Lipoxygenases (LOXs) are lipid-peroxidizing enzymes that consist of a regulatory calcium- and membrane-binding PLAT (polycystin-1, lipoxygenase, α-toxin) domain and a catalytic domain. In a previous study, the crystal structure of an 11R-LOX revealed a conserved π-cation bridge connecting these two domains which could mediate the regulatory effect of the PLAT domain to the active site. Here we analyzed the role of residues Trp107 and Lys172 that constitute the π-cation bridge in 11R-LOX along with Arg106 and Asp173-a potential salt bridge, which could also contribute to the inter-domain communication. According to our kinetic assays and protein unfolding experiments conducted using differential scanning fluorimetry and circular dichroism spectroscopy, mutants with a disrupted link display diminished catalytic activity alongside reduced stability of the protein fold. The results demonstrate that both these bridges contribute to the two-domain interface, and are important for proper enzyme activation.

Whereas nitrosation of secondary amines produces nitrosamines, amino acids with primary amino groups and glycine ethyl ester were reported to react with nitrite to give unidentified agents that alkylated 4-(p-nitrobenzyl)pyridine to produce purple dyes and be direct mutagens in the Ames test. We report here that treatment of glycine ethyl ester at 37 degrees C with excess nitrite acidified with HCl, followed by ether extraction, gave 30-40% yields of a product identified as ethyl chloro(hydroximino)acetate [ClC(=NOH)COOEt, ECHA] and a 9% yield of ethyl chloroacetate. The ECHA was identical to that synthesized by a known method from ethyl acetoacetate, strongly alkylated nitrobenzylpyridine, and may have arisen by N-nitrosation of glycine ethyl ester to give ethyl diazoacetate, which was C-nitrosated and reacted with chloride to give ECHA. Nitrosation of ethyl diazoacetate also yielded ECHA. Ethyl nitroacetate was not an intermediate as its nitrosation did not produce ECHA. ECHA reacted with aniline to give ethyl (hydroxamino)(phenylimino)acetate [PhN=C(NHOH)CO2Et]. This product was different from ethyl [(phenylamino)carbonyl]carbamate [PhNHC(=O)NHCO2Et], which was synthesized by reacting ethyl isocyanatoformate (OCN.CO2Et) with aniline. ECHA reacted with guanosine to give a derivative, which may have been a guanine-C(=NOH)CO2Et derivative. ECHA showed moderate toxicity and weak but significant mutagenicity without activation in Salmonella typhimurium TA-100 (mean, 1.31 x control value for 12-18 microg/plats) and for V79 mammalian cells (1.5-1.7 x control value for 60-100 microM). In conclusion, gastric nitrosation of glycine derivatives such as peptides with a N-terminal glycine might produce ECHA analogues that alkylate bases of gastric mucosal DNA and thereby initiate gastric cancer.

Background: Square knots can be difficult to construct in deep body cavities. The reversing half-hitch alternating post (RHAP) surgical knot has noninferior tensile strength and performance characteristics in deep body cavities. We compared the enterotomy repairs of novice learners in simulated deep body cavities using RHAP versus square knots after proficiency-based training.

Methods: Undergraduate students were randomized to RHAP (n = 10) or square knot (n = 10) groups and trained to defined proficiency. They then performed hand-sewn enterotomy repairs of cadaveric porcine small bowels on flat surfaces and in simulated deep body cavities. We recorded time to knot-tying proficiency and to enterotomy repair, and burst pressures for the repair.

Results: Mean time-to-proficiency in knot tying was equivalent between the RHAP and square knot groups (23 [standard deviation (SD) 3] v. 21 [SD 2] min, p = 0.33). Mean time for enterotomy repair in deep cavities was shorter for the RHAP group (16 [SD 2] min v. 21 [SD 1] min, p = 0.02). Mean burst pressures for enterotomy repair were equivalent on flat surfaces (128 [SD 41] v. 101 [SD 36] mm Hg, p = 0.31), and were significantly higher for the RHAP group in simulated deep body cavities (32 [SD 13] v. 105 [SD 37] mm Hg, p = 0.05).

Conclusion: The RHAP knots appear to have superior performance versus square knots when tied in a deep body cavity by novice learners. Future work should focus on demonstrating the clinical relevance and broad utility of the RHAP knot in abdominal surgery. Both knot types should be taught to novice learners.

Contexte: L’exécution de noeuds plats peut être difficile dans les cavités corporelles profondes. Les noeuds de type demi-clé inversée alternée (RHAP, pour reversing halfhitch alternating post) ont une résistance à la traction et un rendement semblables à ceux des noeuds plats dans ces cavités. Nous avons comparé l’efficacité des noeuds plats et des noeuds de type RHAP réalisés par de nouveaux apprenants dans des cavités profondes simulées, après leur avoir enseigné les compétences nécessaires.

Méthodes: Les étudiants de premier cycle ont été aléatoirement répartis en 2 groupes, soit le groupe RHAP (n = 10) et le groupe noeud plat (n = 10), et ont reçu une formation pour développer des compétences prédéfinies. Ils ont ensuite suturé à la main un intestin grêle provenant d’un cadavre de porc, sur une surface plane et à l’intérieur d’une cavité profonde simulée. Nous avons mesuré le temps nécessaire à l’exécution du noeud et à la suture complète de l’incision, de même que la pression que pouvait subir cette suture sans se rompre.

Résultats: Le temps moyen d’exécution du noeud était semblable entre les groupes RHAP et noeud plat (23 min [écart type (E.T.) 3 min] c. 21 min [E.T. 2 min]; p = 0,33). Le temps moyen nécessaire à la suture de l’incision dans la cavité profonde était plus court dans le groupe RHAP (16 min [E.T. 2 min] c. 21 min [E.T. 1 min]; p = 0,02). La pression moyenne que pouvait subir la suture sans se rompre était comparable pour les sutures effectuées sur une surface plane (128 mm Hg [E.T. 41 mm Hg] c. 101 mm Hg [E.T. 36 mm Hg]; p = 0,31), mais était significativement plus élevée dans le groupe RHAP pour les sutures faites dans la cavité profonde (32 mm Hg [E.T. 13 mm Hg] c. 105 mm Hg [E.T. 37 mm Hg], p = 0,05).

Conclusion: Les noeuds de type RHAP semblent avoir un rendement supérieur à celui des noeuds plats lorsqu’ils sont réalisés dans une cavité profonde par de nouveaux apprenants. Des études ultérieures devraient se pencher sur la pertinence clinique et l’utilité générale de ces noeuds en chirurgie abdominale. Les 2 types de noeuds devraient être enseignés aux nouveaux apprenants.

Purpose: The Hippo pathway has broadened in cancer research in the past decade and revealed itself to be an important driver for tumorigenesis and metastatic spread. In this study, we investigated the clinical relevance of the Hippo pathway with regard to metastatic invasion, patients' outcome and histopathological features.

Methods: Protein expression of components of the Hippo pathway were analyzed by immunohistochemistry (IHC) using paraffin-embedded tissue from 103 patients who had been diagnosed with pancreatic ductal adenocarcinoma and had undergone surgery. Results were correlated with clinicopathological data, disease-free and overall survival.

Results: Immunohistochemistry studies in pancreatic tumour tissues revealed a significant upregulation of MST1, MST2, pLATS, pYAP and 14-3-3, representing the active Hippo pathway, in non-metastasized patients (p < 0.01). In turn, the pathway is more inactive in metastasized patients and relating liver metastases as LATS1, LATS2, YAP, transcriptional factors TEAD2 and TEAD3 were upregulated in these patients (p < 0.01). A higher pYAP expression was associated with a favorable OS and DFS.

Conclusion: The Hippo pathway is inactive in metastasized patients releasing the pro-metastatic and proliferative potential of the pathway. Furthermore, our study underlines the prognostic relevance of the Hippo pathway as a shift in the balance towards the inactive pathway predicts an unfavorable OS and DFS.

Keywords: Hippo pathway; Metastasis; PDAC; Pancreatic cancer; Prognosis.

In preparation for the clinical use of Plat-II Castable Ceramics (PCC-II), we tested its compressive strength, tensile strength, flexural strength and microhardness. The flexural strength was tested by the three-point bending test. The result showed that the compressive strength was 541.7 MPa; the tensile strength 42.5 MPa; the flexural strength 142.0 MPa; the modulus of elasticity 61.4 GPa; and the micro-hardness 499.6 kg/mm3. The strength anol micro-hardness of PCC-II materials were better than those of human enamel. So PCC-II is applicable to restorations in dentistry.

The aim of this study was to determine the effect of temporary inhibition of meiosis using the cyclin-dependent kinase inhibitor butyrolactone I (BLI) on gene expression in bovine oocytes and cumulus cells. Immature bovine cumulus-oocyte complexes (COCs) were assigned to groups: (i) Control COCs collected immediately after recovery from the ovary or (ii) after in vitro maturation (IVM) for 24 h, (iii) Inhibited COCs collected 24 h after incubation with 100 μm BLI or (iv) after meiotic inhibition for 24 h followed by IVM for a further 22 h. For mRNA relative abundance analysis, pools of 10 denuded oocytes and respective cumulus cells were collected. Transcripts related to cell cycle regulation and oocyte competence were evaluated in oocytes and cumulus cells by quantitative real-time PCR (qPCR). Most of the examined transcripts were downregulated (p < 0.05) after IVM in control and inhibited oocytes (19 of 35). Nine transcripts remained stable (p>> 0.05) after IVM in control oocytes; only INHBA did not show this pattern in inhibited oocytes. Seven genes were upregulated after IVM in control oocytes (p < 0.05), and only PLAT, RBP1 and INHBB were not upregulated in inhibited oocytes after IVM. In cumulus cells, six genes were upregulated (p < 0.05) after IVM and eight were downregulated (p < 0.05). Cells from inhibited oocytes showed the same pattern of expression regarding maturation profile, but were affected by the temporary meiosis inhibition of the oocyte when the same maturation stages were compared between inhibited and control groups. In conclusion, changes in transcript abundance in oocytes and cumulus cells during maturation in vitro were mostly mirrored after meiotic inhibition followed by maturation.

Purpose of review: Chronic rhinosinusitis with nasal polyps (CRSwNP) is a common and heterogeneous inflammatory disease. The underlying epigenetic mechanisms and treatment of CRSwNP are partially understood. Of the different epigenetic changes in CRSwNP, histone deacetylases (HDACs), methylation of DNA, and the levels of miRNA are widely studied. Here, we review the human studies of epigenetic mechanisms in CRSwNP.

Recent findings: The promoters of COL18A1, PTGES, PLAT, and TSLP genes are hypermethylated in CRSwNP compared with those of controls, while the promoters of PGDS, ALOX5AP, LTB4R, IL-8, and FZD5 genes are hypomethylated in CRSwNP. Promoter hypermethylation suppresses the gene expression, while promoter hypomethylation increases the gene expression. Studies have shown the elevation in the levels of HDAC2, HDAC4, and H3K4me3 in CRSwNP. In CRSwNP patients, there is also an upregulation of certain miRNAs including miR-125b, miR-155, miR-19a, miR-142-3p, and miR-21 and downregulation of miR-4492. Epigenetics takes part in the immunology of CRSwNP and may give rise to endotypes of CRSwNP. Both HDAC2 and the miRNA including miR-18a, miR-124a, and miR-142-3p may take function in the regulation of glucocorticoid resistance. HDAC inhibitors and KDM2B have shown effectiveness in decreasing nasal polyp, and DNA methyltransferase (DNMT) or HDAC inhibitors may have a potential efficacy for the treatment of CRSwNP. Recent advances in the epigenetics of CRSwNP have led to the identification of several potential therapeutic targets for this disease. The use of epigenetics may provide novel and effective biomarkers and therapies for the treatment of nasal polyp.

Keywords: Chronic rhinosinusitis with nasal polyps (CRSwNP); Epigenetics; Histone deacetylase (HDAC); Methylation; Nasal polyp; miRNA.

Lipoxygenase-1 (LOX-1) protein provides defense against pests and pathogens and its presence have been positively correlated with plant resistance against pathogens. Linoleate is a known substrate of lipoxygenase and it induces necrosis leading to the accumulation of isoflavonoid phytoalexins in plant leaves. Therefore, it is of interest to study the structural features of LOX-1 from Finger millet. However, the structure ofLOX-1 from Finger millet is not yet known. A homology model of LOX-1 from Finger millet is described. Domain architecture study suggested the presence of two domains namely PLAT (Phospho Lipid Acyl Transferase) and lipoxygenase. Molecular docking models of linoleate with lipoxygenase from finger millet, rice and sorghum are reported. The features of docked models showed that finger millet have higher pathogen resistance in comparison to other cereal crops. This data is useful for the molecular cloning of fulllength LOX-1 gene for validating its role in improving plant defense against pathogen infection and for various other biological processes.

Gaseous Hg can evaporate and enter the plants through the stomata of plat leaves, which will cause a serious threat to local food safety and human health. For the risk assessment, this study aimed to characterize atmospheric mercury (Hg) as well as its accumulation in 5 leafy vegetables (spinach, edible amaranth, rape, lettuce, allium tuberosum) from sewage-irrigated area of Tianjin City. Bio-monitoring sites were located in paddy (wastewater irrigation for 30 a), vegetables (wastewater irrigation for 15 a) and grass (control) fields. Results showed that after long-term wastewater irrigation, the mean values of mercury content in paddy and vegetation fields were significantly higher than the local background value and the national soil environment quality standard value for mercury in grade I, but were still lower than grade II. Soil mercury contents in the studied control grass field were between the local background value and the national soil environment quality standard grade I . Besides, the atmospheric environment of paddy and vegetation fields was subjected to serious mercury pollution. The mean values of mercury content in the atmosphere of paddy and vegetation fields were 71.3 ng x m(-3) and 39.2 ng x m(-3), respectively, which were markedly higher than the reference gaseous mercury value on the north sphere of the earth (1.5-2.0 ng x m(-3)). The mean value of ambient mercury in the control grass fields was 9.4 ng x m(-3). In addition, it was found that the mercury content in leafy vegetables had a good linear correlation with the ambient total gaseous mercury (the data was transformed into logarithms as the dataset did not show a normal distribution). The comparison among 5 vegetables showed that the accumulations of mercury in vegetables followed this order: spinach>> edible amaranth>> allium tuberosum>> rape>> lettuce. Median and mean values of mercury contents in spinach and edible amaranth were greater than the hygienic standard for the allowable limit of mercury in food. Spinach appeared to accumulate more mercury than the other four vegetables, in which the median and mean mercury content were both higher than 20 μg x kg(-1). The mercury concentrations in rape, lettuce and allium tuberosum were lower than the standard. Moreover, test results indicated that the Hg content in leafy vegetables was mainly the gaseous mercury through leaf adsorption but not the Hg particulates. This study clearly manifested that there should be a great concern on the pollution risk of both air-and soil borne mercury when cultivating leafy vegetables in long-term wastewater-irrigated area.

The goal of the current work is to study the molecular mechanisms underlay the action of 5- amino-exo-3-azatricyclo[5.2.1.0(2,6)]decan-4-one (P-11) with combined antiarrhythmic, nootropic, anti-inflammatory and anaesthetic activities. The aconitine-induced experimental rat model of cardiac arrhythmia has been used in our study. Aconitine was administered once intravenously in a dose 50 microg/kg whereas experimental animal group received P-11 in a dose 0.3 mg/kg (the compound was injected intravenously 2 min before acute aconitine treatment). Expression macroarray (Atlas Rat cDNA Expression Array, #7738-1; BD Biosciences) was used to identify the target genes for P-11 compound. Comparative analysis of changes in the status of expression of genes in the heart of rats induced by P-11 against the simulated in vivo arrhythmia identified 16 genes that reproducibly alter the level of expression.These genes encode the extracellular matrix proteins (glypican 1, Gpc1; tissue inhibitor of metalloproteinase 2, 3, Timp2, Timp 3); intracellular signaling molecules (rho GTPase activating protein 7, Dlc1; protein tyrosine phosphatase 4a1, Ptp4a1; phosphodiesterase 4D, PDE4D; PI3-kinase regulatory subunit alpha, PIK3R1; guanine nucleotide binding protein alpha 12, Gna12) and protein of intermediate junctions (junction plakoglobin, Jup), proteins involved in glycolysis (phosphofructokinase I, Pfk1) and hemostasis (tissue plasminogen activator, Plat), plasma membrane transporters (Solute carrier family 16, member 1, Slc16a1; ATPase, Na+/K+ transporting, Atp1a), and ets. (c-fos protooncogene, c-fos; telomerase protein component 1, tlp; Annexin 1, anxa 1). Thus, the data about the selective effect of P-11 on genes whose products are involved in the aritmogenesys mechanisms, allow us to consider this compound as a promising means of pathogenetically oriented pharmacotherapy of cardiac arrhythmias.

Basal stem rot (BSR) is a devastating disease to Malaysian oil palm. Current techniques employed for BSR disease detection on oil palm are laborious, time consuming, costly, and subjected to accuracy limitations. An ergosterol detection method was developed, whereby it correlated well with the degree of infection in oil palm. This current study was designed to study the relationship between Ganoderma biomass, ergosterol concentration, BSR disease progress and to validate the efficiency of microwave assisted extraction (MAE) method for extraction of ergosterol compound. In addition, testing on the sensitivity of thin layer chromatography (TLC) analysis for detection of ergosterol was also the aim of this study. The optimised procedure involved extracting a small amount of Ganoderma-infected oil palm root tissues suspended in low volumes of solvent followed by irradiation in a conventional microwave oven at 70°C and medium high power for 30 s, resulting in simultaneous extraction and saponification. Based on the results obtained, MAE method may be effective in extracting low to high yields of ergosterol from infected oil palm roots demonstrating disease scale 2, 3 and 4. Positive relationship was observed between ergosterol content and inoculation period starting day 3 in the inoculated oil palm seedlings and hour 6 in germinated seeds. TLC analysis demonstrated a good correlation with high performance liquid chromatography (HPLC) quantification. Therefore, a semi-quantitative TLC analysis may be applied for handling a large amount of samples during onset field survey.

Reput pangkal batang (BSR) adalah penyakit yang membawa kemusnahan kepada industri sawit di Malaysia. Teknik semasa yang digunakan bagi mengesan penyakit BSR pada sawit adalah sukar, memakan masa, mahal, serta mempamerkan kadar ketepatan yang terbatas. Oleh itu, kaedah pengesanan ergosterol patogen telah dibangunkan, di mana ia berkorelasi baik dengan tahap jangkitan pada sawit. Kajian ini direka bagi mengkaji hubungan di antara jisim Ganoderma, kepekatan ergosterol, perkembangan penyakit BSR dan untuk mengesahkan kecekapan kaedah pengekstrakan berbantukan ketuhar gelombang mikro (MAE). Di samping itu, ujian terhadap kepekaan analisis plat kromatografi lapisan nipis (TLC) bagi pengesanan ergosterol juga merupakan tujuan kajian ini. Prosedur yang telah dioptimumkan melibatkan pengekstrakan sejumlah kecil tisu akar sawit yang telah dijangkiti patogen Ganoderma yang diampai dalam pelarut dengan isipadu rendah. Ini diikuti oleh penyinaran dalam ketuhar gelombang mikro konvensional pada suhu 70°C dan kuasa sederhana tinggi selama 30 s, menghasilkan pengekstrakan dan saponifikasi secara serentak. Berdasarkan keputusan yang diperolehi, kaedah MAE mungkin berkesan dalam mengekstrak ergosterol daripada kuantiti tisu sawit yang sedikit tetapi mampu menunjukkan tahap penyakit sawit dari skala 2, 3 dan 4. Hubungan positif diperhatikan di antara kandungan ergosterol dan tempoh inokulasi bermula hari ke-3 dalam benih sawit yang telah diinokulasi dan 6 jam selepas inokulasi bagi benih yang telah bercambah. Analisis TLC telah menunjukkan korelasi yang baik dengan kuantiti kromatografi cecair prestasi tinggi (HPLC). Oleh itu, analisis separa kuantitatif TLC boleh digunakan untuk mengendalikan sejumlah besar sampel semasa tinjauan di lapangan.

Keywords: Basal stem rot; Ganoderma; HPLC; Microwave assisted extraction; Oil palm; Pengekstrakan berbantukan ketuhar gelombang mikro; Reput pangkal batang; Sawit; TLC; Thin layer chromatography (TLC).

UNASSIGNED

Azomonas agilis, a nitrogen-fixing bacterium, was isolated from rhizospheric soil in central Myanmar.

UNASSIGNED

The nitrogen-fixing activity of this bacterium was detected by plate screening method using glucose nitrogen free mineral medium and ammonium test-kit Cellulolytic activity was screened by plat assay and detected by Dinitrosalicyclic acid method (DNS).

UNASSIGNED

The isolated A. agilis grew in media containing 3-12% of NaCl, although the growth became poor when NaCl concentrations increased. Among various carbon sources, sucrose was the best source for ammonium accumulation of this bacterium, whereas arabinose was not the suitable carbon source. Although the nitrogen-fixing activity of A. agilis was highest after one week incubation, cellulase enzyme production was highest after 2-3 days of incubation. It was observed that cellulase enzyme activity of A. agilis for cellulose and sodium carboxymethyl cellulose (CMC) was almost the same. Three agricultural wastes were used to detect the cellulase enzyme activity of A. agilis, cellulase activity was better on filter paper as a substrate when compared to rice-straw and sawdust.

UNASSIGNED

So, the isolated A. agilis has high potential as an effective bacterial strain to use in sustainable agriculture and degradation of some agricultural residues.

Antibody-drug conjugates (ADCs) are a therapeutic modality that traditionally enable the targeted deliv-ery of highly potent cytotoxic agents to specific cells such as tumor cells. More recently, antibodies have been used to deliver molecules such as antibiotics, antigens, and adjuvants to bacteria or specific immune cell subsets. Site-directed mutagenesis of proteins permits more precise control over the site and stoichiometry of their conjugation, giving rise to homogenous chemically defined ADCs. Identification of favorable sites for conjugation in antibodies is essential as reaction efficiency and product stability are influenced by the tertiary structure of Immunoglobulin G (IgG). Current methods to evaluate potential conjugation sites are time-consuming and labor intensive, involving multi-step process-es for individually produced reactions. Here, we describe a highly efficient method for identification of conjugatable genetic variants by analyzing pooled ADC libraries using mass spectrometry. This approach provides a versatile plat-form to rapidly uncover new conjugation sites for site-specific ADCs.

Background: Use of plants and plant products in health care has shown exponential increase in past two decades.

Introduction: In-spite of the availability of well-established pharmacotherapy for epilepsy, a large no of population still explores alternative treatments due to refractory seizures, adverse effects of drugs, chronic treatment, inaccessibility of standard therapies in rural areas and the social stigma attached to the disease. Various studies on medicinal plants showed the protective effect of herbals in animal models of epilepsy.

Method: In the present review, a status analysis of the traditional use of various medicinal plants in epilepsy with a special focus on plats having anti-inflammatory potential is recorded.

Result and conclusion: The shortcomings of research on medicinal plants which needs to be explored further in order to tackle the growing need of safer and effective drugs for epilepsy are discussed. Overall, there is a huge scope of herbal drugs in CNS disorders especially epilepsy, either as an adjunct by reducing the dose and thus side effects of standard anti-epileptic drugs or as standalone agent . Although, there is still an urgent need of well planned randomized controlled clinical trials to validate their efficacy and safety.

Keywords: CNS activity; epilepsy; herb-drug interaction; herbal drugs.

Microspectrophotometer and transmitted electron microscope (TEM) were used to study the change of DNA synthesis and ultrastructure after fibronection(FN) acted on cultured periodontal ligament cells (PDLC).The results showed that the data of nuclus area,nuclus total absorbs and nuclus mean absorbs of PDLC in FN group was higher than control(P<0.01).Under the TEM we observed that in FN group there were more euchromatin in cell nuclues and more organ cells such as microfilaments,chondriosomes and endoplasmic reticulum in cytoplasm.It suggested that FN can accelerate periodontal regeneration by promoting DNA and protein synthesis of PDLC.

OBJECTIVE

To improve the MigR1-CD19-CAR (chimeric antigen receptor) that contains a single chain variable region (scFv) which targeted to CD19 through a retroviral vector transduction efficiency of T-lymphocytes.

METHODS

Insert the CD19-CAR fragment into the retroviral vector (MigR1) through recombinant DNA technology, after transfecting plat-A packaging cell lines, viral supernatant was collected to transduce K562 cell line and activated human T-lymphocytes. We used flow cytometry to determine the transduction efficiency and RT-PCR to confirm the transcription of CD19-CAR gene. The ability of the transduced T cells to produce IFN-γ and TNF-α in a CD19-specific manner was measured in an enzyme-linked immunosorbent (ELISA) assay.

RESULTS

(1)Using MigR1-CD19-CAR retroviral vector to produce the high titer retrovirus. (2)MigR1-CD19-CAR transduction efficiency of K562 cell line was significantly higher than human T-lymphocytes (P<0.01). (3)120 min centrifugation could significantly improve transduction efficiency of T-lymphocytes to (54.5±14.6)%. (4)Transduction efficiency could be improved by deciding transduce time according to T-lymphocytes proliferation fold in vitro individually, and the highest transduction efficiency in the study was 69.3%. The CD19-CAR gene sequence was transcripted specificly with high efficiency. (5) IFN-γ and TNF-α released by CD19-CAR transduced T-lymphocytes significantly increased to (13 230±1 543) pg/ml and (4 217±211) pg/ml when coculture with CD19-K562 cells.

CONCLUSIONS

We have successfully constructed a second generation CAR which targeted to CD19 through a retroviral vector called MigR1 (MigR1-CD19-CAR). Deciding transduce time according to T-lymphocytes proliferation fold in vitro individually and 120 min centrifugation could improve the CAR transduction efficiency of T-lymphocytes. RT-PCR confirmed that the CD19-CAR gene was specificly transcripted with high efficiency. IFN-γ and TNF-α released by CD19-CAR transduced T-lymphocytes significantly increased when activated by target cells.

The paper deals with some less known ethnomedicinal plats as well as animals which are use I the treatment of piles by the Bhoxa tribe of Uttar Pradesh.

One of the main concerns of dental research is the observation of the oral tissues and the materials applied to the dentition. The changes in composition and structure of the outer surfaces and the materials deposited on these surfaces are of special interest. In the literature, a variety of replica techniques for these purposes is described (Grundy in 1971 [12]; Saxton in 1973 [25]). The use of these techniques is limited because of artifacts in the samples, and a restricted resolution power resulting from useful magnifications in the order of 800x. An accurate and universal replica technique for the examination of specimens to be viewed under the SEM has been developed. The first impression is made by a light body silicone elastomer (President Coltene). The positive replica is made by electrodeposition of copper in an electro plating bath (Acru plat 5 electronic, Dr. Th. Wieland, D-7530 Pforzheim). The reliability and accuracy of this replica technique was verified by a scanning electron microscopic comparison of the replicas and the actual structures of etched enamel. To illustrate the applicability of the replica technique to structures with much lower hardness, also high resolution images of dental plaque were produced. The copper surface offers a perfect, original and proper electroconductive medium that withstands the bombardment of electrons and the relatively severe conditions in the scanning electron microscope. Reproducibility was accurate as judged by the duplication in position, size, and shape of the fine detail at magnifications of 7500x offering a resolution of 25 nm.

The presence of prairies in Alabama's Black Belt as reported by early explorers is further substantiated here by using the field notes and plats of the original land survey of Montgomery County (1845-46). One-third of the approximately 200,000 acres of Black Belt in the county was depicted as prairie on the field plats. A strong correlation was found between rates of low tree density (10 trees or less per acre) as given in the field notes and areas shown as prairies on the plants (one tree per acre equals 2.47 per hectare). Areas of low tree density and prairies, when located on recent soil maps (1957), occurred primarily on upland alkaline soils. Apparently these upland alkaline sites composed the true prairie and were those described as savannahs," plains native fields, and prairies by the early explorers. High tree densities (120 trees or more per acre) were associated primarily with areas outside prairies and were mostly on acid soils. In the field notes examined, oaks, pine, hickory, gum, and ash were recorded as the dominant tree species.

Induced pluripotent stem (iPS) cells that are potentially similar to embryonic stem (ES) cells can be artificially established by introduction into somatic cells of the transgenes POU5F1 (also known as Oct3/4), SOX2, KLF4, and c-MYC. In cynomolgus monkeys (Macaca fascicularis), iPS cells generated by using these four allogeneic transgenes should be an important resource for various types of biomedical research because the use of xenogeneic transgenes may cause complications. To establish such iPS cells, cynomolgus monkey somatic cells were infected with amphotropic retroviral vectors, which were derived from Plat-A cells, containing cDNA for the cynomolgus monkey genes POU5F1, SOX2, KLF4, and c-MYC. As a result, iPS cells could be established from somatic cells from fetal liver and newborn skin of cynomolgus monkeys, similarly to the case for mouse and human somatic cells.

The thermodynamics of H(2) solution and hydride formation/decomposition have been determined by reaction calorimetry (303 K) for disordered face centered cubic (fcc) Pd-Mn alloys. This alloy system belongs to the expanded lattice category which predicts that and DeltaH(plat) for H(2) absorption should be more exothermic than those for Pd; the experimental results are that the former is more exothermic, at least at the higher Mn contents, but the latter is not. There is a regular decrease in the H capacity (at p(H)2 = 0.2 MPa) with atom fraction Mn. A linear dependence of log p(H)2 upon H content is found in the single hydride phase for all of these alloys suggesting that DeltaH(H) and DeltaS(H) are also linear functions of r in this region. This is confirmed using the Pd(0.875)Mn(0.125) alloy which has no two-phase region (303 K). It is shown for the Pd(0.875)Mn(0.125) alloy and for Pd that the changes of partial enthalpies and entropies with H content are correlated so as to minimize changes of mu(H).

We discovered two well-demarcated nodules incidentally in a thyroid removed because of a nodular goiter. Histologically, the nodules showed a pattern of paraganglioma or so-called paraganglioma-like adenoma of the thyroid (PLAT), with lobules of polygonal and oval cells in a vascular stroma, but the immunohistochemical markers typical of paraganglioma, including chromogranin, synaptophysin, Leu 7 and 5-100, and thyroglobulin, characteristic of PLAT, were negative in the tumor cells. C-cell markers calcitonin and somatostatin were also negative. Stain for neuron-specific enolase (NSE), however, showed a distinctive pattern of reactivity within cells at the periphery of the lobules, whereas all tumor cells stained positively for keratins. Stain for carcinoembryonic antigen showed a focal interstitial pattern that corresponded to small intercellular spaces filled by microvilli identified ultrastructurally. This pattern of immunohistochemical staining has not been previously described in paraganglioma or in PLAT, and may have implications about the origin and nature of these controversial entities.

Blood smaples were collected in cold O.1 M sodium citrate from 30 healthy young persons whose mandibular impacted molars were to be removed. Blood sampling was repeated immediately after operation. Fibrinolytic activity of the euglobin fraction, precipitated at pH 5.9, was measured by the fibrin plate method. Three different fibrinogen solutions were used: 1) bovine fibrinogen, 2) bovine plasminogen-free fibrinogen, and 3) human fibrinogen. The differences between preoperative and postoperative fibrinolytic activity were evaluated. No increase of fibrinolytic activity was seen in general, indicating that oral surgery does not necessarily influence the fibrinolytic activity of the blood. Dividing the patients according to sex or to a long/short surgical procedure did not influence the activity either. However, in four patients who developed alveolitis sicca (dry socket), a significant rise of activity on all the fibrin plats was seen (P less than 0.01) when compared with the variations measured in patients with normal healing. The result suggests that a rise of blood fibrinolytic activity of a certain magnitude during oral surgery may predispose to the devlopement of alveolitis sicca. Four females who took oral contraceptives showed a higher postoperative fibrinolytic activity (P less than 0.05) than the other females. The importance of this observation is to be investigated in further studies.

The chloroform and alcohol extracts of e plats were screened against six bacteria viz staphylococcus aureus, Escherichia Coli, proteus mirablis, salmonella typhimurium, Pseudomonas aeruginosa and Klebsiella a aerogenes. Benzene-ethyl acetate (4:1) eluate of chloroform extract sowed activity against E.Coli, S.typhimurium and P.aeruginosa. The nbutanol extract of alcohol extract was active against P. mirablis and S. typhimurium. A phenolic compound isolated fro the above fraction exhibited activity against P. mirablis only.