Salivary proteins from dysplastic leukoplakia and oral squamous cell carcinoma and their potential for early detection.

Dysplastic leukoplakia (LP) of the oral cavity is a potentially malignant condition for oral squamous cell carcinoma (OSCC), early detection of which remains an unmet clinical need. In an effort to develop non-invasive biomarker based method for early detection of the disease, differential proteomic profiling was carried out with the saliva from patients with risk habits and diagnosed with LP and those with lymph node negative and positive OSCC in comparison to healthy controls with risk habits. Ninety three proteins were observed at elevated level (≥1.5 fold), and 30 were prioritized based on a scoring system comprising of confidence of identification, presence in the various specimen groups, functional relevance, and their secretory potential. Verification was carried out in independent patient cohorts for 8 selected, representative, upregulated proteins using ELISA. Three of them CD44, S100A7, and S100P were significantly altered in patients with LP as well as OSCC and can be regarded as a panel of biomarker candidates for early detection of the malignancy. Other members may also be investigated in a targeted manner to expand the portfolio of biomarkers for early detection. The mass spectrometry data are available via ProteomeXchange with identifier PXD015722. SIGNIFICANCE: There is an unmet clinical need for non-invasive, biomarker based methods for the improved early detection and the subsequent management of oral cancer. The study represents differential proteome profiling of the saliva of patients with oral dysplastic leukoplakia (LP) - a potentially malignant lesion, patients diagnosed with oral squamous cell carcinoma (OSCC), and healthy controls to identify potential markers for the purpose of early detection of malignancy. From among the matched and prioritized proteins with elevated levels in the saliva of patients with LP and those with OSCC, eight were verified. Three of them - CD44, S100A7 and S100P appeared promising candidates as biomarkers for early detection of the neoplastic predisposition and may form the basis of clinical assays for this purpose.

Data-Driven Discovery of Extravasation Pathway in Circulating Tumor Cells.

Circulating tumor cells (CTCs) play a crucial role in cancer dissemination and provide a promising source of blood-based markers. Understanding the spectrum of transcriptional profiles of CTCs and their corresponding regulatory mechanisms will allow for a more robust analysis of CTC phenotypes. The current challenge in CTC research is the acquisition of useful clinical information from the multitude of high-throughput studies. To gain a deeper understanding of CTC heterogeneity and identify genes, pathways and processes that are consistently affected across tumors, we mined the literature for gene expression profiles in CTCs. Through in silico analysis and the integration of CTC-specific genes, we found highly significant biological mechanisms and regulatory processes acting in CTCs across various cancers, with a particular enrichment of the leukocyte extravasation pathway. This pathway appears to play a pivotal role in the migration of CTCs to distant metastatic sites. We find that CTCs from multiple cancers express both epithelial and mesenchymal markers in varying amounts, which is suggestive of dynamic and hybrid states along the epithelial-mesenchymal transition (EMT) spectrum. Targeting the specific molecular nodes to monitor disease and therapeutic control of CTCs in real time will likely improve the clinical management of cancer progression and metastases.

Virus-induced gene silencing and its application in characterizing genes involved in water-deficit-stress tolerance.

Understanding post-transcriptional gene silencing (PTGS) phenomena in plants has provided breakthroughs in advancing plant functional genomics. A recently developed approach based on one of the strategies adopted by plants to defend against viruses, called virus-induced gene silencing (VIGS), is being widely used to enumerate the function of plant genes. Since its discovery, VIGS has been widely used to characterize plant genes involved in metabolic pathways, homeostasis, basic cellular functions, plant-microbe, plant-nematode and plant-herbivore interaction. Recently, the application of this technique has been extended to characterize the genes and cellular processes involved in abiotic-stress tolerance, and in particular drought and oxidative stress. Because abiotic-stress tolerance is multigenic, identification and characterization of genes involved in this process is challenging. VIGS could become one among the several potential tools in understanding the relevance of these stress-responsive genes. Development of VIGS protocols for the use of heterologous gene sequences as VIGS-inducers has extended its applicability to analyze genes of VIGS recalcitrant plant species. This article describes the methodology of VIGS for characterizing the water-deficit-stress-responsive genes, precautions to be taken during the experimentation, and future application of this technology as a fast forwarded as well as a reverse genetics tool to identify and characterize plant genes involved in drought tolerance. We also describe the importance of accurate water-deficit-stress imposition and quantification of stress-induced changes in the silenced plants during the process of screening to identify genes responsible for tolerance. Further, limitations of VIGS in characterizing the abiotic-stress-responsive genes are noted, with suggestions to overcome these limitations.

Nutritional and hematological impact of dietary flaxseed and defatted flaxseed meal in rats.

An 8-week study was conducted to determine the impact of dietary ground flaxseed (FS) or defatted flaxseed meal (FLM) on plasma lipids, minerals, hematological parameters and vitamin E status of weanling female Sprague-Dawley rats. These rats were fed isocaloric modified AIN-76 diets supplemented with 0.0, 5.0, 10.0% (w/w) FS or 6.2% (w/w) FLM for 56 days. Total and HDL cholesterol were not influenced by any of the dietary treatments. Plasma triglyceride was significantly increased by FLM, but not affected by FS. Total RBC counts and hematocrit were significantly higher in FS groups than in the control group; however, hemoglobin was not affected by FS. Dietary FLM had no effect on any of the above hematological parameters. Plasma alkaline phosphatase, an indicator of Zn status and a marker of bone formation, was significantly lower in the FS and FLM groups than in the control group. Plasma vitamin E content was not influenced by dietary treatment. Liver vitamin E was significantly higher in groups fed 10% FS and 6.2% FLM. In summary, moderate amounts of dietary FS may have the potential to increase liver vitamin E level and improve iron status. However, FS/FLM consumption may have a negative effect on zinc status, as indicated by decreased alkaline phosphatase levels.

Application of Azure C for the extractive spectrophotometric determination of microgram amounts of penicillin.

A simple, accurate, and rapid method for the quantitative determination of penicillin is proposed. The method is based on the formation of a blue penicillin-Azure-C ion-pair that can be extracted into chloroform in phosphate-citric acid buffer. The molar absorptivities for sodium penicillin G and potassium penicillin V at 635 nm were 5.46 X 10(3) and 2.19 X 10(4) l/mol/cm, respectively. Beer's law was valid over the concentration range of 4-80 micrograms/ml for sodium penicillin G and 3-55 micrograms/ml for potassium penicillin v. Maximum absorbance was obtained almost instantaneously and was stable for several days. The method was successfully applied to pharmaceutical preparations.

Potentiometric and visual titrations with bromamine-B.

Bromamine-B is proposed as an oxidimetric titrant for potentiometric and visual end-point titrations of arsenic(II), hexacyanoferrate(II), antimony(III), hydroquinone, semicarbazide hydrochloride, isonicotinic acid hydrazide, hydrazine sulphate, ascorbic acid, phenylhydrazine hydrochloride and metol. Quinoline Yellow, naphthidine, dimethylnaphthidinedisulphonic acid, o-dianisidine, diphenylbenzidine, Variamine Blue, alpha-naphthoflavone, Amaranth, Methyl Orange and Methyl Red are proposed as indicators in macro and micro titrations of the reductants with bromamine-B. The transition potentials of Quinoline Yellow, naphthidine, dimethylnaphthidinedisulphonic acid, and o-dianisidine in the titration of ascorbic acid are reported. Arsenic(III) and hexacyanoferrate(II) are suggested for the standardization of bromamine-B solutions.

Redox indicators in titrations with dichloramine-B.

Naphthidine, dimethylnaphthidine, dimethylnaphthidinedisulphonic acid, o-dianisidine, Quinoline Yellow, diphenylbenzidine and Amaranth are proposed as indicators in titrations of arsenic(III), iron(II), antimony(III), hydroquinone, hydrazinium sulphate, phenylhydrazine hydrochloride, semicarbazide hydrochloride and ascorbic acid with dichloramine-B. They give a very sharp colour change at the equivalence point. Arsenic(III) and iron(II) are suggested for standardization of dichloramine-B solutions. A potentiometric method for the determination of arsenic(III) and semicarbazide hydrochloride is described.

N-substituted phenothiazines as redox indicators in titrations with chloramine-T and chloramine-B.

Profenamine hydrochloride, fluphenazine dihydrochloride, trifluopromazine hydrochloride, cyamepromazine maleate, perphenazine dihydrochloride and mepazine hydrochloride are proposed as redox indicators in the titration of hydroquinone, metol and ascorbic acid with chloramine-T and chloramine-B in sulphuric, hydrochloric and acetic acid media. They give a sharp reversible colour change at the equivalence point. A simple but accurate method for the determination of hydroquinone, metol and ascorbic acid is described. The conditional potentials and molar absorptivities of the indicators and redox potential of chloramine-B are reported.

N-substituted phenothiazines as redox indicators in bromatometry.

Diethazine hydrochloride, butaperazine dimaleate, trifluoperazine hydrochloride, promethazine hydrochloride, prochlorperazine maleate and chlorpromazine hydrochloride have been studied as indicators in bromate titration of quinol, metol and ascorbic acid. They give a very sharp reversible colour change at the equivalence point. Their formal potentials have been determined. A simple but accurate method for the estimation of quinol and metol is reported.

Influenza in a geriatric unit.

An outbreak of influenza A (resembling A/Victoria/3/75) occurred in 2 geriatric wards in February 1976 and, out of 19 patients, 3 died. Of the patients who were at risk, 30% were affected. Immunization for influenza in the elderly just before the winter is suggested. The procedure is simple and inexpensive. The recommendations of the DHSS (U.K.) for the use of influenza vaccine to practising doctors is, therefore, welcome. The suitable treatment appears to be that of repeated immunization with inactivated vaccine. Amantadine in the elderly population may also be effective, and is worthy of trial.

Rapid spectrophotometric determination of platinum with trifluoperazine dihydrochloride.

Trifluoperazine dihydrochloride reacts with platinum(IV) in sodium acetate-hydrochloric acid buffer containing copper(II) catalyst to form a bluish-green 1:1 complex with absorbance maximum at 504 nm. A 50-fold ratio of reagent to metal ion is necessary for complete complexation. Beer's law is valid over the concentration range 0.5-8 ppm of platinum(IV) with optimum concentration range 1-7 ppm. The molar absorptivity is 6.50 x 10(3) l.mole(-1).cm(-1). The effects of pH, time, temperature, concentration of reagent and copper, order of addition of reagents, and the interferences from various ions are reported.

Rapid spectrophotometric determination of ruthenium with diethazine hydrochloride.

Diethazine hydrochloride reacts with ruthenium(III) instantaneously at high acidity (sulphuric or hydrochloric acid) to form a red 1:1 complex with absorbance maximum at 515nm. A tenfold molar excess of reagent is necessary for complete complexation. Beer's law is obeyed for 1.0-10.4 ppm of ruthenium(III), with optimum concentration range 2-9 ppm. The molar absorptivity is 5.35 x 10(3) 1.mol(-1) .cm(-1).

Effect of different glycerol levels on the release of glutamic oxaloacetic transaminase (GOT) from deep frozen ram spermatozoa.

Release of GOT was determined during different treatments in ram semen extended (1:10) with lactose-egg yolk diluter containing 3.5% and 5% glycerol (v/v). Semen samples were collected by the electro-ejaculation technique and GOT was analysed colorimetrically. Highly significant difference (p less than 0.01) was found in the post-thaw percent motile spermatozoa and GOT release (IU/10(9) spermatozoa) between 3.5% and 5% glycerol containing equilibrated samples. The difference in percent motile spermatozoa was not significant between 3.5% and 5% glycerol levels in control, post-thaw unequilibrated and prefreeze equilibrated samples. Whereas the difference in GOT release between 3.5% and 5% glycerol concentrations was not significant in control and post-thaw unequilibrated samples and highly significant (p less than 0.01) in prefreeze equilibrated samples.