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Biomedical subjects

L H Lee

Publications and source records attributed to L H Lee.

At least 73 records · Page 4Linked to original sources

Respiratory nuclei share synaptic connectivity with pontine reticular regions regulating REM sleep.

Injection of cholinomimetics into the medial pontine reticular formation (mPRF) of intact, unanesthetized cat causes a rapid eye movement (REM) sleep-like state and respiratory depression. The mPRF contains no concentrations of respiratory neurons, and this study examined the hypothesis that respiratory depression evoked from the mPRF is synaptically mediated. The mPRF of conscious cats was injected with bethanechol to define an mPRF zone causing state-dependent respiratory depression. Bethanechol caused a 361% increase in the REM sleep-like state and a 37% decrease in minute ventilation. Additional cats were injected with the retrograde fluorescent tracers True Blue and either Fluoro-Gold or Diamidino Yellow aimed for the cholinoceptive mPRF or for the pontine respiratory group (PRG). After mPRF dye injection, 1) labeling was observed in the PRG, dorsal respiratory group (DRG), and ventral respiratory group (VRG); and 2) double-labeled cells were observed in the VRG and PRG. Dye injections into the PRG produced contralateral and ipsilateral fluorescent labeling of the mPRF, DRG, and VRG. Thus cholinoceptive regions of the mPRF involved in REM sleep generation have reciprocal monosynaptic connections with the PRG and receive monosynaptic projections from the DRG and VRG.

Animals↗

Haemophagocytosis in bone marrow aspirate--a review of the clinical course of 10 cases.

The clinical course of 10 cases where marrow aspirate showed features of haemophagocytosis was reviewed. Eight of these had a fulminant clinical course characterized by high fever, constitutional symptoms, wasting, hepatosplenomegaly with liver dysfunction, sometimes lymphadenopathy, progressive pancytopenia and coagulopathy, like that described as 'malignant histiocytosis' in the past. The remaining 2 cases did not have this classical clinical syndrome. Among the former 8 cases, 4 of them had high-grade lymphoma, 3 of whom were confirmed to be peripheral T cell lymphoma. Three of the remaining 4 had suspicious lymphomatous infiltrate on marrow trephine. In every case an extensive search for viral etiology by serology was negative. The 2 cases which did not have fulminant clinical feature were found to have lymphoma of the diffuse large cell and Ki-1 anaplastic type, respectively. A review of the literature reveal that most cases with haemophagocytic syndrome have a fulminant clinical course and are peripheral T cell lymphoma, which generally has a poor prognosis. In our study, the 8 cases with the classical haemophagocytic syndrome had a median survival of 24 days and a long-term survival of 37.5% at 28 months. Prompt initiation of chemotherapy is a life-saving measure and the only chance of achieving a long-term survival in patients with haemophagocytic syndrome if the underlying lymphoma can be diagnosed early.

Adolescent↗

Long-term propofol infusion and airway management in a patient with Goldenhar's syndrome.

A 2-year-old patient of Goldenhar's syndrome received an operation for corneal transplantation. Difficult endotracheal intubation from the congenital anomaly was treated with laryngeal mask airway and pediatric fiberoptic laryngoscope. Long-term propofol infusion (> 10 h) for anaesthetic maintenance in this small child was used with rapid and smooth recovery.

Anesthetics, Intravenous↗

Analysis of the double-stranded RNA genome segments among avian reovirus field isolates.

Nine isolates of avian reovirus (ARV) from both healthy birds and birds with different clinical illness and one commercially available vaccine strain were selected and characterized by analysis of the migration pattern of their genomic double-stranded RNA (dsRNA) segments following separation by polyacrylamide gel electrophoresis. Different electropherotypes were observed and analyzed. The results show that the dsRNA segments of ARV were markedly polymorphic among isolates within the same serotype as well as among different serotypes. The results also show no correlation between electropherotype and disease state.

Animals↗

Comparison of the labeled avidin-biotin and the conventional enzyme-linked immunosorbent assay for detecting antibody to reovirus in chickens.

A labeled avidin-biotin enzyme-linked immunosorbent assay (LAB-ELISA) for detecting antibodies to avian reovirus (ARV) in chicken sera was developed and compared with conventional ELISA. Purified ARV, biotin-labeled rabbit anti-chicken IgG conjugate, and horseradish peroxidase-labeled avidin were used in the LAB-ELISA. The two types of ELISA had the similar ability to clearly distinguish ARV-positive and -negative sera. Furthermore, non-specific reactions with the two ELISAs against sera from ARV-negative chickens were reduced markedly. When sera from farm chickens were tested by the two ELISAs and serum neutralization (SN), the correlation rates between SN and conventional ELISA, and SN and LAB-ELISA were similar and were 93.4% and 92.3%, respectively.

Animals↗

Heat shock- and alkaline pH-induced proteins of Campylobacter jejuni: characterization and immunological properties.

The protein response to physiological stress was characterized in Campylobacter jejuni 81176 after exposure to heat and pH shock and following periods of recovery. Immunoreactivities of major stress-related proteins were determined with anti-Campylobacter immune rabbit serum and intestinal lavage fluid. Distinct proteins with molecular masses ranging from 10 to 120 kDa were induced and/or released by selective heat or pH treatments. The most notable responses were those of two proteins with apparent molecular masses of 45 and 64 kDa that were induced and two other proteins of 10 and 12 kDa that were released by selective heat shock, alkaline pH treatment, or both. On the basis of N-terminal sequence analysis and immunological cross-reactivity data, the 64- and 10-kDa proteins were the C. jejuni homologs of Escherichia coli GroEL and GroES proteins, respectively. Enhanced chemiluminescence Western blotting (immunoblotting) revealed that all four proteins were among the major protein antigens recognized by anti-Campylobacter rabbit serum immunoglobulin G (IgG) and immune rabbit intestinal lavage IgA (secretory IgA). The results of this investigation suggest that the C. jejuni 10-, 12-, 45-, and 64-kDa proteins and a number of minor stress-related proteins deserve further evaluation of their respective roles in Campylobacter pathogenesis and immunity.

Amino Acid Sequence↗

Single-tube, noninterrupted reverse transcription-PCR for detection of infectious bursal disease virus.

An assay protocol based on single-tube, noninterrupted reverse transcription-PCR (RT-PCR) for the detection of infectious bursal disease virus (IBDV) is described. After the conditions for RT-PCR had been optimized, a primer set framing a region within the gene coding for IBDV VP2 protein was used to amplify a 318-bp fragment of the IBDV genome. Amplified product was detected with three strains of IBDV, whereas none was obtained from uninfected bursal tissue or seven unrelated avian viruses. The sensitivity of this RT-PCR was tested with purified viral RNA from three strains of IBDV. The detection limit was 10 fg in an ethidium bromide-stained gel. In addition, this assay system was used to detect IBDV in bursal-tissue specimens from commercially reared chickens. The identity of the amplified products from the tissue specimen preparation was determined by using a rapid, simple procedure in which internally nested, end-labeled probes were used.

Animals↗

Markers of thrombin and plasmin generation in patients with inherited thrombophilia.

AIM: To determine the prevalence of a biochemically detectable hypercoagulable state, defined in terms of increased thrombin or plasmin generation, in patients with phenotypically characterised thrombophilia. METHODS: Plasma concentrations of the prothrombin activation peptide F1.2 and fibrin degradation (FbDP) and fibrinogen degradation products (FgDP) were measured by enzyme immunoassay in 104 patients deficient in natural anticoagulants, and 35 unaffected relatives. RESULTS: Increased concentrations of F1.2, FbDP, and FgDP were present in 18, 25, and 19 of 104 patients, respectively. There were no correlations between F1.2, FbDP, and FgDP concentrations, or between these parameters and concentrations of natural anticoagulants except for a negative correlation between protein C concentrations and FgDP (rho = -0.46, p = 0.009). CONCLUSION: A biochemically detectable hypercoagulable state is present in some patients with asymptomatic thrombophilia. Markers of plasmin generation may be increased more frequently in thrombophilia than markers of thrombin generation. This finding should prompt the inclusion of markers of plasmin generation in prospective longitudinal cohort studies to determine the predictive value of a hypercoagulable state, defined by either excessive thrombin or plasmin generation, for the development of venous thromboembolism.

Adult↗

Haemostasis associated with liver disease and liver transplantation.

The production and regulation of components of haemostasis is largely dependent on liver function. Liver disease is associated with alteration of the coagulant:anticoagulant and fibrinolytic:antifibrinolytic balance as well as quantitative and qualitative alterations of the cellular components of haemostasis. In this article we have reviewed the nature and frequency of disturbed haemostatic function in patients with acute and chronic liver disease and during liver transplantation. The risk of haemorrhage and thrombosis, and mechanisms leading to these complications are reviewed and treatment strategies utilising blood component therapy and pharmacological manipulation of the coagulant and fibrinolytic systems are proposed. Finally, areas for further study are suggested so that rational effective therapeutic strategies can be developed.

Acute Disease↗

Detection of infectious bursal disease virus infection using the polymerase chain reaction.

The method of reverse transcription (RT) followed by the polymerase chain reaction (PCR) was used to amplify two different fragments of the infectious bursal disease virus (IBDV) genome. Two sets of primer framed two different regions within the genes coding for proteins VP2 and VP3, respectively. Both sequences were detected in five strains of IBDV, whereas, none were obtained from uninfected control cells. The sensitivity of RT-PCR was carried out on nucleic acids from the IBDV infected cell cultures. The detection limit was 10(0) to 10(-1) TCID50 in ethidium bromide stained gels and could be enhanced further to 10(-1) to 10(-3) TCID50 by hybridization after southern transfer. In addition, detection of IBDV infection in 12 out of 14 bursal specimens examined by this technique was shown to be entirely consistent with the clinical history and an alternative diagnostic method. The speed, sensitivity, and specificity of this method is relevant for the diagnosis of infection with IBDV.

Animals↗

Long-term survival of mouse corpus callosum grafts in neonatal rat recipients, and the effect of host sensitization.

Previous studies have suggested that the incidence of spontaneous rejection among immunogenetically mismatched neural transplants in neonatal recipients varies significantly depending on the cellular composition of the graft material. For example, neuron-rich grafts of embryonic mouse retina generally survive for extended periods without showing signs of rejection after implantation into neonatal rats, whereas cortical xenografts, which contain abundant glial and endothelial cells as well as neurons, typically undergo rejection 4-6 weeks after implantation. To determine whether the presence of donor glia is responsible for this high incidence of spontaneous rejection, we examined the fate of a non-neuronal graft material composed predominantly of xenogeneic glial cells (post-natal day 3, PD3, CD-1 mouse corpus callosum) implanted into the mesencephalon of PD1 Sprague-Dawley rats. The distribution and survival of donor astrocytes were assessed using a monoclonal antibody specific for a mouse astrocyte surface antigen, M2. Thirteen of 16 animals sacrificed within 2 months of implantation had detectable transplants. In these animals, M2-positive cells frequently migrated well away from body of the graft, clustering in large numbers in several characteristic regions of the host brain. Unlike cortical grafts of similar age, the vast majority (93%) of callosal transplants showed no histological signs of rejection or major histocompatibility complex antigen expression in and around the transplant-derived cells. As previously noted in the neonatal retinal transplant paradigm, however, well-integrated 1-month-old corpus callosum grafts could be induced to reject by appropriate sensitization of the host immune system, implying that the host was not immunologically tolerant to the foreign neural graft. With longer survival times in unsensitized hosts, a progressively smaller percentage of animals had detectable donor astrocytes (5 of 10 animals at 3 months postimplantation and 4 of 16 animals at 4 months); in those 9 animals with surviving grafts, only small numbers of M2-positive cells were seen within the graft bed and surrounding host brain. However, only 2 of the 26 "long-term" animals showed evidence of graft rejection. These results indicate that mouse astrocytes show characteristic patterns of migration into the host brain when implanted into neonatal rats; however, these xenogeneic cells have a limited duration of survival. The infrequency with which even subtle signs of spontaneous rejection were detected in animals that had received corpus callosum xenografts suggests that an immune-mediated process is unlikely to be responsible for the time-dependent elimination of the donor astrocytes.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

A monoclonal antibody capture enzyme-linked immunosorbent assay for detecting antibodies to infectious bursal disease virus.

A monoclonal antibody capture enzyme-linked immunosorbent assay (mAb-ELISA) for antibodies to infectious bursal disease virus (IBDV) in chicken sera was developed and compared with conventional ELISA. When sera from farm chickens were tested by the two ELISAs and serum neutralization (SN), the correlation rate between SN and mAb-ELISA was 100% (49/49), and that between SN and conventional ELISA was 81.6% (40/49). In mAb-ELISA, all of the sera that were antibody-negative by SN had low absorbance values (below 0.05), and the absorbance values correlated closely with the SN titers. In the conventional ELISA, however, the sera antibody-negative by SN had various absorbance values ranging from 0.06 to 0.32. mAb-ELISA had much lower non-specific reactions than the conventional ELISA against sera from IBD-negative chickens.

Animals↗

Characterization of nonradioactive hybridization probes for detecting infectious bursal disease virus.

Reverse transcription followed by the polymerase chain reaction was used to amplify a fragment of infectious bursal disease virus (IBDV) strain P3009 genome. The amplified DNA fragment was annealed into the plasmid pUC18 and used to transform Escherichia coli strain JM109. A clone that contained IBDV-specific nucleotide sequences was selected and designated pC23. The DNA fragment within pC23 was 320 base pairs in length and designated C23. Radiolabeled probes prepared from C23 hybridized to genome segment A of strain P3009 by a northern-blot hybridization assay. Biotin-labeled probes prepared from C23 and pC23 either by using nick translation (designated C23/NT and pC23/NT, respectively) or by direct introduction of biotin molecules into C23 and pC32 (designated C23/BH and pC23/BH, respectively) were used in the dot blot hybridization assay for detecting IBDV strains. All four biotinylated probes detected three serotype 1 viruses and one serotype 2 IBDV. However, they did not cross-react with nucleic acids extracted from mock-infected cells or from seven unrelated avian viruses. Probe pC23/BH detected as little as 0.04 ng of IBDV RNA, while the other three probes were less sensitive and detected approximately 1 ng of IBDV RNA. In addition, the probe pC23/BH detected IBDV RNA in bursa tissues from commercial broiler raising farms following the dot blot hybridization.

Animals↗

Acute toxicity, distribution, and metabolism of 2,4,6-trinitrophenol (picric acid) in Fischer 344 rats.

Picric acid (2,4,6-trinitrophenol) is widely used in industry, by the military, and as a research/clinical chemistry reagent. Characterization of the toxicity of this chemical has been limited. Thus the acute toxicity, distribution, and metabolism of picric acid were investigated using Fischer 344 rats. The LD50 for picric acid following oral dosing of male and female rats was established as 290 and 200 mg/kg, respectively. Blood gas analysis indicated severe acidosis during acute intoxication. Metabolism of picric acid was limited to reduction of nitro groups to amines. Metabolites isolated from urine included N-acetylisopicramic acid (14.8%), picramic acid (18.5%), N-acetylpicramic acid (4.7%), and unidentified components (2.4%). Approximately 60% of the parent picric acid was excreted unchanged. The plasma half-life for picric acid was 13.4 h with a gut absorption coefficient (ka) of 0.069 h-1. Twenty-four hours following oral administration of [14C]picric acid (100 mg/kg), the primary depots of radioactivity (per gram tissue basis) were blood, spleen, kidney, liver, lung, and testes. Respective tissue/blood ratios were 0.37, 0.31, 0.28, 0.26, and 0.22. All other tissue assayed had partition ratios < 0.20, with brain and adipose tissue having the least amount of radioactivity. Tissue/blood ratios were essentially maintained over a 48-h postadministration period. Binding (in vitro) of [14C]picric acid to plasma proteins (whole blood preparations) demonstrated both high- and low-affinity binding sites, with dissociation constants of 3.18 x 10(-6) and 2.85 x 10(-4) M, respectively. The findings of this investigation provide information on the acute toxicity, metabolism, and distribution of picric acid, which can be used in risk assessment analyses and in the design of subchronic and chronic toxicity tests.

Acidosis↗