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Biomedical subjects

A Massari

Publications and source records attributed to A Massari.

At least 19 recordsLinked to original sources

In vivo and in vitro exposures of carp and carp tissues to graded concentrations of endocrine disrupting chemicals.

The objective of this study was to correlate morphological and biochemical parameters of reproductive activity in fish exposed to graded concentrations of endocrine disrupting chemicals. Two-year-old carps were exposed for two weeks to graded concentrations of ethynyl-estradiol, tamoxifen, and flutamide. For each chemical, morphological alterations of the gonads and liver were described according to biomarker and sex steroid levels. In parallel, isolated follicular cells (FCs) and testis fragments were incubated in the presence or absence of carp pituitary homogenate and 25-hydroxycholesterol. Media were analyzed for sex steroid level evaluations.

Animals↗

Fish community characterization in two stretches upstream and downstream of the Lambro River confluence with the Po River.

This study aimed to characterize the fish community in two stretches of the Po River, upstream and downstream of the Lambro River confluence, to verify the environmental effects of the presence of endocrine disrupters (EDs). More than 5000 specimens were captured, identified at the species level, and recorded for biometric parameters. Plasma, liver, and gonad fragments were sampled for histological and biochemical analyses. Few specimens belonging to carp and barbel showed gonads with the concomitant presence of male and female tissues and altered plasma levels of vitellogenin and sex steroids; thus, these species will be considered for successive environmental studies.

Animals↗

Genomic analysis of cultivated barley (Hordeum vulgare) using sequence-tagged molecular markers. Estimates of divergence based on RFLP and PCR markers derived from stress-responsive genes, and simple-sequence repeats (SSRs).

Three types of molecular markers have been compared for their utility in evaluating genetic diversity among cultivars of Hordeum vulgare. Restriction fragment length polymorphisms at 71 sites were scored with the aid of probes corresponding to stress-responsive genes from barley and wheat, coding for a low-molecular-weight heat shock protein, a dehydrin, an aldose reductase homolog, and a 18.9-kDa drought-induced protein of unknown function. Indexes of genetic diversity computed in the total sample and within groups of cultivars (two-rowed and six-rowed, winter and spring varieties) indicated high values of genetic differentiation ( F (ST) >15%). A second assessment of genetic diversity was performed by PCR amplification of genomic DNA using as primers 13 arbitrary oligonucleotides derived from sequences of the same stress-responsive genes. A high degree of polymorphism was uncovered using these markers also, but they yielded low values for F (ST) (<7%) among groups of cultivars. Finally, 15 different simple-sequence repeats (AC or AG) were amplified with primers based on unique flanking sequences. Levels of polymorphism and differentiation between groups of cultivars revealed by these markers were quite high. Ordination techniques applied to measures of genetic distance among cultivars demonstrated a remarkable ability of the RFLPs associated with stress-responsive genes to discriminate on the basis of growth habit. The correlation with production data for the cultivars in different environments was also significant. This "functional genomics" strategy was therefore as informative as the "structural genomics" (SSR-based) approach, but requires the analysis of fewer probes.

Base Sequence↗

Histological and histochemical study of female germ cell development in the dusky grouper Epinephelus marginatus (Lowe, 1834).

The developmental stages of female germ cells were analysed in a wild population of the protogynous teleost Epinephelus marginatus (Lowe, 1834). 321 wild dusky grouper females were collected in the South Mediterranean Sea during the spawning season and their ovaries analysed using histological and histochemical techniques. Oocyte morphology, nucleus-cytoplasm ratio (N/C) range, location and movements of cytoplasmic inclusions during primary growth, vitellogenesis and final oocyte maturation were described. The distribution of proteins, lipids and carbohydrates through oocyte development was also investigated in 50 females. Lipid vesicles appeared firstly in the mid ooplasm of oocytes larger than 130 microm, at the beginning of the secondary growth phase. Immediately afterwards, small carbohydrate granules (PAS and Alcian blue positive) appeared before the occurrence of the first yolk granules. Tyrosine-enriched proteins were especially evidenced in the zona radiata interna of late vitellogenic oocytes. Specific lectin binding patterns reflected characteristic differences in the content and distribution of specific sugar moieties expressed in the oocytes during vitellogenesis and final maturation. At the end of vitellogenesis and during final maturation, follicular cells, zona radiata, and cortical alveoli were characterised by a strong increase of specific binding for WGA.

Animals↗

A multicenter evaluation of the CARLA system for the measurement of specific IgE antibodies vs. other different methods and skin prick tests.

The evaluation of specific IgE by using appropriate immunoassays represents a useful alternative diagnostic procedure where skin prick tests (SPTs) are not conclusive in clarifying the etiological role of suspected allergens. This study compares the results of the evaluation of specific IgE by using the CARLA system vs. other commercially available immunoassays (CAP system, Ala-STAT Medical system, ALLERgen IFCI Clone System) carried out on the same blood samples obtained from allergic/SPTs negative patients and vs. SPTs. We evaluated serum specific IgE produced against five selected allergens (Dermatophagoides pteronyssinus, Olea europaea, Parietaria judaica, Lolium perenne and Phleum pratense) by using these immunoassays and the correlations between the results of SPTs and IgE evaluations. We demonstrated a good correlation between these last parameters including a high degree of sensitivity and specificity. The reproducibility of the CARLA system was very high by comparing the results obtained by two different laboratories. The results of the CARLA system were well correlated to those of other well-known immunoassays such as CAP system and Ala STAT system. In conclusion, the CARLA system represents an efficient and reliable immunoassay for the evaluation of serum specific IgE.

Adolescent↗

First observation of intersex cyprinids in the Po River (Italy).

Barbel (Barbus plebejus, Cyprinidae) were captured in the Po River, upstream and downstream from the confluence of the Lambro River, a polluted tributary of the major Italian watercourse. The gonads of the two groups of barbel have been histologically examined, and only the downstream specimens showed histo-morphological alterations that can be related to the Lambro tributary as a source to the main river of endocrine disrupting chemicals, possibly with estrogenic effects. In fact, 50% of the barbel captured (8 of 16 fish) in the downstream reach showed intersex gonads.

Animals↗

A simple sequence repeat-based linkage map of barley.

A total of 568 new simple sequence repeat (SSR)-based markers for barley have been developed from a combination of database sequences and small insert genomic libraries enriched for a range of short simple sequence repeats. Analysis of the SSRs on 16 barley cultivars revealed variable levels of informativeness but no obvious correlation was found with SSR repeat length, motif type, or map position. Of the 568 SSRs developed, 242 were genetically mapped, 216 with 37 previously published SSRs in a single doubled-haploid population derived from the F(1) of an interspecific cross between the cultivar Lina and Hordeum spontaneum Canada Park and 26 SSRs in two other mapping populations. A total of 27 SSRs amplified multiple loci. Centromeric clustering of markers was observed in the main mapping population; however, the clustering severity was reduced in intraspecific crosses, supporting the notion that the observed marker distribution was largely a genetical effect. The mapped SSRs provide a framework for rapidly assigning chromosomal designations and polarity in future mapping programs in barley and a convenient alternative to RFLP for aligning information derived from different populations. A list of the 242 primer pairs that amplify mapped SSRs from total barley genomic DNA is presented.

Chromosome Mapping↗

Normal values of reactive oxygen metabolites on the cord-blood of full-term infants with a colorimetric method.

The main end-point of the study was to evaluate the normal values of reactive oxygen metabolites (ROMs) in healthy full-term babies. Secondary end-points were differences between groups related to modality of delivery, Apgar score, birth weight, gestational age and sex. All apparently healthy babies born at our institution between 8 a.m. and 8 p.m. Monday to Friday with gestational age 37-42 weeks, delivered both vaginally or by caesarean section and without foetal distress and perinatal asphyxia. ROMs were evaluated by a colorimetric method (d-ROM test) on cord-blood immediately after birth. The values are reported as arbitrary unit U. Carr. Statistical analysis was performed by t-test and by multiple and stepwise regression analysis. We have analyzed 80 babies with mean birth weight 3301 +/- 446 g. and mean gestational age 39.5 +/- 1.0 weeks. The male:female ratio was 1.56 and the median (range) Apgar score was 9 (7-10) at 1' and 10 (9-10) at 5'. The babies born by vaginal delivery were 37 out of 80 while the remaining 43 were delivered by cesarean section. Because the two groups did not differ for the clinical characteristics they were considered together for the determination of the mean value of ROMs and indicated as "total". The mean value +/- SD of ROMs of the "total" was 115.5 +/- 32.6 U. Carr. Significant differences in the mean value of ROMs were not found related to type of delivery, birth weight, gestational age, and Apgar score at 1' and 5'. Instead the female infants had a significantly lower mean value of ROMs than the male babies (respectively 104.4 +/- 32.2 vs 120.2 +/- 30.6 U. Carr.; p = 0.031). Multiple and stepwise regression analyses both demonstrated that the sex of the neonate is able to independently influence the value of ROMs (respectively p = 0.025 and p = 0.035). The main end-point of the study was to determine the standard reference values for this method in the healthy full-term infant at birth: the values of ROMs we found in the "total" population are lower than those of healthy adults (between 250-300 U. Carr.) and similar to those of adults treated with steroids or antioxidant drugs. The finding that the female sex is able to independently determine lower values of ROMs at birth compared to the male sex, lets speculate that the female infants are less prone to oxidative stress in the first moments of life.

Colorimetry↗

Evaluation of reactive oxygen metabolites with micromethod in neonates: determination of standards of normality in full-term babies.

We have evaluated the value of reactive oxygen metabolites (ROMs) in 98 full-term neonates at 72 +/- 6 hours of life with a new colorimetric simple and rapid method (d-ROMs Test, Diacron s.r.l.) to establish the normal values for infants. The mean value of ROMs we have obtained in the total of the population was 127.9 +/- 39.2 U.Carr. We have then considered subgroups of infants on the basis of vaginal delivery vs cesarean section, and asphyxia vs non asphyxia during the delivery. We have found no difference within these groups and between the subgroups and the total population for the value of ROMs. We have preferred to consider as reference value for normality that of babies without intrapartum asphyxia, independently of the mode of delivery. This value is 125.2 +/- 27.6 U.Carr. The value of normality for full-term infants is lower than that reported for adults (between 250-300 U.Carr). This can be interpreted as a particular response of full-term baby to oxidative stress.

Asphyxia Neonatorum↗

Non-invasive early prenatal molecular diagnosis using retrieved transcervical trophoblast cells.

Fetal DNA was recovered from 17 of 39 (44%) transcervical cell (TCC) samples obtained between 7 and 9 weeks of gestation by endocervical canal flushing. Trophoblast retrieval was adequate for polymerase chain reaction (PCR) amplification of Y chromosome-specific DNA sequences and detection of paternal-specific microsatellite alleles. The fetal sex predicted by PCR in TCCs was confirmed in all cases by karyotype analysis of chorionic villi at 10 weeks of gestation. The absence of the disease-associated paternal alleles in TCC samples from two pregnancies at risk for spinal muscular atrophy and myotonic dystrophy predicted unaffected fetuses in agreement with subsequent results on chorionic villi and newborns' leukocytes. A trisomy 21 fetus was diagnosed in TCCs using fluorescent in situ hybridization (FISH) and semi-quantitative PCR analysis of superoxide dismutase-1 (SOD 1). Present experience indicates that TCC sampling is a promising technique for early prenatal monitoring of Mendelian disorders and chromosome aneuploidy.

Adult↗

Postzygotic instability of the myotonic dystrophy p[AGC] in repeat supported by larger expansions in muscle and reduced amplifications in sperm.

We have analysed the [AGC] expansion in leucocytes, muscle and sperm from 17 individuals affected by myotonic dystrophy (DM). Skeletal muscle showed a larger repeat number than leucocytes in the same patient. A similar degree of expansion was detected in differently affected muscles of a single patient. The germline mutation (< or = 350 repeats) was expanded in somatic cells of the progeny in all patients examined. Our results provide evidence of an early postzygotic instability of the [AGC] repeat in DM.

Adolescent↗

[Effect of 2 types of diet changes on dietary habits, body weight and cholesterol levels in high risk cardiovascular patients].

Several non-pharmacological interventions such as weight reduction in obese subjects or diet alteration in subjects having hypercholesterolemia have been shown to be effective in therapeutic trials. Our aim was to test the value of two different ways of teaching patients about their diet. From March 1, 1993, to May 30, 1994, 300 consecutive patients seen in a one-day care hospital were randomised into two groups. The 2 inclusion criteria were: 1) body mass index > 27 kg/m2 in men and > to 25 kg/m2 in women and/or 2) presence of a hypercholesterolemia defined by a total cholesterol > 6.5 mmol/l. Patients in the first group (C) were educated in a 20 to 50 minutes consultation tailored to their needs. Patients in the second group (CC) were given in addition a one-hour course about diet. The goal of the diet was to loss at least 3 kg of body weight and/or to have a cholesterol value below 6.5 mmol/l without treatment. All Patients were followed-up by the same 3 dietician nurses. An out-patient visit was planned at 3 months, and a recall letter was sent to the patients who missed their appointments. Among the 300 patients, 169 (55%) were seen at the 3-month outpatient visit. This proportion did not differ between the 2 groups. Knowledge on diet was assessed by the same 33-item self-administered questionnaire. At baseline scores were comparable between groups (16 vs 17). Scores improved more in the CC than in the C group both at the end of the teaching question (27 vs 23 in the CC and C group respectively, p < 0.001), and at 3 months (25 vs 23 in the CC and C group respectively; p < 0.001). Total cholesterol decreased below 6.5 mmol/l in 28% of the patients with dyslipidemia and a weight loss > 3 kg was observed in 32% of the obese subjects, but improvement did not differ between the 2 groups. We conclude that a specific one-hour course on diet is able to improve knowledge of patients more than a consultation alone, but that better knowledge did not result in improved alteration of risk factors at 3 months.

Adult↗

Presence of angiotensin II immunoreactivity in the ovary of the rainbow trout, Oncorhynchus mykiss.

Renin-angiotensin system is present in mammalian ovarian follicular fluid and follicles. The demonstration of specific binding sites for angiotensin II in the follicular structures of rats suggests that angiotensin II may be related to the regulation of steroid biosynthesis in the mammalian ovary. Little is known about the presence and the action of angiotensin II in nonmammalian ovaries. An immunocytochemical investigation of angiotensin II in developing ovarian follicles of trout has been carried out. The specific antiserum was raised against (Val5)-angiotensin II of the trout. There was strong immunoreactivity in the ooplasm of endogenous vitellogenic follicles. Angiotensin II immunoreactive material was present in the follicular cells, but not within oocytes at the end of vitellogenesis. Immunocytochemically angiotensin II-like material in the ovary of rainbow trout fluctuates during the cycle of development.

Angiotensin II↗

Low-molecular-weight sodium hyaluronate in the treatment of bacterial corneal ulcers.

A double-blind clinical trial was performed on 26 patients suffering from corneal ulcers of proven (i.e., culture-positive) bacterial etiology. After their recruitment, the subjects were randomly assigned to one of the following treatment protocols: (1) tobramycin (15 mg/ml) in saline applied at 1 drop/h or (2) tobramycin (15 mg/ml) in low-molecular-weight hyaluronic acid applied at 1 drop/h. The sample size was adjusted according to a type I error of 0.01 and type a II error of 0.05 for a minimal expected difference of 35%. The healing time was calculated from the beginning of treatment to the day on which a follow-up fluorescein test proved to be negative. The mean healing time (+/- SD) was 3.5 +/- 0.9 days in the sodium hyaluronate group and 5.9 +/- 1.5 days in the saline group (P less than 0.001). These results suggest that treatment with an antibiotic dissolved in low-molecular-weight sodium hyaluronate can further shorten the clinical course of a bacterial corneal ulcer.

Adolescent↗

Atrial natriuretic peptide in trout ovarian follicles.

An immunocytochemical study on atrial natriuretic peptide (ANP) was carried out in trout ovaries using an antiserum raised against human atrial natriuretic peptide (hANP). Weakly immunoreactive material was observed both in the ooplasm of the previtellogenic and the early vitellogenic oocytes and in the follicular cells of the late vitellogenic oocytes. Binding sites for hANP have been located using 125I-ANP and light-microscopic autoradiography. These studies provide evidence of specific binding of ANP-like material in both early and advanced vitellogenic oocytes. Autoradiograms indicated that the ANP-like binding sites were localized to a subpopulation of follicles. The presence of ANP-like immunoreactive material and its specific binding sites in the ovary suggest a role for atrial natriuretic peptide as a regulator of the fish ovarian function.

Animals↗