Anidulafungin in Combination with Amphotericin B against Aspergillus fumigatus

We investigated the effects of anidulafungin alone and in combination with amphotericin B against Aspergillus fumigatus. Indifference was the only type of interaction observed in vitro. Anidulafungin at 1 and 5 mg/kg of body weight/day, amphotericin B at 1 mg/kg/day, and combination therapy prolonged the survival of mice with invasive aspergillosis. Anidulafungin at 5 mg/kg/day, alone and in combination with amphotericin B, reduced the kidney fungal burden. Overall, the combination was not superior to the most active single drug.The high mortality rate of invasive aspergillosis has driven recent efforts to determine the efficacy of combination therapy in the treatment and management of those infections (1, 6, 7, 16, 17, 19, 20, 23, 29). Therefore, in this study, the in vitro and in vivo efficacies of the new echinocandin anidulafungin (AFG), alone and in combination with amphotericin B (AMB), against Aspergillus fumigatus were analyzed.Three clinical strains (F2, F3, and F4) isolated from bronchoalveolar lavage specimens from patients with hematological diseases were identified to species level by conventional methods (24).AMB was used as a pure powder (Sigma) for in vitro studies and as a commercial preparation (Fungizone; Bristol-Myers Squibb) for in vivo studies. Pure powder of AFG (Pfizer) was dissolved in dimethyl sulfoxide and further diluted in the test medium or sterile saline solution for in vitro and in vivo studies, respectively.MICs and minimum effective concentrations (MECs; the lowest concentrations that led to the growth of small, rounded, compact hyphal forms compared to the hyphal growth seen in the growth control well) were determined in RPMI 1640 medium by the CLSI M38-A2 broth microdilution method (10, 12).For both susceptibility and checkerboard assays, the MICs and MECs were read visually at 24 and 48 h (10, 25). Drug interactions were classified as synergistic, indifferent, or antagonistic based on the fractional inhibitory concentration (FIC) index (16).Minimum fungicidal concentration (MFC) was considered the concentration of antifungal agents, alone or in combination, that yielded no growth (27).Metabolic activities of conidia and hyphae were assessed in RPMI 1640 medium with l-glutamine, without phenol red and NaHCO₃, by XTT [2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxyanilide inner salt] assay (Tox-2; Sigma) (2, 3, 21).An experimental CD1 mouse (Charles River, Calco, Italy) model of invasive aspergillosis was used by following previously reported procedures (4). A total of three separate in vivo studies were performed by injection of the A. fumigatus F3 isolate. The drug treatments were started 2 h after the infection. AMB at 0.5 and 1 mg/kg of body weight/day, AFG at 1 and 5 mg/kg/day, and combination doses were administered intraperitoneally.In survival studies, the mice were treated daily from day 0 to day 4 and observed for 10 consecutive days.Brain and kidney fungal burdens were determined at day 4 postinfection by CFU count and quantitative PCR based on procedures described by Bowman et al. (5).Histopathology analysis was performed at day 4 postinfection (4). The number of fungal microabscesses was evaluated in 20 consecutive microscopic fields. Each section was classified based on the number of fungal microabscesses as follows: absence, <5, ≥5 to ≤20, and >20.The in vitro results were analyzed by either Mann-Whitney U test or Student''s t test, with a P value of <0.05 considered significant. Survival and tissue burden studies were analyzed by log rank and Mann-Whitney U tests, respectively. Due to multiple comparisons, a P value of <0.016 was considered significant.Our in vitro results are shown in Table ​Table1.1. Overall, AFG MECs were significantly lower than AMB MICs and indifference was the only type of interaction among the two drugs.

TABLE 1.

In vitro susceptibility tests of AFG and AMB, alone and in combination, against three clinical isolates of A. fumigatus^a

Exposure to 3′Sialyllactose-Poor Milk during Lactation Impairs Cognitive Capabilities in Adulthood

Breast milk exerts pivotal regulatory functions early in development whereby it contributes to the maturation of brain and associated cognitive functions. However, the specific components of maternal milk mediating this process have remained elusive. Sialylated human milk oligosaccharides (HMOs) represent likely candidates since they constitute the principal neonatal dietary source of sialic acid, which is crucial for brain development and neuronal patterning. We hypothesize that the selective neonatal lactational deprivation of a specific sialylated HMOs, sialyl(alpha2,3)lactose (3′SL), may impair cognitive capabilities (attention, cognitive flexibility, and memory) in adulthood in a preclinical model. To operationalize this hypothesis, we cross-fostered wild-type (WT) mouse pups to B6.129-St3gal4^tm1.1Jxm/J dams, knock-out (KO) for the gene synthesizing 3′SL, thereby providing milk with approximately 80% 3′SL content reduction. We thus exposed lactating WT pups to a selective reduction of 3′SL and investigated multiple cognitive domains (including memory and attention) in adulthood. Furthermore, to account for the underlying electrophysiological correlates, we investigated hippocampal long-term potentiation (LTP). Neonatal access to 3′SL-poor milk resulted in decreased attention, spatial and working memory, and altered LTP compared to the control group. These results support the hypothesis that early-life dietary sialylated HMOs exert a long-lasting role in the development of cognitive functions.     

Induction of PSA-specific CTLs and anti-tumor immunity by a genetic prostate cancer vaccine

BACKGROUND: Prostate cancer is the most common malignancy in Swedish and American men. Effective curative treatment modalities are debilitating and available only for localized disease. As an immunotherapy approach, DNA encoding prostate-specific antigen (PSA), was used to immunize mice and induce PSA-specific cellular immunity. METHODS: A plasmid expressing PSA, alone or in combination with plasmids coding for GM-CSF and/or IL-2, was used for DNA immunization. Cr-release, intracellular IFN-gamma cytokine staining, and tumor challenge assays were used to evaluate the immune response. RESULTS: The DNA vaccine induces PSA-specific cytotoxic T lymphocytes (CTLs) and when co-injected with IL-2 and GM-CSF it protects four of five mice against a PSA-expressing tumor challenge. CONCLUSIONS: We demonstrate that immunization with a PSA DNA vaccine can evoke PSA-specific cellular immune responses. We also show, for the first time, that a PSA DNA vaccine can induce anti-tumor immunity in vivo.     

Prevalence of abuse of alcohol and other drugs among injured drivers presenting to the emergency department of the University Hospital of Modena, Italy

Among those drivers responsible for injury-producing traffic crashes in a town of northern Italy (Modena) and its surrounding territory, we evaluated the percentage that was positive for alcohol or other drugs affecting CNS function. A total of 115 crash-responsible injured drivers (90 males and 25 females) consecutively presenting to the emergency department at the University Hospital of Modena were enrolled. A urine sample was requested from each driver; the presence of alcohol or drugs was detected by means of various procedures (enzyme immunoassay, liquid or gas chromatography, mass spectrometry). Among the 115 enrolled drivers, 46 (40%) were positive for at least one drug and/or alcohol. Of these 46 drivers, 66% were positive for a single drug, 25% for two drugs, 9% for three or more drugs. Recent use of marijuana was found most frequently (19% out of the total 115 enrolled drivers), surpassing alcohol (10%), amphetamines (7%) and cocaine (6%); 11 drivers (about 10%) tested positive for benzodiazepines. The majority of drivers positive for benzodiazepines were 41-70 years old, while most drivers positive for alcohol or other drugs were 21-40 years old. Thirty-nine (85%) of the positive injured drivers were males and seven (15%) were females. The present data confirm that a significant percentage of injury-producing traffic crashes involves drivers who are under the influence of drugs of abuse, alcohol, or other drugs affecting the CNS.     

Synovial sarcoma enzyme histochemistry of a typical case

Summary A typical case of biphasic synovial sarcoma was studied using enzyme histochemistry. A marked difference between the staining characteristics of the spindle cells and the epithelial-like cells was demonstrated by reactions for various hydrolytic enzymes. The epithelial-like cells exhibited a strong reactivity for alkaline phosphatase, acid phosphatase, adenosine triphosphatase and nonspecific esterase, whereas spindle-cells were completely unreactive when tested for these enzymes.This is, to our knowledge, the first report demonstrating differences in the enzymatic pattern of the two cell populations which compose synovial sarcoma.     

A multifaceted approach to psychodrama in a day treatment center.

Staff at the day treatment center at Norristown (Pa.) State Hospital have found several ways of using psychodrama to facilitate information gathering, diagnostic decision-making, and treatment planning and implementation. They use behavioral techniques such as role rehearsal and modeling. In addition they use techniques such as the mirror image, in vivo dramas, and slight deception to facilitate catharsis. These approaches, which broaden the applicability of psychodrama, are illustrated by case examples.     

Critical Limb Ischemia and Its Impact on Patient Health Preferences and Quality of Life—An International Study

Critical limb ischemia (CLI) has a significant impact on patients'' quality of life (QoL). Despite cost utility evaluations being required by different authorities, data on patient health preferences and utilities for CLI are scarce. Hence, the objective of this study was to assess the impact of CLI on health preferences and health status of affected patients, and to generate health state utilities. In the International Study, 200 patients with CLI (stages III and IV according to Fontaine scale) were interviewed by trained interviewers with a discrete choice instrument, a standard gamble (SG), and the EuroQol-five dimension (EQ-5D) questionnaires (Marten Meesweg, Rotterdam, Netherland). Conjoint analysis showed that a planned amputation (33%) was the most relevant health attribute followed by ambulatory function (25%) and chronic pain (15%). Non-dependence on caregiver impacted on patient health state preference considerably, whereas healing of ulcer/skin lesions had less impact. Preference values obtained from the SG were 0.84, for an amputation subpopulation arriving at 0.70. The EQ-5D index values as well as the EQ-5D visual analog scale for patients with CLI were 0.56. The QoL data of patients with CLI result in decreased QoL and preference values with a planned amputation.     

A case of biliary stones and anastomotic biliary stricture after liver transplant treated with the rendez-vous technique and electrokinetic lithotritor

The paper studies the combined radiologic and endoscopic approach （rendez vous technique） to the treatment of the biliary complications following liver transplant. The ＂rendez-vous＂ technique was used with an electrokinetic lithotripter, in the treatment of a biliary anastomotic stricture with multiple biliary stones in a patient who underwent orthotopic liver transplant. In this patient, endoscopic or percutaneous transhepatic management of the biliary complication failed. The combined approach, percutaneous transhepatic and endoscopic treatment （rendez-vous technique） with the use of an electrokinetic lithotritor, was used to solve the biliary stenosis and to remove the stones. Technical success, defined as disappearance of the biliary stenosis and stone removal, was obtained in just one session, which definitively solved the complications. The combined approach of percutaneous transhepatic and endoscopic （rendez-vous technique） treatment, in association with an electrokinetic lithotritor, is a safe and feasible alternative treatment, especially after the failure of endoscopic and/or percutaneous trans-hepatic isolated procedures.