Stool sample storage conditions for the preservation of Giardia intestinalis DNA

Stool is chemically complex and the extraction of DNA from stool samples is extremely difficult. Haemoglobin breakdown products, such as bilirubin, bile acids and mineral ions, that are present in the stool samples, can inhibit DNA amplification and cause molecular assays to produce false-negative results. Therefore, stool storage conditions are highly important for the diagnosis of intestinal parasites and other microorganisms through molecular approaches. In the current study, stool samples that were positive for Giardia intestinalis were collected from five different patients. Each sample was stored using one out of six different storage conditions [room temperature (RT), +4ºC, -20ºC, 70% alcohol, 10% formaldehyde or 2.5% potassium dichromate] for DNA extraction procedures at one, two, three and four weeks. A modified QIAamp Stool Mini Kit procedure was used to isolate the DNA from stored samples. After DNA isolation, polymerase chain reaction (PCR) amplification was performed using primers that target the β-giardin gene. A G. intestinalis-specific 384 bp band was obtained from all of the cyst-containing stool samples that were stored at RT, +4ºC and -20ºC and in 70% alcohol and 2.5% potassium dichromate; however, this band was not produced by samples that had been stored in 10% formaldehyde. Moreover, for the stool samples containing trophozoites, the same G. intestinalis-specific band was only obtained from the samples that were stored in 2.5% potassium dichromate for up to one month. As a result, it appears evident that the most suitable storage condition for stool samples to permit the isolation of G. intestinalis DNA is in 2.5% potassium dichromate; under these conditions, stool samples may be stored for one month.

Optimization of the carrot leaf dehydration aiming at the preservation of omega-3 fatty acids

The carrot leaf dehydration conditions in air circulation oven were optimized through response surface methodology (RSM) for minimizing the degradation of polyunsaturated fatty acids, particularly alpha-linolenic (LNA, 18:3n-3). The optimized leaf drying time and temperature were 43 h and 70 ºC, respectively. The fatty acids (FA) were investigated using gas chromatography equipped with a flame ionization detector and fused silica capillary column; FA were identified with standards and based on equivalent-chain-length. LNA and other FA were quantified against C21:0 internal standard. After dehydration, the amount of LNA, quantified in mg/100 g dry matter of dehydrated carrot leaves, were 984 mg.

Efficacy of varicose vein surgery with preservation of the great safenous vein

OBJECTIVE: To evaluate the efficacy of surgical treatment of varicose veins with preservation of the great saphenous vein. METHODS: We conducted a prospective study of 15 female patients between 25 and 55 years of age with clinical, etiologic, anatomic and pathophysiologic (CEAP) classification 2, 3 and 4. The patients underwent surgical treatment of primary varicose veins with great saphenous vein (GSV) preservation. Doppler ultrasonography exams were carried out in the first and third months postoperatively. The form of clinical severity of venous disease, Venous Clinical Severity Score (VCSS) was completed before and after surgery. We excluded patients with history of deep vein thrombosis, smoking or postoperatively use of elastic stockings or phlebotonics. RESULTS: All patients had improved VCSS (p <0.001) and reduction in the diameter of the great saphenous vein (p <0.001). There was a relationship between VCSS and the GSV caliber, as well as with preoperative CEAP. There was improvement in CEAP class in nine patients when compared with the preoperative period (p <0.001). CONCLUSION: The varicose vein surgery with preservation of the great saphenous vein had beneficial effects to the GSV itself, with decreasing caliber, and to the symptoms when the vein had maximum caliber of 7.5 mm, correlating directly with the CEAP. The decrease in GSV caliber, even without complete abolition of reflux, leads to clinical improvement by decreasing the reflux volume.

Heat shock and salicylic acid on postharvest preservation of organic strawberries

Heat shock and salicylic acid have been studied on shelf-life extension of fruits. The benefits of these techniques have been related to their effect on inducing physiological defense responses against the oxidative stress and pathogen development. The objective of this study was to evaluate the effect of heat shock and salicylic acid on the postharvest preservation and contents of total phenolics, anthocyanins, ascorbic acid, fresh weight loss and microbiological quality of organic strawberries cv. Dover. Strawberries produced organically and stored at 5 ºC were subjected to heat shock (45 ºC ± 3 ºC for 3 h), application of salicylic acid (soaking in 2.0 mmol L-1 solution), heat shock in combination with salicylic acid and control. After treatment, the fruits were packed and stored in a climatic chamber at 5 ºC ± 2 ºC. At 1, 7 and 14 days, the experimental units were removed from refrigeration and kept at room temperature of approximately 20 ºC for two days. There was no effect of treatments on fresh weight loss, incidence of pathogens or chemical variations in strawberry fruits during the storage period. In natural conditions, organically grown strawberries remained in good condition for sale up to seven days of storage in all treatments.

Preservation of rhizobia by lyophilization with trehalose

The objective of this study was to determine if trehalose can effectively enhance the viability and storage lifetime of Rhizobium tropici and Rhizobium etli after lyophilization as compared to the traditional protective combination of sucrose and peptone. Two strains of bean-nodulating rhizobia were effectively preserved in the lyophilized state by 100 mM trehalose, and survived for at least 12 days, even when stored under non-ideal conditions. Trehalose provided better protection than the sucrose/peptone combination. When protective excipients were introduced inside the cells, strain CFN 42 was more sensitive to the type of additive used than strain CIAT 899, probably because CIAT 899 produces beta (1-2) glucan, which may have a protective effect. beta (1-2) glucan combined with sucrose protected better than trehalose against leakage from freeze-dried liposomes, but by itself it had no protective capability. There was a high level of unfrozen water associated with the hydrated glucan, in addition to a thermal event at approximately 70ºC, corresponding to an observed gel-sol transition in the glucan. These findings may shed light on the mechanism by which the glucan contributes to the protection of dried cells and liposomes, and may lead to further improvements in rhizobial inoculants.

Preservation of minimally processed 'aurora-1' peaches using additives

'Aurora-1' peaches establishes an interesting alternative as a minimally processed product, due to its characteristics like flavor, color, smell, and also because of its handling resistance. However, it has a short shelf life after a fresh-cut due to enzymatic browning and stone cavity collapse. The main purpose of this research was to test the additive with antioxidant effect to prevent browning in minimally processed 'Aurora-1' peaches. The minimal processing consists of washing, sanitizing, peelings and fruit stone extraction. After that, longitudinal cuts were made to obtain eight segments per fruit. The slices were immersed into the following treatment solutions: control (immersion in 2% ascorbic acid); 2% ascorbic acid + 2% calcium chloride; 1% sodium isoascorbate; 1% citric acid; 1% L-cysteine hydrochloride. The products were placed into rigid polystyrene trays branded MEIWA M-54, covered with 14 µm PVC film (OmnifilmTM) and kept in cold storage at 3ºC ± 2ºC and 65% RH for twelve days, and evaluated each three days. Appraised variables were appearance, soluble solids, titratable acidity, soluble carbohydrates and reducing sugars, total and soluble pectin, ascorbic acid, and peroxidase and polyphenol oxidase enzyme activity. L-cysteine gave to the minimally processed products a shelf life of twelve days, limmited by off-flavor. The treatment with ascorbic acid was efficient to maintainthe ascorbic acid content, with a shelf-life of nine days, limited by enzymatic browning.

PRESERVATION OF PRIMARY FOREST CHARACTERISTICS DESPITE FRAGMENTATION AND ISOLATION IN A FOREST REMNANT FROM VIÇOSA, MG, BRAZIL1

According to its owners, the Forest of Seu Nico (FSN) from the Viçosa municipality, Minas Gerais, Brazil, never has been logged and is therefore considered a primary forest. Nevertheless, the forest patch suffered impacts due to selective wood and non-timber extraction, fragmentation and isolation. Aim of this study was to test if the FSN, despite impacts, preserved characteristics of primary forests, which are elevated percentages of non-pioneer (>90%), animal-dispersed (>80 %), understory (>50%) and endemic species (~40%). For that, all trees with diameter at breast height equal or major than 3.2 cm within a plot of 100 x 100 m were identified. With 218 tree species found within this hectare, the FSN's species richness is outstanding for the region. The percentages of non-pioneer (92 %), animal-dispersed (85 %), understory (55 %) and endemic species (39.2 %) from the FSN fulfill the criteria proposed for primary forest. Therefore, we conclude that the FSN maintained its characteristics as a primary forest which highlights its importance for the conservation of biotic resources in the region, where similar fragments are lacking or not described yet.

Preservation of graft function in low-risk living kidney transplant recipients treated with a combination of sirolimus and cyclosporine

The use of sirolimus (SRL) in combination with full doses of cyclosporin A (CsA) results in reduced one-year kidney allograft function, which is associated with shorter long-term allograft survival. We determined the effect of reduced CsA exposure on graft function in patients receiving SRL and prednisone. Ninety recipients of living kidney transplants receiving SRL (2 mg/day, po) were compared to 35 recipients receiving azathioprine (AZA, 2 mg kg-1 day-1, po). All patients also received CsA (8-10 mg kg-1 day-1, po) and prednisone (0.5 mg kg-1 day-1). Efficacy end-point was a composite of biopsy-confirmed acute rejection, graft loss, or death at one year. Graft function was measured by creatinine, creatinine clearance, and graft function deterioration between 3 and 12 months (delta1/Cr). CsA concentrations in patients receiving SRL were 26% lower. No differences in one-year composite efficacy end-point were observed comparing SRL and AZA groups (18 vs 20%) or in the incidence of biopsy-proven acute rejection (14.4 and 14.3%). There were no differences in mean ± SD creatinine (1.65 ± 0.46 vs 1.60 ± 0.43 mg/dl, P = 0.48) or calculated creatinine clearances (61 ± 15 vs 62 ± 13 ml/min, P = 0.58) at one year. Mean ± SD delta1/Cr (-11 ± 17 vs -14 ± 15%, P = 0.7) or the percentage of patients with >20% (26 vs 31%, P = 0.6) or >30% delta1/Cr (19 vs 17%, P = 1) did not differ between the two groups. The use of 2-mg fixed oral doses of SRL and reduced CsA exposure was effective in preventing acute rejection and preserving allograft function.

Fuzzy rule-based system for evaluation of uncertainty transaction in cassava chain

A fuzzy ruled-based system was developed in this study and resulted in an index indicating the level of uncertainty related to commercial transactions between cassava growers and their dealers. The fuzzy system was developed based on Transaction Cost Economics approach. The fuzzy system was developed from input variables regarding information sharing between grower and dealer on “Demand/purchase Forecasting”, “Production Forecasting” and “Production Innovation”. The output variable is the level of uncertainty regarding the transaction between seller and buyer agent, which may serve as a system for detecting inefficiencies. Evidences from 27 cassava growers registered in the Regional Development Offices of Tupa and Assis, São Paulo, Brazil, and 48 of their dealers supported the development of the system. The mathematical model indicated that 55% of the growers present a Very High level of uncertainty, 33% present Medium or High. The others present Low or Very Low level of uncertainty. From the model, simulations of external interferences can be implemented in order to improve the degree of uncertainty and, thus, lower transaction costs.

Influence of the use of acids and films in post-harvest lychee conservation

Lychee (Litchi chinensis Sonn.) has a high commercial value; however, it has a short shelf-life because of its rapid pericarp browning. The objective of this study was to evaluate the shelf-life of 'Bengal' lychee fruits stored after treatment with hydrochloric acid and citric acid, associated with cassava starch and plastic packaging. Uniformly red pericarp fruits were submitted to treatments: 1-(immersion in citric acid 100 mM for 5 minutes + cassava starch 30 g L-1 for 5 minutes), 2-(immersion in hydrochloric acid 1 M for 2 minutes + starch cassava 30 g L-1 for 5 minutes), 3-(immersion in citric acid 100 mM for 5 minutes + polyvinyl chloride film (PVC, 14 µm thick)) and 4-(immersion in hydrochloric acid 1 M for 2 minutes + PVC film). During 20 days, the fruits were evaluated for mass loss, pericarp color, pH, soluble solids and titratable acidity, vitamin C of the pulp and pericarp and activities of polyphenol oxidase and peroxidase of the pericarp. The treatment with hydrochloric acid associated with PVC was the most effective in maintaining the red color of the pericarp for a period of 20 days and best preservation of the fruit. The cassava starch associated with citric acid, and hydrochloric acid did not reduce the mass loss and did not prevent the browning of lychee fruit pericarp.

Revisiting the concept of hepatosplenic schistosomiasis and its challenges using traditional and new tools

Different aspects of hepatosplenic schistosomiasis are revisited here. Manson's schistosomiasis causes periportal fibrosis and portal hypertension in approximately 6% of infected subjects, usually with preservation of their hepatic function. The assessment of liver involvement is of major importance in determining the prognosis and risk of complications from schistosomiasis, such as upper digestive bleeding secondary to variceal rupture. For many years, the diagnosis of hepatosplenic schistosomiasis and liver fibrosis was made by abdominal palpation and the finding of liver and/or spleen enlargement. However, there is no consensus regarding the clinical parameters of the liver and spleen to be considered in this physical evaluation. For the last three decades, abdominal ultrasound (US) has become the best imaging technique to evaluate liver fibrosis caused by schistosomiasis mansoni. However, US is a subjective procedure and is therefore examiner-dependent. Magnetic resonance imaging (MRI) findings have provided valuable information in addition to ultrasound and clinical examination. The combination of a comprehensive history and physical examination, basic laboratory tests (a stool examination for Schistosoma mansoni eggs and a blood cell count), biomarkers for liver fibrosis/portal hypertension and imaging methods seem to offer the best approach for evaluating patients with this disease. In situations where research is involved or in patients with severe disease, MRI may be considered.

Species of the subfamily Triatominae Jeannel, 1919 (Hemiptera: Reduviidae) present in the Collection of Chagas Disease Vectors (FIOCRUZ-COLVEC), State of Minas Gerais

Introduction Biological collections are depositories of information on different species and contribute to the knowledge, protection, conservation and maintenance of biodiversity. Methods A list of triatomine species currently included in the Collection of Chagas Disease Vectors (FIOCRUZ-COLVEC) was prepared from the database made available by the Reference Center on Environmental Information. Results COLVEC curatorship houses 4,778 specimens of triatomines, of which 811 come from other American countries (Argentina, Bolivia, Colombia, Costa Rica, the United States of America, Guatemala, Mexico, Peru, Uruguay and Venezuela) and 3,967 are autochthonous from Brazil. Altogether, 56 species of Chagas disease vectors are represented in the COLVEC: two species of the Tribe Cavernicolini Usinger, 1944; fifteen species of the tribe Rhodniini Pinto, 1926, of which 12 are of the genus Rhodnius and 3 are of the genus Psamolestes; and 39 species of the tribe Triatomini Jeannel, 1919, represented by the genus Dipetalogaster, two species of the genus Eratyrus, two of the genus Meccus, seven of the genus Panstrongylus and 27 of the genus Triatoma. Conclusions This list provides important data on the diversity of triatomines currently included in COLVEC, including the expanded area of Panstrongylus lutzi occurrence in the municipalities Pirapora and Januária, State of Minas Gerais. The maintenance and expansion of the collection ensures the preservation of biodiversity and further studies.

Pelvic lymphoscintigraphy: contribution to the preoperative staging of rectal cancer

PURPOSE: Preservation of the anal sphincter in surgery for cancer of the distal rectum in an attempt to avoid colostomy has been a main concern of colorectal surgeons. Various proposed procedures contradict oncological principles, especially with respect to pelvic lymphadenectomy. Therefore, prior knowledge of pelvic lymph node involvement is an important factor in choosing the operative technique, i.e., radical or conservative resection. Introduction of ultrasound, computerized tomography, and magnetic resonance have made preoperative study of the area possible. Nevertheless, these resources offer information of an anatomical nature only. Lymphoscintigraphy enables the morphological and functional evaluation of the pelvic area and contributes toward complementing the data obtained with the other imaging techniques. The objective of this prospective study is twofold: to standardize the lymphoscintigraphy technique and to use it to differentiate patients with rectal cancer from those with other coloproctologic diseases. CASUISTIC AND METHODS: Sixty patients with various coloproctologic diseases were studied prospectively. Ages ranged from 21 to 96 years (average, 51 and median, 55 years). Twenty-six patients were male and 34 were female. Thirty patients had carcinoma of the distal rectum as diagnosed by proctologic and anatomic-pathologic examinations, 20 patients had hemorrhoids, 5 had chagasic megacolon, 2 had diverticular disease, 2 had neoplasm of the right colon, and 1 had ulcerative colitis as diagnosed by proctologic exam and/or enema. The lymphoscintigraphy method consisted of injecting 0.25 mL of a dextran solution marked with radioactive technetium-99m into the right and left sides of the perianal region and obtaining images with a gamma camera. The results were analyzed statistically with a confidence level of 95% (P < .05) using the following statistical techniques: arithmetic and medium average, Fisher exact test, chi-square test corrected for continuity according to Yates, and distribution tables for the number of patients. RESULTS: In rectal cancer, the tracer progresses unilaterally or is absent; in other patients, the progress of the tracer is bilateral and symmetrical, although its progress may be slow. Statistical tests showed with high significance that the agreement index between the clinical diagnosis and the result of the lymphoscintigraphic exam was 93%. CONCLUSIONS: Lymphoscintigraphy is a standardized, painless, and harmless test that can be performed in all cases; it differentiates patients with rectal cancer from those with other coloproctological diseases.

Reconnaissance study of macrofossils from the upper purus river - Western Amazônia.

Fossils of wood, bone and teeth found along the Upper Purus River οf Amazonia. were studied using conventional microscopy and scanning electron microscopy. Mass spectometry was also used to investigate minor and trace element signatures of bone samples.The microsopy studies showed that there was little alteration of original textures. In the fossil wood samples, identified In thin section as tropical hardwood trees, the replacement of the original material with siderite suggests that fossilization occured in shallow sediments in which interstitial waters were saturated with respect to iron carbenate. In samples of both fossilized bone and wood, precipitation of secondary iron phases was commonly observed in cracks and voids. Other secondary phases Included silica, iron oxides, manganese carbonate. The intimate assοciation οf these secondary phases with the original biological structures could be evidence for a microbiological role in the formation of these phases. The similarity in rare earth element (REE) signatures for 2 fossil bone samples from different modern locations indicates their having shared similar diagenetic histories.The virtually complete preservation of original textures suggests that microscοpic studies could be useful in classifying fossil and even in identifying original materials. Rare carth signatures in fossilized bone may reflect ground water compositions at the time of fossilization.

Myxoma of the mitral valve

Only rarely do myxomas originate from the mitral valve. This is the report of a 49-year-old woman presenting with congestive heart failure. The diagnosis of an intracardiac tumor involving the anterior cuspid of the mitral valve was made by transesophageal echocardiography. The patient underwent surgery for tumor resection and plasty of the valve was made with reconstruction and preservation of the valve. The diagnosis of myxoma was confirmed by histology. This is the 23rd case of myxoma of the mitral valve reported in the literature.