Ethanolic extract of Euphorbia royleana Boiss. reduces metastasis of breast cancer cells and inhibits tumor progression in vivo.

Metastasis is the most devastating attribute of breast cancer (BC) that leads to high mortality. It is a complex process of tumor cell migration, invasion, and angiogenesis. In this study, we evaluated the effect of ERA on BC metastasis and BC progression in vivo. The transwell invasion/migration and wound healing assays showed that ERA treatment significantly reduced the invasion and migration of BC cell lines. The expression of mesenchymal (E-cadherin and N-cadherin), matrix metalloproteinases (MMP2, MMP9), and stemness markers (Oct3) were down-regulated by ERA. Furthermore, ERA down-regulated angiogenic chemokines (CXCL1/2/3, CXCL5, and CXCL12) expression in the highly metastatic MDA-MB-231 cell line. The clonogenic survival of BC cells was also reduced by ERA treatment. Strikingly, ERA prevented DMBA-induced tumor growth in Swiss albino mice as depicted by a high animal survival rate (84%) in the ERA group and histopathological analysis. Conclusively, this study revealed that ERA possesses anti-metastatic potential and also reduces the growth of BC in vivo. Moreover, the GC-MS data revealed the presence of biologically active compounds (Lupeol, Phytol, phytosterol) and some rare (9, 19-Cyclolanost) phyto metabolites in ERA extract. However, further studies are suggestive to identify and isolate the therapeutic agents from ERA to combat BC and metastasis.

Serological and epidemiological investigation of Mycobacterium avium subspecies paratuberculosis in bovines in Pakistan.

Objective: To investigate the prevalence of paratuberculosis in cattle and buffaloes at twelve public dairy farms in Punjab, Pakistan. Methods: A total of 2181 more than two-year-old animals (1242 cattle and 939 buffaloes) were tested by avian tuberculin, i.e., killed purified protein derivative (PPD) of Mycobacterium avium paratuberculosis and indirect ELISA. Blood and fecal samples were collected from tuberculin positive animals. These samples were further processed by indirect ELISA. The data were analyzed using frequency analysis and logistic analysis procedures. Results: The prevalence of paratuberculosis at public dairy farms was 3.8%, as determined by tuberculin + ELISA test. It varied from 0.71-13.5% with a 100% herd prevalence. Multivariate logistic regression analysis revealed that species, milk production, total animals, total small ruminants, and total buffaloes were significantly associated with the occurrence of paratuberculosis. Odd ratio analysis revealed that with a one-kilogram increase in body weight, there will be a 0.006% increase in disease occurrence. With the increase in one animal in small ruminants and buffaloes, there will be 0.008% and 0.42% greater chances of developing paratuberculosis, respectively. Bivariate logistic regression analysis of cattle and buffaloes revealed that farm number, age, and total number of cattle were significantly associated with the occurrence of paratuberculosis. A one-month increase in lactation length increases the chance of tuberculosis by 0.004%; similarly, a one-liter increase in milk production increases the chance of disease by 10%. With each additional buffalo in the herd, there will be a 0.007% greater chance for the occurrence of paratuberculosis. Conclusion: This study concluded that tuberculin testing can be used in conjunction with ELISA to screen animals for paratuberculosis in countries with scarce resources, such as Pakistan. The efficacy of disease diagnosis can be improved by combining multiple tests.

Factors Associated With Short-Term Complications After Percutaneous Endoscopic Gastrostomy Tube Insertion: A Retrospective Cohort Study.

BACKGROUND: Percutaneous endoscopic gastrostomy (PEG) tube placement is generally safe but is associated with a range of complications. Minor complications include infections, granuloma formation, leakage, and blockages, while major complications encompass aspiration pneumonia, hemorrhage, and more serious conditions such as necrotizing fasciitis and colonic fistula. AIM: This study aimed to assess the rate of short-term complications within one month of endoscopic PEG insertion, focusing on their correlation with patient characteristics. METHODOLOGY: This retrospective cohort study analyzed data from patients who underwent PEG insertion between January 2020 and December 2022. It evaluated the incidence of complications in relation to variables such as the indication for the procedure, the patient's immune status, albumin and CRP levels, and the setting of the procedure (inpatient vs. outpatient). RESULTS: The study included 121 patients, with a mean age of 69.73 years, comprising 71 males (58.7%) and 50 females (41.3%). Neurological indications accounted for 64.5% of the cases. Notably, 67.8% of the patients were immunocompromised. Within 30 days of PEG insertion, 16.5% experienced complications, including GI bleeding (4.1%), infection at the PEG site (11.6%), and peritonitis (0.8%). Complications were significantly higher in immunocompromised patients and those with non-neurological indications. Higher serum albumin and lower CRP levels were associated with fewer complications, though the association was not statistically significant. CONCLUSION: The study highlights that gastrostomy site infection is the most common short-term complication following PEG insertion. Immune status and the reason for PEG insertion emerged as key factors influencing the likelihood of complications.

Personalized Multimodal Lifestyle Intervention as the Best-Evidenced Treatment for Chronic Pain: State-of-the-Art Clinical Perspective.

Chronic pain is the most prevalent disease worldwide, leading to substantial disability and socioeconomic burden. Therefore, it can be regarded as a public health disease and major challenge to scientists, clinicians and affected individuals. Behavioral lifestyle factors, such as, physical (in)activity, stress, poor sleep and an unhealthy diet are increasingly recognized as perpetuating factors for chronic pain. Yet, current management options for patients with chronic pain often do not address lifestyle factors in a personalized multimodal fashion. This state-of-the-art clinical perspective aims to address this gap by discussing how clinicians can simultaneously incorporate various lifestyle factors into a personalized multimodal lifestyle intervention for individuals with chronic pain. To do so the available evidence on (multimodal) lifestyle interventions targeting physical (in)activity, stress, sleep and nutritional factors, specifically, was reviewed and synthetized from a clinical point of view. First, advise is provided on how to design a personalized multimodal lifestyle approach for a specific patient. Subsequently, best-evidence recommendations on how to integrate physical (in)activity, stress, sleep and nutritional factors as treatment targets into a personalized multimodal lifestyle approach are outlined. Evidence supporting such a personalized multimodal lifestyle approach is growing, but further studies are needed.

Effects of robot-assisted arm training on respiratory muscle strength, activities of daily living, and quality of life in patients with stroke: a single-blinded randomized controlled trial.

BACKGROUND: Post-stroke clinical changes not only affect extremities and trunk muscles but also the respiratory muscles. PURPOSE: To determine the effect of robot-assisted arm training with conventional rehabilitation (CombT) on respiratory muscle strength, activities of daily living (ADL), and quality of life in patients with stroke and to compare the results with conventional rehabilitation (CR). METHODS: It was a two-arm, single-blinded, randomized controlled trial in which 66 patients were randomly allocated to either CombT or CR to receive 30 sessions (5/week) over 6 weeks. The respiratory muscle strength (maximal inspiratory pressure (MIP) and maximal expiratory pressure (MEP)), activities of daily life (ABILHAND questionnaire), and quality of life (Stroke Impact Scale (SIS)) were measured before and 6 weeks after training. RESULTS: The CombT group showed significantly better MIP, MEP, and performance in ADLs after 6 weeks of training compared to the CR group (p < .01). The effect size was large for MIP (d = 0.9) and MEP (d = 0.9), whereas medium for performance in ADLs (d = 0.62). Also, the SIS-arm strength (p < .01), hand function (p = .04), ADLs (p = .02), and recovery (p = .04) were significantly better in CombT group with a medium (d = 0.6, d = 0.5, d = 0.5, and d = 0.5, respectively) effect size compared with CR group. CONCLUSIONS: Both CombT and CR groups improved respiratory muscle strength, performance in ADLs, and quality of life in patients with stroke. However, CombT appears to offer more comprehensive benefits, highlighting its valuable role in respiratory and functional recovery after stroke.

The effects of low-intensity resistance training with blood flow restriction versus traditional resistance exercise on lower extremity muscle strength and motor functionin ischemic stroke survivors: a randomized controlled trial.

BACKGROUND: Blood flow restriction (BFR) training can temporarily reduce cortical GABA concentrations and increase the size of motor volleys to deafferented muscles, which can promote motor recovery in stroke survivors. OBJECTIVE: To determine the effect of low-intensity resistance training with BFR (LIRT-BFR) on lower extremity muscle strength, balance, functional mobility, walking capacity, gait speed, anxiety, and depression in stroke survivors and to compare the results with high-intensity resistance training (HIRT). METHOD: It was a two-arm, single-blinded, randomized controlled trial in which 32 ischemic stroke participants were randomly allocated to LIRT-BFR or HIRT group. The LIRT-BFR group received low load resistance training (40% of 1-Repetition Maximum (1-RM)) with BFR, whereas HIRT group received high load resistance training (80% of 1-RM). The 6-Minute Walk Test (6-MWT), five-time sit-to-stand test (5TSTST), Timed Up and Go (TUG) test, and Barthel index were the primary outcome measures. The secondary outcome measures included gait speed (m/s), stride length (cm), cadence (steps/min), and Hospital Anxiety and Depression. RESULTS: All the primary and secondary outcome measures were significantly improved in both groups (p < 0.05). The LIRT-BFR group showed a slightly greater, but non-significant, improvement as compared to the HIRT group in terms of mean change observed in 6-MWT (81 m vs 62 m), 5TSTST (-5.27 vs -4.81), gait speed (0.19 vs 0.12), stride length (18 vs 13), and cadence (8 vs 6). No adverse event was reported. CONCLUSION: LIRT-BFR produced a significant improvement in muscle strength, balance, walking capacity, and anxiety and depression in ischemic stroke patients, and the improvement are comparable to HIRT. CLINICAL TRIAL REGISTRATION: NCT05281679.

Seroprevalence and haemato-biochemical effects of bovine leucosis in buffalo, Punjab, Pakistan.

Enzootic bovine leucosis is caused by bovine leukaemia virus (BLV), a Deltaretrovirus belonging to the family Retroviridae. BLV causes huge economic losses to the dairy industry in the form of decreased milk production, premature culling, and poor reproductive performance of the animals. The aim of the present study was to determine the seroprevalence of BLV infection in buffalo in two districts of Punjab, Pakistan. A total of 384 samples were collected and analysed using a commercial indirect enzyme-linked immunosorbent assay (ELISA) to investigate the seroprevalence of BLV through the detection of the anti-BLV gp51 antibody. A predesigned data questionnaire proforma was employed to find out the association of risk factors with disease. Overall, 18.2% of buffaloes were seropositive for BLV in the study population. The results revealed a significant association (P < 0.05) of age with BLV infection. Furthermore, milk yield and pregnancy had a significant association with the seroprevalence of BLV infection in buffalo whereas no significant association was found with sex, breeding, and health status. Biochemical and oxidative stress markers revealed a significant decrease in liver enzymes alanine transaminase (ALT) and aspartate transaminase (AST), glutathione peroxidase (GPX), and superoxide dismutase (SOD) in seropositive animals as compared to healthy animals. It is concluded that BLV has a considerable prevalence in buffalo in Punjab, Pakistan and there is a dire need to investigate the disease epidemiology at both national and international levels and strategies should be developed to implement an effective control program.

Corrigendum: Sero-epidemiological study of zoonotic bacterial abortifacient agents in small ruminants.

[This corrects the article DOI: 10.3389/fvets.2023.1195274.].

A catch-and-release nano-based gene delivery system.

The design of nanomaterial-based nucleic acid formulations is one of the biggest endeavours in the search for clinically applicable gene delivery systems. Biopolymers represent a promising subclass of gene carriers due to their physicochemical properties, biodegradability and biocompatibility. By modifying melanin-like polydopamine nanoparticles with poly-L-arginine and poly-L-histidine blends, we obtained a novel catch-and-release gene delivery system for efficient trafficking of pDNA to human cells. A synergistic interplay of nanoparticle-bound poly-L-arginine and poly-L-histidine was observed and evaluated for pDNA binding affinity, cell viability, gene release and transfection. Although the functionalisation with poly-L-arginine was crucial for pDNA binding, the resulting nanocarriers failed to release pDNA intracellularly, resulting in limited protein expression. However, optimal pDNA release was achieved through the co-formulation with poly-L-histidine, essential for pDNA release. This effect enabled the design of gene delivery systems, which were comparable to Lipofectamine in terms of transfection efficacy and the catch-and-release surface modification strategy can be translated to other nanocarriers and surfaces.

Corrigendum: From soil to plant: strengthening carrot defenses against Meloidogyne incognita with vermicompost and arbuscular mycorrhizal fungi biofertilizers.

[This corrects the article DOI: 10.3389/fmicb.2023.1206217.].

Molecular identification and infection pathology of Mycobacterium spp. in captive wild animals in Pakistan.

INTRODUCTION: Animal tuberculosis is an infectious, chronic, granulomatous, and debilitating disease affecting animals as well as humans. However, in recent decades, there have been many endemic geographic localities where animal tuberculosis has been identified in wildlife reservoirs, limiting the eradication program in cattle. This study aimed to identify animal tuberculosis in captive zoo animals in Pakistan. METHODOLOGY: In total, 185 morbid zoo animals were brought for postmortem examination at a veterinary postmortem facility. During the macroscopic examination, these animals were thoroughly examined for the presence of suggestive gross lesions of animal tuberculosis (granulomas/tubercles), and the pattern and distribution of these lesions in different organs. The Ziehl-Neelsen (ZN) staining was performed on smears prepared from granulomatous lesions of lung tissue followed by molecular identification of M. bovis and M. tuberculosis DNA using polymerase chain reaction (PCR). RESULTS: The postmortem examination revealed that 8.1% (15/185) of animals had gross tuberculosis lesions on the lungs and lymph nodes. The ZN staining of tissue smears showed 5.40% positivity while M. bovis and M. tuberculosis DNA was identified in 3.78 % and 1.1% of investigated animals, respectively. CONCLUSIONS: The study showed that animal tuberculosis is prevalent among wildlife in Pakistan and it may pose serious public health concerns to the people visiting these zoos and wildlife parks.

Sero-epidemiological study of zoonotic bacterial abortifacient agents in small ruminants.

Abortion is one of the leading causes of economic losses in the livestock industry worldwide. Chlamydia abortus, Coxiella burnetii, and Brucella spp. are the leading cause of late fetal loss in small ruminants. This study determined the seroprevalence of these agents in small ruminants in district Jhang. A total of 385 serum samples were taken from the sheep and goats from different flocks with a history of abortion and subjected to i-ELISA. Further, samples were analysed for liver enzymes and total serum protein using a semi-automated chemistry analyzer. The result of indirect commercial ELISA showed 13.0, 4.2 and 11.2% prevalence for Coxiella burnetii, Chlamydia abortus, and Brucella spp., respectively. Values of different serum parameters (ALT, AST, and total protein) of seropositive animals were also determined. There was a significant rise in AST and ALT values of infected animals (p ≤ 0.05). Total protein decreased for all three infections, but a significant drop was noted in Brucella positive sheep serum samples. Various risk factors were studied. Binary logistic regression proved a significant role of ticks for coxiellosis and brucellosis. Age, parity, and species did not impact the prevalence of diseases studied.

From soil to plant: strengthening carrot defenses against Meloidogyne incognita with vermicompost and arbuscular mycorrhizal fungi biofertilizers.

Introduction: Sustainable agricultural practices for controlling crop pests are urgently needed to reduce the reliance on chemical pesticides, which have long-term detrimental effects on ecosystems. In this study, we assessed the effectiveness of arbuscular mycorrhizal fungi (AMF) and vermicompost (Vc) supplementation, alone and in combination, in mitigating the negative impacts of Meloidogyne incognita infestation on carrot (Daucus carota L.) growth, development, and physiology. Methods: We measured different plant growth parameters such as plant height and biomass accumulation, several plant physiological parameters such as the levels of photosynthetic pigments, phenolics, and the activity of defense enzymes such as peroxidases and polyphenol oxidases, and evaluated the severity of Meloidogyne incognita nematode infestation on plants treated or not treated with vermicompost (Vc) and/or arbuscular mycorrhizal fungi (AMF). Results: Our findings show that M. incognita significantly affects plant growth, biomass accumulation, and photosynthetic pigment and carotenoid content. The incorporation of Vc and AMF into the soil, either individually or in combination, significantly alleviates the negative effects of nematode infestation on carrot plants. This was accompanied by the induction of phenolic compounds and defense enzymes such as peroxidases (+15.65%) and polyphenol oxidases (29.78%), and by a reduction in the severity of nematode infestation on Vc and AMF-treated plants compared to nematode-infested plants. Principal component analysis (PCA) shows significant correlations between various of the studied parameters. In particular, we observed negative correlations between the application of AMF and Vc alone and in combination and disease severity, and positive correlations between plant growth, photosynthetic pigments phenol content, and activity of defense enzymes. Discussion: Our study highlights the relevance of cultural practices and beneficial microorganisms for the sustainable and environmentally friendly management of agricultural pests.

Imine Derivatives of Benzoxazole Attenuate High-Fat Diet-Induced Hyperlipidemia by Modulation of Lipid-Regulating Genes.

Purpose: Hyperlipidemia being the prominent risk factor of cardiovascular diseases and side effects associated with the current lipid-lowering drugs have attracted the interest of scientists in the quest for new alternatives. In view of the diverse pharmacological potentials of benzoxazole (BZX) compounds, this study was designed to evaluate the antihyperlipidemic activity of imine derivatives of BZX in high-fat diet (HFD)-fed rats. Methods: Hyperlipidemia was induced in Sprague-Dawley rats by using HFD for 28 days. On the 28th day, blood samples were collected, and animals having serum triglycerides (TG) greater than 400 mg/dL and total cholesterol (TC) greater than 280 mg/dL were selected for further study. Hyperlipidemic rats were daily treated with either a vehicle or simvastatin (SIM; 20 mg/kg) or BZX compounds (10, 20, and 30 mg/kg), for 12 consecutive days. After the specified time duration, hyperlipidemic biomarkers were evaluated in the blood samples of sacrificed rats. Liver samples were collected for histopathological and mRNA analyses. Binding affinities of BZX derivatives with different targets were assessed by molecular docking. Results: The present study revealed that the BZX derivatives dose-dependently reduced the serum levels of TC, TG, low-density lipoprotein, and very low-density lipoprotein along with improvement in high-density lipoprotein levels. Similarly, all the compounds reduced HFD-induced alanine transaminase and aspartate aminotransferase levels except BZX-4. Histopathology of liver samples demonstrated mild to moderate fatty changes upon treatment with BZX-1, BZX-2, and BZX-4. The hepatic architecture of the BZX-3-treated samples was close to normal, and only mild inflammation was witnessed in these samples. Moreover, all the compounds significantly increased superoxide dismutase and glutathione levels, indicating their antioxidant potentials. Gene expression data showed that BZX-1 and BZX-3 reduced lipid levels by inhibiting HMGCR, APOB, PCSK9, SRB1, and VCAM1 and via improving PPAR-α and APOE mRNA levels. BZX-2 demonstrated its antihyperlipidemic effects mainly due to inhibition of APOB, while BZX-4-mediated effects appeared to be due to attenuation of APOB, PCSK9, and SRB1. BZX derivatives displayed strong binding affinities with HMGCR, APOB, and VCAM1, which suggested that some of the interactions might be required for inhibition of these target proteins. Conclusions: Based on the current findings, it can be concluded that BZX derivatives exert their antihyperlipidemic effects via modulation of multiple lipid-regulating genes.

Non-invasive Brain Stimulation Techniques for the Improvement of Upper Limb Motor Function and Performance in Activities of Daily Living After Stroke: A Systematic Review and Network Meta-analysis.

OBJECTIVE: To compare the efficacy of non-invasive brain stimulation (NiBS) such as transcranial direct current stimulation (tDCS), repetitive transcranial magnetic stimulation (rTMS), theta-burst stimulation (TBS), and transcutaneous vagus nerve stimulation (taVNS) in upper limb stroke rehabilitation. DATA SOURCES: PubMed, Web of Science, and Cochrane databases were searched from January 2010 to June 2022. DATA SELECTION: Randomized controlled trials (RCTs) assessing the effects of "tDCS", "rTMS", "TBS", or "taVNS" on upper limb motor function and performance in activities of daily livings (ADLs) after stroke. DATA EXTRACTION: Data were extracted by 2 independent reviewers. Risk of bias was evaluated with the Cochrane Risk of Bias tool. DATA SYNTHESIS: 87 RCTs with 3750 participants were included. Pairwise meta-analysis showed that all NiBS except continuous TBS (cTBS) and cathodal tDCS were significantly more efficacious than sham stimulation for motor function (standardized mean difference [SMD] range 0.42-1.20), whereas taVNS, anodal tDCS, and both low and high frequency rTMS were significantly more efficacious than sham stimulation for ADLs (SMD range 0.54-0.99). NMA showed that taVNS was more effective than cTBS (SMD:1.00; 95% CI (0.02-2.02)), cathodal tDCS (SMD:1.07; 95% CI (0.21-1.92)), and Physical rehabilitation alone (SMD:1.46; 95% CI (0.59-2.33)) for improving motor function. P-score found that taVNS is best ranked treatment in improving motor function (SMD: 1.20; 95% CI (0.46-1.95)) and ADLs (SMD:1.20; 95% CI (0.45-1.94)) after stroke. After taVNS, excitatory stimulation protocols (intermittent TBS, anodal tDCS, and HF-rTMS) are most effective in improving motor function and ADLs after acute/sub-acute (SMD range 0.53-1.63) and chronic stroke (SMD range 0.39-1.16). CONCLUSIONS: Evidence suggests that excitatory stimulation protocols are the most promising intervention in improving upper limb motor function and performance in ADLs. taVNS appeared to be a promising intervention for stroke patients, but further large RCTs are required to confirm its relative superiority.

Quantitative assessment of present and the future potential threat of coastal erosion along the Odisha coast using geospatial tools and statistical techniques.

The eastern coast of India is one of the regions where most of the population resides in urban areas in the low-elevation coastal zone, making it vulnerable to frequent extreme weather events. The objectives of this study are to assess the short- to long-term shoreline changes of the Odisha coast, to understand how anthropogenic influences, and particularly extreme natural events, affect these changes, and to predict shoreline changes for 2050. This study utilized multi-temporal/spectral/spatial resolution satellite images and a digital shoreline analysis (DSAS) tool to appraise the short- (at five/six-year intervals) and long-term (1990-2019) shoreline dynamics along the coastal part of Odisha over the past three decades (1990-2019). The long-term shoreline analysis shows that the mean shoreline change is about 0.67 m/year and highlights that 52.47 % (227.4 km), 34.70 % (150.4 km), and 12.83 % (55.6 km) of the total Odisha coastline exhibit erosion, accretion, and stability, respectively. During the short-term analysis, the 2000-2005 period had the highest percentage of erosion (64.27 %), followed by the 2005-2010 period with an erosional trend of 59.06 %. The 1995-2000 period showed an accretion trend, whereas, during the last period, i.e., 2015-2019, the percentage of transects depicting erosion and accretion was almost similar. In 2050, 55.85 % of the transects are expected to show accretion, while 44.15 % would show erosion or a constant trend. The study identified the hotspots of coastal erosion along delineated study zones by synthesizing data from previous studies as well. The regional analysis of shoreline change along the Odisha coast would not only provide coastal managers with critical information on shoreline dynamics but also draw attention to vulnerable areas linked to shoreline dynamicity along the coast.