Multifaceted intervention of Bacillus spp. against salinity stress and Fusarium wilt in tomato.

AIM: This study aimed to screen halotolerant Bacillus strains able to promote growth and protect tomato plants against salt stress and Fusarium wilt (Fusarium oxysporum f. sp. lycopersici). METHODS AND RESULTS: We evaluated some halotolerant strains of Bacillus spp. (Bacillus velezensis (AP-3) and Bacillus spp. (AP-6, AP-85 and AP-100)) to promote growth of tomato plants grown under salinity stress conditions and to protect them against Fusarium wilt disease. Such strains had been previously selected among 154 bacterial strains through biochemical tests (siderophores and indoleacetic acid productions, cellulase and catalase activity, nitrogen fixation and phosphate solubilization) in the presence of 100-mmol l-1 NaCl. Besides the above-mentioned strains, B. subtilis QST-713 (SerenadeTM ) was also evaluated. Compared to control plants, aboveground dry weight increased in plants inoculated with AP-6, AP-85, AP-3, AP-100 and QST-713 strains developed in the absence of salt stress. The same tendency occurred for root dry weight; however, AP-3 strain was more effective, promoting an increase of 163%, when compared to control. Chlorophyll index and height increased >40 and 53%, respectively, for all Bacillus strains. Saline stress reduced plant growth regardless of the presence of Bacillus. Height, stem diameter, and aboveground and root dry weights increased in plants treated with Bacillus strains grown under saline conditions when compared to control. Bacillus velezensis AP-3 reduced the severity of Fusarium wilt in tomato by 50% when compared to control. CONCLUSION: Halotolerant Bacillus strains controlled tomato Fusarium wilt, increased growth as well as tolerance to salt stress. SIGNIFICANCE AND IMPACT OF THE STUDY: We demonstrated the efficacy of halotolerant Bacillus strains to control Fusarium wilt and improve tomato growth. We also demonstrated that these Bacillus strains protect tomato plants against salt stress. Bacillus can be used in an eco-friendly way because they are considered Generally Recognized As Safe.

Potentiation of antifungal activity of terbinafine by dihydrojasmone and terpinolene against dermatophytes.

Dermatophytoses are infections that affect keratinized tissues. Their main etiologic agents are fungi of the genera Microsporum and Trichophyton. The emergence of resistant fungi and the clinical relevance of dermatophytosis have encouraged studies that aim to increase the arsenal of drugs or act on mechanisms that confer multiple drug resistance. This study investigated the modulating activity of terbinafine promoted by dihydrojasmone and terpinolene against Microsporum canis LM 216, Trichophyton interdigitale H6 and T. interdigitale Δmdr2. The minimum inhibitory concentration (MIC) of test drugs was determined by broth microdilution. The effect of the drugs tested on plasma membrane functionality was analysed. Terbinafine MIC was determined in sub-inhibitory concentrations of monoterpenes. Finally, it was performed an association study with terbinafine and monoterpenes. Dihydrojasmone presented lower MIC values than terpinolene. All fungi were sensitive to terbinafine, starting at 1 µg ml-1 . All tested drugs increased K+ release (P < 0·05), affecting the functionality of the plasma membrane. Dihydrojasmone modulated the sensitivity of all strains against terbinafine, and terpinolene modulated the sensitivity of M. canis LM 216 and T. interdigitale Δmdr2. The monoterpenes and terbinafine drug associations presented synergism. In conclusion, the results suggest that the dihydrojasmone and terpinolene are promising antifungal agents that potentiate the antifungal activity of terbinafine against dermatophytes.

Obstructive sleep apnea and biomarkers of inflammation in ischemic stroke.

BACKGROUND AND OBJECTIVES: Despite a confirmed association between obstructive sleep apnea (OSA) and stroke, the pathogenesis of OSA in stroke has not been hitherto clarified. The aim of this study was to evaluate the relationship between respiratory abnormalities and atherogenic pro-inflammatory markers, interleukin-1beta (IL-1ß), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α) in acute ischemic stroke patients. METHODS: Nocturnal polygraphy was performed in 50 consecutive patients with acute ischemic stroke in the first week after the event. Levels of inflammatory markers (IL-6, IL-1ß and TNF-α) were determined from morning blood samples and comparatively analyzed between cases with and without severe OSA and with age-matched controls. RESULTS: All patients with acute ischemic stroke, 31 men, mean age (64.3 ± 7.7 years), had apnea-hypopnea index (AHI) > 5 and 35 (70%) had severe OSA (AHI ≥ 30). Hypertension was more frequent in patients with severe OSA (85.7%) when compared to controls (40.0%) (P = 0.002). Stroke outcome, assessed by the Barthel index, tended to be more severe (P = 0.06) in cases with severe OSA. Patients with mild/moderate OSA and with severe OSA showed higher levels of IL-6 when compared to controls (P = 0.01 and P = 0.000, respectively). Among cases with acute stroke and severe OSA, IL-6 levels were correlated with lower oxyhemoglobin desaturation (r=-0.30; P = 0.001) and with the desaturation index (r = 0.15; P = 0.02). CONCLUSIONS: IL-6, an atherogenic marker, shows a commensurate increase in stroke patients with OSA. It is correlated with oxyhemoglobin desaturation and with desaturation index and may be a surrogate measure to evaluate continuous positive airway pressure therapy.

Role of the haem oxygenase/carbon monoxide pathway in Clostridium difficile toxin A-induced enteritis in mice.

Clostridium difficile is the major cause of antibiotic-associated colitis, a disease with significant morbidity and mortality. This study investigated the role of the haem oxygenase-1 (HO-1)/carbon monoxide (CO) pathway in C. difficile toxin A-induced enteritis in mice. The HO substrate haemin, zinc protoporphyrin IX (ZnPP IX), a specific HO-1 inhibitor, dimanganese decacarbonyl (DMDC), a CO donor, or an equivalent volume of their respective vehicles were injected subcutaneously 30 min prior to local challenge with toxin A (25 or 50 µg per ileal loop) or PBS. Intestinal ileal loop weight/length ratios were calculated 3 h later. Ileal tissues were collected for histological analysis and measurement of myeloperoxidase (MPO) activity, tumour necrosis factor alpha (TNF-α) and interleukin-1 beta (IL-1ß) production by ELISA and immunohistochemistry for HO-1. Treatment of mice subjected to C. difficile toxin A (TcdA) with haemin or DMDC prevented oedema, mucosal disruption and neutrophil infiltration observed in histological analysis. It also decreased TcdA-induced MPO activity and TNF-α or IL-1ß production. In contrast, the specific HO-1 inhibitor (ZnPP IX) exacerbated all these evaluated parameters. TcdA increased HO-1 expression as seen by immunohistochemistry. These results suggest that the HO-1/CO pathway exerts a protective role in TcdA-induced enteritis and that its pharmacological modulation might be important for the management of C. difficile-associated disease.

Clinical outcome after acute ischaemic stroke: the influence of restless legs syndrome.

BACKGROUND AND PURPOSE: the objective was to evaluate the presence of Restless Legs Syndrome (RLS) in acute stroke, its association with sleep disturbances and clinical outcome during long-term follow-up. METHODS: this was a longitudinal study (N = 96, 59 men, mean age 64.0 ± 8.9) of cases with acute ischaemic stroke. Patients were asked about the occurrence of RLS symptoms before the cerebrovascular event. RLS was diagnosed using the criteria established by the International RLS Study Group. Stroke outcome was estimated by the Barthel Index and the modified Rankin Scale. Daytime somnolence (Epworth Sleepiness Scale -ESS > 10), poor sleep quality (Pittsburgh Sleep Quality Index -PSQI > 5) and risk of obstructive sleep apnea (OSA) (Berlin questionnaire) were evaluated. RESULTS: twelve patients (12.5%) met the diagnostic criteria for RLS. All cases had symptoms of RLS before stroke. However, none of the cases had a previous medical diagnosis of RLS or were on use of specific medication. In only one case, a family history of RLS was found. In all patients, RLS symptoms started after the age of 40 (mean age 64 ± 6.7). Daytime sleepiness (44.8%) and poor quality sleep (62.8%) were present. Patients with RLS (12.5%) presented greater neck circumference (P = 0.04) and worse sleep quality (P = 0.007). Risk of OSA (56.2%) was associated with hypertension [OR = 0.12; CI=0.03-0.42]. Stroke outcome was significantly worse at three and 12 months (ancova, P < 0.005) in patients with RLS, remaining after adjustment for diabetes and body mass index (P < 0.05). CONCLUSIONS: patients with acute stroke and RLS have worse clinical outcome, at three and 12 months of follow-up.

Effect of atorvastatin on 5-fluorouracil-induced experimental oral mucositis.

PURPOSE: Oral mucositis (OM) is a frequent side effect in patients with cancer. We investigate the effect of atorvastatin (ATV), a cholesterol-lowering drug, on OM induced by 5-fluorouracil (5-FU) in hamsters. METHODS: OM was induced by the i.p. administration of 5-FU, with excoriations of the cheek pouch mucosa. The animals were pretreated with i.p. ATV 1, 5 or 10 mg/kg or vehicle (saline and 5% (vol/vol) ethanol) 30 min before 5-FU injection and daily for 5 or 10 days. Samples of cheek pouches and main organs were removed for histopathological analysis, determination of TNF-α, IL-1ß, nitrite, non-protein sulfhydryl group (NP-SH) levels, myeloperoxidase (MPO) assay and immunohistochemistry for induced nitric oxide synthase (iNOS). Blood was collected for a leukogram analysis of biochemical parameters and analysis of bacteremia. RESULTS: ATV at doses of 1 and 5 mg/kg reduced mucosal damage and inflammation, as well as the levels of cytokines, nitrite and myeloperoxidase activity on the 5th and 10th day of OM and immunostaining for iNOS on the 5th day of OM.ATV at 1 mg/kg increased cheek pouch NP-SH when compared to 5-FU groups on the 10th day of OM. The association between ATV 5 mg/kg and 5-FU decreased the survival rate, amplified the leukopenia of animals, increased transaminase serum levels and caused liver lesions. We also detected the presence of Gram-negative bacillus in the blood of 100% of the animals treated with ATV 5 mg/kg + 5-FU. CONCLUSIONS: Atorvastatin prevented mucosal damage and inflammation associated with 5-FU-induced OM, but the association of a higher dose of ATV with 5-FU induced hepatotoxicity and amplified leukopenia.

Restless leg syndrome, sleep quality and fatigue in multiple sclerosis patients.

We have tested the hypothesis that restless leg syndrome (RLS) is related to quality of sleep, fatigue and clinical disability in multiple sclerosis (MS). The diagnosis of RLS used the four minimum criteria defined by the International Restless Legs Syndrome Study Group. Fatigue was assessed by the Fatigue Severity Scale (FSS >27), quality of sleep by the Pittsburgh Sleep Quality Index (PSQI >6), excessive daytime sleepiness by the Epworth Sleepiness Scale (ESS >10) and clinical disability by the Expanded Disability Status Scale (EDSS). Forty-four patients (32 women) aged 14 to 64 years (43 +/- 14) with disease from 0.4 to 23 years (6.7 +/- 5.9) were evaluated. Thirty-five were classified as relapsing-remitting, 5 as primary progressive and 4 as secondary progressive. EDSS varied from 0 to 8.0 (3.6 +/- 2.0). RLS was detected in 12 cases (27%). Patients with RLS presented greater disability (P = 0.01), poorer sleep (P = 0.02) and greater levels of fatigue (P = 0.03). Impaired sleep was present in 23 (52%) and excessive daytime sleepiness in 3 cases (6.8%). Fatigue was present in 32 subjects (73%) and was associated with clinical disability (P = 0.000) and sleep quality (P = 0.002). Age, gender, disease duration, MS pattern, excessive daytime sleepiness and the presence of upper motor neuron signs were not associated with the presence of RLS. Fatigue was best explained by clinical disability and poor sleep quality. Awareness of RLS among health care professionals may contribute to improvement in MS management.

Restless leg syndrome, sleep quality and fatigue in multiple sclerosis patients

We have tested the hypothesis that restless leg syndrome (RLS) is related to quality of sleep, fatigue and clinical disability in multiple sclerosis (MS). The diagnosis of RLS used the four minimum criteria defined by the International Restless Legs Syndrome Study Group. Fatigue was assessed by the Fatigue Severity Scale (FSS >27), quality of sleep by the Pittsburgh Sleep Quality Index (PSQI >6), excessive daytime sleepiness by the Epworth Sleepiness Scale (ESS >10) and clinical disability by the Expanded Disability Status Scale (EDSS). Forty-four patients (32 women) aged 14 to 64 years (43 ± 14) with disease from 0.4 to 23 years (6.7 ± 5.9) were evaluated. Thirty-five were classified as relapsing-remitting, 5 as primary progressive and 4 as secondary progressive. EDSS varied from 0 to 8.0 (3.6 ± 2.0). RLS was detected in 12 cases (27 percent). Patients with RLS presented greater disability (P = 0.01), poorer sleep (P = 0.02) and greater levels of fatigue (P = 0.03). Impaired sleep was present in 23 (52 percent) and excessive daytime sleepiness in 3 cases (6.8 percent). Fatigue was present in 32 subjects (73 percent) and was associated with clinical disability (P = 0.000) and sleep quality (P = 0.002). Age, gender, disease duration, MS pattern, excessive daytime sleepiness and the presence of upper motor neuron signs were not associated with the presence of RLS. Fatigue was best explained by clinical disability and poor sleep quality. Awareness of RLS among health care professionals may contribute to improvement in MS management.

Ehlers-Danlos syndrome type IV and multiple aortic aneurysms--a case report.

Beside atherosclerosis, aortic aneurysms can be part of the clinical spectrum of many systemic diseases, including infectious, inflammatory, genetic and, less often, congenital disorders. A 48-year-old white man presented with multiple large aneurysms of the aorta and its main branches. Medical history was unremarkable except for the presence of a softened abdominal mass since he was 28 years old. On the physical examination, an arterial murmur was heard over the left carotid artery and a palpable mass was noted in the whole right side of the abdomen. No skin or joint abnormalities were noted. Aortography, computed tomography, and magnetic resonance angiography showed multiple large aneurysms of the descending thoracic and abdominal aorta. Aneurysms of the innominate, left subclavian, and carotid arteries were also seen. This case resembles those previously reported, in which multiple aortic aneurysms were associated with abnormalities of the type III procollagen gene (COL3A1). Although the classic stigmas of the Ehlers-Danlos syndrome type IV were lacking, this genetic disease may be the cause of the multiple aneurysms in this patient.