A neurotherapeutic approach with Lacticaseibacillus rhamnosus E9 on gut microbiota and intestinal barrier in MPTP-induced mouse model of Parkinson's disease.

The gut microbiota plays a crucial role in neural development and progression of neural disorders like Parkinson's disease (PD). Probiotics have been suggested to impact neurodegenerative diseases via gut-brain axis. This study aims to investigate the therapeutic potential of Lacticaseibacillus rhamnosus E9, a high exopolysaccharide producer, on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)-induced mouse model of PD. C57BL/6 mice subjected to MPTP were fed L. rhamnosus E9 for fifteen days and sacrificed after the last administration. Motor functions were determined by open-field, catalepsy, and wire-hanging tests. The ileum and the brain tissues were collected for ELISA, qPCR, and immunohistochemistry analyses. The cecum content was obtained for microbiota analysis. E9 supplementation alleviated MPTP-induced motor dysfunctions accompanied by decreased levels of striatal TH and dopamine. E9 also reduced the level of ROS in the striatum and decreased the DAT expression while increasing the DR1. Furthermore, E9 improved intestinal integrity by enhancing ZO-1 and Occludin levels and reversed the dysbiosis of the gut microbiota induced by MPTP. In conclusion, E9 supplementation improved the MPTP-induced motor deficits and neural damage as well as intestinal barrier by modulating the gut microbiota in PD mice. These findings suggest that E9 supplementation holds therapeutic potential in managing PD through the gut-brain axis.

Comparison of Self-Efficacy and Problem-Solving Skills Between Women with Asthma and Healthy Controls: A Cross-Sectional Study.

OBJECTIVE: Problem-solving skills and self-efficacy are among the topics that are frequently investigated in people with various chronic conditions. However, there are limited studies on asthma patients. Our study aims to compare self-efficacy and problem-solving skills in asthma patients and healthy controls. MATERIAL AND METHODS: We included 23 women with asthma [age: 39 (34-56) years] and 23 healthy controls [age: 42 (30-55) years] in the study. Participants' sociodemographic and disease-related characteristics, Asthma Control Test, and the Modified Medical Research Council Dyspnea Scale scores were recorded. We examined problem-solving skills with the Problem-Solving Inventory and self-efficacy with General Self-Efficacy Scale. We compared groups with the chi-square test, Mann-Whitney U-test, and Independent Sample t-test. RESULTS: Age, body mass index, educational status, marital status, and occupational status were similar among the groups (P > .05). However, smoking was significantly higher in healthy controls (P < .05). It was found that women with asthma had worse self-efficacy and problem-solving skills compared to healthy controls (P < .05). CONCLUSION: These results showed that there may be problems in socio-cognitive skills associated with the disease. Our study focused on the possibility that female asthma patients may have low levels of self-efficacy and problem-solving skills. Therefore, health professionals designing the rehabilitation program should take these skills into account while conducting the assessment as they may be useful in developing an efficient rehabilitation program.

Gut Microbial Alteration in MPTP Mouse Model of Parkinson Disease is Administration Regimen Dependent.

Parkinson Disease (PD) is one of the most common neurodegenerative disorders characterized by loss of dopaminergic neurons involved in motor functions. Growing evidence indicates that gut microbiota communicates with the brain known as the gut-brain axis (GBA). Mitochondrial toxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is commonly used in animal studies to investigate the GBA in PD. Various MPTP administration regimens are performed in PD mouse models involving one to multiple injections in 1 day or one injection per day for several days. The aim of this study is to investigate if the impact of MPTP on gut microbiota differs depending on the administration regimen. C57BL/6 mice were treated with acute or subchronic regimens of MPTP. Motor functions were assessed by open-field, catalepsy, and wire hanging tests. The cecum and the brain samples were obtained for microbiota and gene expression analyses, respectively. MPTP administration regimens differed in their ability to alter the gut microbiota. Firmicutes and Bacteroidota were both increased in subchronic mice while did not change and decreased, respectively, in acute mice. Verrucomicrobiota was elevated in acute MPTP mice but dropped in subchronic MPTP mice. Muribaculaceae was the predominant genus in all groups but acute mice. In acute mice, Akkermansia was increased and Colidextribacter was decreased; however, they showed an opposite trend in subchronic mice. These data suggest that MPTP mouse model cause a gut microbiota dysbiosis in an administration regimen dependent manner, and it is important to take consideration of mouse model to investigate the GBA in neurodegenerative diseases including PD.

Shelf-life studies of putative probiotic Lacticaseibacillus casei strains in milk and model yogurt.

Lacticaseibacillus casei are commonly utilized as probiotic in a wide-range of fermented and unfermented dairy products. The stability of probiotics in fermented dairy products during shelf-life is of concern due to low pH and high level of organic acids. The objective of this study is to evaluate L. casei for their ability to survive in a model yogurt and fluid milk; additionally, their impact on the pH, organic acids, and sensory attributes of these products was examined. The strain-to-strain differences in cell densities in yogurt and milk inoculated at a therapeutic level at the end of shelf-life were 1.2 and 1.4 log CFU/mL, respectively. Five of the strains examined increased the pH of the yogurt, while two strains were observed to reduce the pH. In milk, one strain raised the pH, while eleven strains reduced the pH. The levels of lactate, acetate, and formate in both the yogurt and milk were altered in a strain-specific manner. The results suggested that the metabolism by these strains differed significantly during the shelf-life. Careful strain selection is required to identify probiotic L. casei strains that will survive through shelf-life in either yogurt or fluid milk and not impact product quality.

Static and Dynamic Postural Characteristics in Patients with Chronic Obstructive Pulmonary Disease: The Relationship with Dyspnea and Pulmonary Functions.

BACKGROUND: There is little evidence about posture influence and its relationship with pulmonary functions in chronic obstructive pulmonary diseases(COPD) patients. OBJECTIVES: To compare spinal curvature, mobility, and postural competency in participants with and without COPD and investigate the relationship of postural characteristics with dyspnea and pulmonary functions in COPD patients. METHODS: We included 47 COPD patients and 47 age and gender-matched controls in our cross-sectional study. Participants underwent the following evaluations: modified Medical Research Council Dyspnea Scale, respiratory function test, and postural measurements in the sagittal plane in a standing position using a non-invasive, computer-assisted electromechanical device. Postural variables were compared between groups, and the relationship between postural variables with dyspnea and pulmonary functions was analyzed by multivariate regression analysis. RESULTS: Thoracic and lumbar curvature were higher (p<0.05), thoracic mobility and spinal inclination were lower (p=0.011, p=0.030, respectively) in patients with COPD. Thoracic angle and spinal inclination increased in COPD patients (p=0.040, p=0.011, respectively) while only spinal tilt increased in the control group (p=0.010) under spinal loading. Thoracic angle and mobility were related with dyspnea (r2=0.25, p<0.001), forced expiratory volume in the first second (r2=0.56, p<0.001), forced vital capacity (r2=0.41, p<0.001), and RV (r2 = 0.42, p<0.001). CONCLUSION: COPD patients had greater thoracic and lumbar angles in the static upright posture and lower thoracic mobility and spinal inclination in the sagittal plane. It was observed that patients increase their thoracic angles to maintain postural stability in dynamic conditions. Thoracic angle and mobility were related to dyspnea and pulmonary functions.

Turkish translation, cross-cultural adaptation, and assessment of psychometric properties of the Ottawa sitting scale for the intensive care unit survivors.

PURPOSE: To translate and cross-culturally adapt the Ottawa Sitting Scale (OSS) developed for acute care patients into Turkish and to examine its psychometric properties in intensive care unit (ICU) survivors. METHODS: After translation process the Turkish version of the scale was adminstered to eighty-one patients aged 39-82 years after discharge from the ICU and videotape was recorded. Two physiotherapists watched the videotaped records during the evaluation and scored. Test-retest reliability was assesed by scoring the same video recordings 15 days after the initial scoring. Correlations of the OSS with Berg Balance Scale (BBS), Functional Independence Measurement (FIM), the ICU length of stay and, total mechanical ventilation duration were assessed for convergent validity. RESULTS: The intra-class correlation coefficient for inter and intra-rater reliability was 0.989-0.994 and 0.998 respectively. The internal consistency was excellent (Cronbach's α = 0.998). The OSS score was highly correlated with the BBS total score (r = 0.716), the BBS unsupported sitting item score (r = 0.863), and moderately correlated with the total score of the FIM (r = 0.602), the number of days in ICU (r= -0.545), and total mechanical ventilation duration (r = -0.518). CONCLUSION: The Turkish version of OSS has been found to be valid and reliable in assessment of sitting balance in patients discharged from intensive care unit.Implications For RehabilitationPhysiotherapeutic measurements are crucial and necessary to determine the functional status of the patients.Ottawa Sitting Scale was translated and culturally adapted to Turkish and showed good psychometric propertiesOttawa Sitting Scale is valid and reliable tool to evaluate sitting balance in intensive care unit survivors.Ottawa Sitting Scale can guide clinicians in establishing evaluation and rehabilitation programs for patients discharged from intensive care unit.

Predictors of improvement in resting heart rate after exercise training in patients with chronic obstructive pulmonary disease.

BACKGROUND: High resting heart rate (RHR) is associated with multiple morbidity in chronic obstructive pulmonary disease (COPD) patients. Factors regarding the effectiveness of exercise training (ET) on RHR in COPD patients are unclear. AIMS: The main objective of the current study is to determine the predictors of the eventual change in RHR after ET. METHODS: One hundred and ten COPD patients (mean age: 63.1 ± 8.1 years, FEV1%: 43.6 ± 16.6) who participated in the ET program that consisted of supervised breathing, aerobic, strengthening, and stretching exercises for 8 weeks, 2 days a week, were included in the study. RHR, pulmonary functions, 6-min walk distance (6-MWD), Modified Medical Research Council Dyspnea Scale, St. George Respiratory Questionnaire, and Hospital Anxiety and Depression scores were compared before and after ET. Multivariate regression analysis was performed to correlate factors related to changes in RHR before and after exercise. RESULTS: There was a significant improvement in RHR after the ET program (p < 0.001). Improvement in RHR was correlated with baseline RHR, 6-MWD, partial arterial oxygen pressure, dyspnea sensation, forced expiratory volume in the first second (r = 0.516, -0.388, -0.489, 0.369, -0.360, p < 0.05, respectively), and change in 6-MWD, partial arterial oxygen pressure, and symptom score (r = 0.523, 0.451, -0.325, p < 0.05, respectively) after ET. Baseline RHR, 6-MWD, and the change in 6-MWD were the independent factors that predicted the change in RHR after ET. CONCLUSIONS: Patients with a high RHR and low functional capacity and whose functional capacity improves more have a greater decrease in RHR after the ET program. By considering these related factors, clinicians can focus on improving the cardiovascular system in COPD patients. CLINICAL TRIAL NUMBER: NCT04890080 (retrospectively registered-date of registration: 05.17.2021).

Neuropathy in COVID-19 associated with dysbiosis-related inflammation.

Although COVID-19 affects mainly lungs with a hyperactive and imbalanced immune response, gastrointestinal and neurological symptoms such as diarrhea and neuropathic pains have been described as well in patients with COVID-19. Studies indicate that gut-lung axis maintains host homeostasis and disease development with the association of immune system, and gut microbiota is involved in the COVID-19 severity in patients with extrapulmonary conditions. Gut microbiota dysbiosis impairs the gut permeability resulting in translocation of gut microbes and their metabolites into the circulatory system and induce systemic inflammation which, in turn, can affect distal organs such as the brain. Moreover, gut microbiota maintains the availability of tryptophan for kynurenine pathway, which is important for both central nervous and gastrointestinal system in regulating inflammation. SARS-CoV-2 infection disturbs the gut microbiota and leads to immune dysfunction with generalized inflammation. It has been known that cytokines and microbial products crossing the blood-brain barrier induce the neuroinflammation, which contributes to the pathophysiology of neurodegenerative diseases including neuropathies. Therefore, we believe that both gut-lung and gut-brain axes are involved in COVID-19 severity and extrapulmonary complications. Furthermore, gut microbial dysbiosis could be the reason of the neurologic complications seen in severe COVID-19 patients with the association of dysbiosis-related neuroinflammation. This review will provide valuable insights into the role of gut microbiota dysbiosis and dysbiosis-related inflammation on the neuropathy in COVID-19 patients and the disease severity.

Gut-lung axis and dysbiosis in COVID-19.

COVID-19, a novel infectious disease, caused by SARS-CoV-2, affected millions of people around the world with a high mortality rate. Although SARS-CoV-2 mainly causes lung infection, gastrointestinal symptoms described in COVID-19 patients and detection of the viral RNA in feces of infected patients drove attentions to a possible fecal-oral transmission route of SARS-CoV-2. However, not only the viral RNA but also the infectious viral particles are required for the viral infection and no proof has been demonstrated the transmission of the infectious virus particles via the fecal-oral route yet. Growing evidence indicates the crosstalk between gut microbiota and lung, that maintains host homeostasis and disease development with the association of immune system. This gut-lung interaction may influence the COVID-19 severity in patients with extrapulmonary conditions. Severity of COVID-19 has mostly associated with old ages and underlying medical conditions. Since the diversity in the gut microbiota decreases during aging, dysbiosis could be the reason for older adults being at high risk for severe illness from COVID-19. We believe that gut microbiota contributes to the course of COVID-19 due to its bidirectional relationship with immune system and lung. Dysbiosis in gut microbiota results in gut permeability leading to secondary infection and multiple organ failure. Conversely, disruption of the gut barrier integrity due to dysbiosis may lead to translocation of SARS-CoV-2 from the lung into the intestinal lumen via circulatory and lymphatic system. This review points out the role of dysbiosis of the gut microbiota involving in sepsis, on the severity of SARS-CoV-2 infection. Additionally, this review aims to clarify the ambiguity in fecal-oral transmission of SARS- CoV-2.

Biochemical and Molecular Characterizations of a Novel pH- and Temperature-Stable Pectate Lyase from Bacillus amyloliquefaciens S6 for Industrial Application.

In this paper, we report cloning of a pectate lyase gene from Bacillus amyloliquefaciens S6 (pelS6), and biochemical characterization of the recombinant pectate lyase. PelS6 was found to be identical with B. subtilis 168 pel enzyme with 100% amino acid sequence homology. Although these two are genetically very close, they are distinctly different in physiology. pelS6 gene encodes a 421-aa protein with a molecular mass of 65,75 kDa. Enzyme activity increased from 12.8 ± 0.3 to 49.6 ± 0.4 units/mg after cloning. The relative enzyme activity of the recPel S6 ranged from 80% to 100% at pH between 4 and 14. It was quite stable at different temperature values ranging from 15 to 90 °C. The recPEL S6 showed a maximal activity at pH 10 and at 60 °C. 0.5 mM of CaCl2 is the most effective metal ion on the recPEL S6 as demonstrated by its increased relative activity with 473%. recPEL S6 remained stable at - 20 °C for 18 months. In addition recPEL S6 increased juice clarity. This study introduces a novel bacterial pectate lyase enzyme with its characteristic capability of being highly thermostable, thermotolerant, and active over a wide range of pH, meaning that it can work at both acidic and alkaline environments, which are the most preferred properties in the industry.

The relationship between the structural characteristics of lactobacilli-EPS and its ability to induce apoptosis in colon cancer cells in vitro.

Colon cancer is one of the most common cancer around the world. Exopolysaccharides (EPSs) produced by lactobacilli as potential prebiotics have been found to have an anti-tumor effect. In this study, lyophilized EPSs of four Lactobacillus spp. for their impact on apoptosis in colon cancer cells (HT-29) was evaluated using flow cytometry. The relationship between capability of a lactobacilli-EPS to induce apoptosis and their monosaccharide composition, molecular weight (MW), and linkage type was investigated by HPLC, SEC, and NMR, respectively. Changes in apoptotic-markers were examined by qPCR and Western Blotting. EPSs were capable of inhibiting proliferation in a time-dependent manner and induced apoptosis via increasing the expression of Bax, Caspase 3 and 9 while decreasing Bcl-2 and Survivin. All EPSs contained mannose, glucose, and N-acetylglucosamine with different relative proportions. Some contained arabinose or fructose. MW ranged from 102-104Da with two or three fractions. EPS of L. delbrueckii ssp. bulgaricus B3 having the highest amount of mannose and the lowest amount of glucose, showed the highest apoptosis induction. In conclusion, lactobacilli-EPSs inhibit cell proliferation in HT-29 via apoptosis. Results suggest that a relationship exists between the ability of EPS to induce apoptosis and its mannose and glucose composition.

Impact of Exopolysaccharides (EPSs) of Lactobacillus gasseri strains isolated from human vagina on cervical tumor cells (HeLa).

Lactobacilli, commonly used as probiotics, have been shown to maintain vaginal health and contribute to host microbiota interaction. Exopolysaccharides (EPSs) produced by lactobacillus have been found to have an important role in probiotic activity; however, there is limited knowledge concerning their impact on cervical cancer and urogenital health. The objective of this study is to investigate and compare EPSs of L. gasseri strains (G10 and H15), isolated from a healthy human vagina, for their capability to inhibit cervical cancer cell (HeLa) growth and modulate immune response. HeLa cells were treated with live culture at ∼108 CFU/ml or increasing concentration of lyophilized EPS (L-EPS) (100, 200, or 400 µg/ml) of L. gasseri strains and their ability to adhere to host cells, inhibit proliferation, and modulate immune response were evaluated. Additionally, monosaccharide composition of the L-EPSs produced by L. gasseri strains was determined by HPLC. The sugar component was the same; however, relative proportions of the individual monosaccharides except mannose were different. Although they both produce similar amount of EPS, the most adhesive strain was G10. Both live and L-EPS of L. gasseri strains were capable of inhibiting the cell proliferation of HeLa cells with the impact of L-EPS being strain specific. L-EPSs of L. gasseri strains induced apoptosis in HeLa cells in a strain dependent manner. The ability to induce apoptosis by G10 associated with an upregulation of Bax and Caspase 3. L. gasseri strains showed an anti-inflammatory impact on HeLa cells by decreasing the production of TNF-α and increasing the IL-10 production. In conclusion, diversity in sugar composition of EPS might contribute to adhesion and proliferation properties. Although our results suggest a relationship between the ability of a strain to induce apoptosis and its sugar composition of EPS, further research is required to determine the probiotic mechanisms of action by which L. gasseri strains result in strain specific anti-proliferative activity.

The Impact of Lactobacillus casei on the Composition of the Cecal Microbiota and Innate Immune System Is Strain Specific.

The probiotic function to impact human health is thought to be related to their ability to alter the composition of the gut microbiota and modulate the human innate immune system. The ability to function as a probiotic is believed to be strain specific. Strains of Lactobacillus casei are commonly utilized as probiotics that when consumed alter the composition of the gut microbiota and modulate the host immune response. L. casei strains are known to differ significantly in gene content. The objective of this study was to investigate seven different L. casei strains for their ability to alter the murine gut microbiota and modulate the murine immune system. C57BL/6 mice were fed L. casei strains at a dose of 108 CFU/day/mouse for seven days and sacrificed 3.5h after the last administration. The cecal content and the ileum tissue were collected for microbiota analysis and immune profiling, respectively. While 5 of the L. casei strains altered the gut microbiota in a strain specific manner, two of the strains did not alter the overall cecal microbiota composition. The observed changes cluster into three groups containing between 1 and 2 strains. Two strains that did not affect the gut microbiota composition cluster together with the control in their impact on pattern recognition receptors (PRRs) expression, suggesting that the ability to alter the cecal microbiota correlates with the ability to alter PRR expression. They also cluster together in their impact on the expression of intestinal antimicrobial peptides (AMPs). This result suggests that a relationship exists between the capability of a L. casei strains to alter the composition of the gut microbiota, PRR regulation, and AMP regulation.

The Effect of Lactobacillus casei 32G on the Mouse Cecum Microbiota and Innate Immune Response Is Dose and Time Dependent.

Lactobacilli have been associated with a variety of immunomodulatory effects and some of these effects have been related to changes in gastrointestinal microbiota. However, the relationship between probiotic dose, time since probiotic consumption, changes in the microbiota, and immune system requires further investigation. The objective of this study was to determine if the effect of Lactobacillus casei 32G on the murine gastrointestinal microbiota and immune function are dose and time dependent. Mice were fed L. casei 32G at doses of 106, 107, or 108 CFU/day/mouse for seven days and were sacrificed 0.5h, 3.5h, 12h, or 24h after the last administration. The ileum tissue and the cecal content were collected for immune profiling by qPCR and microbiota analysis, respectively. The time required for L. casei 32G to reach the cecum was monitored by qPCR and the 32G bolus reaches the cecum 3.5h after the last administration. L. casei 32G altered the cecal microbiota with the predominance of Lachnospiraceae IS, and Oscillospira decreasing significantly (p < 0.05) in the mice receiving 108 CFU/mouse 32G relative to the control mice, while a significant (p < 0.05) increase was observed in the prevalence of lactobacilli. The lactobacilli that increased were determined to be a commensal lactobacilli. Interestingly, no significant difference in the overall microbiota composition, regardless of 32G doses, was observed at the 12h time point. A likely explanation for this observation is the level of feed derived-nutrients resulting from the 12h light/dark cycle. 32G results in consistent increases in Clec2h expression and reductions in TLR-2, alpha-defensins, and lysozyme. Changes in expression of these components of the innate immune system are one possible explanation for the observed changes in the cecal microbiota. Additionally, 32G administration was observed to alter the expression of cytokines (IL-10rb and TNF-α) in a manner consistent with an anti-inflammatory response.

Glycomacropeptide is a prebiotic that reduces Desulfovibrio bacteria, increases cecal short-chain fatty acids, and is anti-inflammatory in mice.

Glycomacropeptide (GMP) is a 64-amino acid (AA) glycophosphopeptide with application to the nutritional management of phenylketonuria (PKU), obesity, and inflammatory bowel disease (IBD). GMP is a putative prebiotic based on extensive glycosylation with sialic acid, galactose, and galactosamine. Our objective was to determine the prebiotic properties of GMP by characterizing cecal and fecal microbiota populations, short-chain fatty acids (SCFA), and immune responses. Weanling PKU (Pah(enu2)) and wild-type (WT) C57Bl/6 mice were fed isoenergetic AA, GMP, or casein diets for 8 wk. The cecal content and feces were collected for microbial DNA extraction to perform 16S microbiota analysis by Ion Torrent PGM sequencing. SCFA were determined by gas chromatography, plasma cytokines via a Bio-Plex Pro assay, and splenocyte T cell populations by flow cytometry. Changes in cecal and fecal microbiota are primarily diet dependent. The GMP diet resulted in a reduction from 30-35 to 7% in Proteobacteria, genera Desulfovibrio, in both WT and PKU mice with genotype-dependent changes in Bacteroidetes or Firmicutes. Cecal concentrations of the SCFA acetate, propionate, and butyrate were increased with GMP. The percentage of stimulated spleen cells producing interferon-γ (IFN-γ) was significantly reduced in mice fed GMP compared with casein. In summary, plasma concentrations of IFN-γ, TNF-α, IL-1ß, and IL-2 were reduced in mice fed GMP. GMP is a prebiotic based on reduction in Desulfovibrio, increased SCFA, and lower indexes of inflammation compared with casein and AA diets in mice. Functional foods made with GMP may be beneficial in the management of PKU, obesity, and IBD.