When residents die in Portuguese care facilities: The experiences and practices of care facility directors before and during the COVID-19 pandemic.

This qualitative exploratory study aimed to describe the experiences and practices of care facility directors regarding residents dying in Portuguese residential care facilities (RCFs) before and during the COVID-19 pandemic. Data was obtained from 17 care director facilities who participated in focus groups. The data analysis followed the interpretative phenomenological approach. Participants stated that practices surrounding the death of the residents in Portuguese RCFs changed significantly from before to during the pandemic, regarding both those who died from COVID-19 infection and from other conditions. Four themes emerged that illustrated the situation during the peaks of the pandemic. During the pandemic, practices and experiences quite different from the usual ones have emerged. These results support the importance of a good death, and dying with dignity in RCFs, and the need for policies, practices, and training on the death of residents in RCFs in Portugal.

Non-adherence to COVID-19 lockdown: who are they? A cross-sectional study in Portugal.

OBJECTIVES: The SARS-CoV-2 virus has spread worldwide, leading governments to implement mitigation measures. Understanding the reluctance to adhere to non-pharmacological interventions might help promote adherence to these measures. This study aimed to identify factors associated with non-adherence to the first lockdown in Portugal. STUDY DESIGN: Cross-sectional study. METHODS: This study used data from a Portuguese community-based survey entitled 'COVID-19 Barometer: Social Opinion'. Data were collected on risk perception, health status and social experiences using a snowball sampling technique. The event of interest corresponded to participants who reported not staying home during the lockdown period, serving as a proxy for non-adherence to lockdown. Logistic regression was used to identify factors associated with non-adherence to the first lockdown. RESULTS: Responses from 133,601 individual questionnaires that were completed during the first week of the first lockdown in 2020 were analysed. A minority of participants (5.6%) reported non-adherence to lockdown (i.e. leaving home for reasons other than essential situations). Working in the workplace was the factor with the strongest association of non-adherence to the lockdown. Several other factors were also associated with non-adherence to the first lockdown; namely, being a man, being a student, having a low level of education, having a low income, living alone or with a high-infection-risk professional (e.g. doctor, nurse, pharmaceutical, health technician, firefighter, police officer, military, essential services worker), perceiving the risk of getting COVID-19 to be high, not having social support in case of infection, feeling agitated, sad or anxious every day, and considering the preventive measures to be unimportant or inadequate. CONCLUSIONS: Non-adherence to lockdown was associated with socio-economic, trust and perception factors. Future research should investigate the mechanisms underlying these associations to help identify the population groups who are most at risk of non-adherence.

Tuberculosis infection in HIV vs. non-HIV patients.

OBJECTIVES: Tuberculosis (TB) is the most common opportunistic infection and cause of mortality among people living with HIV, and it is possible that it may also influence the evolution of the HIV infection. We assessed the differences between HIV-positive and -negative people infected with TB. METHODS: The present study is a cross-sectional retrospective study by electronic record revision. We included patients admitted to a tertiary hospital with a diagnosis of TB between 2011 and 2016, comparing those with HIV coinfection with non-HIV patients, according to demographic and clinical characteristics. RESULTS: This study included 591 patients, of whom 32% were HIV-coinfected. HIV-TB patients were younger, with a predominance of male gender. Considering TB risk factors, there was a higher prevalence of homelessness and intravenous drug use in the HIV group. In the non-HIV group, direct contact with other patients with TB and immunosuppression were more prevalent. Relative to TB characteristics, the HIV-coinfected group presents with a higher prevalence of disseminated disease and a higher occurrence of previous TB infection. Cancer was the most frequent cause of immunosuppression in the HIV group and the number testing positive for TB via microbiological culture was lower. Assessment of microbiological resistance and in-hospital mortality showed similar numbers in both groups. CONCLUSIONS: There are few papers comparing clinical course of TB between HIV-infected and non-infected patients. Our study differs from others in the literature as we focused on a country with middling incidence of TB and further characterized the differences between HIV-infected and non-infected patients which can contribute to the management of these patients.

Validation of an in vitro system to trigger changes in the gene expression of effectors of Sclerotinia sclerotiorum.

AIMS: Sclerotinia sclerotiorum, the causal agent of white mold, can infect several host species, including economically important crops. In this study, we propose and validate a new in vitro system able to mimic the conditions of interaction with the host and promote the induction of S. sclerotiorum effectors. METHODS AND RESULTS: For culture media production, we selected three plant species, common bean (Phaseolus vulgaris L, cv. Requinte.), maize (Zea mays, cv. BRS1030) and beggarticks (Bidens pilosa). To validate this system as an in vitro inducer of effectors, the qRT-PCR technique was used to investigate the expression profile of some S. sclerotiorum effector genes in each growth medium at different times after inoculation. CONCLUSION: The results obtained in this study provide a validation of a new method to study S. sclerotiorum during mimetic interaction with different hosts. Although leaf extract does not fully represent the plant environment, the presence of plant components in the culture medium seems to induce effector genes, mimicking in planta conditions. The use of MEVM is simpler than in planta growth, bypasses problems such as the amount of mycelium produced, as well as contamination of host cells during transcriptomic and proteomic analyses. SIGNIFICANCE AND IMPACT OF THE STUDY: We have devised MEVM media as a model mimicking the interaction of S. sclerotiorum and its hosts and used it to evaluate in vitro expression of effectors normally expressed only in planta.

Trajectories of depressive symptoms through adolescence and young adulthood: social and health outcomes.

Depressive symptoms display heterogeneous trajectories across adolescence, which can lead to different consequences. This study aimed to identify trajectories of depressive symptoms from adolescence to young adulthood, assessing the association with social and health outcomes at young adulthood. Adolescents born in 1990, enrolled in schools of Porto, Portugal, in 2003-2004 (EPITeen study) completed the Beck Depression Inventory II (BDI-II) at 13, 17 and 21 years. Mixed models and model-based clustering were used to describe the trajectories in the BDI-II score (n = 2010). Outcomes were assessed at age 21 years with self-administered questionnaires and face-to-face interviews (n = 1594). Odds ratios or regression coefficients, with 95% confidence intervals, were estimated using logistic and linear regression. Three trajectory classes of depressive symptoms were identified, similar in shape in both sexes: High (8.4%), Moderate (31.3%) and Low (60.2%). Participants in High or Moderate classes were more likely to describe lower scores of community involvement, more medical appointments during the last year, higher levels of pain and had higher probability of self-rating health as "good" or "fair or poor". Females in the High and Moderate classes were more likely to be current smokers, to describe ever using drugs and to report more sexual partners, emergency room visits and the use of antidepressants. The risk of depressive symptoms in adulthood is likely to be early determined in adolescence. The trajectory classes with higher levels of symptoms were associated with worse social and health outcomes.

Primary surgical management of anterior pelvic organ prolapse: a systematic review, network meta-analysis and cost-effectiveness analysis.

BACKGROUND: Anterior compartment prolapse is the most common pelvic organ prolapse (POP) with a range of surgical treatment options available. OBJECTIVES: To compare the clinical effectiveness and cost-effectiveness of surgical treatments for the repair of anterior POP. METHODS: We conducted a systematic review of randomised controlled trials comparing surgical treatments for women with POP. Network meta-analysis was possible for anterior POP, same-site recurrence outcome. A Markov model was used to compare the cost-utility of surgical treatments for the primary repair of anterior POP from a UK National Health Service perspective. MAIN RESULTS: We identified 27 eligible trials for the network meta-analysis involving eight surgical treatments tested on 3194 women. Synthetic mesh was the most effective in preventing recurrence at the same site. There was no evidence to suggest a difference between synthetic non-absorbable mesh, synthetic partially absorbable mesh, and biological mesh. The cost-utility analysis, which incorporated effectiveness, complications and cost data, found non-mesh repair to have the highest probability of being cost-effective. The conclusions were robust to model inputs including effectiveness, costs and utility values. CONCLUSIONS: Anterior colporrhaphy augmented with mesh appeared to be cost-ineffective in women requiring primary repair of anterior POP. There is a need for further research on long-term effectiveness and the safety of mesh products to establish their relative cost-effectiveness with a greater certainty. TWEETABLE ABSTRACT: New study finds mesh cost-ineffective in women with anterior pelvic organ prolapse.

Factors related to condomless anal intercourse between men who have sex with men: results from a European bio-behavioural survey.

BACKGROUND: Relationship status is an important factor associated with condomless anal intercourse (CAI) amongst men who have sex with men (MSM). METHODS: A multi-centre bio-behavioural survey with MSM was conducted in 13 European cities (n = 4901) exploring factors associated with CAI via bivariate and multivariate multilevel logistic regression analyses. RESULTS: Likelihood of CAI with casual partners was associated with being 'out' to a majority (AOR = 1.19;95% CI 1,1.42); knowing their HIV status (AOR = 1.86; 95% CI 1.25,2.76); using substances (1-2 AOR = 1.39; 95% CI 1.16,1.63, 2+ AOR = 1.81; 95% CI 1.35,2.42); being older (AOR = 0.98; 95% CI 0.97,0.99); successful sero-communication (AOR = 0.79; 95% CI 0.67,0.94); and, not having a recent HIV test (AOR = 0.78; 95% CI 0.66,0.92). CAI with steady partners was associated with successful sero-communication (AOR = 2.72; 95% CI 2.72,3.66); not having a recent HIV test (AOR = 1.26; 95% CI 1.09,1.46), and; being older (AOR = 0.99; 95% CI 0.98,0.99). CONCLUSIONS: Understandings of partner type and/or relationship status in relation to CAI amongst MSM can potentially play an important role in the development of culturally appropriate HIV/STI prevention and risk-reduction efforts targeting at-risk MSM. Our results speak to the need to consider segmented and tailored public health and health promotion initiatives for MSM with differing CAI behaviours and relationship profiles.

Depressive symptoms at 13 years as predictors of depression in older adolescents: a prospective 4-year follow-up study in a nonclinical population.

Depression is the most important source of disability in adolescents, partially due to its recurrence. There is a lack of studies on population-based samples investigating the continuity of depressive symptoms during adolescence. This study evaluates depressive symptoms at early adolescence as predictors of depressive symptoms later in adolescence. Urban adolescents born in 1990 and enrolled in schools of Porto, Portugal, in 2003-2004 (EPITeen study) were evaluated at 13 and 17 years (n = 1106, 55.9% females), and completed a questionnaire comprising health behaviors and Beck Depression Inventory II (BDI-II) to assess depressive symptoms. A questionnaire on socio-demographic and clinical characteristics was self-reported. Regression coefficients (ß) and 95% confidence intervals (CI) were estimated using simple linear regression. The prevalence of adolescents with depressive symptoms above the cut-off (BDI-II > 13) was 11.9% at 13 years (girls: 17.1%; boys: 5.3%) and 10.8% at 17 years (girls: 14.7%; boys: 5.7%). Almost 6% of girls and 2% of boys had BDI-II > 13 at both assessments, and 35% of girls and boys with BDI-II > 13 at baseline also had BDI-II > 13 at follow-up. For both genders, depressive symptoms at age 13 were independently associated with depressive symptoms at age 17 (girls: ß = 0.35, 95% CI 0.28-0.42; boys: ß = 0.37, 95% CI 0.30-0.44). Depressive symptoms at age 13 were an independent predictive factor for adolescents' depressive symptoms at age 17. The prevalence of adolescents with BDI-II > 13 was higher in females, but the strength of this association was similar in both genders, highlighting the heavy burden of depressive symptoms already at an early age, among girls and boys.

Fusion of plectasin derivative NZ2114 with hydrophilic random coil polypeptide: Recombinant production in Pichia pastoris and antimicrobial activity against clinical strain MRSA.

One of the roadblocks towards the practical use of antimicrobial peptides for medical use is their relatively high cost when synthesized chemically. Effective recombinant production has only been successful in some cases, such as the previously reported production in Pichia pastoris of the antimicrobial plectasin derivative peptide NZ2114. The same production host has also been used extensively to produce so-called protein-polymers: sequences that consist of repetitions of simple amino acid motifs found in structural proteins such as collagen and elastin, and that can be designed to self-assemble in micelles, fibers and hydrogels. With the eventual goal of producing recombinant biomaterials such as antimicrobial protein polymer, we here explore the secreted production in Pichia pastoris of a fusion of NZ2114 with a hydrophilic random coil protein polymer CP4 . The intact NZ2114-CP4 fusion copolymer was produced with a yield of purified protein on the order of 1 g.L-1 supernatant. We find that purified NZ2114-CP4 has an activity against clinical strain MRSA, but very much lower than activity of chemically synthesized NZ2114. We conclude that possibly, the activity of NZ2114 is impaired by the C-terminal attachment to the protein polymer chain, but other reasons for the low activity cannot yet be excluded either. This article is protected by copyright. All rights reserved.

Human cysticercosis in Portugal: long gone or still contemporary?

Cysticercosis, a leading cause of acquired epilepsy in developing countries, has been controlled or eradicated in industrialized countries. This paradigm has recently been challenged, with human neurocysticercosis (NCC) being increasingly diagnosed in these countries. In order to assess the NCC burden in Portugal, a retrospective study on NCC hospitalizations (2006-2013) was conducted based on the national database on hospital morbidity: 357 hospitalized cases were detected. NCC was most frequent in the following age groups: 20-64 years (n = 197, 55·2%) >64 years (n = 111, 31·1%), and <20 years (n = 49, 13·7%). In the Norte and Centro regions cases tended to be older than in the Lisboa and Vale do Tejo Region. The results raise concerns for imported and autochthonous disease, suggesting the Lisboa and Vale do Tejo Region, due to its higher frequency of cases at younger ages, as a priority for research and intervention, and further suggest that NCC should be under surveillance (notifiable). The National Observatory of Cysticercosis and Taeniasis has been established and will define NCC cases as well as monitoring and surveillance.

Vasomotor symptoms resulting from natural menopause: a systematic review and network meta-analysis of treatment effects from the National Institute for Health and Care Excellence guideline on menopause.

BACKGROUND: Vasomotor symptoms (VMSs) are the hallmarks of menopause, occurring in approximately 75% of postmenopausal women in the UK, and are severe in 25%. OBJECTIVES: To identify which treatments are most clinically effective for the relief of VMSs for women in natural menopause without hysterectomy. SEARCH STRATEGY: English publications in MEDLINE, Embase, and The Cochrane Library up to 13 January 2015 were searched. SELECTION CRITERIA: Randomised controlled trials (RCTs) of treatments for women with a uterus for the outcomes of frequency of VMSs (up to 26 weeks), vaginal bleeding, and discontinuation. DATA COLLECTION AND ANALYSIS: Bayesian network meta-analysis (NMA) using mean ratios (MRs) and odd ratios (ORs). MAIN RESULTS: Across the three networks, 47 RCTs of 16 treatment classes (n = 8326 women) were included. When compared with placebo, transdermal estradiol and progestogen (O+P) had the highest probability of being the most effective treatment for VMS relief (69.8%; MR 0.23; 95% credible interval, 95% CrI 0.09-0.57), whereas oral O+P was ranked lower than transdermal O+P, although oral and transdermal O+P were no different for this outcome (MR 2.23; 95% CrI 0.7-7.1). Isoflavones and black cohosh were more effective than placebo, although not significantly better than O+P. Not only were selective serotonin reuptake inhibitors (SSRIs) or serotonin-norepinephrine reuptake inhibitors (SNRIs) found to be ineffective in relieving VMSs, but they also had significantly higher odds of discontinuation than placebo. Limited data were available for bleeding, therefore no conclusions could be made. CONCLUSION: For women who have not undergone hysterectomy, transdermal O+P was the most effective treatment for VMS relief. TWEETABLE ABSTRACT: Which treatment best relieves menopause flushes? Results from the #NICE guideline network meta-analysis.

Cognitive impairment and magnetic resonance imaging correlates in primary progressive multiple sclerosis.

OBJECTIVES: To characterize cognitive impairment in primary progressive multiple sclerosis (PPMS) and to correlate the pattern of cognitive deficits with brain magnetic resonance imaging (MRI) volumetric data. MATERIALS AND METHODS: In a multicenter cross-sectional study, we recruited consecutive patients with PPMS as well as age, sex, and education level-matched healthy controls (HC). All participants underwent neuropsychological (NP) assessment, and brain MRI was performed in patients with PPMS for analysis of lesion load, subcortical GM volumes, and regional cortical volumes. RESULTS: We recruited 55 patients with PPMS and 36 HC. Thirty-six patients were included in the MRI analysis. Patients with PPMS performed significantly worse than HC in all NP tests. Subcortical GM volume was significantly correlated with all NP tests, except for Stroop Test, with the largest effect for the thalamus (r=-.516 [BVMT-R DR, P=.016 FDR-corrected] to r=.664 [SDMT, P<.001 FDR-corrected]). In the stepwise linear regression model, thalamic volume was the only predictor of performance in all NP tests. CONCLUSION: Cognitive impairment is common in PPMS and affects all evaluated cognitive domains. Subcortical GM volume, particularly of the thalamus, is a strong predictor of cognitive performance, suggesting it has a central role in the pathophysiology of PPMS-related cognitive dysfunction.

Purification of plant complex protein extracts in non-denaturing conditions by in-solution isoelectric focusing.

An alternative approach for plant complex protein extracts pre-purification by in-solution isoelectric focusing in non-denaturing conditions is presented. The separation of biologically active proteins, in narrow ranges of isoelectric point (pI) was obtained by a modified OFFGEL electrophoresis. Two different water-soluble protein extracts from Phragmites leaves were fractionated into 24 fractions within a 3-10 pI range at 10 °C in the absence of denaturing/reducing agents. One-dimensional electrophoretic analysis revealed different protein distribution patterns and the effective fractionation of both protein extracts. Peroxidase activity of each fraction confirmed that proteins remained active and pre-purification occurred. Biological triplicates assured the needed reproducibility.

Methods to induce labour: a systematic review, network meta-analysis and cost-effectiveness analysis.

OBJECTIVES: To compare the clinical effectiveness and cost-effectiveness of labour induction methods. METHODS: We conducted a systematic review of randomised trials comparing interventions for third-trimester labour induction (search date: March 2014). Network meta-analysis was possible for six of nine prespecified key outcomes: vaginal delivery within 24 hours (VD24), caesarean section, uterine hyperstimulation, neonatal intensive care unit (NICU) admissions, instrumental delivery and infant Apgar scores. We developed a decision-tree model from a UK NHS perspective and calculated incremental cost-effectiveness ratios, expected costs, utilities and net benefit, and cost-effectiveness acceptability curves. MAIN RESULTS: In all, 611 studies comparing 31 active interventions were included. Intravenous oxytocin with amniotomy and vaginal misoprostol (≥50 µg) were most likely to achieve VD24. Titrated low-dose oral misoprostol achieved the lowest odds of caesarean section, but there was considerable uncertainty in ranking estimates. Vaginal (≥50 µg) and buccal/sublingual misoprostol were most likely to increase uterine hyperstimulation with high uncertainty in ranking estimates. Compared with placebo, extra-amniotic prostaglandin E2 reduced NICU admissions. There were insufficient data to conduct analyses for maternal and neonatal mortality and serious morbidity or maternal satisfaction. Conclusions were robust after exclusion of studies at high risk of bias. Due to poor reporting of VD24, the cost-effectiveness analysis compared a subset of 20 interventions. There was considerable uncertainty in estimates, but buccal/sublingual and titrated (low-dose) misoprostol showed the highest probability of being most cost-effective. CONCLUSIONS: Future trials should be designed and powered to detect a method that is more cost-effective than low-dose titrated oral misoprostol. TWEETABLE ABSTRACT: New study ranks methods to induce labour in pregnant women on effectiveness and cost.

Fish oils against Burkholderia and Pseudomonas aeruginosa: in vitro efficacy and their therapeutic and prophylactic effects on infected Galleria mellonella larvae.

AIM: This study investigates the antimicrobial effects of fish oil-based formulas rich in omega-3 fatty acids (free fatty acids, ethyl esters or triacylglycerols), against cystic fibrosis (CF) pathogens (Burkholderia cenocepacia K56-2 and Pseudomonas aeruginosa PAO1), often resistant to multiple antibiotics. METHODS AND RESULTS: The fish oils have shown antibacterial efficacy, although activity was highest for the one containing the fatty acid EPA (eicosapentaenoic acid) and DHA (docosahexaenoic acid) in their free form (MIC value is 1·87% v/v for both pathogens). To test whether the fish oils could have a therapeutic and prophylactic potential in vivo, we assessed its efficacy using a Galleria mellonella caterpillar model of infection. The treatment of infected larvae with a single dose (7 h post infection) enhances the survival of larvae, being more pronounced with the free fatty acid form (EPAX 6000 FA). Moreover, we observed that the prophylactic food provision of the fish oil EPAX 6000 FA during 12 days prior to bacterial infection extended the life of the infected larvae. CONCLUSION: The fish oils, particularly in the free fatty acid form, are active in killing Burkholderia and Ps. aeruginosa. SIGNIFICANCE AND IMPACT OF THE STUDY: The possibility of using fish oils for the treatment of bacterial infections in CF patients.

A Bayesian framework to account for uncertainty due to missing binary outcome data in pairwise meta-analysis.

Missing outcome data are a common threat to the validity of the results from randomised controlled trials (RCTs), which, if not analysed appropriately, can lead to misleading treatment effect estimates. Studies with missing outcome data also threaten the validity of any meta-analysis that includes them. A conceptually simple Bayesian framework is proposed, to account for uncertainty due to missing binary outcome data in meta-analysis. A pattern-mixture model is fitted, which allows the incorporation of prior information on a parameter describing the missingness mechanism. We describe several alternative parameterisations, with the simplest being a prior on the probability of an event in the missing individuals. We describe a series of structural assumptions that can be made concerning the missingness parameters. We use some artificial data scenarios to demonstrate the ability of the model to produce a bias-adjusted estimate of treatment effect that accounts for uncertainty. A meta-analysis of haloperidol versus placebo for schizophrenia is used to illustrate the model. We end with a discussion of elicitation of priors, issues with poor reporting and potential extensions of the framework. Our framework allows one to make the best use of evidence produced from RCTs with missing outcome data in a meta-analysis, accounts for any uncertainty induced by missing data and fits easily into a wider evidence synthesis framework for medical decision making.

Ki-67 antigen expression in the mammary epithelium of female rats in persistent estrus treated with raloxifene.

OBJECTIVE: To evaluate Ki-67 antigen expression in the mammary epithelium of female rats in persistent estrus treated with raloxifene. MATERIALS AND METHODS: Forty-one Wistar-Hannover rats in persistent estrus induced by 1.25 mg of testosterone propionate were randomly divided into two groups: Group A (control, n = 21) in which the animals received only the vehicle (propylene glycol) and Group B (experimental, n = 20) in which the rats received 750 µg/day of raloxifene by gavage. After 21 days of treatment, all the animals were sacrificed and the first pair of abdominal-inguinal mammary glands was extirpated and fixed in 10% buffered formalin to investigate Ki-67 expression by immunohistochemistry. The data were analysed using Student's t-test (p < 0.05). RESULTS: The percentage of Ki-67-stained nuclei per 500 cells in the mammary epithelium was 42.33 ± 6.18 and 15.51 ± 3.71 [mean ± standard error of the mean (SEM)] in the control and experimental groups, respectively (p < 0.001). CONCLUSION: Raloxifene treatment significantly reduced Ki-67 expression in the mammary epithelium of rats in persistent estrus.

Similar hypotensive responses to resistance exercise with and without blood flow restriction.

Low intensity resistance exercise (RE) with blood flow restriction (BFR) has gained attention in the literature due to the beneficial effects on functional and morphological variables, similar to those observed during traditional RE without BFR, while the effects of BFR on post-exercise hypotension remain unclear. The aim of the present study was to compare the blood pressure (BP) response of trained normotensive individuals to RE with and without BFR. In this cross-over randomized trial, eight male subjects (23.8 ± 4 years, 74 ± 3 kg, 174 ± 4 cm) completed two exercise protocols: traditional RE (3 x 10 repetitions at 70% one-repetition maximum [1-RM]) and low intensity RE (3 x 15 repetitions at 20% 1-RM) with BFR. Blood pressure measurements were performed after 15 min of seated rest (0), immediately after and 10 min, 20 min, 30 min, 40 min, 50 min and 60 min after the experimental sessions. Similar hypotensive effects for systolic BP (SBP) were observed for both protocols (P < 0.05) after exercise, with no differences between groups (P > 0.05) and no statistically significant difference for diastolic BP (P > 0.05). These results suggest that in normotensive trained individuals, both traditional RE and RE with BFR induce hypotension for SBP, which is important to prevent cardiovascular disturbances.

A Response Surface Methodology study on the role of factors affecting growth and volatile phenol production by Brettanomyces bruxellensis ISA 2211 in wine.

The present study was aimed at determining the effect of glucose, ethanol and sulphur dioxide on the growth and volatile phenol production by Brettanomyces bruxellensis in red wines using a response surface methodology approach. Sulphur dioxide proved to have a significant (p < 0.05) negative linear and quadratic effect on growth and 4-ethylphenol production. Concentrations of sulphur dioxide higher than 20 mg L(-1), at pH 3.50, induced immediate loss of cell culturability under growth permissive levels of ethanol. Under high ethanol concentrations (14% v/v), the lag phase increased from 3 to 10 days, growth being fully arrested at 15% (v/v). Glucose up to 10 g L(-1) was found to be a significant factor (quadratic level) in biomass increase under low ethanol (<12.5% v/v) and low sulphite concentrations. However, when cells were inactivated by sulphur dioxide and ethanol, glucose (up to 10 g L(-1)) did not prevent cell death. Production of more than 50 µg L(-1) day(-1) of 4-ethylphenol was only observed in the presence of high numbers (10(6) CFU mL(-1)) of culturable cells, being stimulated by increasing glucose concentrations.