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Amplification of DNA from soil is often inhibited by co-purified contaminants. A rapid, inexpensive, large-scale DNA extraction method involving minimal purification has been developed that is applicable to various soil types (1). DNA is also suitable for PCR amplification using various DNA targets. DNA was extracted from 100g of soil using direct lysis with glass beads and SDS followed by potassium acetate precipitation, polyethylene glycol precipitation, phenol extraction and isopropanol precipitation. This method was compared to other DNA extraction methods with regard to DNA purity and size.

The article describes a method for extracting DNA from soil which involves minimal purification prior to polymerase chain reaction (PCR) amplification. A fundamental obstacle to understanding microbial ecology and diversity is the inability to culture microorganisms from environmental samples. Extraction methods investigated include sonication, enzymatic lysis and extraction from bacterial cells isolated from soil. A comparison of extraction methods using a single soil collected from Macquarie University is presented.

c.m. davies, n. altavilla, m. krogh, c.m. ferguson, d.a. deere and n.j. ashbolt. 2004.To generate field-relevant inactivation rates forCryptosporidiumoocysts in soil that may serve as parameter values in models to predict the terrestrial fate and transport of oocysts in catchments.The inactivation ofCryptosporidiumoocysts in closed soil microcosms over time was monitored using fluorescencein situhybridization (FISH) as an estimate of oocyst‘viability’. Inactivation rates forCryptosporidiumin two soils were determined under a range of temperature, moisture and biotic status regimes. Temperature and soil type emerged as significantly influential factors (P < 0·05) forCryptosporidiuminactivation. In particular, temperatures as high as 35°C may result in enhanced inactivation.When modelling the fate ofCryptosporidiumoocysts in catchment soils, the use of inactivation rates that are appropriate for the specific catchment climate and soil types is essential. FISH was considered cost-effective and appropriate for determining oocyst inactivation rates in soil.Previous models for predicting the fate of pathogens in catchments have either made nonvalidated assumptions regarding inactivation ofCryptosporidiumin the terrestrial environment or have not considered it at all. Field-relevant inactivation data are presented, with significant implications for the management of catchments in warm temperate and tropical environments. [ABSTRACT FROM AUTHOR]

Background: Histology is still considered the golden standard for the diagnosis of H. pylori associated chronic gastritis and to evaluate the gastric mucosal damage. Serum pepsinogens I (sPGI)and II (sPGII), and gastric permeability investigated by oral sucrose test were found alterated in chronic superficial and deep gastritis as well of in presence of atrophy. Aim: To assess the integrity of gastric mucosa by two non-invasive tests: serum pepsinogen levels and oral sucrose test, and to investigate its relationship with histological findings. Patients and Methods: A series of 24 consecutive out patients (12 females, 12 males; mean age 42 years, range 18-81) observed at our department for upper gastrointestinal symptoms were enrolled. All patients were submitted to upper gastrointestinal endoscopy with biopsies examined by one patologist according to the Updated Sydney System classification, and performed oral sucrose test using 40 mg in 100 ml of water; serum samples were analysed using panel test including of EIA assays for sPGI and sPGII (Biohit, Helsinki, Finland). In first time, the patients were divided in Hp+ve and Hp -ve group and in second time in three groups:1. normal histology,2. superficial gastritis,3. focal or diffuse atrophic gastritis. Results: Fifteen patients resulted Hp +ve;a statically significant difference was found between sPGI, sPGII and sucrose in Hp+ve patients compared with Hp-ve ones (sPGI 147.7 ± 52.0 vs 80.0±36.4 p=0.0006; sPGII 16.8±9.0 vs 7.4±3.5 p=.007, sucrose 0.39±0.39 vs 0.07±0.006 p=0.007). A statically significant difference had been found for sucrose test singling out the patients according to different hystological severity of damage;normal histology (7 patients): sucrose 0.07±0.07; superficial gastritis (10 patients): 0.34±0.41 and atrophic gastritis (7 patients): 0.37±0.39; p=0.06. Conclusions: Serum pepsinogen I and II and sucrose test could be proposed as... [ABSTRACT FROM AUTHOR]

Background: Inflammation induced by Helicobacter pylori (Hp) increases gastric permeability and so molecule such as sucrose can pass the gastric epithelium. Permeability can be evaluated by oral sucrose test. Serum pepsinogen I and II levels have been described alterated in subjects with Helicobacter pylori related gastritis. Aim: to evaluate in Hp positive and Hp negative patients, serum pepsinogen I and II levels and gastric permeability, using sucrose test. Patients and Methods: fifty consecutive patients with dyspeptic symptoms and without antisecretory therapy (28 females, 22 males, mean age 39.2 years, range 17-81) were enrolled as outpatients in our department. A blood sample was collected and the panel (Biohit, Helsinki-Finland) by means of ELISA method was used for determination of pepsinogen I (sPGI), II (sPGII), and specific IgG serum levels; all patients were subsequently submitted to oral sucrose test: the fraction of the given oral dose of sucrose (40 gr) was evaluated in the urine. The Hp status as been assessed by means of histology, HpSA and/or specific IgG. Results: overall, no correlation was found between sPGI and sPGII levels and sucrose. Nineteen Hp positive patients showed this mean values: sPGI: 153 mg/dl (r =0.12), sPGII: 16,8 mg/dl (r =0.23) and sucrose: 0.126. In 31 Hp-ve patients mean values were: sPGI: 92,6 mg/dl (r =0.39), sPGII: 9.4 mg/dl (r =0.43) and sucrose: 0.126. Conclusions: these data confirmed the relationship between alterations of gastric mucosa induced by Helicobacter pylori and increased levels of pepsinogens and sucrose. No correlation was found between the two series, suggesting that possibly sPGI and sPGII were more related with infammation while sucrose could be alterated according with the severity of mucosal damage. [ABSTRACT FROM AUTHOR]

Potassium is a cation involved in the resting phase of membrane potential. Diets rich in fresh fruit and vegetables, whole grains, dairy products, and coffee have high potassium content. The shift from a pre-agriculture diet to today's consumption has led to reduced potassium intake. Indeed, the Western diet pattern is characterized by a high daily intake of saturated fats, sugars, sodium, proteins from red meat, and refined carbohydrates with a low potassium intake. These reductions are also mirrored by high sodium intakes and a high consumption of acid-generating food, which promote a chronic state of low-grade metabolic acidosis. The low-grade metabolic acidosis is a cause of the bone-wasting effect. Therefore, a long-standing acidotic state brings into play the bone that contributes to the buffering process through an increase in osteoclastic resorption. In consideration of this background, we carried out a review that focused on the pathophysiological mechanisms of the relationship between dietary potassium intake and bone health, underlining the detrimental effects of the Western dietary patterns characterized by low potassium consumption.

Hypertension (Htn) is a crucial cause of cardio-vascular and chronic kidney disease. Moreover, it is an independent risk factor for nephrolithiasis (NL). A diet rich in vegetables and fruits is indicated for both Htn and NL prevention, and the 24-h urinary potassium excretion can be used as a warning light for adherence. The aim of this study is to demonstrate the association between urinary potassium excretion and recurrent nephrolithiasis among patients affected by Htn. We have analyzed medical records of 119 patients affected by Htn and NL (SF-Hs) referring to Bone and Mineral Metabolism laboratory and 119 patients affected by Htn but without NL (nSF-Hs) referring to Hypertension and Organ Damage Hypertension related laboratory, both in Federico II University of Naples. The potassium 24-h urinary levels in SF-Hs were significantly lower compared to nSF-Hs. This difference was confirmed by the multivariable linear regression analysis in the unadjusted model and adjusted model for age, gender, metabolic syndrome, and body mass index. In conclusion, a higher potassium urinary excretion in 24-h is a protective factor against NL in patients affected by Htn and dietary interventions can be considered for kidney protection., (© 2023. The Author(s).)

The dispersion and initial transport of Cryptosporidium oocysts from fecal pats were investigated during artificial rainfall events on intact soil blocks (1,500 by 900 by 300 mm). Rainfall events of 55 mm h(-1) for 30 min and 25 mm h(-1) for 180 min were applied to soil plots with artificial fecal pats seeded with approximately 10(7) oocysts. The soil plots were divided in two, with one side devoid of vegetation and the other left with natural vegetation cover. Each combination of event intensity and duration, vegetation status, and degree of slope (5 degrees and 10 degrees ) was evaluated twice. Generally, a fivefold increase (P < 0.05) in runoff volume was generated on bare soil compared to vegetated soil, and significantly more infiltration, although highly variable, occurred through the vegetated soil blocks (P < 0.05). Runoff volume, event conditions (intensity and duration), vegetation status, degree of slope, and their interactions significantly affected the load of oocysts in the runoff. Surface runoff transported from 10(0.2) oocysts from vegetated loam soil (25-mm h(-1), 180-min event on 10 degrees slope) to up to 10(4.5) oocysts from unvegetated soil (55-mm h(-1), 30-min event on 10 degrees slope) over a 1-m distance. Surface soil samples downhill of the fecal pat contained significantly higher concentrations of oocysts on devegetated blocks than on vegetated blocks. Based on these results, there is a need to account for surface soil vegetation coverage as well as slope and rainfall runoff in future assessments of Cryptosporidium transport and when managing pathogen loads from stock grazing near streams within drinking water watersheds.

Lactoferrin is an iron binding protein involved in a large spectrum of biological actions including antimicrobial actions. Lactoferrin plays a central role in ferrokinetics: it binds free iron with great affinity limiting the amount of ions available for microorganism's metabolism. Its role in the host defence mechanisms consists in bacteriostatic and bactericidal effects; moreover it inhibits the proliferation of other microbes such as fungi and viruses. Lactoferrin is also involved in the modulation of immune system and recent studies indicate that lactoferrin directly modulates both production and function of neutrophils and monocytes.

Objectives: To clarify the relationship between interferon-alpha (IFN-alpha) therapy and autoimmune thyroiditis in chronic hepatitis C virus (HCV) infection, we investigated a selected number of patients without basal thyroid dysfunctions., Materials and Methods: 130 patients (average age: 20-70), with chronic HCV infection and without basal clinical and laboratory signs of autoimmune thyroiditis were divided into two groups: IFN-alpha treated (A) and untreated (B) patients. Group A received IFN-alpha (three million U.I./3 times a week) for six months; group B was followed for the same period. Thyroid peroxidase and thyroglobulin autoantibodies were measured by radioimmunoassay; thyroid function was measured by radioimmunoassay (free thyroxine and triiodothyronine) and immunoradiometric assay (thyroid stimulating hormone)., Results: After a 6-month period, thyroid autoantibodies positivity was documented in 21.1% of group A and in 10.3% of group B patients, both statistically relevant (p<0.001 and p<0.011, respectively). The comparison between the two groups was not statistically relevant (p=0.142)., Conclusions: Our study showed a prevalence of de novo thyroid autoimmunity in chronic HCV patients treated with IFN-alpha, confirming previous data in literature. The lack of a significant difference between treated and untreated patients strongly suggests that the anti-thyroid autoimmune response is linked to the HCV infection itself. Moreover, IFN-alpha therapy probably does not represent a risk factor in renewing the autoimmune processes of the thyroid gland. Thyroid function and autoantibodies must be systematically monitored in patients with HCV infection, especially in female and IFN-alpha treated population, not only to verify the possible thyroid abnormalities but also to rule out concomitant autoimmune diseases.

Background: Aim of the study was to assess the correlation between clinical stage of HCV-related liver disease and viraemia to immune response to different viral antigens., Methods: We considered 1330 patients with HCV chronic infection followed up from 6 months up to 6 years divided into two groups according to RIBA 3 (Abbott) response: Group I, 1231 patients with positivity for at least two bands (83 subjects with asymptomatic infection, 941 with chronic hepatitis, 201 with cirrhosis and 6 with HCC); Group II, 99 patients with positivity at only one band (45 with asymptomatic infection, 53 with chronic hepatitis and 1 cirrhotic)., Results: We noticed a major percentage of positive patients for at least three bands in more severe clinical forms (90% of chronic hepatitis or cirrhosis versus 60% of asymptomatics, p < 0.005, chi 2 test). Moreover we noticed a percentage increase of positivity for antibodies anti-c100 and anti-NS5 with the progression of liver damage, statistically significant differences between asymptomatics and patients with chronic forms. We also observed that viraemia is related neither to clinical stage nor to different reactivity to RIBA 3, albeit viraemia is usually detected more frequently among patients with liver damage, but unrelated to different reactivities., Conclusions: Our results show a clear correlation between number of reactivities towards HCV proteins and progression of liver damage, pointing out that immune response plays a direct role in the long-term outcome of HCV infection.

The ability of circulating mononuclear cells from recipients of HLA identical sibling marrow transplants to generate messenger (m) RNA for the haemopoietic growth factors interleukin-3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF) upon mitogen stimulation was investigated. The amount of IL-3 mRNA per ml of blood and IL-3 mRNA per 10(7) mononuclear cells as well as the amount of GM-CSF mRNA per ml of blood was significantly lower in transplant recipients than in normal volunteers. Lower values for mRNA expression for both IL-3 and GM-CSF were associated with the use of immunosuppressive therapy post transplant. No correlation was found between the expression of message for either cytokine, or the presence or absence of acute graft-versus-host disease at the time of testing. While there was no correlation between the quantity of GM-CSF message produced and time elapsed post transplant, IL-3 message expression increased slightly with increasing time post transplant. These defects of haemopoietic regulators constitute another parameter of impaired cellular immunity occurring after allogeneic marrow transplantation.