V348I mutation in UL23 gene of human herpesvirus 1 in a case of herpetic hepatitis and haemophagocytic lymphohistiocytosis.

We herein report the case of a young immunocompetent adult patient with a rapidly fatal haemophagocytic lymphohistiocytosis syndrome related to human herpesvirus 1 (HHV-1) infection, with herpetic hepatitis and persistent high-level viraemia despite treatment with acyclovir. Haemophagocytic lymphohistiocytosis was confirmed in the patient's spleen and bone marrow. HHV-1 DNA was extracted from whole blood and liver biopsy and the UL23 gene was sequenced. A V348I natural polymorphism of the TK protein was found in blood and liver specimens. Further studies are needed to investigate the role of this polymorphism in the development of systemic immune dysregulation.

Detection of antibiotic resistance genes from blood cultures: performance assessment and potential impact on antibiotic therapy management.

Molecular assays may constitute a valid method for timely prediction of antimicrobial resistance and optimization of empirical antibiotic therapies. This study assessed ELITe MGB assays of blood cultures to detect the main carbapenemase and extended-spectrum beta-lactamase (ESBL) genes, Staphylococcus aureus and mec genes in less than 3 h. Excellent agreement was found between the results of genotypic and conventional phenotypic approaches. Retrospective analysis of medical records revealed that approximately 50% of bloodstream infections caused by ESBL-producing Enterobacteriaceae, carbapenemase-producing Enterobacteriaceae or meticillin-resistant S. aureus were initially treated with inactive drugs. Overall, 36.3% of patients could have been treated with appropriate therapy at least 24 h earlier if molecular data had been used.

Tailored cytomegalovirus management in lung transplant recipient: a single-center experience.

INTRODUCTION: Among solid organ recipients lung transplant recipients are at highest risk to be affected by cytomegalovirus infection (CMV) or to die from CMV disease. Two strategies are usually adopted in the clinical management of transplant recipients: antiviral prophylaxis and pre-emptive therapy. METHODS: In our center we adopted from 2007 a combined prophylaxis with anti-CMV immunoglobulins in the first post-transplant year and antiviral therapy (gancyclovir or valgancyclovir) from post-transplant day 15 for 3 weeks and in case of CMV bronchoalveolar lavage specimen positivity (polymerase chain reaction or shell vial). Moreover, we studied specific cellular immune response by an Elispot assay to define responder patients by the number of spot forming units (<5 nonresponders, 5-20 weeks, 20-100 good, >100 very good responders). RESULTS: We reduced acute rejections (from 17% to 6%, odds ratio 3.25), lymphocytic bronchitis bronchiolitis (from 11% to 2%), and first-year CMV pneumonia after the first post-transplant month (from 6.4% to 1%). We showed in nonresponders an earlier onset (68 vs 204 post-transplant days) and a longer duration (>14 days vs <14 days) of infection (P < .05 for all referred data). DISCUSSION: The morbility reduction has been obtained by antiviral therapy, increasing costs and risk of side effects. Our more recent studies show a population with a good immune response that probably doesn't need a pharmacological intervention but just a strict follow-up. CONCLUSION: Our proposed strategy is now tailoring the therapy on immune response clinical application, limiting to the specimen positivity in nonresponders.

Evaluation of Epstein-Barr virus-specific immunologic response in solid organ transplant recipients with an enzyme-linked ImmunoSpot assay.

Epstein-Barr virus (EBV) is a γ-herpes virus, responsible for infectious mononucleosis in immunocompetent hosts. Cellular immunity appears rapidly during EBV primary infection, keeping it silent despite long-life persistence in B lymphocytes. Defects of the EBV-specific cellular immunity are supposed to be the basis of post-transplantation lymphoproliferative disorders, promoted by high levels of immunosuppression. We retrospectively reviewed 197 solid organ transplant recipients to investigate EBV-specific lymphocyte responsiveness using Enzyme-linked ImmunoSpot assay (EliSpot), which assesses the EBV-specific interferon (IFN)-γ producing peripheral blood mononuclear cells, and kinetics of EBV infection/reactivation post-transplantation using quantitative real-time polymerase chain reaction (PCR) on whole blood. Overall, 102 of the 197 patients (51.8%) showed EBV responsiveness at the EBV-EliSpot assay: 68 (66.6%) showed a persistently positive EBV response in 3 or more determinations and 34 (33.3%) had transient episodes of nonresponsiveness. Ninety-five (48.2%) patients were persistently EBV nonresponders. EBV-DNAemia data were available for 58 patients: 27.6% presented at least one episode of EBV-DNA occurrence. No differences were found in EBV-EliSpot response stratification between the groups of patients who experienced episodes of EBV reactivation and those without EBV-DNAemia. However, EBV DNAemia peak values tended to be higher in the first year post-transplantation in the group of patients with a persistent positive EBV-specific immune response. EBV viral load quantitation in blood and EliSpot EBV-specific immune response determination may represent a powerful tool for monitoring solid organ transplant recipients, guiding immunosuppression modulation in patients with active EBV replication.

Clinical impact of HSV-1 detection in the lower respiratory tract from hospitalized adult patients.

The occurrence and clinical impact of herpes simplex virus (HSV) were evaluated in 342 bronchoalveolar lavage specimens from 237 patients. HSV-1 and HSV-2 were detected in 32.1% and <1% of patients, respectively. A significant difference of HSV-1 prevalence and load was found in relation to admission to intensive care unit, mechanical ventilation and mortality within 28 days; in particular, a viral load ≥10(5) copies/mL bronchoalveolar lavage fluid was significantly associated with critical features. No association was found with immune status or other characteristics. Nine of 21 (42.9%) cases of ventilator-associated pneumonia were positive for HSV-1, with poor outcome in six.

The lack of cytomegalovirus-specific cellular immune response may contribute to the onset of organ infection and disease in lung transplant recipients.

Cellular immune response has been demonstrated to play a role in the control of human cytomegalovirus (HCMV) replication in organ transplant recipients. Herein, HCMV-specific T-cell response and association to the onset of organ infection/disease were prospectively evaluated by EliSPOT assay in a population of 46 lung transplant (LT) recipients at 1, 3, 6, 9 and 12 months post-transplantation. According to our centre?s practice, a combined prolonged antiviral prophylaxis (HCMV-IG for 12 months and ganciclovir or valganciclovir for 3 weeks from postoperative day 21) was given to all LT recipients. HCMV-DNA was concomitantly detected on bronchoalveolar lavage (BAL) and whole blood by real-time PCR. Approximately one third of patients resulted HCMV persistently non-responder; the rate of HCMV infection, as evaluated by HCMV-DNA positivity, tended to be higher in non-responders. Mean viral load on BAL was significantly higher in non-responders vs other patients (p < 0.001). Temporal profile of infections appeared related to the HCMV responder status with a shorter time to onset of infection post-transplantation and a longer duration in non-responders. The occurrence of organ disease (i.e. pneumonia) tended to be higher in non-responders, with poor prognosis, as death occurred in one of three non-responder patients that developed HCMV pneumonia. The lack of HCMV-specific cellular response can contribute to the onset of organ infection and disease also in patients in which antiviral prophylaxis was adopted; this could be due to the potential occurrence of incomplete control of replication in lungs or a delayed priming of T-cell reconstitution.

Combined measurement of serum DNA and urine VP1 messenger RNA in monitoring BK virus replication in kidney graft recipients.

Evaluation of BK virus replication is a fundamental tool in the monitoring of renal transplant recipients. Herein, we investigated the role of urine VP1 messenger RNA (mRNA) quantification and combined measurement of serum DNA and urine VP1 mRNA in 428 kidney allograft recipients. BK viremia and viruria were detected in 24 (5.6%) and 54 (12.6%) patients, respectively. A diagnosis of BKV-associated nephropathy (BKVAN) was established in 2 patients, both within the first year posttransplantation. Based on urine VP1 mRNA measurement, BKV replication was observed in 10 (2.1%) patients, 2 of whom displayed BKVAN. Urine VP1 mRNA was detected in all cases in association with viremia except 5 and in all cases with viruria. No difference among VP1 mRNA levels was noted between the 2 BKVAN patients and the highest values in patients without BKVAN. The urine VP1 mRNA result by analysis using the operating characteristics was not superior to viremia, despite the improvement obtained with the combined measurement of viremia (cut-off, 16,000 copies/mL) and urine VP1 mRNA (>10,000 copies/10(3) cells). In conclusion, VP1 mRNA measurements may complement viremia and viruria to monitor BKV replication, although its use is limited by its technical complexity in comparison with DNA detection.

Evaluation and significance of cytomegalovirus-specific cellular immune response in lung transplant recipients.

In lung transplant recipients, cytomegalovirus (CMV) has been associated with direct ie, organ and systemic infection/disease, and indirect effects, including predisposition to develop acute rejection episodes and chronic allograft dysfunction. Cellular immune responses have been demonstrated to play a role in the control of CMV replication. We evaluated CMV-specific cellular responses among lung transplant recipients associated with the onset of organ infection/disease. Cellular responses were evaluated by an Elispot assay of 48 specimens from 24 patients. All samples were evaluated beyond 1 year after transplantation; CMV DNA was concomitantly detected in bronchoalveolar lavage (BAL) and whole blood specimens. Each patient received a combined prolonged antiviral prophylaxis with CMV Ig for 12 months and gancyclovir or valgancyclovir for 3 weeks after postoperative day 21. Nine patients (37.5%) showed transient or persistent CMV nonresponses including donor-recipient negative serologic matching in 2 cases. Positive CMV DNA results were observed in 18/48 BAL specimens (37.5%) from 12 patients (50%). A viral load of >10(4) copies/mL was observed in only 3 cases, 2 of whom were positive also on whole blood. Among these 3 patients, 2 were responders and BAL (as well as whole blood) specimens collected subsequently were negative for CMV DNA; 1 nonresponder patient exhibited a viral load of 426,492 copies/mL BAL (DNAemia, <2,000 copies/mL), developed CMV pneumonia (confirmed by histopathology and immunohistochemistry) and died within 28 days. The prevalence of CMV DNA positivity on BAL did not differ in relation to the immune response; the mean viral load on BAL showed significantly higher results among nonresponders than responders, namely, 1.4 × 10(5) ± 2.4 × 10(5) copies/ml versus 7.9 × 10(3) ± 1.4 × 10(4) (P=.02). Evaluation of CMV-specific cellular immune responses by in vitro immunologic monitoring complements virologic monitoring, helping to identify lung transplant recipients at risk of developing organ infection/disease.

Detection of the novel variant of influenza virus A/H1N1v in bronchoalveolar lavage of adult hospitalized patients during the 2009/2010 winter season.

AIM: The aim of this study was to report most recent data regarding the occurrence of influenza A virus H1N1v in the lower respiratory tract from a cohort of hospitalized adult patients during the winter season 2009/2010 and investigated the main clinical features and outcomes. METHODS: A total of 130 consecutive BAL specimens (collected from October 2009-March 2010) of 101 patients were retrospectively analyzed for influenza A virus H1N1v positivity using a commercial kit. RESULTS: Overall, 19/130 (14.6%) BAL specimens from 17/101 (16.8%) patients were positive for the novel influenza A H1N1v virus. H1N1v resulted significantly more prevalent in immunocompetent subjects. As regards clinical features, H1N1v resulted more prevalent in respiratory insufficiency or acute respiratory illness. Thirteen patients died during the analytic period; three of them (23.1%) resulted positive to H1N1v but no direct association has been made. CONCLUSION: Our cohort study of influenza A H1N1v detection in BAL from hospitalized adult patients confirms the overall moderate clinical impact of this virus, as reported in most reports worldwide. It remains to be evaluated the role of reassortment with influenza virus strains circulating in the winter season 2010/2011 and its potential pathogenicity.

Human herpesvirus 7 detection by quantitative real time polymerase chain reaction in primary cutaneous T-cell lymphomas and healthy subjects: lack of a pathogenic role.

BACKGROUND: Primary cutaneous T-cell lymphomas (CTCLs) are a heterogeneous group of lymphomas where the tumour population emerges within a multiple subclone pattern. Mycosis fungoides (MF) and Sézary syndrome (SS) are characterized by the expansion of clonal CD4+/CD45RO+ memory T cells. Lymphomatoid papulosis (LyP) is a chronic, lymphoproliferative disorder included in the CD30+ primary CTCL spectrum. Several studies have suggested a role of viral infection for super-antigenic activation of T lymphocytes; however, evidence of their association with CTCLs is still lacking. Human herpesvirus (HHV) 7 is a CD4+ T-lymphotropic herpesvirus; its restricted cellular tropism and the ability to induce cytokine production in infected cells could make it an important pathogenic cofactor in lymphoproliferative disorders. OBJECTIVES: To investigate the presence of HHV7 DNA on CTCL and healthy skin donors (HD). METHODS: We used quantitative real time polymerase chain reaction to evaluate the potential pathogenic role of HHV7. RESULTS: Twenty-seven of 84 (32.1%) HD were positive for HHV7 DNA. Twenty-one of 148 (14.2%) patients with CTCLs were positive for HHV7 DNA: nine of 39 (23.1%) SS, six of 14 (42.9%) CD30+ CTCLs and six of 24 (25.0%) LyP, and HHV7 DNA was negative in all 71 patients with MF. CONCLUSIONS: These results seem to exclude a pathogenic role of HHV7 in CTCLs, suggesting the possibility of skin as a latency site.

Monitoring of human Cytomegalovirus infection by antigenemia and viremia in the first 100 days following renal transplantation and relation to antiviral strategies.

AIM: Human Cytomegalovirus (HCMV) is a relevant pathogen in transplant recipients, particularly in the first three months post-transplantation. The use of antiviral prophylaxis and pre-emptive therapy is able to reduce incidence of HCMV infection and disease. The incidence of HCMV infection and disease in renal transplant recipients in the first 100 days post-transplantation was investigated, in relation with HCMV serological matching and therapeutic management. METHODS: Incidence of HCMV infection in the first 100 days post-transplantation was evaluated by pp65-antigenemia in 165 patients on a total number of 1241 clinical samples. Patients were divided in four groups according to donor/recipient serological matching: D(-)/R(-) (low risk of HCMV disease), D(-)/R+ and D+/R+ (intermediate risk) and D+/R(-) (high risk). Antiviral strategy (prophylaxis in high risk group; pre-emptive therapy in intermediate risk group, no therapy in low risk group) and immunosuppressive protocol were recorded. RESULTS: Incidence of antigenemia-positivity was as follows: 0/3 D(-)/R(-) patients; 59/130 (45.4%) D+/R+; 5/16 (31.3%) D(-)/R+; 4/16 D+/R(-). No significative difference was found between the four groups in terms of incidence of antigenemia-positivity in the first 100 days following transplantation. Antigenemia values >50 pp65-positive/2x10(5) peripheral blood leukocytes (used to start pre-emptive therapy) were present in 18/130 (13.8%) D+/R+; 1/16 (6.2%) D+/R(-); 0/16 D(-)/R+. Viral kinetics in patients with HCMV infection was described. CONCLUSION: No significative difference was found in terms of incidence of HCMV infection in the first 100 days post-transplantation in relation to immunosuppressive protocol and serological matching, suggesting the appropriateness of antiviral strategies and viral monitoring adopted in this setting.

Laparo-endoscopic "rendezvous": a new technique in the choledocholithiasis treatment.

BACKGROUND/AIMS: Endoscopic sphincterotomy for common bile duct stone clearance during laparoscopic cholecystectomy may fail due to difficulties in cannulating the papilla major. In this study we propose a new technique that facilitates the cannulation of the papilla and the common bile duct stone clearance during a standard laparoscopic cholecystectomy. Its clearance percentage, complication rate and post-operative stay have been evaluated and compared with standardized procedures such as open surgery and endoscopic sphincterotomy before laparoscopic cholecystectomy. METHODOLOGY: In a group of 16 patients presenting with cholelithiasis and common bile duct stones or papillitis, the sphincterotome was driven across the papilla into the choledochus by a Dormia basket passed in the duodenum through the cystic duct during laparoscopic cholecystectomy. Measures of outcome were clearance rate, mortality, morbidity and hospital stay. Furthermore, data obtained from this sample of patients were compared with those from another two groups of 16 patients in which choledocholithiasis was managed either by endoscopic sphincterotomy performed before laparoscopic cholecystectomy or by open cholecystectomy and trans-duodenal sphincterotomy. RESULTS: The rate of cannulation of the papilla and of the common bile duct stone clearance was 100% when the combined endo-laparoscopic approach was used in 15 patients with endoscopic sphincterotomy (93,7%) and in 15 patients with open sphincterotomy (93,7%), cholecystectomy was successful in every case. The groups were statistically similar with regard to complications; none of the patients required blood transfusion. The mean post operative stay was 95.2 hours (range 48-240) for the first group, 350.1 hours (range 192-1680) for the second and 69.7 hours (range 24-132) for the third. CONCLUSION: The laparo-endoscopic rendezvous, though still in evolution, is an efficacious method which can be used during the laparoscopic strategy of common bile duct clearance.

[Prognostic risk factors in patients operated on for perforated peptic ulcer. A retrospective analysis of critical factors of mortality and morbidity in a series of 40 patients who underwent simple closure surgery]. / Fattori di rischio prognostico in pazienti operati per ulcera peptica perforata. Analisi retrospettiva dei fattori critici per la mortalità e morbilità su una serie di 40 pazienti sottoposti ad intervento di sutura semplice.

STUDY OBJECTIVE: To identify factors affecting mortality and morbidity in patients operated on for perforated peptic ulcer. DESIGN: Retrospective analysis. SETTING: University Hospital, Italy. PATIENTS: Forty patients consecutively operated on for perforated peptic ulcer by simple suture procedure performed either by laparotomy (n = 26) or laparoscopic (n = 14) approach. MEASUREMENTS AND MAIN RESULTS: Mortality was 20% (n = 8) and morbidity in survivors was 25% (n = 8). Compared to survivors, non-survivors were older (mean age 79.3 yrs. vs 60.0 yrs., p < 0.01), had worse APACHE II and SAPS scores (mean 20.1 vs 8.5, p < 0.001; and 13.1 vs. 5.5, p < 0.0001 respectively), were treated later (mean interval from outbreak of symptoms to surgery 30.8 hrs. vs. 11.1 hrs., p < 0.01), and the size of their perforation was larger (mean 15.1 mm. vs. 8.6 mm, p < 0.05). The laparoscopic approach was the only factor that significantly was associated with morbidity in survivors (p < 0.01). The presence of at least two risk factors, enhanced the probability of death. CONCLUSION: Old age, great APACHE II and SAPS scores, delay in treatment and large size of the perforation were associated significantly to mortality in perforated peptic ulcer patients. Efforts should be made perioperatively for patients having these risk factors.

Single and double stapled anastomoses in rectal cancer surgery; a retrospective study on the safety of the technique and its indication.

Low anterior resection is commonly believed the main indication to double stapled (DS) technique, because placing the purse-string suture on the distal rectum is difficult or impossible. This study was designed to figure out the safety of the DS technique and to better define its role in rectal cancer surgery. The data of 34 patients that had a DS anastomosis were retrospectively compared to those of 43 that had a single-stapled (SS) anastomosis after anterior resection. Three deaths after SS (7%) and one after DS procedures (3%) were recorded (p = 0.62). Rates of clinical leaks were 12% (four cases) in the DS group and 14% (six cases) in the SS group (p = 0.41). The mean distance of the rectal tumour from the anal verge was significantly lower for DS (mean = 7.7 cm) respect to SS (mean = 12.7 cm) anastomoses (p < 0.0001) and the blood consumption at surgery was significantly greater in patients that had DS (mean = 375 ml) compared to SS-anastomoses (mean = 180 ml) (p = 0.028). Thus, the DS technique was mostly used in patients at high risk for leakage. The study shows that DS technique is a safe and reliable method to perform colorectal anastomosis after anterior resection for cancer. For cancers located in the upper rectum the routine adoption of the DS increases the cost of surgery and does not offer advantages over the SS technique with the exception of making feasible end-to-end mechanical anastomoses involving bowel segments having different diameters.

Omeprazole versus ranitidine plus somatostatin in the treatment of severe gastroduodenal bleeding: a prospective, randomized, controlled trial.

In a randomized, controlled clinical trial omeprazole was compared with ranitidine plus somatostatin in the treatment of severe acute gastrointestinal bleeding due to peptic pathology. Intravenous infusion of the drugs was administered until clinical stabilization or surgical operation. The two regimens were equally effective in controlling bleeding. The need for blood transfusion and surgical operation together with the mortality rate did not differ significantly between groups. No toxic effects were observed as a result of the infusion of omeprazole. In this study the infusion of omeprazole alone showed an efficacy comparable to the association of ranitidine and somatostatin in the treatment of severe acute peptic bleeding.

[Ultrasonic diagnosis of gastroduodenal perforation]. / Diagnostica ecografica di perforazione gastro-duodenale.

Ultrasonography (US) is routinely used as a diagnostic approach in the surgical acute abdomen, even though its major limitation is the demonstration of the hollow viscus. In the present paper, the results obtained with US in 12 cases of gastric or duodenal perforations are reported. The diagnosis, of gastroduodenal perforation was correctly made in 58% of cases by means of US; in 91.7% of cases, at least one US finding correlable with gastroduodenal perforation was observed. Even though US exhibits poorer diagnostic sensitivity than conventional radiology, it can be considered a valuable diagnostic tool in the early diagnosis of gastroduodenal perforation especially when radiographic findings are negative.

Subcutaneous seeding after percutaneous ethanol injection of liver metastasis.

A 72-year-old cirrhotic woman underwent percutaneous ethanol injection treatment of a liver metastasis of unknown origin. A subcutaneous metastasis developed at the site of the punctures. Needle track seeding is a rare complication of fine-needle biopsy but has never--to the authors' knowledge--been reported after percutaneous ethanol injection. The possible causes of this complication are discussed.