Clinical and molecular characterization of two patients with palmoplantar keratoderma-congenital alopecia syndrome type 2.

Palmoplantar keratoderma-congenital alopecia (PPKCA) syndrome is a rare genodermatosis, with two clinically recognizable forms: dominant (Type 1) and recessive (Type 2). Reports of only 18 patients have been published to date, and the molecular basis of the condition is unknown. We describe two cases with PPKCA Type 2 (PPKCA2), comprising a novel patient, originally reported as an example of autosomal ichthyosis follicularis-atrichia-photophobia syndrome, and the 6-year follow-up of a previously published case. Extensive molecular studies of both patients excluded mutations in all the known genes associated with PPK and partially overlapping syndromes. The striking similarities between these two patients confirm PPKCA2 as a discrete genodermatosis, of which the main features are congenital and universal alopecia, diffuse keratosis pilaris, facial erythema, and a specific PPK with predominant involvement of the fingertips and borders of the hands and feet, with evolution of sclerodactyly, contractures and constrictions. Clinical follow-up of these patients has demonstrated progressive worsening of the hand involvement and attenuation of facial erythema.

Thricho-rhino-phalangeal syndrome and severe osteoporosis: a rare association or a feature? An effective therapeutic approach with biphosphonates.

Trichorhinophalangeal syndrome (TRPS) is a rare, autosomal dominant malformation syndrome characterized by hair, craniofacial and skeletal abnormalities, skin laxity, deformation of phalanges and anomalies of pelvis, femurs, and tibias. Three subtypes have been described: TRPS I, caused by mutations in TRPS1 gene on chromosome 8; TRPS II, a microdeletion syndrome affecting the TRPS1 and EXT1 genes; and TRPS III, a form with severe brachydactyly, due to short metacarpals, and severe short stature, but without exostoses. We present the case of a 7-year-old boy, affected by TRPS with a severe osteoporosis and several spontaneous bone fractures, an association described only once in the literature, successfully treated with biphosphonates. Bone mineral density (BMD) at dual-energy X-ray Absorptiometry (DXA) was of 0.331 g/cm(2) at lumbar spine with. He had four spontaneous femoral fractures in a year, and for this reason he was been operated for positioning intramedullary osteosynthesis and orthopedic supports. Due to the severity of the clinical and radiological pattern it was established, after approval of the Ethical Committee, to begin off-label therapy with infusions of neridronate at a dose of 2 mg/kg IV every 3 months. The treatment was, in this patient, effective both in terms of clinical (absence of new fractures) and mineralomethric (+45% BMD ath the lumbar level). We therefore suggest that treatment with biphosponates can be taken in account as a possible therapeutic option in case of bone fragility in patients with TRPSI.

Syndromic non-compaction of the left ventricle: associated chromosomal anomalies.

Non-compaction of the left ventricle (NCLV) is a cardiomyopathy characterized by prominent left ventricular trabeculae and deep intertrabecular recesses. Associated extracardiac anomalies occur in 14-66% of patients of different series, while chromosomal anomalies were reported in sporadic cases. We investigated the prevalence of chromosomal imbalances in 25 syndromic patients with NCLV, using standard cytogenetic, subtelomeric fluorescent in situ hybridization, and array-comparative genomic hybridization (CGH) analyses. Standard chromosome analysis disclosed an abnormality in three (12%) patients, including a 45,X/46,XX mosaic, a 45,X/46,X,i(Y)(p11) mosaic, and a de novo Robertsonian 13;14 translocation in a child affected by hypomelanosis of Ito. Cryptic chromosome anomalies were found in six (24%) cases, including 1p36 deletion in two patients, 7p14.3p14.1 deletion, 18p subtelomeric deletion, 22q11.2 deletion associated with velo-cardio-facial syndrome, and distal 22q11.2 deletion, each in one case. These results recommend accurate clinical evaluation of patients with NCLV, and suggest that chromosome anomalies occur in about one third of syndromic NCLV individuals, without metabolic/neuromuscular disorder. Array-CGH analysis should be included in the diagnostic protocol of these patients, because different submicroscopic imbalances are causally associated with this disorder and can pinpoint candidate genes for this cardiomyopathy.

Postharvest decay control of citrus fruit by preharvest pyrimethanil spray.

Preharvest infections or conidia load on fruit surface by Penicillium digitatum, P. italicum, Alternaria citri and other filamentous fungi can cause important postharvest losses of citrus fruit. Reduction in pruning frequency occurred in the last decade together with un-picked yield that eventually rots on the trees have increased the risk of postharvest decay especially when environmental conditions at picking time are favourable to pathogens' development. Sanitation procedures in the packinghouses, alternate use of postharvest fungicides with different modes of action, along with fungicide application before harvest could be an effective approach to minimize postharvest decay in citrus fruit. The present study investigated the effectiveness of a preharvest treatment with pyrimethanil (PYR), a broad spectrum fungicide, recently registered in different citrus-producing countries for postharvest treatments of citrus fruit and widely used worldwide as a preharvest treatment to control various diseases in different crops. PYR (750 mg/L) was sprayed by a hand-back sprayer at run-off on 'Fremont' mandarins. The day after the treatment, half of the trees were sprayed with a 10(4) conidial suspension of P. digitatum at run-off. Fruit were harvested following 2 or 4 weeks from treatments. Sound or either wounded 2-mm-deep and 2-mm-wide or superficial wound-scratched fruit were stored at 20 degrees C and 90% RH and inspected for decay after 1, 2 or 3 weeks of storage. In fruit harvested after 2 weeks from field treatment, PYR remarkably reduced decay development during two weeks of storage in sound fruit and in wound-scratched fruit and was fairly effective even after 4 weeks from treatment, but was ineffective in fruit wounded 2 mm deep and 2 mm wide. PYR was also effective in reducing preharvest decay incited by P. digitatum, P. italicum and Botrytis cinerea, but not by other pathogens. Results show that preharvest treatment with PYR could be a feasible approach to reduce postharvest chemical control of decay of citrus fruit.

Residue levels, persistence and effectiveness of imazalil against a resistant strain of penicillium digitatum when applied in combination with heat and sodium bicarbonate.

Green and blue molds, respectively caused by Penicillium digitatum Sacc., and P. italicum Wehmer, are the most important postharvest diseases of citrus fruit Postharvest management of these pathogens is mainly based on the application of thiabendazole (TBZ) or imazalil (IMZ) fungicides. However, their intensive and prolonged use has led to the selection of TBZ- IMZ-resistant strains of these pathogens and to a reduction of TBZ and IMZ effectiveness to control postharvest decay. However, while TBZ may become completely ineffective against TBZ-resistant strains of P. digitatum, reduction of IMZ efficacy is only partial, and an effective control of decay can still be achieved by increasing its concentration, heating the treatment-solution and/or combining IMZ with sodium bicarbonate (SBC) or other food additives or natural salts. In this study, 'Desiderio' and 'Nova' mandarins were inoculated with spores of a sensitive strain of P. digitatum to IMZ and TBZ (PDs) or with a strain of P. digitatum with double resistance to both fungicides (PDr) and immersed in IMZ or TBZ emulsions at increasing concentrations up to 1000 mg/L or in IMZ (25, 200 or 400 mg/L), SBC (0.5, 1 or 2%) or IMZ + SBC emulsions either at 20 or 40 degrees C. IMZ was superior to TBZ to control decay of 'Desiderio' mandarins incited by PDs and was also effective to control decay in fruit inoculated with PDr, while TBZ even at the highest rate was completely ineffective. In 'Desiderio' mandarins inoculated with PDs, a complete control of decay was achieved with 25 mg/L IMZ but in fruit inoculated with PDr, 25 mg/L IMZ were ineffective to control decay despite in combination with SBC at 2% a synergistic effect was detected. In contrast, a good control of decay was achieved with 400 mg/L IMZ. In 'Nova' mandarins after 1 week of incubation at 20 degrees C decay incidence in fruit dipped in 400 mg/L at 20 degrees C or 200 mg/L IMZ at 40 degrees C was almost completely inhibited, while the addition of SBC at 0.5, 1 or 2% did not improve treatments performance in fruit inoculated with PDs. However, when 'Nova' mandarins were inoculated with PDr, SBC showed a modest but significant control of decay and in combination with IMZ either at 400 mg/L and 20 degrees C or 200 mg/L and 40 degrees C, significantly improved decay control. SBC did not affect IMZ residue load in 'Valencia' oranges, whereas dipping the fruit in 400 mg/L IMZ at 20 degrees C produced similar IMZ residue load as dips at 200 mg/L IMZ at 40 degrees C. In all cases, residue levels of IMZ never exceeded 2 mg/kg, which is about 40% of the maximum residue limits (MRLs) allowed in European countries. Thus, despite the selection of IMZ-resistant strains of P. digitatum, IMZ continues to be highly effective to control green mold of citrus fruit at concentrations leaving on fruit surface residue levels below the MRLs.

Mild ring 17 syndrome shares common phenotypic features irrespective of the chromosomal breakpoints location.

Ring 17 syndrome is a rare disorder with clinical features influenced by the presence or deletion of the Miller-Dieker critical region (MDCR). Presence of the MDCR is associated with a mild phenotype, including growth delay (GD), mental retardation (MR), seizures, cafè au lait skin (CALS) spots and minor facial dysmorphisms. Previous studies have been mainly focused on this locus providing poor information about the role of other genes located on the p- and q-arms. Here, we used bacterial artificial chromosome (BAC)/P1 artificial chromosome (PAC) and fosmid clones as fluorescence in situ hybridization (FISH) probes to perform a cyto-molecular analysis of a ring 17 case and found that the breakpoints were close to the telomeric ends. METRNL is the sole gene located on the q-arm terminal end, whereas two open reading frames and the RPH3AL gene are located on the terminal p-arm. To detect possibly unrevealed small deletions involving the transcription units, we used subcloned FISH probes obtained by long-range polymerase chain reaction (PCR), which showed that the investigated regions were preserved. Comparing our findings with other reports, it emerges that different breakpoints, involving (or not) large genomic deletions, present overlapping clinical aspects. In conclusion, our data suggest that a mechanism based on gene expression control besides haploinsufficiency should be considered to explain the common phenotypic features found in the mild ring 17 syndrome.

Influence of post-harvest treatments with fludioxonil and soy lecithin co-application in controlling blue and grey mould and fludioxonil residues in Coscia pears.

The residue levels of fludioxonil (FLU) were determined in Coscia pear following a 1-, 2- or 4-min dip in an aqueous mixture of FLU containing 300 or 100 mg l(-1) (active ingredient, a.i.) at 20 and 50 degrees C, respectively, with or without 2% soy lecithin. The efficacy of heat treatment with water and FLU mixtures was investigated on artificially inoculated pears for the control of post-harvest decay caused by blue (Penicillium expansum Link) and grey (Botrytis cinerea Pers. ex Fr.) mould. Treatment with 300 mg l(-1) FLU at 20 degrees C increased residues significantly when treatment time rose from 1 to 2 min; no further increase was recorded when dip time raised from 2 to 4 min. FLU residue rates were unaffected by treatment time when 300 mg l(-1) a.i. was applied in combination with lecithin at 20 degrees C. While treatment with 100 mg l(-1) a.i. at 50 degrees C for 1 and 2 min resulted in similar residue levels, significantly higher residues were detected when dip time increased from 1 to 4 min. Co-application of lecithin significantly decreased FLU residues with respect to fruit treated with FLU alone. Treatments with FLU at 20 or 50 degrees C effectively controlled decay over 10 days of incubation. While co-application of lecithin did not affect the efficacy of FLU at 300 mg l(-1)and 20 degrees C, treatment efficacy decreased when lecithin was applied in combination with 100 mg l(-1) FLU and 50 degrees C for 4 min and to a greater extent when dip time was 1-2 min.

Masked complex chromosome rearrangement in a child thought to have del(8qter) as the sole cytogenetic abnormality.

Cytogenetic analyses of constitutional diseases have disclosed several chromosomal rearrangements. At the molecular level, these rearrangements often result in the breakage of genes or alteration of genome architecture. Fluorescence in situ hybridization (FISH) and molecular investigations of a patient showing hypotonia and dysmorphic traits revealed a masked complex chromosome abnormality previously detected by G-banding as a simple 8qter deletion. To characterize the genetic rearrangements panels of bacterial artificial chromosomes (BACs) covering 8q24.22-->qter were constructed, and short tandem repeats (STRs) were used to refine the localization of the breakpoints and to assess the parental origin of the defect. Chromosome 8 displayed the breakpoint at 8q24.22 and an unexpected distal breakpoint at 8q24.23 resulting in unbalanced translocation of a small 8q genomic region on the chromosome 16qter. The study of the 16qter region revealed that the 16q subtelomere was retained and the translocated material of distal 8q was juxtaposed. Moreover, molecular analyses showed that part of the translocated 8qter segment on der(16) was partially duplicated, inverted and that the rearrangement arose in the paternal meiosis. These findings emphasize the complexity of some only apparently simple chromosomal rearrangements and suggest a subtelomeric FISH approach to enhance diagnostic care when a cytogenetic terminal deletion is found.

Dopamine D2 receptor gene polymorphisms and response to cabergoline therapy in patients with prolactin-secreting pituitary adenomas.

Dopamine-agonist cabergoline (CB) reduces prolactin (PRL) secretion and tumor size in 80% of patients with prolactin-secreting adenomas (PRL-omas) by binding type 2 dopamine receptor (DRD2). The mechanisms responsible for resistance to CB remain largely unknown. To assess the association of DRD2 with sensitivity to CB, TaqI-A1/A2, TaqI-B1/B2, HphI-G/T and NcoI-C/T genotypes were determined in a cross-sectional retrospective study, including 203 patients with PRL-oma. DRD2 alleles frequencies did not differ between patients and 212 healthy subjects. Conversely, NcoI-T allele frequency was higher in resistant rather than responsive patients, considering both PRL normalization (56.6 vs 45.3%, P=0.038) and tumor shrinkage (70.4 vs 41.4%, P=0.006). Finally, [TaqI A1-/TaqI B1-/HphI T-/NcoI T-] haplotype was found in 34.5% of patients normalizing PRL with < or =3 mg/week of CB vs 11.3% of resistants (P=0.021). In conclusion, resistance to CB was associated with DRD2 NcoI-T+ allele, consistent with evidence suggesting that this variant may lead to reduction and instability of DRD2 mRNA or protein.

Diagnosis of atypical CF: a case-report to reflect.

Non-classic Cystic Fibrosis (CF) still represents a difficult entity to diagnose. We present a case of two sisters affected by mild pulmonary symptoms started at puberty, carriers of the F508del mutation associated with the T5TG13 combination. We discuss the clinical utility of TG repeat testing in individuals carrying the T5 variant. Furthermore, this case-report leads to reflect on the natural history of CF and the correct management of its atypical forms.

Comparative analysis of polyphenolic profiles and antioxidant and antimicrobial activities of tunisian pome fruit pulp and peel aqueous acetone extracts.

Pome trees, apple, pear, and quince, are classified into the subfamily Pomoideae, belonging to the Rosaceae family. Their autumnal fruits are consumed worldwide in different forms, that is, fresh or transformed into jams, jelly, juices, etc. Their well-established beneficial properties to human health were found mainly related to their phenolic content. Pulp and peel aqueous acetone extracts obtained from Tunisian fruits at commercial maturity were comparatively evaluated for their phenolic profiles and antioxidant and antimicrobial potentials. The phenolic compounds present in the extracts were identified and quantified using RP-HPLC-DAD and ESI-MS techniques. Significant differences in the chromatographic profiles among these fruits, as well as between pulp and peel extracts of each fruit, were observed. Quince, followed by 'Red Delicious', peel extracts showed the highest phenolic content (160.33 and 110.90 mg/100 g of fresh weight). The stronger inhibitory effect on DPPH radicals corresponded to those obtained from peel materials. A comparative analysis of the antimicrobial potential against a range of microorganism strains was also carried out. Staphylococcus aureus, Pseudomonas aeruginosa, and Bacillus cereus were the most sensitive to the active extracts. Among the examined phenolic extracts, 'Red Delicious' and quince peels showed the highest effects for inhibiting bacteria growth. Minimum inhibitory and bactericide concentrations ranged from 10(2) to 10(4) microg of polyphenol/mL. Red skin apple and quince peels could be of great interest as important antioxidant and antimicrobial polyphenol sources.

In vitro interaction between ochratoxin A and different strains of Saccharomyces cerevisiae and Kloeckera apiculata.

The interaction of ochratoxin A (OTA) and 20 yeast strains of Saccharomyces cerevisiae and Kloeckera apiculata during alcoholic fermentation was studied. Levels of OTA were determined in the fermentation liquid and in the yeast cells solid using a high-performance liquid chromatography system with a fluorescence detector. Yeast cells do not adsorb OTA, and for all yeasts, OTA levels did not affect the alcoholic fermentation. Some yeast strains reduced levels of OTA, whereas other strains did not show any effect demonstrating that OTA level reduction is not a genus species characteristic but a strain trait.

Effects of chronic retinoid administration on pituitary function.

It has been reported that retinoids may affect hypothalamic-pituitary-thyroid axis, causing central hypothyroidism. In the present study, we evaluated pituitary function in 11 male psoriatic patients at baseline and after 1 and 3 months of treatment with acitretin (all-trans retinoic acid, 35 mg/day). Serum LH, FSH, testosterone, cortisol, GH and IGF-I levels were not affected by the treatment. By contrast, we observed a significant decrease in TSH levels (from 0.92 +/- 0.3 to 0.80 +/- 0.3 mU/I, p < 0.05) at 1 month, that reverted to baseline after 3 months. No change in free T4 (FT4) levels occurred, while free T3 (FT3) levels were reduced at 1 and 3 months (from 6.7 +/- 0.5 to 6.2 +/- 0.3 and 6.1 +/- 0.6 pmol/l; p < 0.05, respectively). Moreover, acitretin treatment induced a significant reduction of PRL levels after 3 months (from 182 +/- 70 to 150 +/- 56 mU/l, p < 0.05). During treatment, no change in TSH and PRL response either to TRH or dopamine infusion was observed. In conclusion, we demonstrated that treatment with low dose of acitretin induced a series of hormonal modifications that, in addition to a mild and transient reduction of TSH levels, included a persistent reduction of FT3, probably due to changes in thyroid hormone metabolism, and a decrease in PRL levels.

Residues of azoxystrobin, fenhexamid and pyrimethanil in strawberry following field treatments and the effect of domestic washing.

Residues of the pesticides azoxystrobin, fenhexamid and pyrimethanil were determined in strawberry after field treatment. The effect of 'home' washing with tap water and a commercially available vegetable detergent on residue levels was also studied. After treatment, azoxystrobin and pyrimethanil residues on strawberry were on average 0.55 and 2.98 mg kg(-1), respectively, values below the maximum residue level (MRL) fixed by the European Union (2.0 and 5 mg kg(-1), respectively), while fenhexamid residues were on average 2.99 mg kg(-1), which is very close to the MRL of 3.0 mg kg(-1), but some samples were over the MRL. Thereafter, all residues declined, with a half-life of about 8 days (azoxystrobin and fenhexamid) and 4.8 days (pyrimethanil). Washing the fruit with tap water reduced the residues of azoxystrobin and fenhexamid but did not affect pyrimethanil residues. Finally, when fruits were washed with a commercial detergent, greater amounts were removed (about 45% of azoxystrobin and pyrimethanil and 60% of fenhexamid).

Triazole fungicide degradation in peaches in the field and in model systems.

The degradation of five triazole fungicides (cyproconazole, hexaconazole, penconazole, propiconazole, tebuconazole) in peaches was evaluated in field studies to establish whether at the preharvest interval the residue levels were below the legal limit established in Italy. All fungicides, except propiconazole, showed higher residues than the legal limits because of their high stability. In fact, some did not decrease at all (cyproconazole, penconazole, tebuconazole), while others decreased only in part. The increasing weight of the fruit during growth was the main factor leading to an apparent decrease in pesticide levels on a weight for weight basis. The trials on model systems showed that co-distillation and sunlight photodegradation were the main mechanisms leading to a decrease in triazole levels. Although these fungicides are systemic, they did not enter the fruit and all residues were only present in the peel.

Spectrum of clinical variability in familial deletion 22q11.2: from full manifestation to extremely mild clinical anomalies.

The 22q11.2 deletion (del22q11.2) syndrome is a genetic condition with wide interfamilial and intrafamilial variability in clinical expression. The aim of the present study was to review the prevalence of parental transmission in our series of patients with del22q11.2, and to analyse clinical findings of the affected parents. Parental transmission of del22q11.2 in our series was 17.2% (15/87), with a preferential maternal transmission (10/15). One or more major features of del22q11.2 were found in all deleted parents, but one of the mothers showed extremely mild clinical anomalies. The present data demonstrate that it should be current policy to test both parents of patients with del22q11.2, irrespective of the parental phenotype, in view of the fact that extremely mild clinical features can be detected in parents of deleted patients. This would provide accurate genetic counselling to del22q11.2 families, as relatively asymptomatic parents must be advised of the 50% risk of transmitting the deletion in a subsequent pregnancy. Various genetic and non-genetic factors, including modifier genes at separate loci, mosaicism, unstable mutations, allelic variations at the haploid locus, chance and environmental interaction, can be hypothesized to be involved in variable clinical expression, even in the same family.

The effect of simulated rain on folpet and mancozeb residues on grapes and on vine leaves.

Artificial rainfalls were used to determine the effect of the amount of the rainfall and the time interval between pesticide application and rainfall event, on folpet and mancozeb residues on grapes and vine leaves. Forty-five mm of rain were administered to the vineyard in different amounts (45; 30+15; 15+15+15 mm). Folpet showed good rainfastness on the grapes and on the leaves. A modest decrease was observed only in the experiments that had received 45 mm of rain at one go. Mancozeb showed a lower rainfastness, since a portion of the deposit was easily washed off also by a modest rainfall. The percentage of this portion was higher in the grapes (38%) than in the leaves (20%). The data obtained in these experiments show that, in the case of folpet, it is not necessary to repeat the treatment when it rains the day after, while it is recommendable to repeat it in the case of mancozeb.

Fenhexamid residues in grapes and wine.

The behaviour of the fungicide, fenhexamid, on grapes, and during wine-making, as well as its effect on the microflora of alcoholic and malolactic fermentation has been studied. After treatment, the residue on grapes decreased rapidly to one-third of the initial level after the first week, while it remained constant during the following two weeks. At harvest, in the wine obtained by vinification without skins, the fungicide residue decreased on average by 49%, while in the wine obtained by vinification with skins, the decrease was on average 62%. The presence of this fungicide on grapes and in the wine did not affect alcoholic and malolactic fermentation, nor did fermentation cause any degradation of the fungicide. A simple and rapid gas chromatograhic method (GC-NPD) for the determination of fenhexamid residues in grapes, must and wine is described.