Role of genetic investigation in the diagnosis of short stature in a cohort of Italian children.

BACKGROUND: Short stature (SS) is defined as height more than 2 standard deviations below the mean for age and sex. Hypothyroidism, celiac disease, growth hormone deficiency, hormonal abnormalities, and genetic conditions are among its causes. A wide range of conditions often due to largely unknown genetic variants can elude conventional diagnostic workup. AIM: We used next-generation sequencing (NGS) to better understand the etiology of SS in a cohort of Italian children. PATIENTS AND METHODS: The study sample was 125 children with SS of unknown origin referred to our Institute between 2015 and 2021. All had undergone complete auxological and hormonal investigations to exclude common causes of SS. Genetic analysis was performed using a NGS panel of 104 genes. Clinical data were reviewed to clarify the pathogenicity of the variants detected. RESULTS: In this cohort, 43 potentially causing variants were identified in 38 children. A syndromic genetic condition was diagnosed in 7: Noonan syndrome in 3, Leri-Weill syndrome in 3, and hypochondroplasia in 1. Moreover, 8 benign variants and other 37 like benign variants were found. In 88 children, 179 variants of uncertain significance (VUS) were identified. No variant was found in 16 children. CONCLUSION: Genetic analysis is a useful tool in the diagnostic workup of patients with SS, in adapting management and treatment, and in identifying syndromes with mild atypical clinical features. The role of VUS should not be underestimated, particularly when multiple VUS with possible mutual worsening effects are present in the same child.

Coeliac Disease High-Risk Human Leukocyte Antigen Alleles in a South African Type One Diabetic Population.

BACKGROUND: Coeliac disease (CD) is an autoimmune disorder strongly associated with Human Leukocyte Antigen (HLA) genes DQ2 and DQ8. These alleles are also associated with other autoimmune diseases including type 1 dia-betes (T1D) and these patients are consequently at a higher risk of developing CD. We examined the frequency of high-risk HLA alleles in a cohort of South African T1D pediatric patients. METHODS: A total of 22 pediatric participants were recruited at an endocrinology clinic in a public sector hospital in Johannesburg, South Africa. Clinical details were collected, and all patients were HLA typed at the DQ loci. RESULTS: Of our T1D patients, 63.6% had at least 1 high-risk HLA type alleles associated with CD. Of the patients with high-risk alleles, DQ2 was found in the majority which is consistent with CD literature. CONCLUSIONS: This study provides preliminary insight into the frequency of the HLA-DQ types associated with CD in a South African population. The high prevalence of high-risk alleles in our T1D population motivates continual monitoring and further investigations into CD in South Africa.

Production and characterization of 111Ag radioisotope for medical use in a TRIGA Mark II nuclear research reactor.

Radio Pharmaceutical Therapy (RPT) comes forth as a promising technique to treat a wide range of tumors while ensuring low collateral damage to nearby healthy tissues. This kind of cancer therapy exploits the radiation following the decay of a specific radionuclide to deliver a lethal dose to tumor tissues. In the framework of the ISOLPHARM project of INFN, 111Ag was recently proposed as a promising core of a therapeutic radiopharmaceutical. In this paper, the production of 111Ag via neutron activation of 110Pd-enriched samples inside a TRIGA Mark II nuclear research reactor is studied. The radioisotope production is modeled using two different Monte Carlo codes (MCNPX and PHITS) and a stand-alone inventory calculation code FISPACT-II, with different cross section data libraries. The whole process is simulated starting from an MCNP6-based reactor model producing the neutron spectrum and flux in the selected irradiation facility. Moreover, a cost-effective, robust and easy-to-use spectroscopic system, based on a Lanthanum Bromo-Chloride (LBC) inorganic scintillator, is designed and characterized, with the aim of using it, in the future, for the quality control of the ISOLPHARM irradiated targets at the SPES facility of the Legnaro National Laboratories of INFN. natPd and 110Pd-enriched samples are irradiated in the reactor main irradiation facility and spectroscopically characterized using the LBC-based setup and a multiple-fit analysis procedure. Experimental results are compared with theoretical predictions of the developed models, showing that inaccuracies in the available cross section libraries prevent an accurate reproduction of the generated radioisotope activities. Nevertheless, models are normalized to our experimental data allowing for a reliable planning of the 111Ag production in a TRIGA Mark II reactor.

3D printable fully biomass-based composite using poly(furfuryl alcohol) as binder and cellulose as a filler.

Nowadays, composite materials are widely used in different sectors owing to their improved mechanical and functional properties compared to bulk materials and efficient manufacturing processes. Nevertheless, the majority of these materials are still petroleum-based, which is incompatible with the recent environmental awareness. As a result, in the current study, a fully biomass-based composite material was produced employing poly(furfuryl alcohol) (PFA) as a bio-based matrix coupled with cellulose powder as fillers and processing aid agent. The addition of cellulose powder increased the viscosity of the uncured composite paste and conferred it a shear-thinning thixotropic making it suitable for 3D printing using the liquid deposition modeling technique (LDM). After curing, the combination of these raw materials yields a renewable and cost-effective composite for additive manufacturing by the LDM technique with high interlayer and interfilament adhesion, good mechanical performances, and adequate shape fidelity.

Effect of different carboxylic acids in cyclodextrin functionalization of cellulose nanocrystals for prolonged release of carvacrol.

Current investigations deal with new surface functionalization strategy of nanocrystalline cellulose-based substrates to impart active molecule release properties. In this study, cellulose nanocrystals (CNC) were surface-functionalized with ß-cyclodextrin (ß-CD) using succinic acid (SA) and fumaric acid (FA) as bridging agents. The main objective of this surface modification performed only in aqueous media was to obtain new active materials able to release antibacterial molecules over a prolonged period of time. The reactions were conducted by immersing the CNC film into a solution composed of ß-CD, SA and FA, leading to CNC grafting. The materials were characterized by infrared spectroscopy (FT-IR), Quartz crystal microbalance-dissipation (QCM-D), AFM and phenolphthalein (PhP) was used to determine the efficiency of CNC grafting with ß-CD. The results indicated that ß-CD was successfully attached to the CNC backbone through the formation of ester bonds. Furthermore, carvacrol was entrapped by the attached ß-CD and a prolonged release was confirmed. In particular, CNC grafted to ß-CD in the presence of FA was selected as the best solution. The antibacterial activity and the controlled release were studied for this sample. Considerably longer bacterial activity against B. subtilis was observed for CNC grafted to ß-CD compared to CNC and CNC-FA, confirming the promising impact of the present strategy.

[Rectal cancer margin]. / Marge du cancer du rectum.

Treatment of rectal cancer has dramatically improved over the last twenty years. During previous years surgeons and pathologists have focused mainly on distal longitudinal margin as the main parameter for anal conservation. New knowledge on the prognostic role of circumferential margin and its influence on locoregional recurrence rate and patients' survival has led to a more standardized surgery and the definition of the total mesorectal excision concept. As a consequence the rates of positive circumferential margins and locoregional recurrence have decreased in population studies as well as in prospective randomized trials.

A technique of mRNA extraction and labeling from circulating lymphocytes of children treated with growth hormone replacement therapy for microarray analysis.

GH replacement therapy exhibits a wide spectrum of response in terms of growth. Nevertheless, standardized doses are still given in clinical practice. In order to optimize the therapy, it is necessary to identify its markers of responsiveness. Given the presence of GH receptors in the circulating lymphocytes, accessible by means of a simple blood withdrawal, blood becomes the tissue of choice as a source of RNA for in vivo gene expression analysis. Hence, the purpose of the present paper is to develop a method of preparation of RNA from lymphocytes suitable for microarray analysis, focusing on the reduction of the blood volume withdrawal in order to perform the analysis on pediatric subjects. After lymphocyte isolation and total RNA extraction from 6 ml of blood, we carried out an amplification procedure preserving the relative abundance of each transcript. Thereafter, we hybridized the labeled amplified RNA on an oligo chip (Human 30K A, MWGBiotech), but the unsuccessful detection of a good signal to noise ratio indicates that labeled RNA is still insufficient. Therefore, we suggest performing pools of total RNA from different subjects with similar responsiveness to the therapy. It can be speculated that, upon comparison of the obtained data with those derived from pools of controls properly responding to the therapy, specific hallmarks of the condition of low responsiveness, devoid of inter-individual variability, will be evidenced.

Novel thermoplastic materials based on the outer-shell oxypropylation of corn starch granules.

The study of the plasticization of corn-starch granules through the bulk oxypropylation of their outer shell produced a novel biphasic material consisting of a low glass transition temperature component surrounding the granules' inner core, which could be hot pressed to form films of the granules dispersed into a thermoplastic matrix. The success and extent of these chemical modifications and the properties of the ensuing composites were assessed by Fourier transform infrared spectroscopy, X-ray diffraction, scanning electron microscopy, differential scanning calorimetry, thermogravimetric analysis, and contact angle measurements. This approach has the additional advantage of being simple and "green".

Genetic analysis carried out on blood-spots of phenylalanine hydroxylase-deficient newborns detected by northeastern Italian neonatal screening.

The aim of this work was to perform genetic analysis on 18 different blood-spot samples collected from neonates detected as hyperphenylalaninemic by Northeastern Italian screening program. DNA was extracted from blood-spots. Exons/introns of PAH gene were amplified by polymerase chain reaction (PCR), and PCR products were purified and sequenced with both forward and reverse primers. The most frequent mutations were IVS12nt1g>a (16.7%) and R408W, P281L and L48S (all together 11.1%). As expected, compound heterozygosity was the usual finding; homozygosity was found only in two patients with R158Q and IVS2nt5g>c mutations. The V230I mutation was reported for the first time in Italy. We found six previously described polymorphisms (V245V, IVS4nt47c>t, IVS2nt19t>c, IVS3nt-22c>t, IVS5nt-54a>g, and E280>Q280). To our knowledge, four genotypes were not previously described: R158Q/V230I present in one patient with classical PKU; and L48S/R408Q, A403V/IVS2nt-13t>g, and G272X/V230I present in patients showing HPA phenotype. Most of the mutations were located in the exons 12 and 7 and in exon/intron 2 (83.3% detection of total mutations in PKU or HPA patients of Northeastern Italy). From a practical viewpoint, the genetic analysis of blood-spots collected on Guthrie cards for neonatal screening for PKU could be a simple method to establish the genotype of neonates. Consequently, the genotype/phenotype correlation could lead to a more accurate diagnosis and prognosis for families.

Modification of cellulose fibres with organosilanes: Under what conditions does coupling occur?

Different cellulose substrates and organosolv lignin were treated heterogeneously with organic solutions of trialkoxysilanes bearing variable function on the forth substituent. It was shown unambiguously for the first time that Si-OR does not react with the hydroxy groups of cellulose even at high temperature, whereas it condenses with the phenolic OH of lignin. The addition of moisture to these systems induces the partial hydrolysis of the siloxane moieties and the ensuing silanol groups can then react with the cellulose OH, but only at high temperature. Using the latter systems and a siloxane bearing a polymerizable function, it was possible to attach poly(methylmethacrylate) chains to the surface of cellulose fibres through a two-step procedure.

Oxypropylation of cork and the use of the ensuing polyols in polyurethane formulations.

Cork particles, recovered as byproducts of the processing of this natural material, were oxypropylated under pressure and relatively high temperature in the presence of KOH as catalyst. Various parameters were explored in order to assess the most suitable conditions, which led to the almost complete conversion of the solid cork into a viscous polyol. This product was a mixture of oxypropylated cork macromolecules and propylene oxide oligomers, which were thoroughly characterized. The use of these polyols as macromonomers in the synthesis of polyurethane foams gave promising results, thus showing that it should be possible to exploit the residues of this important renewable resource to manufacture original materials.

Unrelated allogeneic bone marrow donation: short- and long-term follow-up of 103 consecutive volunteer donors.

Between 1992 and 1999, 105 unrelated allogeneic bone marrow collections from 103 volunteer donors (65 males and 38 females; median age 33 years) were carried out in three northern Italian centers (Verona, Bolzano and Padova) affiliated with the Italian Bone Marrow Donor Registry (IBMDR). The average volume of BM collected was equivalent in both genders (1143.1 ml for males and 1054.2 ml for females; P = 0.1), although the average volume collected for unit of body weight and the average post-collection blood volume depletion was higher in females (respectively 17.1 ml/kg and 14.2% in females, 14.8 ml/kg and 12% in males; P= 0.01 and 0.03). There was no statistically significant difference between males and females in the total number of nucleated cells collected. We did not record any acute life-threatening event during or after the bone marrow collections. The most frequent complaint was pain at the collection site (77%) followed by the onset of fatigue (38%) and nausea and vomiting (25%); all of these were short-term problems. Hospitalization was short (average 20.2 h) and donors started their normal daily activities after an average of 5.4 days. We also monitored Hb, serum ferritin levels, WBC and platelet counts in the post-collection period (average follow-up 40.1 months). All donors signed a written informed consent for a further bone marrow collection, if needed. Our findings confirm the short- and long-term safety of allogeneic bone marrow collection in volunteer donors.

Cork suberin as a new source of chemicals. 1. Isolation and chemical characterization of its composition.

Extractive-free cork from Quercus suber L. was submitted to a solvolysis treatment with methanolic NaOH which yielded 37% (o.d. cork) of suberin. This mixture of compounds was thoroughly characterized by FTIR, 1H- and 13C-NMR, gas chromatography coupled with mass spectrometric (GC-MS) analysis, vapour pressure osmometry (VPO), mass spectrography (MS) and gel permeation chromatography (GPC). After derivatization, the main components of the volatile fraction, representing less than half of the total, were found to be omega-hydroxymonocarboxylates, alpha, omega-dicarboxylates, simple alkanoates and 1-alkanols, all with chain lengths ranging from C16 to C24. A second fraction, with an average molecular weight about three times higher, was detected by VPO, MS and GPC. The presence of this important fraction in cork suberin had not been recognized in earlier studies. Both fractions constitute interesting precursors for the elaboration of new materials.

Surface Properties of Suberin

The surface energy of suberin was determined by four different methods, namely, (i) contact angle measurements, (ii) Wilhelmy plate measurements, (iii) maximum bubble pressure, and (iv) inverse gas chromatography (IGC). The first three methods gave a gammasub value in the range 40-50 mN m-1 at room temperature. The major component of this value reflects the dispersive contribution. The IGC measurements showed a higher dispersive term, which is common with this method of characterization. The surface acid (A)/base (B) properties were also evaluated, and the results indicated that suberin has an acidic character.

[A rare cause of intestinal obstruction]. / Cause rare di occlusione intestinale.

Three-hundred-thirteen cases of small and large bowel obstructions surgically managed, from May 1974 to September 1995, are reviewed. Particularly, 10 uncommon cases are described: 3 hernias in anomalous recesses, 1 small bowel spontaneous hematoma, 2 Ogilvie syndromes, 1 obstruction due to cholecystitis, 1 left diaphragmatic hernia, 1 encapsulating peritonitis, 1 incarcerated Spigelian hernia. Their aetiopathogenetic and clinical patterns are examined. The Authors conclude, in agreement with Literature, that small bowel obstructions need urgent operation because of high risk of ischemic lesions. In large bowel obstructions operation is suggested after more careful diagnostic researches and adequate patient preparation. In some uncommon obstructions the purpose is to restore normal anatomical conditions.

Effects induced by olive oil-rich diet on erythrocytes membrane lipids and sodium-potassium transports in postmenopausal hypertensive women.

Since we have observed that monounsaturated fatty acids (MUFA) enriched diet modifies red cell membrane lipids and cation transport systems in normotensive subjects, we similarly evaluated a group of hypertensive patients undergoing an analogous dietary modification. In a group of 18 moderately hypertensive women, the diet was supplemented for two months with olive oil (about 45 g/day), which replaced an equal amount of seasoning fats. Before and after this period, red cell fatty acid composition was evaluated by gas-chromatography in order to verify diet compliance: a significant increase in oleic acid was observed, while the content of saturated and polyunsaturated fatty acids remained unchanged. After olive oil, maximal rates of Na-K pump (5580 +/- 329 vs 6995 +/- 390, p less than 0.001) and Na-K cotransport (Na-COT 544 +/- 52 vs 877 +/- 46, p less than 0.001: K-COT 790 +/- 76 vs 1176 +/- 66, p less than 0.001), cell Na content (9.58 +/- 0.4 vs 10.61 +/- 0.6, p less than 0.03) and passive permeability for Na (936 +/- 74 vs 1836 +/- 102, p less than 0.001) rose significantly. Although the reduction in maximal rate of the Li-Na CT after olive oil was not significant, it was the only cation transport parameter being correlated with the variations of membrane lipids, namely negatively with UFA (r = -0.528, p less than 0.05) and positively with SFA (r = 0.482, p less than 0.005). The change in maximal rate of Li-Na CT was also correlated with the variation of systolic and diastolic BP (r = 0.50, p less than 0.03).(ABSTRACT TRUNCATED AT 250 WORDS)

[Chemotaxis deficiency in patients with HIV infection]. / Deficit della chemiotassi in pazienti con infezione da HIV.

Increased susceptibility to bacterial recurrent infection is characteristically associated with impaired B cells function but also with a defective PMN function. We studied PMN CT in 15 HIV positive drug addicts patients with persistent generalized lymphoadenopathy (PGL), in 15 symptom free HIV negative drug addicts (SFDA) and in 15 healthy blood donors to evaluate influence of HIV infection on PMN functions. CT of patients with PGL was reduced to 78% (p < 0.0001) and 75% (p < 0.00001) of CT in SFDA patients and healthy blood donors, respectively. We conclude that HIV infection causes defective PMNL CT and then it can increase susceptibility to bacterial recurrent infections in these patients.

Transmembrane cation fluxes and fatty acid composition of erythrocytes in psoriatic patients.

Cation transport systems and lipid composition of erythrocyte membrane were studied in 27 psoriatic patients and in 34 healthy individuals. Whereas intracellular Na and K content, Na- and K-passive permeability and Li-Na countertransport of psoriatics did not show any statistical difference from normals, the Na/K ATPase pump activity was significantly higher and Na-K cotransport was significantly lower. Membrane lipid composition of psoriatics was different from normals: an increase in arachidonic acid and in unsaturated (poly- and total unsaturated) fatty acid content was found. A positive correlation was demonstrated between unsaturated/saturated fatty acid ratio and Na/K ATPase pump activity. These results demonstrate an alteration of erythrocyte Na/K ATPase pump and Na-K cotransport in psoriasis. These alterations of cation transport are associated with a perturbation of membrane fatty acid composition which appears a widespread phenomenon in cells of psoriatic patients.

Membrane fatty acids and erythrocyte Li-Na countertransport in nephrotic syndrome and their relationship.

Cholesterol/phospholipids molar ratio and fatty acid composition have been estimated in erythrocyte membrane of 12 patients suffering from nephrotic syndrome and compared to values obtained in 23 normal subjects matched for sex and age. The membrane lipid composition has been correlated with the activity of erythrocyte Li-Na countertransport of the same subjects. The results show a significant increase in cholesterol/phospholipids ratio and total saturated fatty acids when erythrocytes of nephrotic patients are compared to normal erythrocytes, whereas total unsaturated fatty acids were lower in nephrotics (p less than 0.002). Li-Na countertransport was higher in nephrotics (p less than 0.001) and it was positively correlated with the total amount of saturated fatty acids of the erythrocyte membrane (r = +0.451; p less than 0.01). On the contrary, Li-Na countertransport was negatively correlated with the total amount of unsaturated fatty acids (r = -0.468; p less than 0.01).