TiO2-MWCNT nanohybrid: Cytotoxicity, protein corona formation and cellular internalisation in RTG-2 fish cell line.

Titanium dioxide nanoparticles-multiwalled carbon nanotubes (TiO2-MWCNT) nanohydrid has an enhanced photocatalytic activity across the visible light with promising applications in environmental remediation, solar energy devices and antimicrobial technologies. However, it is necessary to evaluate the toxicological effects of TiO2-MWCNT towards safe and sustainable development of nanohybrids. In this work, we studied the cytotoxicity, protein corona formation and cellular internalisation of TiO2-MWCNT on fibroblasts derived from gonadal rainbow trout tissue (RTG-2) for the first time. This nanohydrid did not show any toxicity effect on RTG-2 cells up to 100 mg L-1 after 24 h of exposure as monitored by alamar blue, neutral red and trypan blue assays (in presence or absence of foetal bovine serum, FBS). Futhermore, cryo-transmission electron microscopy analysis demonstrated that TiO2 particles is attached on nanotube surface after FBS-protein corona formation in cell culture medium. Raman spectroscopy imaging showed that TiO2-MWCNT can be internalised by RTG-2 cells. This work is a novel contribution towards better understanding the nanobiointeractions of nanohydrids linked to their in vitro effects on fish cells in aquatic nanoecotoxicology.

Primary Ciliary Dyskinesia in a Portuguese Bronchiectasis Outpatient Clinic.

Primary ciliary dyskinesia (PCD) is a rare hereditary condition characterized by decreased mucociliary clearance of the airways and a compromised reproductive system, resulting in male and female infertility. Several mutations with varied clinical and pathological features have been documented, making diagnosis a challenging process. The purpose of this study is to describe the clinical and pathological features of Portuguese patients with PCD and to examine their genetic variants. A retrospective observational analysis was conducted with patients who were being monitored at a bronchiectasis outpatient clinic in 2022 and had a confirmed or high-likelihood diagnosis of PCD. In total, 17 patients were included in the study, with 12 (66.7%) having PCD confirmed and 5 (29.4%) having a high-likelihood diagnosis. Furthermore, 12 patients were subjected to transmission electron microscopy (TEM), with 7 (58.3%) exhibiting one hallmark defect. Genetic test data was obtained for all 17 patients, with 7 of them (41.2%) displaying a pathogenic/likely pathogenic mutation in homozygosity. To summarize, PCD is an uncommon but significant hereditary illness with consequences regarding morbidity and mortality. Despite the lack of a specific treatment, it is critical to confirm the diagnosis with genetic testing in order to effectively manage the disease and its accompanying disorders.

X-ray spectrometry imaging and chemical speciation assisting to understand the toxic effects of copper oxide nanoparticles on zebrafish (Danio rerio).

Currently, copper nanoparticles are used in various sectors of industry, agriculture, and medicine. To understand the effects induced by these nanoparticles, it is necessary to assess the environmental risk and safely expand their use. In this study, we evaluated the toxicity of copper oxide (nCuO) nanoparticles in Danio rerio adults, their distribution/concentration, and chemical form after exposure. This last assessment had never been performed on copper-exposed zebrafish. Such evaluation was done through the characterization of nCuO, acute exposure tests and analysis of distribution and concentration by microstructure X-ray fluorescence spectroscopy (µ-XRF) and atomic absorption spectroscopy (GF-AAS). Synchrotron X-ray absorption spectroscopy (XAS) was performed to find out the chemical form of copper in hotspots. The results show that the toxicity values of fish exposed to nCuO were 2.4 mg L-1 (25 nm), 12.36 mg L-1 (40 nm), 149.03 mg L-1 (80 nm) and 0.62 mg L-1 (CuSO4, used as a positive control). The total copper found in the fish was in the order of mg kg-1 and it was not directly proportional to the exposure concentration; most of the copper was concentrated in the gastric system. However, despite the existence of copper hotspots, chemical transformation of CuO into other compounds was not detected.

Investigation of acute toxicity, accumulation, and depuration of ZnO nanoparticles in Daphnia magna.

Size is a key factor controlling the rate of dissolution of nanoparticles, such property can be explored for producing controlled release fertilizers. Hence, one can expect the increasing discharge of nanoparticles closer to water streams in the near future. In this study, we employed the model fresh water organism Daphnia magna to investigate the uptake, acute toxicity and depuration of ZnO nanoparticles. The present study shows that the median lethal concentration (LC50) depended on particle size and the presence of surfactant. The LC50 for positive control ZnSO4 (2.15 mg L-1), 20 nm ZnO (1.68 mg L-1), and 40 nm ZnO (1.71 mg L-1) were statistically the same. However, the addition of surfactant increased the LC50 of 40 nm and 60 nm to 2.93 and 3.24 mg L-1, respectively. The 300 nm ZnO was the least toxic nanoparticle presenting LC50 of 6.35 mg L-1. X-ray fluorescence chemical imaging revealed that Zn accumulated along the digestive system regardless the particle size. Finally, contrary to what have been reported by several papers, the present study did not detect any depuration of ZnO nanoparticles in the next 24 h past the exposure assays. Thus, the ability of organisms to expel ingested nanomaterials might be dependent on specific physical-chemical features of such nanomaterials.

Transition from APAP to CPAP may be a cost-effective health intervention in OSA patients.

OBJECTIVE: Obstructive sleep apnea (OSA) is a common disorder associated with a significant economic burden. Continuous positive airway pressure (CPAP) and auto-titrating positive airway pressure (APAP) are recognized therapeutic options in patients with OSA, although treatment costs are higher with APAP. We conducted a study aimed at evaluating the effectiveness and potential cost savings resulting from the implementation of a protocol guiding the transition to CPAP in OSA patients previously treated with APAP. METHODS: This prospective study included patients with OSA under APAP who were followed up at the Sleep Medicine outpatient clinic of a tertiary referral hospital between January 2019 and January 2021. Treatment was switched to CPAP in patients who met the following criteria: satisfactory adaptation and adherence to APAP, residual apnea-hypopnea index (AHI) of < 5/hour, and no relevant air leaks. APAP and CPAP outcomes were compared and an estimate of the savings obtained by the transition from APAP to CPAP was calculated. RESULTS: Ninety-three patients were included in the study. APAP and CPAP were both effective in correcting obstructive events and improving daytime sleepiness. No significant differences were found regarding treatment adherence and tolerance between both PAP modalities. The selection of fixed-pressure CPAP through 90th or 95th percentile APAP pressure proved to be effective and an alternative strategy to titration polysomnography. At the end of this two-year study, the transition from APAP to CPAP enabled savings of at least 10,353€. CONCLUSION: The transition from APAP to CPAP may be an effective, well-tolerated, safe, and cost-saving strategy in patients with OSA.

Role of the antioxidant defense system during the production of lignocellulolytic enzymes by fungi

Fungi are used for the production of several compounds and the efficiency of biotechnological processes is directly related to the metabolic activity of these microorganisms. The reactions catalyzed by lignocellulolytic enzymes are oxidative and generate reactive oxygen species (ROS). Excess of ROS can cause serious damages to cells, including cell death. Thus, the objective of this work was to evaluate the lignocellulolytic enzymes produced by Pleurotus sajor-caju CCB020, Phanerochaete chrysosporium ATCC 28326, Trichoderma reesei RUT-C30, and Aspergillus niger IZ-9 grown in sugarcane bagasse and two yeast extract (YE) concentrations and characterize the antioxidant defense system of fungal cells by the activities of superoxide dismutase (SOD) and catalase (CAT). Pleurotus sajor-caju exhibited the highest activities of laccase and peroxidase in sugarcane bagasse with 2.6 g of YE and an increased activity of manganese peroxidase in sugarcane bagasse with 1.3 g of YE was observed. However, P. chrysosporium showed the highest activities of exoglucanase and endoglucanase in sugarcane bagasse with 1.3 g of YE. Lipid peroxidation and variations in SOD and CAT activities were observed during the production of lignocellulolytic enzymes and depending on the YE concentrations. The antioxidant defense system was induced in response to the oxidative stress caused by imbalances between the production and the detoxification of ROS

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Role of the antioxidant defense system during the production of lignocellulolytic enzymes by fungi.

Fungi are used for the production of several compounds and the efficiency of biotechnological processes is directly related to the metabolic activity of these microorganisms. The reactions catalyzed by lignocellulolytic enzymes are oxidative and generate reactive oxygen species (ROS). Excess of ROS can cause serious damages to cells, including cell death. Thus, the objective of this work was to evaluate the lignocellulolytic enzymes produced by Pleurotus sajor-caju CCB020, Phanerochaete chrysosporium ATCC 28326, Trichoderma reesei RUT-C30, and Aspergillus niger IZ-9 grown in sugarcane bagasse and two yeast extract (YE) concentrations and characterize the antioxidant defense system of fungal cells by the activities of superoxide dismutase (SOD) and catalase (CAT). Pleurotus sajor-caju exhibited the highest activities of laccase and peroxidase in sugarcane bagasse with 2.6 g of YE and an increased activity of manganese peroxidase in sugarcane bagasse with 1.3 g of YE was observed. However, P. chrysosporium showed the highest activities of exoglucanase and endoglucanase in sugarcane bagasse with 1.3 g of YE. Lipid peroxidation and variations in SOD and CAT activities were observed during the production of lignocellulolytic enzymes and depending on the YE concentrations. The antioxidant defense system was induced in response to the oxidative stress caused by imbalances between the production and the detoxification of ROS.

Swyer-James-Macleod syndrome as a rare cause of unilateral hyperlucent lung: Three case reports.

RATIONALE: Swyer-James-Macleod syndrome (SJMS) is a rare lung disorder characterized by unilateral hyperlucent lungs which arises as a complication of bronchiolitis obliterans. It is typically diagnosed during childhood, although some patients may only be diagnosed in adulthood, often as an incidental finding, but others due to recurrent chest infections. PATIENT CONCERNS: Three patients were referred to our institution with complaints of dyspnea on exertion, chronic productive cough, and recurrent pulmonary infections. Two of them had a history of lower respiratory tract infections during childhood. DIAGNOSIS: A computed tomography scan was performed and showed unilateral reduced density of the lung and bronchiectasis in the 3 patients. Based on the clinical presentation and radiologic features, the diagnosis of SJMS was established. INTERVENTIONS: Patients started inhaled corticosteroids and long acting beta agonist, vaccines to prevent respiratory infections were administered and airway clearance techniques were taught. OUTCOMES: Two patients became asymptomatic with inhaled therapy and no recurrent chest infections were observed over a 3-year follow-up period, being discharged from our institution to the general practitioner. The 3rd patient had some improvement in the frequency of pulmonary infections and dyspnea, without improvement in respiratory function tests, maintaining vigilance at our center. LESSONS: These cases highlight the importance of being aware of this condition and its frequent association with bronchiectasis, which may adversely affect the prognosis, to manage patients appropriately and prevent recurrent pulmonary infections.

Effects of sewage sludge application on unfertile tropical soils evaluated by multiple approaches: A field experiment in a commercial Eucalyptus plantation.

Sewage sludge (SS) reuse in forest plantation as soil fertilizer/amendment has tremendously increased in recent years. However, SS may have high concentrations of potentially toxic elements (PTE), representing a potential risk for soil and the whole ecosystem. This paper was aimed to assess the toxicity of PTE in unfertile tropical soils amended with SS in a commercial Eucalyptus plantation, with an integrated multiple approaches combining: i) the use of a battery of bioassays (Daphnia magna, Pseudokcrichirella subcapitata, Lactuca sativa, and Allium cepa); and ii) the evaluation of some PTE (Cd, Cr, Cu, Fe, Mn, Ni, Pb, and Zn) and their availability into the pedoenvironment. Differences in total and available PTE between SS doses and time of treatments were evaluated using ANOVA; correlations between PTE and bioassays by a sparse partial robust M-regression (SPRM), while multiple correlations among parameters were performed by principal factor analysis (PFA). Results show that PTE contents in soils tended to increase with SS application doses. However this cannot be assumed as a general rule since in all the investigated treatments the PTE concentrations were consistently below both soil natural background concentrations and quality reference values. Bioassays showed a generalized low eco- and genotoxicity of SS with an increase in toxicity at increasing SS doses but with a clear decreasing trend as time went by. A. cepa was the most sensitive bioassay followed by P. subcapitata > D. magna > L. sativa. Overall, the results indicate that in realistic open field conditions SS risk may be lower than expected due to dynamic decrease in PTE toxicity with time after application. This study has an important implication that open-field trials should be strongly encouraged for evaluating environmental risk of SS application in forestry.

Toxicity assessment of TiO2-MWCNT nanohybrid material with enhanced photocatalytic activity on Danio rerio (Zebrafish) embryos.

The increasing production and use of nanomaterials is causing serious concerns about their safety to human and environmental health. However, the applications of titanium dioxide nanoparticles (TiO2NP) and multiwalled carbon nanotubes (MWCNT) hybrids has grown considerably, due to their enhanced photocatalytic efficiency. To our knowledge, there are no reports available to the scientific community about their toxicity. In this work, we perform a toxicity assessment of TiO2NP and TiO2-MWCNT nanohybrid materials using Zebrafish embryos standardized 96 h early life stage assay, under different exposure conditions (with and without UV light exposure). After exposure the parameters assessed were acute toxicity, hatching rate, growth, yolk sac size, and sarcomere length. In addition, µ-probe X-ray fluorescence spectroscopy (µ-XRF) was employed to observe if nanoparticles were uptaken by zebrafish embryos and consequently accumulated in their organisms. Neither TiO2NP nor TiO2-MWCNT nanohybrids presented acute toxicity to the zebrafish embryos. Moreover, TiO2NP presents sublethal effects for total length (with and without UV light exposure) on the embryos. This work contributes to the understanding of the potential adverse effects of the emerging nanohybrid materials towards safe innovation approaches in nanotechnology.

Amazoninema gen. nov., (Synechococcales, Pseudanabaenaceae) a novel cyanobacteria genus from Brazilian Amazonian rivers.

The genus Leptolyngbya includes morphotypes with thin cells and simple morphology, and is one of the most common cyanobacterial genera found in a wide range of environments. In many cases, however, the morphotypes assigned to this genus do not share a common ancestor based on 16S rRNA gene phylogeny, which has led to the description of novel genera, such as Nodosilinea, Oculatella, Pantanalinema, Alkalinema, Thermoleptolyngbya, Onodrimia, Timaviella and Toxifilum. Thus, four novel isolates, with a comparable morphology to Leptolyngbya, were recovered from the Amazon and Solimões rivers. The novel 16S rRNA gene sequences obtained from these strains were placed together as a new and distinct phylogenetic lineage that is more closely related to the clusters embracing the genera Nodosilinea, Haloleptolyngbya and Halomicronema than to the genus Leptolyngbya. Additionally, these novel 16S rRNA gene sequences showed similarity values lower than 95 % compared with those from the most phylogenetic related groups and/or established genera. Altogether, these results supported the erection of a novel genus, named Amazoninema, to accommodate the novel isolates. Likewise, a comparison of their 16S rRNA gene sequences revealed similarities higher than 99.8 %, indicating that they belong to a single species, which was corroborated by analysing their 16S-23S internal transcribed spacer regions and unique Box-B helix pattern. Few studies have been undertaken to uncover the cultured diversity of cyanobacteria from Amazonia, and to our knowledge, this is the first cyanobacteria genus erected, considering morphotypes isolated exclusively from Brazilian Amazonian rivers.

Degradation of textile dyes by cyanobacteria.

Dyes are recalcitrant compounds that resist conventional biological treatments. The degradation of three textile dyes (Indigo, RBBR and Sulphur Black), and the dye-containing liquid effluent and solid waste from the Municipal Treatment Station, Americana, São Paulo, Brazil, by the cyanobacteria Anabaena flos-aquae UTCC64, Phormidium autumnale UTEX1580 and Synechococcus sp. PCC7942 was evaluated. The dye degradation efficiency of the cyanobacteria was compared with anaerobic and anaerobic-aerobic systems in terms of discolouration and toxicity evaluations. The discoloration was evaluated by absorption spectroscopy. Toxicity was measured using the organisms Hydra attenuata, the alga Selenastrum capricornutum and lettuce seeds. The three cyanobacteria showed the potential to remediate textile effluent by removing the colour and reducing the toxicity. However, the growth of cyanobacteria on sludge was slow and discoloration was not efficient. The cyanobacteria P. autumnale UTEX1580 was the only strain that completely degraded the indigo dye. An evaluation of the mutagenicity potential was performed by use of the micronucleus assay using Allium sp. No mutagenicity was observed after the treatment. Two metabolites were produced during the degradation, anthranilic acid and isatin, but toxicity did not increase after the treatment. The cyanobacteria showed the ability to degrade the dyes present in a textile effluent; therefore, they can be used in a tertiary treatment of effluents with recalcitrant compounds.

Degradation of textile dyes by cyanobacteria

Abstract Dyes are recalcitrant compounds that resist conventional biological treatments. The degradation of three textile dyes (Indigo, RBBR and Sulphur Black), and the dye-containing liquid effluent and solid waste from the Municipal Treatment Station, Americana, São Paulo, Brazil, by the cyanobacteria Anabaena flos-aquae UTCC64, Phormidium autumnale UTEX1580 and Synechococcus sp. PCC7942 was evaluated. The dye degradation efficiency of the cyanobacteria was compared with anaerobic and anaerobic-aerobic systems in terms of discolouration and toxicity evaluations. The discoloration was evaluated by absorption spectroscopy. Toxicity was measured using the organisms Hydra attenuata, the alga Selenastrum capricornutum and lettuce seeds. The three cyanobacteria showed the potential to remediate textile effluent by removing the colour and reducing the toxicity. However, the growth of cyanobacteria on sludge was slow and discoloration was not efficient. The cyanobacteria P. autumnale UTEX1580 was the only strain that completely degraded the indigo dye. An evaluation of the mutagenicity potential was performed by use of the micronucleus assay using Allium sp. No mutagenicity was observed after the treatment. Two metabolites were produced during the degradation, anthranilic acid and isatin, but toxicity did not increase after the treatment. The cyanobacteria showed the ability to degrade the dyes present in a textile effluent; therefore, they can be used in a tertiary treatment of effluents with recalcitrant compounds.

The aquatic impact of ionic liquids on freshwater organisms.

Ionic liquids (ILs), also known as liquid electrolytes, are powerful solvents with a wide variety of academic and industrial applications. Bioassays with aquatic organisms constitute an effective tool for the evaluation of ILs' toxicity, as well as for the prediction and identification of possible moieties that act as toxicophores. In this work, the acute toxicity of six ILs and two commonly used organic solvents was evaluated using freshwater organisms: Daphnia magna, Raphidocelis subcapitata and Hydra attenuata. The bioassays were performed by exposing the organisms to increasing concentrations of the ILs and observing D. magna immobilization, R. subcapitata growth inhibition, and the morphological or mortality effects in H. attenuata. The results demonstrate that the tested organisms are not equally susceptible to the ILs, e.g., bmpyr [BF4] was the least toxic compound for R. subcapitata, N1,1 [N1,1,1OOH] for D. magna and emim [Tf2N] for H. attenuata. This highlights the importance of applying a battery of assays in toxicological analysis. Additionally, Hydra proved to be the most tolerant species to the tested ILs. According to their hazard rankings, the tested ILs are considered practically harmless or moderately toxic, except (Hex)3(TDec)P [Cl], which was classified as highly toxic. The ILs were revealed to be more harmful to aquatic systems than the tested organic solvents, reaffirming the need to analyze carefully the (eco)toxicological impact of these compounds. The present study provides additional data in the evaluation of the potential hazard and the impact of ILs in the environment.

Optimization of lipids production by Cryptococcus laurentii 11 using cheese whey with molasses.

This study aimed the optimization of culture condition and composition for production of Cryptococcus laurentii 11 biomass and lipids in cheese whey medium supplemented with sugarcane molasses. The optimization of pH, fermentation time, and molasses concentration according to a full factorial statistical experimental design was followed by a Plackett-Burman experimental design, which was used to determine whether the supplementation of the culture medium by yeast extract and inorganic salts could provide a further enhancement of lipids production. The following conditions and composition of the culture medium were found to optimize biomass and lipids production: 360 h fermentation, 6.5 pH and supplementation of (g L(-1)): 50 molasses, 0.5 yeast extract, 4 KH2PO4, 1 Na2HPO4, 0.75 MgSO4 · 7H2O and 0.002 ZnSO4 · H2O. Additional supplementation with inorganic salts and yeast extract was essential to optimize the production, in terms of product concentration and productivity, of neutral lipids by C. laurentii 11. Under this optimized condition, the production of total lipids increased by 133% in relation to control experiment (from 1.27 to 2.96 g L(-1)). The total lipids indicated a predominant (86%) presence of neutral lipids with high content of 16- and 18-carbon-chain saturated and monosaturated fatty acids. This class of lipids is considered especially suitable for the production of biodiesel.

Optimization of lipids production by Cryptococcus laurentii 11 using cheese whey with molasses

This study aimed the optimization of culture condition and composition for production of Cryptococcus laurentii 11 biomass and lipids in cheese whey medium supplemented with sugarcane molasses. The optimization of pH, fermentation time, and molasses concentration according to a full factorial statistical experimental design was followed by a Plackett-Burman experimental design, which was used to determine whether the supplementation of the culture medium by yeast extract and inorganic salts could provide a further enhancement of lipids production. The following conditions and composition of the culture medium were found to optimize biomass and lipids production: 360 h fermentation, 6.5 pH and supplementation of (g L-1): 50 molasses, 0.5 yeast extract, 4 KH2PO4, 1 Na2HPO4, 0.75 MgSO4•7H2O and 0.002 ZnSO4•H2O. Additional supplementation with inorganic salts and yeast extract was essential to optimize the production, in terms of product concentration and productivity, of neutral lipids by C. laurentii 11. Under this optimized condition, the production of total lipids increased by 133% in relation to control experiment (from 1.27 to 2.96 g L-1). The total lipids indicated a predominant (86%) presence of neutral lipids with high content of 16- and 18- carbon-chain saturated and monosaturated fatty acids. This class of lipids is considered especially suitable for the production of biodiesel.

Comparison of two lipid extraction methods produced by yeast in cheese whey

This work aimed to evaluate nine strains of yeast, previously identified as good producers of lipids in honey medium, for selecting the most suitable strain for the production of lipids in cheese whey medium and compared two well known extraction methods of lipids from the culture medium. The highest yield of total lipids was 1.27 g.L-1 produced by Cryptococcus laurentii 11. A comparison was made between the two culture media: cheese whey and liquid YEPG, and two lipid extraction methods: Bligh and Dyer and Folch et al. for C. laurentii. The experiments were performed with 2² full factorial design using two factors and two levels. Lipid content was higher in cheese whey and there was no difference in the extraction methods statistically. The method of Bligh and Dyer was used in preference to Folch et al. as it resulted in larger mean of total lipids.

Genetics correlates with lung function and nocturnal ventilation in myotonic dystrophy.

BACKGROUND: Dystrophia myotonica (DM) is the most frequent adult-onset muscular dystrophy. Type 1 is caused by the cytosine-thymine-guanine (CTG) repeat expansion in the DM protein kinase gene. Respiratory muscle weakness and altered central ventilatory control lead to hypercapnia and lung volume restriction. PURPOSE: This study aims to review the respiratory involvement in DM patients and study its relation with genetics. METHODS: Retrospective study of patients with DM referred for respiratory assessment was made. Noninvasive ventilation (NIV) was considered to daytime hypercapnia or symptoms of nocturnal hypoventilation. RESULTS: Forty-two consecutive patients (37.9 ± 13.6 years) were evaluated. Mean CTG length was 642.8 ± 439.2 repeats. In the first evaluation, mean forced vital capacity (FVC) was 74.4 ± 20.2 %, maximal expiratory pressure (MEP) 35 ± 16 %, maximal inspiratory pressure 52 ± 23 %, peak cough flow (PCF) 327.3 ± 97.7 L/min, arterial pressure of oxygen 79.7 ± 11.3 mmHg, arterial pressure of carbon dioxide 45.5 ± 6.2 mmHg, overnight minimal peripheral oxygen saturation (SpO2) 79.6 ± 11.6 %, and apnea-hypopnea index 13.9 ± 9.9. CTG length was found to be related with MEP (r = -0.67; p = 0.001) and SpO2 (r = -0.37; p = 0.039). NIV was started in 25 patients. Ventilated patients had lower FVC (2.19 to 3.21 L; p < 0.001) and PCF (285.3 to 388.5 L/min; p = 0.003) and more CTG repeats (826.6 to 388.5 repeats; p = 0.02). NIV compliance was poor in seven patients (28 %) and related with hypercapnia (r = 0.87; p = 0.002) and inspiratory positive airway pressure setting (r = 0.65; p = 0.009). Ventilation improved symptoms and nocturnal hypoventilation. Comparing the first and last evaluations, only PCF was significantly lower (275.0 to 310.8 L/min; p = 0.019). CONCLUSIONS: Ventilatory insufficiency is very common in patients with DM and CTG length may be useful to predict it. Prolonged NIV improves symptoms, nocturnal hypoventilation and maintains daily blood gases. Routine evaluation of PCF should not be forgotten and assisted coughing training provided.

Cerebral tuberculomas - A clinical challenge.

Cerebral tuberculomas are a rare and serious form of tuberculosis (TB) due to the haematogenous spread of Mycobacterium Tuberculosis (MT). Symptoms and radiologic features are nonspecific, leading sometimes to misdiagnosis. Anti-TB drugs are essential for the successful treatment of cerebral tuberculomas but there is no agreement regarding the duration of therapy. The authors present a case of a 55 years old male, presented to the emergency room with sudden onset of diplopia. Cerebral computerized tomography revealed multiple brain lesions, with contrast enhancement and peri-lesional oedema. The patient was HIV negative and because of previous malignancy the first suspicion was metastatic disease. Cultural exam of the bronchial wash showed MT sensitive to all first-line drugs. The patient started antituberculosis treatment with 4 drugs (HRZE) for 2 months, followed by maintenance therapy (HR). Treatment was prolonged for 24 months because at 12th and 18th months of treatment one of the brain lesions, although significantly smaller, still showed contrast enhancement. Even though it is not clear if contrast enhancement lesions represent active lesions or just inflammation, continuing treatment until total resolution of the tuberculomas is probably prudent.