Seasonal changes in the liver of a non-hibernating population of water frogs, Pelophylax kl. esculentus (Anura: Ranidae)

Seasonal variation of liver glycogen, lipids and melanomacrophages were investigated in a non-hibernating population of Pelophylax kl. esculentus from Calabria by histochemical methods and computer-assisted image analysis. Twenty individuals of both sexes were sampled in a tank in Roseto Capo Spulico (Cosenza, Calabria) in four periods of the year 2016 (February, May, July, October). Portions of liver from each individual were included in paraffin for glycogen and melanomacrophages, and epoxydic resin-araldite for lipid analysis. Sections were stained with periodic acid-Schiff (PAS) for glycogen (with diastase-PAS as control) or osmium-tetroxide for lipids, or left unstained for melanomacrophages (appearing naturally black due to melanin). Image analyses were performed on 9–12 grayscale converted pictures per individual. Total areas per µm2 of glycogen, lipids and melanomacrophages, as well as counts of lipid droplets and melanomacrophages and mean area of single lipid droplets and melanomacrophages, were measured. Statistical analyses were performed by analysis of variance (ANOVA) with bootstrap resampling. Significant variation among sampling periods was found for each variable. Glycogen and lipids co-vary, with higher values observed in October–February and lower values in May–July, whereas melanomacrophages reach a peak in May and have much lower values in the other months. It is concluded that, in the absence of a hibernating period, reproduction is the main force regulating the annual cycles of reserve storing and melanin production

Gene electrotransfer of IL-2 and IL-12 plasmids effectively eradicated murine B16.F10 melanoma

Gene therapy has become an important approach for treating cancer, and electroporation represents a technology for introducing therapeutic genes into a cell. An example of cancer gene therapy relying on gene electrotransfer is the use of immunomodulatory cytokines, such as interleukin 2 (IL-2) and 12 (IL-12), which directly stimulate immune cells at the tumour site. The aim of our study was to determine the effects of gene electrotransfer with two plasmids encoding IL-2 and IL-12 in vitro and in vivo. Two different pulse protocols, known as EP1 (600 V/cm, 5 ms, 1 Hz, 8 pulses) and EP2 (1300 V/cm, 100 µs, 1 Hz, 8 pulses), were assessed in vitro for application in subsequent in vivo experiments. In the in vivo experiment, gene electrotransfer of pIL-2 and pIL-12 using the EP1 protocol was performed in B16.F10 murine melanoma. Combined treatment of tumours using pIL2 and pIL12 induced significant tumour growth delay and 71% complete tumour regression. Furthermore, in tumours coexpressing IL-2 and IL-12, increased accumulation of dendritic cells and M1 macrophages was obtained along with the activation of proinflammatory signals, resulting in CD4 + and CD8 + T-lymphocyte recruitment and immune memory development in the mice. In conclusion, we demonstrated high antitumour efficacy of combined IL-2 and IL-12 gene electrotransfer protocols in low-immunogenicity murine B16.F10 melanoma

Morpho-anatomical and microbiological analysis of kiwifruit roots with KVDS symptoms

Italy, one of the largest producers of kiwifruit in the world, has lost 10% of its production in recent years because of the spread of the kiwifruit vine decline syndrome (KVDS). Although the aetiology of KVDS has not been characterized, root rot symptoms are often associated with water stagnation and root asphyxia. To investigate causal factors and potential solutions to counter this syndrome, an experimental trial was undertaken in a kiwifruit orchard affected by KVDS in Latina (central Italy) in 2020. Root samples from healthy plants were collected and compared with samples taken from plants affected by KVDS. Macroscopically, the roots affected by KVDS were rotting, showing a loss of rhizodermis and cortical parenchyma. Microscopic analysis revealed damage to the root system with tissue breakdown and decomposition, flaking of the rhizodermis, cortical area with a clear loss of cell turgor, initial decay of the stele and evident detachment of the cortex from the central conducting tissues. Light microscopy, morphological and molecular analyses were carried out on the rhizodermis of roots showing decay and death symptoms. Total DNA extracted from the pure fungal colonies was amplified by PCR with ITS primers, amplicons directly sequenced, and the obtained nucleotide sequences were compared with those present in the GenBank database (NCBI) through BLAST analysis. Genomic analysis allowed the identification of three abundant fungi namely Ilyonectria vredenhoekensis, Fusarium oxysporum and Paraphaeosphaeria michotii. Further investigation is required to determine the role of these fungi in KVDS, whether they are species favoured by water stagnation and root asphyxia; their abundance and presence in other regions, orchards, and kiwifruit species; if they compromise roots functionality individually or conjunction with other microbial pathogens or abiotic factors; and if they contribute to plant death associated with KVDS

Spinal Volumetric Bone Mineral Density and Vertebral Fractures in Female Patients with Adrenal Incidentalomas: The Effects of Subclinical Hypercortisolism and Gonadal Status

Although adrenal incidentalomas (AI) are not associated with clinically evident syndromes, some patients display biochemical features of subclinical hypercortisolism (SH). Previous studies indicated a negative effect of SH on bone in AI patients, but the prevalence of vertebral fractures and the roles of SH and gonadal status in volumetric bone mineral density are unknown. In 70 female AI patients and 84 controls, the prevalence of vertebral fractures and spinal bone mineral density (by quantitative computed tomography) were evaluated. Subjects were subdivided according to menopausal status into groups Pre (21 patients and 23 controls) and Post (49 patients and 61 controls); there were 14 and 35 patients without SH (SH-) and 7 and 14 patients with SH (SH+) in groups Pre and Post, respectively. The prevalence of fractures was higher in SH+ than in controls and in SH- subjects in both groups Pre [SH+, 42.9%; controls, 0% (P = 0.001); SH-, 7.1% (P = 0.049)] and post [SH+, 78.6%; controls, 37.7% (P = 0.006); SH- 42.9% (P = 0.024)]. In group Post, the mean z-score quantitative computed tomography values were lower in SH+ patients (-0.78 \ub1 0.29) than in controls (0.06 \ub1 0.14; P = 0.011) and SH- patients (0.02 \ub1 0.19; P = 0.034). Evaluation of spinal bone is indicated in female AI patients with SH

Validation of the clinical consensus recommendations on the management of fracture risk in postmenopausal women with type 2 diabetes

Background and aims: Bone fragility is recognized as a complication of type 2 diabetes (T2D). However, the fracture risk in T2D is underestimated using the classical assessment tools. An expert panel suggested the diagnostic approaches for the detection of T2D patients worthy of bone-active treatment. The aim of the study was to apply these algorithms to a cohort of T2D women to validate them in clinical practice. Methods and results: The presence of T2D-specific fracture risk factors (T2D ≥ 10 years, ≥1 T2D complications, insulin or thiazolidinedione use, poor glycaemic control) was assessed at baseline in 107 postmenopausal T2D women. In all patients at baseline and in 34 patients after a median follow-up of 60.2 months we retrospectively evaluated bone mineral density and clinical and morphometric vertebral fractures. No patient was treated with bone-active drug. Following the protocols, 34 (31.8%) and 73 (68.2%) patients would have been pharmacologically and conservatively treated, respectively. Among 49 patients without both clinical fractures and major T2D-related risk factors, who would have been, therefore, conservatively followed-up without vertebral fracture assessment, only one showed a prevalent vertebral fracture (sensitivity 90%, negative predictive value 98%). The two patients who experienced an incident fracture would have been pharmacologically treated at baseline. Conclusions: The clinical consensus recommendations showed a very good sensitivity in identifying T2D postmenopausal women at high fracture risk. Among those with treatment indication as many as 13% of patients experienced an incident fracture, and, conversely, among those without treatment indication no incident fractures were observed

Altered bone mass and turnover in female patients with adrenal incidentaloma: The effect of subclinical hypercortisolism

The strategy of treatment for patients with adrenal incidentalomas (AI) may depend upon the presence of hormonal hypersecretion. Although alterations of bone turnover have been recently reported, data on bone mineral density (BMD) are not available in AI patients. We evaluated bone turnover and BMD in 32 female AI patients and 64 matched controls. Spinal and femoral BMD were similar in patients and controls. Serum bone GLA protein (6.8 \ub1 3.5 vs. 8.8 \ub1 3.2 ng/mL; P < 0.005) and PTH (48.8 \ub1 15.1 vs. 37.2 \ub1 10.9 pg/mL; P < 0.0001) were different in patients and controls. Patients were then subdivided into 2 groups: with (n = 8; group A) or without (n = 24; group B) subclinical hypercortisolism. PTH was higher (P < 0.05) in group A than in group B and in both groups than in controls (57.1 \ub1 13.6, 46.0 \ub1 14.8, and 37.2 \ub1 10.9 pg/mL, respectively), and bone GLA protein was lower in group A than in group B and controls (3.8 \ub1 2.3, 7.5 \ub1 3.1, and 8.8 \ub1 3.2 ng/mL, respectively; P < 0.05). Serum type I cross-linked C telopeptide and fasting urinary deoxypyridinoline/creatinine were not different in the three groups. BMD at each site was lower (P < 0.05) in group A than in group B and controls. Bone mass and metabolism are altered in AI patients with subclinical hypercortisolism and should be taken into account, therefore, when addressing the treatment of choice for these patients

Correction to: Multiple endocrine neoplasia type 1: analysis of germline MEN1 mutations in the Italian multicenter MEN1 patient database.

The original version of this article unfortunately contained a mistake in Table 2. The table 2 was truncated in the original publication. The full table 2 is given below

Threatened and extinct amphibians and reptiles in Italian natural history collections are useful conservation tools

Natural history museums are irreplaceable tools to study and preserve the biological diversity around the globe and among the primary actors in the recognition of species and the logical repositories for their type specimens. In this paper we surveyed the consistency of the preserved specimens of amphibians and reptiles housed in the major Italian scientific collections, and verified the presence of threatened species according to the IUCN Red List, includ-ing the Extinct (EX), Extinct in the Wild (EW), Critically Endangered (CR), Endangered (EN), and Vulnerable (VU) categories. Altogether, we analyzed 39 Italian zoological collections. We confirmed the presence of one extinct reptile (Chioninia coctei) and five extinct or extinct in the wild amphibian species (Atelopus longirostris, Nectophrynoides asperginis, Pseudophilautus leucorhinus, P. nasutus, and P. variabilis). Seven CR amphibians, fourteen CR reptile species and the extinct skink C. coctei are shared by more than one institution. Museums which host the highest number of threatened and extinct amphibian species are respectively Turin (17 CR and 1 EX), Florence (13 CR and 1 EX), and Trento (15 CR and 1 EW), while for reptiles the richest museums are those from Genoa (15 CR and 1 EX), Florence (11 CR and 1 EX), and Pisa (7 CR). Finally, we discussed the utility of natural history museums and the strategies to follow for the implementation of their functionality. © Firenze University Press

Bone involvement in eugonadal male patients with adrenal incidentaloma and subclinical hypercortisolism

Adrenal incidentalomas (AI) are not associated, by definition, with clinically evident syndromes; however, some AI patients may show biochemical indexes of subclinical hypercortisolism (SH). Previous data on female AI patients indicated that SH may lead to bone loss, at least at spine. No data are available on bone involvement in samples of only AI male patients. We measured bone metabolism and bone mineral density at spine and femur by dual-energy x-ray absorptiometry in 38 consecutive eugonadal male AI patients and 38 healthy matched control subjects. Patients were subdivided according to the presence or absence of SH (group SH+ and group SH-, respectively). Mean Z-score levels of spinal bone mineral density measured by dual-energy x-ray absorptiometry were lower (P < 0.05) in group SH+ (-0.42 \ub1 1.62) in comparison with group SH- (0.6 \ub1 1.13) and controls (0.47 \ub1 1.06). Thus, in order for the most appropriate management to be individually tailored, bone mass evaluation is strongly indicated in AI male patients with SH, irrespective of their gonadal status