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Biomedical subjects

S Cozzolino

Publications and source records attributed to S Cozzolino.

At least 19 recordsLinked to original sources

Follow-up of patients tested for malignant hyperthermia susceptibility.

BACKGROUND AND OBJECTIVE: Malignant hyperthermia is an inherited disorder of skeletal muscle characterized by muscle contracture and hypermetabolic crisis following exposure to halogenated anaesthetics and depolarizing muscle relaxants. We planned this follow-up to get more information about the safety of non-triggering anaesthesia in susceptible patients; the safety of the use of trigger agents in non-susceptible patients and any minor sequelae following the biopsy. METHODS: A questionnaire was sent to 244 patients tested for susceptibility between 1998 and 2004 enquiring about sequelae from the biopsy, subsequent experience with anaesthesia and difficulties encountered because of the investigation. RESULTS: Replies were received from 129 patients. Thirty-four complained about sequelae from the biopsy: 10 reported headache and nausea; 16 experienced pain and a lack of strength in the biopsed leg and 8 found the scar less than satisfactory. Ten patients found it difficult to find a diagnostic centre. Eighteen reported problems and/or delay when they had needed a subsequent anaesthetic. Fourteen patients found the anaesthesiologist reluctant to anaesthetize them and four experienced a delay. Forty-three patients received anaesthesia since their biopsy. Complete medical records were available for 24 anaesthetic exposures in 23 patients. No documented perioperative complications occurred. Only three non-susceptible patients received one trigger agent. CONCLUSIONS: It is safe to use trigger-free anaesthesia in susceptible patients. The difficulties encountered by patients to be anaesthetized and the management of the majority of non-susceptible patients during general anaesthesia show the need of more accurate educational programmes and methods for promoting patient-centred care.

Adolescent↗

Exine micromorphology of Orchidinae (Orchidoideae, Orchidaceae): phylogenetic constraints or ecological influences?

BACKGROUND AND AIMS: Pollen characters have been widely used in defining evolutionary trends in orchids. In recent years, information on pollination biology and phylogenetic patterns within Orchidinae has become available. Hence, the aim of the presented work is to re-evaluate exine micromorphology of Orchidinae in light of recent phylogenetic studies and to test whether pollen micromorphology strictly depends on phylogenetic relationships among species or whether it is influenced by the marked differences in pollination ecology also reported among closely related species. METHODS: Pollen sculpturing of 45 species of Orchidinae and related taxa was investigated using scanning electron microscopy. To cover potential intraspecific variation, several accessions of the same species were examined. KEY RESULTS: Orchidinae show remarkable variation in exine sculpturing, with a different level of variation within species groups. In some genera, such as Serapias (rugulate) and Ophrys (psilate to verrucate), intrageneric uniformity corresponds well to a common pollination strategy and close relationships among species. However, little exine variability (psilate-scabrate and scabrate-rugulate) was also found in the genus Anacamptis in spite of striking differences in floral architecture and pollination strategies. A larger variety of exine conditions was found in genera Dactylorhiza (psilate, psilate-scabrate and reticulate) and Orchis s.s. (psilate, reticulate, perforate-rugulate and baculate) where no unequivocal correspondence can be found to either phylogenetic patterns or pollination strategies. CONCLUSIONS: Changes in pollen characteristics do not consistently reflect shifts in pollination strategy. A unique trend of exine evolution within Orchidinae is difficult to trace. However, the clades comprising Anacamptis, Neotinea, Ophrys and Serapias show psilate to rugulate or scabrate pollen, while that of the clade comprising Chamorchis, Dactylorhiza, Gymnadenia, Orchis s.s., Platanthera, Pseudorchis and Traunsteinera ranges from psilate to reticulate. Comparison of the data with exine micromorphology from members of the tribe Orchidieae and related tribes suggests a possible general trend from reticulate to psilate.

Biological Evolution↗

Inefficient photosynthesis in the Mediterranean orchid Limodorum abortivum is mirrored by specific association to ectomycorrhizal Russulaceae.

Among European Neottieae, Limodorum abortivum is a common Mediterranean orchid. It forms small populations with a patchy distribution in woodlands, and is characterized by much reduced leaves, suggesting a partial mycoheterotrophy. We have investigated both the photosynthetic abilities of L. abortivum adult plants and the diversity of mycorrhizal fungi in Limodorum plants growing in different environments and plant communities (coniferous and broadleaf forests) over a wide geographical and altitudinal range. Despite the presence of photosynthetic pigments, CO2 fixation was found to be insufficient to compensate for respiration in adult plants. Fungal diversity was assessed by morphological and molecular methods in L. abortivum as well as in the related rare species Limodorum trabutianum and Limodorum brulloi. Phylogenetic analyses of the fungal internal transcribed spacer (ITS) sequences, obtained from root samples of about 80 plants, revealed a tendency to associate predominantly with fungal symbionts of the genus Russula. Based on sequence similarities with known species, most root endophytes could be ascribed to the species complex encompassing Russula delica, Russula chloroides, and Russula brevipes. Few sequences clustered in separate groups nested within Russula, a genus of ectomycorrhizal fungi. The morphotypes of ectomycorrhizal root tips of surrounding trees yielded sequences similar or identical to those obtained from L. abortivum. These results demonstrate that Limodorum species with inefficient photosynthesis specifically associate with ectomycorrhizal fungi, and appear to have adopted a nutrition strategy similar to that known from achlorophyllous orchids.

Carbon Dioxide↗

Chloroplast DNA inheritance in the orchid Anacamptis palustris using single-seed polymerase chain reaction.

The modality of chloroplast inheritance in orchids has been investigated only in a few species due to the difficulties associated with the analysis of large progeny numbers from experimental crosses. To test chloroplast DNA inheritance in the orchid Anacamptis palustris, we took advantage of the presence of a highly variable minisatellite repeat located in the tRNA(LEU) intron in the chloroplast genome. Seed progeny obtained from experimental crosses between parental individuals carrying different chloroplast DNA (cpDNA) minisatellite repeat numbers were analyzed using a single-seed polymerase chain reaction (PCR) protocol. All examined seeds displayed the maternal cpDNA haplotypes, indicating that cpDNA inheritance is strictly maternal in this Mediterranean orchid species. No evidence for paternal leakage was found. This finding concurs with results obtained from PCR amplifications of pollen massulae that exclude the presence of chloroplast DNA in the pollen tetrads.

Crosses, Genetic↗

Fine-scale phylogeographical analysis of Mediterranean Anacamptis palustris (Orchidaceae) populations based on chloroplast minisatellite and microsatellite variation.

The phylogeographical history of the rare marsh orchid Anacamptis palustris (Orchidaceae) was reconstructed using highly polymorphic chloroplast minisatellite and microsatellite loci. Allelic variation at chloroplast microsatellite loci was due to length variation in poly(A/T) repeats and was informative on a regional scale, but was not sufficient to unravel relationships among populations on a local geographical scale. The minisatellite locus, however, was found to be highly variable. Nine distinct repeat types were found and variation in repeat number occurred in five repeat types. The distribution of chloroplast haplotypes, combining microsatellite and minisatellite repeat type variation, provided a clear phylogeographical picture on a large geographical scale, whereas length variation in one highly polymorphic minisatellite repeat type provided fine-scale phylogeographical information. Mediterranean populations could be divided into four main lineages, a western European lineage, a northern and central Italian lineage, a well-isolated southern Italian (Apulian) lineage, and an eastern European lineage. Variation at the most variable minisatellite repeat type N revealed 19 alleles and allowed the study of seed-mediated gene flow and an estimation of the ratio of pollen to seed flow among neighbouring populations.

Base Sequence↗

Mutations in the RYR1 gene in Italian patients at risk for malignant hyperthermia: evidence for a cluster of novel mutations in the C-terminal region.

Mutations in the ryanodine receptor type 1 (RYR1) gene are associated with Malignant Hyperthermia (MH) and Central Core Disease (CCD). We report here on the molecular analysis of the RYR1 gene in Italian families referred as potential cases of MH or in patients with CCD or multicore/minicore myopathy. Of a total of 20 individuals with mutations in the RYR1 gene, 14 were part of a group of 47 MH susceptible (MHS) patients, 4 of 34 individuals diagnosed as MH equivocal (MHE), and 2 were patients diagnosed with minicore myopathy and CCD, respectively. Mutations were found to segregate with the MHS or MHE phenotype within the families of the probands. A discordance between phenotype and genotype was observed in a family where a mutation detected in an MHS proband was also found in the father who had been diagnosed MH normal (MHN) at the IVCT. In addition to known mutations, seven novel mutations were found, five of which occurred in exons encoding the C-terminal region of RYR1. These results indicate that the C-terminal region of RYR1 represents an additional hot spot for mutations in patients with MH, similar to what has been reported for patients with CCD.

Amino Acid Sequence↗

Characterization of a minisatellite repeat locus in the chloroplast genome of Orchis palustris (Orchidaceae).

We describe the occurrence of a tandem repeat in the chloroplast genome of the marsh orchid, Orchis palustris. The repeat unit is an AT-rich, 16-bp sequence located in the chloroplast tRNALEU intron. Southern blot analysis confirmed that the O. palustris tRNALEU intron including the minisatellite locus has not been transferred to the nucleus, but is indeed located on the chloroplast genome. The 16-bp repeat unit was found to be present in all O. palustris accessions studied, as well as in the closely related O. laxiflora. Variation in repeat numbers among individuals was found in O. palustris from central and northern Italy; and this was consistently associated with a 13-bp sequence motif preceding the repeat. This motif was absent from O. palustris from southern Italy, Greece, and from O. laxiflora. In these accessions, no variation in repeat numbers was found. Our results suggest that the O. palustris chloroplast minisatellite locus evolved relatively recently, presumably in central Italy, and may represent a valuable marker for population genetic studies.

Base Composition↗

Italian drug information centres: benchmark report.

OBJECTIVE: Pharmacist-operated Drug Information Centres (DICs) in Italy were surveyed to gather current information concerning the status, availability and scope of their services. METHODS: 40 DICs were identified and a questionnaire was mailed to them. Topics covered in the questionnaire included staffing, resources, activities and services. The results were compared with previous survey results to identify trends. RESULTS: Information from 36 DICs was analysed (response rate = 90%). Almost all regions of the country are represented. Pharmacists, part-time or full-time, are the most frequent employees working in the DICs. 91% of DICs participate in the P/T committee, 74% takes part in the technical scientific committee for clinical experimentation and/or to the Medical Ethics committee and 60% participate in the hospital infection committee. 90% of DICs produce newsletters and/or bulletins and are involved in research projects. Regarding the question-answer service, requests are mainly concerned with clinical comparative efficacy, therapeutic use, adverse effects. The most frequent users are physicians (67%) followed by pharmacists (16%) while other professionals use DICs to lesser extent. The most frequently used information sources, include "Goodman and Gilman's the pharmacological basis of therapeutics", Martindale: the Extra Pharmacopeia and Physician's Desk Reference; journals such as the Medical Letter, Adverse Drug Reactions Bulletin and Drug and Therapeutic Bulletin, Micromedex, Medline and IOWA databases. Data collected through the questionnaire and details about organisation and activity of Italian DICs are available in "Eupharma", the web-site of the Italian Society of Hospital Pharmacy (SIFO) at the URL http://www.sifo.it. CONCLUSION: The results parallel the European and American trends. In future communication and collaboration at European level could help to identify best practice models that would result in the standardisation of the activities and in the harmonisation of programs in common areas of interest.

Data Collection↗

Suppression of natural killer cell function and production of soluble ICAM-1: endometrial stroma versus melanoma.

PROBLEM: It has been suggested that specific mechanisms commonly used by different cellular systems to evade immunologic recognition are involved in the development of endometriosis. To gain insight into this aspect, we looked at the relationship between two of these mechanisms in endometrial stroma and the melanoma system for which the ability to create an environment of immune privilege has been well established. METHOD OF STUDY: Media conditioned by endometrial stromal cultures and malignant melanoma A375 were examined to test their effects on peripheral blood mononuclear cell-mediated cytotoxicity directed against K562 target. Moreover, these media were tested for the concentration of the soluble form of intercellular adhesion molecule-1 (sICAM-1), which has been suggested as a marker for spreading potential. RESULTS: Media conditioned by endometrial stromal cultures exerted a significant suppressive effect on cell cytotoxicity when compared with those derived from malignant melanoma Moreover, the constitutive release of sICAM-1 was significantly higher in supernatants from endometrial stromal than in melanoma cells. CONCLUSIONS: These results indicate that two specific properties suggested to be involved in the ability of tumor cells to evade the immune system are more pronounced in the endometrium than in a malignant melanoma. Since the properties evaluated have been previously demonstrated to be even more notable in endometrial samples derived from endometriosis patients, a role of these mechanisms in the development of the disease may be hypothesized.

Cytotoxicity, Immunologic↗

Maternal inheritance of plastids in Encephalartos Lehm. (Zamiaceae, Cycadales).

The mode of inheritance of chloroplast DNA has been determined in Encephalartos by employing a restriction fragment length polymorphism analysis of chloroplast DNA. Artificial F1 hybrids were produced between a female specimen of E. natalensis and a male specimen of E. woodii. The hybridization patterns of all hybrids correspond, in all cases, with that of E. natalensis, and are different from that of E. woodii, thus indicating the maternal inheritance of cpDNA in cycads.

Chloroplasts↗

Dynamic aspects of endometriosis in a mouse model through analysis of implantation and progression.

The aim of this study was to use normal immunocompetent mice to set up a model for endometriosis which allowed to study the dynamic aspects involved in initiation and progression of the disease. Thirty mice were surgically transplanted with autologous endometrium and at 3 weeks showed evidence of endometriosis. Diagnosis of endometriotic lesions was histologically confirmed. Visual inspection using a caliper revealed that, after an initial decrease in size (from 33.44+/-2.33 mm2 to 24.24+/-2.37 mm2 (p<0.01)) detected at 3 weeks after transplantation, there was a significant increase of lesion area from 21.30+/-3.15 mm2 to 43.93+/-6.29 mm2 (p<0.05) in the following 4 weeks. When we compared these results to those obtained in mice which underwent bilateral annessiectomy, we observed that, when bilateral annessiectomy was performed simultaneously to endometrial transplantation, lesion surfaces were similar between mice which were or were not subjected to bilateral ovariectomy. On the other hand, when bilateral annessiectomy was performed at second laparotomy and then evaluated after 4 weeks, differently from what observed in control mice, surface values decreased from 21.24+/-2.29 mm2 to 10.58+/-3.40 mm2 (p<0.01). Finally, progression of lesions in estrogen supplemented mice seems less evident than in control mice since only a slight but not significant increase in size (from 21.32+/-3.32 mm2 to 26.18+/-6.98 mm2, p=0.32) was detected. The results presented herein demonstrate that surgically induced endometrial implants in mice are dynamic lesions and that implantation and progression of endometriosis represent different stages in the ethiopathogenesis of the disease. Moreover, we showed that progression, but not implantation, of ectopic endometrium is dependent upon the functionally and structurally integrity of the ovaries. This is a model of endometriosis established in normal immunocompetent mice, and, consequently, may represent a reliable tool for testing new immunological therapeutical approaches and studying the role of different genes using transgenic mice.

Animals↗

Phytotoxicity to and uptake of flumequine used in intensive aquaculture on the aquatic weed, Lythrum salicaria L.

Phytotoxicity of Flumequine on the aquatic weed Lythrum salicaria L. was determined by two laboratory models: a single concentration test, by which the effects of 100 mg l-1 were evaluated after 10, 20, 30 days and a multiple concentration test, by which the effects of 5000-1000-500-100-50 micrograms l-1 were evaluated after 35-day exposure. 100 mg l-1 are highly toxic and significantly decrease the growth of plants; this effect increases with time. Concentrations between 5000 and 50 micrograms l-1 induced hormesis in plants, by significantly increasing mean number and dimension of leaves and secondary roots. The effect is the highest at 50 micrograms l-1 and decreases with increase in concentration. Both toxic effect and hormesis can be related to plant drug uptake, quite high, in the order of micrograms g-1. The ecological implication of Flumequine contamination in aquatic environments and the possible use of Lythrum salicaria for bioremediation and/or monitoring technique are discussed.

Anti-Infective Agents↗

Molecular analysis of orchid pollinaria and pollinaria-remains found on insects.

Direct observations of pollinator visits to orchids are often difficult and time consuming, especially in orchids with a deceptive pollination system where seed set is typically pollinator-limited. This lack of direct observations greatly inhibits our understanding of orchid-pollinator relationships and especially the degree of pollinator-specificity. Here we describe a molecular approach to the study of orchid-pollinator relationships based on the analysis of DNA recovered from pollinaria found on insects. The insects were collected from nectar-rich plants flowering near natural orchid populations, or taken from museum collections. Sequence analysis of the nuclear ribosomal ITS region allowed the identification of the orchid species or species-group from which the pollinaria originated. Four out of eight orchid-pollinator relationships established with this approach have not been reported previously, which highlights the value of molecular tools for the study of orchid pollination biology.

Animals↗

Phylogeny and evolution of Orchis and allied genera based on ITS DNA variation: morphological gaps and molecular continuity.

Phylogenetic relationships among members of genus Orchis and allied genera Aceras, Anacamptis, Barlia, Dactylorhiza, Gymnadenia, Himantoglossum, Neotinea, Ophrys, Platanthera, and Serapias were inferred from nucleotide sequence variation in the internal transcribed spacer (ITS) regions of nuclear ribosomal DNA. Sequences were subjected to various alignments by changing the gap opening and extension parameters. After a preliminary parsimony analysis, the alignment with the lowest homoplasy indicators was chosen as optimal. The phylogenetic analysis, carried out on the optimal alignment by using Gennaria as an outgroup and a total of 31 taxa, showed that all the genera considered in this study are nested in Orchis despite their distinct morphological features. Genus Orchis is divided into two major clades, each of which includes one or more of the other genera in this study. The resulting phylogenetic hypothesis does not match previous conclusions based on vegetative and floral morphology of the taxa involved but is congruent with isoenzyme, karyological, and chloroplast DNA restriction data. Our results indicate that floral morphology is highly flexible and current generic and infrageneric limits are artificial. Even if some floral characters closely correspond to the molecular data, most are highly homoplastic and thus unsuitable for phylogenetic reconstruction. Various traits pertaining to floral morphology may be interpreted as a result of ecological convergence related to pollinator-mediated selection; such characters can undergo drastic modifications without correspondingly dramatic genetic changes.

Base Sequence↗

Pollination Flow in Hybrid Formation between Orchis morio and Orchis papilionacea (Orchidaceae) in Two Different Habitats.

Two geographically separated natural populations of Orchis xgennarii, a hybrid between Orchis morio and Orchis papilionacea, were examined to establish the parental lineages using nuclear ribosomal DNA and chloroplast DNA length polymorphisms. Results indicate that O. morio more frequently provides the maternal lineage in the population from the volcano Mount Vesuvius (central Campania, Italy) than in the one from Cilento (southern Campania, Italy); in the latter locality maternal genomes are preferentially provided by O. papilionacea. The possible causes of this difference in reproductive behavior are discussed in the light of the pollination biology of parental species and of environmental influences.

Journal Article↗

Mutation screening of the RYR1 gene and identification of two novel mutations in Italian malignant hyperthermia families.

Point mutations in the ryanodine receptor (RYR1) gene are associated with malignant hyperthermia, an autosomal dominant disorder triggered in susceptible people (MHS) by volatile anaesthetics and depolarising skeletal muscle relaxants. To date, 17 missense point mutations have been identified in the human RYR1 gene by screening of the cDNA obtained from muscle biopsies. Here we report single strand conformation polymorphism (SSCP) screening for nine of the most frequent RYR1 mutations using genomic DNA isolated from MHS patients. In addition, the Argl63Cys mutation was analysed by restriction enzyme digestion. We analysed 57 unrelated patients and detected seven of the known RYR1 point mutations. Furthermore, we found a new mutation, Arg2454His, segregating with the MHS phenotype in a large pedigree and a novel amino acid substitution at position 2436 in another patient, indicating a 15.8% frequency of these mutations in Italian patients. A new polymorphic site in intron 16 that causes the substitution of a G at position -7 with a C residue was identified.

Caffeine↗

Significantly improved survival time in pigs with complete liver ischemia treated with a novel bioartificial liver.

Aim of the study was to evaluate treatment efficacy and safety of a scaled-up version of our porcine hepatocytes based BAL system in pigs with complete liver ischemia (LIS). Thirty-one pigs underwent total devascularization of the liver (LIS) by termino-lateral porta-caval shunts and sutures around the bile duct, the common hepatic and gastroduodenal arteries and their accessory branches. The hepato-duodenal ligament was completely transected. Four experimental groups were studied: the first control group (LIS Control, n = 10) received glucose infusion only, the second control group (LIS Plasmapheresis, n = 8) was connected to a centrifugal plasma-separator with a bottle representing the bioreactor volume, the third control group (LIS Empty-BAL, n = 5) received BAL treatment without cells, and the treated group (LIS Cell-BAL, n = 8) was connected for a maximum period of 24 hours to our scaled-up BAL seeded with around 14 billion viable primary porcine hepatocytes. BAL treatment significantly prolonged life in large animals (approximately 35 kg) with complete LIS (Controls, mean +/- SEM: 33.1 +/- 3 h, Cell-BAL: 51.1 +/- 3.4 h; p = 0.001; longest survivor 63 h). In addition, blood ammonia and total bilirubin levels decreased significantly, indicating metabolic activity of porcine hepatocytes in the bioreactor. No significant differences were noticed among the three control groups, indicating that there was no device effect and that the plasmapheresis procedure was well tolerated. No important adverse effects were observed.

Animals↗