The genetic background and vitamin D supplementation can affect irisin levels in Prader-Willi syndrome.

BACKGROUND: Prader-Willi syndrome (PWS) is associated to distinctive clinical symptoms, including obesity, cognitive and behavioral disorders, and bone impairment. Irisin is a myokine that acts on several target organs including brain adipose tissue and bone. The present study was finalized to explore circulating levels of irisin in children and adult PWS patients. METHODS: Seventy-eight subjects with PWS, 26 children (15 females, mean age 9.48 ± 3.6 years) and 52 adults (30 females, mean age 30.6 ± 10.7) were enrolled. Irisin serum levels were measured in patients and controls. Its levels were related with anthropometric and metabolic parameters, cognitive performance and bone mineral density either in pediatric or adult PWS. Multiple regression analysis was also performed. RESULTS: Irisin serum levels in PWS patients did not show different compared with controls. A more in-depth analysis showed that both pediatric and adult PWS with DEL15 displayed significantly reduced irisin levels compared to controls. Otherwise, no differences in irisin concentration were found in UPD15 patients with respect to controls. Our study revealed that in pediatric PWS the 25(OH) vitamin-D levels affected irisin serum concentration. Indeed, patients who were not supplemented with vitamin D showed lower irisin levels than controls and patients performing the supplementation. Multiple regression analysis showed that irisin levels in pediatric and adult PWS were predicted by the genetic background and 25(OH)-vitamin D levels, whereas in a group of 29 adult PWS also by intelligent quotient. CONCLUSION: We demonstrated the possible role of genetic background and vitamin-D supplementation on irisin serum levels in PWS patients.

CELLULAR MECHANISMS OF BONE REGENERATION: ROLE OF WNT-1 IN BONE-MUSCLE INTERACTION DURING PHYSICAL ACTIVITY39.

Wnt1 is one of the several glycoproteins activating Wnt signaling, critical for normal skeletal development and bone homeostasis. Wnt1 was previously believed to solely regulate central nervous system development, in particular in midbrain and cerebellum. However, remarkable findings have recently shown that several patients affected by severe form of Osteogenesis Imperfecta (OI) display a Wnt1 mutation thereby revealing a possible role of Wnt1 in bone metabolism. Here, we show that recombinant Wnt1 (r-Wnt1) strongly increases differentiation of bone marrow stromal cells into mature osteoblasts, as demonstrated by the enhanced number of cells positively stained for alkaline phosphatase, one of the osteoblastic marker genes, whose mRNA levels are also significantly up-regulated. Furthermore, other osteogenic master genes such as Collagen I and Osteopontin are also enhanced when bone marrow precursors were differentiated toward osteoblastic phenotype in the presence of r-Wnt1. Intriguingly, by in vivo and in vitro findings, we report that in the bone marrow of mice subjected to physical activity there is a high endogenous Wnt1 synthesis compared to mice kept in resting conditions. Moreover, conditioned medium collected from ex vivo myoblasts, harvested from exercised mice, up-regulates Wnt1 expression in osteoblast cell cultures obtained from control mice. Overall our findings support the role of Wnt1 in regulating bone metabolism and suggest that this molecule could be one of the mediators through which physical activity may exert beneficial effect on bone.

Multidetector computed tomography in the study of pancreatic metastases.

PURPOSE: The aim of this study was to explore the diagnostic performance of multidetector computed tomography (MDCT) in characterising pancreatic metastases. MATERIALS AND METHODS: CT examinations of 17 patients affected by pancreatic metastases were retrospectively reviewed. The primary malignancy was renal cell carcinoma (RCC) in eight cases, uterine leiomyosarcoma in two, lung carcinoma in four and breast carcinoma in three. CT images were assessed for lesion number, size and morphology. RESULTS: Pancreatic lesions were solitary in seven cases and multiple in ten. Lesion size ranged between 8 and 40 mm. Metastases from RCC were hyperattenuating in the arterial phase, metastases from breast cancer and lung cancer were hypoattenuating and metastases from uterine leiomyosarcoma were inhomogeneous. Precise lesion characterisation was obtained by using CT examination in 12 cases. In the remaining five patients, all with solitary metastases from RCC, a precise diagnosis was not possible because the lesions could not be differentiated from a neuroendocrine tumour. CONCLUSIONS: MDCT allowed pancreatic metastases characterisation in 70.5% of cases. The lesions were the manifestation of widely disseminated neoplastic disease, with the exception of metastases from RCC, which were exclusively located in the pancreas.

Osteoblast apoptosis in periodontal disease: role of TNF-related apoptosis-inducing ligand.

Periodontal disease (Pd) is characterized by an increased osteoclast resorption and a decreased osteoblast (OB) bone formation. OBs obtained from alveolar bone of Periodontitis patients (Pp) undergo apoptosis in the presence of TNF-related apoptosis-inducing ligand (TRAIL). We studied the intracellular apoptotic pathway induced by TRAIL; TRAIL death (DR4, DR5) and decoy (DcR1, DcR2) receptors expression in Periodontitis patients OBs (PpOBs), and we measured the concentration of TRAIL in the serum of Pp. We demonstrated that DNA fragmentation and activation of caspase-8 and caspase-3 in PpOBs, following TRAIL stimulation, occurred in shorter time; moreover, a higher amount of both caspases was activated in order to direct OBs. Down-regulation of DcR2 in PpOBs was demonstrated and high TRAIL levels were detected in the serum of Pp. In conclusion, our data suggest that PpOBs are more sensitive to TRAIL-induced apoptosis when compared to the control group. The down-regulation of DcR2 possibly leads to an imbalanced ratio between death and decoy receptors. Our findings highlight a role of TRAIL in the pathogenesis of Pd.

Alteration of activity and survival of osteoblasts obtained from human periodontitis patients: role of TRAIL.

Periodontal disease (Pd) is characterized by extensive alveolar bone loss, that occurs as a consequence of the impairment of the normal bone remodelling. Bone remodelling is regulated by the correct balance between osteoclast and osteoblast formation and activity. Alveolar bone loss could be due to an increased bone resorption by osteoclasts or a decreased bone formation by osteoblasts (OBs) or both. Although the role played by osteoclasts in increasing bone resorption in Pd is already known, the behaviour of OBs in this disease is poorly understood. In the present study we hypothesized that activity and survival of OBs, locally present in alveolar bone of Pd patients, are altered. Thus, we studied the activity and survival of OBs obtained from alveolar bone fragments of Pd patients. The results, obtained in OBs from the patients were compared with those from OBs obtained from healthy donors. We demonstrated that OBs from Pd patients weakly express OB phenotype in respect to the control cells. In particular, the alkaline phosphatase activity and the collagen type I production, as well as the formation of mineralized nodules, typical markers of differentiated OBs, were significantly lower in Pd patients. Interestingly, we also demonstrated that OBs from the patients were more sensitive to the apoptotic effect induced by TNF-related apoptosis-inducing ligand (TRAIL). TRAIL, a member of the TNF superfamily, induces apoptosis by interacting with its death receptors, (DR4, DR5). However, its activity can be modulated by two decoy receptors, DcR1 and DcR2. Thus, the sensitiveness of TRAIL induced apoptosis is determined by the ratio of death and decoy receptor. We demonstrated that OBs from Pd patients showed an imbalanced ratio between death and decoy TRAIL receptors due to the down-regulation of DcR2 expression. Furthermore, the levels of TRAIL in the serum of the same patients were significantly higher than those detected in the controls. In conclusion, we show for the first time that the alveolar bone loss in Pd patients could be due to the increased TRAIL-mediated apoptosis of OBs.

T cells support osteoclastogenesis in an in vitro model derived from human periodontitis patients.

BACKGROUND: Periodontitis is characterized by alveolar bone destruction; however, the mechanisms responsible for bone damage are poorly understood. It has been reported that T cells are implicated in the pathogenesis of periodontitis. It has been also demonstrated that activated T lymphocytes secrete receptor activator of nuclear factor-kappa B ligand (RANKL) and can support the differentiation of monocytes into resorbing osteoclasts (OCs). Therefore, the purpose of this study was to examine the OC formation in periodontitis patients (PP) and the role of T cells in osteoclastogenesis. METHODS: To study OC formation, we used an in vitro model consisting of unstimulated and unfractionated peripheral blood mononuclear cells (PBMCs) from PP and controls. In parallel, T-cell-depleted PBMCs from the same patients were also established. The expression of RANKL and tumor necrosis factor-alpha (TNF-alpha) was analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot in fresh T cells isolated from PP and controls. Functional antibodies, anti-RANKL and anti-TNF-alpha, were utilized to study osteoclastogenesis in PBMC cultures from PP. RESULTS: We showed that, in unfractionated PBMCs from PP, the OCs spontaneously developed in a T-cell-dependent way. The addition of macrophage colony stimulating factor (MCSF) and RANKL was necessary to promote the osteoclastogenesis in T-cell-depleted PBMC cultures from PP and in unfractionated PBMCs from periodontally healthy controls. Moreover, freshly isolated T cells from PBMCs of PP overexpressed RANKL and TNF-alpha. Finally, functional anti-RANKL and anti-TNF-alpha antibodies significantly inhibited osteoclastogenesis. CONCLUSION: Our data suggest that T cells support spontaneous osteoclastogenesis in PP via RANKL and TNF-alpha overexpression.

Interleukin-7 production by B lymphocytes affects the T cell-dependent osteoclast formation in an in vitro model derived from human periodontitis patients.

Periodontitis is characterized by irreversible destruction of alveolar bone and connective tissue attachment in the periodontium. We recently reported that T cells support the osteoclastogenesis by the overproduction of nuclear factor-kappa-B-ligand (RANKL) and Tumor Necrosis Factor-alpha (TNF-alpha) in an in vitro osteoclastogenesis model from periodontitis patients (Pp). It is known that IL-7 stimulates the production of osteoclastogenic factors by T cells and IL-6 potentiates IL-7 expression. Thus, we studied the involvement of IL-6 and IL-7 in the T cell regulation of osteoclast (OC) formation, in an in vitro osteoclastogenesis model from Pp. We demonstrated high levels of IL-7 in both the media collected from PBMC cultures of Pp and the sera of the same patients. We also demonstrated that freshly isolated B cells from PBMCs of Pp were the source of IL-7 in our model. B cells, in fact, overexpressed IL-7 at mRNA and protein levels, and this production was up-regulated by IL-6. Moreover, the OC formation decreased in the presence of anti-IL-6 and IL-7 functional antibodies in PBMC cultures from Pp. These data suggest that B cells could be responsible for the T cell-dependent osteoclastogenesis in periodontitis through the involvement of IL-6 and IL-7.

Leprosy and AIDS: two cases of increasing inflammatory reactions at the start of highly active antiretroviral therapy.

Reported here are the cases of two HIV-positive patients with skin lesions suggestive of leprosy, based on clinical and pathological analysis, which worsened during the few weeks following initiation of highly active antiretroviral therapy. The lesions improved after a few weeks of multidrug therapy for leprosy. Mycobacterium leprae was confirmed by polymerase chain reaction analysis of blood in case 1 and of a biopsy sample in case 2. Neither Mycobacterium avium complex nucleic acid, which is usually associated with immune restoration syndrome, nor mycobacterial cutaneous manifestations were detected in either case.

Leprosy and AIDS: two cases of increasing inflammatory reactions at the start of highly active antiretroviral therapy

Reported here are the cases of two HIV-positive patients with skin lesions suggestive of leprosy, based on clinical and pathological analysis, which worsened during the few weeks following initiation of highly active antiretroviral therapy. The lesions improved after a few weeks of multidrug therapy for leprosy. Mycobacterium leprae was confirmed by polymerase chain reaction analysis of blood in case 1 and of a biopsy sample in case 2. Neither Mycobacterium avium complex nucleic acid, which is usually associated with immune restoration syndrome, nor mycobacterial cutaneous manifestations were detected in either case

Distinct histopathological patterns in single lesion leprosy patients treated with single dose therapy (ROM) in the Brazilian Multicentric Study.

This paper aims to describe the histomorphologic features of skin biopsies of single lesion leprosy patients recruited at outpatient clinics in four Brazilian states in the Northeast (Amazonas and Rondonia), Southeast (Rio de Janeiro) and Center-West (Goiás) between October 1997 and December 1998. Patients clinically diagnosed as single skin lesion paucibacillary (SSL-PB) leprosy had a standard 4-mm punch biopsy taken from the lesion before rifampin, ofloxacin, minocycline (ROM) therapy. The features of the cellular inflammatory infiltrates, the presence of nerve involvement and acid-fast bacilli (AFB) were used to categorize SSL-PB biopsies into different histopathological groups. Two-hundred-seventy-eight (93.0%) out of 299 patients had a skin biopsy available. Seven single lesion patients were diagnosed as BL or LL leprosy types (MB) by the histopathological exams and 12 cases were excluded due to other skin diseases. Therefore, 259 patients had skin lesions with histomorphological features compatible with PB leprosy categorized as follows: 33.6% (N = 87) of the biopsies represented well-circumscribed epithelioid cell granuloma (Group 1); 21.6% (N = 56) less-circumscribed epithelioid cell granuloma (Group 2); 12.0% (N = 31) were described as mononuclear inflammatory infiltrate permeated with epithelioid cells (Group 3), and 29.7% (N = 77) had perivascular/periadnexal mononuclear inflammatory infiltrate (Group 4). Minimal/no morphological alteration in the skin was detected in only 8 (3.1%) SSL-PB patients categorized as Group 5, who were considered to have leprosy by clinical parameters. SSL-PB leprosy patients recruited in a multicentric study presented histomorphology readings comprising the whole PB leprosy spectrum but also a few MB cases. These results indicate heterogeneity among SSL-PB patients, with a predominance of well-circumscribed and less-circumscribed epithelioid cell granulomas (Groups 1 and 2) in the sites studied and the heterogeneity of local cellular immune response.

Tuberculose ganglionar em pacientes co-infectados pelo HIV-1: estudo clínico e laboratorial / Ganglionic tuberculosis in patients co-infected by HIV-1: clinical and laboratorial study

Estudos clínicos e laboratoriais apontam diferenças marcantes entre pacientes soropositivos para HIV-1 e imunocompetentes. A tuberculose ganglionar é uma forma freqüente de apresentaçäo extrapulmonar nestes pacientes. A apresentaçäo histopatológica é muito diferente dos pacientes imunocompetentes, com mais de 70 por cento dos casos de várias séries näo apresentando granulomas ou, quando presentes, malformados, com extensa necrose caseosa, raras células gigantes e presença de muitos bacilos álcool-ácido resistentes, denotando uma resposta imune incompleta. Estudamos os diversos aspectos clínicos e laboratoriais, incluídos exames microbiológicos e histopatológicos de 27 pacientes encaminhados para tratamento e diagnóstico no Hospital Evandro Chagas/FIOCRUZ. De acordo com o diagnóstico histopatológico, os casos foram subdivididos em três grupos: Grupo I (n = 7): tuberculose ganglionar sem infecçäo pelo HIV - 1; Grupo II (n = 9): tuberculose ganglionar em pacientes co-infectados por HIV - 1; Grupo III (n = 11): linfadenite reativa associada à AIDS. No grupo de pacientes co-infectados em relaçäo aos imunocompetentes, verificou-se grannulomas malformados, verificou-se granulomas malformados, extensa necrose e grande riqueza bacilar, além de queda acentuada de hemácias e linfócitos e grande freqüência de infecçöes oportunísticas denotando grave defeito na resposta imune, explicando a apresentaçäo de formas bem mais severas e nao usuais de tuberculose nos pacientes com AIDS.

[Crohn's disease of the appendix: a rare nosological entity]. / Il morbo di Crohn dell'appendice: una rara entità nosologica.

In this study, the authors describe a case of Crohn's disease confined to the vermiform appendix out of a total of 1,263 appendectomies performed over the past 10 years in a Clinical Surgery Department (Clinica Chirurgica III) of the Policlinico S. Orsola in Bologna. In agreement with other reports in the scientific literature, the essentially non-specific clinical picture prevents pre-operative identification of the condition, whereas a diagnosis of Crohn's disease can be reached only on the basis of histological characteristics. The extreme rarity of Crohn's disease confined to the appendix is, however, confirmed. Follow-up at 3 years revealed no involvement of other areas of the digestive tract.

[Sulpiride and extrahepatic bile ducts. Radioisotope study]. / Sulpiride e vie biliari extraepatiche. Ricerca con radioisotopi.

An original radioisotopic method was employed in an investigation of the effect of sulpiride on extrahepatic bile duct motility. Stress is laid on the primarily hyposthenic effect of the drug, and the influence of various factors on the functional status of the gallbladder.

[Malate dehydrogenase and glutamate dehydrogenase activity modification in livers of rats exposed to a double stress (dry heat - fasting). I]. / Modificazioni dell'attivita' della malico-deidrogenase e della glutammato-deidrogenasi nel fegato di ratti sottoposti a duplice stress (caldo secco - digiuno). Nota I.

We have studied MDH and GLDH behaviour in liver homogenized of rats exposed to double stress (dry heat and starvation) for 24 hours respectively. Both enzyme showed a statistically significant activity increase, either after 24 and 72 hours. Such an increase is affected mainly by starvation. This behaviour reflects the metabolic needs of organism, related to increased energetic requirements.

[Modification of malate dehydrogenase and glutamate dehydrogenase in kidneys of rats exposed to a double stress (dry heat - fasting). II]. / Modificazioni dell'attivita' della malico-deidrogenasi e della glutammato-deidrogenasi nel rene di ratti sottoposti a duplice stress (caldo secco - digiuno). Nota II.

MDH and GLDH activity changes have been studied in kidney homogenized of rats exposed to double stress (dry heat and starvation) for 24 and 72 hours respectively. MDH behaviour activity decrease after 24 hours and increase to basic values after 72 hours leads to suspect the primer of a defence mechanism that precedes to face up increased metabolic needs. GLDH activity shows a constant decrease after 24 hours and after 72 hours; this was explained as a premature acclimatization reaction.

[Modification of malate dehydrogenase and glutamate dehydrogenase in hearts of rats exposed to a double stress (dry heat - fasting). III]. / Modificazioni dell'attivita' della malico-deidrogenasi i della glutammato-deidrogenasi nel cuore di ratti sottoposti a duplice stress (caldo secco - digiuno). Nota III.

MDH and GLDH activity modifications have been studied in heart homogenized of rats exposed to double stress (dry heat and starvation). MDH activity shows statistically significant variations only after 72 hours. This induces to suspect a decrease of energetic metabolism due to a probable substratum lack. GLDH activity shows a statistically significant decrease after 24 hours, and this decrease increases after 72 hours. This behaviour could be explained by an acclimatization reaction of such an amount to delete the hunger effect on protein catabolism.