Search PubMed⌕ Search

Biomedical subjects

R Garofalo

Publications and source records attributed to R Garofalo.

53 records · Page 3Linked to original sources

Preparation of respiratory syncytial virus subgroup A and B antigens for enzyme immunoassay antibody detection.

A simplified method was described for purification of respiratory syncytial virus (RSV) subgroup A and B aimed to be used as antigens in enzyme immunoassay (EIA). The titer of each RSV subgroup and the amount of protein was determined from the visible band in 45% sucrose gradient. The quality of prepared RSV subgroup antigens for EIA was described in terms of the achievable final titer, the amount of protein, and EIA criss-cross titration. The RSV subgroup A and B antigens, diluted as 1:100 (low opalescent band in 45% sucrose layer) or 1:800 (high opalescent band in 45% sucrose layer) produced a positive reaction in EIA criss-cross titration with IgG antibodies from the patient's serum (convalescent phase) diluted as 1:25,600 (for RSV A) and 1:6,400 (for RSV B). This method offers shorter and more simplified steps of viral antigen purification, and provides acceptable quantity and quality of viral antigens appropriate for use in EIA.

Antibodies, Viral↗

Presence and biological activity of a GnRH-like factor in the nervous system of Helisoma trivolvis.

Gonadotropin-releasing hormone (GnRH) constitutes a family of neuropeptides found throughout the vertebrates. Although a GnRH-like peptide has also been isolated from yeast (alpha-mating factor), the presence of GnRH has not been clearly demonstrated in invertebrate phyla. In this study, we tested the hypothesis that GnRH-like peptides are present and functional in the central nervous system (CNS) of the gastropod mollusc, Helisoma trivolvis. The presence of a GnRH-like peptide was examined by three methods: (1) in immunofluorescence studies with four different antibodies generated against several GnRH peptides, select neurons and putative neurosecretory cells were specifically and consistently labelled throughout the CNS; (2) reverse-phase high performance liquid chromatography (HPLC) and radioimmunoassay (RIA) analysis revealed a GnRH-like factor which co-migrates with mammalian (m)GnRH; and (3) in bioactivity experiments, extracts of Helisoma trivolvis CNS mimicked GnRH in stimulating gonadotropin release from dispersed goldfish pituitary cells in static culture. Two functional assays were carried out to examine the potential biological roles of GnRH-like peptides in Helisoma. (1) Intracellular recordings of left-parietal and visceral ganglion neurons revealed diverse electrophysiological responses to mGnRH. These effects were attenuated by a mGnRH antagonist. (2) Addition of mGnRH arrested neurite outgrowth in a subpopulation of dissociated embryonic Helisoma neurons in culture. Taken together, these results strongly suggest that a mGnRH-like peptide is an important neuropeptide in Helisoma. A hypothesis is presented that GnRH-like peptides may be ancient factors that are conserved both structurally and functionally in the evolution of animals.

Animals↗

Radical tumor excision and cosmetic balance in the surgical treatment of breast carcinoma: biquadrantectomy.

The present trend in favor of conservative surgery is in contrast with histopathological findings of multicentric breast carcinoma, which may be responsible for the occurrence of relapses in spite of the use of radiation therapy. As a consequence, conservative surgery appears to be inadequate, especially when tumor size exceeds 2 cm. Therefore for tumors up to 4 cm in size, which are not adherent to the pectoral fascia, we propose the excision of two quadrants with concomitant operation on the contralateral breast resulting in a more symmetrical and therefore cosmetic effect. This is a more radical procedure than the removal of the one quadrant.

Breast Neoplasms↗

Eosinophil degranulation in the respiratory tract during naturally acquired respiratory syncytial virus infection.

Eosinophil cationic protein (ECP), a cytotoxic protein contained in the granules of eosinophils, has been suggested as having an important role in the pathogenesis of asthma. To determine whether ECP plays a similar role in bronchiolitis, we tested samples of nasopharyngeal secretions, obtained from a group of 47 children with various forms of illness related to respiratory syncytial virus (RSV) and from 26 children with non-RSV upper respiratory tract illness or bacterial pneumonia, for the presence of ECP by means of a double-antibody radioimmunoassay. Concentrations of ECP in children with RSV bronchiolitis were significantly higher (166.8 ng/ml) than the mean concentration of ECP in both groups of children with RSV upper respiratory tract illness (43.5 ng/ml, p less than 0.002) and RSV lower respiratory tract disease without wheezing (29.1 ng/ml; p less than 0.0002). Children with non-RSV upper respiratory tract illness or bacterial pneumonia had levels of ECP in nasopharyngeal secretions similar to those of children with RSV upper respiratory tract illness or RSV pneumonia. High ECP levels in nasopharyngeal secretions (greater than 50 ng/ml) were predictive of the development of bronchiolitis at the time of RSV infection (p less than 0.001), and the individual ECP levels correlated with severity of the disease as determined by the initial PaO2 concentrations (p less than 0.05). These data suggest that eosinophil degranulation in the respiratory tract occurs during RSV bronchiolitis and may play a significant role in the development of virus-induced airway obstruction.

Bacterial Infections↗

Activation of human eosinophils in vitro by respiratory syncytial virus.

To determine the nature of the interaction between viruses and eosinophils, normodense eosinophils were separated from the blood of healthy volunteers and incubated in vitro with respiratory syncytial virus (RSV). After incubation for 2 h with the virus, 29.5 +/- 15.8% of the eosinophils demonstrated specific binding of the virus to the cell membrane, as detected by fluorescent staining with an anti-RSV MAb. Superoxide production and leukotriene C4 release were measured as determinants of cell activation. Using a cytochrome c reduction assay, superoxide could be detected in the supernatant 30 min after exposure to RSV. Maximal release was reached at 3 h postexposure (5.88 +/- 2.19 nmol cytochrome c reduction/5 x 10(5) cells). The virus-induced superoxide generation varied in magnitude among different subjects and ranged from 0.6 to 11.5 nmol cytochrome c reduction/5 x 10(5) cells. RSV also appeared to prime eosinophils to the effects of other known cell activators, as demonstrated by an increase in superoxide production upon subsequent stimulation of RSV-primed cells with phorbol-12-myristate-13-acetate (21.4 +/- 5.8 versus 9.4 +/- 2.7 nmol cytochrome c reduction/5 x 10(5) cells for primed and unprimed cells, respectively) (p less than 0.04). RSV did not directly induce leukotriene C4 release from the eosinophils but primed the cells to exhibit a more vigorous response on subsequent challenge with the calcium ionophore A23187 (9.16 +/- 1.04 versus 4.2 +/- 1.3 ng leukotriene C4/1 x 10(6) cells) (p less than 0.005). These findings indicate that RSV can activate or prime eosinophils to release various inflammatory mediators.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Release of leukotriene B4 from human neutrophils after interaction with nontypeable Haemophilus influenzae.

Opsonization of nontypeable Haemophilus influenzae with antibody is critical for the interaction between the organism and human polymorphonuclear leukocytes (PMNs). Nontypeable H. influenzae opsonized in fresh antibody-positive serum induced the release of 42.5 +/- 17.9 ng of leukotriene B4 per ml from PMNs after 20 min of incubation at 37 degrees C. On the other hand, opsonization of the organisms in fresh antibody-negative serum stimulated the release of significantly smaller amounts of leukotriene B4 by the PMNs. Simultaneous determinations of phagocytosis demonstrated similar patterns of response. A small amount (26.7 +/- 7.6%) of unopsonized nontypeable H. influenzae was phagocytosed by PMNs during 20 min of incubation at 37 degrees C. In contrast, 89.3 +/- 2.0% of nontypeable H. influenzae opsonized in fresh antibody-positive serum was phagocytosed during the same incubation period (P less than 0.001). Removal of complement through heat inactivation at 56 degrees C for 30 min did not significantly affect phagocytosis. These data suggest that the humoral immune response to nontypeable H. influenzae plays an important role in the inflammatory process and may contribute to the production of middle ear effusions in otitis media.

Adult↗

[Role of the ultrasonic examination in the diagnosis of pelvic pathology].

The diagnostic accuracy of sonography in pelvic pathology is assessed in the case in which the operator is aware of the clinical report. Results attribute in absolute terms more effectiveness to sonography than to the clinical examination but it must, in any case, be considered as an extension of the gynaecological examination and not a replacement of the anamnestic history and physical examination.

Diagnostic Errors↗

Lumbar disc herniation and cauda equina syndrome. Considerations on a pathology with different clinical manifestations.

The authors describe six cases of cauda equina syndrome (CES) with different clinical manifestations, etiopathogenetic causes, and degrees of disc prolapse. In three of the cases, the clinical onset was dramatic and acute, in the others it was a hemisyndrome (with acute onset in only 1 case). The site of the disc prolapse was L4-L5 in 3 patients, L3-L4 in 2 patients, and L5-S1 in 1 patient. The authors emphasize the need for the early recognition of the syndrome with a careful clinical examination and history of the patient, as well as timely treatment within 24 hours. For the sake of prevention, and particularly when the hernia is large, the authors suggest monitoring of the stability of the residual disc during surgery and postoperatively the use of a brace for at least one month.

Adult↗

The surgical treatment of hallux valgus using distal metatarsal osteotomy and stabilization with a hallux splint. Preliminary results.

The preliminary results of the surgical treatment of hallux valgus using subcapitate chevron-type osteotomy of the 1st metatarsal modified using a dynamic interfragmentary device are reported. The method satisfies the need to restore angular metatarsophalangeal and intermetatarsal values and improves metatarsal pain caused by hallux valgus correction, which influences the cosmetic results.

Adolescent↗

Manipulative therapy in the treatment of benign cervicobrachialgia of mechanical origin.

The authors compared two methods of rehabilitation, traditional physical therapy and manual therapy, in the treatment of benign cervicobrachialgia of mechanical origin, typical of young subjects, generally consequent to mild and moderate trauma, occasional strain or incorrect posture repeated in time. A MID (minor intervertebral deficit) is at its origin, characterized by the absence of objective instrumental signs (X-ray, CT scan, MRI) and it is only diagnosed based on clinical history and accurate physical examination of the spine segment involved. A sample of 80 patients was divided at random into two groups: the first group was submitted to traditional physiotherapy, the second to manipulative therapy carried out according to the French method of R. Meigne. The results obtained, which were evaluated by univaried ANOVA and Student's "t" test statistical analysis, showed the greater effectiveness of manipulative treatment, in the short term and in the long term.

Adult↗

Reverse HAGL: a possible complication of a tight anterior gleno-humeral stabilization.

Recently the posterior humeral avulsion of the glenohumeral capsule (Reverse HAGL or RHAGL) has been reported in the orthopaedic literature as another possible cause of posterior traumatic shoulder instability. In the present paper we describe three patients in whom a RHAGL was probably a consequence of a tight open anterior shoulder stabilisation. The main complaint of these patients was a stiff shoulder after an open anterior stabilisation. A progressive and worsening discomfort was reported by patients in spite of an accurate rehabilitation program. At persistence of symptoms arthroscopy was performed with evidence of RHAGL that was repaired in association with an anterior release. Clinical and functional improvement was observed but the final outcome seems to be related to range of motion recovery and not only to the posterior repair.

Adult↗