Implication of HLA-G 3' untranslated region polymorphisms in human papillomavirus infection.

Human papillomavirus (HPV) infection is involved in cervical lesion development. It interferes with host immune response and modifies the expression of human leukocyte antigen-G (HLA-G), a nonclassical HLA-I antigen with immune-inhibitory functions. We analyzed the frequencies of two HLA-G 3' untranslated region polymorphisms (14 bp ins/del, +3142C>G), involved in HLA-G modulation, in 33 condyloma acuminatum, 14 low grade squamous intraepithelial lesion and 100 invasive cervical cancer (ICC) HPV infected patients. We showed the involvement of HLA-G polymorphisms in HPV infection and lesion development, and suggested that 14 bp del allele promotes high-risk HPV infection, with del/C haplotype associated with ICC development. On the basis of these evidences, HLA-G polymorphisms could represent a risk factor in HPV positive subjects.

An accurate and reliable real time SNP genotyping assay for the HLA-G +3142 bp C>G polymorphism.

Human leukocyte antigen (HLA)-G is a non classical HLA class I antigen with immuno-modulatory functions. The HLA-G gene is characterized by a +3142C>G variant in the 3' untranslated region which is suggested to control protein production and to be associated with pathological conditions. DNAs form 221 randomly selected healthy subjects were genotyped for HLA-G +3142C>G polymorphism by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) (BaeGI), real-time PCR and sequencing. The 19% of the PCR-RFLP heterozygous samples were genotyped as 3142GG by real-time PCR and sequencing. This disagreement is caused by digestion efficiency in PCR-RFLP. This real-time PCR method will guarantee an accurate genotyping for future research and clinical purposes, where large cohorts should be tested.

Panbacterial real-time PCR to evaluate bacterial burden in chronic wounds treated with Cutimed™ Sorbact™.

The impact of polymicrobial bacterial infection on chronic wounds has been studied extensively, but standard bacteriological analysis is not always sensitive enough. Molecular approaches represent a promising alternative to the standard bacteriological analysis. This work aimed to assess the usefulness of a panbacterial quantitative real-time PCR reaction to quantitate the total bacterial load in chronic wounds treated with Cutimed™ Sorbact™, a novel therapeutic approach based on hydrophobic binding of bacteria to a membrane. The results obtained by panbacterial real-time PCR on conserved sequences of the bacterial 16S gene show that the bacterial burden significantly decreased in 10 out of 15 healing chronic wounds, and did not change in 5 out of 5 non-healing chronic wounds. On the contrary, classical culture for S. aureus and P. aeruginosa, and real-time PCR for Bacteroides and Fusobacterium did not show any correlation with the clinical outcome. Our study also shows that quantification of chronic wounds by panbacterial real-time PCR is to be performed on biopsies and not on swabs. These results show that panbacterial real-time PCR is a promising and quick method of determining the total bacterial load in chronic wounds, and suggest that it might be an important biomarker for the prognosis of chronic wounds under treatment.

Massive pulmonary embolism as a rare complication of a stab in the inguinal region in a HIV-positive patient: a case report.

OBJECTIVE: Venous thromboembolism (VTE) is a severe preventable disease; HIV-infection represents a prothrombotic condition, because of specific factors due to the virus itself, the host response and the antiretroviral therapy. Our aim is to raise awareness of thromboembolic risk when dealing with HIV-positive patients presenting to the Emergency Department for treatment of injuries, even though small. CASE REPORT: We present a case of a 33-year-old woman suffering from HIV-infection who presented to the Emergency Department with two small stab wounds. Laboratory tests and radiologic examinations were normal. About 8 hours after admission the patient developed a syncopal attack: a CT scan performed after hemodynamic stabilization revealed a massive pulmonary embolism (PE); the patient was then transferred to the Intensive Care Unit and treated with systemic thrombolysis. CONCLUSIONS: This case confirms that HIV-positive patients carry a higher risk for VTE and PE compared to general population, similarly to patients suffering from cancer: emergency physicians must be aware even in case of minor wounds.

Inhibition of human low-density lipoprotein oxidation by caffeic acid and other hydroxycinnamic acid derivatives.

The antioxidant activity of the major phenols derived from hydroxycinnamic acid (caffeic, ferulic, and p-coumaric acids) on in vitro LDL oxidation was screened, using Cu2+ as catalyst. The presence of the second phenolic hydroxy group enhanced the inhibitory effect of these compounds. In fact, at 5 microM concentration, only caffeic acid completely protected LDL from modification as measured as conjugated dienes formation and apo B-100 fragmentation, also preserving alpha-tocopherol. The effect of caffeic acid in inhibiting LDL oxidative modification induced by three different oxidant systems was tested. Using both Cu2+ and 2,2'-azobis (2-amidinopropane)-hydrochloride (AAPH), the inhibitory effect of caffeic acid was dose-dependent. Yet, the better protection was achieved in the metal-ion dependent system. Also the murine macrophages-mediated LDL oxidation was efficiently inhibited by 5 microM caffeic acid. UV-VIS spectra of caffeic acid incubated with cupric ions show the formation of a caffeic acid:copper complex, responsible for a transient chelating activity. This mechanism, coupled with its free radical scavenging property, accounts for the higher inhibitory activity exhibited by caffeic in Cu(2+)-catalyzed reaction.

Effect of caffeic acid on tert-butyl hydroperoxide-induced oxidative stress in U937.

Nonvitamin phenolic compounds are ubiquitous in food plants and therefore potentially present in human plasma in a diet-dependent concentration. The aim of this study was to evaluate the ability of caffeic acid, a phenolic acid with antioxidant activity, to affect cellular response in U937 human monocytic cells to t-butyl hydroperoxide-induced oxidative stress. In our experimental conditions caffeic acid was incorporated into cells without any cytotoxic effect. Caffeic acid-treated cells showed an increased resistance to oxidative challenge, as revealed by an higher percent of survival and the maintenance of an higher proliferative capacity in respect to control cells. This effect seems to be due to the ability of caffeic acid to reduce glutathione depletion and to inhibit lipid peroxidation during tBOOH treatment. It can be concluded that caffeic acid exerts an antioxidant action inside the cell, responsible for the observed modulation of the cellular response to oxidative challenge. Due to its presence in the diet, therefore, caffeic acid may play a role in the modulation of oxidative processes in vivo.

[Synchronous neoplasms of the colon and kidney: analysis of 2 case reports]. / Neoplasie sincrone del colon e del rene: analisi di due casi osservati.

Up until only a few decades ago synchronous renal cell carcinoma associated with primary tumors of other organs was diagnosed almost exclusively in necropsy series. Recently the widespread use of ultrasonography and CT has permitted diagnosis of clinically silent renal cell carcinoma in patients undergoing a work-up for other primary tumors. We report two cases of synchronous colon and renal cancer: 1) a 75 year old woman presented bilateral pulmonary nodules at chest X-Ray and stratigraphy. Total-body CT scan showed multiple, apparently metastatic, bilateral pulmonary lesions and two tumors in the right kidney and ascending colon. A right nephrectomy and right hemicolectomy were performed. She succumbed after 18 months as a result of metastatic spread to the liver, lungs and bone; 2) a 64 year old man was discovered to have a colon cancer during a barium enema examination and endoscopy. CT scan of the abdomen yielded an incidental diagnosis of a tumor in the left kidney. A left nephrectomy and left hemicolectomy were performed. After 6 months CT revealed no evidence of recurrence or metastases. Histology findings revealed two primary malignancies in both cases: a clear cell renal carcinoma and an adenocarcinoma of the colon.

[Use of mechanical staplers in the surgical treatment of rectal cancer in the elderly]. / L'utilizzo delle suturatrici meccaniche nel trattamento chirurgico del cancro del retto nell'anziano.

The aim of this study was to analyze the changes in surgical treatment of rectal cancer and in postsurgical complications as a result of the introduction of staplers in surgical practice, with particular reference to elderly patients (> or = 75 years). Since 1976, 320 patients have undergone rectal surgery in our department (207 colo-rectal or colo-anal anastomoses and 113 Miles operations have been performed). The patients were subdivided into two groups on the basis of age (> or = and < 75 years) and then further divided into subgroups operated on before and after 1983 (the year staplers were introduced). The morbidity rate (fistulas) was 16% in patients < 75 years old and 12% in elderly patients (P = n.s.); the mortality rate was 1% in patients < 75 years old and 12% in the more elderly group (P < 0.001). The utilization of staplers in surgical treatment of rectal cancer allows the surgeon to perform sphincter-sparing resections even in elderly patients without any changes in specific morbidity. Nevertheless, the complications are more severe in the elderly, with a higher mortality rate.

[Lymphadenectomy in the differentiated thyroid carcinoma]. / Linfectomia nei carcinomi differenziati della tiroide.

BACKGROUND: In this study the authors analyze the indications and the type of surgical procedure to perform on the neck lymph nodes in cases of differentiated thyroid carcinoma. METHODS: The study has been carried in a retrospective way. Between 1993 and 2001, 93 differentiated thyroid cancer were observed. There were 72 women and 21 men, with a mean age of 45.9 years (range 18-77). Patients have been divided into three groups: the first included 25 patients who underwent only thyroidectomy (24 total thyroidectomy, 1 lobectomy); the second group included 52 patients who underwent total thyroidectomy and lymphadenectomy of the central compartment. The third group included 16 patients who underwent total thyroidectomy and functional neck dissection. The median follow-up was 65.9 months. RESULTS: The postoperative complications was similar between three groups. Follow-up has shown similar survival and recurrences between groups. CONCLUSIONS: The elective lymphadenectomy of the central could be a solution for a routine treatment of differentiated thyroid cancer without lymph node involvement. The presence of cervico-lateral node metastases imposes a functional neck dissection.

Detection of HHV-6-specific mRNA and antigens in PBMCs of individuals with chromosomally integrated HHV-6 (ciHHV-6).

After inheritance of chromosomally integrated HHV-6 (ciHHV-6), viral DNA is found in every nucleated cell. The prevalence of ciHHV-6 is estimated to be 0.2-5% of humans. There are conflicting data on the potential for replication, possibly leading to clinical implications. We analysed peripheral blood mononuclear cells (PBMCs) from individuals with ciHHV-6 proven by fluorescence in situ hybridization (FISH) for HHV-6-specific mRNA (U94, U42, U22) and antigens by means of reverse transcription PCR and an indirect immunoperoxidase staining. U94 transcripts indicative of latent infection were detected in six (54.5%) out of 11 individuals at least once. Transcripts indicative of lytic infection (i.e. U42 and U22) were detected in four (36.4%) out of 11 individuals at least once. HHV-6 antigen was detected in seven (70%) out of 10 individuals at least once. The presence of viral mRNA and proteins supports virus gene expression from ciHHV-6, which may lead to virus replication. Considering the properties of active HHV-6 infection together with obvious replicative activity in individuals with ciHHV-6, pathophysiological effects leading to clinical consequences of chromosomally integrated viral DNA might be considered.

Effect of dietary oils on lipid peroxidation and on antioxidant parameters of rat plasma and lipoprotein fractions.

In order to investigate the influence of fatty acid pattern and antioxidants other than vitamin E on lipid peroxidation and antioxidant levels of plasma very low density and low density lipoproteins (VLDL + LDL), the effects of three diets (equalized for vitamin E) containing soybean oil, olive oil, or an oleate-rich mixture of triglycerides (triolein) were studied in rats. A significantly lower concentration of thiobarbituric acid-reactive substances (TBA-RS) in plasma and lipoproteins was found after the olive oil diet (soybean oil, 3.7 +/- 0.4 nmol/ml; triolein, 2.1 +/- 0.5 nmol/ml; olive oil, 1.5 +/- 0.3 nmol/ml, in plasma) (soybean oil, 0.99 +/- 0.16 nmol/ml; triolein, 0.96 +/- 0.13 nmol/ml; olive oil, 0.38 +/- 0.12 nmol/ml, in the VLDL + LDL fraction). Furthermore, the results from in vitro copper-induced lipid peroxidation, expressed in terms of conjugated dienes, lipid hydroperoxides, and TBA-RS content, showed that VLDL + LDL particles from olive olive oil-fed rats were remarkably resistant to oxidative modification. The results suggest that the fatty acid unsaturation of dietary oils is not the only determining factor of the antioxidant capacity of lipoproteins in this animal model. The maximal protection observed after the olive oil diet may be explained by the presence of other unidentified antioxidants in addition to vitamin E, derived from oil intake. Therefore, the optimal balance between the content of unsaturated fatty acids and natural antioxidants in dietary oils appears to be of major importance.

Effect of caffeic acid dietary supplementation on the antioxidant defense system in rat: an in vivo study.

Dietary supplementation of caffeic acid (0.2 and 0.8% w/w) in rats resulted in a statistically significant increase of alpha-tocopherol both in plasma and lipoprotein. While caffeic acid was not detectable in plasma under fasting conditions, in postprandial plasma it was present at micromole concentrations, doubling plasma total antioxidant capacity. Lipoproteins from caffeic acid-fed rats were more resistant than control to Cu2+-catalyzed oxidation, despite the lack of incorporation of caffeic acid in the particles. No significant effects on plasma and liver copper concentration, nor the increase in liver of Mn-superoxide dismutase reported in copper deficiency, were detected. These results demonstrate the physiological relevance of caffeic acid and its antioxidant action in vivo, through both a direct contribution to the antioxidant defense system and a sparing effect on alpha-tocopherol.

Evaluation of protein quality: comparison of single-point and multi-point assays.

The quality of different protein sources having a wide spectrum of potential biological values has been assessed with multi-point and singly-point assays on growing Sprague-Dawley rats weighing 60 g. Values obtained with multi-point assays, expressed goth as absolute and relative values, were well correlated irrespective of the response parameter, i.e. as absolute and relative values, were well correlated irrespective of the response parameter, i.e. body weight change, body water or body nitrogen (r = 0.980). Single-point assay values based on body nitrogen content were also well correlated with the multi-point assays, but some discrepancies were noted for what assays based on body weight change was concerned. The modification of body protein concentration, in particular of rats fed the protein-free diet, was the main cause of these discrepancies. The problem of reference protein, when protein quality was expressed as relative value, was also discussed.

Influence of dietary thermally oxidized soybean oil on the oxidative status of rats of different ages.

Young and aging rats were fed for different periods (10, 90, 180 and 365 days) diets containing 15% of fresh or heated soybean oil. Thiobarbituric reactive substances (TBA-RS), lipofuscin, superoxide dismutase (SOD), vitamin A, vitamin E and microsomal and mitochondrial fatty acids in liver, brain and serum were measured. Heated oil diets induced significant increase of TBA-RS levels in liver, with earlier effects in aging rats and affected SOD activity in aging rats only after a long period of feeding. Circulating and stored vitamin A were reduced in both young and aging rats, with earlier effects in young animals. Serum and liver vitamin E was significantly reduced in all test groups. The results indicate that heated unsaturated oil produces reduction in the antioxidative defense system and that vitamin E status is the earliest indicator of the oxidative effect regardless of age.

Effect of fish oil and coconut oil on antioxidant defence system and lipid peroxidation in rat liver.

Diets high in fish oil containing polyunsaturated fatty acids of the n-3 family, have been suggested to decrease the risk of cardiovascular disease. However these lipids are highly susceptible to oxidative deterioration. In order to investigate the influence of n-3 fatty acids on oxidative status, the effect of feeding rats with fish oil or coconut oil diets was studied by measuring different parameters related to an oxidative free radical challenge. Synthetic diets containing 15% (w/v) fish oil or coconut oil were used to feed growing rats for 4 weeks. As compared to control diet, the fish oil containing diet produced a significant decrease of cholesterol and triglyceride concentration in serum, however there was a significant increase in lipid peroxidation products. In addition, in fish oil fed animals, there was also a decrease in vitamin E and A concentration. Furthermore, the rate of lipid peroxidation in isolated microsomes was three fold higher in rats fed fish oil as compared to rats with coconut oil diet. No significant differences between the two experimental groups were observed in superoxide dismutase (SOD) and phospholipid hydroperoxide glutathione peroxidase (PHGPX) activities. However, there was a decrease in glutathione peroxidase (GPX) activity. These results suggest that fish oil feeding at an amount compatible with human diet, although decreasing plasma lipids, actually challenge the antioxidant defence system, thus increasing the susceptibility of tissues to free radical oxidative damage.