Inhibitory effects of herbal decoction Ojeoksan on proliferation and migration in vascular smooth muscle cells.

The proliferation of vascular smooth muscle cells plays a crucial role in pathogenesis of cardiovascular disease. The principal objective of this study was to determine the effects of Ojeoksan (OJS) on human aortic smooth muscle cell (HASMC) proliferation induced by tumor necrosis factor α (TNF-aα). Thymidine incorporation after TNF-α treatment was increased and this effect was inhibited significantly by OJS treatment. HASMC proliferation and migration by kinetic live cell imaging were also reduced by treatment with OJS. TNF-α induced the expression of cyclins/cyclin-dependent kinases (CDKs) and reduced the expression of p21waf1/cip1/p27kip1. However, OJS also attenuated the expression of TNF-α-induced cell-cycle regulatory proteins. The results of Western blot analysis demonstrated that the TNF-α treated HASMC secreted gelatinases, probably including MMP-2/-9, which may be involved in the invasion and migration of HASMC. Additionally, OJS suppressed the mRNA expression levels of matrix metalloproteinase-2/-9 (MMP-2/-9) in a dose-dependent manner. OJS inhibited the production of TNF-α-induced hydrogen peroxide (H2O2) and the formation of DCF-sensitive intracellular reactive oxygen species (ROS). Further, OJS suppressed the nuclear translocation and phosphorylation of inhibitor of kappa B-α (IκB-α) of nuclear factor κB (NF-κB) under TNF-α conditions. Our results demonstrate that OJS exerts inhibitory effects on TNF-α-induced HASMC proliferation and migration, suggesting the involvement of the inhibition of both MMP-2 and MMP-9 expressions, and the downregulation of ROS/NF-κB signaling. Thus, herbal decoction OJS may be a possible therapeutic approach to the inhibition of cardiovascular disease including atherosclerosis.

Mantidis ootheca induces vascular relaxation through PI3K/AKT-mediated nitric oxide-cyclic GMP-protein kinase G signaling in endothelial cells.

Mantidis ootheca (Sang Piao Xiao) is well known mantis eggs in a foamy pouch. The purpose of the present study was to investigate the underlying cellular mechanisms of the nitric oxide (NO)-releasing property of the aqueous extract of Mantidis ootheca (AMO) in rat aorta and vascular endothelial cells. AMO was examined for its vascular relaxant effect in isolated phenylephrine-precontracted rat thoracic aortic rings. The roles of the nitric oxide (NO) signaling in the AMO-induced effects were tested in human umbilical vein endothelial cells (HUVECs). HUVEC treated with AMO produced higher amount of NO compared to control. However, AMO-induced increases in NO production were blocked by pretreatment with NG-nitro-L-arginine methylester (L-NAME) or wortmannin. AMO increased in phosphorylation levels of endothelial nitric oxide synthase (eNOS) and Akt in HUVECs, which were attenuated by a NOS and Akt inhibitors. In aortic ring, AMO-induced dose-dependent relaxation of phenylephrine-precontracted aorta was abolished by removal of functional endothelium. Pretreatment with L-NAME, 1H-[1,2,4]-oxadiazolo-[4,3-alpha]-quinoxalin-1-one (ODQ), and KT5823 inhibited the AMO-induced vasorelaxation. Similarly, wortmannin and LY-294002, an inhibitors of the phosphatidylinositol 3-kinase (PI3K), an upstream signaling molecule of eNOS, attenuated the AMO-induced vasorelaxation. Moreover, AMO-induced increases in cGMP production were blocked by pretreatment with L-NAME or ODQ. The vasorelaxant effect of AMO was attenuated by tetraethylammonium, 4-aminopyridine, and glibenclamide. We conclude that AMO relaxed vascular smooth muscle via endothelium-dependent activation of PI3K/Akt-mediated NO-cGMP-PKG signaling pathway and possible involvement of K+ channel.

Direct hospital costs of total laparoscopic hysterectomy compared with fast-track open hysterectomy at a tertiary hospital: a retrospective case-controlled study.

AIMS: To assess the direct intraoperative and postoperative costs in women undergoing total laparoscopic hysterectomy and fast-track open hysterectomy. METHODS: A retrospective review of the direct hospital-related costs in a matched cohort of women undergoing total laparoscopic hysterectomy (TLH) and fast-track open hysterectomy (FTOH) at a tertiary hospital. All costs were calculated, including the cost of advanced high-energy laparoscopic devices. The effect of the learning curve on cost in laparoscopic hysterectomy was also assessed, as was the hospital case-weighted cost, which was compared with the actual cost. RESULTS: Fifty women were included in each arm of the study. TLH had a higher intraoperative cost, but a lower postoperative cost than FTOH (AUD$3877 vs AUD$2776 P < 0.001, AUD$3965 vs AUD$6233 P < 0.001). The total cost of TLH was not different from FTOH (AUD$7842 vs AUD$9009 P = 0.068) and after a learning curve; TLH cost less than FTOH (AUD$6797 vs AUD$8647, P < 0.001). The use of high-energy devices did not impact on the cost benefit of TLH, and hospital case-weight-based funding correlated poorly with actual cost. CONCLUSION: Despite the use of fast-track recovery protocols, the cost of TLH is no different to FTOH and after a learning curve is cheaper than open hysterectomy. Judicious use of advanced energy devices does not impact on the cost, and hospital case-weight-based funding model in our hospital is inaccurate when compared to directly calculated hospital costs.

The viability and function of cryopreserved hepatocyte spheroids with different cryopreservation solutions.

We sought to evaluate the viability and function of cryopreserved hepatocyte spheroids using different cryopreservation solutions in order to elucidate the efficiency of cryopreservation. Hepatocytes isolated from a Sprague-Dawley rat were formed into spheroids by 24 hours of rotational culture. The spheroids were cryopreserved using a programmed linear freezer in a liquid nitrogen tank for 24 hours in four different cryopreservation solutions: UW solution (UW), William E media (WE), fetal bovine serum (FBS), and a mixture (MIX). After thawing, they were cultured for 4 days. With each hepatocyte spheroid, the viability using the MTT assay and hepatocyte-specific functions, such as ammonia clearance, urea nitrogen synthesis, and albumin secretion, were analyzed. The viabilities of cryopreserved hepatocyte spheroids were 64.8% +/- 10.2% (UW), 33.2% +/- 9.7% (WE), 69.3% +/- 8.7% (FBS), and 48.4% +/- 15.5% (MIX). Ammonia clearance of spheroids cyropreserved in UW solution was 0.93 +/- 0.13 mmol/L per well per day, which was not significantly different from freshly cultured spheroids. From the aspect of urea nitrogen synthesis, spheroids cryopreserved with UW, FBS, and MIX solution were not significantly different from freshly cultured spheroids. The amount of albumin secretion by the UW cryopreserved spheroids was significantly greater than that of other cryopreserved spheroids. Cryopreserved hepatocyte spheroids in UW solution were not significantly different from freshly cultured spheroids with respect to viability and function. UW solution was superior to other cryopreservation solutions for viability and functions.

Sustained release of vascular endothelial growth factor from calcium-induced alginate hydrogels reinforced by heparin and chitosan.

A possible alternative for immunosuppression is a microencapsulation technique using hydrogels, which have been utilized for cell immobilization and drug delivery systems. Angiogenesis is crucial for delivery of the metabolic products to the host tissues as well as to supply oxygen and nutrients to cells. The local delivery of angiogenic growth factors, such as VEGF and basic FGF, has been recently studied to enhance angiogenesis on peripheral tissue of graft. In this study, we evaluated sustained VEGF release with a model using hydrogels coated with chitosan and heparin in vitro. We fabricated calcium alginate gels and chitosan-coated calcium alginate gels. Heparinized chitosan-coated calcium-induced alginate hydrogel beads were prepared by soaking chitosan-coated calcium alginate gels in heparin solution. We compared the stability and VEGF release manner between three kinds of hydrogels. To compare the stability, 5 mL of each hydrogel was incubated with 20 mL PBS under the rotational culture. Compression forces were measured using a rheometer. The amount of VEGF released from the gels was measured by ELISA. The heparin-coated chitosan alginate hydrogels showed the highest surface stability among the three hydrogels. VEGF from the heparinized gel was released in sustained manner up to 10 days in vitro. Chitosan-coated alginate gels released 90% of loaded VEGF within 5 days. These results suggest that local delivery of VEGF using a heparinized hydrogel may provide a long-term supply of angiogenic growth factor that might induce new vessel formation in vivo.

The effects of intrauterine cocaine exposure on the respiratory status of the very low birth weight infant.

OBJECTIVES: This study examined the effects of intrauterine cocaine exposure on very low birth weight infants with respect to their surfactant requirement and need for ventilatory support. STUDY DESIGN: A retrospective cohort study was conducted on infants with birth weight between 750 and 1500 g admitted to the neonatal intensive care unit between January 1992 and January 1995. RESULTS: Of the 149 infants studied, 48 infants were exposed only to cocaine and 101 infants had no drug exposure. There were no significant differences between the two groups for gestational age, sex, abruptio placenta, prolonged rupture of membranes, and antenatal steroid usage. The cocaine-exposed group had a significantly greater birth weight (1190 vs. 1109, p<0.02), less prenatal care (48% vs. 14%, p<0.00007), older maternal age (30 vs. 24, p<0.00002), more black race (79% vs. 57%, p<0.01), and more rapid plasma reagin (RPR) positivity (25% vs. 2%, p<0.00006). There were no significant differences in median APGAR scores, or incidence of necrotizing enterocolitis, retinopathy of prematurity (ROP), intraventricular hemorrhage (IVH), and bronchopulmonary dysplasia (BPD) between the two groups. Cocaine-exposed infants received surfactant treatment less often (73% vs. 48%, p<0.0035), received fewer mean doses of surfactant (0.4 vs. 10.0, p<0.0014), and were intubated less frequently (44% vs. 65%, p<0.012). There was no significant difference between groups for intubation at 24 and 48 hours and for the development of bronchopulmonary dysplasia. CONCLUSION: Perinatal cocaine exposure appears to have some significant short-term effects on the need for surfactant replacement therapy and need for initial intubation in respiratory distress syndrome (RDS) but no overall effect on the development of BPD.

A physiological role for oxalic acid biosynthesis in the wood-rotting basidiomycete Fomitopsis palustris.

A metabolic mechanism for oxalic acid biosynthesis in the wood-rotting basidiomycete Fomitopsis palustris has been proposed on the basis of biochemical analyses of glucose metabolism. There was a strong correlation between glucose consumption and oxalate production. Oxalic acid was found to accumulate in the culture fluid in about 80% of the theoretical yield or about 5-fold, on the basis of the fungal biomass harvested. The results clearly indicate that glucose was not completely oxidized to CO(2) by the tricarboxylic acid (TCA) cycle but converted mainly to oxalate. The determination of the 12 enzymes concerned has revealed the occurrence of the unprecedented metabolic coupling of the TCA and glyoxylate cycles that support oxalate biosynthesis. In this metabolic system, isocitrate lyase (EC ), together with oxaloacetase (EC ), was found to play a pivotal role in yielding oxalate from oxaloacetate via the acetate-recycling routes. Moreover, malate dehydrogenase (EC ), with an extraordinarily high activity among the enzymes tested, was shown to play an important role in generating NADH by oxidation of malate to oxaloacetate. Thus, it is proposed that the wood-rotting basidiomycete acquires biochemical energy by oxidizing glucose to oxalate.

Degradation behaviors of biodegradable macroporous scaffolds prepared by gas foaming of effervescent salts.

Biodegradable polymeric scaffolds for tissue engineering were fabricated by a gas-foaming/salt-leaching method using a combination of two effervescent salts, ammonium bicarbonate and citric acid. Poly(D,L-lactic-co-glycolic acid) (PLGA) in a state of gel-like paste was first produced by precipitation of PLGA dissolved in chloroform into ethanol. The polymer slurry was mixed with sieved particles of ammonium bicarbonate, molded, and then immersed in an aqueous solution of citric acid to generate macroporous scaffolds. The scaffolds had relatively homogeneous pore structures throughout the matrix and showed an average pore size of 200 microm and over 90% porosity. By adjusting the concentration of citric acid in the aqueous medium, it was possible to control porosity as well as mechanical strength of the scaffolds. In vitro degradation studies of three different scaffolds having lactic/glycolic acid molar ratios of 75/25, 65/35, and 50/50 exhibited marked swelling behaviors at different critical time points. The swollen matrices had a hydrogel-like internal structure. It was found that massive water uptake into the degrading scaffolds induced matrix swelling, which facilitated the hydrolytic scission of PLGA chains with concomitant disintegration of the matrices.

A novel fabrication method of macroporous biodegradable polymer scaffolds using gas foaming salt as a porogen additive.

Highly open porous biodegradable poly(L-lactic acid) ¿PLLA scaffolds for tissue regeneration were fabricated by using ammonium bicarbonate as an efficient gas foaming agent as well as a particulate porogen salt. A binary mixture of PLLA-solvent gel containing dispersed ammonium bicarbonate salt particles, which became a paste state, was cast in a mold and subsequently immersed in a hot water solution to permit the evolution of ammonia and carbon dioxide within the solidifying polymer matrix. This resulted in the expansion of pores within the polymer matrix to a great extent, leading to well interconnected macroporous scaffolds having mean pore diameters of around 300-400 microm, ideal for high-density cell seeding. Rat hepatocytes seeded into the scaffolds exhibited about 95% seeding efficiency and up to 40% viability at 1 day after the seeding. The novelty of this new method is that the PLLA paste containing ammonium bicarbonate salt particles can be easily handled and molded into any shape, allowing for fabricating a wide range of temporal tissue scaffolds requiring a specific shape and geometry.

Adhesion behaviours of hepatocytes cultured onto biodegradable polymer surface modified by alkali hydrolysis process.

Rat hepatocytes were cultured onto the surface of various amorphous biodegradable polymers composed of lactic acid and glycolic acid which were partially surface hydrolyzed by treating with sodium hydroxide. The polymer surface progressively became more hydrophilic with increasing NaOH treatment time, which was confirmed by measuring water contact angles and XPS results. The number of hepatocytes attached onto the NaOH treated hydrophilic surfaces was greater than that of the non-treated control surface. The extent of hepatocytes adhesion onto the surface-modified poly(D,L-lactic-co-glycolic acid, 85/15) depended on the NaOH treatment time. Under the optimal conditions of lactic/glycolic composition in the polymer and the surface hydrolysis time, the adhesion of hepatocytes were comparable or even better than the collagen-coated biodegradable polymer surface used as a control.

A cluster of transfusion-associated babesiosis cases traced to a single asymptomatic donor.

CONTEXT: The risk of acquiring babesiosis by blood transfusion is largely unknown since in areas where it is endemic it is often an asymptomatic infection. OBJECTIVE: To investigate and treat a cluster of blood transfusion-associated babesiosis cases. DESIGN: Case series and epidemiologic investigation. SETTING: Urban inner-city hospital. PATIENTS: Six persons who received Babesia microti-infected blood components from a donor. MAIN OUTCOME MEASURE: Diagnosis and successful therapy of babesiosis following transfusion. RESULTS: Six individuals (1 adult, 1 child, and 4 neonates) were exposed to products from a single blood donation by an asymptomatic Babesia-infected donor. Three of the 6 exposed patients became parasitemic. Polymerase chain reaction testing, animal inoculation studies, and indirect immunofluorescent antibody testing were used to confirm the presence of Babesia microti in the donor's blood and to establish the presence of infection in 3 of the 6 recipients. The 3 infected recipients and 1 additional recipient were treated without incident. CONCLUSION: Physicians should consider babesiosis in the differential diagnosis of a febrile hemolytic disorder after blood transfusion. Prompt diagnosis is important since babesiosis is responsive to antibiotic therapy and, untreated, can be a fatal disease in certain risk groups.

A comparison of meconium, maternal urine and neonatal urine for detection of maternal drug use during pregnancy.

A large scale drug screening study was done to determine the prevalence of drug use in a large metropolitan, obstetric population. Meconium and first voided urine, as well as maternal urine were collected from 423 consecutive deliveries. Urine samples and methanolic extracts of meconium were initially screened by Enzyme Multiplied Immunoassay Technique (EMIT) and then confirmed by Gas Chromatography/Mass Spectrometry (GC/MS). Analysis of cocaine metabolite as benzoylecogonine, cannabinoid as carboxy-THC, codeine, morphine and methadone were included in the study. The positive rate for benzoylecgonine was virtually identical for meconium, maternal urine and neonatal urine (12%). Analysis of meconium was found to be more reliable than analysis of maternal or neonatal urine for the detection of benzoylecgonine. Meconium did not appear to offer an advantage over maternal or neonatal urine for detection of cannabinoid, codeine, morphine, or methadone.

Human immunodeficiency virus seropositivity in critically ill neonates in the south Bronx.

Cord blood was anonymously screened to determine the prevalence of human immunodeficiency virus (HIV) seropositivity in neonates admitted to the neonatal intensive care unit (NICU) at the Bronx Lebanon Hospital Center, located in the South Bronx. We speculated that factors leading to admission to the NICU such as low birth weight, prematurity and being small for gestational age would also be associated with an increased prevalence of HIV seropositivity. During the study period the prevalence of HIV seropositivity was 11.6% in the NICU population. There was no significant difference in maternal age, gravidity, race and sex in HIV-seropositive vs. HIV-seronegative newborns. There was a significantly increased incidence of maternal drug use (P less than 0.01), babies small for gestational age (P less than 0.005) and microcephaly (P less than 0.02) in seropositive vs. seronegative NICU babies. The results of this study suggest that the NICU population may comprise a significant number of infants of HIV-infected mothers.

Neuropathic arthrogryposis multiplex congenita and intrauterine ischemia of anterior horn cells: a hypothesis.

Three patients with multiple articular deformities due to arthrogryposis multiplex congenita (AMC) are presented. All had severe thalamic and tegmental degenerations with glial scars and shrunken ferruginated neurons; one had also unilateral porencephaly and polymicrogyri. The character and the pattern of the lesions were highly suggestive of in-utero hypotensive/ischemic insult. Marked attenuations of anterior spinal roots were mostly seen in the lumbosacral and cervical enlargements, where anterior horn cell loss and ferrugination were most severe. The changes in the anterior horns were situated peripherally, a distribution corresponding to the terminal ramifications of the sulcal (central) branches of the anterior spinal artery, an area known for its susceptibility to ischemia, especially in children. The changes in the brains and spinal cords could reasonably be explained on the basis of intra-uterine perfusion failure in early stages of fetal development. This assumption was supported by the fact that all three patients were subjected to unfavorable environmental factors during gestation. The first was the twin of a macerated infant; their entangled umbilical cords might have caused differential placental perfusion resulting in partial ischemia in the twin with AMC and severe ischemia in the other accounting for its early demise. The second patient had a single functional umbilical artery, a condition frequently associated with developmental anomalies. The third was born to a mother whose pregnancy was complicated by heavy substance abuse. Ischemic myelopathy during a critical phase of CNS developmental and muscle innervation is proposed as a cause of some cases of neuropathic AMC. Placental circulatory failure should be considered in the pathogenesis of AMC in such cases.

Intrapartum rupture of the falciform ligament and umbilical vein. A rare cause of hemoperitoneum in the newborn.

Intra-abdominal hemorrhage in the newborn is uncommon, but it must be considered in the first 48 hours of life in the infant with pallor, anemia, abdominal distension, and shock. The injured liver is the most common source of bleeding, with the spleen and kidney less often involved. In the case presented, the hallmarks of intra-abdominal hemorrhage were evident. Exploratory laparotomy revealed intraperitoneal bleeding emanating from the disruption of the umbilical vein and its enveloping falciform ligament. There was no other site of intra-abdominal bleeding and there were no intrinsic abnormalities of the umbilical cord or the placenta. Disruption of the intra-abdominal umbilical vein represented the sole source of intra-abdominal bleeding in this patient. The case is reported to document disruption of the intra-abdominal umbilical vein as a rare cause of neonatal hemoperitoneum.

Pulmonary candidiasis in infants: clinical, radiologic, and pathologic features.

Although systemic candidiasis is common in hospitalized patients, invasive pulmonary candidiasis is rare and generally considered of secondary importance when found at autopsy. Autopsy records for a 12 year period were reviewed and 15 infants were found in whom systemic candidiasis was considered the primary or a major contributory cause of death. Significant pulmonary involvement was found in 14. There were three characteristic histologic patterns of pulmonary candidiasis: (1) embolic (arterial-invasive) (seven cases); (2) disseminated (capillary-invasive) (four cases); and (3) bronchopulmonary (air space-invasive) (three cases, including one congenital infection). An indwelling vascular catheter or infected cutdown wound was the portal of entry in every case of the embolic form of pulmonary candidiasis. Systemic and pulmonary Candida infections were rarely diagnosed during life. The typical radiographic appearance was progressive air space consolidation, although two infants with the embolic form of pulmonary candidiasis had focal cavitation. In general, there was poor correlation between the radiologic and pathologic findings, and pathologic findings other than pulmonary candidiasis undoubtedly accounted for many of the radiologic abnormalities observed. There were no radiologic findings that could be used to differentiate the three histologic forms of lung involvement in these patients. Small lung nodules, the earliest histologic lesion of pulmonary candidiasis, were not seen in any patient owing to the presence of other lung disease and suboptimal radiographic technique.

Lack of predictive indices in kernicterus: a comparison of clinical and pathologic factors in infants with or without kernicterus.

A review of 398 neonatal autopsies at Downstate Medical Center revealed 27 cases of kernicterus during the seven-year period from 1971 through 1977. With the current intensive care of the sick newborn, kernicterus continues to occur, mainly in premature infants with relatively low levels of serum bilirubin (mean of 11.5 mg/100 ml). To understand the factors contributing to the development of kernicterus, clinical and pathologic findings in 27 infants with kernicterus were compared to 103 "control" infants with retrospectively. Birth weight, gestational age, sex, and Apgar scores were comparable in both groups. The duration of survival was significantly shorter in infants with kernicterus than in the control infants. The clinical signs and symptoms of kernicterus were nonspecific and the premortem diagnosis of kernicterus was not suspected in most of the cases. There were no significant differences in the peak serum bilirubin values, incidence of hypothermia, hypoglycemia, convulsions, anemia, infection, use of phototherapy, transfusion and exchange transfusion in the two groups. Serum albumin values and bilirubin binding capacity measured by 2-(4-hydroxybenzeneazo)benzoic acid were significantly lower in the kernicteric group although the bilirubin-albumin molar ratio was equal in both groups. The incidences of severe acidosis and hypoxic encephalopathy were significantly higher in the kernicteric infants. In this study, acidosis, hypoxia, hypoalbuminemia, and low bilirubin binding capacity were seen more often in kernicteric infants than in control infants. However, analysis of previously suggested risk factors failed to identify any single factor or combination of factors which could be predictive to the development of kernicterus.