Inter- and intra-rater reliability and agreement in determining subcutaneous tumour margins in dogs.

The objective of this prospective study was to evaluate agreement and reliability of calliper-based measurements of locally invasive subcutaneous malignant tumours in dogs. Four raters measured the longest diameter of 12 subcutaneous tumours (7 soft tissue sarcomas and 5 mast cell tumours) from 11 client-owned dogs during 3 randomized, blinded measurement trials, both pre- and post-sedation. Inter- and intra-rater reliability was evaluated using intra-class correlation coefficient (ICC) and agreement was evaluated using Bland-Altman plots. Inter- and intra-rater reliability was good (ICC range of 0.8694-0.89520) and excellent (ICC range of 0.9720-0.9966), respectively. For agreement calculations, an a priori clinically relevant limit of agreement of 10 mm was set. Inter- and intra-rater agreement was unacceptable with inter-rater limits of agreement ranging from 15.9 to 55.6 mm and intra-rater limit of agreement ranging from 11.9 to 28.1 mm. Review of the measurement trial photographs revealed that calliper orientation changes were frequent, occurring in 9/12 (75%) and 8/12 (67%) pre- and post-sedation cases. No significant correlation was found between inter-rater measurement standard deviations and calliper orientation changes or dog body condition score. These findings suggest veterinarians may have poor agreement in determining the gross edge of tumours, which is expected to introduce bias and inconsistency in tumour staging, assessing response to therapy, and surgical margin planning. Due to the potential consequences for veterinary cancer patients, future studies are needed to validate the present findings.

Variations in single/two stage thyroidectomies for cancer may be due to differences in thyroid fine needle cytology provision.

BACKGROUND & AIMS: Recommended treatment for thyroid cancers >10 mm is single stage total thyroidectomy (SST). Cancers diagnosed by diagnostic lobectomy may need completion surgery resulting in two stage thyroidectomies (TST). We noticed significant variation in numbers of SST and TST between hospitals within our cancer network and explored reasons for this using a prospective database containing all cases from 2004 to 2011 (n = 1030). We therefore conducted a survey of thyroid cytology provision across the network during 2010-2011. METHODS: A central university hospital with the largest caseload (21.5% of total) was chosen as "benchmark". Of 14 remaining hospitals 3 were excluded from analysis due to low thyroid operation numbers and the remaining compared with benchmark. We used individual chi-squared tests with Bonferroni correction to explore variation in expected and observed numbers of SST/TST. Analysis of variance (ANOVA) was used to examine reasons for observed differences. RESULTS: Significant variance in SST/TST was seen between hospitals (p < 0.00001). Three hospitals had frequencies of SST statistically similar to reference hospital; each reported 201-300 thyroid cytology cases during the survey period. The remaining 8 had lower rates of SST, the 2 lowest performing hospitals having SST rates of 11% (p = 0.0004) and 9% (p < 0.0001). These eight hospitals reported fewer than 200 cytology cases each, shared amongst 4-7 pathologists per site. Differences were unrelated to patient age, gender, tumour histology or stage (ANOVA). Only the reference hospital had specialist cytopathologists. CONCLUSION: Variation in thyroid cytology provision may increase TST rates. Thyroid cytology should be concentrated in high volume centres with specialist thyroid cytopathologists.

Determination of Trace Metals, Moisture, pH and Assessment of Potential Toxicity of Selected Smokeless Tobacco Products.

The characterization and classification of smokeless tobacco products has been a continuously evolving process. This is based on a number of different parameters like nicotine content, moisture content, amount of heavy metals, pH, and in vitro cytotoxicity assays. Their contexts often vary between countries, research institutions, and legal requirements. The categorisation of these products is quite challenging due to the diffused sample sizes, diverse array of branded products on offer, and the absence of a centralized manufacturing facility. This study aims at a systematic classification of 10 smokeless tobacco product samples from the retail market based on their potential toxicity upon long-term use. The estimation of potential toxicity follows a well-established method that employs the concentration of toxic metals in the different samples. The potential toxicity as well as heavy metal concentrations of the smokeless tobacco products analysed was found to be much higher than acceptable limits. For instance, the levels of lead, cadmium, copper and zinc of 2.5, 1, 4 and 23 ppm, respectively, are well above their recommended limits. The results from the study indicate that chronic use of smokeless tobacco products is a significant health risk, especially in the vulnerable population. Further studies of this nature will help establish a toxicological fingerprint on the diverse class of products that floods the market now.

Bioremoval of Basic Violet 3 and Acid Blue 93 by Pseudomonas putida and its adsorption isotherms and kinetics.

Basic Violet 3 and Acid Blue 93 are the most important group of synthetic colourants extensively used in textile industries for dyeing cotton, wool, silk and nylon. Release of these dye pollutants in to the environment adversely affects the human health and aquatic organisms. The present study we used Pseudomonas putida MTCC 4910 for the adsorptive removal of Basic Violet 3 and Acid Blue 93 from the aqueous solutions. The pH (4-9) and NaCl concentrations (1mM-1M) did not influence the adsorption process. The equilibrium adsorption process fitted well to Freundlich model than Langmuir model. The kinetics of adsorption fitted well by pseudo-second-order. Thus in the present study an attempt has been made to exploit the dye removal capability of P. putida MTCC 4910, and it was found to be an efficient microbe that could be used for bio removal of dyes from textile effluents.

Validation of self-reported hearing loss using television volume.

OBJECTIVE: To assess the diagnostic utility of using television volume as a marker for hearing loss. STUDY DESIGN: Prospective study using a self-administered questionnaire. SETTING: ENT and audiology out-patient departments in the north of England. PARTICIPANTS: One hundred and seventeen patients with a history of hearing loss, undergoing pure tone audiometry for the first time. MAIN OUTCOME MEASURES: sensitivity, specificity, diagnostic accuracy, and positive and negative predictive value of television volume as a marker of hearing loss. RESULTS: The data indicated that if the patient (or their partner or parent) reported viewing television with an increased volume, then there was a 68 per cent chance of the patient having a hearing loss of 25 dB or more. Patients reporting increased television volume had a mean hearing loss of 35 dB. Increased television volume had a sensitivity of 81 per cent and a specificity of 52 per cent as a predictor of hearing loss. Patients who increased their television volume to watch news programmes had an average hearing loss of 41 dB; increased television volume for news programmes had a sensitivity of 75 per cent and a specificity of 71 per cent as a predictor of hearing loss. CONCLUSIONS: Television volume is a useful marker of hearing loss in situations where audiometry is unavailable, for instance in a primary care setting. However, it is not a very specific test.

Skin incisions for the excision of spherical cutaneous and subcutaneous lesions.

Spherical cutaneous and subcutaneous lesions such as sebaceous cysts are often removed via an ellipse, particularly if they are relatively large. Deciding the width of the ellipse can be difficult. We calculated that if the width of the ellipse is one-third of the half circumference, then the wound should close satisfactorily.

A cytotoxic substance produced by a wild culture of Lactobacillus casei D-34 against tumour cells.

A wild lactic culture isolated from dahi (fermented milk) sample and characterised as L. casei D-34 was found to be significantly cytotoxic (34-36%) against three tumour cell lines, HeLa, HEp-2 and HFS-9. The cytotoxic substance (CS) was found to be in the culture supernatants, protein in nature, with a molecular weight ranging from 17,000-20,000. The crude culture supernatant was partially purified by dialysis and ion exchange chromatography as anionic, cationic and neutral fractions. Among the fractions, except for the anionic fraction, others were found to be highly cytotoxic against all three tumour cell lines. The cationic, neutral and pooled (anionic:cationic:neutral in 1:1:1 ratio) fractions showed 50, 70, 70% cytotoxicity against Hep-2 cells, 70, 88, 94% against HFS-9 cells and 50, 89, 90% against HeLa cells respectively. Pooled fraction was found to exhibit higher percent of cytotoxicity compared to individual fractions indicating a synergistic effect. (3H)-thymidine incorporation studies revealed that CS and its fractions inhibited DNA synthesis in tumour cells. The CS was stable towards heat and pH changes.

Cytotoxic activity of lactobacilli isolated from plant and dairy sources.

Forty four cultures of Lactobacilli isolated from their natural sources such as dahi, raw milk and fermenting rice-pulse doughs etc. along with four standard strains of Lactobacilli were assayed for their cytotoxic activity against three secondary tumour cell lines. Three cultures isolated from dahi samples and identified as Lactobacillus casei D-34, L. casei D-48a, L. plantarum D-70a along with one standard strain L. casei B 1922 exhibited significant cytotoxic activity in the range of 30 to 36%.

Changes in enterococcal population during storage of reconstituted infant food.

A six-fold increase in the enterococcal population was observed in reconstituted infant food samples after storage for 2 h at 37 degrees C. The increase in enterococcal counts at 40 degrees C and 45 degrees C was approximately five-fold during the same period. However, the corresponding total viable counts increased by twelve fold at these temperatures after 2 h. After 12 h, the enterococcal and total viable counts increased to 39 x 10(4) and 36 X 10(7) colony forming units per ml, at 37 degrees C respectively. A similar pattern in enterococcal and total bacterial count was observed at 40 degrees and 45 degrees C. TNase was detected in reconstituted infant food samples held at 37 degrees, 40 degrees and 45 degrees C, after 12 h, while pH values declined to 5.0, 5.1 and 5.2, respectively at the above temperatures. From TNase positive samples, an isolate S. faecium IF-100 capable to produce TNase was recovered. Storage of reconstituted infant food samples in the refrigerator (5 degrees C) resulted in a gradual increase in enterococcal population which reached 39 X 10(3) c.f.u. per ml after 12 days. However, TNase was not detected in any of these samples.

Enterocin typing of enterococci isolated from dried infant foods.

One hundred and fifty enterococcal isolates recovered from 16 market samples of infant foods and 35 from other sources were characterized and subjected to enterocin typing with 18 indicator strains. Among 150 enterococcal isolates, 114 (76%) were able to be typed by the indicator strains. Although 24 enterocin patterns were observed with these enterococci, the most prevalent types were X-9, 224, and 65-603. Occurrence of pattern X-9 either singly or in combination with many other types was most frequent. Many of the enterocin patterns in enterococcal isolates were recovered from samples of dairy water supply and hand washings of personnel working in a dairy plant that manufactured infant food; this suggests the possibility of these as sources of contamination. Enterocin typing of enterococci could prove useful in epidemiological studies.

Toxigenic Bacillus cereus var. fluorescens in human milk.

Human milk with high bacterial load of B. cereus var. fluorescens is reported. The organism was identified by morphological, cultural, and biochemical characteristics. Lecithinase and haemolysin were produced in rabbit and bovine blood. The toxigenic nature of the organism was confirmed by production of necrosis on rabbit skin and fluid accumulation in rabbit ileal loop. An antibiotic resistogram also has been included.

Preliminary Studies on Antimicrobial Activity of Streptococcus lactis subsp. diacetylactis.

Streptococcus lactis subsp. diacetylactis is well-known for its ability to inhibit pathogenic and milk spoilage microorganisms. In this study, an attempt was made to standardize the method for estimation of antimicrobial activity of the organism. Fourteen strains of S. lactis subsp. diacetylactis were examined for production of antimicrobial substances and all strains were found to produce antimicrobial substances. Of the strains, strain DRC1 produced maximum amounts of antimicrobial substances followed by strains DRC2 and S1. Mutants of S1 (S1-67/C and S1-195) produced higher amounts of antimicrobial substances than all other strains tested. The antimicrobial properties of strains S1-67/C were assessed against 28 bacterial and mold cultures. Cell-free filtrates of S1-67/C strain were more inhibitory to gram-negative than gram-positive bacteria. This organism also inhibited the growth of Geotrichum candidum but failed to inhibit other molds tested in this study.

Nutritional Factors Affecting Growth and Production of Antimicrobial Substances by Streptococcus lactis subsp. diacetylactis S1-67/C.

The present study pertains to the effect of nutritional factors on the growth and production of antimicrobial substances (AS) by Streptococcus lactis subsp. diacetylactis S1-67/C. Among nine media tested, yeast extract dextrose broth supported good growth and maximum production of AS. Addition of beef extract and yeast extract at 1.0 and 0.6% levels, respectively, increased growth as well as production of AS. Of ten carbohydrates examined, maximum production of AS was achieved with 1% glucose followed by fructose, 4% molasses, lactose, sucrose, galactose, mannitol, maltose and 2% molasses. Xylose inhibited production of AS, although it stimulated growth of the organism. Peptone, tryptone and tryptose (each at the 1.5% level) significantly stimulated production of AS. Other nitrogen sources, including soytone, casein hydrolysate and proteose peptone, retarded production of inhibitory substances. Among the amino acids, L-leucine, DL-methionine and L-glutamic acid were most essential for growth and production of AS, whereas L-lysine, L-proline, DL-serine, DL-asparatic acid, L-arginine-HCl and DL-tryptophan were stimulatory. Other amino acids such as DL-ornithine, L-cysteine-HCl and DL-citrulline slightly stimulated AS production. In the presence of cynocobalmin, niacin, folic acid, calcium pantothenate and riboflavin, S. lactis subsp. diacetylactis S1-67/C produced maximum amounts of inhibitory substances. Omission of individual mineral salts from the basal medium did not affect production of AS by the organism.

Characterization of Deoxyribonuclease-Positive Enterococci Isolated from Milk and Milk Products.

Five hundred and ten isolates of enterococci recovered from milk and milk products were screened for deoxyribonuclease (DNase) production. Of the 166 (32.5%) DNase-positive cultures, 29 (5.7%) produced thermonuclease that resisted boiling for 15 min. Although the incidence of thermolabile DNase-positive enterococci was 50.5% in Cheddar cheese, the majority of thermonuclease-producing enterococci was recovered from dried milks and infant foods. On the basis of biochemical, physiological and serological tests, all DNase-producing enterococci were characterized as S. faecalis var. faecalis (21), S. faecalis var. zymogenes (9), S. faecalis var. liquefaciens (90), S. faecium (23) and S. durans (16). The predominant thermonuclease-positive type was S. faecalis var. faecalis followed by S. faecalis var. zymogenes . Six strains of enterococci were found to be toxigenic when tested in ligated rabbit ileal loops, infant mice and rabbit skin.

Aflatoxin Production in Khoa.

Aspergillus parasiticus NRRL 2999 and Aspergillus flavus K3 were examined for aflatoxin production in khoa. The inoculated khoa samples were examined after incubation for 2 weeks at 5, 28 and 37 C. Aflatoxin penetrated to a depth of 4 cm in the inoculated khoa samples stored at 28 and 37 C.

Factors Affecting Lipase Production by Mucor racemosus.

Mucor racemosus isolated from butter exhibited maximum lipase production at 22 C in 3 days at pH 5.0, when grown as a static culture. Supplementation of the medium with glucose and lactose at 1% level resulted in maximum (18.0 µmoles FFA) and minimum (10.5 µmoles FFA) enzyme production. Of the five different nitrogen sources tested, peptone at the 2% level supported highest production of lipase. Calcium, potassium and sodium citrate (0.1 %), when incorporated in the medium, stimulated production of enzyme to the extent of 80, 60 and 47%, respectively.