Maintenance and characterization of multipotent mesenchymal stem cells isolated from canine umbilical cord matrix by collagenase digestion.

Multipotent mesenchymal stem cells have been considered as a novel clinical approach for cell therapy and regenerative medicine. In this study, mesenchymal stem cells (MSCs) were successfully isolated from canine umbilical cord matrix (cUCM; also referred to as Wharton's Jelly) by collagenase digestion and further characterized for multipotent properties associated with MSCs. Our cUCM-derived MSCs (cUCM-MSCs) were plastic adherent, spindle-shaped and fibroblast-like cells, maintaining expression of pluripotency markers such as Oct3/4, Nanog, Sox-2 and SSEA-4 as well as normal chromosomal number during a long-term proliferative culture. The cells expressed MSCs-specific surface markers, including CD44, CD90, CD105, and CD184, but did not CD29, CD33, CD34, and CD45. More importantly, cUCM-MSCs could differentiate into mesodermal (adipocyte, osteocyte and chondrocyte) and ectodermal (neuronal cell) cell lineages. These results imply that collagenase digestion would be a highly effective way to isolate multipotent MSCs in abundant amounts.

[Heterologous allogeneic blood transfusion as a risk factor associated with the adverse course of the surgical wound in primary hip surgery]. / Hemotransfusión alogénica heteróloga como factor de riesgo asociado con la evolución adversa de la herida quirúrgica en cirugía primaria de cadera.

BACKGROUND: Due to its immunomodulating effects mediated by leukocytes and interleukins, heterologous allogeneic blood transfusion has been considered as a risk factor for both morbidity and mortality in patients undergoing orthopedic surgery, including hip surgery. This research analyzed whether heterologous allogeneic blood transfusion is a risk factor associated with the adverse course or complication of the surgical wound in patients undergoing primary hip surgery due to fracture at a general hospital in 2008-2009. MATERIAL AND METHODS: Forty-nine patients who had a complication (cases) and 207 with no complications (controls) were identified and both groups were compared with a bivariate and multivariate analysis, and demographic and clinical data, including having undergone blood transfusion or not. RESULTS: Not having received a blood transfusion was identified as a variable that reduced the risk of surgical wound complications (OR = 0.05, 95% confidence interval [CI 95%] 0.0067 to 0.16; chi2 with p < 0.001). The multivariate model excluded as clinically significant variables the duration of surgery (OR = 1.01, CI 95% 0.99 to 1.02; p = 0.12) and certain chronic conditions (OR = 0.54, CI 95% 0.13 to 2.24 for diabetes mellitus, OR = 1.16, CI 95% 0.29 to 4.60 for chronic hypertension, OR = 1.21, CI 95% 0.19 to 7.51 for various heart diseases). CONCLUSIONS: Not having received a blood transfusion reduced 95% the risk of surgi- cal wound complications. Neither the duration of surgery nor a specific comorbid condition were associated with the former event.

Effect of ginsenosides, active components of ginseng, on capsaicin-induced pain-related behavior.

Our recent study demonstrated that ginsenosides had antinociceptive effects by reducing some types of pain-related behavior in mice (Yoon et al., 1998. Ginsenosides induce differential antinociception and inhibit substance P-induced nociceptive response in mice. Life Science 62, PL319-PL325). In the present study we further investigated whether ginsenosides produce antinociceptive effects through an action at central or peripheral site(s) and whether these effects are mediated by the opioid system. Intraperitoneally injected ginsenosides suppressed in a dose-dependent manner the pain-related behavior produced by capsaicin injection into the plantar surface of the hind paw; the ED(50) was 49 mg/kg [26-92 mg/kg, 95% confidence interval (C.I.)]. Intrathecally or intracerebroventricularly administered ginsenosides also suppressed the capsaicin-induced pain-related behavior in a dose-dependent manner; the ED(50)s were 1.72 mg/kg (0.8-3.72 mg/kg, 95% C.I.) and 1. 48 mg/kg (0.8-2.6 mg/kg, 95% C.I.), respectively. On the other hand, subcutaneously injected ginsenosides to the plantar surface prior to the capsaicin injection did not alter the pain-related behavior. Naloxone pretreatment was without effect in blocking the antinociceptive effect of intrathecally administered ginsenosides. Intraperitoneally injected ginsenosides also did not significantly affect the motor response of animals. These results suggest that ginsenosides produce antinociceptive effects through their action at the spinal and/or supraspinal site(s), not at nociceptors in the periphery. In addition, the results suggest that the antinociceptive effects are not mediated by opioid receptors.

Ginsenosides inhibit capsaicin-activated channel in rat sensory neurons.

Ginsenosides isolated from ginseng are biologically active components. In this study, whole-cell and inside-out configurations of patch clamp technique had been used to test the effect of ginsenosides on the capsaicin-activated channels in cultured small diameter sensory neurons of young rat. Ginsenosides (100 microg/ml) decreased the amplitude of capsaicin-activated currents by 78.2% in whole cell mode. Similarly, ginsenosides decreased capsaicin-activated single-channel activities in a dose-dependent manner in inside-out patches. These results indicate that ginsenosides might directly block capsaicin-activated channels, resulting in attenuation of the currents in rat sensory neurons.

Preparation of monoclonal antibody against ginsenoside Rf and its enzyme immunoassay.

A rapid and sensitive indirect competitive enzyme immunoassay method has been developed for quantitating ginsenoside Rf (Rf) in crude total Panax ginseng saponins and in rat plasma using high titer mouse monoclonal antibody (mAb) raised against a conjugate of Rf and bovine serum albumin (BSA). The isotype of mAb against Rf was IgG3 with a K chain. The presence of Rf inhibited the binding of the mouse anti-Rf mAb to a Rf-BSA solid phase coating antigen. The working range was 0.01-10 ng/assay and detection limits were 20 pg in various ginseng extract fractions or 34 pg in rat plasma per assay. The anti-Rf mAb cross-reacted with ginsenoside Rg2 by 57.5%, but not with other ginsenosides. However, this anti-Rf mAb did not cross-react with BSA or cellubiose, which is a carbohydrate component of Rf. Using this standard curve, we could measure the amount of Rf in ginseng total extract, ginseng total saponins, protopanaxadiol saponins, and propanaxatriol saponins. We could also measure the amount of Rf in rat plasma after the oral administration of Rf and found that Rf reached a maximum level in rat plasma after 16 h. These results indicate that the anti-Rf mAb could be useful for the quantitation of Rf in crude ginseng fractions and in body fluids.

Characterization and expression of a novel alternatively spliced human angiopoietin-2.

Angiopoietin-2 (Ang2) is a naturally occurring antagonist of angiopoietin-1 (Ang1) that competes for binding to the Tie2 receptor and blocks Ang1-induced Tie2 autophosphorylation during vasculogenesis. Using the polymerase chain reaction, we isolated a cDNA encoding a novel shorter form of Ang2 from human umbilical vein endothelial cell cDNA and have designated it angiopoietin-2(443) (Ang2(443)), because it contains 443 amino acids. Part of the coiled-coil domain (amino acids 96-148) is absent in Ang2(443) because of alternative splicing of the gene. Like Ang2, recombinant Ang2(443) expressed in COS-7 cells is secreted as a glycosylated homodimeric protein. Recombinant Ang2(443) binds to the Tie2 receptor but does not induce Tie2 phosphorylation. Pre-occupation of Ang2(443) on Tie2 inhibits Ang1 or Ang2 binding and inhibits Ang1-induced phosphorylation. Expression of Ang2(443) mRNA is detectable in primary endothelial cells, several nonendothelial tumor cell lines, and primary tumor tissues. Interestingly, two cervical carcinoma cell lines express relatively moderate levels of Ang2(443) mRNA and protein. Macrophages express mainly Ang2 mRNA, but the expression of Ang2(443) mRNA is temporarily up-regulated during macrophage differentiation. These results suggest that Ang2(443) is a functional antagonist of Ang1 and could be an important regulator of angiogenesis during some tumorigenic and inflammatory processes.

[Acute pesticide poisoning]. / Intoxicación aguda por plaguicidas.

OBJECTIVE: To describe the epidemiologic pattern of acute pesticide poisoning (APP) in a general hospital in Merida, Yucatan, Mexico. MATERIAL AND METHODS: From 1994 to 1998, 33 patients 13 years of age or older with diagnosis of APP were studied. Descriptive statistics were used to analyze information. RESULTS: Males were frequently affected (82%), specially those coming from rural areas (60%). The mean age of the group was 34 +/- 15.8 years. In 79% of the cases, pesticides were used to commit suicide and 33% of poisoning cases were due to organophospate pesticides. The mortality rate was 12%. CONCLUSIONS: In this small sample, acute poisoning from pesticides in the agricultural setting may be underestimated, since it was less frequent than in the general population. APP was more commonly used by indigent people to commit suicide.

Ginsenosides that produce differential antinociception in mice.

Ginsenoside Rc, Rd, and Re induced antinociception in writhing and formalin tests among five representative ginsenosides: Rb1, Rc, Rd, Re, and Rg1. However, these ginsenosides had no effect in the tail-flick test. The antinociceptive effects induced by three ginsenosides were dose dependent. ED50 was 20.5 (7.3-57.4 mg/kg) for Rc, 17 (11.0-27.6 mg/kg) for Rd, and 3.5 (1-12 mg/ kg) for Re in the writhing test and 62 (42-90 mg/kg) for Rc, 45 (20.5-99.0 mg/kg) for Rd, and 82 (48-139 mg/kg) for Re in the second phase of the formalin test. The antinociceptive effects were not blocked by the opioid receptor antagonist naloxone in the writhing and formalin tests. These three ginsenosides did not affect motor function. Ginsenoside Rc and Rd induced hypothermia for 30 to 60 min, and ginsenoside Rc induced hyperthemia after 150 min of treatment at doses of 100 mg/kg. These results suggest that ginsenosides such as Rc, Rd, or Re inhibit mainly chemogenic pain rather than thermal pain by the nonopioid system in mice.

[Epidemiology of preeclampsia-eclampsia at the General Hospital O'Horán. (Epidemiology of preeclampsia-eclampsia of a sample, in Yucatan)]. / Epidemiología de la preeclampsia-eclampsia en el Hospital General O'Horán. (Epidemiología de la preeclampsia-eclampsia de una muestra, en Yucatán).

The objective was to describe the epidemiology of preeclampsia-eclampsia (P-E) at the Hospital General O'Horán (HGOH) in Mérida, Yucatán, México, from 1995 to 1998. Patients with a discharge diagnosis of P-E were included. Their demographic and clinical data were ascertained. To analyze information, descriptive statistics were used. There were 143 patients. Preeclampsia was documented in 41% and eclampsia in 59%. The mean age of the group was 24.4 +/- 7.3 years. It was found that 76% came from rural area. In 79% schooling was no more than elementary education. Seventy five per cent were married. Sexual life began at a mean age of 18.8 +/- 4.3 years. There was no prenatal care in 27% of the cases. Fifty five per cent were primigravida and 43% multigravida. Nuliparity was documented in 52%. Two o more parities were documented in 48%. Complications were seen in 30%. Overall mortality rate was 5%, more frequent eclamptic patients. At the HGOH, P-E was frequently documented in women with both low socioeconomic status and fewer years of schooling. Prenatal care was also irregular. Clinical evolution was satisfactory in most of them, and the mortality rate was low, although it usually occurred in young eclamptic women.

Determination of ginsenoside Rf and Rg2 from Panax ginseng using enzyme immunoassay.

We have developed an enzyme immunoassay (EIA) to quantify trace amounts of ginsenoside Rf (Rf), one of the glycosides of protopanaxatriol from Panax ginseng. A carrier protein of bovine serum albumin (BSA) was coupled to the carbohydrate component of Rf using the periodate oxidation method. Antibodies were raised in rabbits using Rf-BSA conjugate as the immunogen and competitive indirect EIA was used for the determination of Rf. The working range was 0.01-10 ng per assay. The anti-Rf antiserum cross-reacted with ginsenoside Rg2 (105%), which is also a component of Panax ginseng and has a very similar chemical structure to Rf. These results suggest that the anti-Rf antiserum could also be used for the quantitation of ginsenoside Rg2 as well as ginsenoside Rf. In a comparison of EIA and HPLC the linear regression equation and correlation coefficient for the two methods were y(EIA) = 1.31x (HPLC)-11.48 and 0.98, respectively.

Ginsenosides induce differential antinociception and inhibit substance P induced-nociceptive response in mice.

Ginsenosides are main pharmacoactive molecules of ginseng. The antinociceptive activity of ginsenosides after intrathecal (i.t.) injection was examined in formalin test. We also investigated the effects of ginsenosides on substance P (SP) induced-pain behaviors by i.t. treatment using mice. Pretreatment of ginsenosides by i.t. induced the inhibition of biting and licking of hind paw injected with 1% formalin with dose-dependent manner. The ED50 was 23 (19-28, 95% C.I.) microg/mouse for acute phase and 15 (9-23, 95% C.I.) microg/mouse for tonic phase. Interestingly, cotreatment of ginsenosides with SP also inhibited SP-induced pain behaviors (scratching, licking or biting of hind portion of body) with dose-dependent manner. The ED50 for the inhibition of SP-induced pain behavior by ginsenosides was 30 (11-85, 95% C.I.) microg/mouse. These results suggest that ginsenosides have antinociceptive activity in formalin test and this effect is due to blocking of SP-induced nociceptive information to postsynaptic site(s) at the spinal level.