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Biomedical subjects

L Molteni

Publications and source records attributed to L Molteni.

At least 19 recordsLinked to original sources

Increased intestinal permeability precedes clinical onset of type 1 diabetes.

AIMS/HYPOTHESIS: Recent observations have shown subclinical intestinal abnormalities in human type 1 diabetes. Whether these are related to the pathogenetic process or secondary to the diabetes remains to be clarified. The aim of this study was to investigate this issue by examining intestinal permeability to sugars in subjects at different stages of type 1 diabetes: preclinical, new-onset and long-term established disease. METHODS: Eighty-one subjects with islet autoimmunity (18 preclinical, 28 new-onset and 35 long-term type 1 diabetes) and 40 healthy control subjects were investigated by a lactulose-mannitol test, consisting of oral administration of the two sugars and measurement of their urinary excretion. RESULTS: All groups of subjects with islet autoimmunity showed an increase in intestinal permeability (p < or = 0.009 vs controls) to the disaccharide lactulose, indicative of a damaged barrier, but a similar permeability to the monosaccharide mannitol (NS vs controls), indicative of an integral surface mucosa; consequently there was an increase in the lactulose:mannitol excretion ratio (p < or = 0.025 vs controls). CONCLUSIONS/INTERPRETATION: These findings indicate the presence of a subclinical enteropathy associated with type 1 diabetes that is already detectable before clinical onset of the disease, and suggest that the small intestine is an organ participating in the pathogenetic process of type 1 diabetes.

Adolescent↗

Cattle rob(1;29) originating from complex chromosome rearrangements as revealed by both banding and FISH-mapping techniques.

Sixteen carriers of rob(1;29) (one of which was homozygous) from six different breeds (four Italian and two Portuguese), two heterozygous carriers of rob(26;29), three river buffaloes and two sheep were cytogenetically investigated in this study by using banding and FISH-mapping techniques (the latter only in cattle and river buffalo). Single- and dual- colour FISH were used with bovine probes containing both INRA143 (mapping proximally to BTA29) and bovine satellite (SAT) DNA SAT I, SAT III and SAT IV (mapping at the centromeric regions of cattle chromosomes). The combined use of these probes, the comparison of rob(1;29) with the dicentric rob(26;29) and with both river buffalo and sheep chromosomes (biarmed pairs) allowed us to hypothezise that rob(1;29) originated from complex chromosomal rearrangements through at least three sequential events: (a) centric fusion with the formation of a dicentric chromosome; (b) formation of a monocentric chromosome with loss of SAT I from both BTA1 and BTA29, most of SAT IV from BTA29 and, probably, some repeats of SAT III from BTA1; (c) double pericentric inversion or, more probably, a chromosome transposition of a small chromosome segment containing INRA143 from proximal p-arms to proximal q-arm of the translocated chromosome.

Animals↗

Fertility of cryopreserved sperm in three bulls with different Robertsonian translocations.

The fertility of three bulls carrying different Robertsonian translocations (rob(1;29), rob(14;17) and rob(26;29)) was evaluated. Oocytes-cumulus complexes obtained from slaughterhouse-derived ovaries were matured and then fertilised in vitro with frozen/thawed seminal material from the above mentioned subjects, and from control bulls with normal karyotype. An assessment was first made of the concentration, vitality and acrosome integrity of the seminal material to be sure that possible differences in the results of the in vitro fertilisation experiments were not due to seminal material quality. The results of the experiments, evaluated by the percentage of cleaved embryos and blastocysts per cleaved embryo, indicated that the three bulls carrying Robertsonian translocations had similar fertilising power and semen qualitative parameters to the controls. These data suggest that neither gametogenesys impairment nor decreased spermatozoa fertilising capacity is responsible for the reduced fertility in bulls with Robertsonian translocations. What the data do confirm is that the observed in vivo hypofertility for karyologically abnormal bulls is mainly due to early embryonic mortality.

Animals↗

Comparative FISH-mapping of the survival of motor neuron gene (SMN) in domestic bovids.

A comparative fluorescence in situ mapping of the SMN gene was performed on R-banded chromosome preparations of cattle (Bos taurus, BTA, 2n = 60), river buffalo (Bubalus bubalis, BBU, 2n = 50), sheep (Ovis aries, OAR, 2n = 54) and goat (Capra hircus, CHI, 2n = 60), as well as on those of a calf from Piedmont breed affected by arthrogryposis. SMN was located on BTA20q13.1, OAR16q13.1, CHI20q13.1 and BBU19q13. These chromosomes and chromosome bands are believed to be homeologous, confirming the high degree of chromosome homeologies among bovids. The position of SMN was refined in cattle, compared to the two previous localizations, while it is a new gene assignment in the other three bovids. A comparative fiber-FISH performed on extended chromatin of both normal cattle and calf affected by arthrogryposis revealed more extended FITC signals in the calf, compared to the normal cattle (control), suggesting a possible duplication of the SMN gene in the calf affected by arthrogryposis. .

Animals↗

A new balanced autosomal reciprocal translocation in cattle revealed by banding techniques and human-painting probes.

Three hundred and twenty-two (264 males and 58 females), randomly sampled Grey Alpine cattle individuals from Northeastern Italy, were investigated cytogenetically by both conventional chromosome staining and R-banding. Two hundred and eighty-one (87%) individuals had a normal karyotype and 41 (13%) carried chromosomal aberrations such as (a) rob(1;29) in two individuals, (b) rob(26;29) in 36 individuals, (c) XX/XY-chimerism in two individuals, and (d) an abnormally long chromosome in one individual. All these aberrations except (d) have been described before. GBG-, RBG-, CBA-banding and sequential GBG/CBA- and RBG/CBA-banding techniques revealed that the abnormally long chromosome was the result of a reciprocal translocation between chromosomes 1 (q21-->qter) and 5 (q11-->q33), as confirmed also by chromosome painting with human chromosome 3 and 12 probes. The dam of the carrier bull carried the same translocation, while the grandam showed a normal karyotype. Since the sire of the dam was not available for study, no conclusion about the origin of the chromosome translocation could be drawn. The carrier bull was eliminated because of poor fertility. The dam had three other calves, which all were chromosomally normal. On average the dam had to be served 2.5 times (breed average was 1.2) to be in calf.

Animals↗

A case of azoospermia in a bull carrying a Y-autosome reciprocal translocation.

During normal cytogenetic investigations on the Chianina cattle (BTA) breed, a normal looking young bull was found to carry an abnormal Y chromosome which was a product of a reciprocal translocation between chromosomes Y and 9. This was revealed by both CBA- and RBG-banding techniques and was clearly confirmed by FISH-mapping analysis with IDVGA50 (which paints the complete Yq arm in a normal Y), as well as with AMD1, CGA, IGF2R (mapping to BTA9q16, BTA9q22 and BTA9q27-->q28, respectively) and SRY (mapping to normal BTAYq23). Analysis on sperm from four different samples revealed azoospermia in the carrier, indicating that the rcp(Y;9) induces sterility in the bull.

Animals↗

Immortalized luteinizing hormone-releasing hormone neurons show a different migratory activity in vitro.

The development of two cell lines (GT1 and GN) of immortalized LHRH neurons has allowed an accurate study of the mechanisms controlling the synthesis and the secretion of LHRH. These cell lines, obtained in mice by genetic targeted tumorigenesis, retain many of the phenotypic characteristics of LHRH neurons. Of interest, GT1 cells derive from an hypothalamic tumor, whereas GN cells were obtained from a tumor localized in the olfactory bulb. The different origin of these cell lines lead to hypothesize that they might represent hypothalamic postmigratory neurons (GT1 cells), or LHRH neurons blocked at an early stage of their migration (GN cells). Using different experimental procedures, we found that the two cell subclones GT1-7 and GN11 express a different morphology and migratory behavior in vitro. In particular, we found that GN11 cells, but not GT1-7 cells, show the morphological shape of migrating neurons. When analyzing the spontaneous motility we found that only GN11 cells express a high capacity of migrating in a matrix of collagen gel. Moreover, in a chemomigratory assay GN11 cells did show a significant response to the chemotactic stimulus represented by the FBS. On the contrary, GT1-7 cells show very low spontaneous motility and appear insensitive to the FBS stimulus. These results suggest that the simultaneous use of the GT1-7/GN11 cells may represent an experimental tool for screening the factors possibly involved in the control of the migratory processes of LHRH neurons in normal and in pathological conditions, such as those due to their impaired migration, like it happens in Kallmann's syndrome.

Animals↗

New cases of XXY constitution in cattle.

The present paper describes two cases of an XXY condition in Chianina cattle. Both young bulls were routinely investigated cytogenetically before entering progeny test stations. Every cell examined in the blood cell cultures showed an XXY constitution. The histological study of the gonads, performed on only one bull, showed degradation of the seminiferous tubules. Only Sertoli cells and hyperplastic interstitial cells were observed.

Animals↗

A new centric fusion translocation in cattle: rob (13;19).

A new Robertsonian translocation has been found in cattle. A bull from Marchigiana breed (central Italy) was found to be a heterozygous carrier of a centric fusion translocation involving cattle chromosomes 13 and 19 according to RBA-banding and cattle standard nomenclatures. CBC-banding revealed the dicentric nature of this new translocation, underlining the recent origin of this fusion. In fact, both the bull's parents and relatives had normal karyotypes. In vitro fertilization tests were also performed in the bull carrying the new translocation, in two bulls with normal karyotypes (control) and in four other bulls carrying four different translocations.

Animals↗

Genetic analysis of 38XX males with genital ambiguities and true hermaphrodites in pigs.

In pig, the frequency of intersexuality ranges from 0.1 to 0.6%, depending on the breed. In a closed pig herd at INRA an intersex condition was observed in 0.75% of 'females'. The present study describes 11 animals with a 38XX karyotype and the presence of testicular tissue. Phenotypically, all presented with abnormal external or/and internal genitalia. Southern blot analysis with Y-specific probes (SRY and ZFY) revealed the absence of Y material in all animals tested. By polymerase chain reaction (PCR) amplification, 10 of 11 intersex pigs lacked the SRY gene in gonad DNA. These data are compatible with an autosomally (or pseudoautosomally) determined mechanism. Moreover, analysis of familial cases seemed to indicate that 38XX male pseudohermaphrodites and 38XX true hermaphrodites may represent alternative manifestations of the same genetic defect.

Animals↗

The hypertrophied myocardium and coronary disease. Structural changes in patients submitted to aortocoronary bypass surgery.

Seventeen patients with coronary disease submitted to myocardial revascularization were studied. Ten patients had a hypertrophied ventricle, and 7 had normal ventricular mass. Myocardial biopsies were obtained before ischemia and at the time of reperfusion and were assessed for: volume fraction of fibrous tissue, myocyte diameter, morphometric mitochondrial studies and ultrastructural changes. The volume fraction of fibrous tissue in patients with hypertrophied ventricle was 1.9 +/- 0.04, and in patients with normal ventricular mass was 0.9 +/- 0.01 (p less than 0.05). The diameter of the myocyte was 23 +/- 0.3 microns and 18 +/- 1.2 microns for patients with hypertrophied and normal ventricular mass, respectively (p less than 0.01). The value of volumetric density for pre-ischemia samples in patients with a hypertrophied ventricle was 23 +/- 2.2 and in patients with normal ventricular mass was 35 +/- 2.7 (p less than 0.02). Grades 3 and 4 of damaged mitochondria were significantly increased in reperfusion samples from patients with a hypertrophied ventricle compared to pre-ischemia samples. Collagen growth was increased in hypertrophied hearts which were also more sensitive to the ischemia/reperfusion mechanism.

Biopsy↗

The value of the echo-Doppler in cardiomyoplasty procedures.

UNLABELLED: Echo-Doppler determinations were performed in patients submitted to cardiomyoplasty procedure using the latissimus dorsi muscle graft (LDMG). In four patients who had 1 year of follow-up or more the left ventricular internal dimension in diastole (LVIDd), left ventricular internal dimension in systole (LVIDs), septal-E point separation (E-septum), distance of the mitral valve (E-E'), maximal aortic cuspid separation (MACS) and left atrium dimension (LA) were determined by two-dimensional echocardiogram. With a pulsed Doppler in the aortic root the following parameters were obtained: aortic peak flow (AoPF), velocity time integral (VTI), and ejection period (EP). With the stimulator on, the determinations were done using a ventricle muscle (VM) delay of 25, 75, and 250 msec. RESULTS: No significant differences were observed by Echo determinations between the stimulator off and on. With the supporting contraction of the LDMG the best hemodynamic performance was obtained when the VM delay ranged between 75 and 250 msec. Values with the stimulator off and on were as follows: AoPF: 90.8 +/- 8 cm/sec and 104.5 +/- 9 cm/sec; VTI: 14.8 +/- 2 cm and 19.2 +/- 2 cm; EP: 230 +/- 10 msec and 245 +/- 20 msec. A decrease in the systolic pressure of the pulmonary artery and a decreased functional regurgitation of the mitral and tricuspid valve were observed in the follow-up determinations. All patients showed restricted mitral flows that remained unchanged during the follow-up. CONCLUSIONS: In this group of patients the echo-Doppler showed an improvement of the systolic function of the left ventricle when it is supported by the LDMG.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Clinical experience with dynamic cardiomyoplasty.

Dynamic cardiomyoplasty using the latissimus dorsi muscle graft (LDMG) was applied to 11 patients with dilated cardiomyopathy of different etiologies. Our first case was operated on July 1987. All patients were in Functional Class (FC) IV New York Heart Association (NYHA) and had multiple previous admissions in intensive care units for intractable cardiac insufficiency. Two patients died during the procedure, the perioperative mortality was 18%. One patient died of recurrent acute myocarditis at the fourth postoperative month and a second patient died in the sixteenth postoperative month. Both patients were asymptomatic, in FC I-II, and the cause of death was sudden arrhythmia (ventricular fibrillation). The remaining seven patients (63.6%) were in FC I-II and the longest follow-up was 30 months. In all patients, the ejection fraction (EF) by radioisotopic ventriculography improved an average of 50% as well as their ergometric test. Dynamic cardiomyoplasty appears to be a satisfactory alternative for the treatment of patients with severe cardiac insufficiency secondary to dilated cardiomyopathy. Its main advantage seems to be a significant improvement in the quality of life for these patients.

Adult↗

Effect of supplementing cardioplegic solution with deferoxamine on reperfused human myocardium.

Fourteen randomized patients undergoing myocardial revascularization were divided into group A standard hypothermic cardioplegic solution) and group B (the same cardioplegic solution supplemented with deferoxamine 1000 mg/L). In all patients myocardial biopsy specimens were obtained before ischemia and during reperfusion and were assessed for chemiluminescence (to indirectly determine oxygen-free radical activity) and for electron microscopic studies. Chemiluminescence in group A showed a photoemission of 36.5 +/- 1.5 cpm/mg protein X10(-3) for the preischemia samples and 72 +/- 5.7 cpm/mg protein X10(-3) for the reperfusion samples (p less than 0.01). In the patients who received deferoxime (group B), values for chemiluminescence for preischemia and reperfusion samples were not significantly different. Electron microscopic studies showed a significant increase in grade 4 (severely damaged) mitochondria in reperfusion biopsy specimens from both groups as compared with preischemia samples. However, reperfusion samples from group B showed a better preservation of myocardial cells with marked reduction of grade 4 (severely damaged) mitochondria. These results support the hypothesis that oxygen-free radicals are responsible in part for the production of reperfusion injury in the human heart. They suggest that this mechanism may be at least partially controlled by adding an iron chelating agent such as deferoxime.

Cardioplegic Solutions↗

Reduction of reperfusion injury with mannitol cardioplegia.

Forty consecutive patients undergoing myocardial revascularization were divided into two equal groups: group 1 received standard cardioplegic solution, and group 2 received a solution containing mannitol, 59.8 mmol/L. In 6 patients in each group, myocardial biopsies were done before ischemia and at the time of reperfusion. Samples were assessed by chemiluminescence to determine oxidative stress and by electron microscopic studies. A significant reduction in atrial arrhythmias was observed in the mannitol group. Chemiluminescence in group 1 showed a photoemission of 37.6 +/- 3.5 cpm/mg of protein x 10(-3) for the preischemia samples and 74.8 +/- 16 cpm/mg of protein x 10(-3) for the reperfusion samples (p less than 0.001). In group 2, the values for chemiluminescence were 37.7 +/- 3.4 cpm/mg of protein x 10(-3) and 40 +/- 6.1 cpm/mg of protein x 10(-3), respectively (p = not significant). Electron microscopic studies showed, for group 1, increased grades of damaged mitochondria in the reperfusion biopsy specimens compared with the preischemia biopsy specimens (p less than 0.01). In group 2, differences for damaged mitochondria were not significant. These results support the hypothesis that mannitol reperfusate significantly reduces myocardial damage in patients undergoing open heart procedures. They also suggest that this protective effect may be in part secondary to the antioxidant property of mannitol, although other mechanisms may have accounted for or contributed to the improved outcome after ischemia.

Biopsy↗

Synaptonemal complexes analysis in a bull carrying a 4;8 Robertsonian translocation.

Synaptonemal complexes analysis was performed using electron microscopy on surface-spread spermatocytes of a bull heterozygous for the 4;8 Robertsonian translocation. In 19 cells examined, the longest autosomal complex showed kinetochores in a central position whereas the remaining autosomal complexes showed terminal kinetochores. Synapsis in the trivalent appeared complete in all cells, and the trivalents usually showed a CIS configuration. The arm ratio varied from 1.05 to 2.04 with an average of 1.32 +/- 0.43. Out of 47 cells showing X-Y bivalents, 34 showed a small synaptonemal complex at one extremity of the X chromosome, and an unstained gap in the Y chromosome. There was no association between the X-Y bivalent and the trivalent. The absence of association would explain the normal spermatogenesis noted in this bull, in contrast to human and mouse carriers of translocations which show impaired spermatogenesis due to the association between the rearranged chromosomes and the sex vesicle. Further studies involving bulls carrying one or more Robertsonian translocations are needed to determine whether this absence of association is a constant feature in cattle.

Animals↗

Synchronously stimulated skeletal muscle graft for left ventricular assistance. Case report.

A 56-year-old man with chronic dilatation of the heart caused by cardiomyopathy (New York Heart Association functional class IV) was selected for cardiomyoplasty. The procedure was divided into two separate stages. In stage I the latissimus dorsi muscle was prepared for progressive stimulation. For this purpose two pacing leads were attached to the muscle, one at the proximal and the other at the distal end, and connected to a pulse-train generator, which was placed subcutaneously. A stimulation protocol was initiated in which the requirements of the latissimus dorsi muscle were periodically increased. In stage II the latissimus dorsi muscle was dissected to make a pedicle graft, which was introduced into the thorax and wrapped around the left ventricle. Pacing electrodes were sewn onto the pedicle graft in the same fashion as in stage I. Two sensing electrodes were fixed to the epicardium of the right ventricle, and the four leads were connected to a double-chambered pacemaker. Twelve days later, the pacemaker was programmed to a bipolar mode, VAT, and the latissimus dorsi muscle graft was synchronously stimulated in a 1:1 assist mode. The patient's status improved to New York Heart Association functional class II. Echocardiographic studies showed better contraction of the posterior wall of the left ventricle, as well as reduction in its diameters. Radioisotopic studies demonstrated a significant improvement in hemodynamic parameters. Cardiomyoplasty seems to be an alternative in those patients with severe cardiac insufficiency caused by cardiac dilation owing to cardiomyopathy.

Cardiac Surgical Procedures↗