Biomedical subjects

K Y Wu

Publications and source records attributed to K Y Wu.

At least 37 records · Page 2Linked to original sources

PubMed1998

Application of gas chromatography/electron capture negative chemical ionization high-resolution mass spectrometry for analysis of DNA and protein adducts.

The analytical potential of gas chromatography/electron capture negative chemical ionization high-resolution mass spectrometry (HRMS) for characterization and quantitation of DNA and hemoglobin adducts was demonstrated using three model compounds: N2, 3-ethenoguanine (EG), 7-(2-hydroxyethyl)guanine (7-HEG), and N-(2-hydroxyethyl)valine (HEV). At a resolving power of 10 000, the signal-to-noise (S/N) ratios obtained from quantitative selected ion monitoring (SIM) experiments using biological samples were comparable to or better than existing unit mass resolution experiments due to the reduction of chemical noise from the use of narrower mass windows. The specificity gained by HRMS was essential for quantitation of ultratrace amounts near the limit of detection since coeluting interferences of the analyte or internal standard can lead to inaccurate measurement of response factors. The limit of detection (LOD) was 100 amol (S/N = 5) using a pure standard of TTB2-EG. The LOD for complete assays using spiked samples was 500 amol (S/N = 5) for EG and 600 amol (S/N = 5) injected for 7-HEG. The standard deviation (SD) for the HRMS quantitative measurements was typically less than 10%. The SD for the complete biological assays as determined by spiking replicate samples was less than 15%. This method has adequate sensitivity and specificity to accurately measure DNA and protein adducts as low as endogenous concentrations in rodent and human tissues.

Animals↗

PubMed1998

An automated fluorescent PCR method for detection of shiga toxin-producing Escherichia coli in foods.

An automated fluorescence-based PCR system (a model AG-9600 AmpliSensor analyzer) was investigated to determine whether it could detect Shiga toxin-producing Escherichia coli (STEC). The AmpliSensor PCR assay involves amplification-mediated disruption of a fluorogenic DNA signal duplex (AmpliSensor) that is homologous to conserved target sequences in a 323-bp amplified fragment of Shiga toxin genes stx1, stx2, and stxe. Using the Amplisensor assay, we detected 113 strains of STEC belonging to 50 different serotypes, while 18 strains of non-Shiga-toxin-producing E. coli and 68 strains of other bacteria were not detected. The detection limits of the assay were less than 1 to 5 CFU per PCR mixture when pure cultures of five reference strains were used and 3 CFU per 25 g of food when spiked ground beef samples that were preenriched overnight were used. The performance of the assay was also evaluated by using 53 naturally contaminated meat samples and 48 raw milk samples. Thirty-two STEC-positive samples that were confirmed to be positive by the culture assay were found to be positive when the AmpliSensor assay was used. Nine samples that were found to be positive when the PCR assay was used were culture negative. The system described here is an automated PCR-based system that can be used for detection of all serotypes of STEC in food or clinical samples.

Animals↗

PubMed1998

Genomic analysis reveals chromosomal variation in natural populations of the uncultured psychrophilic archaeon Cenarchaeum symbiosum.

Molecular phylogenetic surveys have recently revealed an ecologically widespread crenarchaeal group that inhabits cold and temperate terrestrial and marine environments. To date these organisms have resisted isolation in pure culture, and so their phenotypic and genotypic characteristics remain largely unknown. To characterize these archaea, and to extend methodological approaches for characterizing uncultivated microorganisms, we initiated genomic analyses of the nonthermophilic crenarchaeote Cenarchaeum symbiosum found living in association with a marine sponge, Axinella mexicana. Complex DNA libraries derived from the host-symbiont population yielded several large clones containing the ribosomal operon from C. symbiosum. Unexpectedly, cloning and sequence analysis revealed the presence of two closely related variants that were consistently found in the majority of host individuals analyzed. Homologous regions from the two variants were sequenced and compared in detail. The variants exhibit >99.2% sequence identity in both small- and large-subunit rRNA genes and they contain homologous protein-encoding genes in identical order and orientation over a 28-kbp overlapping region. Our study not only indicates the potential for characterizing uncultivated prokaryotes by genome sequencing but also identifies the primary complication inherent in the approach: the widespread genomic microheterogeneity in naturally occurring prokaryotic populations.

Animals↗

PubMed1998

Ocular hypotensive and vasodilative effects of two beta-adrenergic blockers with intrinsic sympathomimetic activity.

PURPOSE: The ocular hypotensive and vasodilative effects of vaninolol and eugenolol, two beta-adrenergic blockers with intrinsic sympathomimetic activity, were tested in rabbits and their pharmacologic mechanisms were also studied in vitro. METHODS: Intraocular pressure was measured in ocular hypertonic rabbits which were induced by infusing 20% NaCl or 5% glucose solution. The rabbit's ocular blood flow was determined using the colored microsphere technique. The concentrations of cAMP were evaluated in porcine ciliary body and cultured A7r5 smooth muscle cells by radioimmunoassay. Ca+2 concentration was measured in A7r5 cells by spectrofluorometry after loading cells with Fura-2-AM. RESULTS: It was found that 0.5% eugenolol and vaninolol could suppress the intraocular pressure in glucose-induced ocular hypertensive rabbits and delay the intraocular pressure recovery in NaCl-induced ocular hypertensive rabbits. In addition, both agents improved the ocular blood flow in the iris, ciliary body, retina and choroid. Vaninolol and eugenolol of 10 microM inhibited the basal cAMP accumulation from 23.9 +/- 2.0 of control to 8.7 +/- 0.4 and 2.4 +/- 0.1 respectively and inhibited the isoproterenol-induced cAMP accumulation from 154.3 +/- 13.6 to 120.6 +/- 8.3 and 74.2 +/- 6.1 respectively in the porcine ciliary body. The cellular cAMP concentration was significantly increased from 10 +/- 1 of control to 96 +/- 5 (vaninolol) and 38 +/- 3 (eugenolol) in cultured A7r5 smooth muscle cells. Both agents also increased the intracellular calcium concentration in A7r5 cells. CONCLUSIONS: These results indicate that the lowering of intraocular pressure by vaninolol and eugenolol may be due to cAMP suppression in the ciliary body by beta-antagonist and/or alpha2-agonist activities. Both agents cause vasodilation via beta2-agonist action that increase the smooth muscle cellular cAMP level more than vasoconstriction via alpha-agonist activity by increasing an influx of extracellular Ca+2.

Adrenergic beta-Antagonists↗

PubMed1998

Calcium-induced changes on crystallins in organ-cultured porcine lens.

In this study, intact porcine lenses were cultured in vitro for 7 days supplemented with commercial balanced salt solution (BSS) which is usually used as an irrigation solution during intraocular surgery, and the lenses were maintained under various culture conditions, e.g. temperature and CO2 concentration. The intact porcine lenses after 7 days culture were analyzed with optical density scanner, gel permeation chromatography on TSK HM-55 column and SDS-PAGE (polyacrylamide gel electrophoresis). It was found that lenses exhibited the least opacity when lenses were cultured with Ca(+2)-free BSS buffer, CO2-free incubator and maintained at a temperature of 25 degrees C. After the lenses were cultured with Ca(+2)-free BSS or BSS medium, the composition of crystallins in lenses was separated with TSK HM-55 column. It was indicated that the percentage of high molecular weight (HMW) protein and (alpha-crystallin increased, and gamma-crystallin decreased in lenses incubated with BSS medium compared with lenses incubated with Ca(+2)-free BSS medium. Following an increase in the concentration of calcium in the medium from 4.3 mM, 20 mM, 50 mM, 100 mM to 200 mM, the opacity of the lens was measured with a densitometer. The changed percentage of various crystallins was similar to lenses with BSS media that increased in HMW protein and alpha-crystallin, decreasing in gamma-crystallin. In the case of lens protein pattern, the crystallin washed from TSK HM-55 gel was separated with SDS-PAGE (polyacrylamide gel electrophoresis). It was indicated that some of proteins disappeared when lenses were incubated with various concentrations of calcium. The vanished pH proteins were 20.5 kDa at 50 mM calcium, 20.5 kDa and 21 kDa at 100 mM, 20.5 kDa, 21 kDa, 22 kDa and 23 kDa at 200 mM which were compared with the protein bands in the presence of 20 mM calcium in BSS medium. This study indicates that the commercial balanced salt solution (BSS) which is usually used as an irrigating solution during intraocular operations may increase the risk for lens opacity because of the calcium contained in the solution.

Animals↗

PubMed1998

Dexon and nylon-sutured wound reaction in conjunctival flap after trabeculectomy combined with or without topical application of mitomycin-C.

In this study, rabbits were used to evaluate the sutured wound reaction with Dexon or nylon in the conjunctival flap 1, 4, 7, 14 and 28 days after trabeculectomy surgery with or without the use of mitomycin-C. Four major treated groups were used to compare their wound healing reaction; group 1--nylon-suture and non-mitomycin treatment; group 2--nylon-suture and mitomycin treatment; group 3--Dexon-suture and non-mitomycin treatment; group 4--Dexon-suture and mitomycin treatment. One day after surgery, the number of polymorphs was the greatest most in the nylon-sutured and non-mitomycin treated tissues (86 +/- 2). Four days after surgery, the number of polymorphs was the greatest most in Dexon-sutured and non-mitomycin treated tissues (109 +/- 87). The number of fibroblasts was the greatest most in nylon-sutured and non-mitomycin treated tissues (111 +/- 23). Seven days after surgery, the number of polymorphs was the greatest most in Dexon-sutured and mitomycin treated tissues (32 +/- 12). The number of fibroblasts was the greatest most in nylon-sutured and non-mitomycin treated tissues (126 +/- 15). Fourteen days after surgery, the number of fibroblasts was the greatest most in Dexon-sutured and non-mitomycin tissues (43 +/- 10). The number of goblet cells was the greatest most in nylon-sutured and non-mitomycin treated tissues (4 +/- 2). Twenty-eight days after surgery, the number of fibroblasts was the greatest most in Dexon-sutured and mitomycin treated tissues (40 +/- 15). The number of goblet cells was the greatest most in nylon-sutured and non-mitomycin treated tissues (4 +/- 2). Our conclusions are as follows: 1). The concentration of mitomycin in conjunctival wound edge should be maintained at as low a level as possible because the mitomycin will delay the wound healing process; 2). Nylon material is better than Dexon for conjunctival wound suture because nylon could induce a great quantity of fibroblasts before Dexon did.

Administration, Topical↗

PubMed1997

A rapid, sensitive and automated method for detection of Salmonella species in foods using AG-9600 AmpliSensor Analyzer.

The AG-9600 AmpliSensor Analyzer is an automated fluorescence-based system for detection of polymerase chain reaction (PCR) products. The principle of the AmpliSensor PCR assay involves amplification-mediated disruption of a fluorogenic DNA signal duplex (AmpliSensor) that is homologous to a target sequence within a 284-bp amplified fragment of the Salmonella invA gene. Since the assay is homogenous, the data can be obtained by direct measurement of fluorescence of the amplification mixture. The accumulation of the amplified product, reflected by the fluorescence index, is monitored cycle by cycle by the AG-9600 Analyzer. The detection limit of the assay was less than 2 colony-forming units (cfu) per PCR reaction using a pure culture of Salmonella typhimurium. In post-spiking experiments in which Salmonella was added to the overnight pre-enriched samples (chicken carcass rinses, ground beef, ground pork and raw milk), the detection limit of the assay was 2-6 cfu per PCR reaction. In pre-spiking experiments in which Salmonella was added to the samples prior to overnight pre-enrichment, the detection limit was less than 3 cfu per 25 g or 25 ml of food. The assay was up to 2 orders of magnitude more sensitive than detection by ethidium bromide-stained agarose gel electrophoresis. To further evaluate assay performance, 54 naturally contaminated chicken carcass rinses, 65 raw milk and six ground pork samples were tested in the study. Thirty-eight Salmonella-positive samples confirmed by the Modified Semi-solid Rappaport-Vassiliadis (MSRV) culture assay were found positive using the AmpliSensor assay. Two chicken carcass rinses found positive using the assay were MSRV-negative. In addition, relative quantification using the AmpliSensor assay was linear up to 3 logs of initial target concentration in artificially contaminated food samples.

Animals↗

PubMed1997

Effects of pilocarpine and other antiglaucoma drugs on cultured human Tenon's fibroblast cells.

It has been found that long-term therapy with topical antiglaucoma drugs may decrease the success of glaucoma filtering surgery. In this study, various antiglaucoma drugs, including carteolol, betaxolol, levobunolol, pilocarpine, and timolol, were investigated for their proliferative effect on cultured human Tenon's fibroblast cells. It was found that the 3H-thymidine uptake of cultured fibroblast cells was inhibited by commercial antiglaucoma drugs, including carteolol of 0.1% concentration (13%), 0.01% (50%) and 0.001% (53%), betaxolol of 0.05% concentration (14%), 0.005% (42%) and 0.0005% (62%), levobunolol of 0.05% concentration (2%), 0.005% (32%) and 0.0005% (55%), and timolol of 0.025% concentration (4%), 0.0025% (47%) and 0.00025% (55%), whereas the 3H-thymidine uptake was increased by commercial pilocarpine eyedrop from 103% (0.2% of concentration), 170% (0.02% of concentration) to 171% (0.002% of concentration), when cells were treated with commercial drugs for 100 min. Meanwhile, the proliferations of cultured fibroblast cells were stimulated by simultaneously combining 0.2%, 0.02% and 0.002% concentrations of pilocarpine eyedrops with other antiglaucoma drugs, such as carteolol of 0.1% concentration (50%), 0.01% (113%) and 0.001% (138%), betaxolol of 0.05% concentration (24%), 0.005% (128%) and 0.0005% (142%), levobunolol of 0.05% concentration (32%), 0.005% (87%) and 0.0005% (119%), and timolol of 0.025% concentration (15%), 0.0025% (94%) and 0.00025% (118%). Following incubation with pure pilocarpine, the proliferation of Tenon's fibroblast cells was inhibited for 84% (0.2% concentration), 84% (0.02% concentration) and 90% (0.002% concentration). When fibroblast cells were treated with commercial pilocarpine eyedrops for 24 hours, the 3H-thymidine uptake was increased to 160% (0.02% concentration) and 172% (0.002% concentration). This result shows that the commercial pilocarpine may play a crucial role for the proliferation of cultured human Tenon's fibroblast cells.

Adrenergic beta-Antagonists↗

PubMed1997

Effects of antiglaucoma drugs and corticosteroids on the cellular glucose uptake in cultured porcine corneal endothelial cells.

In this study, various commercially used antiglaucoma drugs and corticosteroids were investigated in their effects on porcine corneal endothelial cells especially in cellular glucose uptake. Cellular glucose uptake directly affects the pumping efficiency in corneal endothelial cells. Following the cells' treatment with various antiglaucoma eyedrops for 100 min, the 3H-2-deoxyglucose uptake in cultured porcine corneal endothelial cells was affected by betaxolol from 3.1% (1.6 mM), 181% (0.16 mM) to 158% (0.016 mM), by timolol from 93% (0.79 mM), 227% (0.079 mM) to 151% (0.0079 mM), by carteolol from 141% (3.4 mM), 180% (0.34 mM) to 97% (0.034 mM), by levobunolol from 80% (1.5 mM), 98% (0.15 mM) to 90% (0.015 mM), by dipivefrin from 116% (0.2 mM), 176% (0.02 mM) to 108% (0.002 mM) and by pilocarpine from 115% (9.6 mM), 210% (0.96 mM) to 210% (0.096 mM) when the cells were compared with a control medium. In the presence of various corticosteroids, the glucose uptake in corneal endothelial cells was affected by fluorometholone from 160% (0.26 mM), 139% (0.026 mM) to 107% (0.0026 mM), by dexamethasone from 85% (0.25 mM), 117% (0.025 mM) to 109% (0.0025 mM) and by betamethasone from 95% (0.25 mM), 96% (0.025 mM) to 99% (0.0025 mM). These results show that the commercial eyedrops of antiglaucoma drugs and corticosteroids will not decrease the cellular glucose uptake in cultured porcine corneal endothelial cells except when incubated with high concentrations of betaxolol, levobunolol and dexamethasone.

Adrenal Cortex Hormones↗

PubMed1997

Alteration of glucose uptake in cultured human corneal endothelial cells grown in high glucose media via cAMP-dependent pathway.

In this study, cultured human corneal endothelial cells were incubated in media containing various concentrations of glucose at 5 mM, 10 mM, and 25 mM for 2 days. Then, the cellular 2-deoxyglucose uptake and cAMP concentration of cultured human corneal endothelial cells were measured. The results indicated that the activity of cellular glucose uptake of nmole/min/mg protein was decreased gradually from 0.18 (5 mM), 0.10 (10 mM), 0.07 (20 mM) to 0.06 (25 mM) after 2 days incubation with a high concentration of glucose. The glucose uptake in insulin-treated human corneal endothelial cells also exhibited a similar declining effect in high glucose media from 0.30 (5 mM), 0.11 (10 mM), 0.08 (20 mM) to 0.05 (25 mM). The cAMP concentration in human corneal endothelial cells was measured in the presence of high glucose media. It was indicated that the cAMP concentrations of pmole/well in both insulin-treated and non-insulin treated cells were also decreased after increasing the glucose concentration in the media from 73 (5 mM) to 20 (25 mM) and 101 (5 mM) respectively. The cAMP concentration in insulin-treated cells was less than in non-insulin treated cells. This decreasing effect was significantly reversed by the addition of 1 mM dibutyryl-cAMP to the cells for 1 hour in both groups. These results suggest that the diabetic state may decrease the 2-deoxyglucose uptake in human corneal endothelial cells via cAMP-dependent pathway.

Animals↗

PubMed1996

A psychrophilic crenarchaeon inhabits a marine sponge: Cenarchaeum symbiosum gen. nov., sp. nov.

Archaea, one of the three major domains of extant life, was thought to comprise predominantly microorganisms that inhabit extreme environments, inhospitable to most Eucarya and Bacteria. However, molecular phylogenetic surveys of native microbial assemblages are beginning to indicate that the evolutionary and physiological diversity of Archaea is far greater than previously supposed. We report here the discovery and preliminary characterization of a marine archaeon that inhabits the tissues of a temperate water sponge. The association was specific, with a single crenarchaeal phylotype inhabiting a single sponge host species. To our knowledge, this partnership represents the first described symbiosis involving Crenarchaeota. The symbiotic archaeon grows well at temperatures of 10 degrees C, over 60 degrees C below the growth temperature optimum of any cultivated species of Crenarchaeota. Archaea have been generally characterized as microorganisms that inhabit relatively circumscribed niches, largely high-temperature anaerobic environments. In contrast, data from molecular phylogenetic surveys, including this report, suggest that some crenarchaeotes have diversified considerably and are found in a wide variety of lifestyles and habitats. We present here the identification and initial description of Cenarchaeum symbiosum gen. nov., sp. nov., a symbiotic archaeon closely related to other nonthermophilic crenarchaeotes that inhabit diverse marine and terrestrial environments.

Animals↗

PubMed1996

Use of a fluorescent-PCR reaction to detect genomic sequence copy number and transcriptional abundance.

We present a fluorescent-PCR-based technique to assay genomic sequence copy number and transcriptional abundance. This technique relies on the ability to follow fluorescent PCR progressively in real time during the exponential phase of the reaction so that quantitative PCR is accomplished. We demonstrated the ability of this technique to quantitate both known deletions and amplifications of loci that have been measured previously by other methods, and to measure transcriptional abundance. Using an efficient variant of the fluorescent-PCR technology, we can monitor transcription semiquantitatively. The ability to detect all amplifications and deletions at any single copy locus by PCR makes this the technique of choice to assay genomic sequence copy number anomalies in birth defects and cancers. The ability to detect variations in transcript abundance enables this technique to fashion a time and tissue analysis of transcription.

Chromosome Aberrations↗

PubMed1996

Characterization of uncultivated prokaryotes: isolation and analysis of a 40-kilobase-pair genome fragment from a planktonic marine archaeon.

One potential approach for characterizing uncultivated prokaryotes from natural assemblages involves genomic analysis of DNA fragments retrieved directly from naturally occurring microbial biomass. In this study, we sought to isolate large genomic fragments from a widely distributed and relatively abundant but as yet uncultivated group of prokaryotes, the planktonic marine Archaea. A fosmid DNA library was prepared from a marine picoplankton assemblage collected at a depth of 200 m in the eastern North Pacific. We identified a 38.5-kbp recombinant fosmid clone which contained an archaeal small subunit ribosomal DNA gene. Phylogenetic analyses of the small subunit rRNA sequence demonstrated it close relationship to that of previously described planktonic archaea, which form a coherent group rooted deeply within the Crenarchaeota branch of the domain Archaea. Random shotgun sequencing of subcloned fragments of the archaeal fosmid clone revealed several genes which bore highest similarity to archaeal homologs, including large subunit ribosomal DNA and translation elongation factor 2 (EF2). Analyses of the inferred amino acid sequence of archaeoplankton EF2 supported its affiliation with the Crenarchaeote subdivision of Archaea. Two gene fragments encoding proteins not previously found in Archaea were also identified: RNA helicase, responsible for the ATP-dependent alteration of RNA secondary structure, and glutamate semialdehyde aminotransferase, an enzyme involved in initial steps of heme biosynthesis. In total, our results indicate that genomic analysis of large DNA fragments retrieved from mixed microbial assemblages can provide useful perspective on the physiological potential of abundant but as yet uncultivated prokaryotes.

Amino Acid Sequence↗

PubMed1996

Effects of dopamine antagonists and agonist on cultured human Tenon's fibroblast cells.

It was found that long-term therapy with topical antiglaucoma drugs may increase the number of fibroblasts and inflammatory cells in the conjunctiva and Tenon's capsule then decrease the success of glaucoma filtering surgery. Since some dopamine antagonists and agonists are reported to have ocular hypotensive effects, they could potentially be used as clinical antiglaucoma agents in future. In this study, several dopamine antagonists and agonists were evaluated in their effects on the fibroblast proliferation of ocular tissues with cultured human ocular Tenon's fibroblast cells. It was found that dopamine antagonists inhibited [3H]-thymidine uptake to 10.8%, 14.4%, 42.2% and 72.4% of control in domperidone, 28.6%, 67.7%, 96.9% and 99.2% in haloperidol, 9.8%, 18.9%, 103.5% and 104.6% in moperone and 9.8%, 24.7%, 70.8% and 96.9% in loxapine respectively, while the 0.5% concentration of drugs were dissolved in dimethyl sulfoxide (DMSO) followed by dilution in culture medium to final concentrations ranging from 0.05%, 0.005%, 0.0005% to 0.00005%. In the case of dopamine agonist, bromocriptine, the proliferation of cultured human Tenon's fibroblast cells was also inhibited for 25.1% of the control at 0.05% concentration and 92.3% of the control at 0.00005% concentration. These results indicate that these dopamine drugs have a potential long-term application in the treatment of glaucoma without having the stimulating effect on the proliferation of human Tenon's fibroblast cells.

Cells, Cultured↗

PubMed1995

Physical analysis of the tuberous sclerosis region in 9q34.

We report the construction of a physical map based on cloned DNA within the candidate region for the tuberous sclerosis complex (TSC1) gene on chromosome 9q34, between the markers D9S149 and D9S66. The DNA clones form three contigs consisting of 7 YACs, bridged by P1 and cosmid clones, and cover more than 950 kb of 9q34. Despite intensive screening of all available libraries, two gaps remain. A detailed physical map of much of this region was derived, and restriction mapping of the YAC, P1, and cosmid clones reveals novel CpG islands in this region. This set of genomic clones provides a resource for characterizing candidates for the TSC1 gene, guided by the location of CpG islands.

Base Sequence↗

PubMed1995

Dose-response relationships for carcinogens.

Biotransformation of chemical carcinogens involves both metabolic activation and detoxication. The molecular dose present on DNA as adducts represents a balance between these two pathways (formation) and DNA repair. All of these are enzymatic processes subject to saturation. When none of the pathways is saturated, linear molecular dosimetry is expected, whereas if metabolic activation is saturated, a supralinear response occurs. If detoxication or DNA repair is saturated, a sublinear response occurs. With chronic exposure, steady-state concentrations of DNA adducts develop and these follow the same patterns. With several alkylating agents, multiple adducts are formed. The extent of formation is chemically defined, but different DNA repair pathways can be involved for different adducts. By understanding the molecular dose and biology of each adduct and comparing these to the dose-response for tumor induction, it may be possible to identify the most appropriate biomarkers for risk assessment. Recently, endogenous DNA adducts identical to those induced by known human carcinogens have been identified. These endogenously formed adducts may play an important role in human carcinogenesis.

Animals↗

PubMed1994

High abundance of Archaea in Antarctic marine picoplankton.

Archaea (archaebacteria) constitute one of the three major evolutionary lineages of life on Earth. Previously these prokaryotes were thought to predominate in only a few unusual and disparate niches, characterized by hypersaline, extremely hot, or strictly anoxic conditions. Recently, novel (uncultivated) phylotypes of Archaea have been detected in coastal and subsurface marine waters, but their abundance, distribution, physiology and ecology remain largely undescribed. Here we report exceptionally high archaeal abundance in frigid marine surface waters of Antarctica. Pelagic Archaea constituted up to 34% of the prokaryotic biomass in coastal Antarctic surface waters, and they were also abundant in a variety of other cold, pelagic marine environments. Because they can make up a significant fraction of picoplankton biomass in the vast habitats encompassed by cold and deep marine waters, these pelagic Archaea represent an unexpectedly abundant component of the Earth's biota.

Animals↗

PubMed1994

Activity of growth hormone peptides bGH 96-133 and hGH 95-133 in 3T3-F442A cells.

Chemically synthesized bovine growth hormone (bGH) bGH 96-133 and its human homologue, hGH 95-133, have similar in vitro biological activities. Unlike native GH, bGH 96-133 and hGH 95-133 were completely without adipogenic or anti-insulin activity at doses up to 10 microM. bGH 96-133 had insulin-like activity, with a 100% increase in glucose uptake at 10 microM. bGH was anti-mitogenic and bGH 96-133 and hGH 95-133 were mitogenic (EC50 approximately 180 nM and maximal response at 1-2 microM). Only bGH 96-133 and hGH 95-133 displaced [125I]hGH 95-133 binding from 3T3-F442A fibroblasts with a Kd between 60-120 nM. bGH, hGH, insulin and IGF-I were without effect on [125I]hGH 95-133 binding. bGH 96-133 and hGH 95-133 did not significantly inhibit [125I]hGH or [125I]IGF-I binding. These experiments indicate that GH containing peptides bGH 96-133 and hGH 95-133 have mitogenic and insulin-like activity without the adipogenic, anti-insulin or anti-mitogenic activity of bGH. These peptides have a specific binding site which appears to be distinct from the GH, insulin and IGF-I receptors.

3T3 Cells↗