittee with a message that he won't allow Congress to water down the automatic cuts triggered under the August deficit law, which would include a 2% reduction in Medicare payments.


Polypeptide patterns of infectious bronchitis virus serotypes fall into two categories. Brief report.
Molecular weights of six major polypeptides of infectious bronchitis virus (IBV) are: 1. 75,000; 2. 50,000; 3. 45,000; 4. 35,000; 5. 28,000 or 24,000, and 6. 22,000 dalton. According to the mobility of protein 5 the polypeptide patterns of IBV serotypes fall into two main categories.


Differentiating clinical and non-clinical depression: a heuristic study offering a template for extension studies.
To differentiate clinical and non-clinical depression via a set of symptoms. A sample of 140 patients attending a clinical service for those with mood disorders together with 40 subjects denying ever experiencing a clinical episode of depression were compared, with participants completing a questionnaire capturing many symptoms of depression as well as illness correlates. A latent class analysis of symptom data identified two classes and with class assignment corresponding strongly with initial clinical vs. non-clinical assignment. Univariate analyses identified the extent to which individual symptoms contributed to differentiation. Study data suggested DSM criteria that would benefit from re-writing or of reassignment. Two models for classifying clinical depression were generated. The first involved individuals feeling hopeless and also being suicidal or at risk of self-harm. The second involved a symptom set corresponding to DSM-5 criteria but with only five making significant independent contributions to diagnostic differentiation. The study is heuristic in offering a strategy for more precisely differentiating clinical and non-clinical depression in more representative samples, so allowing resolution of key features, and determining whether a monothetic or polythetic diagnostic symptom criterion model is optimal.


Development and evaluation of a chemiluminescence immunoassay for detecting tropical theileriosis.
Tropical theileriosis is a tick-borne lymphoproliferative disease of cattle caused by the apicomplexan parasite Theileria annulata, and leads to substantial economic losses to the livestock industry worldwide. Although various enzyme-linked immunosorbent assays (ELISAs) have been established to detect antibodies against T. annulata infection, a specific, rapid and reliable diagnostic assay is urgently needed for prevention and control of the disease. In the present study, a chemiluminescence immunoassay (CLIA) was developed based on the subtelomeric variable secreted protein (SVSP) of T. annulata as a sero-diagnostic antigen. Following optimization of the CLIA working parameters, the working time of the method was less than 4.5 h. The sensitivity and specificity of the established CLIA was 98.8% and 97.5%, respectively, when the cut-off value of the percent positive (PP) was 26.1% for detecting serum samples (n = 242 T. annulata positive sera, n = 158 T. annulata negative sera). After comparing 180 serum samples from Gansu province, China, the concordance rate between the CLIA and a published rSpm2 ELISA method was 72.8%. In addition, 565 serum samples of cattle collected between 2017 and 2018 from four provinces in China were detected by the CLIA, and the seroprevalence for T. annulata ranged from 53.3% to 67.3% in these regions. Our findings demonstrated that the CLIA has high specificity, sensitivity and reliability, and could be used as a rapid detection assay for epidemiological investigations of T. annulata infection.


Acute acalculous cholecystitis in systemic lupus erythematosus: a rare initial manifestation.
Acute acalculous cholecystitis is a very rare gastrointestinal manifestation in systemic lupus erythematosus and becomes rarer as an initial manifestation. There are only two cases reported. The authors report a 20-year-old black woman that presented acute acalculous cholecystitis revealed by abdominal computed tomography. During hospitalization, she was diagnosed systemic lupus erythematosus. Conservative treatment with antibiotics was performed with complete remission of the symptoms. Corticosteroid was started in ambulatory. Cholecystectomy has been the treatment of choice in acute acalculous cholecystitis as a complication of systemic lupus erythematosus. The patient responded well to conservative treatment, and surgery was not required. This case is unique in the way that corticosteroid was started in ambulatory care. We should not forget that the acute acalculous cholecystitis can be the initial presentation of systemic lupus erythematosus although its occurrence is very rare. Conservative treatment should be considered. Abdominal computed tomography was a determinant exam for better assessment of acute acalculous cholecystitis.


Cold atmospheric nitrogen plasma induces metal-initiated cell death by cell membrane rupture and mitochondrial perturbation.
Cold atmospheric plasma (CAP) is a novel biomedical tool used for cancer therapy. A device using nitrogen gas (N2 CAP) produced CAP that induced cell death through the production of reactive nitrogen species and an increase in intracellular calcium. In this study, we investigated the effect of N2 CAP-irradiation on cell membrane and mitochondrial function in human embryonic kidney cell line 293T. We investigated whether iron is involved in N2 CAP-induced cell death, as deferoxamine methanesulfonate (an iron chelator) inhibits this process. We found that N2 CAP induced cell membrane disturbance and loss of mitochondrial membrane potential in an irradiation time-dependent manner. BAPTA-AM, a cell-permeable calcium chelator, inhibited N2 CAP-induced loss of mitochondrial membrane potential. These results suggest that disruption of intracellular metal homeostasis was involved in N2 CAP-induced cell membrane rupture and mitochondrial dysfunction. Moreover, N2 CAP irradiation generated a time-dependent production of peroxynitrite. However, lipid-derived radicals are unrelated to N2 CAP-induced cell death. Generally, N2 CAP-induced cell death is driven by the complex interaction between metal movement and reactive oxygen and nitrogen species produced by N2 CAP.


Nuclear import strategies of high risk HPV16 L1 major capsid protein.
During the late phase of human papillomavirus (HPV) infection, the L1 major capsid proteins enter the nuclei of host epithelial cells and, together with the L2 minor capsid proteins, assemble the replicated viral DNA into virions. We investigated the nuclear import of the L1 major capsid protein of high risk HPV16. When digitonin-permeabilized HeLa cells were incubated with HPV16 L1 capsomeres, the L1 protein was imported into the nucleus in a receptor-mediated manner. HPV16 L1 capsomeres formed complexes with Kap alpha2beta1 heterodimers via interaction with Kap alpha2. Accordingly, nuclear import of HPV16 L1 capsomeres was mediated by Kap alpha2beta1 heterodimers, required RanGDP and free GTP, and was independent of GTP hydrolysis. Remarkably, HPV16 L1 capsomeres also interacted with Kap beta2 and binding of RanGTP to Kap beta2 did not dissociate the HPV16 L1.Kap beta2 complex. Significantly, HPV16 L1 capsomeres inhibited the nuclear import of Kap beta2 and of a Kap beta2-specific M9-containing cargo. These data suggest that, during the productive stage of infection, while the HPV16 L1 major capsid protein enters the nucleus via the Kap alpha2beta1-mediated pathway to assemble the virions, it also inhibits the Kap beta2-mediated nuclear import of host hnRNP A1 protein and, in this way, favors virion formation.


[Role of Cahuzac's operation in clubfoot varus equina revision surgery].
The authors report 24 cases of revision in recurrent club foot deformity. They assessed Cahuzac's procedure for treatment of the residual forefoot adduction. This procedure (opening of the first cuneo-metatarsal joint and proximal abduction osteotomy of the second, third, and fourth metatarsals) was generally associated with postero-medial, plantar release, and split tibialis anterior tendon transfer. Twenty four procedures for 20 children at mean age of five years have been performed. The revision chart comprised 12 clinical and radiological items as proposed by Seringe. Seven feet were considered as excellent, 14 as good, 2 as fair, and 1 as poor. The mean follow-up was four years. Treatment of the adduction component with Cahuzac procedure is focused on the fore part of the foot, and on the calcaneo-pedal block by postero medial release, but never on the midfoot, as no Evans or Lichtblau's procedure has been performed. This series was compared to others procedures, and morphological results were equivalent. Cahuzac's operation is riskless for foot growth, and needs two approaches which can be useful for simultaneous procedures as split lateral transfer of tibialis anterior tendon. Metatarsal osteotomies (Cahuzac's procedure), associated with soft tissues release and split anterior tibial transfer, seems to be effective in surgical treatment of relapsed clubfeet, but the debate concerning the location of the adduction component of the deformity remains still open.


Statistical Thermodynamics for Actin-Myosin Binding: The Crucial Importance of Hydration Effects.
Actomyosin is an important molecular motor, and the binding of actin and myosin is an essential research target in biophysics. Nevertheless, the physical factors driving or opposing the binding are still unclear. Here, we investigate the role of water in actin-myosin binding using the most reliable statistical-mechanical method currently available for assessing biomolecules immersed in water. This method is characterized as follows: water is treated not as a dielectric continuum but as an ensemble of molecules; the polyatomic structures of proteins are taken into consideration; and the binding free energy is decomposed into physically insightful entropic and energetic components by accounting for the hydration effect to its full extent. We find that the actin-myosin binding brings large gains of electrostatic and Lennard-Jones attractive interactions. However, these gains are accompanied by even larger losses of actin-water and myosin-water electrostatic and LJ attractive interactions. Although roughly half of the energy increase due to the losses is cancelled out by the energy decrease arising from structural reorganization of the water released upon binding, the remaining energy increase is still larger than the energy decrease brought by the gains mentioned above. Hence, the net change in system energy is positive, which opposes binding. Importantly, the binding is driven by a large gain of configurational entropy of water, which surpasses the positive change in system energy and the conformational entropy loss occurring for actin and myosin. The principal physical origin of the large water-entropy gain is as follows: the actin-myosin interface is closely packed with the achievement of high shape complementarity on the atomic level, leading to a large increase in the total volume available to the translational displacement of water molecules in the system and a resultant reduction of water crowding (i.e., entropic correlations among water molecules).


[Comparison of coronary risk estimates derived using the Framingham and REGICOR equations].
To compare two equations for evaluating coronary risk, the Framingham-Wilson equation and the Framingham equation adjusted for the Spanish population (REGICOR), in a group of dyslipidemic patients in our healthcare area. In addition, the therapeutic implications of using the 2 methods were also evaluated. The study included 815 dyslipidemic patients, aged 35-74 years, from our healthcare area. Coronary risk was determined using the 2 equations and subjects were categorized as either low-risk (0%-9%), moderate-risk (10%-19%), or high-risk (> or =20%). To compare the application of the 2 equations, we evaluated differences in derived scores, coronary risk category, and the number of patients regarded as potentially treatable with hypolipidemic drugs. The best correlation observed between the 2 methods was for quantitative scores (r=0.983; P<.001). The correlation was poorer when coronary risk categories were compared (r=0.489; P<.001). Overall, the concordance was poor (kappa=0.06), and was only acceptable for low-risk patients (kappa=0.53). The coronary risk estimates derived from the Wilson table were 2.4 times higher than those obtained using REGICOR. The main differences were for moderate and high-risk patients. In addition, the number of patients regarded as potentially treatable with hypolipidemic drugs was five times higher when the Wilson equation was used. The overestimate of coronary risk obtained using the Framingham-Wilson equation leads to a greater number of patients being regarded as candidates for hypolipidemic treatment. Our data show the importance of using tables adjusted for the Spanish population.


Elimination of negative feedback in TLR signalling allows rapid and hypersensitive detection of microbial contaminants.
The exquisite specificity of Toll-like receptors (TLRs) to sense microbial molecular signatures is used as a powerful tool to pinpoint microbial contaminants. Various cellular systems, from native human blood cells to transfected cell lines exploit TLRs as pyrogen detectors in biological preparations. However, slow cellular responses and limited sensitivity have hampered the replacement of animal-based tests such as the rabbit pyrogen test or lipopolysaccharide detection by Limulus amoebocyte lysate. Here, we report a novel human cell-based approach to boost detection of microbial contaminants by TLR-expressing cells. By genetic and pharmacologic elimination of negative control circuits, TLR-initiated cellular responses to bacterial molecular patterns were accelerated and significantly elevated. Combining depletion of protein phosphatase PP2ACA and pharmacological inhibition of PP1 in the optimized reporter cells further enhanced the sensitivity to allow detection of bacterial lipoprotein at 30 picogram/ml. Such next-generation cellular monitoring is poised to replace animal-based testing for microbial contaminants.


Operative salvage for retroperitoneal nodal recurrence in colorectal cancer: a systematic review.
Retroperitoneal nodal recurrence after curative resection of colorectal cancer is an uncommon and challenging problem. The evidence for salvage surgery is limited and remains controversial, particularly when major vascular structures are involved. Some reports have demonstrated a survival benefit after metachronous resection of retroperitoneal metastasis with and without concomitant aortic resection. We conducted a systematic review of the literature to find evidence in favor of or against salvage surgery. Electronic searches of the MEDLINE, Cochrane, and EMBASE database were performed. Additional papers were identified by a manual search of the references from the key articles. Only peer-reviewed articles published in the English language were evaluated. A total of nine suitable studies were identified: three case reports and six larger series, of which one was a case-control study. Including our case reports, the total number of patients who underwent surgical resection that are available for review was 110. Median overall survival was between 34 and 44 months and median disease-free survival between 17 and 21 months. Concomitant resection of major vessels with graft replacement was feasible with survival ranging from 19 months to 18 years. There was no reported mortality associated with surgical salvage of retroperitoneal recurrence and the overall morbidity was 17-33%. The current literature suggests that more aggressive surgical treatment of retroperitoneal nodal recurrence in CRC has acceptable morbidity and may be associated with an improved survival in well-selected patients.


Sex differences in response rates during random ratio acquisition: effects of gonadectomy.
Sex differences in response rates were observed during acquisition of behavior on different random ratio schedules of reinforcement. Intact male rats responded at higher rates than intact female rats. Response rates of castrated males were found to be lower than response rates of intact males. Differences were not observed between intact and ovariectomized females.


Characterization of vasopressin and oxytocin immunoreactivity in the sheep and rat pineal gland: absence of vasotocin and detection of a vasopressin-like peptide.
The presence of arginine-vasopressin (VP), arginine-vasotocin (VT) and oxytocin (OT) were studied in sheep and rat pineal gland by the combination of HPLC and radioimmunoassays. Three immunoreactive substances, which had the same retention time as OT, VP and VT, were detected in sheep and rat pineal extracts after HPLC separation. Parallelism of dilution curve and standard curve demonstrated that two of them were identical to OT and VP respectively. The third component was distinct from VT on basis of its immunological reaction with two different antibodies; it resembled the immunoreactivity of VP. This substance was also detected in the hypothalamus, brain cortex and pituitary gland, but not in the hippocampus and adrenal gland. The results support the notion that VT is absent in the adult mammalian pineal gland and point to the existence of another, possibly related peptide.


Prognostic Nomogram for Patients with Hepatocellular Carcinoma After Thermal Ablation.
To develop an effective prognostic nomogram for patients with hepatocellular carcinoma (HCC) after thermal ablation. A total of 772 patients with intrahepatic primary or recurrent HCC who underwent radiofrequency ablation or microwave ablation between March 2011 and October 2016 were included. 602 patients (mean age, 56.0 ± 11.9 years; 495 male/107 female) were included in the primary cohort to establish a prognostic nomogram. Significant prognostic factors for overall survival (OS) identified by Cox univariate and multivariate regression analyses were used to construct the nomogram. The remaining 170 patients (mean age, 55.9 ± 11.9 years; 145 male/25 female) were used to validate the predictive accuracy of the nomogram. During a mean follow-up period of 26 months (range 1-85 months), the median OS periods were 48.6 months and 44.0 months for the primary and validation cohorts. The 1-, 3-, and 5-year OS rates were 85.5%, 61.4%, and 43.3% in the primary cohort and 84.7%, 59.6%, and 43.3% in the prospective validation cohort, respectively. Multivariate analysis found that pre-ablation treatment, AFP, CEA, CA19-9, ALBI grade, tumor number, and tumor size (hazard ratio > 1, P < 0.05) were independent risk factors for OS. A nomogram was developed based on these seven variables. The calibration curve for predicting the probability of survival showed a good agreement between the nomogram and actual observation both in the primary (concrete index: 0.699) and validation cohorts (concrete index: 0.734). This simple nomogram based on seven variables including ALBI grade offers personalized prognostic data for HCC patients after ablation. Level 4, case series.


SIRT3 deficiency exacerbates fatty liver by attenuating the HIF1α-LIPIN 1 pathway and increasing CD36 through Nrf2.
Deficiency of mitochondrial sirtuin 3 (SIRT3), a NAD+-dependent protein deacetylase that maintains redox status and lipid homeostasis, contributes to hepatic steatosis. In this study, we investigated additional mechanisms that might play a role in aggravating hepatic steatosis in Sirt3-deficient mice fed a high-fat diet (HFD). Studies were conducted in wild-type (WT) and Sirt3-/- mice fed a standard diet or a HFD and in SIRT3-knockdown human Huh-7 hepatoma cells. Sirt3-/- mice fed a HFD presented exacerbated hepatic steatosis that was accompanied by decreased expression and DNA-binding activity of peroxisome proliferator-activated receptor (PPAR) α and of several of its target genes involved in fatty acid oxidation, compared to WT mice fed the HFD. Interestingly, Sirt3 deficiency in liver and its knockdown in Huh-7 cells resulted in upregulation of the nuclear levels of LIPIN1, a PPARα co-activator, and of the protein that controls its levels and localization, hypoxia-inducible factor 1α (HIF-1α). These changes were prevented by lipid exposure through a mechanism that might involve a decrease in succinate levels. Finally, Sirt3-/- mice fed the HFD showed increased levels of some proteins involved in lipid uptake, such as CD36 and the VLDL receptor. The upregulation in CD36 was confirmed in Huh-7 cells treated with a SIRT3 inhibitor or transfected with SIRT3 siRNA and incubated with palmitate, an effect that was prevented by the Nrf2 inhibitor ML385. These findings demonstrate new mechanisms by which Sirt3 deficiency contributes to hepatic steatosis. Video abstract.


IGF-1 signaling mediated cell-specific skeletal mechano-transduction.
Mechanical loading preserves bone mass and stimulates bone formation, whereas skeletal unloading leads to bone loss. In addition to osteocytes, which are considered the primary sensor of mechanical load, osteoblasts, and bone specific mesenchymal stem cells also are involved. The skeletal response to mechanical signals is a complex process regulated by multiple signaling pathways including that of insulin-like growth factor-1 (IGF-1). Conditional osteocyte deletion of IGF-1 ablates the osteogenic response to mechanical loading. Similarly, osteocyte IGF-1 receptor (IGF-1R) expression is necessary for reloading-induced periosteal bone formation. Transgenic overexpression of IGF-1 in osteoblasts results in enhanced responsiveness to in vivo mechanical loading in mice, a response which is eliminated by osteoblastic conditional disruption of IGF-1 in vivo. Bone marrow derived stem cells (BMSC) from unloaded bone fail to respond to IGF-1 in vitro. IGF-1R is required for the transduction of a mechanical stimulus to downstream effectors, transduction which is lost when the IGF-1R is deleted. Although the molecular mechanisms are not yet fully elucidated, the IGF signaling pathway and its interactions with potentially interlinked signaling cascades involving integrins, the estrogen receptor, and wnt/β-catenin play an important role in regulating adaptive response of cancer bone cells to mechanical stimuli. In this review, we discuss recent advances investigating how IGF-1 and other interlinked molecules and signaling pathways regulate skeletal mechano-transduction involving different bone cells, providing an overview of the IGF-1 signaling mediated cell-specific response to mechanical stimuli. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:576-583, 2018.


Sources of error in the measurement of aortic diameter in computed tomography scans.
This study was conducted to determine the differences in the diameter of the thoracic aorta when measured from electrocardiographic (ECG)-gated and nongated computed tomography (CT) angiography. Another aim was to define the difference in the aortic diameter when it is measured at peak systole and end diastole in ECG-gated scans. The gated and nongated CT angiograms of 27 patients (mean age, 58 ± 16 standard deviation [SD] years) obtained on a 256-slice multidetector CT scanner were used. The transverse and anteroposterior diameters and the lumen areas were measured at 1, 4, and 8 cm below the origin of the left subclavian artery. There was a significant difference in the aortic measurements of diameter between gated and nongated scans found in samples taken at 1, 4, and 8 cm distal to the left subclavian artery (P < .0001). We found a considerable difference between the systolic and diastolic diameters (P < .0001). The maximum change in diameter between systole and diastole was 2.9 ± 0.9 (SD) mm (14.5%, P < .0001) at 1 cm, 5.4 mm (22.6%; median, 1.7 mm; P < .0001) at 4 cm, and 4.4 mm (16.9%; median, 1.3 mm; P < .0001) at 8 cm. There was a significant difference between the transverse and anteroposterior diameters in systole and diastole at all locations (P < .0001): The maximum change in diameter between transverse and anteroposterior diameters in systole was 5.4 ± 1.1 (SD) mm (15.7%, P < .0001) at 1 cm, 5.8 mm (19%; median, 1.4 mm; P < .0001) at 4 cm, and 5 mm (15%; median, 1.02 mm; P < .0001) at 8 cm. There was also a substantial difference between measuring the transverse diameter directly and deriving it from the lumen area (P < .0001). Our results showed an important difference between systolic and diastolic diameters measurements in ECG-gated scans. The standard protocol for measuring aortic diameters in gated scans of the thoracic aorta uses images at end diastole because the lack of wall motion at this time provides better resolution. This is likely to result in undersizing that, in some instances, may threaten stability and the proper seal of the stent graft. The dimensions of the aorta in a gated CT should be measured at peak systole rather than the conventional end diastole used today. Most medical centers use nongated CT or gated CT scans in end diastole to calculate sizes of endografts. In view of our findings, the latter method could result in potential complications.


May-Hegglin anomaly in pregnancy. A case report.
The May-Hegglin anomaly is a rare autosomal dominant platelet disorder that can be associated with significant bleeding. We treated a patient for pregnancy complicated by the May-Hegglin anomaly and a complete placenta previa.


The management of ankle fractures in the elderly.
In recent years, the incidence and severity of ankle fractures in the elderly population have increased. Although surgical fixation has gained wide acceptance for younger ankle fracture patients, controversy exists within the orthopaedic community with respect to the optimal way to manage these fractures in the geriatric patient population. Although some authors categorise ankle fractures in the elderly as fragility fractures associated with osteoporosis, it appears that risk factors such as increased weight, poly-pharmacy and propensity for falls play larger roles than poor bone quality. The presence of osteoporosis may increase the level of difficulty involved with the surgical management of these patients, leading some authors to alter their standard operative technique. Early studies cited high complication rates and poor outcome following operative intervention, however, more recent investigations have demonstrated successful functional outcomes following surgical management and appropriate postoperative rehabilitation. Based on the current evidence, the literature appears to support surgical fixation of displaced ankle fractures in the elderly patient population.


[Urinary tract injuries in polytraumatized patients].
Within the S3 Guideline Project of the European Association of Urology (EAU) an expert committee was set up to develop guidelines for the appropriate management of genitourinary trauma. These European guidelines were accepted in principle as national guidelines by the German Urological Society. Therefore, they also became the basis of the contribution of the German Urological Society to the S3 Guideline Project "Polytrauma" of the German Society for Trauma Surgery. For the guideline "management of genitourinary trauma" all the requirements for classification as S3 guidelines were full-filled. The guideline itself was developed in accordance with the principles of "evidence-based medicine". A systematic analysis of literature published between 1966 and 2004 was carried out. The articles retrieved were assessed in respect of study design and clinical relevance and classified following the scheme of the Centre for Evidence-Based Medicine in Oxford. In suspected renal injuries the hemodynamic situation of the patient is the benchmark for the diagnostic and therapeutic algorithm. The diagnostic gold standard for the assessment of haemodynamically stable patients is CT scanning. Uncontrolled haemodynamic instability is an indication for immediate explorative laparotomy. Partial ureteral tears are managed by stenting; complete tears by immediate surgical repair. Pelvic fractures are often associated with bladder ruptures. Extraperitoneal bladder ruptures, identified by retrograde cystography, are in most cases safely managed by simple catheter drainage. Intraperitoneal ruptures require surgical intervention. Blood at the meatus may suggest a urethral lesion-blind urethral catheterization should not be attempted. Suprapubic cystostomy and delayed urethroplasty are recommended.


Is first pregnancy age associated with hypertension in the Chinese rural women population?
The purpose of this study was to investigate the relationship between first pregnancy age and hypertension later in the life of women from Chinese rural areas. In total, 13,493 women were enrolled in the Henan Rural Cohort study. Logistic regression and linear regression were used to evaluate the association between first pregnancy age and hypertension and blood pressure indicators [including systolic blood pressure (SBP), diastolic blood pressure (DBP), and mean arterial pressure (MAP)]. The restricted cubic spline was used to examine the dose-response relationship between the first pregnancy age and hypertension or blood pressure indicators. After adjusting for potential confounders, each 1-year increase in first pregnancy age was associated with a 0.221 mmHg increase in SBP values, a 0.153 mmHg increase in DBP values, and a 0.176 mmHg decrease in MAP values (all P < 0.05). The β of SBP, DBP, and MAP showed a trend of first increasing and then decreasing with increasing first pregnancy age and there was no statistical significance after first pregnancy age beyond 33 years on SBP, DBP, and MAP, respectively. A 1-year increment in first pregnancy age was associated with a 2.9% [OR (95% CI): 1.029 (1.010, 1.048)] higher odds of prevalent hypertension. The odds of hypertension increased sharply and then eventually leveled off with an increment of first pregnancy age after adjusting for potential confounders. First pregnancy age might increase the risk of hypertension later in life and might be an independent risk factor for hypertension in women.


[Relationships between the value of combined PAH-inulin clearance, the kidney-cortex index as well as between additional radiographic findings in chronic pyelonephritis].
In 85 patients with ascertained chronic pyelonephritis the relations between the measured values of the combined PAH-inulin-clearance, the index of the adrenal cortex and other radiological findings were investigated. Here the determination of the cortex of the adrenal cortex resulted in a better correlation with the sizes clarification values than the calculation of the size of the kidney. By variance analysis (F-test) significant differences between the individual function groups became clear.


[E. Merck and cocaine. On Sigmund Freud's cocaine studies and their relation to the Darmstadt industry].
Documents from the archives of the pharmaceutical company, E. Merck, Darmstadt, shed light on research, production, and marketing of cocaine and other coca alkaloids. When cocaine proved to be a local anaesthetic in 1884 the market expanded enormously. The production of E. Merck is compared with that of other companies in Germany and abroad. Freud, who published on cocaine from 1884 to 1887, was in contact with E. Merck and performed clinical studies for them as well as for an American company.


Use of proton pump inhibitors is associated with an increase in adverse cardiovascular events in patients with hemodialysis: Insight from the kids registry.
Proton pump inhibitors (PPIs) are known to increase the risk of mortality and cardiovascular events in the general population. However, in patients with maintenance hemodialysis, PPI effects are under investigated. We analyzed the risk of PPIs for cardiovascular events using the Kagoshima Dialysis (KIDS) registry, a prospective, multicenter, observational study in patients with maintenance hemodialysis in Japan. In all, 531 patients were enrolled from June 2015 to December 2018. One-year follow-up data were available for 376 patients (Use of PPIs at baseline (PPI group): 217 patients and without PPIs (No PPI group): 159 patients). The incidence of a composite outcome (all-cause mortality, non-fatal myocardial infarction, or non-fatal stroke) was higher in patients in the PPI group than the No PPI group (15.2% vs. 4.4%; hazard ratio (HR): 3.65, 95% confidence interval (CI): 1.61-8.23, P = 0.002). In the multivariate analysis, even after adjustment for covariates, the use of PPIs was an independent risk factor for a composite outcome (HR: 2.38, 95% CI: 1.02-5.54, P = 0.045). We performed propensity score matching analysis as a sensitivity analysis, showing a consistent result. The incidence of bleeding showed no difference between the two groups (15.7% vs. 11.3%; HR: 1.46, 95% CI: 0.83-2.59, P = 0.19). These results indicate that the use of PPIs in patients with maintenance hemodialysis might increase mortality and cardiovascular events without decreasing the risk of bleeding. Therefore, it should always be analyzed if a patient truly needs PPIs.


Development of TaqMan Probe-Based One-Step RT-qPCR Assay Targeting 2B-NSP Coding Region for Diagnosis of Foot-and-Mouth Disease in India.
A one-step TaqMan probe-based RT-qPCR assay in the duplex format simultaneously targeting FMD Virus (FMDV) 2B NSP-coding region and 18S rRNA housekeeping gene was developed and evaluated. The duplex RT-qPCR assay specifically detected FMDV genome in both infected cell culture suspensions and a variety of clinical samples such as FMD-affected tongue/feet epithelium, oral/nasal swabs, milk and oro-pharyngeal fluids. The RT-qPCR assay was found to be highly sensitive, since the assay was 105-fold more sensitive than the traditional FMDV detecting antigen-ELISA (Ag-ELISA) and 102-fold better sensitive than both virus isolation and agarose gel-based RT-multiplex PCR. In addition, the assay could detect up to 100 copies of FMDV genome per reaction. In the epithelial samples (n = 582) collected from the FMD-affected animals, the diagnostic sensitivity was 100% (95% CI 99-100%). Similarly, all the FMDV-negative samples (n = 65) tested were confirmed negative by the new RT-qPCR assay, corresponding to 100% diagnostic specificity (95% CI = 94-100%). Further, the duplex RT-qPCR assay proved to be robust, showing an inter-assay co-efficient of variations ranging from 1.4 to 3.56% for FMDV-2B gene target, and from 2 to 4.12% for 18S rRNA gene target. While analyzing FMDV-infected cell culture suspension, a fairly strong positive correlation (correlation coefficient = 0.85) was observed between 2B-based RT-qPCR and WOAH-approved 5'UTR RT-qPCR assays. Therefore, the one-step RT-qPCR assay developed here with an internal control could be used for rapid, effective, and reliable detection of FMDV in pan-serotypic manner, and has the potential for routine diagnosis of FMDV in high throughput manner.


Effects of transport stress on the oxidative index, apoptosis and autophagy in the small intestine of caprine.
Introducing new goat breeds or transferring adult goats from farms to slaughterhouses requires transportation, which can engender adverse effects, such as oxidative stress, pathological cell apoptosis and autophagy. Current evidence suggests that malondialdehyde (MDA) is a metabolite of lipid peroxidation during oxidative stress, while superoxide dismutase (SOD) and catalase (CAT) can alleviate injury caused by free radicals and reactive oxygen species (ROS). Meanwhile, Bcl-2, Bax, LC3B, PINK1 and Parkin are important proteins that participate in pathological cell apoptosis and autophagy. This study aimed to investigate the effects of transportation stress on oxidative stress indexes and expressions of Bcl-2, Bax, LC3B, PINK1 and Parkin in the small intestine of goats. Twelve healthy adult male goats from western Jiangxi province were randomly divided into control, 2 h transportation stress, and 6 h transportation stress groups (n = 4 per group). Our results showed that MDA in the small intestine significantly increased after transportation, while SOD and CAT activities decreased, with a significantly increased apoptosis rate of the small intestine cells. The jejunum and duodenum exhibited the highest apoptosis rate in the 2 h and 6 h transportation groups, respectively. The expression of apoptosis-related genes Bcl-2 and Bax and their corresponding proteins exhibited varying degrees of down-regulation or up-regulation, while Bcl-2 and Bax genes in the small intestine were upregulated in the 6 h transportation group. In addition, autophagosomes and autophagolysosomes were found in various parts of the small intestine by transmission electron microscopy, and autophagy-related genes LC3B, PINK1 and Parkin were significantly down-regulated in the 2 h group and up-regulated in the 6 h group. Our results indicate that the contents of MDA, SOD and CAT in the small intestine, the expression of pathologic apoptosis-related genes Bcl-2 and Bax, and autophagy-related genes LC3B, PINK1 and Parkin correlated with stress duration caused by transportation. Moreover, this study provides a foothold for further studies on the mechanism of transportation stress in goats and improving animal welfare.


Characterising legacy spent nuclear fuel pond materials using microfocus X-ray absorption spectroscopy.
Analysis of a radioactive, coated concrete core from the decommissioned, spent nuclear fuel cooling pond at the Hunterston-A nuclear site (UK) has provided a unique opportunity to study radionuclides within a real-world system. The core, obtained from a dividing wall and sampled at the fill level of the pond, exhibited radioactivity (dominantly (137)Cs and (90)Sr) heterogeneously distributed across both painted faces. Chemical analysis of the core was undertaken using microfocus spectroscopy at Diamond Light Source, UK. Mapping of Sr across the surface coatings using microfocus X-ray fluorescence (μXRF) combined with X-ray absorption spectroscopy showed that Sr was bound to TiO2 particles in the paint layers, suggesting an association between TiO2 and radiostrontium. Stable Sr and Cs sorption experiments using concrete coupons were also undertaken to assess their interactions with the bulk concrete in case of a breach in the coating layers. μXRF and scanning electron microscopy showed that Sr was immobilized by the cement phases, whilst at the elevated experimental concentrations, Cs was associated with clay minerals in the aggregates. This study provides a crucial insight into poorly understood infrastructural contamination in complex systems and is directly applicable to the UK's nuclear decommissioning efforts.


WF-3405, a novel antitumor antibiotic. Taxonomy, isolation, structure elucidation and biological properties.
A new antitumor antibiotic, WF-3405 was isolated from the culture of Amauroascus aureus F-3405. The structure has been determined as 1,5-dioxiranyl-1,2,3,4,5-pentanepentanol on the basis of spectroscopic and chemical evidence. WF-3405 exhibits strong inhibitory activity against various murine tumors including leukemia P388, leukemia L1210 and Lewis lung carcinoma.


Specific proteins identified in whole saliva from patients with diffuse systemic sclerosis.
To evaluate the global changes of salivary protein profiles in patients with systemic sclerosis (SSc) using a proteomic approach. Whole saliva (WS) was collected from 15 patients with diffuse SSc and 15 healthy volunteers. Protein expression profiles for each sample were generated by 2-dimensional gel electrophoresis, and protein spots of interest were identified using peptide mass fingerprinting. The level of all the most representative salivary proteins except keratin 6L remained unchanged and only qualitative differences were observed between control subjects and patients with SSc. A total of 19 spots were found in SSc that were not matched with the controls. Fourteen out of a total of 19 spots were identified by mass analysis and were found to collapse into 9 unique proteins. These spots were identified to be cyclophilin A, calgranulin B, psoriasin, beta2-microglobulin, calgranulin A, glyceraldehyde-3-phosphate dehydrogenase, triose phosphate isomerase (TPI), actin-related protein 2/3 complex subunit 2 (Arp2/3 complex), and cystatin B. Our study is the first reporting the WS protein pattern of patients with SSc and comparing the differences between WS of patients with SSc and WS of healthy subjects. Both previously identified and newly identified proteins were detected in WS using a proteomic approach. Some of these proteins, like keratin 6L, psoriasin, TPI, and Arp2/3 complex, might have a pathological significance for SSc. It is possible that some of them can be defined as new therapeutic targets or diagnostic markers for SSc disease.


A study of the interaction between enantiomers of zolmitriptan and hydroxypropyl-beta-cyclodextrin by capillary electrophoresis.
The enantioresolution of zolmitriptan was performed using cyclodextrin (CD)-modified capillary zone electrophoresis (CZE) with hydroxypropyl-beta-CD (HP-beta-CD) as the chiral selector. The influence of experimental conditions on the enantioseparation of zolmitriptan, such as pH, temperature, applied voltage, and concentrations of running electrolyte and CD, was systematically investigated, obtaining a baseline separation of two enantiomers by the use of a 25 mM sodium dihydrogen phosphate (SDPH) running electrolyte (pH 2.4) containing 30 mM HP-beta-CD at 15 degrees C. Binding constants for each enantiomer-HP-beta-CD pair at different temperatures, as well as thermodynamic parameters for binding, were calculated. A nonlinear van't Hoff plot was obtained, indicating that the thermodynamic parameters of complexation were temperature-dependent for zolmitriptan enantiomers. The significant contribution of the enthalpy difference to the Gibbs free energy change suggested a stereomeric barrier mechanism for chiral recognition.


Antifibrotic effects of a herbal combination regimen on hepatic fibrotic rats.
Liver fibrosis has been characterized as chronic inflammatory processes involving multiple molecular pathogenetic pathways. This therapeutic study investigated whether a combination regimen of Salvia miltiorrhiza (S), Ligusticum chuanxiong (L) and Glycyrrhiza glabra (G) exerted in vivo antifibrotic effects on rats with hepatic fibrosis. Fibrosis was induced in rats by dimethylnitrosamine (DMN) administration for 4 weeks. Fibrotic rats were randomly assigned to one of the three groups: control, SLG (50 mg/kg) or silymarin (50 mg/kg), each given by gavage twice daily for 3 weeks starting 1 week after DMN injection. The results showed that fibrosis scores of livers from DMN-treated rats with SLG (1.13 +/- 0.13) were significantly reduced in comparison with DMN-treated rats receiving vehicle (1.63 +/- 0.18). Moreover, the hepatic collagen content of DMN rats was significantly reduced by either SLG or silymarin treatment. The double immunohistochemical staining results also showed that alpha-SMA positive cells with NF kappa B nuclear translocation were decreased in the fibrotic livers by SLG and silymarin treatments. The mRNA expression levels of TGF-beta1, alpha-SMA, collagen1 alpha 2, iNOS and ICAM-1 genes were attenuated by SLG and silymarin treatment. The results showed that SLG exerted antifibrotic effects in rats with DMN-induced hepatic fibrosis.


Naringenin and naringin in cardiovascular disease prevention: A preclinical review.
Cardiovascular disease is an important cause for morbidity and mortality worldwide. Flavonoids, such as naringin, and naringenin are important natural phytochemicals in the treatment or prevention of various disorders such as obesity, cardiac diseases, diabetes, and metabolic syndrome. Naringin and naringenin have significant therapeutic potential in several diseases through anti-oxidative, anti-inflammatory, and anti-apoptotic actions; these flavonoids play a protective role in human pathophysiology. In this review, based on the latest evidence, we present a summary of the impact of naringin, and naringenin on cardiovascular disease, and analyze and discuss the basic roles of naringin and naringenin and their mechanisms of actions in cardiovascular disease and other vascular dysfunction. The data collected in this review may serve as a comprehensive reference for the effects of naringin, and naringenin in cardiovascular disease, which may be beneficial for further research and for the design of naringin and naringenin analogs as new therapeutic options for cardiovascular diseases.


Projections of visceral and somatic primary afferents to the sacral spinal cord of the domestic fowl revealed by transganglionic transport of horseradish peroxidase.
The termination of visceral and somatic primary afferent fibers in the sacral spinal cord of the domestic fowl was studied using transganglionic transport of horseradish peroxidase. Nerve terminals of visceral afferent fibers were found in the lateral edge and base of the dorsal horn and in the dorsal gray commissure in close proximity to parasympathetic preganglionic neurons. Somatic afferents terminated in laminae 2 and 3. The results demonstrate that visceral and somatic afferent fibers in this avian species terminate in different areas of the dorsal horn.


Potent inhibition of drug-resistant HIV protease variants by monoclonal antibodies.
The monoclonal antibodies 1696 and F11.2.32 strongly inhibit the activity of wild-type HIV-1 protease (PR) by binding to epitopes at the enzyme N-terminus (residues 1-6) and flap residues 36-46, respectively. Here we demonstrate that these antibodies are also potent inhibitors of PR variants resistant to active-site inhibitors used as anti-AIDS drugs. Our in vitro experiments revealed that the inhibitory potency of single-chain fragments (scFv) of these antibodies is not significantly affected by the presence of mutations in PR; inhibition constants for drug-resistant protease variants are 5-11 nM and 13-169 nM for scFv1696 and for scFvF11.2.32, respectively. Tethered dimer of HIV-1 PR variant proved to be a model protease variant resistant to dissociative inhibition by 1696, and, strikingly, it also displayed resistance to inhibition by F11.2.32 suggesting that dimer dissociation also plays a role in the inhibitory action of F11.2.32.


cDNA cloning, expression, and mutagenesis study of liver-type prostaglandin F synthase.
Prostaglandin (PG) F synthase catalyzes the reduction of PGD2 to 9alpha,11beta-PGF2 and that of PGH2 to PGF2alpha on the same molecule. PGF synthase has at least two isoforms, the lung-type enzyme (Km value of 120 microM for PGD2 (Watanabe, K., Yoshida, R., Shimizu, T., and Hayaishi, O. (1985) J. Biol. Chem. 260, 7035-7041) and the liver-type one (Km value of 10 microM for PGD2 (Chen, L. -Y., Watanabe, K., and Hayaishi, O. (1992) Arch. Biochem. Biophys. 296, 17-26)). The liver-type enzyme was presently found to consist of a 969-base pair open reading frame coding for a 323-amino acid polypeptide with a Mr of 36,742. Sequence analysis indicated that the bovine liver PGF synthase had 87, 79, 77, and 76% identity with the bovine lung PGF synthase and human liver dihydrodiol dehydrogenase (DD) isozymes DD1, DD2, and DD4, respectively. Moreover, the amino acid sequence of the liver-type PGF synthase was identical with that of bovine liver DD3. The liver-type PGF synthase was expressed in COS-7 cells, and its recombinant enzyme had almost the same properties as the native enzyme. Furthermore, to investigate the nature of catalysis and/or substrate binding of PGF synthase, we constructed and characterized various mutant enzymes as follows: R27E, R91Q, H170C, R223L, K225S, S301R, and N306Y. Although the reductase activities toward PGH2 and phenanthrenequinone (PQ) of almost all mutants were not inactivated, the Km values of R27E, R91Q, H170C, R223L, and N306Y for PGD2 were increased from 15 to 110, 145, 75, 180, and 100 microM, respectively, indicating that Arg27, Arg91, His170, Arg223, and Asn306 are essential to give a low Km value for PGD2 of the liver-type PGF synthase and that these amino acid residues serve in the binding of PGD2. Moreover, the R223L mutant among these seven mutants especially has a profound effect on kcat for PGD2 reduction. The Km values of R223L, K225S, and S301R for PQ were about 2-10-fold lower than the wild-type value, indicating that the amino acid residues at 223, 225 and 301 serve in the binding of PQ to the enzyme. On the other hand, the Km value of H170C for PGH2 was 8-fold lower than that of the wild type, indicating that the amino acid residue at 170 is related to the binding of PGH2 to the enzyme and that Cys170 confer high affinity for PGH2. Additionally, the 5-fold increase in kcat/Km value of the N306Y mutant for PGH2 compared with the wild-type value suggests that the amino acid at 306 plays an important role in catalytic efficiency for PGH2.


Do Mitochondria Play a Central Role in Stress-Induced Somatic Embryogenesis?
This review highlights a four-step rational for the hypothesis that mitochondria play an upstream central role for stress-induced somatic embryogenesis (SE): (1) Initiation of SE is linked to programmed cell death (PCD) (2) Mitochondria are crucially connected to cell death (3) SE is challenged by stress per se (4) Mitochondria are centrally linked to plant stress response and its management. Additionally the review provides a rough perspective for the use of mitochondrial-derived functional marker (FM) candidates to improve SE efficiency. It is proposed to apply SE systems as phenotyping tool for identifying superior genotypes with high general plasticity under severe plant stress conditions.


Genetic Modification of Mesenchymal Stem Cells for Neurological Disease Therapy: What Effects Does it Have on Phenotype/Cell Behavior, Determining Their Effectiveness?
Mesenchymal stem cells are a promising tool in regenerative medicine, and their functions can be enhanced through genetic modification. Recent advances in genetic engineering provide several methods that enable gene delivery to mesenchymal stem cells. However, it remains to be decided whether genetic modification of mesenchymal stem cells by vectors carrying reporter or therapeutic genes leads to adverse effects on morphology, phenotypic profiles, and viability of transplanted cells. In this regard, we focus on the description of genetic modification methods of mesenchymal stem cells, their effectiveness, and the impact on phenotype/cell behavior/proliferation and the differentiation ability of these cells in vitro and in vivo. Furthermore, we compare the main effects of genetically modified mesenchymal stem cells with native mesenchymal stem cells when applied in the therapy of neurological diseases.


Influence of systemic factors on experimental epileptic brain injury. Structural changes accompanying bicuculline-induced seizures in rats following manipulations of tissue oxygenation or alpha-tocopherol levels.
A previous study from the laboratory showed that status epilepticus induced by bicuculline administration to ventilated rats produced astrocytic swelling and nerve cell changes ("type 1 and 2 injury") particularly in layers 3 and 5 of the neocortex (Söderfeldt et al. 1981). The type 1 injured neurons were characterized by condensation of cyto- and karyoplasm and the less common type 2 cells were characterized by swelling of endoplasmic reticulum including the nuclear envelope. In the present study we explored whether changes in cerebral oxygen availability altered the extent or character of the cellular alterations. Animals with 2 h of status epilepticus were made either hyperoxic (administration of 100% O2), hypoxic (arterial pO2 50 mm Hg) or hypotensive (arterial blood pressure of either 70-75 or 50 mm Hg). Furthermore, we explored whether "oxidative" damage occurred by manipulating tissue levels of alpha-tocopherol, a known free radical scavenger. Non-epileptic control animals exposed to comparable degrees of hypoxia or hypotension showed no or minimal structural alterations. In the epileptic animals the results were as follows. Hyperoxia did not change the quality or extent of the structural alterations previously observed in normoxic epileptic animals. Neither administration nor deficiency of vitamin E did modify this pattern of alterations. In hypoxia the extent of cell damage was the same or somewhat larger than in normoxic, epileptic animals. In addition, neurons often showed cytoplasmic microvacuoles due to swelling of mitochondria. The hypoxic animals also showed swelling of astrocytic nuclei with clumped chromatin. Changes similar to those observed in hypoxic animals also appeared in moderate hypotension (mean arterial blood pressure 50 mm Hg), whereas mild hypotension (70-75 mm Hg) did not change the character of the tissue injury from that seen in hyperoxic or normoxic epileptic rats. The present results demonstrate that the neuronal cell damage that can be observed when the brain is fixed by perfusion after status epilepticus of 2 h duration is not exaggerated by hyperoxia or vitamin E deficiency nor is it ameliorated by a moderate restriction in cerebral oxygen supply or by vitamin E administration. If anything, hypoxia (or moderate hypotension) appears to increase the extent of damage and it clearly alters its ultrastructural characteristics. However, although the results fail to support the notion that epileptic cell damage is "oxidative", definite conclusions must await information on the cell damage that remains upon arrest of the epileptic activity.


Frequency of alpha-particles from 239PuO2 in lung cells.
NCRP-Report 49 deals with the effect of 239PuO2-particles in the lung. A basic aspect in the considerations is the frequency of cells traversed by one or by more alpha-particles. The present article contains the explicit derivation of the formulae underlying the data in the NCRP-report.