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Background Several studies reported an increased rate of indeterminate QuantiFERON-TB Gold Plus (QFT-P) assay results in patients with severe Coronavirus Disease (COVID)-19. Methods Aim of the study was to longitudinally evaluate QFT-P responses in patients who survived COVID-19, with a previous indeterminate result. Results We observed 223 patients with an indeterminate QFT-P assay among 949 patients hospitalized because of COVID-19 (23,5%) during 2020 and 2021. 36 patients among those with an indeterminate QFT-P assay were enrolled for reassessing the test. In 12 patients peripheral blood lymphocyte subsets were also reassessed. Considering disease severity, 30 were classified as severe and 6 as non-severe. Median age was 57,5 (interquartile range [IQR]: 49,5-63,8), with a prevalence of male sex (M/F: 24/12); median Charlson Comorbidity Index was 2 (IQR: 1-3). The second QFT-P assay was performed after at least 1 month from the first assay (median time 7 months, IQR: 5-12 months). All QFT-P assays gave a determined result: 2 positive (5.5%) and 34 negatives (94,4%). A statistically significant difference was observed after comparing the laboratory parameters at the time of the first and the second QFT-P assay: the absolute counts of total lymphocyte, total CD3+, CD4+ and CD8+ T-lymphocytes were significantly increased (p< 0.001) while neutrophil absolute counts, neutrophil to lymphocyte (N/L) ratio, D-dimer, fibrinogen, ferritin, C-reactive protein (CRP) were significantly reduced (p< 0.0001). Concerning the QFT-P assay, interferon gamma (INF-γ) production in the Mitogen-Nil, TB1-Nil and TB2-Nil conditions were significantly increased (p< 0.0001; p=0.0019; p=0.0205, respectively) (Table 1 and Figure 1). Conclusion Once the acute phase of COVID-19 is resolved, inflammatory markers and peripheral blood leucocyte counts tend to normalize with an effective INF-γ production after specific and nonspecific stimulation. All the 36 QFT-P showed a determinate result. Moreover, we observed 2 positive QFT-P assay, supporting the importance of retesting patients with indeterminate result to identify latent tuberculosis infection and monitor patients for possible reactivation because of the immune-suppression associated with COVID-19. Disclosures All Authors: No reported disclosures.

Background Patients undergoing haematopoietic stem cell transplantation (HSCT) have severe and prolonged immunodeficiency and the incidence of tuberculosis (TB) disease in those patients is 10 to 40 times higher than the general population Methods This retrospective study performed at Tor Vergata University Hospital in Rome (Italy) included all adult patients who underwent a HSCT in the Hematologic Transplant Unit from January 2015 to December 2019. Data on TB screening, performed through interferon-γ release assay (IGRA), were collected; patients were observed for TB reactivation in a follow up period of at least 3 years Results 323 HSCT recipients were enrolled in this study. Patients’ characteristics are summarized in table 1. 63 patients (19.5%) were not screened for TB, of which 6.3% were born overseas. 260 HSCT recipients (80.5%) were screened: 228 patients (87.7%) had a negative IGRA result, 21 (8.1%) tested positive, and 11 (4.2%) had an indeterminate result. 98.5% were screened before the HSCT. Of the 21 patients with a positive pre-HSCT IGRA test, 14 (66.7%) were males and significantly (p< 0.001) older (median age 60 [IQR] 58-65 years) than those with negative or indeterminate results. Almost all patients (95.2%) were Italian. Patients with a positive IGRA showed a higher monocyte count compared to the other groups (p=0.044), while those with indeterminate IGRA showed a tendence to lower lymphocyte absolute counts, compared to the other groups (p=0.060) (Table 2). All the 21 patients with a pre-HSCT positive IGRA test were radiologically and clinically screened for active TB and, once it was ruled out, treated for latent TB (LTBI) with isoniazid (INH) for 6 months. No events of toxicity or treatment interruption were reported. None of the patients was treated with rifampin. None of the 21 patients with pre-HSCT positive IGRA test developed active TB during the follow up period. Table 1.Characteristics of the study population, overall and divided according to IGRA testingTable 2.Characteristics of the IGRA tested patients, overall and divided by IGRA test results Conclusion To our knowledge, this is the first Italian study on the prevalence of LTBI in patients undergoing HSCT. 8.1% of the 260 patients screened with an IGRA tested positive. The great majority (95.2%) of LTBI patients were Italian and significantly older than IGRA-negative patients. All LTBI HSCT patients received INH treatment and no drug toxicity, nor TB reactivation was observed during the follow-up period Disclosures All Authors: No reported disclosures.

Background The extension of HIV reservoir in the rectal gut associated lymphoid tissue (GALT) and the ART influence on this compartment are still poorly studied. We aimed to evaluate the extent of HIV-DNA in GALT and peripheral blood of virologically suppressed patients (VS) Methods 40 HIV-1+ VS pts were enrolled. Blood and GALT HIV-DNA levels and residual plasma viremia (RV) were quantified by droplet digital-PCR. Rectal biopsies were collected during high resolution anoscopy. Lymphocyte (L) phenotype and antiretroviral drug concentrations were analysed in tissue and peripheral blood Results Pts median age (IQR) was 37 (32-45) yrs, 39 (97.5%) were males. At HIV-1 diagnosis, median (IQR) viremia and CD4+ were 4.7 (4.3-5) Log10cps/mL and 469 (299-627) cells/µL, respectively. 27 (67.5%) pts had HIV-1 B subtype. At the enrolment, median (IQR) CD4+ was 720 (583-1041) cells/µL and RV was 3 (2-6) cps/mL. All pts were on 2 NRTI-regimen with a 3rd drug (20 INSTI, 15 NNRTI, 5 PI). A positive correlation emerged among HIV-DNA levels in the GALT and PBMC (Rho= 0.432; p=0.005), CD8+ L counts (Rho=0.367; p=0.02), and a negative correlation emerged with blood CD3+ L percentage (Rho=-0.489; p=0.006). Peripheral blood HIV-DNA values were inversely correlated with the CD4+ nadir (Rho= -0.379; p=0.017) and blood CD4+/CD8+ ratio (Rho= -0.362; p=0.022), and directly correlated with GALT CD4+CD38+ (Rho= 0.414; p=0.028) and blood CD8+CD38+ L (%) (Rho= 0.400; p=0.026). A positive correlation among GALT and peripheral blood CD4+CD38+ (Rho= 0.555; p=0.005), CD8+CD38+ (Rho= 0.417; p=0.030), and CD19+CD38+ L (Rho= 0.498; p=0.007) was found. There was no difference in HIV-DNA levels in the GALT and PBMCs according to the ART 3rd drug. In a subgroup of 20 pts the analysis of rectal tissue and blood drug concentrations showed that tissue/plasma ratio was higher in pts on Elvitegravir (EVG) and Rilpivirine (RPV) (median ratio 2.8 and 1.7, respectively) GALT and Blood HIV-DNA levels Tissue-Plasma Drug concentration Conclusion HIV-DNA from rectal GALT is a valid marker to characterize viral reservoir. The correlation of higher GALT HIV-DNA levels, CD8+ T L counts and CD4 and CD8 activation levels suggests that a greater amount of virus in the anorectal reservoir can perpetuate the inflammatory state characteristic of HIV+ pts. EVG and RPV showed higher tissue exposure than other ART regimens Disclosures All Authors: No reported disclosures.

Background Long Covid Syndrome (LCS) is used to describe signs and symptoms that continue or develop after acute COVID–19 infection. Natural history and treatments of this syndrome are still poorly understood, even if evidence suggests the potential role of physical rehabilitation in improving symptoms in these subjects. Aim The aim of the present study was to evaluate safety, effectiveness and feasibility of a multidisciplinary, out–of–hospital, exercise–based cardiac rehabilitation (EBCR) program, of two months duration, in improving symptoms and physical and psychological parameters in patients with LCS Methods Thirty consecutive patients with LCS (18 males, mean age 58 years) underwent an accurate medical screening process including anthropometric and muscular strength evaluation, cardiopulmonary exercise test (CPET), quality of life (QoL) and psychological appraisal before and after an EBCR program. Results At baseline, all LCS patients were strongly symptomatic and showed severe impairments in physical performance, QoL and psychological parameters. No adverse effects and dropouts were observed during the exercise training sessions. After the EBCR program, COVID–19 residual symptoms significantly decreased and significant improvements in upper and lower limb muscular strength, CPET parameters, perceived physical and mental health, depression and anxiety were observed. Conclusions The present study confirms the severe physical and psychological impairment of patients with LCS and suggests that a multidisciplinary EBCR program could promote their physical and psychological recovery

Funding Acknowledgements Type of funding sources: None. Background Long Covid Syndrome (LCS) is used to describe signs and symptoms that continue or develop after acute COVID-19 infection. Natural history and treatments of this syndrome is still poorly understood. In literature there is currently a lack of data on the real effectiveness of a multidisciplinary rehabilitation program based on structured physical exercise (SPE) in these patients. Purpose To evaluate safety, effectiveness and feasibility of a structured individualized rehabilitation program in improving physical and psychological parameters in patients with LCS. Methods Twenty-eight patients with LCS (19 males, mean age 57 years) underwent an accurate medical screening process, body composition evaluation, cardiopulmonary exercise test (CPET), muscular strength assessment, quality of life (QoL), psychological assessment and counselling, before and after a 12-sessions SPE program. Results At baseline, all LCS patients showed severe impairments in physical performance, QoL and psychological parameters. No adverse effects and dropouts were observed during training session. After the rehabilitation program, significant improvement in CPET parameters, upper and lower limb muscular strength, perceived physical and mental health, body composition, depression and anxiety and Covid residual symptoms was observed. Conclusions The present study confirms severe impairment of patients with LCS and suggest that a multidisciplinary rehabilitation program based on SPE could promote their physical and psychological recovery.

Funding Acknowledgements Type of funding sources: None. Background Ischemic cerebral and cardiac events are the leading causes of mortality and morbidity in Minor non-Disabling Stroke (MiNDS) patients. Control of cardiovascular risk factors, including physical activity levels, is a key strategy in secondary prevention of MiNDS. However, there is a gap between recommendations and real achieved physical activity levels in these patients. Clinical benefits obtained with cardiovascular rehabilitation in cardiac ischemic patients could be obtained with similar programs also in patients with MiNDS, given the common pathophysiological substrate of these two diseases. Purpose To evaluate safety, effectiveness and feasibility of a cardiovascular rehabilitation program based on structured physical exercise (SPE) in patients with MiNDS. Methods 39 MiNDS patients (26 males, mean age 66 years) underwent an accurate medical screening process, body composition evaluation, cardiopulmonary exercise test (CPET) and muscular strength assessment, before and after a 12-sessions in-hospital SPE program, and after 6- and 12-months follow-up during which they could continue their training schedule on their own. Results After in-hospital rehabilitation program, a significant improvement in CPET parameters, body composition and muscular strength was observed. These modifications persisted in the 32 patients who continued 6- and 12- months follow-up training, while regressed in the 7 patients who stopped training. Conclusions Results of the present study show that a SPE-based cardiovascular rehabilitation program provides clinical benefit in patients with MiNDS.

Background Ischemic cerebral and cardiac events are the leading causes of mortality and morbidity in Minor non–Disabling Stroke (MiNDS) patients. Control of cardiovascular risk factors, including physical activity levels, is a key strategy in secondary prevention of MiNDS. However, there is a gap between recommendations and real achieved physical activity levels in these patients. Clinical benefits obtained with cardiovascular rehabilitation in cardiac ischemic patients could be obtained with similar programs also in patients with MiNDS, given the common pathophysiological substrate of these two diseases Purpose To evaluate safety, effectiveness and feasibility of a cardiovascular rehabilitation program based on structured physical exercise (SPE) in patients with MiNDS Methods 39 MiNDS patients (26 males, mean age 66 years) underwent an accurate medical screening process, body composition evaluation, cardiopulmonary exercise test (CPET) and muscular strength assessment, before and after a 12–sessions in–hospital SPE program, and after 6– and 12–months follow–up during which they could continue their training schedule on their own Results After in–hospital rehabilitation program, a significant improvement in CPET parameters, body composition and muscular strength was observed. These modifications persisted in the 32 patients who continued 6– and 12– months follow–up training, while regressed in the 7 patients who stopped training Conclusions - Results of the present study show that a SPE–based cardiovascular rehabilitation program provides clinical benefit in patients with MiNDS

We describe the clinical case of a 48–year–old master athlete, runner (marathon runner), always asymptomatic at rest and during effort, with a family history of ischemic heart disease and no other known cardiovascular risk factors. On the occasion of the pre–partecipation screening visit, he presented an altered baseline electrocardiogram with low peripheral voltages and T wave inversion in V1–V5 leads and inferior leads. The echocardiogram documented morphofunctional alterations in the right ventricle and raises the suspicion of arrhythmogenic cardiomyopathy, later confirmed by cardiac magnetic resonance with Gadolinium. During effort, even repetitive premature ventricular beats LBBB with superior axis morphology appeared. Also, torsade de pointes episode with presyncopal sensation. He was hospitalized with a diagnosis of threatening arrhythmias in right ventricular arrhythmogenic heart disease. Coronary angiography was negative. We recommend beta–blocker therapy, genetic analysis, screening of family members. A subcutaneus defibrillator has been placed. The patient, against the advice of the Physicians, continued to train even at high intensity until he has a syncopal episode with correct intervention of the device on ventricular fibrillation. Since the patient has requested specific indications to continue training independently, an exercise test in beta–blocker therapy is repeated and the training thresholds are calculated. The case of this master athlete highlights the issue of the difficulty of screening the master athlete with frequent omission of symptoms (the patient already had syncopal/presyncopal episodes at home which he had omitted), poor adherence to therapeutic indications, possible negative effects of high intensity physical training on the evolution of arrhythmogenic cardiomyopathy.

Background Patients with a CRT–D devices represent an emerging population of subjects with chronic heart failure (CHF) who, according the current guidelines, must be referred to a cardiovascular rehabilitation (CR) program. Aims. The aims of the study were to evaluate in CRT–D recipients the feasibility, safety and efficacy of a physical exercise program, in a outpatient CR setting, to evaluate the need for an individual optimization of device programming. Methods Twenty–six CRT–D recipients underwent clinical and functional investigations, comprehensive of symptom–limited cardiopulmonary exercise testing, during which device programming were evaluated in order to obtain an adequate increase in heart rate and to maintain resynchronization therapy during exercise. They underwent a program of physician–supervised sessions of exercise training consisting of three 90 min visit/week. Results Baseline patients characteristics: male sex 77%; age 69±10 years; non–ischaemic cardiomyopathy 65%; ejection fraction 38±7 %; atrial fibrillation 12%. No adverse events occurred during training. The peak oxygen uptake has been significantly increased (from 12,6±6 to 17±9ml/kg/min). During cardiopulmonary exercise testing “Brady” functions has been reprogrammed in 10 patients (38%); no modifications to the "tachy" functions of the device were necessary. Conclusions Cardiac rehabilitation in CRT–D recipients is safe and effective. Device brady programming during exercise may be warranted in selected patients.

Background Little is known as yet about the outcome of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in children being treated for cancer. Methods We collected information on the clinical characteristics and outcomes of a cohort of 29 children (16 female and 13 male; median age, 7 years [range, 0–16 years]) diagnosed with SARS-CoV-2 infection while on chemotherapy/immunotherapy (n = 26), or after stem cell transplantation (n = 3) during the peak of the epidemic in Italy. These patients suffered from leukemia (n = 16), lymphoma (n = 3), solid tumors (n = 10), and Langerhans cell histiocytosis (n = 1). Results The course of the disease was mild in all cases, with only 12 children developing symptoms (pneumonia in 3 cases), and none needing intensive care. Fifteen patients were hospitalized, including 7 asymptomatic patients. Nine patients (including 5 with no symptoms) were given hydroxychloroquine, and 3 of them were also given lopinavir/ritonavir. Among the 26 patients on chemotherapy/immunotherapy, the treatment was suspended in 16 cases for a median of 26 days (range, 15–68 days), whereas 8 patients continued their chemotherapy and 2 had minor modifications to their treatment regimen. Conclusions SARS-CoV-2 infection seems to take a milder clinical course in children than in adults with cancer. Specific SARS-CoV-2 treatment seems unnecessary for most children. In light of our findings, and albeit with the necessary caution, we suggest avoiding major changes to planned anticancer treatments in pediatric patients acquiring COVID-19., We describe clinical characteristics and outcomes of 29 SARS-CoV-2–infected children with cancer. In comparison to adult oncology patients, children presented a milder clinical course. A substantial number of them continued their anticancer treatment without major modifications.

The use of thermotolerant yeast strains is an important attribute for a cost-effective high temperature biofermentation processes. However, the availability of thermotolerant yeast strains remains a major challenge. Isolation of temperature resistant strains from extreme environments or the improvements of current strains are two major strategies known to date. We hypothesised that bacteria are potential “hurdles” in the life cycle of yeasts, which could influence the evolution of extreme phenotypes, such as thermotolerance. We subjected a wild-type yeast, Lachancea thermotolerans to six species of bacteria sequentially for several generations. After coevolution, we observed that three replicate lines of yeasts grown in the presence of bacteria grew up to 37 °C whereas the controls run in parallel without bacteria could only grow poorly at 35 °C retaining the ancestral mesophilic trait. In addition to improvement of thermotolerance, our results show that the fermentative ability was also elevated, making the strains more ideal for the alcoholic fermentation process because the overall productivity and ethanol titers per unit volume of substrate consumed during the fermentation process was increased. Our unique method is attractive for the development of thermotolerant strains or to augment the available strain development approaches for high temperature industrial biofermentation.

Data on the accumulation of metals in wild edible and inedible macrofungi of the Boletaceae family are discussed here and compared with data reported in literature from northern Italy and other European countries, as well as from China, Brazil, and Africa. The results show a significant difference in the values detected in Sicilian fungi compared to those reported in the literature.

Background Paramyxoviruses include respiratory syncytial virus (RSV), parainfluenza virus (PIV), and human metapneumovirus (MPV), which may cause significant respiratory tract infectious disease (RTID) and mortality after allogeneic hematopoietic cell transplantation (HCT). However, clinical data regarding frequency and outcome are scarce. Methods We identified all paramyxovirus RTIDs in allogeneic HCT recipients diagnosed by multiplex polymerase chain reaction between 2010 and 2014. Baseline characteristics of patients, treatment, and outcome of each episode were analyzed; ie, moderate, severe, and very severe immunodeficiency (verySID) according to HCT ≤6 months, T- or B-cell depletion ≤3 months, graft-versus-host disease, neutropenia, lymphopenia, or hypo-gammaglobulinemia. Results One hundred three RTID episodes in 66 patients were identified (PIV 47% [48 of 103], RSV 32% [33 of 103], MPV 21% [22 of 103]). Episodes occurred in 85% (87 of 103) at >100 days post-HCT. Lower RTID accounted for 36% (37 of 103). Thirty-nine percent (40 of 103) of RTID episodes required hospitalization and more frequently affected patients with lower RTID. Six percent progressed from upper to lower RTID. Overall mortality was 6% and did not differ between paramyxoviruses. Sixty-one percent (63 of 103) of episodes occurred in patients with SID, and 20.2% (19 of 63) of episodes occurred in patients with verySID. Oral ribavirin plus intravenous immunoglobulin was administered in 38% (39 of 103) of RTIDs, preferably for RSV or MPV (P ≤ .001) and for SID patients (P = .001). Patients with verySID frequently progressed to lower RTID (P = .075), required intensive care unit transfer, and showed higher mortality. Conclusion Paramyxovirus RTID remains a major concern in allogeneic HCT patients fulfilling SID and verySID, emphasizing that efficacious and safe antiviral treatments are urgently needed.

Prostate cancer is the second most common cause of cancer and the sixth leading cause of cancer death among men worldwide. While localized prostate cancer can be cured, advanced and metastatic prostate cancer remains a significant therapeutic challenge. Malignant transformation is associated with important modifications of the cellular glycosylation profile, and it is postulated that these changes have a considerable relevance for tumor biology. Metastasis is a multiphasic process that encompasses angiogenesis, the spread of tumor cells and their growth at distant sites from the primary tumor location. Recognition of glycoconjugates by galectins, among other lectins, plays a fundamental role in the metastatic spread, tumor immune escape and the neovascularization process. Particularly in prostate cancer, both carbohydrates and galectins have been implicated in many cellular processes such as proliferation, apoptosis, migration and invasion. However, a limited number of studies assessed their potential implications in the induction of metastasis in prostate cancer patients or in animal models. Moreover, the role of galectin–glycan interactions in vivo still remains poorly understood; concerted effort should thus be made in order to shed some light on this question. This review summarizes current evidence on both the expression and role of glycans and galectins in prostate cancer, particularly turning our attention to the angiogenic and metastatic processes. Fil: Compagno, Daniel Georges. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina Fil: Gentilini, Lucas Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina Fil: Jaworski, Felipe Martín. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina Fil: Gonzales Perez, Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina Fil: Contrufo, Geraldine. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina Fil: Laderach, Diego Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina

Galectin-8 (gal-8) is a "tandem-repeat"-type galectin, containing two carbohydrate recognition domains connected by a linker peptide. gal-8 is expressed both in the cytoplasm and nucleus in vascular endothelial cells (ECs) from normal and tumor-associated blood vessels, and in lymphatic endothelial cells. Herein, we describe a novel role for gal-8 in the regulation of vascular and lymphatic angiogenesis and provide evidence of its critical implications in tumor biology. Functional assays revealed central roles for gal-8 in the control of capillary-tube formation, EC migration and in vivo angiogenesis. So far, two endothelial ligands have been described for gal-8, namely podoplanin in lymphatic vessels and CD166 (ALCAM, activated leukocyte cell adhesion molecule) in vascular ECs. Other related gal-8 functions are also summarized here, including cell adhesion and migration, which collectively demonstrate the multi-functionality of this complex lectin. Thus, gal-8 is an important component of the angiogenesis network, and an essential molecule in the extracellular matrix by providing molecular anchoring to this surrounding matrix. The implications of gal-8 in tumor angiogenesis remain to be further explored, but it is exciting to speculate that modulating gal-8-glycan interactions could be used to block lymphatic-vascular connections vital for metastasis. Fil: Troncoso, María Fernanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina Fil: Ferragut, Fatima Eneida del Valle. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina Fil: Bacigalupo, Maria Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina Fil: Cardenas Delgado, Victor Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina Fil: Nugnes, Lorena Gisela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina Fil: Gentilini, Lucas Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina Fil: Laderach, Diego Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina Fil: Wolfenstein, Carlota Elisa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina Fil: Compagno, Daniel Georges. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina Fil: Rabinovich, Gabriel Adrián. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina Fil: Elola, Maria Teresa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina

The origin of modern fruits brought to microbial communities an abundant source of rich food based on simple sugars. Yeasts, especially Saccharomyces cerevisiae, usually become the predominant group in these niches. One of the most prominent and unique features and likely a winning trait of these yeasts is their ability to rapidly convert sugars to ethanol at both anaerobic and aerobic conditions. Why, when, and how did yeasts remodel their carbon metabolism to be able to accumulate ethanol under aerobic conditions and at the expense of decreasing biomass production? We hereby review the recent data on the carbon metabolism in Saccharomycetaceae species and attempt to reconstruct the ancient environment, which could promote the evolution of alcoholic fermentation. We speculate that the first step toward the so-called fermentative lifestyle was the exploration of anaerobic niches resulting in an increased metabolic capacity to degrade sugar to ethanol. The strengthened glycolytic flow had in parallel a beneficial effect on the microbial competition outcome and later evolved as a “new” tool promoting the yeast competition ability under aerobic conditions. The basic aerobic alcoholic fermentation ability was subsequently “upgraded” in several lineages by evolving additional regulatory steps, such as glucose repression in the S. cerevisiae clade, to achieve a more precise metabolic control.

Yeast species belonging to the lineage that underwent the whole genome duplication (WGD), and including Saccharomyces cerevisiae, can grow under anaerobiosis and accumulate ethanol in the presence of glucose and oxygen. The pre-WGD yeasts, which branched from the S. cerevisiae lineage just prior to the WGD event, including Kluyveromyces lactis, are more dependent on oxygen and do not accumulate large amounts of ethanol in the presence of excess oxygen. Yeasts that belong to the so-called 'lower branches' of the yeast phylogenetic tree and diverged from S. cerevisiae more than 200 million years ago, have so far not been thoroughly investigated for their physiology and carbon metabolism. We have hereby studied several isolates of Candida albicans and Debaryomyces hansenii for their dependence on oxygen. C. albicans grew very poorly at oxygen concentration below 1 p.p.m. and D. hansenii could not grow at all. In aerobic batch cultivations C. albicans exhibited a predominately aerobic metabolism, accumulating only small amounts of ethanol (0.01-0.09 g g(-1) glucose). Apparently, C. albicans and several other pre-WGD yeasts still exhibit the original traits of the yeast progenitor: poor accumulation of ethanol under aerobic conditions and strong dependence on the presence of oxygen. (Less)

Background Extrapulmonary tuberculosis (EPTB) represents 25% of Worldwide tuberculosis and it is more commonly associated with immunodepression. The purpose of this study was to determinate the burden of EPTB in a low TB prevalence setting. Methods A retrospective evaluation of patients treated for TB at Tor Vergata Hospital and Terni Hospital (Italy) from January 2013 to November 2017 was done. Clinical charts, laboratory tests and radiological findings were reviewed and analysed. Data were elaborated using Yates’ method analysis, Fisher test and Pearson’s chi-square test. Results A total of 171 patients were enrolled from 2013 to 2017 in two Italian centers (Rome and Terni); 71% were males, with a mean age of 41.5 years. The number of TB diagnosis increased among the study period (6.6% in 2013 vs. 56% in 2017) and an increase of EPTB (23% in 2013 vs. 44% in 2017) was seen. Most commonly EPTB presented as generalized lymphadenitis (34%), osteomyelitis and spondylodiscitis (28%) and other sites localizations (31%). Statistical analysis revealed a significant correlation between geographical provenience and TB localization (P = 0.004). Extra European immigrants (76% Africans) resulted at higher risk of EPTB (OR 2.31; CI 95% 0.63–8.46), while being Caucasian showed a protective role toward EPTB development (P = 0.001). The risk of EPTB doubled in 2015–2017 respect 2013–2014. Conclusion From 2013 to 2017 an increase in TB admissions was documented with a significant higher number of EPTB cases, particularly in extra-European immigrants. The doubled risk in 2015–2017 was likely the consequence of the recent ongoing escalating levels of migration from African countries and may result as an emerging Public Health problem. Disclosures All authors: No reported disclosures.

Contamination of wine by Dekkera/Brettanomyces bruxellensis is mostly due to the production of off-flavours identified as vinyl- and especially ethyl-phenols, but these yeasts can also produce several other spoiling metabolites, such as acetic acid and biogenic amines. Little information is available about the correlation between growth, viability and off-flavour and biogenic amine production. In the present work, five strains of Dekkera bruxellensis isolated from wine were analysed over 3 months in wine-like environment for growth, cell survival, carbon source utilization and production of volatile phenols and biogenic amines. Our data indicate that the wine spoilage potential of D. bruxellensis is strain dependent, being strictly associated with the ability to grow under oenological conditions. 4-Ethyl-phenol and 4-ethyl-guaiacol production ranged between 0 and 2.7 and 2 mg L(-1), respectively, depending on the growth conditions. Putrescine, cadaverine and spermidine were the biogenic amines found.

Ebert, D. A., Compagno, L. J. V., and Cowley, P. D. 2008. Aspects of the reproductive biology of skates (Chondrichthyes: Rajiformes: Rajoidei) from southern Africa. – ICES Journal of Marine Science, 65: 81–102. New information is presented on the reproductive biology of 22 southern African skate species. Sex ratios for most species were relatively even. Sexual dimorphic differences in disc shape were evident in all species, but the total length (LT) to disc width (D) relationship was significantly different in only three species, and the LT to weight (W) relationship significant in just five species. Sexual dimorphism relative to maximum total length (LTmax) was absent in all but the two largest species. Males and females of the same species grow to a similar LTmax except those whose LTmax is >1.5 m LT. Size at first and 50% (LT50) sexual maturity was approximately the same for both sexes in all but the two largest species. First maturity occurred at >60% of LTmax for all species for which sufficient data were available, and most (n = 18) matured at >75% LTmax. The large size at maturity relative to LTmax suggests that growth slows or is partially suspended following sexual maturity. The egg cases of 15 species are described, and a key to their identification is presented. Egg cases in utero were observed throughout the year suggesting that most species reproduce year-round.

The optimisation and scale-up of a specific protein production process have to take into account cultivation conditions as well as cell physiology of growth and the influence of foreign protein expression on host cell metabolism. The ability of Zygosaccharomyces bailii to tolerate high sugar concentrations as well as high temperatures and acidic environments renders this “non-conventional” yeast suitable for the development of biotechnological processes like heterologous protein production. This work addresses the production of human interleukin-1β by a recombinant Z. bailii strain. We found that the heterologous protein production causes some modifications of the Z. bailii carbon metabolism, leading to a reduced biomass yield. The other important factor is the dependence of the recombinant IL-1β production/secretion on the growth rate. Among the cultivation strategies studied, the most appropriate in terms of production and productivity was the fed-batch mode.

Despite the importance of some Zygosaccharomyces species as agents causing spoilage of food, the carbon and energy metabolism of most of them is yet largely unknown. This is the case with Zygosaccharomyces bailii. In this study the occurrence of the Crabtree effect in the petite-negative yeast Z. bailii ATCC 36947 was investigated. In this yeast the aerobic ethanol production is strictly dependent on the carbon source utilised. In glucose-limited continuous cultures a very low level of ethanol was produced. In fructose-limited continuous cultures ethanol was produced at a higher level and its production increased with the dilution rate. As a consequence, on fructose the onset of respiro-fermentative metabolism caused a reduction in biomass yield. An immediate aerobic alcoholic fermentation in Z. bailii was observed during the transition from sugar limitation to sugar excess, both on glucose and on fructose. The analysis of some key enzymes of the fermentative metabolism showed a high level of acetyl-CoA synthetase in Z. bailii growing on fructose. At high dilution rates, the activities of glucose- and fructose-phosphorylating enzymes, as well as of pyruvate decarboxylase and alcohol dehydrogenase, were higher in cells during growth on fructose than on glucose.

Dekkera bruxellensis and Saccharomyces cerevisiae are considered two phylogenetically distant relatives, but they share several industrial relevant traits such as the ability to produce ethanol under aerobic conditions (Crabtree effect), high tolerance towards ethanol and acids, and ability to grow without oxygen. Beside a huge adaptability, D. bruxellensis exhibits a broader spectrum in utilization of carbon and nitrogen sources in comparison to S. cerevisiae. With the aim to better characterize its carbon source metabolism and regulation, the usage of galactose and the role that glucose plays on sugar metabolism were investigated in D. bruxellensis CBS 2499. The results indicate that in this yeast galactose is a non-fermentable carbon source, in contrast to S. cerevisiae that can ferment it. In particular, its metabolism is affected by the nitrogen source. Interestingly, D. bruxellensis CBS 2499 exhibits the 'short-term Crabtree effect', and the expression of genes involved in galactose utilization and in respiratory metabolism is repressed by glucose, similarly to what occurs in S. cerevisiae.

Introduction of the Lactobacillus casei lactate dehydrogenase (LDH) gene into Saccharomyces cerevisiae under the control of the TPI1 promoter yielded high LDH levels in batch and chemostat cultures. LDH expression did not affect the dilution rate above which respiro-fermentative metabolism occurred (Dc) in aerobic, glucose-limited chemostats. Above Dc, the LDH-expressing strain produced both ethanol and lactate, but its overall fermentation rate was the same as in wild-type cultures. Exposure of respiring, LDH-expressing cultures to glucose excess triggered simultaneous ethanol and lactate production. However, the specific glucose consumption rate was not affected, indicating that NADH reoxidation does not control glycolytic flux under these conditions.