Phytochemical Evaluation, Antioxidant and Antimicrobial Activities of Various Extracts from Leaves and Stems of Bryophyllum pinnatum

Antioxidant and antimicrobial activities of different extracts (methanol and ethyl acetate) of leaf and stem of Bryophyllum pinnatum were studied. The screening for the secondary metabolites was carried out using the standard methods. The antioxidant capacities of the different extracts were assessed using DPPH (2,2-diphenyl-1-picrylhydrazyl) radicals and Ferric reducing antioxidant power (FRAP) while the antimicrobial activity of the extracts obtained were screened against Gram-positive, Gram-negative bacteria and fungi (Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa Salmonella spp., Vibrio cholerae, Candida albicans and Aspergillus niger) using Agar well diffusion method. Both extracts obtained from leaf and stem of Bryophyllum pinnatum contained most of the phytochemical compounds tested for. However, anthocyanins and anthraquinone were not detected in leaf extracts while coumarin was absent in stem extracts. Quantification of bioactive compounds showed that both extracts contained the highest concentration of polyphenols (34.49 ±0.47 mg GAE/g and 32.32 ±1.2 mg GAE/g for methanol leaf and stem extracts respectively) while the least concentration was recorded for alkaloids (0.03±0.02 mg/g for methanol stem extract). Results revealed that the extracts showed dose-dependent scavenging of DPPH as well as the ability of the extracts to reduce FeCl3 solution, with methanol extracts exhibiting the highest scavenging and reducing capacity. However the leaves of Bryophyllum pinnatum had greater antioxidant activity than the stem by DPPH and ferric reducing assays, with IC50 values ranging from 3.147µg/ml to 3.80µg/ml for DPPH and 331.9 - 451 µg/ml for FRAP assays. The antimicrobial activity of various solvent extracts of leaf and stem reveal that microorganisms exhibited different sensitivities towards these extracts in a dose-dependent manner. Methanol leaf extract showed no activity against Escherichia coli while Pseudomonas aeruginosa was insensitive to ethyl acetate leaf extract. For stem extracts, Aspergillus niger, Vibrio cholerae and Pseudomonas aeruginosa were resistant to methanol extract while Aspergillus niger, Salmonella spp. and Pseudomonas aeruginosa was resistant towards ethyl acetate stem extract. The results obtained in this study showed that Bryophyllum pinnatum is a reservoir of bioactive compounds and both extracts exhibited significant antimicrobial and antioxidant activity

Usefulness of the lupus low disease activity state as a treatment target in childhood-onset SLE

Objective Treat-to-target (T2T) strategies are advocated to improve prognosis in childhood-onset SLE (cSLE). Proposed T2T states include SLEDAI score of <4 (SLEDAI-LD), limited corticosteroid use (low-CS), and lupus low disease activity state (LLDAS). We sought to compare T2T states for their association with cSLE prognosis under consideration of relevant disease characteristics such as pre-existing damage, race and lupus nephritis (LN).Methods Longitudinal data from 165 patients enrolled in the Cincinnati Lupus Registry were included. LN presence was based on renal biopsy, and patients were followed up until 18 years of age.Results The 165 patients (LN: 45, white: 95) entered the registry within a median of 0 (IQR: 0–1) year post diagnosis and were followed up for a median of 4 (IQR: 2–5) years during which 80%, 92% and 94% achieved LLDAS, low-CS and SLEDAI-LD. Patients with LN were significantly less likely to achieve any T2T state (all p<0.03) and required a significantly longer time to reach them (all p<0.0001). Over the study period, patients maintained low-CS, SLEDAI-LD or LLDAS for a median of 76% (IQR: 48%–100%), 86% (IQR: 55%–100%) or 39% (IQR: 13%–64%) of their follow-up. Significant predictors of failure to maintain LLDAS included LN (p≤0.0062), pre-existing damage (p≤0.0271) and non-white race (p≤0.0013). There were 22%, 20% and 13% of patients who reached SLEDAI-LD, CS-low and LLDAS and nonetheless acquired new damage. Patients with LN had a higher risk of new damage than patients without LN even if achieving low-CS (p=0.009) or LLDAS (p=0.04).Conclusions Patients with LN and pre-existing damage are at higher risk of increased future damage acquisition, even if achieving a T2T state such as LLDAS. Among proposed common T2T states, the LLDAS is the hardest to achieve and maintain. The LLDAS may be considered the preferred T2T measure as it conveys the highest protection from acquiring additional disease damage

The Transcription Factor C/EBP-β Mediates Constitutive and LPS-Inducible Transcription of Murine SerpinB2

SerpinB2 or plasminogen activator inhibitor type 2 (PAI-2) is highly induced in macrophages in response to inflammatory stimuli and is linked to the modulation of innate immunity, macrophage survival, and inhibition of plasminogen activators. Lipopolysaccharide (LPS), a potent bacterial endotoxin, can induce SerpinB2 expression via the toll-like receptor 4 (TLR4) by ~1000-fold over a period of 24 hrs in murine macrophages. To map the LPS-regulated SerpinB2 promoter regions, we transfected reporter constructs driven by the ~5 kb 5'-flanking region of the murine SerpinB2 gene and several deletion mutants into murine macrophages. In addition, we compared the DNA sequence of the murine 5′ flanking sequence with the sequence of the human gene for homologous functional regulatory elements and identified several regulatory cis-acting elements in the human SERPINB2 promoter conserved in the mouse. Mutation analyses revealed that a CCAAT enhancer binding (C/EBP) element, a cyclic AMP response element (CRE) and two activator protein 1 (AP-1) response elements in the murine SerpinB2 proximal promoter are essential for optimal LPS-inducibility. Electrophoretic mobility shift (EMSA) and chromatin immunoprecipitation (ChIP) assays demonstrated that LPS induces the formation of C/EBP-β containing complexes with the SerpinB2 promoter. Importantly, both constitutive and LPS-induced SerpinB2 expression was severely abrogated in C/EBP-β-null mouse embryonic fibroblasts (MEFs) and primary C/EBP-β-deficient peritoneal macrophages. Together, these data provide new insight into C/EBP-β-dependent regulation of inflammation-associated SerpinB2 expression