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

Postural asymmetry and lateralized rotation in normal rats administered apomorphine

The study examines the hypothesis that the direction of circling exhibited by normal rats administered apomorphine reflects a drug-induced bias to use one specific hindleg for stepping and the other one for postural support. Results indicate that a majority of rats injected with this dopamine receptor agonist do show an asymmetry in the usage of the hindlegs (postural bias). However, the side of more frequent steppings does not predict the direction of circling. Moreover, postural bias may exist without lateralized circling. It is suggested that lateralized circling and postural asymmetry are two of several biases induced by apomorphine that may coexist in individual rats.     

Behavioral consequences of bone marrow transplantation in the treatment of murine mucopolysaccharidosis type VII.

The gusmps/gusmps mouse is a model of the human lysosomal storage disease mucopolysaccharidosis type VII caused by deficient beta-glucuronidase activity. Bone marrow transplantation has been shown to correct some of their biochemical and pathological abnormalities but its efficacy in correcting their neurological functional deficits is unknown. We transplanted the neonatal gusmps/gusmps mice and their normal controls and evaluated their central nervous system function with two behavioral tests: the grooming test, a developmentally regulated and genetically based activity, and a Morris water maze test which assessed spatial learning abilities. The two transplanted groups groomed less than the normals, were unable to remember the location of an invisible platform from day to day, and were severely impaired at developing strategies to locate the platform in unfamiliar locations. The performance of both normal and mutant transplanted groups was clearly inferior to the untreated normals and, in some instances, close to or worse than the untreated mutants, even though the enzyme abnormalities of the mutants have been partially corrected. Hence, the behavioral deficits in the mutant mice were not restored to normal while similarly treated normal mice showed significant functional deterioration, indicating the detrimental consequence of this therapy in the neonatal period.     

On the causal association between exposure to leather dust and nasal cancer: further evidence from a case-control study.

A case-control study was performed on the incident cases of nasal cavity tumours which occurred between 1968 and 1982 among the residents of Vigevano (Lombardy region, northern Italy). This area is characterised by a high prevalence of shoemakers (especially in leather); the activity has predominated in Vigevano since the beginning of this century. Twenty one cases were identified (16 men and five women); 20 were histologically confirmed as nasal epithelial tumours; 17 had already died at the time of interview and the occupational history was obtained from the next of kin. Two controls per case were selected from the general population and matched by vital status, age, sex, and residence. The overall odds ratio for the subjects exposed to leather dust was 47.1 for men and 3.5 for women. The odds ratio was higher for adenocarcinoma and among the workers exposed to the worst working conditions. A significant trend for the level of exposure to leather dust was found. Nevertheless, even the jobs characterised by a relatively low exposure were found to have a significantly higher risk (OR = 7.5). Smoking habits and exposure to solvents are unlikely to confound the relation between exposure to leather and nasal tumours.     

Evidence against a role of the rat's dorsal noradrenergic bundle in selective attention and place memory

In Experiment 1,6-hydroxydopamine-induced lesions of the dorsal noradrenergic bundle (DNB) in rats did not impair either acquisition of non-delayed alternation, retention of non-delayed alternation, or performance of alternation with delays in a T-maze, whether or not the goal arms of the maze were visually distinctive. These results were in contrast with those of a previous report indicating that DNB lesions impair learning of spatial alternation. In Experiment 2, the lack of a reliable effect of DNB lesions on learning performance of spatial alternation was confirmed. However, the rats with DNB lesions showed an impairment of spontaneous alternation. The negative results of the present study do not support either the hypothesis that depletion of forebrain noradrenaline impairs selective attention or the hypothesis that such depletion induces an amnesia for past places. On the other hand, the finding of impaired spontaneous alternation is consistent with previous observations suggesting that depletion of forebrain noradrenaline impairs habituation of fear reactions.     

Phenoxy herbicides and soft-tissue sarcomas in female rice weeders. A population-based case-referent study

A population-based case-referent study was conducted in an area of northern Italy where rice growing is the predominant agricultural activity and phenoxy herbicides have been used since 1950. Manual rice weeding was formerly performed by a seasonal female working population; in the early 1950s these women were concurrently exposed to chemical herbicides. Sixty-eight persons representing incident and histologically revised cases (31 women) and 158 population referents (73 women) were interviewed. The cases were histologically confirmed independently by two blinded pathologists, and exposure to phenoxy herbicides was assessed by two blinded pesticide researchers. An age-adjusted odds ratio of 0.91 was found for the living men (with suspect exposures; no man diagnosed as a case had been exposed with certainty to phenoxy herbicides). Among the living women the relative risk was 2.7 (90% confidence interval 0.59-12.37), and it further increased when attention was restricted to women exposed in the whole 1950-1955 period and to younger age groups.     

Impact of a soy drink on climacteric symptoms: an open-label,crossover, randomized clinical trial

Objectives: The objective of this study is to evaluate the effects of a soy drink with a high concentration of isoflavones (ViveSoy®) on climacteric symptoms.

Methods: An open-label, controlled, crossover clinical trial was conducted in 147 peri- and postmenopausal women. Eligible women were recruited from 13 Spanish health centers and randomly assigned to one of the two sequence groups (control or ViveSoy®, 500?mL per day, 15?g of protein and 50?mg of isoflavones). Each intervention phase lasted for 12 weeks with a 6-week washout period. Changes on the Menopause Rating Scale and quality of life questionnaires, as well as lipid profile, cardiovascular risk and carbohydrate and bone metabolism were assessed. Statistical analysis was performed using a mixed-effects model.

Results: A sample of 147 female volunteers was recruited of which 90 were evaluable. In both sequence groups, adherence to the intervention was high. Regular consumption of ViveSoy® reduced climacteric symptoms by 20.4% (p?=?0.001) and symptoms in the urogenital domain by 21.3% (p?p?<0.05).

Conclusion: Regular consumption of ViveSoy® improves both the somatic and urogenital domain symptoms of menopause, as well as health-related quality of life in peri- and postmenopausal women.     

Colorectal cancer risk and nitrate exposure through drinking water and diet

Ingested nitrate leads to the endogenous synthesis of N‐nitroso compounds (NOCs), animal carcinogens with limited human evidence. We aimed to evaluate the risk of colorectal cancer (CRC) associated with nitrate exposure in drinking water and diet. A case‐control study in Spain and Italy during 2008‐2013 was conducted. Hospital‐based incident cases and population‐based (Spain) or hospital‐based (Italy) controls were interviewed on residential history, water consumption since age 18, and dietary information. Long‐term waterborne ingested nitrate was derived from routine monitoring records, linked to subjects’ residential histories and water consumption habits. Dietary nitrate intake was estimated from food frequency questionnaires and published food composition databases. Odd ratios (OR) were calculated using mixed models with area as random effect, adjusted for CRC risk factors and other covariables. Generalized additive models (GAMs) were used to analyze exposure‐response relationships. Interaction with endogenous nitrosation factors and other covariables was also evaluated. In total 1,869 cases and 3,530 controls were analyzed. Average waterborne ingested nitrate ranged from 3.4 to 19.7 mg/day, among areas. OR (95% CIs) of CRC was 1.49 (1.24, 1.78) for >10 versus ≤5 mg/day, overall. Associations were larger among men versus women, and among subjects with high red meat intake. GAMs showed increasing exposure‐response relationship among men. Animal‐derived dietary nitrate was associated with rectal, but not with colon cancer risk. In conclusion, a positive association between CRC risk and waterborne ingested nitrate is suggested, mainly among subgroups with other risk factors. Heterogeneous effects of nitrate from different sources (water, animal and vegetables) warrant further research.