An Empirical Analysis of Sacha Inchi (Plantae: Plukenetia volubilis L.) Seed Proteins and Their Applications in the Food and Biopharmaceutical Industries.

Sacha Inchi (Plukenetia volubilis L.) is a plant native in the Amazon rainforest in South America known for its edible seeds, which are rich in lipids, proteins, vitamin E, polyphenols, minerals, and amino acids. Rural communities in developing nations have been using this plant for its health benefits, including as a topical cream for rejuvenating and revitalising skin and as a treatment for muscle pain and rheumatism. Although Sacha Inchi oil has been applied topically to soften skin, treat skin diseases, and heal wounds, its protein-rich seeds have not yet received proper attention for extensive investigation. Proteins in Sacha Inchi seeds are generally known to have antioxidant and antifungal activities and are extensively used nowadays in making protein-rich food alternatives worldwide. Notably, large-scale use of seed proteins has begun in nanoparticle and biofusion technologies related to the human health-benefitting sector. To extract and identify their proteins, the current study examined Sacha Inchi seeds collected from the Malaysian state of Kedah. Our analysis revealed a protein concentration of 73.8 ± 0.002 mg/g of freeze-dried seed flour. Employing liquid chromatography-tandem mass spectrometry (LC-MS/MS) and PEAKS studio analysis, we identified 217 proteins in the seed extract, including 152 with known proteins and 65 unknown proteins. This study marks a significant step towards comprehensively investigating the protein composition of Sacha Inchi seeds and elucidating their potential applications in the food and biopharmaceutical sectors. Our discoveries not only enhance our knowledge of Sacha Inchi's nutritional characteristics but also pave the way for prospective research and innovative advancements in the realms of functional food and health-related domains.

Systemic interleukin-6 inhibition ameliorates acute neuropsychiatric phenotypes in a murine model of acute lung injury.

Acute neuropsychiatric impairments occur in over 70% of patients with acute lung injury. Mechanical ventilation is a well-known precipitant of acute lung injury and is strongly associated with the development of acute delirium and anxiety phenotypes. In prior studies, we demonstrated that IL-6 mediates neuropathological changes in the frontal cortex and hippocampus of animals with mechanical ventilation-induced brain injury; however, the effect of systemic IL-6 inhibition on structural and functional acute neuropsychiatric phenotypes is not known. We hypothesized that a murine model of mechanical ventilation-induced acute lung injury (VILI) would induce neural injury to the amygdala and hippocampus, brain regions that are implicated in diverse neuropsychiatric conditions, and corresponding delirium- and anxiety-like functional impairments. Furthermore, we hypothesized that these structural and functional changes would reverse with systemic IL-6 inhibition. VILI was induced using high tidal volume (35 cc/kg) mechanical ventilation. Cleaved caspase-3 (CC3) expression was quantified as a neural injury marker and found to be significantly increased in the VILI group compared to spontaneously breathing or anesthetized and mechanically ventilated mice with 10 cc/kg tidal volume. VILI mice treated with systemic IL-6 inhibition had significantly reduced amygdalar and hippocampal CC3 expression compared to saline-treated animals and demonstrated amelioration in acute neuropsychiatric behaviors in open field, elevated plus maze, and Y-maze tests. Overall, these data provide evidence of a pathogenic role of systemic IL-6 in mediating structural and functional acute neuropsychiatric symptoms in VILI and provide preclinical justification to assess IL-6 inhibition as a potential intervention to ameliorate acute neuropsychiatric phenotypes following VILI.

Signature Effects of Vector-Guided Systemic Nano Bioconjugate Delivery Across Blood-Brain Barrier of Normal, Alzheimer's, and Tumor Mouse Models.

The ability to cross the blood-brain barrier (BBB) is critical for targeted therapy of the central nerve system (CNS). Six peptide vectors were covalently attached to a 50 kDa poly(ß-l-malic acid)-trileucine polymer forming P/LLL(40%)/vector conjugates. The vectors were Angiopep-2 (AP2), B6, Miniap-4 (M4), and d-configurated peptides D1, D3, and ACI-89, with specificity for transcytosis receptors low-density lipoprotein receptor-related protein-1 (LRP-1), transferrin receptor (TfR), bee venom-derived ion channel, and Aß/LRP-1 related transcytosis complex, respectively. The BBB-permeation efficacies were substantially increased ("boosted") in vector conjugates of P/LLL(40%). We have found that the copolymer group binds at the endothelial membrane and, by an allosterically membrane rearrangement, exposes the sites for vector-receptor complex formation. The specificity of vectors is indicated by competition experiments with nonconjugated vectors. P/LLL(40%) does not function as an inhibitor, suggesting that the copolymer binding site is eliminated after binding of the vector-nanoconjugate. The two-step mechanism, binding to endothelial membrane and allosteric exposure of transcytosis receptors, is supposed to be an integral feature of nanoconjugate-transcytosis pathways. In vivo brain delivery signatures of the nanoconjugates were recapitulated in mouse brains of normal, tumor (glioblastoma), and Alzheimer's disease (AD) models. BBB permeation of the tumor was most efficient, followed by normal and then AD-like brain. In tumor-bearing and normal brains, AP2 was the top performing vector; however, in AD models, D3 and D1 peptides were superior ones. The TfR vector B6 was equally efficient in normal and AD-model brains. Cross-permeation efficacies are manifested through modulated vector coligation and dosage escalation such as supra-linear dose dependence and crossover transcytosis activities.

Urinary Tract Infection in Parkinson's Disease.

Urinary tract infection (UTI) is a common precipitant of acute neurological deterioration in patients with Parkinson's disease (PD) and a leading cause of delirium, functional decline, falls, and hospitalization. Various clinical features of PD including autonomic dysfunction and altered urodynamics, frailty and cognitive impairment, and the need for bladder catheterization contribute to an increased risk of UTI. Sepsis due to UTI is a feared consequence of untreated or undertreated UTI and a leading cause of morbidity in PD. Emerging research suggests that immune-mediated brain injury may underlie the pathogenesis of UTI-induced deterioration of PD symptoms. Existing strategies to prevent UTI in patients with PD include use of topical estrogen, prophylactic supplements, antibiotic bladder irrigation, clean catheterization techniques, and prophylactic oral antibiotics, while bacterial interference and vaccines/immunostimulants directed against common UTI pathogens are potentially emerging strategies that are currently under investigation. Future research is needed to mitigate the deleterious effects of UTI in PD.

Multifunctional Nanopolymers for Blood-Brain Barrier Delivery and Inhibition of Glioblastoma Growth through EGFR/EGFRvIII, c-Myc, and PD-1.

Glioblastoma (GBM) is the most prevalent primary brain cancer in the pediatric and adult population. It is known as an untreatable tumor in urgent need of new therapeutic approaches. The objective of this work was to develop multifunctional nanomedicines to treat GBM in clinical practice using combination therapy for several targets. We developed multifunctional nanopolymers (MNPs) based on a naturally derived biopolymer, poly(ß-L-malic) acid, which are suitable for central nervous system (CNS) treatment. These MNPs contain several anticancer functional moieties with the capacity of crossing the blood-brain barrier (BBB), targeting GBM cells and suppressing two important molecular markers, tyrosine kinase transmembrane receptors EGFR/EGFRvIII and c-Myc nuclear transcription factor. The reproducible syntheses of MNPs where monoclonal antibodies are replaced with AP-2 peptide for effective BBB delivery were presented. The active anticancer inhibitors of mRNA/protein syntheses were Morpholino antisense oligonucleotides (AONs). Two ways of covalent AON-polymer attachments with and without disulfide bonds were explored. These MNPs bearing AONs to EGFR/EGFRvIII and c-Myc, as well as in a combination with the polymer-attached checkpoint inhibitor anti-PD-1 antibody, orchestrated a multi-pronged attack on intracranial mouse GBM to successfully block tumor growth and significantly increase survival of brain tumor-bearing animals.

Interleukin-6 mediates delirium-like phenotypes in a murine model of urinary tract infection.

BACKGROUND: Urinary tract infection (UTI) is frequently implicated as a precipitant of delirium, which refers to an acute confusional state that is associated with high mortality, increased length of stay, and long-term cognitive decline. The pathogenesis of delirium is thought to involve cytokine-mediated neuronal dysfunction of the frontal cortex and hippocampus. We hypothesized that systemic IL-6 inhibition would mitigate delirium-like phenotypes in a mouse model of UTI. METHODS: C57/BL6 mice were randomized to either: (1) non-UTI control, (2) UTI, and (3) UTI + anti-IL-6 antibody. UTI was induced by transurethral inoculation of 1 × 108 Escherichia coli. Frontal cortex and hippocampus-mediated behaviors were evaluated using functional testing and corresponding structural changes were evaluated via quantification of neuronal cleaved caspase-3 (CC3) by immunohistochemistry and western blot. IL-6 in the brain and plasma were evaluated using immunohistochemistry, ELISA, and RT-PCR. RESULTS: Compared to non-UTI control mice, mice with UTI demonstrated significantly greater impairments in frontal and hippocampus-mediated behaviors, specifically increased thigmotaxis in Open Field (p < 0.05) and reduced spontaneous alternations in Y-maze (p < 0.01), while treatment of UTI mice with systemic anti-IL-6 fully reversed these functional impairments. These behavioral impairments correlated with frontal and hippocampal neuronal CC3 changes, with significantly increased frontal and hippocampal CC3 in UTI mice compared to non-UTI controls (p < 0.0001), and full reversal of UTI-induced CC3 neuronal changes following treatment with systemic anti-IL-6 antibody (p < 0.0001). Plasma IL-6 was significantly elevated in UTI mice compared to non-UTI controls (p < 0.01) and there were positive and significant correlations between plasma IL-6 and frontal CC3 (r2 = 0.5087/p = 0.0028) and frontal IL-6 and CC3 (r2 = 0.2653, p < 0.0001). CONCLUSIONS: These data provide evidence for a role for IL-6 in mediating delirium-like phenotypes in a mouse model of UTI. These findings provide pre-clinical justification for clinical investigations of IL-6 inhibitors to treat UTI-induced delirium.

IL-6 Inhibition Reduces Neuronal Injury in a Murine Model of Ventilator-induced Lung Injury.

Mechanical ventilation is a known risk factor for delirium, a cognitive impairment characterized by dysfunction of the frontal cortex and hippocampus. Although IL-6 is upregulated in mechanical ventilation-induced lung injury (VILI) and may contribute to delirium, it is not known whether the inhibition of systemic IL-6 mitigates delirium-relevant neuropathology. To histologically define neuropathological effects of IL-6 inhibition in an experimental VILI model, VILI was simulated in anesthetized adult mice using a 35 cc/kg tidal volume mechanical ventilation model. There were two control groups, as follow: 1) spontaneously breathing or 2) anesthetized and mechanically ventilated with 10 cc/kg tidal volume to distinguish effects of anesthesia from VILI. Two hours before inducing VILI, mice were treated with either anti-IL-6 antibody, anti-IL-6 receptor antibody, or saline. Neuronal injury, stress, and inflammation were assessed using immunohistochemistry. CC3 (cleaved caspase-3), a neuronal apoptosis marker, was significantly increased in the frontal (P < 0.001) and hippocampal (P < 0.0001) brain regions and accompanied by significant increases in c-Fos and heat shock protein-90 in the frontal cortices of VILI mice compared with control mice (P < 0.001). These findings were not related to cerebral hypoxia, and there was no evidence of irreversible neuronal death. Frontal and hippocampal neuronal CC3 were significantly reduced with anti-IL-6 antibody (P < 0.01 and P < 0.0001, respectively) and anti-IL-6 receptor antibody (P < 0.05 and P < 0.0001, respectively) compared with saline VILI mice. In summary, VILI induces potentially reversible neuronal injury and inflammation in the frontal cortex and hippocampus, which is mitigated with systemic IL-6 inhibition. These data suggest a potentially novel neuroprotective role of systemic IL-6 inhibition that justifies further investigation.

Generation of Novel Diagnostic and Therapeutic Exosomes to Detect and Deplete Protumorigenic M2 Macrophages.

Given their protumorigenic function and prevalence in most malignant tumors with lower survival; early detection, and intervention of CD206-positive M2 macrophages may boost the clinical outcome. To determine in vivo distribution of M2 macrophages, 111In-oxine-based radiolabeling of the targeted exosomes is adopted. When these radiolabeled targeted exosomes are injected into breast tumor-bearing mice, exosomes accumulate at the periphery of the primary tumor, metastatic foci in the lungs, spleen, and liver. Ex vivo quantification of radioactivity also shows similar distribution. Injecting DiI dye-labeled exosomes into the same mice shows adherence of exosomes to the CD206-positive M2 macrophages on ex vivo fluorescent microscopy imaging. In addition, these engineered exosomes are utilized to carry the Fc portion of lgG2b with the intention of augmenting antibody-dependent cell-mediated cytotoxicity. It is demonstrated that M2 macrophage targeting therapeutic exosomes deplete M2 macrophages both in vitro and in vivo, and reduce tumor burden, increasing survival in a metastatic breast cancer model.

Influence of Demographic and Reproductive Factors on Cervical Pre-Cancer and Cancer in Bangladesh.

BACKGROUND: In Bangladesh, cervical cancer (CC) is the 2nd most common cancer with estimated 8068 new cases and 5,214 deaths every year. It is also revealed that different socio-demographic factors have association with CC. This study was performed to evaluate the colposcopy outcomes and the association of different demographic and reproductive risk factors with cervical pre-cancer and cancer. METHODS: This retrospective cross-sectional study was carried out at the colposcopy clinic of Bangabandhu Sheikh Mujib Medical University (BSMMU) between January 2010 and December 2016. RESULTS: A total 16147 women attended the colposcopy clinic of BSMMU with VIA positive reports. Among them, 65.73% women were referred from different VIA centers of Dhaka district. Mean age of marriage of the subjects was 16. 93 (± 1) and mean age of 1st delivery was 18.45 years (± 4.10). Almost three-fourth of them were married before 18 years and had their 1st delivery by 20 years. Colposcopy examination of the VIA positive women revealed that 36.7% had CINI, 10.6% had CINII/ III and 7.1% had carcinoma of cervix. Considering CIN as disease the Sensitivity, Specificity, PPV and NPV of colposcopy were found 99.7%, 75.3%, 70.3% and 99.8% respectively. On other hand considering CIN2+ as disease the Sensitivity, Specificity, PPV and NPV of colposcopy were found 73.8%, 92.7%, 64.4% and 95.2% respectively. Statistical analysis revealed that higher age (p=0.000), lower level of education (p=0.007), lower socioeconomic status (p=0.014), higher parity (p=0.001) had individual influence on cervical pre-cancer and cancer. CONCLUSIONS: This study indicated higher age, low level of education, lower socio-economic condition and higher parity as most important socio-demographic factors for developing cervical pre-cancer and cancer in Bangladesh.

Computational Optimization of Bioanalytical Parameters for the Evaluation of the Toxicity of the Phytomarker 1,4 Napthoquinone and its Metabolite 1,2,4-trihydroxynapththalene.

OBJECTIVES: Lawsone (1,4 naphthoquinone) is a non redox cycling compound that can be catalyzed by DT diaphorase (DTD) into 1,2,4-trihydroxynaphthalene (THN), which can generate reactive oxygen species by auto oxidation. The purpose of this study was to evaluate the toxicity of the phytomarker 1,4 naphthoquinone and its metabolite THN by using the molecular docking program AutoDock 4. METHODS: The 3D structure of ligands such as hydrogen peroxide (H2O2), nitric oxide synthase (NOS), catalase (CAT), glutathione (GSH), glutathione reductase (GR), glucose 6-phosphate dehydrogenase (G6PDH) and nicotinamide adenine dinucleotide phosphate hydrogen (NADPH) were drawn using hyperchem drawing tools and minimizing the energy of all pdb files with the help of hyperchem by MM(+) followed by a semi-empirical (PM3) method. The docking process was studied with ligand molecules to identify suitable dockings at protein binding sites through annealing and genetic simulation algorithms. The program auto dock tools (ADT) was released as an extension suite to the python molecular viewer used to prepare proteins and ligands. Grids centered on active sites were obtained with spacings of 54 × 55 × 56, and a grid spacing of 0.503 was calculated. Comparisons of Global and Local Search Methods in Drug Docking were adopted to determine parameters; a maximum number of 250,000 energy evaluations, a maximum number of generations of 27,000, and mutation and crossover rates of 0.02 and 0.8 were used. The number of docking runs was set to 10. RESULTS: Lawsone and THN can be considered to efficiently bind with NOS, CAT, GSH, GR, G6PDH and NADPH, which has been confirmed through hydrogen bond affinity with the respective amino acids. CONCLUSION: Naphthoquinone derivatives of lawsone, which can be metabolized into THN by a catalyst DTD, were examined. Lawsone and THN were found to be identically potent molecules for their affinities for selected proteins.

Role of the HPV DNA test in follow-up of treated cervical intraepithelial neoplasia in Bangladesh.

BACKGROUND: Cervical cancer is a major public health problem in Bangladesh. Persistence of high risk human papillomavirus (HRHPV) influences the progression of the disease, with an important role in follow- up for cervical intraepithelial neoplasia (CIN). OBJECTIVE: To establish application of high risk HPV DNA test in the follow-up of women after treatment of CIN. MATERIALS AND METHODS: This cross-sectional and hospital based study was carried out among 145 CIN treated women during the previous six months to three years at the colposcopy clinic of Bangabandhu Sheikh Mujib Medical University, Dhaka, between January 2011 and June 2012. Pap smear and HPV samples were collected and colposcopy was performed to find out the persistence of the disease. Cervical samples obtained were tested for HPV DNA using the Hybrid Capture II (HC-II) test. A cervical biopsy was collected whenever necessary. The results were compared to assess the efficacy of different methods during follow up such as Pap smear, HPV test and colposcopy. RESULTS: Mean age of the recruited women (n=145) was 33.6 (± 7.6), mean age of marriage was 16.8 (±2.9) and mean age of 1st delivery was 18.8 (±3.5) years. More than half had high grade CIN before treatment and 115 (79.3%) women were managed by LEEP and 20.7% were managed by cold coagulation. Among the 145 treated women, 139 were negative for HPV DNA and six of them (4.1%) were HPV positive. Sensitivity of Pap smear (40.0) and HPV DNA test (40.0) was poor, but specificity was quite satisfactory (>93.0) for all the tests. CONCLUSIONS: The high risk HPV DNA test can be an effective method of identifying residual disease. It can be added to colposcopy and this should be applied to all treated women attending for their first or second post-treatment follow-up visit at 6 months to one year, irrespective of the grade of treated CIN.

Role of print and audiovisual media in cervical cancer prevention in Bangladesh.

BACKGROUND: Visual inspection of cervix with acetic acid (VIA) is offered at 252 centers in 64 districts of Bangladesh. VIA+ve women are managed at colposcopy clinics of Bangabandhu Sheikh Mujib Medical University (BSMMU) and 14 Medical College Hospitals (MCHs). This research work has been supported by 'UICC Cancer Prevention Campaign' programme. OBJECTIVES: This study explored the role of print materials and electronic media to improve cervical cancer screening in the present socio-cultural context of Bangladesh. METHODS: This study was performed from January to August 2011 at two upazilas of Bangladesh (Singair with screening facility and Sonargaon without screening facility). Data were collected by focus group discussion (FGD) with women, husbands and community people before and after intervention. Information on cervical cancer screening and VIA camps was disseminated using advertisement through local cable line of the television, microphone announcement, service providers and leaflet throughout the week prior to a VIA camp. Three-day VIA camps were organized at the upazila health complex (UHC) of both upazilas. Quantitative data was gathered from women at the camps on source of information on VIA and the best method of awareness creation. RESULTS: The population was aware of "cancer" and a notable number knew about cervical cancer. Baseline awareness on prevention and VIA was low and it was negligible where screening services were unavailable. Awareness was increased fourfold in both upazilas after interventions and half of the women and the majority of the community people became aware of screening and available facilities. Cable line advertisement (25.5%), microphone announcement (21.4%), and discussion sessions (20.4%) were effective for awareness creation on VIA. Television was mentioned as the best method (37.4%) of awareness creation. CONCLUSION: Television should be used for nation-wide awareness creation. For local awareness creation, cable line advertisement, microphone announcements and health education at Uthan Baithaks/ EPI sessions can easily be adopted by the government.

Screening for and management of high-grade cervical intraepithelial neoplasia in Bangladesh: a cross-sectional study comparing two protocols.

AIM: To evaluate the feasibility of the 'see and treat' protocol for the management of high-grade cervical intraepithelial neoplasia (CIN) at a colposcopy clinic in Bangladesh. MATERIAL AND METHODS: A cross-sectional and comparative study was carried out between two periods on 358 colposcopy-diagnosed high-grade CIN at the colposcopy clinic of Bangabandhu Sheikh Mujib Medical University (BSMMU). During the first period (January 2005 to June 2008), 181 colposcopy-diagnosed high-grade CIN patients had cervical biopsy by punch biopsy forceps and histology-confirmed high-grade CIN were treated by loop electrosurgical excision procedure (LEEP). During the second period (July 2008 to December 2009), 177 colposcopy-diagnosed high-grade CIN were treated by LEEP at their first visit. RESULTS: During the first and second periods, 48 of 87 and 55 of 73 histology-proven high-grade CIN cases, respectively, received treatment. Among the study population, 37.3% women who had normal or CIN-I in histology were treated unnecessarily in the second period. The compliance of treatment improved by 20% and failure to receive treatment fell by 20%; these changes were statistically significant (P=0.006). CONCLUSION: 'See and treat' protocol is a well-accepted, feasible and useful option for management of high-grade CIN in Bangladesh. It reduces the number of visits to the clinic and failure to receive treatment.

Seroprevalence of Treponema Pallidum antibody among pregnant population of Bangladesh.

AIM: The aim of this study was to investigate the seroprevalence of antibody to Treponema pallidum among pregnant women and its association with age, sexual behavior and socioeconomic status. MATERIALS AND METHODS: In this cross-sectional study, 1266 sera were collected from pregnant women attending Bangabandhu Sheikh Mujib Medical University and Rajshahi, Khulna and Chittagong Medical College Hospitals between February 2007 and April 2008. From each hospital, samples were collected from about a quarter of the total recruited pregnant women. Sera were tested to identify the antibody to T. pallidum using Treponema pallidum hemagglutination at the Virology Department of Bangabandhu Sheikh Mujib Medical University. Logistic regression was used to explore potential risk factors for syphilis. RESULTS: Among 1250 sera tested (16 sera were discarded due to hemolysis), 37 (2.96%) were positive for T. pallidum antibody. Higher seropositivity was associated with couples with no formal education, low socioeconomic status (P = 0.006), marriage at or before 18 years of age (P = 0.002), multiple marriages of the husband (P = 0.031) and sexual activity for more than 10 years (P = 0.007). Among these factors, low education level of women, early age of marriage and multiple marriages of the husband showed individual effects on high seropositivity to T. pallidum. CONCLUSIONS: The high seroprevalence of syphilis among pregnant women attending the antenatal clinics of these hospitals indicates the need for the incorporation of screening for syphilis in routine antenatal screening in Bangladesh. Improvement of education level in the female population, prevention of early marriage and preventing polygamy of husbands may play an important role in prevention of syphilis among women.

Screening for cervical neoplasia in Bangladesh using visual inspection with acetic acid.

OBJECTIVE: To report the organization and early results of a visual inspection with acetic acid (VIA) screening program for cervical cancer prevention in Bangladesh. METHODS: Opportunistic VIA screening was offered by trained medical personnel to healthy women attending healthcare facilities in 44 districts. Colposcopy and/or directed biopsies were performed on VIA-positive women at Bangabandhu Sheikh Mujib Medical University (BSMMU) or medical college hospitals. The numbers of screened and VIA-positive women as well as the results of colposcopy, biopsy, and treatment of those with cervical intraepithelial neoplasia (CIN) attending BSMMU are reported. RESULTS: From January, 2005, to June, 2008, 104098 women underwent VIA screening. Among them, 5013 (4.8%) women were positive, of whom 4371 (87.2%) attended colposcopy clinics in different hospitals. Of the 2188 who underwent colposcopy at BSMMU, 157 were diagnosed with CIN 2-3 and 123 with invasive cancer. For 2065 women (excluding the cancer cases), results for sensitivity, specificity, and positive predictive value of VIA to detect CIN 2-3 lesions were 93.6%, 58.3%, and 15.6%, respectively. Almost half the women with CIN 2 or 3 lesions had treatment. CONCLUSION: A VIA-based program would facilitate the development of screening, diagnosis, and treatment of cervical neoplasia and improve awareness of cervical cancer prevention in Bangladesh.