Effects of Fermentation Temperature, Drying Temperature, Caliber Size, Starter Culture, and Sodium Lactate on Listeria monocytogenes Inactivation During Salami Production.

The effect of fermentation and drying temperatures, caliber, and sodium lactate on Listeria monocytogenes inactivation was studied in salami, produced in a pilot scale, inoculated with 107 CFU/g of Listeria innocua ATCC® 33090 as a surrogate microorganism for L. monocytogenes. Fermentation temperature varied between 24 and 30°C, drying temperature between 14 and 20°C, caliber between 5.1 and 13.2 cm, and sodium lactate initial concentrations in salamis were 0 and 2%. L. innocua counts, pH and water activity were determined in salamis over time. Sodium lactate (2%) decreased pH drop and Listeria inactivation during fermentation. Baranyi & Roberts equation was used to fit the experimental data and to estimate, for each test condition, inactivation rate (k), initial (Y0), and final counts of L. innocua (YEND). Total inactivation was calculated as Y0 minus YEND (Y0-YEND). Then, using a Box Benkhen experimental design, a quadratic model for k and a two-factor interaction model (2FI) for Y0 - YEND were obtained as functions of fermentation temperature, drying temperature, and caliber size. The models predicted that maximum k and Y0 -YEND, -2.62 ± 0.14 log10 CFU/g/day and 4.5 ± 0.1 log10 CFU/g, respectively, would be obtained fermenting at 30°C and drying at 20°C regardless of caliber. Drying at 14°C allowed Listeria growth until a water activity (aw) of 0.92 was reached. Therefore, if initial Listeria contamination is high (3 log10 CFU/g), drying at low temperatures will compromise product safety.

Effect of beef long-storage under different temperatures and vacuum-packaging conditions on meat quality, oxidation processes and microbial growth.

BACKGROUND: The global beef market demands the meat industry to ensure product quality and safety in markets that are often very distant. The present study aimed to evaluate the effects of chilled (CH, 120 d) and chilled-then-frozen (CHF, 28 d + 92 d) storage conditions of beef vacuum packaged (VP) and vacuum packaged with antimicrobial (VPAM) on meat quality, oxidative status and microbial loads. Treatments resulted from the combination of storage condition and packaging type: VP + CH, VP + CHF, VPAM + CH and VPAM + CHF. RESULTS: Warner-Bratzler shear force values decreased in all treatments after 28 d of chilling. Except for VP + CH, L* values (lightness) of meat color did not differ in each treatment as the storage time increased. Meat from VP + CH had greater a* values than CHF treatments on day 120 of storage. A consumer panel did not detect differences in tenderness, flavor and overall liking between VP and VPAM beef, but they preferred CHF steaks rather than CH beef. TBARS values did not differ between VP and VPAM and between CH and CHF at any time during the storage period. At the end of storage time, all treatments except VP + CHF presented a greater concentration of thiols than at 48 h post-mortem. On day 120 of storage, VP + CH had greater catalase enzyme activity than CHF treatments while VP + CH and VP + CHF showed a greater superoxide dismutase activity than VPAM + CHF. Storage condition (CH or CHF) had a greater impact on microbial counts than the type of packaging. CONCLUSION: Freezing meat after an ageing period represents a suitable strategy to extend beef storage life without a detrimental impact on its quality. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Dermatan Sulfate/Chitosan Nanoparticles Loaded with an Anti-Inflammatory Peptide Increase the Response of Human Colorectal Cancer Cells to 5-Fluorouracil.

The gold standard drug for colorectal cancer (CRC) treatment, 5-Fluorouracil (5-FU), induces pharmacological tolerance in long-term management. The transcriptional factor nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB) plays a key role in 5-FU resistance. The aim of this work is to study the capability of polyelectrolytes complex nanoparticles of dermatan sulfate (DS) and chitosan (CS), loaded with the anti-inflammatory tripeptide IRW, to sensitize colorectal cancer cells to 5-FU. Fluorescence and flow cytometry studies confirmed the recognition by the nanoformulation, of the cluster of differentiation 44 (CD44) receptor, involved in the initiation and progression of colorectal tumors. Dynamic light scattering (DLS) and flow cytometry reinforced the importance of DS and CD44 receptor in the interaction, as the addition of DS or anti-CD44 antibody blocked the binding. Moreover, the nanoformulation also interacts with 3D colon cancer cultures, namely colonospheres, enriched in cancer stem cells (CSC), subpopulation responsible for drug resistance and metastasis. To evaluate the consequences of this interaction, the subcellular distribution of the transcriptional factor NFκB, is determined by immunofluorescence analysis. Internalization and the intracellular release of IRW inhibited nuclear translocation of NFκB and increased cellular sensitivity to 5-FU. Altogether, the nanoformulation could provide a selective delivery platform for IRW distribution to colorectal tumors, being an innovative strategy toward overcoming 5-FU resistance in CRC therapy.

Wastewater surveillance of SARS-CoV-2 genomic populations on a country-wide scale through targeted sequencing.

SARS-CoV-2 surveillance of viral populations in wastewater samples is recognized as a useful tool for monitoring epidemic waves and boosting health preparedness. Next generation sequencing of viral RNA isolated from wastewater is a convenient and cost-effective strategy to understand the molecular epidemiology of SARS-CoV-2 and provide insights on the population dynamics of viral variants at the community level. However, in low- and middle-income countries, isolated groups have performed wastewater monitoring and data has not been extensively shared in the scientific community. Here we report the results of monitoring the co-circulation and abundance of variants of concern (VOCs) of SARS-CoV-2 in Uruguay, a small country in Latin America, between November 2020-July 2021 using wastewater surveillance. RNA isolated from wastewater was characterized by targeted sequencing of the Receptor Binding Domain region within the spike gene. Two computational approaches were used to track the viral variants. The results of the wastewater analysis showed the transition in the overall predominance of viral variants in wastewater from No-VOCs to successive VOCs, in agreement with clinical surveillance from sequencing of nasal swabs. The mutations K417T, E484K and N501Y, that characterize the Gamma VOC, were detected as early as December 2020, several weeks before the first clinical case was reported. Interestingly, a non-synonymous mutation described in the Delta VOC, L452R, was detected at a very low frequency since April 2021 when using a recently described sequence analysis tool (SAM Refiner). Wastewater NGS-based surveillance of SARS-CoV-2 is a reliable and complementary tool for monitoring the introduction and prevalence of VOCs at a community level allowing early public health decisions. This approach allows the tracking of symptomatic and asymptomatic individuals, who are generally under-reported in countries with limited clinical testing capacity. Our results suggests that wastewater-based epidemiology can contribute to improving public health responses in low- and middle-income countries.

Microbiological Changes during Long-Storage of Beef Meat under Different Temperature and Vacuum-Packaging Conditions.

We evaluated a combination of two temperatures and two packaging materials for long-term storage of vacuum-packaged (VP) beef striploins. Microbial populations and microbiome composition were monitored during refrigerated storage (120 days between 0-1.5 °C) and refrigerated-then-frozen storage (28 days between 0-1.5 °C then 92 days at -20 °C) under low-O2 permeability VP and high-O2 permeability VP with an antimicrobial (VPAM). Pseudomonas (PSE) and Enterobacteriaceae (EB) counts in VPAM samples were significantly higher (p < 0.05) than in VP samples at 28, 45, 90, and 120 days of storage. Microbiome data showed that bacteria of the genera Serratia and Brochothrix were more abundant in VPAM samples at 120 days, while lactic acid bacteria (LAB) dominated in VP samples. Frozen temperatures inhibited microbial growth and maintained a relatively stable microbiome. Refrigerated and frozen VPAM samples showed the greatest difference in the predicted metabolic functions at the end of storage driven by the microbiome composition, dominated by PSE and LAB, respectively. Although no signs of visible meat deterioration were observed in any sample, this study suggests that VP meat refrigerated and then frozen achieved better microbiological indicators at the end of the storage period.

Clinical experience with orphan drugs for rare metabolic diseases.

INTRODUCTION: Inborn errors of metabolism are a highly heterogeneous group of orphan diseases. Diet therapy and enzyme and coenzyme replacement are the most frequently used treatment. There are few patients and published studies about inborn errors of metabolism. The main objective of this study was to describe the effectiveness of orphan drugs in inborn errors of metabolism in paediatric patients. MATERIAL AND METHODS: Retrospective descriptive study of 24 months on patients diagnosed with inborn errors of metabolism during childhood and who attended the pharmacy clinic or Day-Care Unit of a 630-bed general hospital. RESULTS: The study included 15 patients with a median age of 17.8 years and were treated with nine different drugs: sapropterin, sodium phenylbutyrate, miglustat, velaglucerase, sebelipase, idursulfase, 5-hydroxytryptophan, succinate, and riboflavin. Seven different inborn errors of metabolism were observed: phenylketonuria, defects of the urea cycle, Gaucher, Nieman-Pick, Hunter's disease, along with acid lipase deficiency, and mitochondrial diseases. Orphan drugs used for the treatment of inborn errors of metabolism accounted for 1.3% of hospital drug costs. Some orphan drugs achieved asymptomatic patients, but others just produced a modest symptomatic improvement. Most patients showed good tolerance to the treatment. CONCLUSIONS: Orphan drugs used in inborn errors of metabolism had an easy to manage toxicity profile, with many disparities in effectiveness. These drugs have a high economic impact. The cost-effectiveness ratio for orphan drugs is a controversial issue due to their high cost and the inconclusive clinical evidence.

Effect of UV-C Irradiation and Lactic Acid Application on the Inactivation of Listeria monocytogenes and Lactic Acid Bacteria in Vacuum-Packaged Beef.

The objective of this study was to test the effect of the combined application of lactic acid (0-5%) (LA) and UV-C light (0-330 mJ/cm2) to reduce Listeria monocytogenes and lactic acid bacteria (LAB) on beef without major meat color (L *, a *, b *) change and its impact over time. A two-factor central composite design with five central points and response surface methodology (RSM) were used to optimize LA concentration and UV-C dose using 21 meat pieces (10 g) inoculated with L. monocytogenes (LM100A1). The optimal conditions were analyzed over 8 weeks. A quadratic model was obtained that predicted the L. monocytogenes log reduction in vacuum-packed beef treated with LA and UV-C. The maximum log reduction for L. monocytogenes (1.55 ± 0.41 log CFU/g) and LAB (1.55 ± 1.15 log CFU/g) with minimal impact on meat color was achieved with 2.6% LA and 330 mJ/cm2 UV-C. These conditions impaired L. monocytogenes growth and delayed LAB growth by 2 weeks in vacuum-packed meat samples throughout 8 weeks at 4 °C. This strategy might contribute to improving the safety and shelf life of vacuum-packed beef with a low impact on meat color.

[Clinical experience with orphan drugs for rare metabolic diseases]. / Experiencia clínica con medicamentos huérfanos para enfermedades raras metabólicas.

INTRODUCTION: Inborn errors of metabolism are a highly heterogeneous group of orphan diseases. Diet therapy and enzyme and coenzyme replacement are the most frequently used treatment. There are few patients and published studies about inborn errors of metabolism. The main objective of this study was to describe the effectiveness of orphan drugs in inborn errors of metabolism in paediatric patients. MATERIAL AND METHODS: Retrospective descriptive study of 24 months on patients diagnosed with inborn errors of metabolism during childhood and who attended the pharmacy clinic or Day-Care Unit of a 630-bed general hospital. RESULTS: The study included 15 patients with a median age of 17.8 years and were treated with nine different drugs: sapropterin, sodium phenylbutyrate, miglustat, velaglucerase, sebelipase, idursulfase, 5-hydroxytryptophan, succinate, and riboflavin. Nine different inborn errors of metabolism were observed: phenylketonuria, defects of the urea cycle, Gaucher, Nieman-Pick, Hunter's disease, along with acid lipase deficiency, and mitochondrial diseases. Orphan drugs used for the treatment of inborn errors of metabolism accounted for 1.3% of hospital drug costs. Some orphan drugs achieved asymptomatic patients, but others just produced a modest symptomatic improvement. Most patients showed good tolerance to the treatment. CONCLUSIONS: Orphan drugs used in inborn errors of metabolism had an easy to manage toxicity profile, with many disparities in effectiveness. These drugs have a high economic impact. The cost-effectiveness ratio for orphan drugs is a controversial issue due to their high cost and the inconclusive clinical evidence.

Seasonal Distribution and Movement of the Invasive Pest Delottococcus aberiae (Hemiptera: Pseudococcidae) Within Citrus Tree: Implications for Its Integrated Management.

Delottococcus aberiae (De Lotto) (Hemiptera: Pseudococcidae) is the most recent species of mealybug introduced to Spain that is affecting citrus. The feeding behavior of D. aberiae causes severe direct damage to citrus fruits, distorting their shape and/or causing reduction in size. There is no information available regarding its distribution within the citrus trees. The main objective of this study was to describe the seasonal distribution of D. aberiae within citrus trees and its migration patterns on the plants. Ten citrus orchards from eastern Spain were periodically sampled during 3 yr. In each orchard, the mealybug was sampled in different infested strata (canopy, trunk, and soil) and canopy structures (flower, fruit, leaf, and twig). Results showed that, within the sampled strata, D. aberiae was mostly in the canopy. Within the canopy, the feeding location of D. aberiae changed throughout the year. D. aberiae overwintered in the twigs and moved to the flowers and fruits in spring. Once there, its populations started to increase exponentially until August. From February to September, 5-30% of the mealybugs migrated to the trunk and soil. These results will facilitate an early detection of the pest in the areas where it is spreading and improve sampling protocols and pesticide applications.

Increased Na⁺/Ca²âº exchanger expression/activity constitutes a point of inflection in the progression to heart failure of hypertensive rats.

UNLABELLED: Spontaneously hypertensive rat (SHR) constitutes a genetic model widely used to study the natural evolution of hypertensive heart disease. Ca²âº-handling alterations are known to occur in SHR. However, the putative modifications of Ca²âº-handling proteins during the progression to heart failure (HF) are not well established. Moreover, the role of apoptosis in SHR is controversial. We investigated intracellular Ca²âº, Ca²âº-handling proteins and apoptosis in SHR vs. control Wistar rats (W) from 3 to 15 months (mo). Changes associated with the transition to HF (i.e. lung edema and decrease in midwall fractional shortening), occurred at 15 mo in 38% of SHR (SHRF). In SHRF, twitch and caffeine-induced Ca²âº transients, significantly decreased relative to 6/9 mo and 15 mo without HF signs. This decrease occurred in association with a decrease in the time constant of caffeine-Ca²âº transient decay and an increase in Na⁺/Ca²âº exchanger (NCX) abundance (p<0.05) with no changes in SERCA2a expression/activity. An increased Ca²âº-calmodulin-kinase II activity, associated with an enhancement of apoptosis (TUNEL and Bax/Bcl2) was observed in SHR relative to W from 3 to 15 mo. CONCLUSIONS: 1. Apoptosis is an early and persistent event that may contribute to hypertrophic remodeling but would not participate in the contractile impairment of SHRF. 2. The increase in NCX expression/activity, associated with an increase in Ca²âº efflux from the cell, constitutes a primary alteration of Ca²âº-handling proteins in the evolution to HF. 3. No changes in SERCA2a expression/activity are observed when HF signs become evident.

Componentes descriptivos y explicativos de la accidentalidad vial en Colombia: incidencia del factor humano / Descriptive and explanatory components of road accident rate in Colombia: Influence of the human factor / Componentes descritivos e explicativos da acidentalidade rodoviária na Colômbia: incidência do fator humano

Problema: Se analizó la incidencia del factor humano en la accidentalidad vial en el territorio colombiano, y se identificó evidencia empírica para la formulación de política pública del tránsito. Metodología: El diseño es descriptivo-correlacional. Instrumentos: cuestionario de comportamiento para conductores y motociclistas (D.B.Q.) y encuesta tipo Likert. Muestra no probabilística intencional: 16.322 personas (8.631 conductores de automotores, 5.133 motociclistas y 2.558 peatones, pasajeros y acompañantes). Resultados: a) conductores con nivel educativo superior inciden en menos accidentes; b) peatones con menor nivel educativo inciden en mayores conductas riesgosas; c) las mujeres tienen actitud positiva y perciben eficacia de campañas en prevencion; d) estilos de conducción iracundo, ansioso, riesgoso y de alta velocidad cometen más infracciones y accidentes; e) desobedecer señales de tránsito, la principal causa de accidentalidad; f) agresividad, hostilidad y estrés en el tránsito son factores que aumentan la probabilidad de accidente; g) campañas de prevención no están siendo captadas por la población más afectada. Conclusiones y recomendaciones: Los factores de accidentalidad vial guardan correspondencia con el Triángulo de Seguridad Vial establecido por la Organización de las Naciones Unidas –factor humano preponderante en la accidentalidad–. Lineamientos de política pública fundamentados en procesos educativos y corporativos tendientes a disminuir la accidentalidad por lo general no se concibe aplicada a la seguridad pública. Por ello, se presenta la inteligencia criminal como una disciplina que se ocupa de anticipar los riesgos criminales contra la seguridad pública. Esto permitiría disminuir la incertidumbre durante la toma de decisiones y calcular los daños contra la seguridad pública, que se pretende prevenir.

Problem: The influence of the human factor on accident rates in the Colombian territory was analyzed, and empirical evidence was identified for the formulation of public traffic policy. Methodology: The design is descriptive-correlational. Instruments: A behavior questionnaire (the D.B.Q.) for car drivers and motorcyclists, and a Likert-type survey. An intentional non-probabilistic sample: 16,322 individuals (8,631 vehicle drivers, 5,133 motorcyclists and 2,558 pedestrians, passengers, and accompanying persons). Results: a) drivers with a higher educational level have a lower impact on the accident rate; b) pedestrians with a lower educational level exhibit more risky behaviors: c) women have a poenfusitive attitude and perceive the effectiveness of prevention campaigns; d) angry, anxious, risky and high-speed driving result in more traffic offenses and accidents; e) disobeying traffic signs is the main cause of accidents; f) aggressiveness, hostility and stress are factors tending to increase the likelihood of traffic accidents; g) prevention campaigns are not being heard by the more affected population. Conclusions and recommendations: The accident rate factors correspond to the Road Safety Triangle established by the UNO –human factor prominent in accident levels–. Public policy guidelines based on educational and corporative processes leading to reduce the accident rate.

Problema: A incidência do fator humano na acidentalidade rodoviária no território colombiano foi analisada e a evidência empírica para a formulação da política pública do tráfego foi identificada. Metodologia: O projeto é descritivo-correlacional. Instrumentos: questionário do comportamento para os motoristas e os motoqueiros (D.B.Q.) e pesquisa tipo Likert. Amostra intencional não-probabilística: 16.322 pessoas (8.631 condutores de automotores, 5.133 motoqueiros e 2.558 pedestres, passageiros e companheiros). Resultados: a) os condutores com nível educativo superior incidem em menos acidentes; b) os pedestres com nível educativo menor incidem em maiores condutas de risco; c) as mulheres têm uma atitude positiva e percebem a eficácia das campanhas na prevenção; d) os estilos de dirigir enfurecido, ansioso, riscoso e da alta velocidade contribuem a cometer mais infrações e acidentes; e) para desobedecer sinalizações rodoviárias, a causa principal da acidentalidade; f) a agressividade, a hostilidade e o stress no tráfego são os fatores que aumentam a probabilidade do acidente; g) as campanhas da prevenção não são captadas pela população afetada. Conclusões e recomendações: Os fatores da acidentalidade rodoviária correspondem ao Triângulo da Segurança Rodoviária estabelecido pelas Nações Unidas – fator humano preponderante na acidentalidade–. Diretrizes da política pública baseadas em processos educativos e corporativos tendentes a diminuir a acidentalidade.

Microgastrinae (Hymenoptera: Braconidae) in the Forest State of Artikutza (Navarra: Spain): Diversity and Community Structure.

Microgastrinae is one of the largest subfamilies of the Braconidae with about 2,000 described species worldwide. These wasps are of enormous ecological interest due to their role in controlling the caterpillar populations. This study analyses diversity and community structure within the Microgastrinae in the Artikutza Forest, located in the Peñas de Aia Natural Park, western Pyrenees, Spain. The specimens were collected in two different habitats: mixed forest and beech forest. A total of 524 specimens, belonging to nine separate genera and 27 species were captured. Alpha, beta and gamma diversity were analyzed. Additionally, the relationship between Microgastrinae phenology and climatic conditions were studied.

The signalling pathway of CaMKII-mediated apoptosis and necrosis in the ischemia/reperfusion injury.

Ca(2+)-calmodulin-dependent protein kinase II (CaMKII) plays an important role mediating apoptosis/necrosis during ischemia-reperfusion (IR). We explored the mechanisms of this deleterious effect. Langendorff perfused rat and transgenic mice hearts with CaMKII inhibition targeted to sarcoplasmic reticulum (SR-AIP) were subjected to global IR. The onset of reperfusion increased the phosphorylation of Thr(17) site of phospholamban, without changes in total protein, consistent with an increase in CaMKII activity. Instead, there was a proportional decrease in the phosphorylation of Ser2815 site of ryanodine receptors (RyR2) and the amount of RyR2 at the onset of reperfusion, i.e. the ratio Ser2815/RyR2 did not change. Inhibition of the reverse Na(+)/Ca(2+)exchanger (NCX) mode (KBR7943) diminished phospholamban phosphorylation, reduced apoptosis/necrosis and enhanced mechanical recovery. CaMKII-inhibition (KN-93), significantly decreased phospholamban phosphorylation, infarct area, lactate dehydrogenase release (LDH) (necrosis), TUNEL positive nuclei, caspase-3 activity, Bax/Bcl-2 ratio and Ca(2+)-induced mitochondrial swelling (apoptosis), and increased contractile recovery when compared with non-treated IR hearts or IR hearts pretreated with the inactive analog, KN-92. Blocking SR Ca(2+) loading and release (thapsigargin/dantrolene), mitochondrial Ca(2+) uniporter (ruthenium red/RU360), or mitochondrial permeability transition pore (cyclosporine A), significantly decreased infarct size, LDH release and apoptosis. SR-AIP hearts failed to show an increase in the phosphorylation of Thr(17) of phospholamban at the onset of reflow and exhibited a significant decrease in infarct size, apoptosis and necrosis respect to controls. The results reveal an apoptotic-necrotic pathway mediated by CaMKII-dependent phosphorylations at the SR, which involves the reverse NCX mode and the mitochondria as trigger and end effectors, respectively, of the cascade.