Preparation of enriched populations of corticotrophs from goldfish rostral pars distalis

A mammalian isolated adrenal cell system was validated as a bioassay for goldfish ACTH; the log dose-response curve for the goldfish hormone is parallel to that for synthetic mammalian ACTH1-24, and the two ACTHs induce the same maximum rate of corticosterone production. Using this assay, it was observed that (1) there is a marked and consistent biphasic change in pituitary ACTH content as related to the length of time the fish are held in laboratory aquaria, and (2) the absolute and relative (to tissue wt) ACTH content of the rostral pars distalis is considerably greater than that of the proximal portion of the gland. Using a trypsin technique, isolated pituitary cells were prepared from both (rostral and proximal) portions of the gland; bioassay data indicate that cell suspensions prepared from the rostral pars distalis are enriched with respect to corticotrophs. The implications of these findings with regard to formulating an advantageous in vitro system for studying ACTH secretion are discussed.     

The impact of next-generation sequencing on the diagnosis of pediatric-onset hereditary spastic paraplegias: new genotype-phenotype correlations for rare HSP-related genes

Hereditary spastic paraplegias (HSP) are clinical and genetic heterogeneous diseases with more than 80 disease genes identified thus far. Studies on large cohorts of HSP patients showed that, by means of current technologies, the percentage of genetically solved cases is close to 50%. Notably, the percentage of molecularly confirmed diagnoses decreases significantly in sporadic patients. To describe our diagnostic molecular genetic approach on patients with pediatric-onset pure and complex HSP, 47 subjects with HSP underwent molecular screening of 113 known and candidate disease genes by targeted capture and massively parallel sequencing. Negative cases were successively analyzed by multiplex ligation-dependent probe amplification (MLPA) analysis for the SPAST gene and high-resolution SNP array analysis for genome-wide CNV detection. Diagnosis was molecularly confirmed in 29 out of 47 (62%) patients, most of whom had clinical diagnosis of cHSP. Although SPG11 and SPG4 remain the most frequent cause of, respectively, complex and pure HSP, a large number of pathogenic variants were disclosed in POLR3A, FA2H, DDHD2, ATP2B4, ENTPD1, ERLIN2, CAPN1, ALS2, ADAR1, RNASEH2B, TUBB4A, ATL1, and KIF1A. In a subset of these disease genes, phenotypic expansion and novel genotype-phenotype correlations were recognized. Notably, SNP array analysis did not provide any significant contribution in increasing the diagnostic yield. Our findings document the high diagnostic yield of targeted sequencing for patients with pediatric-onset, complex, and pure HSP. MLPA for SPAST and SNP array should be limited to properly selected cases based on clinical suspicion.     

Absence of filaggrin mutation in a patient affected by pachyonychia congenita and mild atopic dermatitis

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Traumatic brain injury and hypopituitarism in children and adolescents: is the problem under-estimated?

Traumatic brain injury (TBI) is the leading cause of death in children. Only recently, the importance of hypopituitarism as a consequence of TBI has been highlighted in adult patients. Data from systematic clinical studies in adults and patients in transition from adolescence to adulthood point to the presence of hypopituitarism in nearly a third of patients hospitalized for TBI. But, no systematic studies on posttraumatic hypopituitarism exist in children and adolescents. Case reports and small series of patients, however, suggest that children are affected to a comparable extent. Since normal pituitary function is required for normal growth and pubertal development in childhood, particular attention should be paid to this condition. The aim of this review is to summarize the literature on TBI and causes, clinical picture, and diagnosis of hypopituitarism in childhood and adolescence, underlying the relevance of the problem and its underestimation in clinical paediatric practice.     

Technical and clinical feasibility of contrast-enhanced ultrasound evaluation of long bone non-infected nonunion healing

Purpose

To assess the technical feasibility of contrast-enhanced ultrasound (CEUS) in the monitoring of non-infected long bone nonunion healing.

Methods

Twenty-five patients (16 males; mean age: 40.4?±?11.7) with long bone nonunion were treated using surgery and mesenchymal stem cells and platelet-rich plasma. They performed CEUS up to 15 days before, 7 days, 4 and 8 weeks after treatment. To categorize the angiogenesis around the fracture site, the microvascular blood flow from CEUS was classified into four categories, depending on the portion of the investigated area that was involved in the neovascularization process: grade 0?=?0%; grade 1?=?0–30%; grade 2?=?30–70%; grade 3?=?70–100%. Nonparametric Friedman and Wilcoxon statistics were used.

Results

Before treatment, neovascularization was graded as 0 in 15/25 patients, as 1 in 10/25. Vascularity significantly increased over time (P?<?0.001), namely: 1 (25th–75th percentile?=?1–2) at 7 days; 2 (1–2) at 4 weeks; 3 (0–2) at 8 weeks. All patients but one showed early progressive increase in neovascularization well identified with CEUS at the fracture site.

Conclusion

CEUS is a feasible method to monitor healing in patients with long bone nonunion.

     

Targeting endothelial-to-mesenchymal transition: the protective role of hydroxytyrosol sulfate metabolite

European Journal of Nutrition - Endothelial-to-mesenchymal transition (EndMT) plays an important role in pathogenesis of a number of inflammatory diseases. Hydroxytyrosol (HT) and, particularly,...