Konservative versus chirurgische Therapie beim idiopathischen und sekundären Megakolon oder Megarektum im Erwachsenenalter - eine retrospektive multizentrische Kontrollstudie.

BACKGROUND: Idiopathic and secondary megacolon (MC) and megarectum (MR) in adults is associated with persistent bowel dilatation and reduced intestinal motility. Little is known about the optimal treatment of this rare disease. Therefore, we retrospectively analysed long-term data from these patients in 5 community and university hospitals, focusing on conservative versus surgical treatment. METHODS: Patient records from 7/2004 to 9/2021 were screened for colorectal diseases with severe constipation and persistent megacolon ≥ 9.0 cm and/or megarectum ≥ 6.5 cm. Follow up-data was collected through telephone interviews and written surveys. ClinicalTrialsgov NCT04340856. RESULTS: Sixty-seven patients with idiopathic (n=10) and secondary (n=57) MC or MR were identified with only 20 of 67 patients (29.9%) correctly diagnosed. Mean/median age was 64/69 (range 18-93) years. Thirty-two patients were treated with laxative regimens, and 35 underwent surgery (colostomy: n=12, segmental resection, or hemicolectomy: n=10, (sub)total colectomy: n= 13) after conservative treatment attempts in 32/35 (91.4%). The mean/median follow-up was 4.2/2.7 (range 0.1-17.0) years. The readmission rate for MC-associated symptoms was significantly higher after conservative treatment than after surgical therapy at 12 (0.84 vs. 0.36 per patient, p=0.036), 24 (1.00 vs. 0.52, p=0.048) and 36 (1.13 vs. 0.58, p=0.047) months, as was the number of patients with persistent laxative dependence (28/32 (87.5%) vs. 19/33 (57.6%); p = 0.007). Therapy-associated adverse events (Clavien-Dindo classification) were documented more often in surgically treated patients (11/35, 31.4%) (p=0.025). CONCLUSION: Surgical treatment may be considered earlier if idiopathic or secondary MC or MR is correctly diagnosed, and conservative treatment has been attempted.

Influence of online versus traditional learning on EFL listening skills: A blended mode classroom perspective.

Online learning is becoming more and more popular than traditional learning, and the need to investigate its influence within the framework of instruction and learning domains is - in today's emerging, cutting-edge technology world - an academic trend. The current study's problem intends to explore the impact of virtual versus traditional learning on the academic achievement of EFL students, a subject that has garnered substantial attention from English Language Teaching (ELT) researchers. This study aims to clarify if EFL students' academic progress in a listening skills classroom is influenced by traditional or online learning. The present research compiled information on how EFL students performed when English language teaching took place online as opposed to in a traditional classroom environment. This study, using an experimental research design with (N = 30) pairs of students (both male and female), was held at Najran University, KSA, in the academic year 2022-2023 b y using probability (random sampling). This study used pre-and post-tests to gather data from the subjects of the study, bifurcated into controlled and experimental groups employing the two modes of teaching, viz., online and traditional. The findings of the investigation proved that the experimental group achieved better performance compared to the control group in terms of results and scores. There are no significant differences based on gender. In addition, (N = 20) teachers teaching listening skills to EFL learners participated in semi-structured interviews. The qualitative analysis enlisted flexibility, accessibility, effective communication, collaboration, monitoring of student progress, and the use of a blackboard as constructive elements, followed by maintenance costs, wastage of available resources, long-term engagement, limited face-to-face interaction, and demotivation as critical perspectives. According to the findings of this research, the author recommends further studies with more variables.

Ubiquitin ligase CHFR mediated degradation of VE-cadherin through ubiquitylation disrupts endothelial adherens junctions.

Vascular endothelial cadherin (VE-cadherin) expressed at endothelial adherens junctions (AJs) is vital for vascular integrity and endothelial homeostasis. Here we identify the requirement of the ubiquitin E3-ligase CHFR as a key mechanism of ubiquitylation-dependent degradation of VE-cadherin. CHFR was essential for disrupting the endothelium through control of the VE-cadherin protein expression at AJs. We observe augmented expression of VE-cadherin in endothelial cell (EC)-restricted Chfr knockout (ChfrΔEC) mice. We also observe abrogation of LPS-induced degradation of VE-cadherin in ChfrΔEC mice, suggesting the pathophysiological relevance of CHFR in regulating the endothelial junctional barrier in inflammation. Lung endothelial barrier breakdown, inflammatory neutrophil extravasation, and mortality induced by LPS were all suppressed in ChfrΔEC mice. We find that the transcription factor FoxO1 is a key upstream regulator of CHFR expression. These findings demonstrate the requisite role of the endothelial cell-expressed E3-ligase CHFR in regulating the expression of VE-cadherin, and thereby endothelial junctional barrier integrity.

The role of trigger factors in the occurrence of appropriate ICD shocks and their clinical and prognostic implications.

BACKGROUND: The role of triggers in the occurrence of appropriate implantable cardioverter-defibrillator (ICD) shocks due to ventricular tachyarrhythmias is not well known. The aim of the study was to assess the prevalence of trigger factors in appropriate ICD shocks and to analyze their prognostic impact on clinical outcome. METHODS: A total of 710 consecutive patients of a prospective single-center ICD-registry who received a first appropriate ICD shock between 2000 and 9/2021 were analyzed. RESULTS: In 35% of ICD patients with first ICD shock, at least one of the following triggers was found: Ischemia (22%), Compliance (9%), Decompensation (38%), Stress (12%), Technical (5%), Electrolyte/endocrinological disorder (22%) and Medication side effects (4%). The trigger factors can be summarized under the acronym ICD-STEMi. The prospective application of the ICD-STEMi scheme increased the rate of identified triggers from 32% to 56% (p < .001). Patients with triggered first ICD shock had an increased 5-year mortality rate (50% vs. 38%, p < .001). Patients with triggers did not show different mortality outcomes or recurrent ICD shocks whether they received arrhythmia therapy or not. CONCLUSIONS: The evaluation of trigger factors after the occurrence of ICD shocks is mandatory and can be systematically evaluated using the acronym ICD-STEMi. Systematic evaluation of triggers using the ICD-STEMi scheme can identify triggers in about half of ICD patients with first appropriate ICD shock. Patients with triggered ICD shock have a 12% higher 5-year mortality rate.

Prevalence of self-medication and associated factors among female students of health science colleges at Majmaah University: A cross-sectional study.

Introduction: Globally, the prevalence of self-medication among young people has increased exponentially. Due to the basic knowledge and easy access to medicines, undergraduate students at health science colleges are likely to self-medicate. This research was undertaken to assess self-medication prevalence and its contributing factors among female undergraduate students in health science colleges at Majmaah University, Saudi Arabia. Materials and methods: A descriptive, cross-sectional study involving 214 female students from the Majmaah University in Saudi Arabia's health science colleges-Medical: (82, 38.31%) and Applied Medical Science College (132, 61.68%)-was conducted. A self-administered questionnaire with sociodemographic information, drugs used, and reasons for self-medication was used for the survey. Non-probability sampling techniques were used to recruit participants. Results: Of the 214 female participants, 173, 80.84 % (medical: 82, 38.31% and applied medical science: 132, 61.68%) confirmed that they were on self-medication. The majority of participants (42.1%) were between the ages of 20 and 21.5 years (mean ± SD: 20.81 ± 1.4). The main reasons for self-medication were quick relief from the illness (77.5%) followed by saving time (76.3%), minor illnesses (71.1%), self-confidence (56.7%), and laziness (56.7%). The use of leftover drugs at home was common among applied medical science students (39.9%). The main indication for self-medication included menstrual problems (82.7 %), headache (79.8%), fever (72.8%), pain (71.1%), and stress (35.3%). The most common drugs used included antipyretic and analgesics (84.4%), antispasmodics (78.9%), antibiotics (76.9%), antacids (68.2%), multivitamins, and dietary supplements (66.5%). On the contrary, the least used drugs were antidepressants, anxiolytics, and sedatives (3.5, 5.8, and 7.5 %, respectively). Family members were the main source of information for self-medication (67.1%), followed by self-acquired knowledge (64.7%), social media (55.5%), and least were friends (31.2%). For adverse effects of the medication, the majority of them consulted the physician (85%) followed by consulting the pharmacist (56.7%) and switched to other drugs or decreased drug dosage. Quick relief, saving time, and minor illness were the main reasons for self-medication among health science college students. It is recommended to conduct awareness programs, workshops, and seminars to educate on the benefits and adverse effects of self-medication.

Chemical profile, anti 5-lipoxygenase and cyclooxegenase inhibitory effects of ginger (Zingiber officinale) rhizome, callus and callus treated with elicitors / Perfil químico, anti-5-lipoxigenase e efeitos inibitórios da ciclooxegenase do rizoma de gengibre (Zingiber officinale), do calo e calos tratados com elicitores

The present study investigated the chemical profiles and evaluated the inhibitory effect against 5-Lipoxygenase (5-Lox) activity for extracts of ginger rhizome, callus, and callus treated with the elicitors; yeast extract (100, 300 and 500 mg/L), glycine (100, 200 and 300 mg/L) and salicylic acid (100 and 200 mg/L). Oils and chloroform: methanol (CM) extracts were prepared by maceration in petroleum ether and CM (1:1, v/v), respectively. Chemical profiles were determined by gas chromatography/mass spectrometry (GC/MS) analysis. Oil of the callus recorded higher 5-Lox inhibitory effect (IC50 58.33±4.66 µg/mL) than the oil of rhizome (IC50168.34±15.64 µg/mL) and comparable to that of the positive control; Nordihydroguaiaretic acid (IC50 61.25±1.02 µg/mL). The chemical profile of the callus oil contained large amounts of fatty acids, mainly the unsaturated fatty acid oleic acid (31.11%) and saturated fatty acid palmitic acid (28.56%). Elicitors modified the chemical profile of the callus and ameliorated the anti-5-Lox activity of CM extract of the callus. CM extracts of callus treated with 100 and 300 mg/L yeast extract and 50 mg/L salicylic acid significantly suppressed (P ≤ 0.05) the 5-Lox activity by 33.16%, 25.46% and 16%, respectively as compared to the CM extract of untreated callus. In conclusion, ginger callus could be considered as a valuable dietary supplement in the treatment of various inflammatory disorders.

O presente estudo teve como objetivo investigar os perfis químicos e avaliar o efeito inibitório da atividade da 5-Lipoxigenase (5-Lox) em extratos de rizoma, calo e calo de gengibre tratados com os eliciadores; extrato de levedura (100, 300 e 500 mg / L), glicina (100, 200 e 300 mg / L) e ácido salicílico (100 e 200 mg / L). Extratos de óleos e clorofórmio: metanol (CM) foram preparados por maceração em éter e CM (1: 1, v / v), respectivamente. Os perfis químicos foram determinados por análise de cromatografia gasosa / espectrometria de massa (GC / MS). O óleo do calo registrou maior efeito inibitório de 5-Lox (IC50 58,33 ± 4,66 µg / mL) do que o óleo de rizoma (IC50168,34 ± 15,64 µg / mL) e comparável ao do controle positivo; Ácido nordi-hidroguaiarético (IC50 61,25 ± 1,02 µg / mL). O perfil químico do óleo de calo continha grandes quantidades de ácidos graxos, principalmente o ácido graxo insaturado ácido oleico (31,11%) e ácido graxo saturado palmítico (28,56%). Os elicitores modificaram o perfil químico do calo e melhoraram a atividade anti-5-Lox do extrato de CM do calo. Extratos de CM de calos tratados com 100 e 300 mg / L de extrato de levedura e 50 mg / L de ácido salicílico suprimiram significativamente (P ≤ 0,05) a atividade de 5-Lox em 33,16%, 25,46% e 16%, respectivamente, em comparação com o extrato de CM de calo não tratado. Em conclusão, o calo de gengibre pode ser considerado um suplemento dietético valioso no tratamento de vários distúrbios inflamatórios.

Structural equation modelling analysis determining causal role among methyltransferases, methylation, and apoptosis during human pregnancy and abortion.

The human implantation failure during first trimester leads to spontaneous abortions. Spontaneous abortions are consecutive and occur twice or thrice (with or without prior live births) due to factors which are either maternal or fetal. However, it also constitutes of unknown etiology; known as unexplained recurrent spontaneous abortions (URSA). In this study, the medical terminated human normal early pregnancies (NEP) of the first trimester were taken as control samples, the normal decidual sample whose molecular and epigenetic changes were compared with that of decidua of human URSA subjects. Apoptosis-related genes reported in consecutive recurrent pregnancy loss became the basis for this study. So, in this study, we evaluated the hypothesis that "p53 methylation level through methyltransferases (G9aMT and DNMT1) implicates the fate of embryo towards sustenance or cessation of pregnancy". Further, the interaction between P53, BAX, BCL-2, CASPASE-6, G9aMT, DNMT-1, and methylated p53 expression level(s) during the first trimester of both URSA and NEP are included in this study. The degree of p53 methylation during the first trimester is found to be significant and positively correlated with that of G9aMT (p < 0.05), BCL-2 (p < 0.001), and DNMT1 (p < 0.001) at both transcript and protein level. A significant and negative correlation (with p-value < 0.001) between the degree of p53 methylation during the first trimester and that of the expression level of TUNEL assay (Apoptosis), P53, BAX, and CASPASE-6 are also observed in the present study. A positive correlation between apoptosis and a higher level of p53 expression (which is possibly due to low degree of p53 methylation) is observed both at the transcript and protein level in URSA which is in line with our findings. The analysis performed using structural equation modelling (SEM) further throws light on the causal relationship between sustenance of pregnancy or URSA during the first trimester of a human pregnancy and degree of methylation of p53 which is closely correlated with the interaction between G9aMT, DNMT1, BCL-2, BAX, P53, CASPASE-6, and apoptosis.

Bio-mediated synthesis of 5-FU based nanoparticles employing orange fruit juice: a novel drug delivery system to treat skin fibrosarcoma in model animals.

Nano-sized drug delivery systems (NDDS) have been widely exploited to achieve targeted delivery of pharmaco-materials. Traditional pharmaceutical approaches, implied in the synthesis of nano-formulations, are obscure owing to the incompatible physico-chemical properties of the core drug as well as some other factors crucial in development of NDDS. Infact, most of the existing methods used in development of NDDS rely on usage of additives or excipients, a special class of chemicals. Barring few exceptions, the usage of synthetic excipients ought to be curtailed because of several associated undesirable features. Such issues necessitate strategies that lead to development of the synthetic excipient free drug delivery system. Plant based extracts have great potential to induce synthesis of nano-sized particles. Considering this fact, here we propose a prototype employing orange fruit juice (OJ) to facilitate bio-mediated synthesis of nano-sized supra-molecular assemblies of 5-fluorouracil (5-FU), a potent anticancer drug. The as-synthesized 5-FU Nanoparticles (NPs) retained the anti-neoplastic efficacy of the parent compound and induced apoptosis in cancer cells. The novel 5-FU NPs formulation demonstrated enhanced efficacy against DMBA induced experimental fibrosarcoma in the mouse model when compared to the micro-sized crystals of parent 5-FU drug.

Resident-Level Factors Associated with Hospitalization Rates for Newly Admitted Long-Term Care Residents in Canada: A Retrospective Cohort Study.

Chez les résidents en soins de longue durée (SLD), l'hospitalisation peut amener des complications telles que le déclin fonctionnel. L'objectif de notre étude était d'examiner l'association entre les données démographiques et de santé et le taux d'hospitalisation des résidents nouvellement admis en SLD. Nous avons mené une étude de cohorte rétrospective incluant tous les centres de SLD de six provinces et d'un territoire du Canada, à l'aide des données de la RAI-MDS 2.0 et de la Discharge Abstract Database. Nous avons inclus les résidents nouvellement admis ayant eu une évaluation entre le 1er janvier et le 31 décembre 2013 (n = 37 998). Les résidents de sexe masculin avec une santé plus instable et une déficience fonctionnelle de modérée à grave présentaient des taux d'hospitalisation plus élevés, tandis que les résidents avec une déficience cognitive de modérée à grave avaient des taux moindres. Les résultats de notre étude pourraient contribuer à l'identification des résidents nouvellement admis qui seraient plus à risque d'hospitalisation et à l'élaboration de stratégies préventives plus ciblées, incluant la réadaptation, la planification préalable de soins, les soins palliatifs et les services gériatriques spécialisés.Hospitalizations of long-term care (LTC) residents can result in adverse outcomes such as functional decline. The objective of our study was to investigate the association between demographic and health information and hospitalization rate for newly admitted LTC residents. We conducted a retrospective cohort study of all LTC homes in six provinces and one territory in Canada, using data from the Resident Assessment Instrument­Minimum Data Set (RAI-MDS) 2.0 and the Discharge Abstract Database. We included newly admitted residents with an assessment between January 1 and December 31, 2013 (n = 37,998). Residents who were male, had higher health instability, and had moderate or severe functional impairment had higher rates of hospitalization, whereas residents who had moderate or severe cognitive impairment had decreased rates. The results of our study can be used to identify newly admitted residents who may be at risk for hospitalization, and appropriately target preventative interventions, including rehabilitation, advance care planning, palliative care, and geriatric specialty services.

Phytochemicals as modulators of M1-M2 macrophages in inflammation.

Macrophages are critical mediators of the innate immune response against foreign pathogens, including bacteria, physical stress, and injury. Therefore, these cells play a key role in the "inflammatory pathway" which in turn can lead to an array of diseases and disorders such as autoimmune neuropathies and myocarditis, inflammatory bowel disease, atherosclerosis, sepsis, arthritis, diabetes, and angiogenesis. Recently, more studies have focused on the macrophages inflammatory diseases since the discovery of the two subtypes of macrophages, which are differentiated on the basis of their phenotype and distinct gene expression pattern. Of these, M1 macrophages are pro-inflammatory and responsible for inflammatory signaling, while M2 are anti-inflammatory macrophages that participate in the resolution of the inflammatory process, M2 macrophages produce anti-inflammatory cytokines, thereby contributing to tissue healing. Many studies have shown the role of these two subtypes in the inflammatory pathway, and their emergence appears to decide the fate of inflammatory signaling and disease progression. As a next step in directing the pro-inflammatory response toward the anti-inflammatory type after an insult by a foreign pathogen (e. g., bacterial lipopolysaccharide), investigators have identified many natural compounds that have the potential to modulate M1 to M2 macrophages. In this review, we provide a focused discussion of advances in the identification of natural therapeutic molecules with anti-inflammatory properties that modulate the phenotype of macrophages from M1 to M2.

Biomimetically engineered Amphotericin B nano-aggregates circumvent toxicity constraints and treat systemic fungal infection in experimental animals.

Biomimetic synthesis of nanoparticles offers a convenient and bio friendly approach to fabricate complex structures with sub-nanometer precision from simple precursor components. In the present study, we have synthesized nanoparticles of Amphotericin B (AmB), a potent antifungal agent, using Aloe vera leaf extract. The synthesis of AmB nano-assemblies (AmB-NAs) was established employing spectro-photometric and electron microscopic studies, while their crystalline nature was established by X-ray diffraction. AmB-nano-formulation showed much higher stability in both phosphate buffer saline and serum and exhibit sustained release of parent drug over an extended time period. The as-synthesized AmB-NA possessed significantly less haemolysis as well as nephrotoxicity in the host at par with Ambisome®, a liposomized AmB formulation. Interestingly, the AmB-NAs were more effective in killing various fungal pathogens including Candida spp. and evoked less drug related toxic manifestations in the host as compared to free form of the drug. The data of the present study suggest that biomimetically synthesized AmB-NA circumvent toxicity issues and offer a promising approach to eliminate systemic fungal infections in Balb/C mice.

TLR Agonist Augments Prophylactic Potential of Acid Inducible Antigen Rv3203 against Mycobacterium tuberculosis H37Rv in Experimental Animals.

In general, the members of Lip gene family of Mycobacterium tuberculosis evoke strong immune response in the host. Keeping this fact into consideration, we investigated role of Rv3203, a cell wall associated protein with lipolytic activity, in imparting protection against experimental murine tuberculosis. The data of the present study suggested that archaeosome encapsulated Rv3203 induce strong lymphocyte proliferation, up-regulated Th-1 biased cytokines profile, increased expression of co-stimulatory markers on both antigen presenting cells and T lymphocytes. The immuno-prophylactic response was further modulated by exposure of the animals to zymosan, a TLR2/6 agonist, prior to immunization with archaeosome encapsulated Rv3203. Interestingly, pre-treatment of experimental animals with zymosan boosted strong immunological memory as compared to archaeosome encapsulated Rv3203 as well as BCG vaccine. We conclude that priming of immunized animal with TLR agonist followed by immunization with archaeosomes encapsulated Rv3203 offer substantial protection against tuberculosis infection and could be a potential subunit vaccine based prophylactic strategy.

Pharmacophore hybrid approach of new modulated bis-diimine Cu(II)/Zn(II) complexes based on 5-chloro Isatin Schiff base derivatives: Synthesis, spectral studies and comparative biological assessment.

Novel bioactive 5-chloro isatin based Schiff base ligands, (N,N'E,N,N'Z)-N,N'-(5-chloroindoline-2,3-diylidene)bis(5-nitrobenzo [d]thiazol-2-amine), L(1) and (N,N'E,N,N'Z)-N,N'-(5-chloroindoline-2,3-diylidene)bis(5-nitrothiazol-2-amine), L(2) derived from 2-amino 5-nitrobenzothiazole and 2-amino 5-nitrothiazole and their metal complexes, [Cu(L(1))2]Cl2;1, [Zn(L(1))2(H2O)2]Cl2;2, [Cu(L(2))2]Cl2;3 and [Zn(L(2))2(H2O)2]Cl2;4 have been synthesized. The composition, stoichiometry and geometry of the proposed ligands and their complexes have been envisaged by the results of elemental analyses and spectroscopic data (FT-IR, (1)H NMR and (13)C NMR, Mass and EPR). The molar conductivity values of the metal complexes revealed their ionic nature. The thermal stability of metal complexes was demonstrated by TGA/DTA studies while the crystalline nature of the complexes has been ascertained by XRD. Furthermore, a comparative account of in vitro antibacterial study against different bacterial strains with respect to standard antibiotic and scavenging activity against standard control at different concenterations unfolded pronounced antibacterial and radical scavenging potencies of the metal complexes as compared to free ligands. In addition, in vitro cytotoxicity of ligands and its metal complexes was also screened on MCF7 (Human breast adenocarcinoma), HeLa (Human cervical carcinoma) and HepG2 (Human Hepatocellular carcinoma), cell lines and normal cells (PBMC). The antiproliferative outcomes revealed that metal complexes exhibit superior activity in general as compared to free ligands (L(1) and L(2)) where metal complexes (1 and 2) of 5-chloro isatin linked benzothiazole motif (L(1)) are found to have better prospect of acting as chemotherapeutic agents which can be explained in terms of greater biopotency, planarity and conjugation against all the tested cancer cell lines with IC50<2.80 µM.

Synthesis and characterization of Schiff base octaazamacrocyclic complexes and their biological studies.

A condensation reaction between 1,2-diphenylethane-1,2-dione dihydrazone (DPEDDH) and dimethyl or diethyloxalate in methanol resulted in a novel Schiff base octaazamacrocyclic ligand, (L): (6,7,14,15-tetraoxa-2,3,10,11-tetraphenyl-1,4,5,8,9,12,13,16-octaazacyclohexadecane-1,3,9,11-tetraene). Subsequently metal complexes of the type [MLX2] and [CuL]X2; (M=Mn(II), Co(II), Ni(II) and Zn(II); X=Cl or NO3) were synthesized by the reaction of the free macrocyclic ligand (L) with the corresponding metal salts in 1:1 molar ratio. These complexes were characterized on the basis of analytical data, molar conductivity and magnetic susceptibility measurements, ESI-mass, IR, NMR ((1)H and (13)C), EPR and electronic spectral studies. The thermal stability of the complexes was also studied by TGA and DTA analyses. These studies show that all the complexes have octahedral arrangement around the metal ions except copper complexes which are square planar. The ligand and its complexes were screened for their antibacterial activity in vitro against Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacteria and were also studied for their anticancer activity against the human cancer cells lines: HeLa (Human cervical carcinoma), MCF7 (Human breast adenocarcinoma) and Hep3B (Human Hepatocellular carcinoma). The recorded IC50 values for the tested compounds show moderate to good cytotoxicity against these cancer cell lines. The copper complex, [CuL]Cl2, showed excellent antimicrobial activity against tested microorganisms which is almost equivalent to the standard drug ciprofloxacin.

Escheriosome-mediated cytosolic delivery of PLK1-specific siRNA: potential in treatment of liver cancer in BALB/c mice.

AIM: In the present study, the anticancer efficacy of a novel escheriosome-based formulation of PLK1-specific siRNA was evaluated against liver cancer in BALB/c mice. MATERIALS & METHODS: The escheriosome-based siRNA nanoparticles were prepared using lipids isolated from Escherichia coli. The escheriosomes were characterized for size, surface charge and stability. The anticancer potential of PLK1-specific siRNA formulation was ascertained on the basis of expression of pro-/anti-apoptotic factors and histopathological studies. RESULTS: The escheriosome-entrapped siRNA was found to be released in surrounding milieu in a sustained manner. The nanoformulation was successful in modulating proapoptotic factors and eventually helped in better survival of the treated animals. CONCLUSION: Our data demonstrate the efficacy of systemically administered siRNA in the treatment of experimental liver cancer. This novel therapeutic strategy may be applicable to a broad range of cancers in patients with the obstinate form of the disease.

Development, characterization and efficacy of niosomal diallyl disulfide in treatment of disseminated murine candidiasis.

In the current study, a novel niosome based formulation of diallyl disulfide (DADS) was evaluated for its potential to treat disseminated candidiasis in mouse model. Among various non-ionic surfactants tested, niosome formulation prepared using Span 80 was found to be most efficient in the entrapment of DADS. The DADS loaded niosomes had size dimensions in the range of 140 ± 30 nm with zeta potential of -30.67 ± 4.5. Liver/kidney function tests as well as histopathologic studies suggested that noisome-based DADS formulations are safe at the dose investigated. When administered to Candida albicans infected animals, the DADS bearing niosomal formulation cleared the fungal burden and increased their survival much efficiently than its free form. FROM THE CLINICAL EDITOR: In this study, a novel niosomal formulation of the antifungal DADS was utilized in a murine candidiasis model, resulting in more efficient fungal clearance compared to the free formulation.

Tissue plasminogen activator and plasminogen activator inhibitor-1 levels in patients with acute myocardial infarction and unstable angina.

OBJECTIVE: To compare levels of tissue plasminogen activator and plasminogen activator inhibitor-1 levels in patients with acute myocardial infarction and unstable angina in order to understand the use of high sensitivity C-reactive protein (hsCRP), coagulation and fibrinolysis markers for cardiovascular risk assessment. METHODS: The cross-sectional case-control study compared circulating concentrations of high sensitivity C reactive protein (hsCRP), fibrinogen, tissue-type plasminogen activator (tPA), and plasminogen activator inhibitor-1 (PAI-1) between patients of acute myocardial infarction (AMI) (n = 67), Unstable Angina Pectoris (UA) (n = 35) and healthy control subjects (n = 39) at the King Khalid University Hospital, Riyadh, Saudi Arabia, from June 2006 to August 2007. RESULTS: The patients had significantly higher hsCRP (1.06 +/- 0.11 vs 0.52 +/- 0.14, p < 0.01), fibrinogen (426.21 +/- 24.09 vs 329.32 +/- 13.93, p < 0.05), PAL-1 (44.02 +/- 6.05 vs 19.35 +/- 3.94, p < 0.01) and tPA (12.31 +/- 1.16 vs 9.49 +/- 0.86, p < 0.05) compared to the controls. Fibrinogen (329.32 +/- 13.93) and PAL-1 (19.35 +/- 3.94) were higher in both angina and infarction groups compared to the healthy subjects (p < 0.01). Between the two categories of patients the difference between Fibrinogen (449.60 +/- 52.98 vs 419.46 +/- 23.42) and PAL-1 (52.00 +/- 17.34 vs 43.19 +/- 6.10) levels were non-significant. Also, the difference in tPA levels between the controls and angina patients was nonsignificant (9.49 +/- 0.86 vs 9.91 +/- 1.24 p > 0.05). It was higher in infarction patients (14.79 +/- 3.14) compared to angina patients and the controls, (p < 0.05). Compared to the controls, hsCRP levels were significantly higher in both the patient groups (0.52 +/- 0.14, 1.05 +/- 0.28, 1.40 +/- 0.20, p < 0.01). Moreover, they were significantly higher in infarction patients than those suffering from angina (p < 0.05). CONCLUSIONS: CAD patients had a procoagulant state and presented with higher levels of hsCRP compared to the healthy individuals. Moreover, there were significant differences in coagulation markers and hsCRP between angina and infarction patients.

Characterization of a thermostable lipase showing loss of secondary structure at ambient temperature.

A gene encoding extracellular lipase was cloned and characterized from metagenomic DNA extracted from hot spring soil. The recombinant gene was expressed in E. coli and expressed protein was purified to homogeneity using hydrophobic interactions chromatography. The mature polypeptide consists of 388 amino acids with apparent molecular weight of 43 kDa. The enzyme displayed maximum activity at 50 °C and pH 9.0. It showed thermal stability up to 40 °C without any loss of enzyme activity. Nearly 80% enzyme activity was retained at 50 °C even after incubation for 75 min. However above 50 °C the enzyme displayed thermal instability. The half life of the enzyme was determined to be 5 min at 60 °C. Interestingly the CD spectroscopic study carried out in the temperature range of 25-95 °C revealed distortion in solution structure above 35 °C. However the intrinsic tryptophan fluorescence spectroscopic study revealed that even with the loss of secondary structure at 35 °C and above the tertiary structure was retained. With p-nitrophenyl laurate as a substrate, the enzyme exhibited a K ( m ), V ( max ) and K ( cat ) of 0.73 ± 0.18 µM, 239 ± 16 µmol/ml/min and 569 s(-1) respectively. Enzyme activity was strongly inhibited by CuCl(2), HgCl(2) and DEPC but not by PMSF, eserine and SDS. The protein retained significant activity (~70%) with Triton X-100. The enzyme displayed 100% activity in presence of 30% n-Hexane and acetone.

Engineering of a metagenome derived lipase toward thermal tolerance: effect of asparagine to lysine mutation on the protein surface.

A highly thermostable mutant lipase was generated and characterized. Mutant enzyme demonstrated 144 fold enhanced thermostability over the wild type enzyme at 60°C. Interestingly, the overall catalytic efficiency (k(cat/)K(m)) of mutant was also enhanced (~20 folds). Circular dichroism spectroscopy, studied as function of temperature, demonstrated that the mutant lipase retained its secondary structure up to 70-80°C, whereas wild type protein structure was completely distorted above 35°C. Additionally, the intrinsic tryptophan fluorescence (a probe for the tertiary structure) also displayed difference in the conformation of two enzymes during temperature dependent unfolding. Furthermore, mutation N355K resulted in extensive H-bonding (Lys355 HZ1OE2 Glu284) with a distance 2.44 Å. In contrast to this, Wt enzyme has not shown such H-bonding interaction.

Escheriosome mediated cytosolic delivery of Candida albicans cytosolic proteins induces enhanced cytotoxic T lymphocyte response and protective immunity.

Protection against intracellular fungal infections, in a manner similar to viral challenges necessitates activation of both humoral and cell mediated immune responses in unison. Most of the presently available antigen delivery vehicles including egg phosphatidyl-choline (egg-PC) liposomes, can evoke mainly humoral immune responses in the immunized animals. Keeping this fact into consideration, we earlier developed Escherichia coli membrane lipid vesicles (escheriosomes) and demonstrated that escheriosomes successfully fuse with the plasma membrane of macrophages ensuing in effective cytoplasmic delivery of entrapped antigen, a pre-requisite for inducing CD8(+) T cell response against antigens. In the present study, we report the ability of escheriosomes encapsulated Candida albicans (C. albicans) cytosolic antigens (cAg), to generate protective immunity against systemic C. albicans infection in BALB/c mice. The immunization schedule using escheriosome encapsulated cAg induced strong antigen-specific CD8(+) T-cell responses, which were markedly higher than that observed in mice immunized with IFA-antigen emulsion, or antigen encapsulated in egg PC liposomes. Interestingly, immunization with cAg delivered in escheriosomes was also successful in complete elimination of C. albicans infection in Balb/c mice. The study suggests that escheriosomes may function as a novel immunoadjuvants and emerge as an effective tool for generating protective immunity against C. albicans infection.