40 research outputs found
Biogas Production From a Mixture of Cow Manure with Chicken Manure
Biogas technology with zero waste concept is expected to be the alternative energy and to reduce environmental problems. The purpose of this study is to know the biogas yield per kilogram of each chicken and cow manure comparison. The study was conducted in six treatments with the addition of chicken manure of 0, 100, 300, 500, 700 and 1000 grams. The fermentation process is done using a batch system and biogas measurement was taken daily. The parameters to be observed were organic matter, the degree of acidity (pH), temperature, volume of biogas, biogas productivity, and C / N ratio of each treatment. The results showed that the overall pH at the beginning and end of the study tend to be close to neutral. The highest biogas yield was resulted from a mixture of chicken manure and cow manure at the composition of 1:1 or 50%:50% with biogas total amount of 35.690 ml and biogas productivity of 0,33 liters/g (volatile solid)
Proteomics unravels the regulatory mechanisms in human tears following acute renouncement of contact lens use : a comparison between hard and soft lenses
Contact lenses (CLs) provide a superior alternative to spectacles. Although beneficial, the global burden of ocular dysfunctions attributed to regular use of CLs remains a topic of much challenge in ophthalmic research owing to debilitating clinical repercussions on the ocular surface, which are often manifested as breach in tear film integrity. This study elucidated the intricate tear proteome changes attributed to the use of different CLs (hard and soft) and unravelled, for the first time, the restorative mechanisms of several protein clusters following acute renouncement of CL use employing the label-free mass spectrometry-based quantitative proteomics approach. The expression patterns of certain proteins clusters were specific to the use of a particular lens type and a large majority of these actively regulates cell death and survival and, modulates cellular movement on the ocular surface. Noteworthy, CL use also evoked a significant upregulation of glycolytic enzymes associated with hypoxia and corresponding cognate metabolic pathways, particularly glucose metabolism and FXR/RXR pathways. Importantly, the assessment of CL renouncement unravelled the restorative properties of several clusters of proteins involved mainly in organismal injury and abnormalities and, cellular function and maintenance. These proteins play key roles in restoring tear homeostasis and wound-healing mechanisms post-CL use-elicited injury
In vitro anticoagulant activities of Melastoma malabathricum Linn. aqueous leaf extract : a preliminary novel finding.
Limitations of existing anticoagulants have prompted a search for novel agents of natural origin.Fundamentals to this research was the observation that the aqueous leaf extract of Melastoma malabathricum Linn. possesses potent anticoagulant property.In vitro coagulation assays such as
activated partial thromboplastin time (aPTT), prothrombin time (PT) thrombin time (TT) and mixing studies were
performed on citrated plasmas of healthy volunteer donors spiked with different concentrations of the leaf exact
(100-1000μg/ml).The results showed that aPTT of plasma samples spiked with extract was markedly prolonged in
a concentration-dependent manner (p<0.001), but was otherwise for PT and TT. Both types of mixing studies corrected the initially prolonged aPTT to normal range.The extract exhibited no inter-gender variability in its anticoagulant activity.This study highlights that the anticoagulant activity of M. malabathricumaqueous leaf extract affects the intrinsic pathway of the coagulation cascade by causing clotting factor(s) deficiency
Anticoagulant activity and toxicity profiles of Melastoma malabathricum linn. leaf extract
The increased rates of thrombotic diseases contribute to the high number of morbidity
and deaths worldwide each year. For many decades, the use of anticoagulant drugs,
namely warfarin, heparin and their derivatives in the prevention and treatment of these
maladies remain indisputable. Although these anticoagulant agents are efficacious key
players in the clinical practice, they are associated with many well-established
drawbacks that limit their usefulness. Hence, there exists an unmet need for equally
potent but novel anticoagulant agents with improved safety profiles and ease of
administration via the oral route. This study was, thus initiated in view of the current
highlights in the medical realm of anticoagulation and based on the preliminary finding
in a screening study of Melastoma malabathricum Linn. leaf extract. Melastoma leaves
gave the highest extract yield (288.0 ± 1.0 g) when extracted with hot water under reflux,
compared to cold water (143.0 ± 5.5 g) and organic solvent (189.0 ± 0.9 g) extraction methods. Hot water extract was also found to possess potent in vitro anticoagulant
activities, comparable to conventional drug, heparin. Correspondingly, the Melastoma
leaf extract significantly (P < 0.001) prolonged activated partial thromboplastin time
(aPTT) (64 - 300 s) in a concentration-dependent manner (l00-1000 ug/ ml), but did not
affect both prothrombin time (PT) and thrombin time (TT), suggestive of its effect on
the intrinsic pathway of coagulation cascade. There were no inter-gender variations in
the trends of anticoagulant activities. The nature of the anticoagulation caused by the
extract investigated in immediate and timed-incubation mixing studies, demonstrated
that the initially prolonged a PTT of test samples spiked with a range of Melastoma leaf
extract was subsequently corrected to normal clotting time (31.0 - 46.6 s) range when
test samples were subjected to 50 % normal human plasma. Subsequent analysis of
various coagulation factors in the intrinsic pathway showed that Melastoma leaf extract
specifically targeted and caused a considerable deficiency in factor VIII (FVIII) levels
(23 - 35 %) non-dose dependently. Results from the in vivo studies employing suitable
animal models of thrombosis corroborated with the findings of in vitro clot-based assays
and hence, confirmed the inherent anticoagulant properties of Melastoma leaf extract.
The toxic and hemorrhagic propensity of this extract evaluated in an acute oral toxicity
animal study underscored the absence of aberrant effects of the extract in vivo when
administered at a high, single dose (5 g/ kg) for a short period of 14 days. However, subacute
toxicity evaluations of the extract administered on a daily dose in regimen at
various lower dosages (50, 75 and 100 mg/ kg) for 28 days. revealed an array of
abnormal changes, notably hepatic venous dilatation and congestion. and hemorrhage in
renal tissues. Nevertheless, cytotoxicity studies of the extract employing organ-specific
cells and 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) cell viability assay proved the absence of direct toxicity, as substantiated by preserved
cellular integrity and high 50 % inhibitory concentration (ICSO) value of the extract (972
± 2.57 ug/ml), in comparison to evident cytotoxic manifestations of heparin used as
control reference. Collectively, the findings of this study suggested that the hot water
extract of Melastoma leaves have high potential as an affordable alternative for future
development of a potent and safe novel anticoagulant agent of natural origin
Short-Term Omega-3 Supplementation Modulates Novel Neurovascular and Fatty Acid Metabolic Proteome Changes in the Retina and Ophthalmic Artery of Mice with Targeted <i>Cyp2c44</i> Gene Deletion
Cytochrome P450 (CYP) gene mutations are a common predisposition associated with glaucoma. Although the molecular mechanisms are largely unknown, omega-3 polyunsaturated fatty acids (ω-3 PUFA) and their CYP-derived bioactive mediators play crucial roles in the ocular system. Here, we elucidated the proteome and cell-signalling alterations attributed to the main human CYP2C gene deficiency using a homologous murine model (Cyp2c44−/−), and unravelled the effects of acute ω-3 PUFA supplementation in two ocular vascular beds comprising the retrobulbar ophthalmic artery (OA) and retina (R). Male Cyp2c44−/− mice (KO) and their floxed littermates (WT) were gavaged daily for 7 days with 0.01 mL/g of ω-3 PUFA composed of menhaden fish oil. Another group in respective strains served as vehicle-treated controls. OA and R were isolated at day 8 post-treatment (n = 9/group) and subjected to mass spectrometry (MS)-based proteomics and in silico bioinformatics analyses. Cyp2c44−/− resulted in significant detrimental proteome changes associated with compromised vascular integrity and degeneration in the OA and R, respectively. However, notable changes in the OA after ω-3 PUFA intake were associated with the maintenance of intercellular junctional and endothelial cell functions, as well as activation of the fatty acid metabolic pathway in the KO mice. Conversely, ω-3 PUFA supplementation profoundly influenced the regulation of a large majority of retinal proteins involved in the preservation of neuronal and phototransduction activities in WT mice, namely synaptophysin, phosducin and guanylate cyclase-1, while significantly abrogating degenerative processes in the KO mice via the regulation of, namely, synaptotagmin-1 and beta-crystallin B2. In gist, this study demonstrated that dietary supplementation with ω-3 PUFA for a short period of seven days regulated specific neuro-vasculoprotective mechanisms to preserve the functionality of the OA and R in the absence of Cyp2c44. The potential adjunct use of ω-3 PUFA for glaucoma therapy needs further investigation
First insight into the proteome landscape of the porcine short posterior ciliary arteries : key signalling pathways maintaining physiologic functions
Short posterior ciliary arteries (sPCA) provide the major blood supply to the optic nerve head. Emerging evidence has linked structural and functional anomalies of sPCA to the pathogenesis of several ocular disorders that cause varying degrees of visual loss, particularly anterior ischaemic optic neuropathy and glaucoma. Although the functional relevance of this vascular bed is well-recognized, the proteome of sPCA remains uncharacterized. Since the porcine ocular system closely resembles that of the human’s and is increasingly employed in translational ophthalmic research, this study characterized the proteome of porcine sPCA employing the mass spectrometry-based proteomics strategy. A total of 1742 proteins and 10527 peptides were identified in the porcine sPCA. The major biological processes involved in the maintenance of physiological functions of the sPCA included redox and metabolic processes, and cytoskeleton organization. These proteins were further clustered into diverse signalling pathways that regulate vasoactivity of sPCA, namely the tight junction, α- and β-adrenoceptor, 14-3-3, nitric oxide synthase and endothelin-1 -mediated signalling pathways. This study provides the first insight into the complex mechanisms dictating the vast protein repertoire in normal vascular physiology of the porcine sPCA. It is envisioned that our findings will serve as important benchmarks for future studies of sPCA
Effective melanin depigmentation of human and murine ocular tissues: an improved method for paraffin and frozen sections.
PURPOSE: The removal of excessive melanin pigments that obscure ocular tissue morphology is important to address scientific questions and for differential diagnosis of ocular tumours based on histology. Thus, the goal of the present study was to establish an effective and fast melanin bleaching method for paraffin and frozen mouse and human ocular tissues. METHODS: Paraffin-embedded and frozen ocular specimens from mice and human donors were subjected to bleaching employing two methods. The first employed potassium permanganate (KMnO4) with oxalic acid, and the second 10% hydrogen peroxide (H2O2). To determine optimal bleaching conditions, depigmentation was carried out at various incubation times. The effect of diluents used for 10% H2O2 was assessed using phosphate-buffered saline (PBS), and deionized water. Three different slide types and two fixatives, which were ice-cold acetone with 80% methanol, and 4% paraformaldehyde (PFA) were used to determine the optimal conditions for better tissue adherence during bleaching. All tissues were stained in hematoxylin and eosin for histological evaluation. RESULTS: Optimal bleaching was achieved using warm 10% H2O2 diluted in PBS at 65°C for 120 minutes. Chromium-gelatin-coated slides prevented tissue detachment. Adherence of cryosections was also improved with post-fixation using 4% PFA and overnight air-drying at RT after cryosectioning. Tissue morphology was preserved under these conditions. Conversely, tissues bleached in KMnO4/oxalic acid demonstrated poor depigmentation with extensive tissue damage. CONCLUSIONS: Warm dilute H2O2 at 65°C for 120 minutes rapidly and effectively bleached both cryo- and paraffin sections of murine and human ocular tissues
Effective melanin depigmentation of human and murine ocular tissues : an improved method for paraffin and frozen sections
PURPOSE: The removal of excessive melanin pigments that obscure ocular tissue morphology is important to address scientific questions and for differential diagnosis of ocular tumours based on histology. Thus, the goal of the present study was to establish an effective and fast melanin bleaching method for paraffin and frozen mouse and human ocular tissues.
METHODS: Paraffin-embedded and frozen ocular specimens from mice and human donors were subjected to bleaching employing two methods. The first employed potassium permanganate (KMnO4) with oxalic acid, and the second 10% hydrogen peroxide (H2O2). To determine optimal bleaching conditions, depigmentation was carried out at various incubation times. The effect of diluents used for 10% H2O2 was assessed using phosphate-buffered saline (PBS), and deionized water. Three different slide types and two fixatives, which were ice-cold acetone with 80% methanol, and 4% paraformaldehyde (PFA) were used to determine the optimal conditions for better tissue adherence during bleaching. All tissues were stained in hematoxylin and eosin for histological evaluation.
RESULTS: Optimal bleaching was achieved using warm 10% H2O2 diluted in PBS at 65 degrees C for 120 minutes. Chromium-gelatin-coated slides prevented tissue detachment. Adherence of cryosections was also improved with post-fixation using 4% PFA and overnight air-drying at RT after cryosectioning. Tissue morphology was preserved under these conditions. Conversely, tissues bleached in KMnO4/oxalic acid demonstrated poor depigmentation with extensive tissue damage.
CONCLUSIONS: Warm dilute H2O2 at 65°C for 120 minutes rapidly and effectively bleached both cryo- and paraffin sections of murine and human ocular tissues