Molecular characterisation and antifungal susceptibility of clinical Cryptococcus deuterogattii (AFLP6/VGII) isolates from Southern Brazil.

Cryptococcosis, caused by Cryptococcus gattii sensu lato, is an emerging disease that was initially found in (sub)tropical regions but recently expanded to temperate regions. Cryptococcus gattii s.l. infections are mostly encountered in healthy individuals, frequently affecting both lungs and the central nervous system (CNS). Usually, C. gattii s.l. is less susceptible to antifungal compounds than its counterpart, C. neoformans s.l. We studied 18 clinical C. gattii s.l. isolates with amplified fragment length polymorphism (AFLP) fingerprinting, mating-typing, multi-locus sequence typing (MLST) and antifungal susceptibility testing. All isolates were C. deuterogattii (genotype AFLP6/VGII), 14 were mating-type α and four were type a. Amphotericin B, itraconazole, voriconazole, posaconazole and isavuconazole showed high activity, with minimum inhibitory concentration (MIC) ranges of 0.063-0.25, 0.031-0.25, 0.031-0.25, 0.031-0.25 and <0.016-0.25 µg mL-1, respectively. Fluconazole and flucytosine had high geometric mean MICs of 2.07 and 3.7 µg mL-1, respectively. Most cases occurred in immunocompetent patients (n = 10; 55.6 %) and CNS involvement was the most common clinical presentation (n = 14; 77.8 %). Three patients (16.7 %) showed sequelae, hyperreflexia, dysarthria, diadochokinesia, anosmia and upper limb weakness. In conclusion, all infections were caused by C. deuterogattii (AFLP6/VGII) and the majority of patients were immunocompetent, with the CNS as the most affected site. All antifungal drugs had high in vitro activity against C. deuterogattii isolates, except fluconazole and flucytosine.

Persistent altered spermatogenesis in long-term childhood cancer survivors.

This study evaluated male gonadal function in long-term survivors of childhood cancer and assessed the suitability of offering sperm analysis to all those patients independently of the diagnosis and treatment received. A total of 43 survivors of acute lymphoblastic leukemia (21), acute myeloid leukemia (1), neuroblastoma (8), ganglioneuroblastoma (1), ganglioneuroma (2), Wilms' tumor (9), and mesoblastic nephroma (1) underwent sperm analysis at a mean age of 20.2 years, after a mean time off treatment of 13.6 years. Eight of the patients (19%) were azoospermic, 2 (5%) were severely oligo-asthenozoospermic, and only 16 (37%) were normozoospermic. A control group of healthy volunteers aged < or = 30 years included no azoospermic subjects, 7% severely oligo-asthenozoospermic, and 67% normozoospermic. Comparisons were also made with patients treated at our Human Reproductive Unit aged < or = 30 years (n = 373) whose percentages for the above parameters were 4, 9, and 42%, respectively. Cumulated cyclophosphamide dose and basal follicle-stimulating hormone (FSH) levels were identified as independent factors associated with azoospermia or severe oligo-asthenozoospermia. Azoospermic and severely oligo-asthenozoospermic survivors had significantly smaller mean testicular volume and higher basal FSH levels than the other survivors, but small testicles (sum of both testicular volume < or = 20 mL) and/or abnormally high basal FSH (> 10 mIU/mL) were present in only half of the azoospermic survivors. Male long-term survivors of childhood cancer constitute a high-risk subpopulation for altered sperm analysis. It seems justified to offer sperm analysis to all long-term survivors.

Cluster versus conventional immunotherapy in patients allergic to Dermatophagoides pteronyssinus: a controlled study of in vivo and in vitro parameters.

We designed a cluster schedule of immunotherapy for patients allergic to Dermatophagoides pteronyssinus which showed good safety and clinical efficacy. Here we compare the in vivo and in vitro changes with those of a conventional schedule in a controlled trial. Sixty-three patients were randomized as follows: 29 were treated with the cluster schedule, 15 with a conventional schedule and 19 received no immunotherapy. A standardized extract was used. Changes in in vivo parameters (skin prick test and conjunctival provocation test) and in in vitro parameters (IgE, IgG, IgG1 and IgG4 for the complete extract, Der p 1 and Der p 2) were measured before immunotherapy (T0), on reaching maintenance phase (T1), and after 6 (T2), 12 (T3) and 18 months of maintenance (T4). Cutaneous reactivity showed a significant decrease from T1 in both the cluster and conventional schedules, and conjunctival reactivity was also significantly lowered from T1 in these groups. Specific IgE decreased and specific IgG, IgG1 and IgG4 increased significantly from T1 in the cluster and conventional schedules. Neither of these parameters showed any changes in the group without immunotherapy. In conclusion, our cluster schedule induced changes in cutaneous and conjunctival reactivity and in immunological parameters that were similar to those achieved with the conventional schedule; these changes did not appear in patients who did not undergo immunotherapy.

Dermatophagoides pteronyssinus cluster immunotherapy. A controlled trial of safety and clinical efficacy.

We designed a cluster schedule of immunotherapy for patients allergic to Dermatophagoides pteronyssinus to reach the maximal recommended dose in 7 weeks. We compared its safety and clinical efficacy with those of a conventional schedule in a controlled trial. Sixty-three patients were randomized as follows: 29 were treated with the cluster schedule, 15 with a conventional schedule and 19 without immunotherapy. A standardized extract was used. Clinical efficacy was measured by visual analog scale, clinical severity score, symptom/medication diary cards and control of peak expiratory flow (PEF) in asthmatic patients, before immunotherapy (T0), on reaching the maintenance phase (T1), and after 6 (T2), 12 (T3) and 18 months of maintenance (T4). The safety of immunotherapy was found to be good. Visual analog scale improved significantly at T4 in the cluster and conventional schedules, and clinical severity score also improved from T1 in these schedules. Diary cards improved from T1 in the cluster schedule and from T2 in the conventional schedule in asthmatic patients. Significant improvements in diary cards in rhinitis patients and PEF were found only in the cluster schedule. There were no changes in the group without immunotherapy. In conclusion, our cluster schedule showed as good tolerance and clinical efficacy as the conventional schedule in patients allergic to D. pteronyssinus. These clinical improvements did not appear in the group without immunotherapy.

[Solid ovarian tumors in childhood]. / Tumores sólidos de ovario en la infancia.

We evaluate our experience in solid ovarian tumors at the Children's Hospital La Fe. The main aims of the study were finding out relevant clinical data for a correct clinical approach, and the optimal surgical attitude in order to obtain tumor relieving with preservation of fertility when possible. A review of 28 girls under 14 years with solid ovarian tumors was carried out. Previous history, initial symptoms and signs, analytical and radiological explorations, treatment and evolution were analysed. 17 benign and 11 malignant tumors were found. Main clinical data were acute abdominal pain and palpation of a mass. Surgery was performed in all of them: salpyngo-oophorectomy in 23 and oophorectomy in 5, with tumoral resection. All patients survive, with periodic follow-up. We remark the frequency of acute abdominal pain as initial symptom of an ovarian tumor, and the importance of a surgical attitude that, intending to be curative, would preserve fertility.

Renaturation of blotted allergens increases the sensitivity of specific IgE detection.

Several authors have demonstrated that renaturation is an essential step for the appropriate recognition of blotted proteins. The use of nonionic detergents has been described as a useful alternative to enhance the antigenicity in immunoblotting, although elution from proteins by detergents has been observed. To measure the influence of different factors on the sensitivity of specific IgE by immunoblotting, we used twenty human sera from atopic patients who were allergic or nonallergic to a common, reliable allergen (grass pollen mixture). The use of Nonidet-P40 was found to be a useful alternative for the renaturation of the allergens. No elution from the membrane was found when employing this detergent, even at high concentrations (3%), and its use gave better sensitivity than methanol. On the other hand, we detected that methanol possessed renaturing properties. A transfer method using diffusion instead of electric transfer gave the best results and two membranes could be obtained from each gel. Using this method, we found that after NP-40 incubation of the membrane, the use of bovine albumin could be omitted as blocking agent and that its use had even deleterious effects.

Neuroblastoma IV-S followed by extra-adrenal pheochromocytoma 15 years later.

Common origin of sympathoblasts and pheochromoblasts from the neural crest cells is generally accepted. Neuroblastoma and pheochromocytoma result from an abnormal proliferation of these committed cells. They are included in the group of neuroendocrine neoplasms, formerly named Apudomas. Previous reports of mixed tumours of neuroblastoma and pheochromocytoma, and ganglioneuroma and pheochromocytoma, support this hypothesis. A case of extra-adrenal pheochromocytoma in an adolescent who 15 years before was successfully treated for a stage IV-S neuroblastoma without evidence of primitive tumour is reported. Two hypothesis are formulated: (1) the second neoplasm may have arisen from the unlocated primitive tumour, probably silent evolution could be due to its maturation into ganglioneuroma-pheochromocytoma; (2) taken into account a common origin of both neoplasms, the patient may have a genetic predisposition to develop neuroendocrine tumours. Whatever the mechanism, second neoplasms, as the case reported, late relapses, and late toxic effects may justify long follow-up of neuroblastoma survivors.

Airborne contact dermatitis from Frullania.

7 subjects, each giving a history of scaly or vesicular dermatitis in light-exposed areas after going for strolls to the park, the forest or the country during warm weather, were evaluated. Patch testing was carried out with Frullania dilatata and Frullania tamarisci as well as with 2 common members of the Compositae and to alantolactone and the sesquiterpene lactone mix. All the patients were positive to F. dilatata, 3 reacted to F. tamarisci as well, and 2 to florists' chrysanthemum. All of them except 1 gave positive responses to the sesquiterpene lactone mix, but only 3 reacted to alantolactone. Airborne contact dermatitis from these species of Frullania was considered to be the cause of their eczema. Patch testing with the sesquiterpene lactone mix seems to be a good screen for Frullania sensitivity; however, a variable pattern of response to these sesquiterpene-lactone-containing materials was observed, suggesting individual immunologic responses to them.

A prospective safety-monitoring study of immunotherapy with biologically standardized extracts.

We evaluated the safety of immunotherapy in 419 patients who attended our allergy department for treatment. They were suffering from rhinitis and asthma caused by sensitization to grass pollen or Dermatophagoides pteronyssinus. Immunotherapy was given by biologically standardized aluminum hydroxide adsorbed extracts according to a conventional schedule. Local reactions were recorded in 10.5% of the patients and systemic reactions in 4.8%. Only 0.37% of the doses administered were associated with systemic side-effects. We found that 84% of the patients who showed systemic reactions were asthmatic subjects (P < 0.01), and most of them were sensitized to D. pteronyssinus (71%). Side-effects occurred more frequently during the dose-increase period (P < 0.05). After 9482 doses had been administered, no anaphylactic shock or life-threatening reactions were registered. We believe the risk associated with immunotherapy to be drastically reduced when treatment is carefully monitored by skilled personnel. In such conditions, as shown by our study, immunotherapy is safe.

Occupational asthma and contact urticaria from dried flowers of Limonium tataricum.

Dried flowers of Limonium tataricum (LT) are widely used for semipermanent floral arrangements. We report a case of a floral industry worker in whom exposure to LT precipitated rhinoconjunctivitis, asthma, and contact urticaria. Studies revealed the presence of immediate skin test reactivity and positive leukocyte histamine release in response to LT extract. Specific anti-LT antibodies were detected in the patient's serum by direct RAST. Bronchial challenge with LT extract also resulted in an isolated immediate asthmatic response. Both the positive methacholine inhalation test and the significant changes observed in PEFR measurements when the patient handled LT supported the diagnosis of occupational asthma. These findings strongly suggest that an IgE-mediated immunologic mechanism is responsible for the patient's respiratory and cutaneous symptoms.

Fine-needle aspiration biopsy in children: experience in 70 cases.

Results of 70 fine-needle aspiration biopsies (FNAB) were evaluated retrospectively in 61 pediatric patients. Over a period of 9 months all mass lesions suspected being malignant were aspirated. Twelve of the 70 aspirations were performed in children having known tumours, in order to exclude recurrence or metastasis. The others were carried out to obtain a diagnosis. Satisfactory specimens were obtained from 58 (83%). There were 21 benign diagnoses, 36 malignant diagnoses, and 1 with suspected malignancy. Correlation of histologic and cytologic diagnoses was possible in 45 cases. The diagnostic sensitivity and specificity were 95% and 80%, respectively. We have found FNAB more accurate in the diagnosis of malignancies than in benign lesions. The results suggest that this is a useful technique for obtaining a first diagnosis of malignancy, as well as for excluding recurrence or metastatic disease.