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Biomedical subjects

C Hunt

Publications and source records attributed to C Hunt.

At least 73 records · Page 4Linked to original sources

The translocation of mitochondria along insect ovarian microtubules from isolated nutritive tubes: a simple reactivated model.

Nutritive tubes, the microtubule-based translocation channels that link the trophic tissue to the developing oocytes in the ovaries of hemipteran insects, have been isolated and examined using video-enhanced differential interference contrast microscopy. When viewed in this way the nutritive tubes are seen to fray into linear strands, which, on the addition of exogenous ATP, support the translocation of particles along their lengths. The movement is also seen with GTP but not AMP-PNP. It is not affected by the addition of inhibitors of dynein or of energy metabolism. Electron microscopy shows the strands to consist of bundles of parallel microtubules of different sizes and the moving particles to be mitochondria. Comparisons are drawn between the movement of mitochondria along isolated insect ovarian microtubules and the reported translocation of vesicles along microtubules from squid axoplasm. The simplicity of the insect system is emphasized. The fact that it can be isolated easily and characterized biochemically makes it potentially valuable for investigating microtubule-based translocation.

Animals↗

Alcohol intake in a healthy elderly population.

The relationship between amount of alcohol consumption and social, psychological, and cognitive status was examined in 270 healthy, independently living men and women over age 65. Forty-eight per cent of the sample recorded some alcohol intake during a three-day diet record, with 8 per cent drinking 30 or more grams of alcohol daily. Alcohol intake was positively associated with male gender, income, and amount of education and negatively associated with age. Alcohol intake was not associated with any changes in social or psychological status, but was positively associated with several measurements of cognitive status. These correlations were weak, however, and tended to disappear after controlling for income, education, gender, and age. Past alcohol intake was not associated with any indicators of present social, psychological, or cognitive functioning.

Age Factors↗

[Comparison of the increase of stiffness produced during sinusoidal stretch of the long fibula muscle in the cat by the contraction of slow or fast motor units].

The increase in stiffness of cat peroneus longus muscle elicited by the contraction of homogeneous groups of motor units (slow, fast fatigue-resistant and fast fatigable) was measured during sinusoidal stretches (20-80 Hz) of small amplitude (40-100 micron). For comparable tetanic tensions, slow units increase muscle stiffness more than fast unit.

Animals↗

Organization, nucleotide sequence, and transcription of the chicken HSP70 gene.

We have isolated a gene encoding a 70,000-Da heat shock protein (HSP70) from a chicken genomic library. The recombinant clone C411 was isolated by hybridization to a radioactively labeled plasmid containing Drosophila HSP70 gene sequences. The identity was established by hybrid selected translation using a subcloned restriction fragment, pC1.8, and total cytoplasmic RNA from chicken cells. The selected mRNA translated in vitro a product which co-migrates with in vivo synthesized chicken HSP70 by SDS-polyacrylamide gel electrophoresis. By Northern blot analysis of poly(A)+ mRNA from heat-shocked cells, we detected a 2.6-kilobase pair mRNA homologous to pC1.8. The nucleotide sequences within pC1.8 and a flanking 2.0-kilobase pair HindIII fragment contain a single contiguous open reading frame of 1905 nucleotides that corresponds to a protein of predicted size 70,000 daltons. Upstream of the initiator methionine codon, ATG, lie sequences homologous to the canonical transcription elements TATA, CCAAT, and SP1 binding site, and two overlapping heat shock elements. The order and spacing of these sequences share many features in common with the promoter for the human HSP70 gene.

Amino Acid Sequence↗

Stage at diagnosis of cancer varies with the age of the patient.

The stage of a cancer at diagnosis is multiply determined, theoretically depending on such disparate factors as tumor biology, patient education, and physician behaviors. Data from all Hispanic and non-Hispanic white residents of New Mexico diagnosed with a malignancy from 1969 through 1982 were analyzed to determine the relationship between the age of the patient and the stage of cancer at the time of diagnosis. Three general patterns were apparent. For cancers of the bladder, breast, cervix, ovary, thyroid, and uterus, and for melanoma, there were significant linear trends for the cancers to be diagnosed at more advanced stages in older patients. For cancers of the lung, pancreas, rectum, and stomach, there were significant linear trends for cancers to be diagnosed at an earlier stage in older patients. For cancers of the colon, kidney, liver, and prostate, there were no significant linear trends in stage at diagnosis versus age of the patient.

Age Factors↗

Protein turnover in the cytoplasmic transport system within an insect ovary--a clue to the mechanism of microtubule-associated transport.

The movement of radioactively labelled polypeptides into the microtubule-associated transport channels in the ovaries of a hemipteran insect has been analysed using SDS-polyacrylamide slab gel electrophoresis and fluorography. The patterns of label suggest that the microtubules which pack the transport channels form a relatively static cytoskeleton while other components move independently from them along the channels. As well as illustrating the functional organisation of microtubule-associated transport in this system our studies of labelled proteins have also provided clues as to the mechanism of transport itself.

Animals↗

The effect of polymethylmethacrylate on bone: an experimental study.

In order to assess the response of bone to low-viscosity polymethylmethacrylate, CMW or Simplex acrylic cement was digitally packed while in a doughy state into drill holes in the proximal diaphysis in each of four long bones (humeri and tibiae) of mongrel dogs. Histological assessment was performed in areas of minimal load at the interface between the viscoelastic bone and the acrylic cement. Decalcified and undecalcified sections were evaluated and a remodeling or activity index calculated. Fluorescent labeling studies were performed in order to assess bone growth. Animals were killed at 2, 4 or 5 months. Histological analysis showed a thin connective-tissue membrane containing scattered giant cells and histiocytes at the bone-cement interface. Inflammation was not an important facet of this response. The marrow and trabecular bone were viable, except for scattered localized areas of marrow necrosis and fibrosis immediately adjacent to the cement. The bone adjacent to the cement showed a lower remodeling or activity index, fewer fluorescent bands, and smaller distances between successive bands, suggesting decreased bone formation and turnover. The etiology of these findings may include a vascular disturbance secondary to disruption of the cortical and marrow circulation, temperature effects during cement polymerization, and/or chemical effects from the acrylic monomer.

Animals↗

Binding of axonemal dynein to microtubules comprising the cytoplasmic transport system in insect ovarioles.

Dynein isolated from ciliary axonemes of Tetrahymena is shown to bind in a characteristic fashion as arms to microtubules dissected from the nutritive tubes of insect ovarioles. The microtubules in nutritive tubes are associated with the transport of cytoplasmic components along their length, and the significance of their ability to bind axonemal dynein, to the possibility that microtubule/dynein interactions are involved in microtubule-associated movements, generally, is discussed.

Adenosine Triphosphatases↗

Conserved features of eukaryotic hsp70 genes revealed by comparison with the nucleotide sequence of human hsp70.

We have determined the nucleotide sequence of the human hsp70 gene and 5' flanking region. The hsp70 gene is transcribed as an uninterrupted primary transcript of 2440 nucleotides composed of a 5' noncoding leader sequence of 212 nucleotides, a 3' noncoding region of 242 nucleotides, and a continuous open reading frame of 1986 nucleotides that encodes a protein with predicted molecular mass of 69,800 daltons. Upstream of the 5' terminus are the canonical TATAAA box, the sequence ATTGG that corresponds in the inverted orientation to the CCAAT motif, and the dyad sequence CTGGAAT/ATTCCCG that shares homology in 12 of 14 positions with the consensus transcription regulatory sequence common to Drosophila heat shock genes. Comparison of the predicted amino acid sequences of human hsp70 with the published sequences of Drosophila hsp70 and Escherichia coli dnaK reveals that human hsp70 is 73% identical to Drosophila hsp70 and 47% identical to E. coli dnaK. Surprisingly, the nucleotide sequences of the human and Drosophila genes are 72% identical and human and E. coli genes are 50% identical, which is more highly conserved than necessary given the degeneracy of the genetic code. The lack of accumulated silent nucleotide substitutions leads us to propose that there may be additional information in the nucleotide sequence of the hsp70 gene or the corresponding mRNA that precludes the maximum divergence allowed in the silent codon positions.

Amino Acid Sequence↗

A double blind study of vitamin B6 in Down's syndrome infants. Part 1--Clinical and biochemical results.

Nineteen patients with Down's syndrome participated in a double blind study of the clinical effects of pharmacological doses of vitamin B6 administration, starting under 8 weeks of age and continuing until 3 years of age. Ten patients received the vitamin and nine the placebo. In Part 1 of this study, no statistically significant differences were found between the two groups in mental age, height, weight, cranial circumference or tongue protrusion. A study of side effects conducted on a larger open population found vitamin B6 to be relatively safe when administered over long periods of time with photosensitive blisters as the major complication.

Blister↗

Structure and expression of the human gene encoding major heat shock protein HSP70.

We have cloned a human gene encoding the 70,000-dalton heat shock protein (HSP70) from a human genomic library, using the Drosophila HSP70 gene as a heterologous hybridization probe. The human recombinant clone hybridized to a 2.6-kilobase polyadenylated mRNA from HeLa cells exposed to 43 degrees C for 2 h. The 2.6-kilobase mRNA was shown to direct the translation in vitro of a 70,000-dalton protein similar in electrophoretic mobility to the HSP70 synthesized in vivo. From the analysis of S1 nuclease-resistant mRNA-DNA hybrids, the HSP70 gene appears to be transcribed as an uninterrupted mRNA of 2.3 kilobases. We show that the cloned HSP70 gene contains the sequences necessary for heat shock-induced expression by two criteria. First, hamster cells transfected with a subclone containing the HSP70 gene and flanking sequences synthesized a HSP70-like protein upon heat shock. Second, human cells transfected with a chimeric gene containing the 5' flanking sequences of the HSP70 gene and the coding sequences of the bacterial chloramphenicol acetyltransferase gene transcribed the chimeric gene upon heat shock. We show that the HSP70 mRNA transcribed in an adenovirus 5 transformed human cell line (293 cells) is identical to the HSP70 mRNA induced by heat shock.

Acetyltransferases↗

Complement- and IgE-mediated release of histamine from basophils in vitro. V. Differential effects of drugs modulating arachidonic acid metabolism.

Basophil degranulation probably plays a significant role in the pathogenesis of different hypersensitivity reactions. These cells can be stimulated to secrete lysosomal histamine in vitro in response to various secretagogues. We compared the effects of drugs, modulating arachidonic acid (AA) metabolism, on histamine release (HR) from human basophils stimulated by anti-IgE antibody or C5a anaphylatoxin. Leukocytes from normal donors were preincubated with drug for 15 min at 22 degrees C, followed by the addition of C5a or anti-IgE for 30 min at 37 degrees C. Bromophenacyl bromide, an inhibitor of AA formation by phospholipase, blocked the effects of C5a and anti-IgE (greater than or equal to 3.3 X 10(-6) M, p less than 0.05 and p less than 0.01, respectively). 3-Amino-1-(3-trifluoromethylphenyl)-2-pyrazoline hydrochloride (BW755C, greater than or equal to 3.3 X 10(-5) M) and 5, 8, 11, 14-eicosatetraynoic acid (greater than or equal to 3.3 X 10(-4) M), known inhibitors of both cyclooxygenase (COX) and lipoxygenase (LPX) pathways of AA metabolism, blocked both C5a- and anti-IgE-induced HR (p less than 0.01). Nordihydroguaiaretic acid, an inhibitor of LPX, decreased HR induced by anti-IgE (greater than or equal to 3.3 X 10(-6) M, p less than 0.01) and allergens, but reduced C5a-initiated HR only at a higher concentration (greater than or equal to 7 X 10(-5) M, p less than 0.01). Indomethacin (INDO), an inhibitor of COX, significantly reduced HR caused by C5a (greater than or equal to 3.3 X 10(-8) M, p less than 0.01) and its degradation product C5adesArg, but had no effect or caused slight enhancement of HR initiated by anti-IgE. We confirmed that INDO augments allergen-induced HR. Our findings suggest that there are basic differences in the regulation of C5a- and IgE-mediated basophil degranulation.

4,5-Dihydro-1-(3-(trifluoromethyl)phenyl)-1H-pyraz↗

Effect of cooling and warming rate on glycerolized rabbit kidneys.

Cooling and warming rates are known to be important determinants of viability for cryopreserved cells, but optimal rates have not previously been determined for any whole organ. In this study, rabbit kidneys, permeated with 2 M glycerol were cooled to -80 degrees C at four rates varying from 1 degrees C/hr to 3.1 degrees C/min and then rewarmed at four rates from 1 degrees C/hr to 4.2 degrees C/min, giving 16 experimental treatments. After gradual deglycerolization at 10 degrees C, each kidney was autografted and observed for 30 min. Assessment was by measurement of vascular resistance, immediate post-thaw lactate dehydrogenase (LDH) release, gross appearance, light- and electron microscopy, and tissue K+/Na+ ratio 30 min after transplantation. The best results were obtained after cooling at 1 degrees C/hr; warming rate had little apparent influence on the criteria used to assess function with the exception of LDH release, which indicated a preferred warming rate around 1 degrees C/min. Histological studies revealed extensive vascular damage, notably to the glomerular capillaries, that was minimized by very slow cooling. Freeze substitution, carried out on samples removed at -80 degrees C, demonstrated extensive ice formation in the interstitial space and, at the faster cooling rates, in the glomerular capillaries. Intracapillary ice formation was reduced in the kidneys cooled at 1 degrees C/hr.

Animals↗

A diagnostic approach to suspected obstructive sleep apnea in children.

Most children with obstructive sleep apnea will benefit from tonsillectomy and adenoidectomy. Although polygraphic monitoring remains the definitive diagnostic technique, we wondered if all children suspected of having OSA require such evaluation. We therefore administered a standardized questionnaire to the parents of 23 children with polygraphically proved OSA related to adenotonsillar hypertrophy, 46 age- and sex-matched normal children, and 23 children subsequently referred because of possible OSA. Significantly increased frequencies of the following symptoms were found in the OSA group compared with the control group: difficulty breathing during sleep, 96% vs 2%; apnea observed by the parents, 78% vs 5%; snoring, 96% vs 9%; restless sleep, 78% vs 23%; chronic rhinorrhea, 61% vs 11%; and mouth breathing when awake, 87% vs 18%. Using discriminant analysis, an OSA score was derived that correctly classified all control subjects and 22 of 23 patients with OSA. Considering the data from all groups, we found that (1) OSA scores greater than 3.5 were highly predictive of OSA requiring adenotonsillectomy; (2) no child with an OSA score less than -1 had OSA; and (3) in children with OSA scores between -1 and 3.5, polygraphic monitoring was required to determine the severity of sleep-related airway obstruction and the need for surgical treatment. Use of the OSA score should decrease the need for polygraphic monitoring and facilitate selection of children for tonsillectomy and adenoidectomy.

Adenoids↗