Biomedical subjects

K H Nguyen

Publications and source records attributed to K H Nguyen.

At least 19 recordsLinked to original sources

PubMed2000

Stereospecific versus nonstereospecific assessments for the bioequivalence of two formulations of racemic chlorpheniramine.

Chlorpheniramine (chlorphenamine, CPAM) is a racemic antihistaminic H1 drug containing two enantiomers. The aim of this study was to assess the bioequivalence of two formulations (reference and Vietnamese-tested formulation) of racemic chlorpheniramine combined with phenylpropanolamine in an open-labeled, randomized, crossover two-period study, after administration of 8 mg of racemic chlorpheniramine in 12 healthy Vietnamese subjects. First, dissolution of both formulations was tested in vitro according to USP requirements. Then the 12 subjects received both formulations after an overnight fast and a 7-day wash-out period. Plasma samples were collected up to 168 h. Plasma concentrations of total chlorpheniramine and its individual enantiomers were determined with a validated chiral HPLC method and pharmacokinetic parameters were estimated using model-independent analysis. For the reference formulation, Cmax and AUC values were higher for (+)S-chlorpheniramine ((+)S-CPAM) compared to (-)R-chlorpheniramine ((-)R-CPAM) (13.3 vs. 6.8 ng/ml and 409 vs. 222 ng/ml/h, respectively) while Clt/F and Vd/F were lower (9.8 vs. 17.6 l/h and 321 vs. 627 l, respectively). No difference was observed for Tmax, t(1/2), and MRT. Pharmacokinetic parameters were similar for the reference and the Vietnamese-tested formulation. Bioequivalence was assessed by Schuirmann test, as recommended by the current FDA and European Community criteria. Dissolution tests showed that both formulations were equivalent. A nonstereospecific, but not a stereospecific, approach indicated bioequivalence between the formulations.

Adult↗

PubMed2000

Evidence for antimuscarinic acetylcholine receptor antibody-mediated secretory dysfunction in nod mice.

OBJECTIVE: Antibodies directed against general and specific target-organ autoantigens are present in the sera of human patients and animal models with autoimmune disease. The relevance of these autoantibodies to the disease process remains ambiguous in most cases. In autoimmune exocrinopathy (Sjögren's syndrome), autoantibodies to the intracellular nuclear proteins SSA/Ro and SSB/La, as well as the cell surface muscarinic cholinergic receptor (M3) are observed. To evaluate the potential role of these factors in the loss of secretory function of exocrine tissues, a panel of monoclonal and polyclonal antibodies was developed for passive transfer into the NOD animal model. METHODS: Monoclonal antibodies to mouse SSB/La, rat M3 receptor, and a rabbit polyclonal antiparotid secretory protein antibody were obtained for this study. These antibody reagents were subsequently infused into NOD-scid mice. Saliva flow rates were subsequently monitored over a 72-hour period. Submandibular gland lysates were examined by Western blotting for alteration of the distribution of the water channel protein aquaporin (AQP). RESULTS: Evaluation of the secretory response indicated that only antibodies directed toward the extracellular domains of the M3 receptor were capable of mediating the exocrine dysfunction aspect of the clinical pathology of the autoimmune disease. In vitro stimulation with a muscarinic agonist of submandibular gland cells isolated from mice treated with anti-M3 antibody, but not saline or the isotype control, failed to translocate AQP to the plasma membrane. CONCLUSION: These findings define a clear role for the humoral immune response and the targeting of the cell surface M3 signal transduction receptor as primary events in the development of clinical symptoms of autoimmune exocrinopathy. Furthermore, the anti-M3 receptor activity may negatively affect the secretory response through perturbation of normal signal transduction events, leading to translocation of the epithelial cell water channel.

Animals↗

PubMed2000

Congenital Heart Surgery Nomenclature and Database Project: aortic valve disease.

The extant nomenclature for aortic valve disease is reviewed for the purpose of establishing a unified reporting system. The subject was debated and reviewed by members of the STS-Congenital Heart Surgery Database Committee and representatives from the European Association for Cardiothoracic Surgery. All efforts were made to include all relevant nomenclature categories using synonyms where appropriate. Aortic valve disease has been subdivided into stenotic and regurgitant lesions. Stenotic lesions have been characterized by anatomic location: supravalvar, valvar, and subvalvar. Regurgitant lesions have been characterized as either congenital or acquired. A comprehensive database set is presented that is based on a hierarchical scheme. Data are entered at various levels of complexity and detail that can be determined by the clinician. These data can lay the foundation for comprehensive risk stratification analyses. A minimum database set is also presented that will allow for data sharing and would lend itself to basic interpretation of trends. Outcome tables relating diagnoses, procedures, and various risk factors are presented.

Aortic Valve↗

PubMed2000

Salivary epidermal growth factor levels decrease in patients receiving radiation therapy to the head and neck.

OBJECTIVE: The objective of this study was to assess changes in salivary epidermal growth factor (EGF) in patients receiving radiation therapy to the head and neck and to determine whether salivary EGF levels correlate with the severity of radiation-induced oral mucositis. STUDY DESIGN: Thirteen patients and 18 control subjects were enrolled in the study. Saliva was collected before, during (weekly), and after radiation therapy. Salivary total protein (TP) and EGF concentrations were measured and correlated with the severity of oral mucositis. The variability in normalized EGF (ngEGF/mgTP) values and mucositis scores were analyzed with analysis of covariance, and the adjusted correlation coefficient was calculated. RESULTS: EGF levels decreased (P =.004), whereas TP levels increased over time (P =.039). A strong correlation was seen with decreasing normalized EGF values and more severe mucositis (P =. 0001). CONCLUSION: A strong negative correlation between normalized EGF and mucositis severity suggests a possible role for EGF in the progression of radiation-induced mucosal breakdown.

Adult↗

PubMed1999

Subtypes of acute aortic dissection.

BACKGROUND: This series consists of a 12-year experience with a policy of identifying and replacing the aortic segment containing the primary intimal tear for repair of acute aortic dissection. METHODS: Patients with type A dissection underwent urgent surgery. Patients with type B dissection were referred for surgery based on selective criteria, including aortic dilatation greater than 5 cm. A classification system for acute dissection is described that specifies the site of intimal tear while retaining the clinical relevance of the Stanford system. RESULTS: Of 168 acute dissections, 139 were type A and 29 were type B. The site of intimal tear was as follows: ascending aorta, 83 cases; arch, 32 cases; descending aorta, 29 cases; multiple tears, 11 cases (10 included arch tears); no tear (intramural hematoma), 6 cases; not noted, 7 cases. Only 60% of acute type A dissections arose from solitary intimal tears in the ascending aorta, whereas 30% had arch tears. Hospital mortality for type A dissection was 13.7% (18.8% for arch tears, NS) and 0% for type B. False lumen patency was 57.1% for type A dissection and 18.8% for type B dissection (p = 0.002), yet survival was similar for these groups. Ten-year survival for type A dissection with arch tear (0.51 +/- 0.12) was lower than 10-year survival for type A dissection with ascending tear (0.74 +/- 0.05; p = 0.77), and significantly lower than for type A dissection with descending tear (0.88 +/- 0.12; p = 0.029). CONCLUSIONS: Systematic resection of the primary tear yielded similar hospital mortality, 5-year survival, and aorta-related event-free survival rates for subtypes of acute type A dissection. Excellent results were obtained with a selective approach to type B dissection.

Acute Disease↗

PubMed1999

Natural history of descending thoracic and thoracoabdominal aneurysms.

BACKGROUND: A review of 165 patients with chronic dissecting and degenerative aneurysms of the descending thoracic and thoracoabdominal aorta initially managed nonoperatively was carried out to ascertain factors associated with a high risk of rupture. METHODS: Changes in the aneurysms were followed with three-dimensional reconstructions of computed tomograph scans. Risk factors were compared in patients with dissecting and nondissecting aneurysms who experienced rupture, in whom operation was recommended during the course of follow-up, and in those without rupture or operation. RESULTS: Nondimensional variables associated with an enhanced risk of rupture include age, the presence of chronic obstructive pulmonary disease, and even uncharacteristic continued pain. Patients with rupture of dissections had significantly higher blood pressures than survivors, and significantly smaller maximal descending thoracic aortic diameters (median 5.4 cm) than patients with rupture of degenerative aneurysms (median 5.8 cm). The extent of the aneurysm, as reflected by the maximal abdominal aortic diameter, was a significant risk factor for rupture only in nondissecting aneurysms. Mortality from rupture was significantly higher in patients with chronic dissections than in patients with nondissecting aneurysms: 9/10 vs 26/34 (p = 0.004). CONCLUSIONS: Almost 20% of patients followed nonoperatively succumbed to rupture, suggesting that a more aggressive surgical approach toward patients with chronic aneurysms of the descending thoracic and thoracoabdominal aorta is warranted. An individualized risk of rupture within 1 year can now be calculated, and patients whose operative risk is lower than their calculated risk should be offered elective surgery.

Aortic Dissection↗

PubMed1999

Use of somatosensory evoked potentials for thoracic and thoracoabdominal aortic resections.

BACKGROUND: Despite tremendous development in surgical and anesthetic techniques, resection of the thoracic and thoracoabdominal segments of the aorta remain associated with the risk of paralysis. Routine use of somatosensory-evoked potential (SEP) monitoring in patients undergoing surgery of the thoracic aorta has become a standard intra- and postoperative procedure at our institution since its first use in 1993. METHODS: One hundred forty nine (149) thoracic aortic operations were performed during January 1993 through January 1998 using SEP-directed serial sacrifice of paired intercostal arteries. Full, partial, or no cardiovascular bypass was variably used, dictated by anatomy; 49 patients required deep hypothermic circulatory arrest (DHCA). Patients were monitored during both the intraoperative procedure as well for the post-anesthesia period until neurologic stability and/or ability to reproducibly demonstrate lower extremity neurologic competency was established. Postoperative neurologic function was compared to ischemic intervals, extent of aortic resection, number of intercostal arteries sacrificed, type of perfusion, and underlying aortic pathology. RESULTS: Overall mortality in the group was 13 patients (8.7%), with no one cause predominating. Nine patients sustained permanent paraplegia, only 1 of whom lost SEPs during the procedure. Abnormal SEPs were seen in 19 patients, 14 of whom had normal neurologic function after awakening. Three of 19 (15.8%) developed late paraplegia that resolved with medical therapy. Eleven patients (7.4%) developed cerebrovascular accidents (CVA), with the majority (8) appearing in the group undergoing DHCA. The risk of CVA was significantly higher in DHCA patients (p < 0.01) than other patients. No patient with CVA had abnormal SEPs; 4 DHCA patients developed abnormal SEPs, 1 with permanent paralysis. CONCLUSIONS: The routine use of SEP monitoring during thoracic and thoracoabdominal aortic surgery as well as during the postoperative period may be useful in decreasing the observed incidence of paraplegic events associated with these procedures.

Adolescent↗

PubMed1999

Intra-articular IL-4 gene therapy in arthritis: anti-inflammatory effect and enhanced th2activity.

Gene therapy has been explored as a potential method for treating chronic inflammatory diseases such as rheumatoid arthritis. To determine the efficacy of intra-articular IL-4 gene therapy in an animal model of arthritis using a retroviral vector, a retrovirus encoding rat IL-4 (DA-IL-4) was engineered, purified and concentrated to high titer (>/=109 CFU/ml). Infectivity and expression levels were demonstrated in vitro using cultured fibroblast-like synoviocytes. Efficacy was evaluated in the rat adjuvant arthritis model. DA-IL-4 or DA-beta-gal retrovirus was injected into the intra-articular joint space of the right ankle on day 12 after immunization. Three days after joint injection, the injected paw contained increased levels of IL-4 compared with control or with the contralateral uninjected paw, demonstrating successful transgene expression. Surprisingly, 8 days after treatment IL-4 levels continued to increase in the injected and contralateral paw compared with DA-beta-gal-treated animals. Serum IL-4 levels were also elevated in DA-IL-4-treated rats. RT-PCR studies demonstrated that the transgene was expressed in the injected ankle but not in the contralateral joint. IL-4 gene therapy resulted in a significant reduction in paw swelling and decreased radiographic evidence of bone destruction. This is the first demonstration of successful intra-articular retroviral gene treatment using a therapeutic gene. In addition to its anti-inflammatory effect, this study supports the potential application of intra-articular gene therapy as a method for enhancing systemic Th2 function.

Animals↗

PubMed1998

The outcome of cardiac operations in infants weighing two kilograms or less.

OBJECTIVE: A review of our recent experience of operating on infants weighing 2 kg or less who had congenital heart disease was performed to determine the outcome of early surgical repair or palliation. METHODS: A retrospective review of hospital records was performed for infants who weighed 2 kg or less and who were identified to have undergone cardiac operation at our institution January 1992 to June 1997. The data collected included age, weight, gestational age, cardiac diagnosis, surgical procedure, and outcome measures such as length of stay, morbidity, and mortality rate. Outpatient charts were reviewed for follow-up survival and cardiac status. RESULTS: Thirty-three operations were performed on 30 patients. Median age at operation was 19.5 days (1 to 140 days), and median weight was 1.8 kg (1.1 to 2.0 kg). Cardiac diagnoses varied, with coarctation of the aorta and tetralogy of Fallot most common. Twenty-four patients were born at 37 or fewer weeks' gestation. Hospital survival was 83% with no difference in mortality rates based on age, weight, or type of surgical procedure. Premature infants tended to have worse hospital survival. Median postoperative length of stay was 39 days (6 to 122 days). Median duration of mechanical ventilation in survivors was 6 days (2 to 24 days). Neurologic complications were documented in eight patients. Of the 25 hospital survivors, 20 (80%) are alive with good cardiac status at a mean follow-up of 13 months. CONCLUSION: Cardiac operations in a selected group of infants weighing 2 kg or less can provide acceptable hospital survival. In most instances, complete repair is possible with good medium-term outcome in the survivors. Investigation into neurologic outcomes in these patients is warranted.

Cardiac Surgical Procedures↗

PubMed1998

Changes in the expression of extracellular matrix (ECM) and matrix metalloproteinases (MMP) of proliferating rat parotid acinar cells.

Tissue morphogenesis, development, and maintenance of function are mediated by signals generated through the composition of the extracellular matrix. The regulation of the composition of matrix is determined by enzymes specific for their degradation, the matrix metalloproteinases. Chronic injections of the beta-adrenergic receptor agonist, isoproterenol, result in a non-neoplastic hypertrophy and hyperplasia of the rat parotid gland. The activity of matrix metalloproteinases, as measured by gelatin zymography and enzymatic digestion of Azocoll substrates by gland lysates, decreased significantly (P < 0.05) following 24 hrs of agonist treatment, and slowly recovered to control values by 6 days of treatment. Daily administration of the broad-spectrum matrix metalloproteinase inhibitor Galardin for 3 days in combination with isoproterenol resulted in enhanced gland hypertrophy compared with that produced by isoproterenol alone. Given alone, Galardin also caused hypertrophy. The relative abundance of mRNA for the extracellular matrix molecules, collagens I and III and fibronectin, declined rapidly following the initiation of beta-agonist treatment in vivo, while laminin B1 and B2 mRNA levels increased initially before declining below control levels. These changes in patterns of mRNA levels also were observed in the concentrations of glandular protein when Western dot blot analysis of collagens I and III and laminin, respectively, was used. The importance of laminin, in vivo, was demonstrated by coinjection of anti-laminin antibody along with isoproterenol, which resulted in the inhibition of beta-agonist-induced parotid gland hypertrophy and hyperplasia. These data suggest that modulation of the ECM is associated with isoproterenol-induced salivary gland hypertrophy and hyperplasia. It is likely that this modulation of the ECM takes place through transcriptional regulation of some ECM genes and regulation of matrix-degrading enzyme activity.

Adrenergic beta-Agonists↗

PubMed1998

Elevated salivary EGF levels stimulated by periodontal surgery.

Previous studies have shown epidermal growth factor (EGF) to be involved in oral wound healing as well as gastric cytoprotection. EGF functions with hormone-like properties to stimulate epithelial cells by binding to the EGF receptor which ultimately leads to proliferation via signal transduction mechanisms. Salivary glands are a major source of EGF The purpose of this study was to determine if intra-oral wounding by periodontal surgery stimulated increased salivary EGF levels. Salivary EGF levels were determined for 12 systemically healthy individuals (ages 27 to 70 years old) presurgically and postsurgically at 6, 12, 18, 24, 30, 36, and 42 hours and 2 and 6 weeks. Three ml of unstimulated whole saliva was obtained at each time point to allow immunoassay quantitation. Age and gender matched unoperated patients served as controls. Salivary samples were incubated with monoclonal and polyclonal EGF antibodies in these "sandwich" enzyme immunoassays. Quantitation was obtained by spectrometric analysis at OD 450 nm after addition of 200 microl of colorimetric substrate. Mean EGF levels ranged from 2441 pg/ml presurgically to 3349 pg/ml at 18 hours postsurgically and 1207 pg/ml at 6 weeks postsurgically. Repeated measures analysis of variance indicated statistically significant differences in 18 hours postsurgical salivary EGF levels when compared to controls and to postsurgical levels from 36 hours forward (P < 0.01). A second smaller rise in EGF was detected at 30 hours. These results suggest a transient increase in salivary EGF levels in response to intra-oral wounding.

Adult↗

PubMed1998

Direct synovial gene transfer with retroviral vectors in rat adjuvant arthritis.

OBJECTIVE: To evaluate the feasibility of direct in vivo gene transfer in an animal model of arthritis using a retroviral vector. METHODS: The timing and dose of retroviral vector was examined using very high titer retroviral vector (> or = 10(9) CFU) in rat adjuvant arthritis. Retroviral vector expressing beta-galactosidase (beta-gal) or vehicle alone was injected into the right ankle of rats with adjuvant arthritis. Ankles were injected either on Day 7 (pre-arthritis), Day 10 (early arthritis), Day 15 (accelerating arthritis), or Day 28 (chronic arthritis) after adjuvant immunization. Joints were harvested 3 days later and extracts were assayed for beta-gal activity. RESULTS: Synovial beta-gal expression was minimal in the Day 7 group and elevated in the Day 10, Day 15, and Day 28 groups. Gene transfer with retroviral vector did not exacerbate the local inflammatory response. Minimal or no beta-gal expression was observed in the contralateral uninjected paw or in the spleen, lung, liver, and kidneys. Frozen sections of retroviral vector injected joints were stained with X-gal and revealed transduced cells in the lining and superficial sublining layers. To determine the longevity of gene expression, ankle joints were injected with vector on Day 15 post-adjuvant, harvested, and assayed for beta-gal activity for up to 49 days after injection. Expression of the enzyme peaked from Day 3 to 7 and was still readily detected up to 49 days after retrovirus infection. CONCLUSION: This is the first report of successful direct in vivo gene transfer in the rat adjuvant arthritis model using a retroviral vector. Appropriate timing of administration and very high titer retroviral vector preparations are key determinants of adequate gene transduction.

Animals↗

PubMed1998

Excessive synthesis of matrix metalloproteinases in exocrine tissues of NOD mouse models for Sjögren's syndrome.

OBJECTIVE: Matrix metalloproteinases (MMP) and their substrates, components of the extracellular matrix, regulate environmental signals for cellular differentiation and tissue function. Changes in the levels of these enzymes may influence cell survival as well as pathology involving ectopic apoptosis. Using the non-obese diabetic (NOD) mouse model for Sjögren's syndrome, we evaluated the synthesis and expression of MMP in the exocrine target tissues of autoimmunity. METHODS: NOD, immunodeficient NOD-scid, and nondiabetic NOD.B10.H2b mice were evaluated for MMP activity in their saliva and exocrine gland lysates by gelatin zymography and reverse transcriptase-polymerase chain reaction (RT-PCR). In addition, changes in protein content of saliva and gland lysates were determined by specific Western blot and by enzymatic activity of amylase and cysteine proteases. Mice continuously treated with the MMP inhibitor GM6001 were evaluated from 7 to 20 weeks of age for the contribution of MMP activity to development of these hallmark biochemical markers of Sjogren's syndrome-like disease of NOD mice. RESULTS: Gelatin zymography of whole saliva and gland lysates indicated the presence of increased proteolytic activity, corresponding to proteins with a molecular mass ranging from 50 to 95 kDa, in the saliva of older (> 20 weeks of age) NOD mice as well as NOD.B10.H2b and NOD-scid mice compared to BALB/c controls. Elevated steady state levels of mRNA transcripts for the gelatinases MMP-2 and MMP-9 were detected in total RNA extracted from parotid and submandibular glands by RT-PCR. Despite prophylactic injection of the broad spectrum MMP inhibitor GM6001 into mice beginning at 7 weeks of age and continuing to 20 weeks, development of the autoimmune exocrinopathy was neither stopped nor retarded. CONCLUSION: These observations suggest that excessive MMP activity is associated with autoimmune Sjögren's syndrome-like disease in NOD mice. However, a possible contribution by increased MMP activity in initiation and progression of this autoimmune disease is yet to be elucidated.

Animals↗

PubMed1997

Prospective study of the natural history of thoracic aortic aneurysms.

BACKGROUND: The decision whether or not to recommend resection of moderately large descending thoracic and thoracoabdominal aneurysms requires weighing the relatively high mortality and significant risk of paraplegia associated with operation against the likelihood that the aneurysm will rupture spontaneously, with an almost invariably fatal outcome. To better define the risk of aneurysm rupture, we undertook a prospective study of patients who had not had operation on their moderately large descending thoracic and thoracoabdominal aneurysms. METHODS: Patients were enrolled at the time of their second computed tomographic scans: three-dimensional computer-generated reconstructions allowed determination of several dimensional parameters for each study, including diameters and cross-sectional areas at the site of maximal dilatation in the descending aorta and in the abdomen as well as total thoracoabdominal surface area. Comparisons of serial studies permitted calculation of yearly rates of change in these dimensions. RESULTS: Of 114 patients, 8 died of causes unrelated to the aneurysm, 26 died of rupture, 20 met previously determined criteria for operation, and 60 survived without operation or rupture. Multivariate regression analysis identified maximal diameter in the descending and in the abdominal aorta as independent risk factors for rupture, as well as older age, the presence of even uncharacteristic pain, and a history of chronic obstructive pulmonary disease. A piecewise exponential model enabled construction of an equation allowing calculation of rate of rupture in patients in whom the values of the risk factors are known, and also of the probability of rupture in a given individual over a specified time interval. CONCLUSIONS: Because using this equation--based on easily determined risk factors (age, pain, chronic obstructive pulmonary disease, maximal thoracic and maximal abdominal aortic diameter)--allows the risk of aneurysm rupture within a given interval to be estimated fairly accurately for each individual patient, it is our current practice to recommend operation when the calculated risk of rupture within 1 year exceeds the anticipated mortality of elective operation, rather than relying on general operative guidelines based almost exclusively on aneurysm size.

Adult↗

PubMed1997

PSP expression in murine lacrimal glands and function as a bacteria binding protein in exocrine secretions.

Nonobese diabetic (NOD) mice, an animal model for type I autoimmune diabetes and autoimmune sialoadenitis, abnormally express parotid secretory protein (PSP) in the submandibular glands (Robinson, C. P., H. Yamamoto, A. B. Peck, and M. G. Humphreys-Beher. Clin. Immunol. Immunopathol. 79: 50-59, 1996). To evaluate possible PSP gene dysregulation in the NOD mouse, we have examined a number of organs and tissues for PSP mRNA transcripts and protein expression. Results indicate that PSP is produced in the lacrimal glands of NOD mice as well as most laboratory mouse strains. Although purified salivary PSP from C3H/HeJ or BALB/c mice fails to affect amylase enzyme activity in in vitro assays, PSP bound to whole bacteria in a Zn2+-dependent manner. Additionally, radiolabeled protein bound to specific bacterial membrane proteins using a ligand binding assay. PSP gene transcription, but not protein production, was observed in the heart and pancreas from NOD mice, indicating abnormal transcription of the PSP gene. Sequence analysis of PSP cDNA from NOD mice revealed numerous base differences (compared with the published PSP sequence) capable of leading to significant amino acid substitutions, suggestive of strain-specific differences for the protein in mice. Together these results suggest that there exists in the NOD mouse a dysregulation of PSP transcription in various tissues. However, except for C3H/HeJ mice, PSP appears as a normal product of the lacrimal glands where, as in saliva, it may function as a nonimmune antimicrobial agent in the protection of tissue surfaces exposed to the external environment.

Adhesins, Bacterial↗

PubMed1997

Severe sore throat as a presenting symptom of adult onset Still's disease: a case series and review of the literature.

We describe our experience with 3 cases and a literature review examining whether sore throat is an early manifestation of adult onset Still's disease (AOSD). From our review of 341 cases in the English literature we noted that 69% of all reported patients displayed sore throat early in the disease course. We propose that sore throat be considered an important early diagnostic manifestation of AOSD because it appears useful in identifying patients in their first month of illness.

Adolescent↗

PubMed1996

Is vitrification involved in depression of the phase transition temperature in dry phospholipids?

Recent literature has suggested that the depression of the phase transition temperature (Tm) in dry phospholipids by sugars may be ascribed to vitrification of the stabilizing solute, rather than by the direct interaction between sugar and phospholipid we have proposed. Koster et al. ((1994) Biochim. Biophys. Acta 1193, 143-150) claim that the only necessity is that the glass transition (Tg) for the sugar exceed Tm for the lipid. Evidence is presented in the present paper that this is not sufficient. Based on the vitrification hypothesis of Koster et al., the predicted order of effectiveness in depressing Tm in dry dipalmitoylphosphatidylcholine (DPPC) is dextran > or = hydroxyethyl starch > stachyose > raffinose > trehalose > sucrose > glucose. In fact, the opposite order was seen. The effect of raffinose, sucrose, or trehalose on Tm in dry DPPC depends on the thermal history of the sample, as we have reported previously. When DPPC dried with trehalose is heated for the first time, Tm is about 55 degrees C, but on the second and subsequent heating scans Tm falls to about 25 degrees C. Koster et al. suggest that this effect is due to heating the sample above Tg rather than to melting the hydrocarbon chains. We present evidence here that all that is required is for the chains to be melted. Further, we show that retention of residual water by DPPC dried with trehalose depends on the drying temperature, but is independent of drying temperature with glucose, a finding that is consistent with direct interaction. We conclude that vitrification is not in itself sufficient to depress Tm in dry phospholipids.

1,2-Dipalmitoylphosphatidylcholine↗

PubMed1996

Alterations in the secretory response of non-obese diabetic (NOD) mice to muscarinic receptor stimulation.

Salivary gland secretion in non-obese diabetic (NOD) and BALB/c mice was evaluated following stimulation with the muscarinic receptor agonist pilocarpine. Both saliva flow rates and total protein were similar in BALB/c and prediabetic NOD mice. With diabetes onset in NOD mice, the saliva flow rate and protein concentration were dramatically reduced. The level of cyclic AMP (cAMP) generated 10 min following agonist injection was similar for the parotid gland of BALB/c and prediabetic and diabetic NOD mice but was reduced in diabetic NOD mice. In the submandibular gland both prediabetic and diabetic NOD mice showed a reduced potential for the generation of cAMP compared with BALB/c mice. The parotid gland from NOD mice had elevated levels of basal and muscarinic receptor agonist (carbachol)-stimulated concentrations of inositol phosphate intermediates in in vitro assays compared with BALB/c animals, while in the submandibular gland of NOD mice, basal and agonist-stimulated concentrations of inositol phosphate intermediates were reduced relative to BALB/c. An evaluation of muscarinic receptor density showed a reduction with diabetes onset for the parotid gland, while a similar level was observed in prediabetic NOD and BALB/c mice. The receptor density was decreased for both the prediabetic and diabetic NOD submandibular glands compared with BALB/c animals. Sera from diabetic NOD but not BALB/c mice immunoprecipitated radiolabeled muscarinic receptor, indicating the presence of autoantibody to the receptor in diabetic mice. Thus, the reduction of muscarinic agonist response in NOD mice may be due, in part, to a generalized reduction in signal transduction components most evident in the prediabetic and diabetic submandibular gland and to a less certain degree in the parotid gland from diabetic NOD mice as a consequence of autoantibodies directed against cell surface antigens.

Animals↗