Angela Di Matteo

Nuclear expression of the lower matrix protein of human cytomegalovirus in peripheral blood leukocytes of immunocompromised viraemic patients

Peripheral blood leukocytes (PBL), namely polymorphonuclear leukocytes (PMNL), are the major carrier of human cytomegalovirus (HCMV) in the blood of immunocompromised patients with HCMV viraemia. By using monoclonal antibodies (MAbs) directed against different early and late viral proteins, we showed that the protein accumulating in PBL, originally reported to be an immediate early (IE) gene product, is the 65K lower matrix early structural protein (ep65). This protein is detectable by immunofluorescence before IE proteins during early stages of the replication cycle of HCMV in permissive human embryonic lung fibroblast cells. However, the appearance of ep65 in the nucleus within 1 h post-infection in the presence of cycloheximide indicates that it represents uptake from the virus inoculum rather than newly synthetized protein. The ep65 MAbs staining PBL did not react with Vero cells infected with a recombinant vaccinia virus encoding the major IE gene (IE1) product, whereas MAbs reactive with the 72K major IE protein stained only faintly a small number of infected PBL. A group of four ep65 MAbs was tested in competitive binding assays to show that ep65 possesses at least three distinct epitopes. These were recognized by all four MAbs in AD169-infected Vero cell cultures when fixed with formaldehyde, whereas only one MAb recognizing a distinct epitope was reactive with methanol-acetone (MA)-fixed AD169-infected Vero cells. In formalin-fixed PBL the number of infected cells stained by the four ep65 MAbs was about twofold that found using MA-fixed cells. Using fluorescence-activated cell sorter-purified leukocyte subpopulations from viraemic patients with different levels of viraemia, the ratio of ep65-positive to ep65-negative cells was found to be 1:100 to 1:100,000 for PMNL, and only 1:10,000 to 1:100,000 for mononuclear cells.

Effect of exercise and strength training on cardiovascular status in HIV-infected patients receiving highly active antiretroviral therapy

A routine evaluation of lipid metabolism and body fat distribution along with a careful cardiovascular risk stratification according to international guidelines are required for HIV-infected patients receiving highly active antiretroviral therapy. Intervention includes evaluation of patients for both epidemiological and clinical factors, and for anthropometric and biochemical parameters. Diet counseling, prescription of antihyperlipidemic drugs and exercise training are the cornerstones of programs devoted to protecting patients from side effects of therapies that compromise quality of life and the functions of organs like the pancreas and heart that are involved in lipid disorders.

Clinical and therapeutical follow-up of HIV-associated pulmonary hypertension: prospective study of 10 patients.

Objective: To determine the clinical course and prognosis of pulmonary hypertension (PH) in HIV-infected patients in comparison with a group of PH patients without HIV infection. The secondary objective was to ascertain whether more powerful antiretroviral treatments (highly active antiretroviral therapy) could modify the course of PH in HIV-infected patients. Design: Patients with PH and HIV (HIV-PH, group 1) and patients without HIV (PPH, group 2) were prospectively followed. Setting: A tertiary care institution. Patients: Group 1 included 10 patients, and group 2 included 25 patients. Interventions: In group 1, HIV infection was staged according to Centers for Disease Control and Prevention (CDC) classification when patients entered the study, and was re-staged every fourth month. In both groups, PH functional classes and right heart catheterization (RHC) were determined at baseline. Results: In group 1, one of 10 patients was assigned to New York Heart Association (NYHA) class II, seven patients to NYHA class III, and two patients to NYHA class IV. CDC stages ranged from A1 (three patients) to C3 (one patient). No patient showed progression of HIV disease during follow-up. By May 2001, six patients had died. The median survival by the time of RHC was 15.1 months. Causes of death were heart failure in three cases, sepsis in two, and suicide in one case. In seven patients, epoprostenol was started; three patients survived and four died. The cause of death was heart failure in one patient, suicide in one, non-catheter-related sepsis in one patient and catheter-related sepsis in the last patient. In group 2, 11 patients out of 25 were assigned to NYHA class II, 11 patients to NYHA class III, and three patients to NYHA class IV. RHC was not statistically different in the two groups. By May 2001, nine of 25 patients died and one underwent a double-lung transplant. The median survival from the time of RHC was 6.86 months. Cumulative survival rates by RHC were not statistically different (hazard ratio close to 1). Conclusions: In HIV-infected patients, the onset of PH adversely affects the prognosis at any stage of infection. Clinically adverse progression of PH is not correlated with HIV initial stage and evolution. Moreover, prognosis in patients with sporadic or familial PPH and in patients with HIV-PH with similar RHC is so similar as to strengthen the concept that pulmonary vascular disease overshadows the overall clinical problem in HIV-infected patients.

Human Enteric Coronaviruses: Further Characterization and Immunoblotting of Viral Proteins

1. Myer EA, Jarroll EL. Giardiasis. Am J Epidemiol 1980;111:1-12 2. Keystone JS, Krajden S, Warren MR. Person-to-person transmission of Giardia lamblia in day-care nurseries. Can Med Assoc J 1978;119:241-8 3. Jarroll EL, Bingham AK, Meyer EA. Effect of chlorine on Giardia lamblia cyst viability. Appl Environ Microbiol 1981;41:483-7 4. Owen RL. The immune response in clinical and experimental giardiasis. Trans R Soc Trop Med Hyg 1980;74:443-5 5. Smith PD, Gillin FD, Brown WR, Nash TE. IgG antibody to Giardia lamblia detected by enzyme-linked immunosorbent assay. Gastroenterology 1981;80:1476-80 6. Myer EA. Giardia lamblia isolation and axenic cultivation. Exp Parasitol 1976;39:101-5 7. Smith PD, Gillin FD, Kausha Na, Nash TE. Antigenic analysis of Giardia lamblia from Afganistan, Puerto Rico, Ecuador, and Oregon. Infect Immun 1982;36:714-9 Concise Communications

Pulmonary Cladophialophora boppii Infection in a Lung Transplant Recipient: Case Report and Literature Review

Cladophialophora boppii is a dematiaceous fungus, which has been reported only rarely to be the cause of cutaneous infection. Herein we describe a C boppii parenchymal and bronchial infection in a lung transplant recipient. We also illustrate the clinicoradiologic patterns and review possible treatment options for these difficult infections.

Rapid detection of human rotavirus strains in stools by single-sandwich enzyme-linked immunosorbent assay systems using monoclonal antibodies

Using murine monoclonal antibodies (MAbs) raised against the common antigen of group A rotavirus (RV), two single-sandwich ELISA systems were developed for detection of RV in stools: one using polyclonal antibody (PAb) as capture and a MAb as detector antibody (referred to as PAb-MAb assay); and the other based on the use of two different MAbs as capture and detector antibodies (referred to as MAb-MAb assay). In each single-sandwich ELISA system, samples and peroxidase-labeled MAb were incubated sequentially (two-step method) or simultaneously (one-step method). Using the two-step procedure on purified RV, 50 pg of protein was detected in the PAb-MAb as well as in the MAb-MAb assay, whereas the one-step method detected 0.4 ng and a conventional double-sandwich ELISA detected 3.2 ng of viral protein. Titration of RV samples from stools and cell cultures showed that single-sandwich ELISA titers were, on the average, 10-100-fold higher than those obtained by electron microscopy (EM), but 10-100-fold lower than those obtained by solid-phase immune EM (SPIEM). However, when 200 stool samples previously examined by EM or SPIEM were tested by the single-sandwich ELISA systems, specificity and sensitivity of these assays were 100%, and comparable to SPIEM. No false positive results were obtained when 54 samples of meconium and 91 stools from newborns in the first five days of life were tested. The two-step procedure appeared to be somewhat preferable over the one-step method, which, although faster, gave a marked prozone with a few samples in the MAb-MAb assay. The use of MAbs in rapid single-sandwich ELISA systems for RV detection in stools appears highly convenient, due to reliable results and short test performance times.

Characterization of rotavirus subgroup-specific monoclonal antibodies and use in single-sandwich ELISA systems for rapid subgrouping of human strains

SummaryTwo subgroup-specific monoclonal antibodies (MAb) raised in mice against group A human rotavirus were shown to react by immunoblotting with the trimeric form of VP6 of the homologous subgroup and successfully applied to development of new single-sandwich ELISA systems for rapid subgrouping of human strains. All of the 344 strains tested could be subgrouped, but for two of them prior propagation in cell cultures was required.

HIV Infection-Related Cachexia and Lipodystrophy

Protein energy malnutrition (PEM) is, alone or associated with other diseases, the first step in the development of cachexia [1] [3]. An insufficient amount of food is the leading cause of malnutrition and infectious diseases are the second. In developing countries, 20% of the population — more than 800 million people — eats a quantity of food only sufficient to supply energy for a sedentary life, i.e. 1.2–1.4 times the resting energy expenditure (REE).More than 192 million children suffer from PEM and 2 billion people lack different micronutrients (vitamins, minerals, essential fatty and amino-acids) [4] [7]. Even in Western countries, where an enormous surplus of food is produced, many groups of people, especially the poor, the elderly, drug addicts, pregnant women, patients with liver, kidney and gastro-intestinal (GI)-tract diseases, cancer, AIDS, show nutritional defects. In general, 60% of the world’s population (41% in developing countries) consumes less than 2600 Kcal/person/day, an amount of energy considered barely sufficient for limited activity.

Nutritional Status Assessment

Malnutrition is not invariably diagnosed by physical findings of nutritional deficits. Malnutrition is a deviation (in excess or defect) from a complex of ideal scores. Paradoxically, a plump, flourishing patient may be affected by malnutrition if he or she exceeds this ideal score.

Neurological Complications Involving the Central Nervous System After Allogeneic Hematopoietic Stem Cell Transplantation During a Period of Evolution in Transplant Modalities: A Cohort Analysis.

Background Neurological complications (NC) after hematopoietic stem cell transplantation (HSCT) are rare events. The evolution of transplant procedures has resulted in improved survival and has allowed elderly patients or those with comorbidity to receive an HSCT. The risk of NC in these patients has still not been well defined. Therefore, we carried out an observational study to estimate the occurrence and identify the risks associated with NC. Methods The study cohort included 452 adult-allogeneic HSCT recipients, transplanted from 1997 to 2012. The median follow up was 1.3 year (0-15.7). A myeloablative regimen was used in 307 patients. Two hundred patients were grafted from matched unrelated donor (MUD), of these, 129 (64.5%) received an in vivo T-cell depletion. Results Out of 452 patients, 30 (6.6%) developed NC. Infections were the most frequent causes of NC (30%). Overall survival decreased in patients developing NC (P < 0.001). Univariate survival regression on the cumulative incidence of NC identified period of transplant, linear trend between 4-year periods (1997-2012) (P < 0.001), MUD (P < 0.001), and recipients age (P = 0.034) as significant risk factors. In multivariate analysis, period of transplant (P < 0.001) and MUD (P = 0.004) remained significant independent risk factors. Matched unrelated donor recipients showed a 3.8-fold elevated risk of developing NC. Conclusions Analysis highlights a temporal trend of incidence of NC that progressively increased over time and confirms a strong association between donor type and risk of NC. Our observations suggest that, although relatively uncommon, NC after allo-HSCT, may become more frequent due to the improved overall survival in recent years.