In vitro effects of naloxone on T-lymphocyte-dependent antibacterial activity in hepatitis C virus (HCV) infected patients and in inflammatory bowel disease (IBD) patients

Naloxone acts as an opioid antagonist, displacing opioid drugs from cellular receptors. Among opioid substances, β-endorphins are able to bind to several cell receptors, even including those expressed by immune cells. In this respect, evidence has been provided that in the course of viral infections, as well as in patients with ulcerative colitis high levels ofβ-endorphins are detectable. Here, peripheral blood lymphocytes (PBL) from 21 HCV infected patients and 14 patients with IBD, respectively, were incubated with Naloxone and Naloxone + Ca²⁺ in order to evaluate a putative modulation of PBL-mediated antibacterial activity. In fact, previous studies have demonstrated a reduction of this T-cell activity in HCV and IBD patients.

In general terms, the above treatment led to a recovery of the depressed antibacterial activity. In some cases, increase in T lymphocyte function was obtained with Naloxone alone, while in other cases the combination Naloxone + Ca²⁺ gave rise to a restorative effect. Of note, in some instances, lymphocytes were unresponsive to pharmacological modulation.

The overall results suggest that β-endorphins may down modulate T-cell antibacterial response in HCV and in IBD patients by saturating peripheral receptors on immune cells. Therefore, it is likely that Naloxone and/or Naloxone + Ca²⁺ may displace opioid drugs, thus antagonizing their effects.     

Effects of follicular fluid supplementation of in-vitro maturation medium on the fertilization and development of equine oocytes after in- vitro fertilization or intracytoplasmic sperm injection

The aim of this study was to compare the effect of the addition of follicular fluid (FF) collected from preovulatory follicles with that of oestrous mare serum (EMS) (acting as the control) to TCM-199 medium on the in-vitro maturation, fertilization and development of equine cumulus-enclosed oocytes. Oocytes (<30 mm in diameter) were obtained from the ovaries of slaughtered mares. After in-vitro maturation in the presence of the two supplements, their fertilization, cleavage and developmental potential were compared after conventional in-vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) using frozen-thawed spermatozoa. Follicular fluid did not increase the maturation of oocytes to metaphase II stage compared to control. After IVF, there was no difference in fertilization rates between FF- supplemented oocytes and controls (7/87, 8.4% of oocytes showing two pronuclei with FF versus 7/116, 6% with EMS; not significant). However, after ICSI, FF-supplemented oocytes showed significantly increased normal fertilization (32/85, 37.6% of two-pronuclear oocytes) and developmental potential (15/31, 48% cleavage) compared to the control oocytes (7/47, 14.9%, P < 0.01; and 2/48, 4%, P < 0.01, respectively). Overall, ICSI resulted in increased fertilization rates compared to IVF, regardless of the presence or absence of FF (39/132, 29.5% with ICSI versus 14/203, 6.9%). These results suggest that follicular fluid supplementation may improve the maturity of equine cumulus-enclosed oocytes sufficiently for the successful use of ICSI, but not sufficiently for normal sperm-egg interaction occurring during IVF.      

The spontaneous bacterial peritonitis in cirrhotic patients. To a new gold standard.

Of 282 consecutive ascites prospectively collected in 54 months, Spontaneous Bacterial Peritonitis (SBP) was diagnosed in 8.5% of the cases, "probable" SBP in 31.1%, Bacteriascites (BA) in 3.5% and Sterile Ascites (SA) (negative ascitic fluid culture with PMN less than 250/mm3) in 74.8%. Escherichia Coli (41.6%) and Staphylococcus Epidermidis (60%) were the most frequent pathogens isolated in patients with SBP and BA, respectively. With regards to in-hospital mortality, 18% of patients with BA and 50% with SBP died; the mortality seemed to be related to the degree of hepatic and renal damage, to a higher peripheral and ascitic WBC concentration and to a lower pH of ascitic fluid (FA). When the comparative analysis was applied to the four groups of ascites, a different distribution of clinical signs and biohumoral parameters appeared. As a matter of fact, abdominal pain, fever and rebound tenderness resulted significantly more frequent in SBP and "probable" SBP. Furthermore, the mean values of peripheral and ascitic WBC concentration, of serum creatinine and of ALT were statistically higher in SBP and "probable" SBP than in SA and BA groups. The strict relationship, both symptomatologic and biochemical, between SA and BA on the one hand and between "probable" SBP and SBP on the other, prompted us to conclude that "probable" SBP and SBP represent different patterns of the same disease. Therefore, the subclassification in the four groups outlined above would not be in accordance with the clinical practice and could give rise to the physician's confusion and uncertainty.     

Role of beta-endorphin on phospholipase production in Malassezia pachydermatis in dogs: new insights into the pathogenesis of this yeast.

Malassezia spp. are lipophilic yeasts that are part of the normal cutaneous microflora and sometimes act as pathogens causing dermatitis. This study investigated the interactions occurring between beta-endorphin and phospholipase activity in isolates of M. pachydermatis in dogs presenting cutaneous lesions. Phospholipase production was evaluated and quantified on 144 isolates suspended in Dixon broth to which different beta-endorphin concentrations (from 600 to 0.6 pM) were added. The isolates were divided into three groups: group A comprised isolates from lesional skin of dogs with dermatitis confined to one site, group B consisted of isolates from the healthy skin of the same dogs with localized lesions, and group C was made up of isolates from assorted skin sites of healthy dogs. A statistically higher phospholipase activity than that of the controls was recorded in group B at all tested beta-endorphin concentrations. In groups A (Pz=0.62) and C (Pz=0.62) phospholipase activity was statistically higher than the controls only at a concentration of 600 pM. This study suggests that beta-endorphin plays an important role in the production of phospholipase in M. pachydermatis isolates and provides evidence that beta-endorphin concentrations affect the number but not the Pz value of phospholipase-producing isolates. B-endorphin concentrations may play a relevant role in inducing M. pachydermatis cell differentiation towards the production or non-production of phospholipase.     

First demonstration of an increased serum level of reactive oxygen species during the peripartal period in the ewes

Reactive Oxygen Species (ROS) are produced during oxidative metabolism, and regulate many biological processes. The acute inflammation characterizing parturition induces many physiological changes. Among them, there is evidence that ROS affect the synthesis of many factors involved in parturition. Our study aims to determine serum levels of ROS in periparturient ewes, as well as to establish a value of reference of their physiological concentration. ROS determination was performed on blood collected every 12 hours in periparturient twin pregnant ewes. Our results will show a significant increase in ROS concentrations from the beginning to the end of the experiment. This increase may be due to the inflammatory process establishing during parturition.     

Biological monitoring of hospital personnel occupationally exposed to antineoplastic agents

To detect trace amounts of urinary cyclophosphamide (CP), ifosfamide (IF) and methotrexate (MTX), sensitive and specific high-performance liquid chromatography/ tandem mass spectrometry (HPLC-MS/MS) procedures, incorporating either liquid-liquid (for CP and IF), or solid-phase, extraction (for MTX) have been developed. Urinary platinum (Pt) was also detected using inductively coupled plasma-mass spectrometry (ICP-MS). These methods showed acceptable imprecision and inaccuracy. The limit of detection (LOD) was 50 ng/l for CP and IF, 200 ng/l for MTX and 1 ng/l for Pt. Biomonitoring was performed on two consecutive days on nine subjects preparing, and seven administering, antineoplastic drugs. Urine was collected at the beginning, at the end and during the work shift. Eighteen urine samples were positive for CP (range: 50-10031 ng/l), whereas IF was detected in one subject only (153 ng/l). LOD was never exceeded for MTX. In urine samples from nurses and pharmacy technicians, Pt was detected in three subjects (range 920-1300 ng/l). These findings were compared with the results from a previous survey carried out in the same hospital when different work practices were in use. The proposed methods are simple, fast and reliable and can be used to identify exposure of hospital personnel handling antineoplastic drugs.     

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.     

Reversal of cachexia in patients treated with potent antiretroviral therapy

The introduction of HAART has changed the nutritional status of HIV patients. In the pre-protease inhibitor (PI) era, more than 60% of HIV-positive persons presented with protein energy malnutrition (PEM) and vitamin and mineral deficit. This caused progressive physical-metabolic wasting (wasting syndrome/cachexia) and increased susceptibility to opportunistic infections and drug toxicity. PEM was a concurrent cause in 80% of deaths attributed to AIDS. Since 1996, the year in which PIs were introduced, the number of patients dying as a result of AIDS has decreased by two thirds, and cachexia is no longer the AIDS terminal phase in developed countries. But different patterns of nutritional status changes have appeared in association with the use of newer anti-HIV therapies and with longer survival of HIV-infected patients. A new clinical and laboratory syndrome--lipodystrophy syndrome--now affects patients receiving PI-based therapy. This syndrome consists of changes in body shape that are caused by an abnormal redistribution of fat. Fat accumulates in the abdominal area (truncal and visceral obesity), in the axillary pads (bilateral symmetric lipomatosis), and in the dorsocervical pads ("buffalo hump," "bull neck") but decreases in the legs, arms, and nasolabial and cheek pads (peripheral lipodystrophy). Hyperlipidemia and insulin resistance are also frequently present (metabolic syndrome X). Pathogenic mechanisms of lipid and fat tissue disturbances are discussed in this article, and the clinical approach to patient management and therapeutic options for lipodystrophy and lipid dysmetabolism is evaluated.