African histoplasmosis: new clinical and microbiological insights.

African histoplasmosis is defined as the fungal infection caused by Histoplasma capsulatum var. duboisii (Hcd). Studies focused on distinguishing Hcd and H. capsulatum var. capsulatum (Hcc), which coexist in Africa, are scarce or outdated, and African strains are continuously underrepresented. In this work, 13 cases of African patients with histoplasmosis diagnosed in the Spanish Mycology Reference Laboratory have been reviewed showing that 77% had disseminated disease and AIDS as underlying disease although Hcd infection has been classically considered a rare presentation in AIDS patients. Strains isolated from these patients and other clinical and reference strains were studied by assessing classical identification methods and performing a three-loci multi-locus sequence analysis (MLSA). Classical identification methods based on biochemical tests and measurement of yeast size proved to be useless in distinguishing both varieties. The MLSA defined an African cluster, with a strong statistical support, that included all strains with African origin. Finally, mating type was also determined by using molecular methods revealing an unequal mating type distribution in African strains. In conclusion, historical statements and classical identification methods were useless to distinguish between varieties, whereas molecular analyses revealed that all strains with African origin grouped together suggesting that traditional classification should be revised. Further investigation is required in order to unravel traditional concepts about Hcd infection and support results obtained in this work.

Thiamine deficiency, oxidative metabolic pathways and ethanol-induced neurotoxicity: how poor nutrition contributes to the alcoholic syndrome, as Marchiafava-Bignami disease.

Ethanol is an important risk factor for the occurrence of several brain disorders that depend on the amount, period and frequency of its consumption. Chronic use of ethanol often leads to the development of neurodegenerative syndromes, which cause morphological and functional impairments such as foetal alcohol syndrome in newborns exposed to ethanol during pregnancy, Wernicke-Korsakoff Syndrome and, more rarely, Marchiafava-Bignami disease (MBD). MBD is characterized by primary degeneration of the corpus callosum, without inflammation and is associated with oxidative stress and hypovitaminosis, as well as altered mental status, to mention dementia, seizures, depression and so on. This review discusses MBD and poor nutrition as a risk factor for the development of such alcoholic syndrome, with focus on diagnosis, pathogenic aspects, signs and symptoms, as well as therapeutic perspectives. On the basis of the inclusion/exclusion criteria adopted, the performed search in scientific databases (Pubmed, Scielo and Google Scholar) resulted in 100 studies that are being presented and discussed in the present work. Review, case-control and cohort studies on alcoholism-associated hypovitaminosis, oxidative stress, MBD and ethanol metabolism pathways were admitted as relevant. We highlight that MBD is a poorly described, diagnosed, insidious and progressive condition, for which evidence suggests a synergism between ethanol-induced neurotoxic effects and hypovitaminosis B. Present treatment consists of vitamin B1(thiamine) supplementation. Nonetheless, other strategies such as the inclusion of antidepressants or steroidal anti-inflammatories as add-on therapies have been employed as an attempt to improve the damage. Indeed, both the diagnosis and treatment are difficult, and death occurs within few years.

Corrigendum to "First detection of Aspergillus fumigatus azole resistant strain due to Cyp51A TR46/Y121F/T289A in an azole-naive patient in Spain" [New Microbes New Infect 6 (2015) 33-34].

[This corrects the article DOI: 10.1016/j.nmni.2015.04.005.].

Rapid development of Candida krusei echinocandin resistance during caspofungin therapy.

In invasive candidiasis, there has been an epidemiological shift from Candida albicans to non-albicans species infections, including infections with C. glabrata, C. parapsilosis, C. tropicalis, and C. krusei. Although the prevalence of C. krusei remains low among yeast infections, its intrinsic resistance to fluconazole raises epidemiological and therapeutic concerns. Echinocandins have in vitro activity against most Candida spp. and are the first-line agents in the treatment of candidemia. Although resistance to echinocandin drugs is still rare, individual cases of C. krusei resistance have been reported in recent years, especially with strains that have been under selective pressure. A total of 15 C. krusei strains, isolated from the blood, urine, and soft tissue of an acute lymphocytic leukemia patient, were analyzed. Strains developed echinocandin resistance during 10 days of caspofungin therapy. The molecular epidemiology of the isolates was investigated using two different typing methods: PCR-based amplification of the species-specific repetitive polymorphic CKRS-1 sequence and multilocus sequence typing. All isolates were genetically related, and the mechanism involved in decreased echinocandin susceptibility was characterized. Clinical resistance was associated with an increase in echinocandin MICs in vitro and was related to three different mutations in hot spot 1 of the target enzyme Fks1p. Molecular evidence of the rapid acquisition of resistance by different mutations in FKS1 highlights the need to monitor the development of resistance in C. krusei infections treated with echinocandin drugs.

First detection of Aspergillus fumigatus azole-resistant strain due to Cyp51A TR46/Y121F/T289A in an azole-naive patient in Spain.

We report the first isolation of a voriconazole-resistant Aspergillus fumigatus strain harbouring the azole resistance mechanism TR46/Y121F/T289A, recovered from an azole-naive patient in Spain with chronic obstructive pulmonary disease. This new finding in Spain suggests the spread of this resistance mechanism and reinforces the need for antifungal susceptibility surveillance.

Neurological disease in human and canine leishmaniasis--clinical features and immunopathogenesis.

Leishmaniasis is a vectorborne disease caused by Leishmania protozoa, which is a major health problem and a neglected disease common in many regions of the world. Leishmania is an intracellular parasite transmitted by sand flies that causes clinical manifestations ranging from a severe and potentially fatal disease named visceral leishmaniasis to less severe but in many cases disfiguring diseases that mainly affect the skin or mucosal tissues, known as cutaneous leishmaniasis. Despite the detection of Leishmania parasites in the brain and cerebrospinal fluid of human patients and dogs, epidemiological data, as well as information about the mechanisms of central and peripheral nervous system alterations, are poorly described. This review is focused on the current knowledge about the neurological manifestations and immunopathogenic mechanisms in human patients and animals infected with Leishmania.

Desenvolvimento de Periplaneta australasiae Fabricius (Blattodea, Blattidae) em temperatura constante de 30º c e em temperatura ambiente de laboratório / Development of Periplaneta australasiae Fabricius (Blattodea, Blattidae) at a constant temperature of 30° c and at ambient laboratory temperature

Objetivou-se avaliar o desenvolvimento de ovos e ninfas de Periplaneta australasiae (Fabricius) à temperatura de 30 ± 0,2° C, umidade relativa 80 ± 15% e fotofase de 12h e em condições ambientais de laboratório, sem controle de temperatura e umidade relativa; visando à subsídios para medidas de prevenção e controle. As ootecas foram individualizadas em tubos de ensaio até a eclosão. As ninfas foram transferidas para cubas de vidro e alimentadas com ração comercial para coelhos e água ad libitum até a emergência das imagos. Avaliou-se, período de incubação, número de ovos/ ooteca, viabilidade de ovos, número de ninfas/ooteca, período ninfal, viabilidade de ninfas e período ovo/adulto. A diferença do período médio de incubação à temperatura de 30° C (38 dias) e no ambiente (44,5 dias) foi significativa (p < 0,0001); eclodiram, em média 18,1 ninfas/ooteca a 30° C e 21 ninfas/ooteca em condições ambientais (p = 0,006); o período médio de ninfa a 30° C foi de 155,9 dias e no ambiente 279,7 dias (p < 0,0001); a viabilidade de ninfas foi superior a 50%, tanto a 30° C (55,1%) quanto em condições de laboratório (57,2%); no período médio de ovo-adulto de P. australasiae, houve diferença significativa (p< 0,001) entre a temperatura de 30 °C (194,1 dias) e em condições ambientais de laboratório (337,3 dias). Em condições de laboratório, os períodos de incubação, de ninfa e de ovo-adulto de P. australasiae foram aumentados em relação à temperatura de 30° C, não ocorrendo, entretanto, perda nem redução de viabilidade em nenhuma das fases.

With the objective of obtaining standards of measurement for prevention and control, this study compared the development of the eggs and nymphs of Periplaneta australasiae (Fabricius) at a temperature of 30 ± 0.2° C, relative humidity 80 ± 15% and photoperiod of 12 hours versus ambient conditions in the laboratory without controls of temperature and RH. Single ootheca were maintained in test tubes until ecolosion, and nymphs were transferred to glass cubes and fed commercial rabbit ration and water ad libitum until emergence of the imagos. The incubation period, number of eggs/ ootheca, viability of the eggs, number of nymphs/ootheca, nymphal duration, viability of nymphs and duration of egg to adult were all evaluated. The mean difference in the incubation period between the temperature of 30° C (38 days) and ambient conditions (44.5 days) was significant (p < 0.0001); a mean of 18.1 nymphs/ootheca ecloded at 30° C, while 21 nymphs/ootheca ecloded under ambient conditions (p = 0.006); the mean nymphal period at 30° C was 155.9 days while for the ambient it was 279.7 days (p < 0.0001); nymphal viability was greater than 50% for both the 30° C laboratory (55.1%) and the ambient (57.2%); and the mean period from egg to adult of P. australasiae was significantly different (p < 0.001) between the 30° C temperature (194.1 days) and the ambient conditions of the laboratory (337.3 days). Under ambient conditions, the duration of nymphal incubation and egg to adult development of P. australasiae were increased relative to the temperature of 30° C without a reduction in viability in any of the stages.

Mast cell degranulation contributes to susceptibility to Leishmania major.

Leishmaniasis causes high morbidity and mortality in tropical and subtropical areas. Mast cells can be activated by Leishmania or Leishmania products in vitro and in vivo. Several innate immunity mediators, including some released by mast cells, play roles in the outcome of the disease. In this study, we examined whether pharmacological inactivation of mast cells before infection with L. major interferes with the progressive disease in BALB/c mice. The results show that, when mast cells are degranulated before challenge with L. major, susceptible mice become more resistant to infection, as measured by decrease of lesion size and lower parasite loads. Mast cell degranulation reduced IL-4 production. Moreover, mast cells degranulation enhanced mRNA expression for IFN-gamma, inducible nitric oxide, CCL2 and CCL5 in response to infection. Mast cell degranulation also decreased parasite loads in IL-4 KO animals, indicating that mediators other than IL-4 are involved in susceptibility in vivo. Taken together, our results disclose a role for mast cells in the induction of susceptibility to infection. This work contributes to a better understanding of the role of mast cells in Leishmania infection, and suggests a new field of study for strategies to contain the parasite, restricting its dissemination.

The influence of glutathione modulators on the course of Leishmania major infection in susceptible and resistant mice.

Glutathione (GSH) has an important dual role in parasite-host relationship in Leishmania major infection. Our previous studies showed that both antioxidant systems, glutathione and trypanothione/trypanothione reductase, participate in the protection of Leishmania against the toxic effect of nitrogen-derived reactive species. On the other hand, GSH also is very important to the modulation of the effective immune response, inducting NO production and leishmanicidal activity of macrophages. In the present study, we investigated the role of host GSH during the course of L. major infection, analysing the size of footpad lesions and parasite load from mice treated with two GSH modulators, N-acethyl-l-cysteine (NAC) and buthionine sulphoximine (BSO). Resistant mice treated with BSO, which depletes GSH develop exacerbated lesions, but only harbour higher parasite load in their lesions 2 weeks post-infection. Although the NAC treatment does not affect the footpad lesions development in susceptible BALB/c mice, it significantly reduced the tissue parasitism in the lesions throughout the course of infection. Interestingly, the treatment with BSO did not change the course of L. major infection on susceptible mice when compared with nontreated mice. These results suggest that GSH is an important antioxidant modulator during anti-Leishmania immune response in vivo.

[Bacterial cultures and empirical antimicrobial therapy for community-acquired secondary peritonitis]. / Prélèvements microbiologiques et traitements antibiotiques probabilistes des péritonites secondaires communautaires.

Surgeons and anesthetists are frequently confronted with community-acquired secondary peritonitis. We summarize literature results and consensus conferences concerning the types of bacteriologic sampling and cultures and the empiric choice of an antibiotic regimen based on the probable pathogens encountered in community-acquired secondary peritonitis. These studies leave some doubt as to the necessity for routine blood cultures and the need for anaerobic cultures of peritoneal fluid. No one disputes the need for broad spectrum antibiotic therapy, but there is no consensus regarding one, two, or three drug antibiotic regimens or whether an aminoglycoside is an essential part of the recipe. Duration of antibiotic therapy is still a subject of controversy with recommendations varying from 24 hours to 10 days. The need for antibiotics with activity against enterococcus and the need for systematic antifungal therapy when fungal growth is noted in the peritoneal fluid remain undefined. These uncertainties underline the need for treating physicians within each establishment to elaborate a written consensus of antibiotic therapy.

Microcystin-LR promote intestinal secretion of water and electrolytes in rats.

We showed previously that exposure to microcystin-LR causes renal toxic effects in isolated perfused rat kidney, and that inflammatory mediators from supernatants of macrophages stimulated by microcystin-LR are involved in this process. The aim of this research was to examine water and electrolytes secretion in vivo, induced by microcystin-LR and supernatant of macrophages stimulated for this toxin (SUP.MphiS + MCLR), using perfused rat ileal segment and ligated intestinal loop models. We found microcystin-LR at 1 microg/ml (0.09 +/- 0.003* vs. control 0.07 +/- 0.001 g of secretion/2 cm of loop; P < 0.05*) and the SUP.MphiS + MCLR after 18 h postinoculation (0.10 +/- 0.003 vs. control 0.03 +/- 0.002 g/cm) caused intestinal secretion. In addition, microcystin-LR caused significant sodium secretion (-2.18 +/- 0.72* vs. control 2.18 +/- 0.50 microEq g(-1) min(-1)), potassium (-0.26 +/- 0.04* vs. control 0.32 +/- 0.03 microEq g(-1) min(-1)), chloride (MCLR = -3.29 +/- 1.93* vs. control 0.88 +/- 1.25 microEq g(-1) min(-1)) and water (-0.012 +/- 0.004* vs. control 0.002 +/- 0.002 ml g(-1) min(-1)). We also demonstrated SUP.MphiS + MCLR to induce intestinal secretion of electrolytes (sodium, potassium, chloride) and water. These findings suggested that microcystin-LR and lamina propria macrophages-derived mediators are able to induce intestinal secretion in vivo, probably via inhibition of protein phosphatase.

Sinus tissue pharmacokinetics after oral administration of amoxicillin/clavulanic acid.

OBJECTIVES: The in vitro synergy of the amoxicillin/clavulanic acid combination has not always translated in vivo into clinical superiority compared with amoxicillin alone. Specifically, conflicting reports have disputed the superiority of the combination in the treatment of both acute otitis media and acute sinusitis. One possible reason for this may have to do with inadequate target tissue pharmacokinetics. To explore this possibility in the sinuses, we undertook the present investigation. STUDY DESIGN: A randomized, open, single-dose, sinus tissue pharmacokinetic study with oral amoxicillin/clavulanic acid. METHODS: Twenty-three adult patients with chronic rhinosinusitis who had been selected for surgery were randomly allocated to receive a tablet of 875/125 mg amoxicillin/clavulanate 2 to 4 hours before surgery began. During the operation tissue samples were collected at specific sinonasal sites for determination of both amoxicillin and clavulanic acid concentration levels. RESULTS: Amoxicillin displayed adequate tissue levels throughout the sinuses, high enough to cover common susceptible pathogens. However, the presence of clavulanate was detected in only half of the sinonasal tissue samples. CONCLUSIONS: The kinetics of oral clavulanic acid apparently fails to provide a widespread anti-beta-lactamase activity capable of enhancing the activity of amoxicillin in all parts of the sinuses. Despite this, amoxicillin/clavulanic acid maintains a central role in the treatment of acute rhinosinusitis, because amoxicillin is still the most effective oral beta-lactam against Streptococcus pneumoniae, a particularly virulent and increasingly resistant upper respiratory tract pathogen. Also, as our data show, a concomitant anti-beta-lactamase activity can be expected to occur, although in an unpredictable fashion.

Penetration of cefuroxime into chronically inflamed sinus mucosa.

OBJECTIVES: Despite its seeming relevance, limited information exists about antibiotic sinus tissue penetration and how it is affected by inflammation. Thus the reason for the present investigation. STUDY DESIGN: A randomized, open, multiple-dose, pharmacological study, employing cefuroxime axetil, an approved oral antimicrobial for the treatment of acute bacterial rhinosinusitis, was developed. METHODS: Twenty subjects, selected for surgery because of chronic rhinosinusitis, were randomly allocated to receive either a short (3-8 d) or a long (9-14 d) preoperative treatment regime with 500 mg cefuroxime axetil BID, the last dosage being taken 3 to 4 hours before surgery. At the operation, tissue samples were collected at specific sinonasal sites for both pharmacological determination of antibiotic levels and histopathological assessment of the degree of inflammation. The blood levels of the drug were simultaneously assayed. RESULTS: Cefuroxime kinetic behavior on chronically inflamed mucosa was shown to be, for the most part, dependent on the blood levels, regardless of the inflammatory state. Distribution was even throughout the different sinus cavities, and the tissue levels were still, 3 to 4 hours after dosing, above the reported minimum inhibitory concentration (MIC) values for some of the most prevalent sinus pathogens. The extended treatment course did not seem to add any extra histopathological or pharmacological benefit. CONCLUSIONS: Cefuroxime penetrates adequately and uniformly into chronically inflamed sinus mucosa, apparently unaffected by the degree of inflammation, in a way not dissimilar to its pharmacokinetic behavior in the normal state. Persistent MIC levels for common pathogens still warrant antimicrobial efficacy for a significant period of time after dosing.

Laccase from Trametes versicolor: stability at temperature and alkaline conditions and its effect on biobleaching of hardwood kraft pulp.

The enzyme laccase was produced by the white-rot fungus Trametes versicolor in repeated batches cultures with immobilized mycelium. Two different culture conditions were used. Enzymes produced were evaluated regarding their stability at high temperatures (55 degrees C and 65 degrees C) and at alkaline conditions (pH 7.0 and pH 8.0) having in view the application of these enzymes in biobleaching of hardwood Kraft pulp. Biobleaching experiments were divided in two parts, enzymatic prebleaching followed by chemical bleaching. In the enzymatic prebleaching the enzyme laccase was used at two conditions of pH and temperature, whereas the reaction time was fixed at 1 h in all pretreatments. In the chemical bleaching the DEDED and DEpDED sequences were used. The enzyme action was evaluated by Kappa number, viscosity, and brightness at the end of bleaching sequences. There were obtained values of Kappa numbers lower than control assays, viscosities compatible with industrial pulps, and brightness higher than controls, when pulps were pretreated for 1 h with laccase at pH 8.0 and 55 degrees C.

Production and characterization of Phanerochaete chrysosporium lignin peroxidases for pulp bleaching.

The production of lignin peroxidase from Phanerochaete chrysosporium was studied using immobilized mycelia in nylon-web cubes in semicontinuous fermentation using glucose pulses or ammonium tartrate pulses. Consistent enzyme production was achieved when glucose pulses were used, leading to an average activity of 253 U/L. The crude enzyme was added to eucalyptus kraft pulp before conventional and ECF bleaching sequences. Optimization of the enzymatic pretreatment led to the following operational conditions: enzyme load of 2 U/g of pulp, hydrogen peroxide addition rate of 10 ppm/h, and reaction time of 60 min. Pulp final characteristics were dependent on the chemical treatment sequence that followed enzymatic pretreatment. The chief advantage of enzymatic pretreatment was pulp viscosity preservation, which was observed in most of the experiments carried out with seven different chemical treatment sequences.

Pulp bleaching using laccase from Trametes versicolor under high temperature and alkaline conditions.

Kraft pulp was delignified using laccase produced by the white rot fungus Trametes versicolor immobilized in solid support under specific conditions. The stability tests showed that this enzyme was stable for 6 h at 55 degrees C and pH 8.0, allowing its use under pH and temperature conditions very close to those used in industrial bleaching. In this work, unbleached hardwood Kraft pulp was submitted to prebleaching using 2 U laccase/g pulp basis. Reaction time, temperature, and pH of the enzymatic treatment were investigated. Good results regarding Kappa number reduction, selectivities, and high viscosities were obtained when prebleaching was performed for 1 h at temperature of 55 degrees C and pH 8.0 followed by alkaline extraction and ECF bleaching sequences.

Recorrência de adenocarcinoma em coto gástrico: qual o papel da cirurgia? / Recurrence of remnant gastric adenocarcinoma: what is the importance of surgery?

Os autores apresentam sua experiência com recorrência de adenocarcinoma em coto gástrico, perfazendo um total de cinco casos no período de 1981 a 1989, dos quais somente um caso foi passível de cirurgia com ressecçäo de intençäo curativa, com sobrevida de quatro meses. Fica-nos a impressäo de que o screening pós-operatório pode levar a aumento de taxas de re-ressecabilidade e sobrevida em casos precoces

Original Article: Changes in Platelet Function, von Willebrand Factor and Factor VIII in Patients Undergoing Aorto-bifemoral By-pass with Dacron Grafts.

Platelets play a major role in the development of patency complications in vascular grafts. The aim of this study was to evaluate changes in platelet count and function, and also in factor VIII:C (FVIII:C) and von Willebrand factor (vWF) plasma levels, induced by aorto-bifemoral by-pass with Dacron grafts in seven patients. Platelet count, platelet aggregate ratio (PAR), and platelet aggregability induced by several stimuli, as well as FVIII:C and vWF plasma levels were evaluated before and on days 1,4,9 and 11 after surgery. We observed a mild thrombocytopenia on day 1, followed by a progressive increase in platelet count, which attained a relative thrombocytosis on the 11th day. PAR did not vary significantly during the whole observation period. Platelet aggregation, assayed by the optical method using ADP, epinephrine, arachidonic acid, collagen and ristocetin, (decreased on days 1 and 4). Thereafter, an increase in aggregation was observed until day 11 when hyperaggregability was verified. FVIII:C and vWF peaked on the 4th day, decreasing progressively to pre-surgery values on day 11.