Influence of a preschool preventive dental programme on caries prevalence, oral care and secretory immunity to Streptococcus mutans in young adults.

OBJECTIVE: To evaluate oral hygiene habits, decayed, missing and filled teeth (DMFT) and surfaces (DMFS), dental care, dietetic habits and anti-Streptococcus mutans salivary secretory Immunoglobulin A (SIgA) in young adults who attended a preventive programme during preschool age. MATERIAL AND METHODS: The study group (Baby Clinic) comprised 72 patients, aged 18-25 years, who had participated in the Baby Clinic preventive programme. The control group was age- and gender-matched. The patients were examined and unstimulated whole saliva was sampled for detection of anti-S. mutansSIgA antibodies. RESULTS: Control patients presented increased DMFS scores (P < .05). Hygiene habits, cariogenic diet and antibody levels were not different between groups (P > .05). Baby Clinic patients presented better periodontal status (P < .005), less calculus (P < .005) and bleeding on probing (P < .005), and reported visiting dental services more regularly (P < .05). Adjusted multivariate linear regression analysis demonstrated that DMFT was associated with study group (P < .05), gender (P < .05), parents' education (P < .05), carbohydrate intake (P < .001) and levels of anti-S. mutansSIgA (P < .007). DMFS was associated with time elapsed since the last visit to the dentist (P < .005) and weekly carbohydrate intake (P < .005). CONCLUSION: Preventive programmes for preschool children positively impact on DMFS and periodontal status in young adults, but have no long-term effects on dietary or hygiene habits.

Atrioventricular block in acute inferior wall myocardial infarction: harbinger of associated obstruction of the left anterior descending coronary artery.

In a prospective study 51 consecutive patients who survived the acute phase of inferior wall myocardial infarction underwent coronary arteriography. Eleven patients developed some degree of atrioventricular (AV) block in the acute phase of infarction that disappeared within a few days and was considered by electrocardiographic analysis to be located in the AV node. Patients with AV block during acute myocardial infarction had a significantly higher prevalence of left anterior descending coronary artery obstruction (91 versus 55%, p less than 0.05) than did patients without AV block and the obstruction preceded the exit of the first septal perforator branch in 73% of cases with heart block and in 30% of cases without block (p less than 0.01). The sensitivity, specificity and predictive values were 31, 95 and 91%, respectively, for the existence of left anterior descending coronary artery obstruction when AV block occurred during acute inferior myocardial infarction, and 40, 90 and 73%, respectively, for the occurrence of the coronary artery obstruction before the exit of the first septal perforator branch. Patients with inferior myocardial infarction and left anterior descending coronary artery obstruction have a sixfold greater chance of developing heart block in the acute phase of infarction than do patients with inferior infarction without such obstruction (p less than 0.05). These findings also support the observations that the proximal AV conduction system usually has a dual arterial blood supply from both the right and left anterior descending coronary arteries, and may explain the transient behavior of heart block and lack of necrosis of the AV node seen in these patients.

Novel splicing and missense mutations in autosomal dominant polycystic kidney disease 1 (PKD1) gene: expression of mutated genes.

Autosomal dominant polycystic kidney disease (ADPKD) is a common disorder mostly characterized by cyst formation in kidney tubules. The majority of ADPKD cases is caused by mutations in the PKD1 gene, but no prevalent mutation has been reported. By heteroduplex analysis of the 3' single-copy region of the gene, we have searched for mutations in subjects from 40 ADPKD families of Northern Italy. Seven novel polymorphisms and three novel disease-associated mutations (R3718Q, L3851P and IVS45+56del25) were identified. Both missense mutations are located in the major extracellular loop of polycystin-1. The 25 bp deletion inside intron 45 did not affect 5' and 3' consensus splicing sites, but caused a 56 nucleotide out of frame-deletion due to activation of a cryptic 3' splice site in exon 46. The mutated RNA should produce a truncated polycystin 1 at the G binding peptide in the intracellular C-terminal end of the protein. RT-PCR analysis showed that the disease-associated mutations were present in transcribed sequences. In particular, RNA analysis of BHK cells transfected with PKD1 genomic DNA, including the deleted intron, showed that no normal transcript is produced by the deleted gene. This intronic mutation, found in a large pedigree, seems to be associated with a prevalence of cerebrovascular disease.

Isolate antibody to hepatitis C virus core antigen (C22) by RIBA-2: correlation with HCV-RNA and anti-NS5.

The presence of circulating hepatitis C virus genome (HCV-RNA), elevated ALT levels and antibodies to an NS5-derived synthetic peptide have been examined in 13 subjects with isolate positivity for antibodies to the HCV core antigen (C22) on RIBA-2 testing. All subjects were followed up for 8-18 months (mean 12.4 months). In seven subjects (54%), intermittent or persistent viremia was associated with abnormal ALT levels (6 subjects) and with positivity for antibodies to NS5-peptide (6 subjects). On the other hand, in 6 out of 13 subjects (46%) no viral replication, no liver cytonecrosis and no antibodies to NS5 were found. It is concluded that isolate reactivity to C22 by RIBA-2 is a heterogeneous condition that corresponds to two distinct categories of subjects: those with active HCV infection and those without evidence of virus replication. Although HCV-RNA determination is the most reliable means of identifying HCV carriers, antibodies to NS5 can be a useful marker of virus activity. In fact, antibodies to NS5 were detected in 6 out of 7 viremic patients, compared to 0 out of 6 non-viremic patients (P = 0.004). It remains to be elucidated whether the isolate reactivity to core antigen found in non-viremic subjects represents a specific, HCV-induced antibody response, or is an unrelated crossreactivity.

Distal extremity swelling with pitting edema in psoriatic arthritis: a case-control study.

OBJECTIVE: To evaluate the frequency and the clinical characteristics of distal extremity swelling with pitting edema in patients with psoriatic arthritis (PsA). METHODS: This was a case-control study of consecutive outpatients with PsA (old and new diagnosis) observed over a 3-month period in three secondary referral centers in Italy. As controls we used the two consecutive rheumatic outpatients, excluding those with spondylarthropathies, observed after a PsA patient. The demographic and clinical features were assessed by clinical examination and review of the medical records. RESULTS: A total of 183 patients with PsA and 366 controls were evaluated. Distal extremity swelling with pitting edema was recorded in 39/183 (21%) PsA patients and in 18/366 (4.9%) controls (p < 0.0001). In 8/39 (20%) patients this feature presented as a first, isolated manifestation of PsA, and in 8 others it was associated with other features of PsA at diagnosis. The upper and lower extremities were affected, predominantly asymmetrically, in 40% and 60% of the cases respectively. In patients with pitting edema compared to those without this feature, the frequency of Achilles enthesitis and plantar fasciitis, calculated together, was higher (p < 0.05) and the duration of arthritis was significantly lower (p = 0.02). In 7 patients the clinical evidence of a predominant involvement of tenosynovial structures was confirmed by MRI. CONCLUSION: Upper or lower distal extremity swelling with pitting edema due to tenosynovitis, usually unilateral, is a common feature in PsA patients and may represent the first, isolated manifestation of the disease.

CT identification of coeliac ganglia.

The authors achieved the "in vivo" identification of the coeliac ganglia (C.G.), using computerised tomography (CT). This result was confirmed by autopsies and by CT scans of an anatomical specimen in which the coeliac ganglia had been previously marked. CT allows an exact location of the coeliac ganglia and can be very useful for a precise alcoholic neurolysis of the coeliac plexus.

[Myocardial ischemia with stable angina pectoris: clinico-ergometric evaluation after the use of diltiazem]. / Isquemia miocárdica com angina do peito estável: avaliação clínico-ergométrica após uso de diltiazem.

PURPOSE: To evaluate the efficacy of diltiazem versus placebo in patients with stable angina. METHODS: Eight-seven angina pectoris patients, mean age of 57 +/- 9, 82 white and 79 male were evaluated in a randomized, double-blind trial of two groups of patients diltiazem and placebo, 3 to 4 tablets a day (diltiazem 180 to 240 mg daily). The patients were evaluated after laboratory tests and clinical-ergometric examinations. A coronary arteriography was performed on study entry. RESULTS: The average of anginal attacks, number of weekly sublingual nitrate, heart rate, systolic and diastolic pressure at rest and at the end of diltiazem period were significantly lower (p < 0.05) regarding same periods on placebo. The percentage of depression for ST-segment was lower for diltiazem when compared with placebo (p < 0.05) and the percentage of patients that reach higher stages in the ergometric test was significantly better for diltiazem. Heart rate and systolic plus diastolic pressures after exercise did not differ in both groups. CONCLUSION: Diltiazem reduced the clinical and electrocardiographical aspects and raises the effort tolerance during the ergometric test in patients with stable angina.

[Effect of ASA on the interaction of von Willebrand factor with the platelet membrane]. / Effetto di ASA sull'interazione del fattore di von Willebrand con la membrana piastrinica.

Ristocetin induces a conformational change on von Willebrand Factor (vWF) similar to that due to the interaction with the subendothelium, by which the former can interact with the Glycoprotein-1 B (GPIB) of the platelet membrane and trigger aggregation and granule content secretion. Platelet Rich Plasma (PRP) treated with Acetyl Salicylic Acid (ASA) loses completely the aggregability induced by addition of Ristocetin whereas ASA-treated and successively Washed Platelets (AWP) supplemented with normal plasma (PPP) give an aggregation and a secretory response to Ristocetin similar to that given by PRP; similarly normal Washed Platelets (WP) supplemented with ASA-treated plasma (APPP) give identical aggregation, and secretion by Ristocetin addition. Ours results indicate that the Ristocetin-vWF complex can trigger two distinct intraplatelet metabolic pathways. A first well known way starts from the activation of Phospholipase A-2 (PL-A2), by which arachidonic acid is produced, that, in turn, undergoes the metabolic pathway leading to Thromboxane A-2; this pathway can be blocked by the intraplatelet ASA by irreversible inactivation of Cyclooxygenase, but it is insensitive to the extra-platelet ASA. A second, independent metabolic pathway, can be triggered by intact vWF, but not by the ASA treated one. It is insensitive to intraplatelet ASA and therefore unrelated to the arachidonic acid metabolism. This pathway could start from the activation of Phospholipase C (PL-C).

[Thoracic duct in CT (author's transl)]. / Il dotto toracico alla TC.

After an anatomical and anatomo-radiological introduction, the authors describe the morphological aspect of the thoracic duct examined by means of lymphangiography performed during computed tomographic examinations.

Analysis of polycystic kidney disease with two new microsatellite markers.

Two (CA) microsatellite polymorphisms have been studied in 209 subjects from Northern Italy, members of 27 Polycystic Kidney Disease families. Polymorphic alleles were analyzed by using a labelled PCR. Results obtained show that these markers highly improve the presymptomatic diagnosis of the disease in subjects at risk. In addition, our findings suggest that a region with a high recombination frequency should exist between PKD1 and SM7.

Expansion of a (CAG)n repeat region in a sporadic case of HD.

The genetic mutation underlying Huntington's disease (HD) has been identified as an expansion and instability of a specific CAG repeat sequence in a gene on chromosome 4. A simple polymerase chain reaction assay has been used for the assessment of the (CAG)n expansion in a 72-year-old woman with typical HD symptoms, but no family history of the disorder. The DNA analysis showed that the patient had an allele with 41 repeat units, in the size range seen in HD chromosomes. Therefore, HD diagnosis is confirmed in this seemingly sporadic case and the disease is newly diagnosed in a large family. The risk of inheriting this unstable expanded allele is discussed. INTRODUCTION--The discovery of an expansion of a trinucleotide (CAG) repeat region in the IT15 gene on the short arm of chromosome 4 has identified the mutational mechanism causing Huntington's disease (HD) and enables the direct diagnosis of affected subjects based on DNA analysis alone. Here a 72-year-old woman with typical HD symptoms, but no family history of the disorder, has been unambiguously diagnosed by using a quick DNA analysis. This is relevant because the disease is newly diagnosed in a large family. MATERIAL AND METHODS--A labelled polymerase chain reaction (PCR) test has been used to amplify the repeat region of the IT15 gene and DNA fragments were analyzed by Polyacrylamide gel electrophoresis. RESULTS--The number the CAG repeats in the proband displayed two alleles of 23 and 41 repeats, respectively. Since normal chromosomes are reported to contain 11-34 repeats, the clinical appearance of HD in the proband is explained by the presence of the repeat expansion. DISCUSSION--The parents of the proposita both died aged over 80 y apparently without neurological signs referable to HD. Hence, this is presumably a sporadic case of the disease. Because of the length of 41 repeats of this HD chromosome, offspring of this proband could inherit the expanded allele with 37 repeats, as expected for the reversal of the trinucleotide expansion. A subject with this intermediate allele could be affected, but would not be affected if the HD IT gene with reduced triplets had recovered its normal function. Thus, in a seemingly sporadic case like the one reported here, despite the PCR analysis, the risk of transmission of HD to her offspring may remain uncertain.

Ionic regulation of endonuclease activity in PC12 cells.

We have investigated the Ca2+ dependency of DNA degradation into nucleosome-sized fragments in intact chromaffin-like PC12 cells and PC12 nuclear fractions. In intact cells we were unable to trigger DNA fragmentation by inducing either transient or sustained elevations of cytoplasmic Ca2+ ([Ca2+]i) with the Ca2+ ionophore ionomycin. On the contrary, DNA fragmentation was induced in intact cells by the intracellular Zn2+ chelator NNN'N'-tetrakis-(2-pyridylmethyl)ethylenediamine (TPEN). To characterize further PC12 cell endonuclease activity, we then investigated digestion by purified PC12 cell fractions of exogenously added plasmids. In nuclear fractions two endonuclease activities were identified: an acidic (pH 5.0) endonuclease activity that was fully Ca2+- and Mg(2+)-independent; and a neutral (pH 7.6) endonuclease activity that was Ca(2+)-independent but Mg(2+)-dependent. Both endonuclease activities were inhibited by Zn2+. Nuclear membrane permeabilization greatly enhanced plasmid digestion at pH 7.6, but not at pH 5.0. This suggests that neutral endonuclease was located in a membrane-bound compartment, whereas acidic endonuclease was freely accessible to the substrate even in the presence of an intact nuclear membrane. In intact nuclei, digestion of genomic DNA could not be triggered by increasing the bivalent cation composition of the medium. On the contrary, in hypotonic medium we observed a large spontaneous nucleolytic DNA degradation that was increased by Zn2+ chelation. However, an acidic pH shift was a potent stimulus for DNA fragmentation in isotonic as well as hypotonic medium.

Simplified construction and characterization of yeast artificial chromosome libraries.

Three yeast artificial chromosome (YAC) libraries were constructed using two human cell lines and the pYAC-RC vector. The main differences from the previously described methods were: i) genomic DNA was digested in low melting point (LMP) agarose blocks with the rare cutting enzyme ClaI; ii) DNA was ligated in melted LMP agarose after agarase treatment; iii) spheroplast regeneration plating was done in calcium alginate thin layer. In addition, a panel of PCR primers was used to identify quickly the presence in the libraries of repetitive and single copy human DNA sequences.