Stem cell transplantation for primary immunodeficiencies.

BMT is curative in almost 75% of children affected by severe primary immunodeficiencies (PIDs). Recently, the chance of cure has increased thanks to the availability of matched unrelated donors (MUDs). Nevertheless, besides the conventional indications to BMT (profound or absent T-cell function, profound or absent natural killer function, known syndromes with T-cell deficiencies), indications to BMT for PIDs affecting the quality of life or having an expectation of life that does not exceed the third-fourth decade remain unclear. Infact, if it is evident that the survival rate in an infant grafted for a PID with a MUD is expected to be more than 80%, alternative treatments such as gene therapy are now available.

Short-term treatment with mycophenolic acid and tacrolimus is tolerogenic for INS-1 cell clone transplantation and the deleterious effects of the drugs are limited: in vivo and in vitro studies.

One of the major requirements for a successful and life-lasting organ transplant is the access to safe, least toxic and permanent tolerance-inducing drugs. In this study we wished to evaluate the effects of tolerogenic doses of the immunosuppressive drugs mycophenolic acid (MPA) and tacrolimus (Tac) on clonal beta-cell lines, both in vivo and in vitro. Here we demonstrate that combined administration of low-dose MPA and Tac for 23 days induced permanent tolerance in an allogeneic beta-cell line transplant in Wistar rat liver through the portal vein. This short-term treatment of tolerogenic doses of the two drugs was deleterious to the survival of the transplanted cells but a small percentage of the cells could resist the effect and become fully active when the drugs were removed. The surviving cells, retrieved from growth in vivo, did not exhibit increased resistance in comparison to the original cells when tested in vitro at two glucose concentrations, 10 and 20 mM. The presence of a small percentage of resistant cells at the two glucose concentrations was also detected in the in vitro study after a continuous 8-day treatment demonstrating that the in vivo resistance was not related to micro-environmental protection but possibly to a phenotypic cell state that is yet to be determined.

[Controlled release vaginal dinoprostone for the induction of labour]. / Induzione del travaglio di parto con dinoprostone vaginale a rilascio controllato.

AIM: To study the efficacy and safety of controlled release dinoprostone (PGE2) for the induction of labour. METHODS: From March 2001 to August 2002, a total of 173 patients hospitalized for postdate pregnancy, maternal hypertension, intrauterine delayed growth, reduction of amniotic fluid, were studied. In 33 of these patients, a hydrogel strip containing 10 mg dinoprostone, inserted into the posterior fornix of the vagina and release PGE2 at constant rate of 0.3-0.4 mg/h was used. RESULTS: Indications for induction, mode and time of delivery, maternal, fetal and neonatal parameters anomalies are evaluated. In 23 patients dinoprostone was used for postdate pregnancy, in 4 for maternal hypertension, in 4 for oligohydramnios and in 2 for intrauterine delayed growth. In 2 cases, the vaginal insert was removed for hyperstimulation in 1 case and for abnormal fetal heart rate in the other case. After vaginal delivery, Apgar scores and umbilical artery pH values were evaluated without anomalies. Vaginal delivery within 24 hours occurred in 51.6% of patients and medium time to delivery was 17.5 (nulliparous) and 12.3 (multiparous) hours. Only one case of uterine hyperstimulation to PGE2 was recorded, and the induction was suspended. The cesarean section rate was 12% and vaginal delivery after 24 hours was 38.7%. CONCLUSION: The continuous release of PGE2 from the vaginal insert permits a controlled and safe induction of labour, minimal risk of hyperstimulation, fast and easy removal of drugs in cases of fetal or maternal anomalies, easy accetability for patients, but high cost.

Use of electrical conductivity of amniotic fluid in the evaluation of fetal lung maturation.

The aim of this work is to evaluate, by means of an alternative method, the fetal lung maturation by means of measurement of the electrical conductivity of the amniotic fluid correlated with the phospholipid content. In vitro measurements were made in some 100 amniotic fluid samples at different gestational stages. A considerable change was observed in the electrical conductivity of the amniotic fluid in the last period of pregnancy, which reflects the increase in phospholipid concentration, when lung maturation is reached. We hope that further investigations, on wider case histories, will enable the application in vivo of the above-mentioned methodology.

Gene therapy vectors based on adeno-associated virus type 1.

The complete sequence of adeno-associated virus type 1 (AAV-1) was defined. Its genome of 4,718 nucleotides demonstrates high homology with those of other AAV serotypes, including AAV-6, which appears to have arisen from homologous recombination between AAV-1 and AAV-2. Analysis of sera from nonhuman and human primates for neutralizing antibodies (NAB) against AAV-1 and AAV-2 revealed the following. (i) NAB to AAV-1 are more common than NAB to AAV-2 in nonhuman primates, while the reverse is true in humans; and (ii) sera from 36% of nonhuman primates neutralized AAV-1 but not AAV-2, while sera from 8% of humans neutralized AAV-2 but not AAV-1. An infectious clone of AAV-1 was isolated from a replicated monomer form, and vectors were created with AAV-2 inverted terminal repeats and AAV-1 Rep and Cap functions. Both AAV-1- and AAV-2-based vectors transduced murine liver and muscle in vivo; AAV-1 was more efficient for muscle, while AAV-2 transduced liver more efficiently. Strong NAB responses were detected for each vector administered to murine skeletal muscle; these responses prevented readministration of the same serotype but did not substantially cross-neutralize the other serotype. Similar results were observed in the context of liver-directed gene transfer, except for a significant, but incomplete, neutralization of AAV-1 from a previous treatment with AAV-2. Vectors based on AAV-1 may be preferred in some applications of human gene therapy.

Adeno-associated virus as a vector for liver-directed gene therapy.

Factors relevant to the successful application of adeno-associated virus (AAV) vectors for liver-directed gene therapy were evaluated. Vectors with different promoters driving expression of human alpha-1-antitrypsin (alpha-1AT) were injected into the portal circulation of immunodeficient mice. alpha-1AT expression was stable but dependent on the promoter. Southern analysis of liver DNA revealed approximately 0.1 to 2.0 provirus copies/diploid genome in presumed head-to-tail concatamers. In situ hybridization and immunohistochemical analysis revealed expression in approximately 5% of hepatocytes clustered in the pericentral region. These results support the use of AAV as a vector for diseases treatable by targeting of hepatocytes.

Aggressive behavior as a function of Taekwondo ranking.

This study examined the effect taekwondo training has on the level of aggression of children between the ages of 6 and 11 yr. 68 students (boys and girls) were tested using a behavior checklist designed to measure children's aggression. Analysis indicated a significant inverse relationship between the children's taekwondo rank and their aggression.

Thyroid and steroid receptors.

Estradiol and progesterone receptor proteins (ER, PgR) have been demonstrated in neoplastic and non-neoplastic human thyroid. The aim of this study was to determine the sexual steroid receptor content of pathological non-malignant thyroid (solitary adenoma, simple nontoxic goiter), and the adjacent normal tissue of the thyroid. The results show the presence of ER and PgR (cytosolic and/or nuclear) in most of examined tissues both pathological and normal. Low levels of steroid receptors are found in the cytosol fraction, whereas the receptor content is higher in the nuclear fraction. No correlation could be found between receptor levels and patients' age, menstrual state in females, and sexual steroid circulating hormones. A significant difference has been observed between adenomas and simple goiters (p less than 0.001) in the nuclear ER, and between normal tissue surrounding adenoma and normal tissue in goiters (p less than 0.05). This result suggests that estradiol and progesterone receptors are present in thyroid tissue and may have a physiological function.

Estrogen and progesterone receptors in uterine leiomyomas.

The purpose of our study was to evaluate the contrasting data between estrogen receptors (ER) and progesterone receptors (PgR) in uterine leiomyomas, endometrium and myometrium. Uteri were obtained from 30 women undergoing hysterectomy for uterine leiomyomas (24 premenopausal and 6 postmenopausal patients). Cytosolic and nuclear ER and PgR were analysed in uterine tissues by the method of single saturation point assay at a final concentration 5 nM of [3H-17 beta]estradiol and [3H]ORG-2058. Our results confirm that leiomyomas undergo changes similar to those observed in other uterine tissues during the menstrual cycle. The ER levels show decreased values during the postovulatory phase. After subdividing leiomyomas according to their topographic criteria, as submucous or subserosal, we observed that ER and PgR are significantly more numerous in submucous than in subserosal myomas, both in proliferative and in the secretive phase of the cycle. These results agree with the hypothesis that leiomyomas have a different etiology and may explain why some myomas decrease after progestin therapy (submucous), while others (subserosal) remain unchanged.

Relationship between PRL levels and steroid receptors in breast cancer.

The role of PRL in human breast cancer is still debated. We examined 259 women undergoing surgery for breast cancer (K = 183) and for other diseases (n = 76). In all subjects blood samples were taken for the RIA assay of PRL, during the preanaesthesia. In the breast cancer specimens, estrogen (ER) and progesterone (PGR) receptors were assayed and T.N.M. was evaluated. PRL mean values were assayed and T.N.M. was evaluated. PRL mean values were found to be above the normal range in both groups. PRL values in ER+ and PGR+ were lower than in ER- and PGR-. We also divided the patients into two groups (PRL values greater than 50 or less than 50 ng/ml). The difference between the two groups was significant for ER (p less than 0.005) and PGR (p less than 0.05), using Student's unpaired t-test. Besides this, we observed a significant negative linear correlation between PRL values and receptor values. The finding of this correlation suggests that high PRL values could be associated with unfavourable prognosis in breast cancer.

Epidemic campylobacteriosis associated with a community water supply.

In May 1983, an estimated 865 cases of epidemic gastrointestinal disease occurred in Greenville, Florida. Surveillance of pharmacy sales of antidiarrheal medicines suggested that the outbreak was confined to Greenville and its immediate vicinity. Surveys demonstrated that the gastrointestinal illness attack rates inside and outside the city limits were 56 per cent (72/128) and 9 per cent (7/77), respectively (relative risk (RR) = 6.2); consumption of city water was associated with illness (RR = 12); and as water consumption increased, the attack rate also increased (p less than 0.001). Four adults were hospitalized and one outbreak-related case of Guillain-Barre' syndrome was identified. Campylobacter jejuni was isolated from specimens from 11 ill persons; serologic studies showed the development of Campylobacter-specific antibodies. Fecal coliforms were found in water samples, but Campylobacter was not recovered from water. The city water plant, a deep well system, had numerous deficiencies including an unlicensed operator, a failure of chlorination, and open-top treatment towers. Birds were observed perching on the open-top treatment tower. Of 38 birds trapped seven weeks later, 37 per cent harbored C. jejuni; however, plasmid and serotyping studies showed that strains were not the same as the common strain from ill persons. This outbreak suggests that water systems that are unprotected from contact with birds may become contaminated and a source of outbreaks of human campylobacteriosis.