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Biomedical subjects

D Shen

Publications and source records attributed to D Shen.

At least 19 recordsLinked to original sources

Identification of the polypeptides of the major light-harvesting complex of photosystem II (LHCII) with their genes in tomato.

Using an improved SDS-PAGE system, the polypeptides of the major chlorophyll a/b light-harvesting complex of PSII (LHCII) from tomato leaves were resolved into five polypeptide bands. All the polypeptides were matched with the genes encoding them by comparing amino acid sequences of tryptic peptides with gene sequences. The two major LHCII bands (usually comigrating as a '27 kDa' polypeptide) were encoded by cab1 and cab3 (Type I LHCII) genes. A third strong band of about 25 kDa was encoded by cab4 (Type II) genes. Polypeptides from two minor bands of 23-24 kDa were not N-terminally blocked; their N-terminal sequences showed they were Type III LHCII proteins. One complete cDNA clone and several incomplete clones for Type III polypeptides were sequenced. Combined with the peptide sequences, the results indicate that there are at least four different Type III genes in tomato, encoding four almost identical polypeptides. Thus, all the LHCII CAB polypeptides have been identified, and each type of LHCII polypeptide is encoded by distinct gene or genes in tomato.

Amino Acid Sequence

Nucleotide sequence and chromosomal location of Cab11 and Cab12, the genes for the fourth polypeptide of the photosystem I light-harvesting antenna (LHCI).

Tryptic peptide sequences from the 22 kDa polypeptide of tomato LHCI were used to construct a probe for gene cloning. The two genes cloned, cab11 and cab12, encode proteins of 251 and 250 residues that are 88% identical in overall amino acid sequence and 93% identical in the deduced mature protein. Each gene is present in a single copy per haploid genome; cab11 on chromosome 3 and cab12 on chromosome 6, and each has 2 introns located in similar positions to introns in other members of the Chl a/b-binding (CAB) protein gene family. Comparison of the amino acid sequences of LHCI, LHCII, CP29 and CP24 polypeptides confirms that all CABs share two regions of very high similarity which include the first and third transmembrane helices and the stroma-exposed sequences preceding them. However, near the N-terminus and between the conserved regions, the LHCI polypeptides have sequence motifs which appear to be PSI-specific.

Amino Acid Sequence

Proteolytic processing in a non-lysosomal compartment is required for transcytosis of protein-polylysine conjugates in cultured Madin-Darby canine kidney cells.

The transcytosis of horseradish peroxidase, as well as its poly(L-lys) and poly(D-lys) thioether conjugates, was investigated in Strain I Madin-Darby canine kidney (MDCK) cell monolayers grown on 0.4 microns pore size polycarbonate membranes in Costar Transwells. The 3 types of HRP had almost identical rates of transport during the first 2 hr of incubation. However, a significant increase of basal-to-apical transport was detected beginning at 3 hr only in Transwells containing the poly(L-lys) conjugate. This increase was inhibited by colchicine (2 microM) and by the Bowman-Birk protease inhibitor (0.1 mg/ml), but not by NH4Cl (10 mM) or chloroquine (0.1 mM). The increase was abolished either by prior trypsinization of the conjugate or by incubation at 4 degrees C. Ultrafiltration studies indicated that the transcytosed poly(L-lys) conjugate was smaller in size than the original conjugate. These results indicate that the conjugate was processed during transcytosis in a non-lysosomal proteolytic compartment, where its poly(L-lys) moiety was selectively degraded, allowing active peroxidase to be released into the apical medium.

Animals

In vitro synthesis of nitroxide free radicals by hog liver microsomes.

The in vitro biooxidation of 4-hydroxy-2,2,6,6-tetramethylpiperidine (TEMP), 4-hydroxy-2,2,4,4-tetramethyl-1,3-oxazolidine (TEMO) and diphenylamine (DPA) by hog liver microsomes to their respective nitroxide free radicals, 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO), 2,2,4,4-tetramethyl-1,3-oxazolidine-1-oxyl (TEMOO), and diphenylnitroxide (DPNO) has been investigated. For extending the life span of the liver microsomes, a calcium alginate immobilization procedure was used. The biooxidation rates of the above amines to their respective nitroxide metabolites were measured by means of oxygen uptake at 37 degrees C and pH 7.4. N-octylamine was found to be an activator in the biooxidation of the amines. The formation of the nitroxide radicals was identified by E.S.R. spectroscopy.

Alginates

Applications of a new chemiluminometric thyrotropin assay to subnormal measurement.

A new immunochemiluminometric TSH assay (ICMA) was shown to offer improved analytical (+2 SD of zero) and functional (20% interassay coefficient of variation) sensitivity [0.003 vs 0.045 +/- 0.005 (+/- SE; range, 0.01-0.07); 0.018 vs. 0.23 +/- 0.02 (range, 0.10-0.35, mU/L); analytical vs. functional sensitivity limit for the ICMA vs. 10 other TSH immunometric assays, respectively]. The ICMA was used to study the physiological relationship between serum TSH and free T4 [as reflected by free T4 index (FT4I)] values at both steady state and 14 days after acute pharmacological T4 administration (3 mg oral T4 load plus 0.3 mg daily). At steady state, an inverse log/linear relationship was found between serum TSH and FT4I values (log TSH = 2.56 - 0.022 FT4I; r = 0.84; P less than 0.001). Ten to 14 days after acute T4 suppression in 5 euthyroid subjects, serum TSH/FT4I levels had plateaued after decreasing in parallel to the slope of the steady state relationship, suggesting that the degree of T4 suppression of TSH can be predicted from an individual's pituitary TSH/free T4 set-point and the magnitude of the serum T4 elevation achieved. Ambulatory and hospitalized patient sera, previously identified as having low (less than 0.1 mU/L) TSH levels by a less sensitive assay, were restudied by the TSH ICMA. Normal TSH values ranged from 0.39-4.6 mU/L, whereas the majority of hyperthyroid patients [52 of 54 (96% ambulatory) and 22 of 23 (96%, hospitalized)] had undetectable (less than 0.005 mU/L), basal TSH levels and absent TRH stimulated TSH responses. In contrast, most (32 of 37; 86%) of hospitalized nonhyperthyroid patients with low (less than 0.1 mU/L) TSH values due to nonthyroidal illness or glucocorticoid treatment had detectable (greater than 0.01 mU/L) basal and TRH stimulated TSH levels. The positive relationship between basal and TRH-stimulated TSH levels was shown to extend down to the detectability limit of the assay (0.005 mU/L), which further supported the authenticity of the subnormal TSH ICMA measurements. The new TSH ICMA is considered to represent the first of a third generation of clinical TSH assays, since it has a functional (interassay) sensitivity that is 2 orders of magnitude greater than that of typical first generation TSH RIAs and 1 order of magnitude greater than current second generation TSH immunometric methods. Such third generation TSH assays will facilitate both the optimization of T4 therapy as well as the diagnosis of hyperthyroidism in hospitalized patients with nonthyroidal illness.

Adult

[Changes in the levels of human platelet membrane glycoproteins in patients with a new-type defect of platelet aggregation].

Labeled avidin-biotin (LAB)-enzyme linked immunosorbent assay (ELISA) for quantitative measurement of the levels of human platelet membrane glycoproteins (GPs) Ib, IIb and IIIa was developed by using a group of murine monoclonal antibodies raised against human platelet membrane GPs. The contents of the GPs Ib, IIb and IIIa in patients with a new-type defect of platelet aggregation were detected and found to be in normal range. However, the amounts of GPs IIb and IIIa before washing in patients were significantly decreased in these patients when compared with those in controls (P less than 0.01), while after washing, normal levels could be obtained. The results reported have support the hypothesis that the pathogenesis of this defect in platelet aggregation seems not to be associated with the abnormality of the levels of GPs, but is a result of abnormal factor existing in the plasma. Further investigation on the properties of the factor is in progress.

Blood Platelet Disorders

[An observation of protective effect of acupuncture on the gastric mucosa of Wistar rats and the relative histochemical changes of the neurotransmitters].

In this paper, changes of neurotransmitters and the protective effect of acupuncture on gastric mucosa have been studied by the histochemical methods of cholinesterase and catecholamine on 30 pairs of Wistar rats. 1. Sequential medical trials on rats of water-immersion induced gastric ulceration show that there was a protective effect of electro-acupuncture on gastric mucosa and it was statistically significant. 2. Under the protective effect of acupuncture on gastric mucosa, finally, both the cholinergic and adrenergic nerves undergo significant inhibition.

Acupuncture Therapy

[Binding of acid mucopolysaccharides of Stichopus japanicus to washed human platelets].

The binding of acid mucopolysaccharides of Stichopus japonicus Selenka (Sjamp) to washed human platelets was studied using [3H]Sjamp. The binding was rapid, reversible and with a saturable tendency. The binding reached to equilibrium at about 10 min, and 50% of [3H]Sjamp bound to the platelets at equilibrium within the first minute. When excess unlabeled Sjamp was added to the [3H]Sjamp binding suspension, [3H]Sjamp dissociated from the platelets rapidly and rather completely. Scatchard analysis revealed a class of binding sites with Kd = 2.3 +/- 0.7 micrograms/ml and Bmax = 4.5 +/- 1.2 micrograms/10(9) platelets. Separate experiments showed that the monoclonal antibody (McAb) SZ-21, a McAb to platelet membrane glycoprotein (GP) IIIa, weakened the binding, while McAb SZ-2 and SZ-22 (to GP Ib and GP IIb respectively) were ineffective. We also observed that heparin inhibited [3H]Sjamp binding to the platelets. These results indicated that the binding sites on platelets might be the Sjamp receptors. We conclude that receptor-Sjamp binding is an important process in platelet aggregation induced by Sjamp.

Animals

[Saliva phenobarbital concentration in epileptics].

The concentrations of phenobarbital were measured in saliva-serum pairs obtained simultaneously from 33 epileptics by RIA. The mean concentration of phenobarbital in saliva was 3.2 +/- 1.78 micrograms/ml, in serum was 11.75 +/- 7.4 micrograms/ml, the mean saliva-to-serum ratio for phenobarbital was 0.27 +/- 0.08. The results suggested that saliva concentration of phenobarbital was found to be correlated closely with the total serum level (R = 0.7922, P less than 0.01). The high correlation between saliva and serum concentrations supports the use of saliva measurement in epileptics. The simplicity and convenience of saliva collection and its noninvasiveness make it represent serum concentration of phenobarbital as an efficient monitoring method.

Adolescent

Nasal pharmacodynamics of brompheniramine in perennial rhinitis.

This study evaluated the relationship between brompheniramine maleate and changes in nasal reactivity. Ten subjects with moderate-to-severe perennial rhinitis took brompheniramine for seven days using either a standard formulation or sustained-release preparation. Nasal aerodynamics and response to histamine were assessed at the end of the week. Despite varying doses of brompheniramine maleate (12 to 32 mg/d), there was no significant difference in nasal reactivity to histamine or in changes of nasal airflow, indicating that low doses of brompheniramine are highly effective in blocking histamine, receptors in the nasal mucosa.

Adolescent

Levobunolol compared with timolol: a four-year study.

Fifty-one patients with raised intraocular pressure (IOP) were treated for up to four years with one of three ophthalmic solutions: 0.5% levobunolol, 1% levobunolol, or 0.5% timolol. The study was conducted as a double-masked, randomised trial in which medications were administered twice daily to both eyes. Levobunolol and timolol were equally effective in reducing overall mean IOP; reductions were greater than 8.8 mmHg in all three treatment groups. The study showed levobunolol to be as safe and effective as timolol in the long-term control of raised IOP.

Aged

Specificity of sensitive assays of thyrotropin (TSH) used to screen for thyroid disease in hospitalized patients.

Thyrotropin (TSH) concentrations were measured in 1580 hospitalized patients and 109 normal persons. Using the mean +/- 3 SD limits of the log values for the controls (0.35-6.7 milli-int. units/L), the proportion of abnormal TSH results in the hospitalized patients was 17.2%. TSH was undetectable (less than 0.1 milli-int. unit/L) in 3.1% of patients, suggesting hyperthyroidism, and high (greater than 20 milli-int. units/L) in 1.6%, suggesting hypothyroidism. On follow-up of 329 patients, 62% with abnormal TSH (less than 0.35 or greater than 6.7 milli-int. units/L) and 38% with normal TSH concentrations, only 24% of those with undetectable TSH had thyroid disease: 36% of them were being treated with glucocorticoids and 40% had nonthyroidal illness (NTI). Although half the patients with TSH greater than 20 milli-int. units/L had thyroid disease, 45% of patients had high TSH values associated with NTI. TSH concentrations usually returned towards normal when patients' therapy with glucocorticoids was discontinued or they recovered from NTI. TSH test sensitivity appeared good when the mean +/- 3 SD limits of the reference population were used, i.e., no cases of hyper- or hypothyroidism, as identified by free thyroxin index (FT4I), were missed. However, TSH test specificity was inferior to that of the FT4I test (90.7% vs 92.3%), although specificity could be improved to 97.0% if the wider TSH reference limits of 0.1 to 20 milli-int. units/L were used--limits considered pathological if applied to outpatients. Evidently, different reference intervals for TSH are needed for hospitalized and nonhospitalized patients. We conclude that a "sensitive TSH assay" is not a cost-effective thyroid screening test for hospitalized patients as compared with the FT4I.

Adult

The role of transferrin in natural killer cell and IL-2-induced cytotoxic cell function.

The growth factor transferrin (Tf) enhanced natural killer (NK) cell cytotoxicity. This enhancement was due to direct effects on NK cell function, and Tf treatment of the K562 target cell had no effect on their sensitivity. NK cells were highly enriched in the low-density large granular lymphocyte population (LGL) by Percoll gradient centrifugation. Despite the direct effect of Tf on NK cells, the number of cells expressing receptors for Tf (TfR) in NK-enriched LGL was the same as the NK-cell-depleted high-density small lymphocyte population (SL). All populations, tested without stimulation, had very few TfR+ cells. Interleukin 2 (IL-2) could induce very high NK-like activity in the LGL but not in SL. Similarly, only LGL could be induced by IL-2 to express TfR. In serum-free cultures, only limited NK-like activity could be developed which was greatly enhanced by supplementing with Tf in the cultures. The importance of Tf in NK-like development was confirmed by modulating the expression of TfR in IL-2 containing cultures with mouse monoclonal antibody OKT9 specific for TfR. OKT9 totally abrogated the induction of cytotoxic activity by IL-2 against K562 and NK-resistant target. OKT9 inhibited the induction of cytotoxicity in both lymphocytes containing active NK cells and in those predepleted of active NK cells, indicating that the development of NK-like activity from both precursor populations requires Tf. The inhibition by OKT9 was only during the induction phase. The same antibody had no effect on the cytotoxicity of fresh NK cells or the mature IL-2-induced NK-like cells. Our data therefore do not support the hypothesis of TfR as the NK recognition structure. Instead, these results indicate that Tf is important for the development of NK and NK-like activities.

Antibodies, Monoclonal