High prevalence of long-term olfactory, gustatory, and chemesthesis dysfunction in post-COVID-19 patients: a matched case-control study with one-year follow-up using a comprehensive psychophysical evaluation.

BACKGROUND: Using an age and gender matched-pair case-control study, we aimed to estimate the long-term prevalence of psychophysical olfactory, gustatory , and chemesthesis impairment at least one year after SARS-CoV-2 infection considering the background of chemosensory dysfunction in non-COVID-19 population. METHODOLOGY: This case-controlled study included 100 patients who were home-isolated for mildly symptomatic COVID-19 between March and April 2020. One control regularly tested for SARS-CoV-2 infection and always tested negative was matched to each case according to gender and age. Chemosensory function was investigated by a comprehensive psychophysical evaluation including ortho- and retronasal olfaction and an extensive assessment of gustatory function. Differences in chemosensory parameters were evaluated through either Fisher’s exact test or Kruskal-Wallis test. RESULTS: The psychophysical assessment of chemosensory function took place after a median of 401 days from the first SARS-CoV-2 positive swab. The evaluation of orthonasal smell identified 46% and 10% of cases and controls, respectively, having olfactory dysfunction, with 7% of COVID-19 cases being functionally anosmic. Testing of gustatory function revealed a 27% of cases versus 10% of controls showing a gustatory impairment. Nasal trigeminal sensitivity was significantly lower in cases compared to controls. Persistent chemosensory impairment was associated with emotional distress and depression. CONCLUSION: More than one year after the onset of COVID-19, cases exhibited an excess of olfactory, gustatory , and chemesthesis disturbances compared to matched-pair controls with these symptoms being associated to emotional distress and depression.

Lactic acid bacteria biofilms and their ability to mitigate Escherichia coli O157:H7 surface colonization.

Lactic acid bacteria (LAB) exert antagonistic activities against diverse microorganisms, including pathogens. In this work, we aimed to investigate the ability of LAB strains isolated from food to produce biofilms and to inhibit growth and surface colonization of Enterohaemorrhagic Escherichia coli (EHEC) O157:H7 at 10°C. The ability of 100 isolated LAB to inhibit EHEC O157:H7 NCTC12900 growth was evaluated in agar diffusion assays. Thirty-seven LAB strains showed strong growth inhibitory effect on EHEC. The highest inhibitory activities corresponded to LAB strains belonging to Lactiplantibacillus plantarum, Pediococcus acidilactici and Pediococcus pentosaceus species. Eighteen out of the 37 strains that showed growth inhibitory effects on EHEC also had the ability to form biofilms on polystyrene surfaces at 10°C and 30°C. Pre-established biofilms on polystyrene of four of these LAB strains were able to reduce significantly surface colonization by EHEC at low temperature (10°C). Among these four strains, Lact. plantarum CRL 1075 not only inhibited EHEC but also was able to grow in the presence of the enteric pathogen. Therefore, this strain proved to be a good candidate for further technological studies oriented to its application in food-processing environments to mitigate undesirable surface contaminations of E. coli.

Pleiotrophin, the angiogenic and mitogenic growth factor: levels in serum and synovial fluid in rheumatoid arthritis and osteoarthritis : And correlation with clinical, laboratory and radiological indices.

BACKGROUND: Few studies have reported a possible involvement of pleiotrophin (PTN) in the pathophysiology of osteoarthritis (OA) and very little is known about its role in rheumatoid arthritis (RA). This study is to measure PTN in the sera and synovial fluids in RA and OA and to assess its relation to activity, functional class and radiological staging. SUBJECTS AND METHODS: Serum and synovial fluid samples were collected from 35 RA patients and 40 knee OA patients and serum samples were withdrawn from 20 healthy controls. Demographic, clinical and serological data were prospectively assessed. Functional and radiographic grades were also assessed. Serum and synovial fluid PTN levels were measured using enzyme-linked immunosorbent assay (ELISA). RESULTS: There was no statistical significant differences (p > 0.05) on comparing the mean PTN level in sera of RA, OA patients and healthy controls. However the mean synovial fluid level of PTN in both patient groups was significantly higher than mean serum level (p < 0.001). Significant correlations between the serum PTN level and both morning stiffness duration (p = 0.008) and mHAQ score (p = 0.039) were only observed in RA patients. CONCLUSION: Our results point to a possible important role of PTN in RA and OA. We firstly report a serological pattern of PTN in the sera and synovial fluids of RA patients. However its implementation as a disease marker or a potential target therapy in both diseases awaits larger studies and further investigations.

Subclinical atherosclerosis in systemic lupus erythematosus patients and its relationship to disease activity and damage indices.

AIM OF THE WORK: To detect the incidence of premature atherosclerosis in systemic lupus erythematosus (SLE) patients and to study its association with disease activity and damage indices. PATIENTS AND METHODS: This study involved 50 adult female SLE patients with mean age 26.24 ± 8.63 years and mean disease duration 3.44 ± 4.01 years. The control group comprised 25 healthy adult females. All patients were subjected to a detailed clinical examination and laboratory investigations, and full case history was recorded. Assessment of disease activity was performed according to the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) and disease damage was assessed using the Systemic Lupus International Collaborating Clinics (SLICC) score. B mode ultrasound was used to measure the intima-media wall thickness (IMT) and detect the presence of carotid plaques. RESULTS: In 15 patients (30 %), positive ultrasonographic findings represented by a significant increase in IMT (> 0.9 mm) could be shown; plaques were found in 3 of these patients (6 %). A significant difference was found between SLE patients and controls in terms of IMT (P < 0.0001). On subgrouping the SLE patients according to their IMT, there was a significant difference between those with thickened and normal IMT in terms of SLEDAI (P < 0.0001) and SLICC (p = 0.035) scores. CONCLUSION: Subclinical atherosclerosis is frequent in SLE patients. Increased disease activity and damage are associated with the occurrence of premature atherosclerosis.

Resistin in inflammatory and degenerative rheumatologic diseases. Relationship between resistin and rheumatoid arthritis disease progression.

AIMS OF THE STUDY: To assess and compare resistin levels in the serum and synovial fluid of patients with rheumatoid arthritis (RA; an inflammatory rheumatologic disease) and osteoarthritis (OA; a degenerative rheumatologic disease) and to study the relationship between resistin levels and prognostic factors of RA disease progression. PATIENTS AND METHODS: This study included a total of 50 patients: 25 with RA and 25 with OA. Full case history was documented for all patients and all underwent a thorough clinical examination and laboratory testing. Body mass index (BMI) values were also calculated. Radiographs were made of OA patients' knees and RA patients' hands. Disease Activity Score 28 (DAS28) was calculated for RA patients. Serum and synovial fluid samples were obtained from the effused knees of all patients and tested for resistin level. RESULTS: Serum resistin levels were higher in RA patients than in those with OA (p < 0.01). Synovial fluid resistin levels were also higher in RA than OA patients (p < 0.001). While serum resistin levels correlated with Larsen score and total leukocyte count (TLC), synovial fluid resistin levels correlated with rheumatoid factor (RF) and anti-citrullinated protein antibody (ACPA) levels in addition to Larsen score and TLC. CONCLUSION: Resistin levels were found to be higher in the serum and synovial fluid of RA patients than in those with OA. This may suggest a role for resistin in inflammatory rheumatologic diseases. The observed statistically significant correlation between synovial fluid resistin levels and RF, ACPA and Larsen score may suggest that high synovial fluid resistin levels can be considered a poor prognostic factor for RA progression. However, further studies employing a larger cohort of patients are needed to confirm the relevance of resistin as a prognostic marker in RA patients.

The effect of EIF dynamics on the cryopreservation process of a size distributed cell population.

Typical mathematical modeling of cryopreservation of cell suspensions assumes a thermodynamic equilibrium between the ice and liquid water in the extracellular solution. This work investigates the validity of this assumption by introducing a population balance approach for dynamic extracellular ice formation (EIF) in the absence of any cryo-protectant agent (CPA). The population balance model reflects nucleation and diffusion-limited growth in the suspending solution whose driving forces are evaluated in the relevant phase diagram. This population balance description of the extracellular compartment has been coupled to a model recently proposed in the literature [Fadda et al., AIChE Journal, 56, 2173-2185, (2010)], which is capable of quantitatively describing and predicting internal ice formation (IIF) inside the cells. The cells are characterized by a size distribution (i.e. through another population balance), thus overcoming the classic view of a population of identically sized cells. From the comparison of the system behavior in terms of the dynamics of the cell size distribution it can be concluded that the assumption of a thermodynamic equilibrium in the extracellular compartment is not always justified. Depending on the cooling rate, the dynamics of EIF needs to be considered.

In vitro ovine articular chondrocyte proliferation: experiments and modelling.

This study focuses on analysis of in vitro cultures of chondrocytes from ovine articular cartilage. Isolated cells were seeded in Petri dishes, then expanded to confluence and phenotypically characterized by flow cytometry. The sigmoidal temporal profile of total counts was obtained by classic haemocytometry and corresponding cell size distributions were measured electronically using a Coulter Counter. A mathematical model recently proposed (1) was adopted for quantitative interpretation of these experimental data. The model is based on a 1-D (that is, mass-structured), single-staged population balance approach capable of taking into account contact inhibition at confluence. The model's parameters were determined by fitting measured total cell counts and size distributions. Model reliability was verified by predicting cell proliferation counts and corresponding size distributions at culture times longer than those used when tuning the model's parameters. It was found that adoption of cell mass as the intrinsic characteristic of a growing chondrocyte population enables sigmoidal temporal profiles of total counts in the Petri dish, as well as cell size distributions at 'balanced growth', to be adequately predicted.

Experimental analysis and modelling of in vitro proliferation of mesenchymal stem cells.

OBJECTIVES: Stem cell therapies based on differentiation of adult or embryonic stem cells into specialized ones appear to be effective for treating several human diseases. This work addresses the mathematical simulation of proliferation kinetics of stem cells. MATERIALS AND METHODS: Sheep bone marrow mesenchymal stem cells (phenotype characterized by flow cytometry analysis) seeded at different initial concentrations in Petri dishes were expanded to confluence. Sigmoid temporal profiles of total counts obtained through classic haemocytometry were quantitatively interpreted by both a phenomenological logistic equation and a novel model based on a one-dimensional, single-staged population balance approach capable of taking into account contact inhibition at confluence. The models' parameters were determined by comparison with experimental data on population expansion starting from single seeding concentration. Reliability of the models was tested by predicting cell proliferation carried out starting from different seeding concentrations. RESULTS AND DISCUSSION: It was found that the proposed population balance modelling approach was successful in predicting the experimental data over the whole range of initial cell numbers investigated, while prediction capability of phenomenological logistic equation was more limited.

A review of bacteriocinogenic lactic acid bacteria used as bioprotective cultures in fresh meat produced in Argentina.

Several lactic acid bacteria (LAB) associated with meat products are important natural bacteriocin producers. Bacteriocins are proteinaceous antagonistic substances that are important in the control of spoilage and pathogenic microorganisms. The use of LAB as bioprotective cultures to extend the shelf life of fresh meat can improve microbial stability and safety in commercial meat preservation. Lactobacillus curvatus CRL705 used as a protective culture in fresh beef is effective in inhibiting Listeria innocua and Brochothrix thermosphacta as well as the indigenous contaminant LAB, retaining its inhibitory effect at low temperatures and having a negligible effect on meat pH. In addition to the hurdle represented by low temperature and vacuum-packaging, the use of live cells of Lb. curvatus CRL705 seems more feasible from an economic point of view - and without legal restrictions - compared to the addition of purified bacteriocins. A description of meat-borne bacteriocins and their application in meat to extend shelf life is discussed.

Lactobacillus role during conditioning of refrigerated and vacuum-packaged Argentinean meat.

The role of Lactobacillus strains with bioprotective and technological potential on raw beef during 15days of storage under vacuum at 7°C was investigated. The assayed strains were able to grow on the meat, Lactobacillus curvatus CRL705 and Lactobacillus sakei 23K showing the highest competitiveness. A net increase of amino acids was determined in inoculated samples when compared to the control, this being maximal for Lactobacillus plantarum CRL681. Although an important endogenous activity of meat sarcoplasmic proteins was observed, the disappearance of protein bands and the generation of a new one were detected as a consequence of Lactobacillus growth. A synergistic effect of Lactobacillus in combination with the muscle proteolytic enzyme complex can be suggested. From the studied strains, the bacteriocin producer L. curvatus CRL705 may be considered as a good candidate to contribute to meat ageing by means of small peptides and free amino acids generation while improving shelf life.

Conceptual basis and methodology of the SOPHIA study.

To clarify the role of gene polymorphisms on the effect of losartan and losartan plus hydrochlorothiazide on blood pressure (primary end point) and on cardiac, vascular and metabolic phenotypes (secondary end point) after 4, 8, 12, 16 and 48 weeks treatment, an Italian collaborative study - The Study of the Pharmacogenomics in Italian hypertensive patients treated with the Angiotensin receptor blocker losartan (SOPHIA) - on never-treated essential hypertensives (n = 800) was planned. After an 8 week run-in, losartan 50 mg once daily will be given and doubled to 100 mg at week +4 if blood pressure is more than 140/90 mmHg. Hydroclorothiazide 25 mg once daily at week +8 and amlodipine 5 mg at week +16 will be added if blood pressure is more than 140/90 mmHg. Cardiac mass (echocardiography), carotid intima-media thickness, 24 h ambulatory blood pressure, homeostatic model assessment (HOMA) index, microalbuminuria, plasma renin activity and aldosterone, endogenous lithium clearance, brain natriuretic peptide and losartan metabolites will be evaluated. Genes of the renin-angiotensin-aldosterone system, salt sensitivity, the beta-adrenergic system and losartan metabolism will be studied (Illumina custom arrays). A whole-genome scan will also be performed in half of the study cohort (1M array, Illumina 500 GX beadstation).

Surface properties and behaviour on abiotic surfaces of Staphylococcus carnosus, a genetically homogeneous species.

This work aimed to characterize the surface properties of Staphylococcus carnosus and the influence of different media on their ability to adhere and grow on industrial supports. As their colonization could be dependant of the strain, the genetic diversity of the strains was studied. The diversity of 13 strains analysed by pulsed-field gel electrophoresis revealed that the S. carnosus strains formed a homogeneous genetic group. Their surface properties, characterized by studying their affinity to solvents, were hydrophilic with a strong negative surface charge. The S. carnosus strain CIT 833 hardly adhered to polytetrafluoroethylene (PTFE) and stainless steel chips. Tryptic soy broth (TSB) was the most favourable medium for growth on stainless steel support while TSB/NaCl was better for growth on PTFE. Scanning electron microscopy (sem) showed that this strain weakly colonized both supports and did not form cell aggregates. Indeed, the strain did not synthesize polysaccharides. These results showed that S. carnosus adhered on different abiotic surfaces which are used in food factories but was not able to accumulate on these surfaces. The inability of S. carnosus to form biofilm could explain why S. carnosus is rarely isolated in meat processing environment.

Decarboxylase activity involved in methyl ketone production by Staphylococcus carnosus 833, a strain used in sausage fermentation.

Staphylococcus carnosus strain 833, inoculated into sausage, increased the levels of methyl ketones which contributed to the cured aroma. These ketones were predicted to arise from incomplete beta-oxidation followed by a decarboxylation. To check this hypothesis, we measured the beta-decarboxylase activity in resting cells of S. carnosus grown in complex or in synthetic medium, using as substrate a beta-ketoacid, which can be an intermediate of the beta-oxidation pathway. This activity was present throughout the growth period. The enzyme appeared to be constitutive because no induction was observed. High aeration, a pH of 5 and the presence of nitrate promoted the production of methyl ketones.

Effect of curing conditions and Lactobacillus casei CRL705 on the hydrolysis of meat proteins.

AIMS: The effect of the common curing conditions used during the manufacture of dry fermented sausage on the proteolytic activity of Lactobacillus casei CRL705 against meat proteins was investigated. METHODS AND RESULTS: Hydrolysis of pork muscle sarcoplasmic and myofibrillar proteins was evaluated by SDS-PAGE and reverse phase-HPLC analysis. Ascorbic acid exerted a stimulatory effect on both sarcoplasmic and myofibrillar protein breakdown by Lact. casei CRL705 with the release of hydrophilic peptides and free amino acids, while NaCl and NaNO2 mainly stimulated myofibrillar degradation. CONCLUSION: Even when processing temperature (25 degrees C) did not positively affect bacterial protein hydrolysis, the presence of curing salts accounted for a remarkable increase in the non-volatile components that constitute taste-active compounds that strongly influence the final flavour of the product. SIGNIFICANCE AND IMPACT OF THE STUDY: To predict the suitability of Lact. casei CRL705 and its proteolytic enzymes as a starter culture for the dry processing of dry fermented sausages.

Soil decontamination at the Montevecchio-Levante mine site with experimental washing and leaching techniques.

The soils in the neighbourhood of the Rio Montevecchio-Sitzerri, a stream that flows in the valley below the tailings pond of the Montevecchio-Levante mineral processing plant (SW Sardinia, Italy) are severely contaminated by heavy metals, to the extent that traditional land uses are compromised. Consequently urgent measures are needed both to abate the pollution at source and rehabilitate the contaminated land. This paper is concerned with the problem of soil decontamination using washing and leaching techniques. Laboratory experiments have been conducted in mechanically agitated reactors, using citric acid and acetic acid solutions and brine of hydrochloric acid and calcium chloride. The influence of both reagent concentration and solid-to-liquid ratio has been assessed, and in the most significant cases, the attack kinetics has been determined. The tests showed the brine to be the most effective for removing metals from the soils. Based on the findings of the investigations, the possibility of decontamination by heap leaching has been simulated in the laboratory using the column technique.

Hydrolytic action of Lactobacillus casei CRL 705 on pork muscle sarcoplasmic and myofibrillar proteins.

Lactobacillus casei CRL 705 was screened, among other meat isolates, for its proteinase and aminopeptidase activities toward synthetic substrates and, according to that, selected for specific assays on muscle proteins. The hydrolytic effects of whole cells, cell free extracts (CFE), and the combination of both on muscle sarcoplasmic and myofibrillar protein extracts was evaluated by SDS-PAGE and reverse phase HPLC analyses. The proteinase activity of whole cells caused the degradation of a great number of sarcoplasmic protein bands. A partial hydrolysis was also associated with CFE that when combined with whole cells showed an important additional degradation. Peptide profiles from sarcoplasmic protein extracts were greatly modified regardless of the addition of whole cells or CFE, although their combination intensified these changes. The generation of free amino acids was remarkable when whole cells and CFE were incorporated together to sarcoplasmic protein extracts.

Characterization of muscle sarcoplasmic and myofibrillar protein hydrolysis caused by Lactobacillus plantarum.

Strains of Lactobacillus plantarum originally isolated from sausages were screened for proteinase and aminopeptidase activities toward synthetic substrates; on the basis of that screening, L. plantarum CRL 681 was selected for further assays on muscle proteins. The activities of whole cells, cell extracts (CE), and a combination of both on sarcoplasmic and myofibrillar protein extracts were determined by protein, peptide, and free-amino-acid analyses. Proteinase from whole cells initiated the hydrolysis of sarcoplasmic proteins. The addition of CE intensified the proteolysis. Whole cells generated hydrophilic peptides from both sarcoplasmic and myofibrillar proteins. Other peptides of a hydrophobic nature resulted from the combination of whole cells and CE. The action of both enzymatic sources on myofibrillar proteins caused maximal increases in lysine, arginine, and leucine, while the action of those on sarcoplasmic proteins mainly released alanine. In general, pronounced hydrolysis of muscle proteins required enzyme activities from whole cells in addition to those supplied by CE.

Hydrolysis of pork muscle sarcoplasmic proteins by lactobacillus curvatus and lactobacillus sake.

Lactobacillus curvatus CECT 904 and Lactobacillus sake CECT 4808 were selected on the basis of their proteolytic activities against synthetic substrates. Further, the effects of whole cells, cell extracts, and a combination of both enzymatic sources on muscle sarcoplasmic proteins were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and reverse-phase high-performance liquid chromatography analyses. Strains of both species displayed proteinase activities on five sarcoplasmic proteins. The inoculation of whole cells caused a degradation of peptides, whereas the addition of cell extracts resulted in the generation of both hydrophilic and hydrophobic peptides. This phenomenon was remarkably more pronounced when L. curvatus was involved. Whole cells also consumed a great amount of free amino acids, while the addition of intracellular enzymes contributed to their generation. L. sake accounted for a greater release of free amino acids. In general, cell viability and also proteolytic events were promoted when cell suspensions were provided with cell extracts as an extra source of enzymes.

Hydrolysis of muscle myofibrillar proteins by Lactobacillus curvatus and Lactobacillus sake.

Proteolytic enzyme activities of whole cells and cell free extracts (CFE) of Lactobacillus curvatus CECT 904 and Lactobacillus sake CECT 4808 were characterised using synthetic chromogenic compounds and myofibrillar proteins as substrates. The hydrolytic action was monitored by SDS-PAGE and reverse phase-HPLC analyses. The CFE of L. sake partially contributed, together with muscle enzymes, to the initial hydrolysis of myofibrillar proteins. Whole-cells of both L. curvatus and L. sake generated peptides considered important for cured-meat taste. The peptide mapping, resulting from the action on the substrates assayed, revealed a profile of extra and intracellular enzymes. Both strains expressed strong amino acid metabolism.

Proteolytic activity of Lactobacillus strains isolated from dryfermented sausages on muscle sarcoplasmic proteins.

The proteolytic activity of seven strains of Lactobacillus from two species isolated from dry cured sausages was assayed using a soluble muscle extract as a source of proteins, at a temperature of 30 °C. The results indicated that the strains of Lactobacillus plantarum tested had the more active proteolytic system, showing the highest amino acid release in the medium after 72 hr of incubation (L. plantarum CRL 681) when the microorganism was in the stationary phase of growth. The strains of L. casei showed a continued hydrolytic activity with a lower amino acids concentration along the studied period. The SDS-PAGE profiles showed that the major changes in sarcoplasmic proteins were produced in the 13 kDa and 36-40 kDa molecular weights region.