"Implicazioni epidemiologiche e terapeutiche della variabilita genetica di HIV-1"

Gli obiettivi principali del lavoro svolto in questi ultimi tre anni sono stati il monitoraggio dell’andamento delle farmacoresistenze, con la definizione di quali fattori siano associati al rischio di sviluppo di resistenza nella pratica clinica; la circolazione dei sottotipi non-B e la caratterizzazione di alcuni isolati di particolare interesse; la distribuzione delle mutazioni di resistenza nel DNA linfomonocitario vs. l’RNA plasmatico in soggetti ‘naive’; il monitoraggio delle variazioni aminoacidiche nel gene env al fine di valutare la suscettibilità ad uno degli ultimi farmaci introdotti nella terapia antiretrovirale, l’inibitore di fusione T20. E’ ,inoltre, in corso l’allestimento di un saggio per lo studio della fitness replicativa basato sulla ricombinazione fra un frammento genomico ottenuto dalla popolazione virale del paziente e il ‘backbone’ di HIV-1 pNL4-3. Il saggio di fitness consisterà in una coltivazione competitiva dell’ibrido così costruito e del tipo selvaggio di riferimento reso riconoscibile mediante inserimento di mutazioni silenti nella regione gag. La dinamica del rapporto quantitativo fra i due virus verrà analizzata a una serie di punti temporali tramite impiego di una coppia di ‘probes’ in grado di distinguere il virus selvaggio dal ricombinante in esame

Sarcoidosis presenting with splenomegaly and abdominal pain: a case report

Sarcoidosis is a multisystemic disease of unknown aetiology characterized by proliferation of noncaseating granulomas at disease sites. It commonly affects young and middle-age adults and frequently presents with pulmonary infiltration, bilateral hilar lymphadenopathy, ocular and skin lesions. The liver, spleen, lymph nodes, salivary glands, heart, nervous system, muscles, bones, and other organs may also be involved. A diagnosis of the disorder usually requires the demonstration of typical lesions in more than one organ system and exclusion of other disorder known to cause granulomatous inflammation. We present the case of a young woman with abdominal pain and weight loss. The finding of splenomegaly by abdominal ultrasound and the presence of hypercalciuria, hypercalcemia and mild renal impairment led us to include sarcoidosis in the differential diagnosis. The final diagnosis was established by demonstration of involvement of lymph nodes and lung parenchyma on CT scan, and typical histology in bioptic specimens collected from bronchial mucosa

Main-duct intraductal papillary mucinous neoplasm of the pancreas: a case report

Three distinct entities among non-inflammatory cystic lesions of the pancreas have been defined: intraductal papillary mucinous neoplasm (IPMN), serous cystic neoplasm (SCN) and mucinous cystic neoplasm (MCN). IPMN is characterized by intraductal papillary growth and thick mucus secretion: its incidence has dramatically increased since its initial description. These lesions probably can progress towards invasive carcinoma. IPMNs are symptomatic in most cases: the typical presentation is a recurrent acute pancreatitis, without evident cause, of low or moderate severity. The diagnosis is usually based upon the imaging (CT/cholangio-MRI) demonstrating a pancreatic cystic mass, involving a dilated main duct, eventually associated to some filling defects, or a normal Wirsung duct communicating with the cyst lesion. Surgical treatment is generally indicated for main duct IPMN and branch duct IPMN with suspected malignancy (tumour size ≥ 30 mm, mural nodules, dilated main pancreatic duct, or positive cytology) or prominent symptoms. Herein we present a case of IPMN of the main duct which occurred with abdominal and back pain associated with weight loss. After the diagnosis, she successfully underwent surgery and is now in a follow-up program

Comparative determination of HIV-1 co-receptor tropism by Enhanced Sensitivity Trofile, gp120 V3-loop RNA and DNA genotyping

BACKGROUND: Trofile is the prospectively validated HIV-1 tropism assay. Its use is limited by high costs, long turn-around time, and inability to test patients with very low or undetectable viremia. We aimed at assessing the efficiency of population genotypic assays based on gp120 V3-loop sequencing for the determination of tropism in plasma viral RNA and in whole-blood viral DNA. Contemporary and follow-up plasma and whole-blood samples from patients undergoing tropism testing via the enhanced sensitivity Trofile (ESTA) were collected. Clinical and clonal geno2pheno[coreceptor] (G2P) models at 10% and at optimised 5.7% false positive rate cutoff were evaluated using viral DNA and RNA samples, compared against each other and ESTA, using Cohen's kappa, phylogenetic analysis, and area under the receiver operating characteristic (AUROC). RESULTS: Both clinical and clonal G2P (with different false positive rates) showed good performances in predicting the ESTA outcome (for V3 RNA-based clinical G2P at 10% false positive rate AUROC = 0.83, sensitivity = 90%, specificity = 75%). The rate of agreement between DNA- and RNA-based clinical G2P was fair (kappa = 0.74, p < 0.0001), and DNA-based clinical G2P accurately predicted the plasma ESTA (AUROC = 0.86). Significant differences in the viral populations were detected when comparing inter/intra patient diversity of viral DNA with RNA sequences. CONCLUSIONS: Plasma HIV RNA or whole-blood HIV DNA V3-loop sequencing interpreted with clinical G2P is cheap and can be a good surrogate for ESTA. Although there may be differences among viral RNA and DNA populations in the same host, DNA-based G2P may be used as an indication of viral tropism in patients with undetectable plasma viremia

Use of peripheral blood DNA for genotypic antiretroviral resistance testing in drug-naïve HIV-infected subjects.

Parallel analysis of peripheral blood mononuclear cell DNA and plasma RNA from 169 drug-naive human immunodeficiency virus-infected subjects revealed that evaluation of both compartments increases the sensitivity of detection of drug resistance-related mutations, compared with examination of either source alone. Peripheral blood mononuclear cell DNA may play a role in the surveillance of transmitted antiretroviral resistance

NEUROHORMONES IN MITRAL STENOSIS BEFORE AND AFTER PERCUTANEOUS BALLOON MITRAL VALVOTOMY

BACKGROUND AND AIM OF THE STUDY: The hormonal response to percutaneous balloon mitral valvotomy (PBMV) has been described in patients in sinus rhythm (SR) and with atrial fibrillation (AF). The study aim was to evaluate the effect of hemodynamic parameters and PBMV on atrial natriuretic factor (ANF) secretion and plasma renin activity (PRA) in mitral stenosis in SR and AF. METHODS: Thirty-one patients (26 females, five males; mean age 50.5+/-14 years) with pure rheumatic mitral stenosis underwent PBMV. Fourteen patients had AF, and 17 were in SR. PRA and ANF were measured 24 h before, and at 30 and 60 min, 24 h and one month after PBMV, after resting in a supine position for > or =2 h. Digitalis and diuretics were withdrawn 48 h before sampling; neither had patients received ACE inhibitors or beta-blockers during the previous month. RESULTS: PBMV was successful in all cases, without complication. Mitral valve area was increased and wedge pressure decreased in both groups after PBMV. In AF patients, neither PRA nor ANF were significantly affected before and after PBMV; in SR patients, ANF was decreased and PRA increased significantly, notably 24 h after PBMV. The cardiac index was increased in both groups, but was distinctly lower in AF patients both before and after PBMV. CONCLUSION: Despite similar hemodynamic results, reversal of the hormonal pattern after PBMV occurred only in SR patients, most likely because in AF patients a low cardiac index elicits a hormonal response similar to heart failure. This abnormal hormonal pattern may limit functional recovery after PBMV; hence, PBMV is best attempted while patients are still in SR

Three-Class-Resistant Human Immunodeficiency Virus Type 1 Variant in a Drug-Naive Heterosexual Couple▿

Combination antiretroviral treatment was initiated in a heterosexual couple newly diagnosed with human immunodeficiency virus type 1 infection. Multiple genotypic drug resistance testing following early rebound of viral load revealed that the same three-class-resistant human immunodeficiency virus type 1 strain had been present in both patients since before initiation of treatment