[Topical treatment of vaginal infections by the association of metronidazole-clotrimazole]. / L'attualità del trattamento topico delle infezioni vaginali con l'associazione metronidazolo-clotrimazolo.

Vaginal infections are one of the most gynecological frequently diseases observed and with significant psychological and clinical implications. Their pharmacological treatment may require different options, but even today, scientific literature and international guidelines recommend the use of metronidazole for the treatment of bacterial vaginosis (BV) and trichomoniasis, and the clotrimazole for fungal infections from Candida (VVC). In this contest, the topical association of clotrimazole-metronidazole (vaginal pessaries, cream and douches) represents a current reference treatment for these types of infections with a number of important pharmacological properties. This combination allows an effective activity against to a broad spectrum of pathogens (bacterial, fungal and protozoan), a feature particularly relevant in the case of mixed infections. Furthermore it allows a synergistic action that improve the therapeutic abilities of the individual components, a reduction of the spontaneous resistance of some microorganisms and the activity against symptoms and signs of vaginal inflammation with maintaining the vaginal ecosystem, since they have no activity against endogenous lactobacilli. Finally, recent studies have shown the ability of the topical association of metronidazole-clotrimazole to inhibit the in vitro phenotypic switching of Candida albicans, and its effectiveness against Recurrent Vulvovaginal Candidiasis (RVVC).

The zinc finger protein Gfi-1 can enhance STAT3 signaling by interacting with the STAT3 inhibitor PIAS3.

STAT factors act as signal transducers of cytokine receptors and transcriptionally activate specific target genes. The recently discovered protein PIAS3 binds directly to STAT3 and blocks transcriptional activation. Here, we present experimental evidence implementing the zinc finger protein Gfi-1 as a new regulatory factor in STAT3-mediated signal transduction. The interaction between the two proteins first became evident in a yeast two-hybrid screen but was also seen in coprecipitation experiments from eukaryotic cells. Moreover, we found that both Gfi-1 and PIAS3 colocalize in a characteristic nuclear dot structure. While PIAS3 exerts a profound inhibitory effect on STAT3-mediated transcription of target promoters, Gfi-1 can overcome the PIAS3 block and significantly enhances STAT3-mediated transcriptional activation. In primary T cells, Gfi-1 was able to amplify IL-6-dependent T-cell activation. As Gfi-1 is a known, dominant proto-oncogene, our findings bear particular importance for the recently described ability of STAT3 to transform cells malignantly and offer an explanation of the oncogenic potential of Gfi-1 in T lymphocytes.

Biohumoral parameters and bone mineral content in the identification of high risk subjects for postmenopausal osteoporosis.

BACKGROUND: The aim of this study was to evaluate the possibility of identifying women with a high risk of postmenopausal osteoporosis by using computerised bone mineral analysis (CBMA) associated with markers of bone turnover in order to provide a valid and reliable screening test. METHODS: A total of 925 patients were evaluated, 252 of whom had already undergone a preliminary densitometric test six months earlier and were diagnosed as fast bone losers. 225 of them (89%) also showed altered bone turnover markers. CBMA was negative in the remaining 673 patients but 13 patients showed altered bone turnover markers and three of the latter then showed a positive CBMA 18 months later. The 673 patients who, after six months of study, were not fast bone losers were monitored over time. RESULTS: It emerges from these results that fast bone losers are characterised by higher levels of hydroxyprolinuria and calcium, lower levels of oestrone and estradiol, and reduced body weight compared to healthy subjects. CONCLUSIONS: This simplified method enabled 79% of the fast bone losers to be identified, whereas densitometry identified 87.5% of the high-risk subjects. The main advantage of our simplified method compared to the measurement of bone mineral content is that it identifies the majority of fast bone losers in the initial postmenopausal period, before a substantial reduction in BMC has taken place.

Mutational-screening in the factor VIII gene resulting in the identification of three novel mutations, one of which is a donor splice mutation. Mutations in brief no. 245. Online.

Hemophilia A is an X-linked bleeding disease caused by mutations in the coagulation factor VIII gene. The identification and characterization of pathogenic mutations allows the recognition of new mechanisms of functional disturbances of factor VIII. To screen for mutations exons 1-26 of the factor VIII gene have been amplified genomically and analyzed by SSCP followed by direct sequencing of respective exons showing abnormal electrophoretic mobility on SSCP analysis. In the present study we report the detection of four mutations in the factor VIII gene, of which three are novel. The mutational analysis of a patient with severe hemophilia A has revealed that the ac transversion at position 3 of the donor-splice-site of intron 23 results in the skipping of exon 23. A novel nonsense mutation Q1778X in exon 16 of factor VIII gene has been identified in a second hemophilia A case. Furthermore two missense mutations have been ascertained: a novel, S183R, causing a mild phenotype of hemophilia A and R282H, previously described in association with severe hemophilia A.

A deletion/insertion leading to the generation of a direct repeat as a result of slipped mispairing and intragenic recombination in the factor VIII gene.

A deletion/insertion in the human factor VIII gene was found in a patient with severe hemophilia A; 316 bp were removed, viz., those enclosing part of intron 15 and the first 7 bp of exon 16. In addition to the deletion, 6 bp were added to the deletion breakpoints; this resulted in the duplication of an existing 13-bp unit. Thus, an overlapping 13-bp direct repeat was generated at the deletion junction. Moreover, the deleted fragment itself was flanked by two homologous 6-bp sequences, one unit being lost by the deletion. A combination of slipped mispairing during replication and an intragenic recombination is discussed to describe this deletion/insertion process.

Molecular diagnostics of 15 hemophilia A patients: characterization of eight novel mutations in the factor VIII gene, two of which result in exon skipping.

The X-linked bleeding disorder hemophilia A is caused by mutations in the coagulation factor VIII gene. A high frequency of de novo mutations and the large size of this gene complicate the molecular diagnostic of hemophilia A. Characterization of mutations, however, may help identify amino acids or regions with essential functional or structural properties and thereby clarify the mechanism of pathogenesis. In the present study, we describe the identification of 15 mutations in the factor VIII gene, of which eight are novel. Among the patients with severe hemophilia A, two splice mutations (IVS5-3 and IVS19-2), a 4-bp deletion ((TACA) at codon 1215, and a missense mutation G1850V have been characterized. The missense mutations G479R, R531C, V537D, N2129S and I2190N were found for five patients with a moderate course of hemophilia A disease. A silent mutation resulting in activation of a cryptic acceptor splice site within exon 11 and four other missense mutations Y114C, R1689H, R2150H (2x), M2164V have been identified for six patients with mild hemophilia A.

Ultrasound diagnostic criteria in breast disease.

The Kasumi-Kamio parameters (margins, peripheral echoes, internal echoes, posterior echoes, lateral shadow cones) and the ratio between the longitudinal and transverse diameter of the breast lump form the basis for a list of standardised diagnostic criteria on which to base an analysis of breast disease that assesses the specificity and sensitivity of ultrasonography as a valid, reliable initial step in the diagnosis of breast tumours. The study was based on a series of 129 tumour cases and produced correct diagnosis in 89.28% of benign, 83.33% of malignant cases.

Mutational analysis of ectopic factor VIII transcripts from hemophilia A patients: identification of cryptic splice site, exon skipping and novel point mutations.

Mutational analysis of the gene for clotting factor VIII is complicated by its large size, the high frequency of de novo mutations and its tissue-specific expression. In order to facilitate the search for mutations, we have used a combination of reverse transcription-polymerase chain reaction (RT-PCR) of ectopic factor VIII transcripts, PCR of genomic DNA, single-strand conformation polymorphism analysis and direct sequencing. Here we describe the characterization of seven potentially pathogenic mutations: five of them are novel and the reason for the pathogenicity of the sixth could be determined. Here cDNA analysis revealed the absence of the first 47 bp of exon 16 in approximately 80% of the processed factor VIII mRNA, likely due to activation of a cryptic acceptor splice site within exon 16. The other novel mutations reported here include the skipping of exon 19, which predicts the removal of the corresponding 39 amino acids from the A3 domain, and four missense mutations: W14G, Y620C, W1889L, and Q2087R.

Alternative splicing in PAX2 generates a new reading frame and an extended conserved coding region at the carboxy terminus.

PAX2 is a member of the PAX multigene family encoding transcription factors active in specific tissues during embryogenesis. Several PAX/Pax genes (PAX and Pax describe homologous genes in human and mice, respectively) have been shown to possess critical morphogenetic functions as identified by the analysis of mice targeted for Pax genes and the phenotype of patients heterozygous for PAX mutations. Mutations in PAX2 have been shown to be implicated in independent cases of renal-coloboma syndrome. Here, we report the characterisation of a new PAX2 isoform, viz. PAX2d, which arises because of the use of an alternative acceptor splice site within exon 12 of the PAX2 gene; this leads to a shift in the reading frame. A conserved coding region extended over the regular stop codon may emphasize the biological significance of this isoform.

Endoscopic and histologic findings in the upper gastrointestinal tract of children with coeliac disease.

Frequency of mucosal damage to the esophagus, stomach, and duodenum was investigated in 176 children with coeliac disease (CD) during 230 upper GI endoscopies performed to obtain duodenal biopsy specimens and was compared with findings in 230 age-matched children who underwent endoscopy for upper GI complaints without CD (non-CD patients). To evaluate a possible association with gluten ingestion, we then compared frequency of mucosal damage in patients on a gluten-containing diet and those on a gluten-free Diet (GFD). In children with CD, frequency of esophageal damage seen at endoscopy and of peptic esophagitis shown by histology were significantly lower than in non-CD patients (p < 0.01) due to the very low frequency of mucosal damage in CD children on GFD; however, frequency of columnar metaplasia was significantly higher (p < 0.05). At endoscopy, CD children had a significantly lower frequency of gastric abnormalities, but histology showed a higher prevalence of superficial chronic gastritis (SCG; p < 0.01). SCG was associated with gluten ingestion, since its frequency in CD children on GFD was similar to the frequency in non-CD patients. At endoscopy, frequency of duodenal mucosal damage was similar in CD and non-CD patients. In addition to villous atrophy, histology showed a significantly higher frequency of duodenitis in CD children on a gluten-containing diet (p < 0.001 vs. non-CD patients; p < 0.05 vs. CD children on GFD). Our findings show that the mucosa of the whole upper GI tract can be damaged in CD patients and that the prevalence of some changes is higher with a gluten-containing diet.

T cell receptor heterogeneity in gamma delta T cell clones from intestinal biopsies of patients with celiac disease.

In celiac disease large numbers of gamma delta T lymphocytes infiltrate the intestinal epithelia. We have isolated intestinal gamma delta T cell clones from patients with celiac disease and have analyzed their T cell receptor repertoire. T cell lines and clones were obtained from jejunal biopsies of 14 celiac patients and 12 individuals without celiac disease. These were analyzed by staining with monoclonal antibodies against CD3, alpha beta and gamma delta T cell receptor, by Southern blot with gamma- and delta-specific probes and by polymerase chain reaction using V delta-specific oligonucleotides. Intestinal gamma delta cells from patients with celiac disease differed from those of controls with normal jejunal histology in that V delta 1+ cells and V delta 1-V delta 2- cells were significantly increased. There was no evidence of the expansion of one or more clones expressing particular types of gamma delta T cell receptor.

Effect of immunostimulating therapy on the immunocompetent system in breast carcinoma.

The authors studied 68 patients suffering from breast cancer, with or without lymph node metastasis, who underwent surgery and antitumour therapy (CMF). Twenty-three patients were treated using CMF and 1.5 mg/kg of thymostimulin, 24 with CMF and 1 mg/kg of thymostimulin and lastly, 21 subjects received anti-tumour therapy with CMF alone. Thymostimulin was administered every day for a week prior to surgery; subsequently, it was administered on alternate days for a week and then twice a week for 3 months. The blastogenesis of immunocompetent cells was evaluated. During thymostimulin treatment a higher rate of 3HTdR captation (p < 0.005) by cells stimulated with ConA + IL-2 was observed; these levels tended to increase after 3 weeks and reached statistically significant levels after 3 months of treatment; no significant changes were observed in those patients treated with CMF alone. In addition, the cytotoxic activity of monocytes and NK cells against K-562 cells and against short-lasting cell lines derived from breast carcinoma was also studied. It was observed that this activity increased significantly (p < 0.002) following thymostimulin treatment; this increase was greater in subjects treated with 1.5 mg/kg compared to those treated with 1 mg/kg, but the difference was not statistically significant. The study also evaluated the presence of IL-2 receptors (Tac): thymostimulin treatment for 3 months led to the appearance of receptors, although in restricted numbers, on non-stimulated cells. After IL-2 stimulation, the percentage of cells with Tac receptors increased significantly (p < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

[Adherence to a diet and the social aspects of patients with celiac disease]. / Aderenza alla dieta ed aspetti sociali dei pazienti con malattia celiaca.

We studied the social behaviour and dietary habits of 335 coeliacs older than 6 yrs diagnosed in our paediatric gastroenterology unit by a mailed questionnaire, 156 patients (45.2%) answered all questions; their median age was 14.7 yrs (range 6-29). We found that the disease does not compromise educational achievement and working capacity of patients. A majority of our coeliacs are students (from primary school to university) and rather successful ones since 55% of them passed their previous year examinations. Some are already employed and work as clerks, artisans, masons or skilled workers. 89.6% of our patients reported to be on a strict gluten-free diet, 9% introduce small amounts of gluten and 1.4% are on a normal diet by their own decision. Coeliac patients originating from Northern Italy have more of their gluten-free foods home made and use more gliadin free cereals (rice, maize), whereas coeliacs originating from the Southern regions consume more ready made gluten-free foods. We have assessed the amount of gluten-free products consumed monthly by our patients and their food preferences. Females eat less than males and prefer bread and flour based dishes, whereas males east more pasta and biscuits.

[Celiac disease in Piedmont. An epidemiological-clinical study]. / La malattia celiaca in Piemonte. Studio epidemiologico-clinico.

The incidence of coeliac disease (CD) was calculated on 304 patients under eighteen who were born in the city of Turin and its province in the years 1975-1989; the prevalence on 494 patients who live in the Piedmont Region. The mean crude yearly incidence was 0.511/1000 (1:1957 live birth). It varied from year to year, reaching minimum values in the years 1984-1987. In a contemporary epidemiological study, the mean crude incidence of CD in Italy was 1.2/1000 (1:833 live birth) twice the rate observed in Turin. The prevalence of paediatric CD in Piedmont was 113 per million inhabitants. Since CD has a normal life expectancy, its prevalence may be expected to increase. In the provinces of Novara, Alessandria and Asti CD prevalence was lower than in the others. Mean age at onset was 6 mos in 1975 and increased to 34 mos in 1989. Mean age at diagnosis was 15 mos in 1981, and 7 yrs in 1989. Symptoms were more numerous and severe in patients under 12 mos of age, and became fewer and often atypical in older children. We can therefore speculate that the trend towards a decreasing incidence of CD in recent years my be due to delayed diagnosis.

[Clinical data on celiac disease with an early or late onset]. / Dati clinici nella malattia celiaca ad esordio precoce o tardivo.

Typical symptoms in celiac disease (CD) are usually associated with early onset of the disease, whereas an atypical symptomatology has more often a later onset. The aim of this study was to evaluate the prevalence of some clinical signs and symptoms in children whose CD started before one year of age ("early onset" 135 children, M/F 50/85, mean age at onset 6.9 +/- 1.9 months) and in children whose disease started later ("late onset", M/F 14/26, mean age at onset 26.3 +/- 26.7 months). We analyzed: a) time lapse between gluten introduction and onset of symptoms, b) prevalence of patients with gastrointestinal symptoms alone and that of patients with gastrointestinal plus extraintestinal symptoms, c) frequency of each symptom. We then evaluated the influence of breast feeding and age of gluten introduction on time lapse. Our results showed that typical gastrointestinal symptoms, like diarrhea anorexia and abdominal distension prevailed both in children with early and late onset; whereas failure to thrive was significantly more frequent in children with an early onset CD (p < 0.01). Breast feeding delayed onset of symptoms: time lapse was significantly longer in children breast fed for a longer time (p < 0.001). On the contrary, age at first gluten ingestion seemed to have no influence on age at onset, since it was similar in both groups.

Treatment of childhood peptic oesophagitis with famotidine or alginate-antacid.

Both antacids and H2 receptor antagonists have a potential role in treating peptic oesophagitis associated with acid reflux. The aim of this study is to assess the efficacy and tolerability of famotidine and alginate-antacid in children with endoscopically documented peptic oesophagitis. We compared clinical, endoscopic and histological response to alginate-antacid (1 tablet to be chewed 30 min after each meal and at bed time) or famotidine (1 mg/kg before supper at 7 pm) in 49 children (mean age 9 years, 34 males). Twenty-four of them were randomly allocated to alginate-antacid treatment (group A), twenty-five to famotidine (group B). Both treatments were protracted for six months and endoscopy was then repeated. Symptoms disappeared in 43.4% and improved in 47.8% of children of group A and disappeared in 91.6% of those of group B (p less than .05). Mean time for symptom disappearance was 17 weeks for group A and 5 weeks for group B (p less than .01). After six months, at endoscopy oesophagitis was healed in 43.4% in group A and in 41.6% of group B, the difference between the two groups was not significant, however the improvement of endoscopic grades induced by famotidine was significantly better (p less than .05). Histology showed a healed peptic oesophagitis in 52.2% of the children in group A and in 70.8% of group B (p less than .001). No toxicity was observed with either treatment. We conclude that famotidine is superior to alginate-antacid in treating peptic oesophagitis associated with acid reflux, since it induces a better symptomatic response and a greater improvement of endoscopic lesions.

[Comparative analysis of therapeutic effects of carbocalcitonin and and conjugated estrogens in post-menopausal osteoporosis]. / Analisi comparativa degli effetti terapeutici della carbocalcitonina e degli estrogeni coniugati nell'osteoporosi post-menopausale.

In order to assess the correlation between menopause and osteoporosis, both in pathogenetic and therapeutical terms, a study was carried out in four comparable group of patients at Department B of the Institute of Gynaecology and Obstetrics at the University of Turin. Patients were divided as follows: 24 patents affected by evident osteoporosis, 39 patients with the first symptoms of osteoporosis, 27 with hypercalcemia and 33 healthy controls. The following tests were performed in all subjects: serum assay of androstenedione, estrone, 17-beta-estradiol, PTH, calcium, phosphorus, alkaline phosphatase and creatinine. Laboratory tests were repeated monthly in all patients and control subjects. Dual chromatic ray bone densitometry was performed in all patients at the start and end of treatment. With regard to therapy, each group was subdivided into two equal subgroups which were treated with carbocalcitonin or conjugated estrogens. From the findings, it is clear that there is a non-significant difference between serum levels of androstenedione, estrone and estradiol in the three groups examined and control subjects. Although the possibility that the fall in steroid hormones might contribute to bone load cannot be excluded, it is not possible to demonstrate that this is the most important factor in the pathogenesis of osteoporosis given that many women do not develop osteoporotic symptoms after menopause. In addition, in therapeutic terms, all bone density parameters considered in patient osteoporosis improved after therapy.(ABSTRACT TRUNCATED AT 250 WORDS)

Treatment of pelvic inflammatory disease with combined piperacillin and minocycline therapy.

Pelvic inflammatory disease (PID) has its aetiopathogenesis especially in anaerobic microorganisms and it is considered that 15% of patients with PID do not respond to initial treatment, 20% have at least one recurrence and 18% become sterile. Forty-six patients aged between 18 and 45 treated as out-patients with the association piperacillin-minocycline have been considered. These patients were monitored up to 5 weeks from the start of therapy following standard evaluation criteria and it is held that this therapy is effective and indicated as a first approach.

[Correlation between use of IUD and pelvic inflammatory disease (PID) as independent variables and risk of occurrence of ectopic pregnancy]. / Correlazione tra uso di IUD e pid quali variabili indipendenti e rischio di insorgenza di gravidanza ectopica.

PIP: The relationship between IUD and ectopic pregnancy was evaluated in a study involving 189 controls and 69 women with ectopic pregnancy. The patients involved were between the ages of 18 and 45, sexually active, without previous tubal ligation; they were interviewed about previous pregnancies, diseases, abortions and their socio-economic status. Diagnosis of ectopic pregnancy was made using serum pregnancy tests, needle biopsies of the Douglas pouch and/or ultrasound and/or laparoscope. The results show that there is no statistical difference between patients using the IUD with a history of previous PID and those with previous PID who had never used an IUD; instead, there is a 10-fold risk in women affected by PID, regardless if IUD use. Women who had used an IUD for less than 3 years showed a smaller risk than those who had used the IUD for a greater period of time. Finally there was no difference in women not affected by PID, regardless of IUD use. This ultimately correlates with the notion that one of the mechanisms of action of the IUD consists of an endometrial inflammation as seen in foreign-body reactions.^ieng