Descelularización y regeneración de tráquea porcina: un acercamiento a la ingeniería tisular / Decellularization and Regeneration of Porcine Trachea: an Approach to Tissue Engineering

Resumen Antecedentes: la ingeniería tisular permite obtener órganos como injertos a partir de tejidos descelularizados, regenerados con células autólogas. Objetivo: descelularizar y regenerar tráqueas porcinas. Material y métodos: se descelularizaron tráqueas porcinas colocándolas cada una en el epiplón de cuatro cerdos Yorkshire para su regeneración in vivo. Una tráquea desce-lularizada con tritón (DT), descelularizada con desoxicolato (DD), descelularizada con desoxicolato y reforzada con un polímero y células epiteliales (DDR), y una nativa crio-preservada (NC). Después de 8 días se obtuvieron la DD, NC y DDR; y al día 15, la DT. Se las evaluó mecánica e histológicamente, se realizó el análisis casuístico. Resultados: las tráqueas descelularizadas conservaron la integridad del cartílago, sin diferencias mecánicas, excepto la DDR con mayor rigidez. Las tráqueas regeneradas presentaron menor rigidez, excepto la DDR que además perdió el epitelio y la vascula-ridad. Las DT, DD mostraron epitelio no respiratorio, fibrosis y vasculogénesis con in-flamación. Conclusiones: las matrices conservaron sus características mecánicas. La regenera-ción in vivo ofrece ventajas como la esterilidad, interacción celular, nutrientes; es senci-llo, factible y económico, pero no hay control del crecimiento celular y vascularización, y los tejidos presentaron alteraciones mecánicas e histológicas. El polímero impidió la re-epitelialización y revascularización. Este estudio abre la posibilidad de mejorar las me-todologías de ingeniería tisular aplicadas al tejido traqueal.

Abstract Introduction: tissue engineering makes it possible to obtain organs as grafts from de-cellularized tissues, regenerated with autologous cells.Objective: decellularize and regenerate porcine tracheas.ARTÍCULO ORIGINAL | Respirar, 2023; 15(3): 188-199 | ISSN 2953-3414 | https://doi.org/10.55720/respirar.15.3.5RECIBIDO: 9 agosto 2023ACEP TADO: 31 agosto 2023 Elisa Barrera-Ramírezhttps://orcid.org/0000-0002-2778-0882Rubén Efraín Garrido-Cardonahttps://orcid.org/0000-0001-6083-5403Alejandro Martínez-Martínezhttps://orcid.org/0000-0003-3448-910XLuis Fernando Plenge-Tellecheahttps://orcid.org/0000-0002-1619-5004Edna Rico-Escobarhttps://orcid.org/0000-0002-0933-0220Esta revista está bajo una licencia de Creative Commons Reconocimiento 4.0 Internacional. Respirar 2023; 15 (3): 189ARTÍCULO ORIGINAL / E. Barrera-Ramírez, R.E. Garrido-Cardona, A. Martínez-Martínez, L.F. Plenge-Tellechea, E. Rico-EscobarDescelularización y regeneración de tráqueaISSN 2953-3414Materials and Methods: Porcine tracheas were decellularized by placing each one in the omentum of four Yorkshire pigs for regeneration in vivo. A trachea decellularized with triton (DT), decellularized with deoxycholate (DD), decellularized with deoxycho-late and reinforced with a polymer, and epithelial cells (DDR), and a cryopreserved na-tive (NC). After 8 days, the DD, NC and DDR were obtained; and on day 15, the DT. The evaluation was mechanically and histologically, performing the case analysis.Results: the decellularized tracheas preserved the integrity of the cartilage, with no me-chanical differences, except for the DDR with greater rigidity. The regenerated trache-as presented less rigidity, except the DDR, which also lost the epithelium and vascular-ity. The DT, DD showed non-respiratory epithelium, fibrosis and vasculogenesis with inflammation.Conclusions: the matrices retained their mechanical characteristics, in vivo regenera-tion offers advantages such as sterility, cell interaction, nutrients; it is simple, feasible and economical, but there is no control of cell growth and vascularization, and the tis-sues presented mechanical and histological alterations. The polymer prevented re-epi-thelialization and revascularization. This study opens the possibility of improving tissue engineering methodologies applied to tracheal tissue.

In situ isolation of nuclei or nuclear proteins from adherent cells: a simple, effective method with less cytoplasmic contamination

BACKGROUND: Isolation of nuclei or nuclear proteins is a prerequisite for western blot, nuclear proteome profiling, and other evaluations of nuclear proteins. Here, we developed a simple method for in situ isolation of nuclei or nuclear proteins by in situ removing the extranuclear part of adherent cells via a classical nonionic detergent triton X-100. RESULTS: First, the feasibility of our method was confirmed by confocal microscopy, atomic force microscopy, scanning electron microscopy, dynamic light scattering, immunofluorescence imaging, and time-lapse dynamic observation. Next, the optimal concentration range (approximately 0.1-1% for ~ 10 min) of triton X-100 and the optimal treatment time (< 30 min) of 0.1-1% Triton X-100 for our method were determined via western blotting of eight extra-/ intra-nuclear proteins. Subsequently, the effectiveness, sensitivity, and cytoplasmic contamination of our method were tested by investigating the levels of phosphorylated p65 (a NF-κB subunit) in the nuclei of endothelial or tumor cells treated with/without lipopolysaccharide (LPS) via western blotting and by comparing with a commercial nuclear protein extraction kit (a classical detergent-based method). The data show that compared with the commercial kit our method obtained a higher yield of total nuclear proteins, a higher pP65 level in both control and LPS groups, and much lower content of GAPDH (as a reference for cytoplasmic contamination) in nuclei. CONCLUSIONS: The in situ isolation of nuclei or nuclear proteins from adherent cells in this study is a simple, effective method with less cytoplasmic contamination. This method/strategy has the potential of improving the quality of downstream evaluations including western blotting and proteomic profiling.

Enhanced production of erythritol and mannitol by Yarrowia lipolytica in media containing surfactants

Abstract Various chemical compounds, including surfactants, when introduced to culture media may increase the permeability of cellular membranes and thereby affect the quantity of metabolites excreted by cells. The aim of the present study was to evaluate the impact of detergents including Triton X-100, Span 20 and Tween 80 on erythritol production from glycerol by Yarrowia lipolytica Wratislavia K1 in a shake-flask experiment, batch and fed-batch cultures. When Span 20 was added to a fed-batch culture with glycerol as a carbon source (300 g L-1), erythritol production increased by 15% compared to the culture without the surfactant where it reached 142 g L-1 after 5 days, which corresponded to 0.47 g g-1 yield and productivity of 1.1 g L-1 h-1. Therefore, it was concluded that Span 20 considerably enhanced the production of this polyol from glycerol.

Preparation of a decellularized scaffold derived from human liver tissue / 南方医科大学学报

<p><b>OBJECTIVE</b>To develop a method for preparing a decellularized scaffold based on human liver tissue.</p><p><b>METHODS</b>A surgical specimen of the left lateral lobe of the liver was obtained from a patients with hepatic hemangioma. The decellularization process was performed by repeated freezing-thawing, sequential perfusion with 0.01% SDS, 0.1% SDS and 1% Triton X-100 through the portal vein, and sterilization with peracetic acid. L-02 cells were then engrafted onto the decellularized liver scaffold.</p><p><b>RESULTS</b>HE staining, DAPI staining and scanning electron microscopy all verified the absence of residual cellular components in the decellularized scaffold. The residual DNA content in the decellularized scaffolds was 25.3∓14.6 ng/mg (dry weight), which was less than 1% of the total DNA content in a fresh human liver. Immunohistochemistry demonstrated that type I and IV collagens, fibronectin and elastin were all retained in the scaffold. The engrafted L-02 cells survived well on the scaffold with active proliferation and expressed albumin and G6pc.</p><p><b>CONCLUSION</b>It is feasible to prepare decellularized scaffolds using surgical specimens of human liver, which can be a new approach to constructing a tissue-engineered liver for clinical purposes.</p>

Optimization of Protein Extraction for Lichen Thalli

Lichen-forming fungal proteins have been seldom searched due to many difficulties in their extraction. Phenols, quinones, proteases, and other components released during cell disruption have been known to be the greatest challenges related to protein extraction from lichens. To overcome these problems and maintain good electrophoretic resolution and high protein concentration, an extraction buffer containing polyvinylpolypyrrolidone, ascorbic acid, Triton X-100, polyethylene glycol, proteinase, and oxidase inhibitors in sodium phosphate buffer was developed. This extraction buffer showed high efficiency for all lichen species tested in the study.

Obturation Over an S1 ProTaper Instrument Fragment in a Mandibular Molar with Three Years of Follow-up

This case report describes root canal filling performed over a large S1 ProTaper file fragment in a second mandibular molar with irreversible pulpitis. An S1 ProTaper file was fractured during the instrumentation of the mesiobuccal canal. Approximately 10 mm of file fragment remained in the apical and middle thirds of the canal. The obturation was performed over this fragment using thermomechanically compacted gutta-percha and sealer. Radiographic findings and the absence of clinical signs and symptoms at 3-year follow up indicated successful treatment. Cone-beam computed tomography images revealed absence of periapical lesion and details of intracanal file fragment related to root fillings and apex morphology. In this case, the presence of a large intracanal fractured instrument did not have a negative impact on the endodontic prognosis during the follow up evaluation period.

Este relato de caso descreve a obturação do canal radicular realizada sobre um grande fragmento da lima ProTaper S1 em um segundo molar inferior com pulpite irreversível. Uma lima ProTaper S1 fraturou durante a instrumentação do canal mésio-vestibular. Aproximadamente 10 mm de remanescente do fragmento da lima permaneceu nos terços apical e médio do canal. A obturação foi realizada sobre este fragmento usando guta-percha compactada termomecanicamente e cimento endodôntico. Achados radiográficos e ausência de sinais e sintomas clínicos após 3 anos de acompanhamento indicaram o sucesso do tratamento. Imagens de tomografia computadorizada de feixes cônicos revelaram a ausência de lesão periapical e detalhes do fragmento da lima intracanal relacionados à obturação do canal radicular e à morfologia do ápice. Neste caso, a presença de grande instrumento fraturado intracanal não teve impacto negativo no prognóstico endodôntico durante o período de acompanhamento.

Breast cancer in Latin America: global burden, patterns, and risk factors / Cáncer de mama en América Latina: carga, patrones y factores de riesgo

Breast cancer is a major public health problem in Latin America (LA) and the most common form of cancer among women. An important variability according to ethnicity/race with respect to incidence/mortality, clinical characteristics, and prognosis is observed throughout LA. In addition, women are more likely to develop breast cancer (BC) at younger age and to be diagnosed at an advanced stage compared to western women. While little is known about specific risk factors, changes in reproductive pattern (parity, breastfeeding) and lifestyle factors including sedentary behaviours, unhealthy diet, and alcohol intake may contribute to the increase of BC incidence. In this paper we give an overview of the burden and patterns of BC, review the leading causes of BC and discuss the possible ways to improve BC prevention and control in LA.

El cáncer de mama (CaMa) es uno de los mayores problemas de salud pública en América Latina (AL) y el cáncer más frecuente en mujeres. Se observa una importante variabilidad en la incidencia/mortalidad, las características clínicas y el pronóstico según la etnia/raza a lo largo de AL. Además, las mujeres latinoamericanas son más propensas a desarrollar CaMa en edades más tempranas y a ser diagnosticadas en una etapa más avanzada, comparando con mujeres occidentales. Aunque poco se sabe sobre sus factores de riesgo específicos, cambios en los patrones reproductivos (paridad y lactancia) y estilos de vida, incluyendo los hábitos sedentarios, las dietas poco saludables y el consumo de alcohol, podrían contribuir al incremento de la incidencia del CaMa. En este artículo se da una visión general de la carga y los patrones del CaMa, se revisan las causas principales del CaMa y se discuten posibles vías para mejorar la prevención y el control del CaMa en AL.

Acquirement and evaluation of murine ventricular extracellular matrix / 生理学报

Cardiac extracellular matrix (ECM), generated from the process of decellularization, has been widely considered as an ideal source of biological scaffolds. However, current ECM preparations are generally difficult to be applied to generate cardiac tissue. Our research was aimed to improve decellularization protocols to prepare cardiac ECM slices. Adult murine ventricular tissues were embedded in low melting agarose and cut into 300 μm slices, and then were divided randomly into three groups: normal cardiac tissue, SDS treated group (0.1% SDS) and SDS+Triton X-100 treated group (0.1% SDS+0.5% Triton X-100). Total RNA content and protein content quantification, HE staining and immunostaining were used to evaluate the removal of cell components and preservation of vital ECM components. Furthermore, murine embryonic stem cell-derived cardiomyocytes (mES-CMs) and mouse embryonic fibroblasts (MEFs) were co-cultured with ECM slices to evaluate biocompatibility. The relative residual RNA and protein contents of ECM slices significantly decreased after decellularization. HE staining showed that SDS+Triton X-100 treatment better destroyed cellular structure and removed nuclei of ECM slices, compared with SDS treatment. Immunostaining showed that collagen IV and laminin were better preserved and presented better similarity to original cardiac tissue in ECM slices acquired by SDS+Triton X-100 treatment. However, collagen IV and laminin were significantly decreased and arranged disorderly in SDS treated group. We observed effective survival (≥ 12 days) of MEFs and mES-CMs on ECM slices acquired by SDS+Triton X-100 treatment, and signs of integration, whereas those signs were not found in SDS treated group. We concluded that, compared with traditional SDS method, new combined protocol (SDS+Triton X-100) generated ECM slices with better component and structural preservation, as well as better biocompatibility.

Experimental study on removal melanin granules from acellular dermal matrix of giant nevus / 中华整形外科杂志

<p><b>OBJECTIVE</b>To study the possibility of removal melanin granules from autogenic acellular dermal matrix of giant nevus tissue by H2O2 bleaching technique.</p><p><b>METHODS</b>A total of 32 skin specimens (0.5 cm x 0.5 cm) from giant nevus tissue and 1 piece (0.5 cm x 0.5 cm) of normal skin were obtained from the surgical removal. One giant nevus tissue was chosen as control. The others and the normal skin tissue were treated with solution of 0.25% Dispase II for digestion for 24 hours under normal temperature to remove epidermis. Then each piece was immerged into solution of 0.5% Triton X-100 for digestion for 48 hours in normal temperature. One giant nevus tissue and the normal skin tissue were chosen as control. The others were immerged into solution of different concentrations of H2O2, treated under different temperature and lasting for different period. Lastly, all specimens were treated with HE staining, immunohistochemical staining, light microscopy and so on.</p><p><b>RESULTS</b>After giant nevus tissues were treated with solution of 0.25% Dispase II and immerged into solution of 0.5% Triton X-100 in normal temperature, nevus cells and all other cellular components of pigmented nevus tissues can be effectively removed, there were the cavities left by removal of cells without any residual cell debris, but still remaining part of pigment. Then each specimen were immerged into solution of different concentrations of H2O2, under different temperature and lasting for different period which can remove residual melanin granules. In solution of 3% H2O2 for 36 h under 37 degrees C, can remove all the melanin particles, the content of collagen type I in the obtained specimen was not changed. Collagen fibers were uniform in thickness, regular in arrangement with no obvious degeneration.</p><p><b>CONCLUSIONS</b>With solution of 0.25% Dispase II and solution of 0.5% Triton X-100 in normal temperature, all cells in nevus tissue can be removed effectively. Further treatment with 3% H2O2 at 37 degrees C for 36 h can remove all the melanin particles, while collagen type I has no obvious change. The preparation of acellular dermal matrix of the giant nevus may possibly be applied as autologous tissue implant to repair tissue defects.</p>

Effect of Trypsin on Physico-dynamic and Histological Changes after Decellularization of Bovine Pericardium / 대한흉부외과학회지

BACKGROUND: Various decellularization methods have been studied in order to develop tissue graft which is less immunogenic and more durable. This study was performed to investigate the physico-dynamic and histological effect of trypsin pretreatment on decellularization protocols. MATERIAL AND METHOD: Two groups of bovine pericardium specimen each underwent decellularization process based on SDS and Triton X-100 or N-lauroylsarcosinate and Triton X-100. Two more groups additionally underwent pretreatment with 0.1% Trypsin/0.1% EDTA. After decellularization process, mechanical tensile strength was tested, then iomechanical test of permeability and compliance was tested before and after fatigue test. Light microscopy and electron microscopy was performed to observe histological findings. RESULT: There was no difference in mechanical tensile strength between groups, but permeability and compliance was decreased in trypsin pretreated groups. Light microscopic and electron microscopic findings revealed damage of the extracellular matrix in trypsin pretreated groups and in groups which underwent the fatigue test also. CONCLUSION: Trypsin pretreatment in decellularizing process of bovine pericardium damages extracellular matrix and increases permeability and compliance of the bovine pericardium, but did not decrease tensile strength. Further studies are needed to use enzymatic treatments in decellularization protocols.

Bacteriocin from Purple Nonsulfur Phototrophic Bacteria, Rhodobacter capsulatus

To find whether productivity of bacteriocin is controlled between different species under unusual cultural conditions, we used Rhodobacter capsulatus ATCC 17016 as a producer and Rhodopseudomonas palustris ATCC 17003 as an indicator. Rhodobacter capsulatus was cultured under aerobic conditions in the dark in Lascelles medium containing 0.3% Triton X-100. As a result, bacteriocin productivity increased enormously. The optimal pH range of bacteriocin production was 6~7.8. Through partial purification of bacteriocin, the molecular weight was roughly estimated at 14 kDa. Plasmid had no influence on bacteriocin production by Rhodobacter capsulatus. Our findings indicate that culture conditions affect bacteriocin productivity between more distantly related species, and bacteriocin of Rhodobacter capsulatus is not encoded by a plasmid.

Construction of an acellular porcine aortic valve / 南方医科大学学报

<p><b>OBJECTIVE</b>To prepare a porcine aortic valve (PAV) free of the cellular components.</p><p><b>METHODS</b>The cellular components of porcine PAV were completely removed using trypsin and Triton X-100, and the acellular PAV was examined microscopically with HE staining with its physical and chemical properties assessed. Transmission electron microscopy was used to observe the integrity of the collagen and elastin and the DNA contents in the PAV was detected to confirm the total removal of the cellular components. With the fresh PAV as the control, small pieces of the acellular PAV were implanted into the subcutaneous tissues of 4 rabbits, and 4 weeks after the implantation, the implants were harvested for microscopic observation.</p><p><b>RESULTS</b>The cellular components were effectively removed from the cusps and roots of the PAV by trypsin and TritonX-100, with marked soluble protein loss [(0.24-/+0.04)% vs (0.48-/+0.12)%] and significantly increased water content [(92.2-/+1.5)% vs (89.2-/+1.6)%]. The acellular PAV still maintained good fibrous scaffold structure and the shrinkage temperature and tension at fracture underwent no significantly changes [(67.9-/+1.0) degrees celsius; vs (68.8-/+0.8) degrees celsius; and (489.3-/+19.0) g/mm2 vs (540.7-/+19.5) g/mm2, respectively]. The PAVs implanted in rabbits showed only mild tissue reaction with a few infiltrating neutrophils, lymphocytes and plasmocytes observed 4 weeks later. The accelular PAV caused obviously milder inflammatory reactions than fresh PAV.</p><p><b>CONCLUSIONS</b>The acellular PAV prepared by treatment with trypsin and Triton X-100 retains good fibrous scaffold structure and mechanical strength with low antigenicity.</p>

Determination of twenty pesticide residues in Viscum coloratum by gas chromatography using cloud-point extraction / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a gas chromatography method for simultaneous determination of organochlorine and pyrethroid pesticide residues in Viscum coloratum by cloud-point extraction (CPE).</p><p><b>METHOD</b>Pesticides were extracted with the non-ionic surfactant Triton X-100. The apparatus was gas chromatography with electron capture detector and the separation was performed on an Hp-5 column. The pesticide residues were calculated by external standard method.</p><p><b>RESULT</b>Good linear relation was obtained over the range of 5-500 microg L(-1) for organochlorine and 10-1,000 microg L(-1) for pyrethroid. The limits of detection was 1.5-7.5 microg kg(-1). The average recoveries of organochlorine and pyrethroid were 74.15% -111.6% with corresponding RSD of 4.0% -9.1%.</p><p><b>CONCLUSION</b>The sample and rapid method was applied to pesticide residues determination.</p>

Enhanced crude oil biodegradation and rhamnolipid production by Pseudomonas stutzeri strain G11 in the presence of Tween-80 and Triton X-100.

In this study, the growth of sixty-one bacterial strains in crude oil were determined spectrophotometrically at 620 nm. Pseudomonas aeruginosa G1, Pseudomonas fluorescens G6, Pseudomonas stutzeri G11 and Pseudomonas putida G15 were chosen for the study based on the efficiency of crude oil utilisation. At 1% (v/v) crude oil concentration, P. stutzeri G11 strain degraded a maximum of 69%. The percentage of degradation by the P. stutzeri G11 strain decreased from 69% to 59% as the concentration of crude oil was increased from 1% (v/v) to 2.5% (v/v). Strain G11 was selected to determine the effects of surfactants (Tween-80 and TritonX-100) on the biodegradation of crude oil. While strain G11 showed 76% degradation at mineral salts medium (MSM) containing 1% (v/v) crude oil + 1% (v/v) TritonX-100, it showed 61% degradation at MSM containing 2.5% (v/v) crude oil + 2.5% (v/v) TritonX-100. Also, degradation rate of this strain was 96% in the presence of 1% (v/v) crude oil + 1% (v/v) Tween-80, while degradation rate was 48% in the presence of 25% (v/v) crude oil+ 2.5% (v/v) Tween-80. Additionally, we investigated the rhamnolipid production of P. stutzeri G11 strain both in crude oil and in crude oil + two different surfactants (TritonX-100 and Tween-80, separately). These results suggest that surfactants have improved both crude oil degradation and rhamnolipid production and the degradation rates have depended very much on the chemical structure of surfactants.

Study on an Effective Decellularization Technique for Cardiac Valve, Arterial Wall and Pericardium Xenographs: Optimization of Decellularization / 대한흉부외과학회지

BACKGROUND: We attempted to reproduce a previously reported method that is known to be effective for decellularization, and we sought to find the optimal condition for decellularization by introducing some modificationsto this method. MATERIAL AND METHOD: Porcine semilunar valves, arterial walls and pericardium were processed for decellularization with using a variety of combinations and concentrations of decellularizing agents under different conditions of temperature, osmolarity and incubation time. The degree of decellularization and the preservation of the extracellular matrix wereevaluated by staining with hematoxylin and eosin and with alpha-Gal and DAPI in some of the decellularized tissues. RESULT: Decellularization was achieved in the specimens that were treated with sodium deoxycholate, sodium dodesyl sulfate, Triton X-100 and sodium dodesyl sulfate with Triton X-100 as single-step methods, and this was also achieved in the specimens that were treated with hypotonic solution --> Triton X-100 --> sodium dodesyl sulfate, sodium deoxycholate --> hypotonic solution --> sodium dodesyl sulfate, and hypotonic solution sodium dodesyl sulfate as multi-step methods. CONCLUSION: Considering the number and the amount of the chemicals that were used, the incubation time and the degree of damage to the extracellular matrix, a single-step method with sodium dodesyl sulfate and Triton X-100 and a multi-step method with hypotonic solution followed by sodium dodesyl sulfate were both relatively optimal methods for decellularization in this study.

Depressed CCL5 Expression in Human Pulmonary Tuberculosis

CCL5/regulated on activation, normal T expressed and secreted production (RANTES) is a principal CC chemokine, and can activate macrophages and Th1 lymphocytes, however, little is known about the CCL5 profiles associated with active tuberculosis (TB). In this study, we investigated the production of CCL5 by the peripheral blood mononuclear cells (PBMCs) of patients with active pulmonary TB after stimulation with Triton X-100 soluble proteins (TSP) or the 30-kDa antigen. The profiles of cytokines/chemokines [CXCL8/interleukin (IL)-8, IL-12 p40, and interferon (IFN)-gamma] were also examined by PBMCs from TB patients, and compared with those obtained from healthy tuberculin reactors (HTR). Concordant with earlier studies, IFN-gamma production was significantly depressed in the PBMCs from TB patients compared with those from HTR. In addition, the CCL5, but not CXCL8, levels in the PBMCs from TB patients were significantly depressed after stimulation for 18 hr compared to those in the PBMCs from HTRs. The CCL5 release was not significantly correlated with the release of IFN-gamma in the cells from TB patients and HTRs. Further, inhibitor studies show that the 30-kDa- or TSP-induced CCL5 mRNA expression is sensitive to inhibitors of mitogen-activated protein kinase kinase (MEK) 1/2 and Janus kinase (JAK) 2, but not p38, pathway activation, suggesting a MEK1/2- or JAK2-based mechanism is responsible for modulating of the CCL5 expression in human PBMCs. Collectively, these data suggest that TB patients show depressed production of CCL5 secretion, which can be modulated by MEK- and JAK2-based transcriptional regulatory mechanisms, in response to the mycobacterial antigens.

Effect of polyoxyl ether analogous surfactants on the activity of cytochromes P450 3A in rats in vivo / 药学学报

To evaluate the effects of p-octyl polyethylene glycol phenyl ether (Triton X-100), polyoxyl 35 caster oil (EL35) and polyoxyl 40 hydrogenated caster oil (RH40) on the activity of Cytochrome P450 3A (CYP3 As) in vivo. Rats were administered with saline, ketoconazole (75 mg x kg(-1) x d(-1)), Triton X-100 (30 mg x kg(-1) x d(-1)), EL35 (150 mg x kg(-1) x d(-1)) and RH40 (150 mg x kg(-1) x d(-1)) intragastrically for 5 consecutive days, and then given midazolam 10 mg x kg(-1) 20 min after the last treatment of ketoconazole or three surfactants with the same dose through duodenal administration. Pharmacokinetics parameters for midazolam and its metabolite 1'-hydroxymidazolam were estimated from the plasma concentration-time data by a noncompartmental approach. The results showed that multiple dose administration of Triton X-100, EL35 and RH40 decreased the ratios of 1'-hydroxymidazolam and midazolam AUC0-infinity from 1.14 to 0.90, 1.03 and 0.64, respectively. In contrast, multiple dose administration of ketoconazole caused the ratios of 1'-hydroxymidazolam and midazolam a significant decrease to 0.50. This study indicated that Triton X-100 and EL35 would have no inhibition on CYP3A, while RH40 had significant inhibition on CYP3A. Therefore, RH40 might be used to prepare drug formulations in pharmaceutical industry and would increase the bioavailability of some drugs transformed by CYP3As and further lead to significant clinical pharmacologic effects.

Elevated Levels of Interferon-inducible Protein-10 (IP)-10/CXCL10, but not of Interferon-gamma, in Patients with Pulmonary Tuberculosis

Mycobacterial strains are potent inducers of cytokines/chemokines by mononuclear phagocytes, which constitute an important cellular component of the first line of defense in the innate immune system. Interferon (IFN)-gamma-inducible protein (IP-10 or CXCL10) is a potent chemoattractant; however, little is known about the IP-10 profiles attributable to the Th1 regulation associated with active tuberculosis (TB). In this study, we investigated the production of IP-10, interleukin (IL)-12 p40, and IFN-gamma by the peripheral blood mononuclear cells (PBMCs) of patients with active pulmonary TB in response to in vitro stimulation with Triton X-100 soluble proteins (TSPs) or the 30-kDa antigen. The TSP antigens used in the present study were isolated and purified from Mycobacterium tuberculosis H37Rv (virulent strain), M. tuberculosis H37Ra (avirulent strain), and Mycobacterium bovis BCG. The results were compared with those obtained for healthy tuberculin reactors (HTRs). Concordant with earlier studies, IFN-gamma production was significantly depressed in the PBMCs from TB patients compared with those in the HTR group. However, the IP-10 levels in the PBMCs from TB patients were significantly elevated 18 h after stimulation compared to those in the PBMCs from HTRs. IP-10 release was correlated in a significant manner with the release of IFN-gamma in the HTRs, but this was not the case for the TB patients. Collectively, these data suggest that TB patients show altered regulation of Th1-driving cytokine and chemokine production in response to a variety of mycobacterial antigens.

Hypolipidemic and antihyperlipidemic effects of Lagenaria siceraria (Mol.) fruit extracts.

Bottle gourd [(Lagenaria siceraria (Mol.) Stand.] fruit is ascribed with many therapeutic effects. The present study was undertaken to explore the antihyperlipidemic effect of four different extracts viz. petroleum ether, chloroform, alcoholic and aqueous extracts from bottle gourd in Triton-induced hyperlipidemic rats and their hypolipidemic effects in normocholesteremic rats. The study is comprised preliminary phytochemical screening of the extracts. Oral administration of the extracts, at doses of 200 and 400 mg/kg body weight in rats, dose-dependently inhibited the total cholesterol, triglycerides, low-density lipoproteins level, and significantly increased the high density lipoproteins level. However, petroleum ether extract did not show the significant effects. Both the chloroform and alcoholic extract exhibited more significant effects in lowering total cholesterol, triglycerides and low density lipoproteins along with increase in HDL as compared to the others. Preliminary phytochemical screening revealed the presence of flavonoids, sterols, cucurbitacin saponins, polyphenolics, proteins, and carbohydrates. The results obtained suggest marked antihyperlipidemic and hypolipidemic activity of the extracts.

Kinetics and mechanisms of cholesterol esterase inhibition by cardiovascular drugs in vitro.

Cardiovascular drugs such as lovastatin, simvastatin, amlodipine besylate, nifedipine, and hydralazine hydrochloride inhibit cholesterol esterase (CEase) in vitro. In the present paper, an attempt was made to determine kinetically the reaction mechanism for CEase inhibition by these drugs. The inhibition constant, Ki, for the mixed-type inhibition of CEase by these drugs in the presence of triton-X-100 or taurochloate were measured. Moreover, the pKi values were correlated with the molecular weights of these drugs. In conclusion, the fact that these drugs lower cholesterol levels in the plasma low-density lipoprotein may be partially due to the CEase inhibition by these drugs.