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Biomedical subjects

R F Martin

Publications and source records attributed to R F Martin.

At least 19 recordsLinked to original sources
PubMed1992

Symptomatic choledochoceles in adults. Endoscopic retrograde cholangiopancreatography recognition and management.

During a 2-year interval, we identified 10 patients with symptoms of pancreaticobiliary disorders and small choledochoceles by endoscopic retrograde cholangiopancreatography. Patients ranged from 36 to 89 years of age. Eight were female. Seven presented with recurrent, acute pancreatitis, two presented with biliary colic, and one presented with cholangitis. Dilated common bile ducts were seen in four patients, and no other biliary lesions were demonstrated in any patients. Five patients were shown to have normal gallbladders by ultrasonographic or computed tomographic criteria. Choledochoceles were identified endoscopically as a bulge above or involving the ampulla. Diagnosis was confirmed by cholangiography. All patients underwent successful unroofing of the choledochocele and sphincterotomy of the common bile duct. One pancreatic sphincterotomy was performed for pancreatic ductal obstruction. We encountered no complications. Hospital stays ranged from 1 to 4 days. Follow-up intervals ranged from 2 to 20 months. At this time, no patients have had any recurrence of symptoms, and none has required rehospitalization or surgery.

Adult↗
PubMed1992

DNA ligands as radiomodifiers: studies with minor-groove binding bibenzimidazoles.

An iodinated bibenzimidazole, iodoHoechst 33258, was previously reported to markedly sensitize DNA and cells to UV-A, exemplifying the potential of iodinated DNA ligands as radiosensitizers, a rational extension of sensitization by halogenated pyrimidines. However, unlike the latter sensitizers, iodoHoechst 33258 is not a sensitizer of ionizing radiation, presumably due to the innate radioprotective properties of the uniodinated ligand. Experiments with purified DNA show that both Hoechst 33258 and Hoechst 33342 decrease the yield the radiation-induced DNA strand breakage. The ligands bind at discrete sites in the minor groove of DNA, and analysis on DNA sequencing gels show pronounced protection at the ligand binding sites, as well as more generalized protection. The extent of protection of strand breakage on plasmid DNA and the fact that it persists in the presence of 0.5 M NaCl (which prevents low affinity ionic binding between the high affinity sites) suggests that the protective effects of bound ligand are not confined to the high affinity binding sites in the minor groove. The mechanisms of this generalized protection is unknown, but there is some evidence indicating that the H-atom donation from the ligand may account for the site-specific protection. The extent of protection is much diminished, but still evident, in the presence of 100 mM mannitol, a known hydroxyl radical scavenger, indicating that some of the protective effects might relate to DNA damage mediated by direct action. Further evaluation of the mechanisms of protection should enable development of both more active radioprotectors and, by elimination of the radioprotective features from halogenated DNA ligands, more effective radiosensitizers.

Benzimidazoles↗
PubMed1992

DNA ligands as radioprotectors: molecular studies with Hoechst 33342 and Hoechst 33258.

Following earlier reports of radioprotection of cells by Hoechst 33342, we have investigated radioprotection of isolated DNA by the minor groove binders Hoechst 33258 and Hoechst 33342. Analysis of radiation-induced single strand breakage in plasmid DNA (pBR322) showed concentration-dependant protection, up to a dose-modifying factor of 9.3 for 25 microM Hoechst 33258, at which the ligand: bp ratio was 0.67. Since the ligands bind at discrete sites along DNA, sequencing gel analysis was used to investigate the radioprotective effects of the ligands both at and between the ligand-binding sites. These experiments showed that although protection was more pronounced at the binding sites, there was also some reduction in strand-breakage between binding sites. Detailed analysis at a particular site, the EcoR1 site in a 3'-32P-endlabelled 100bp restriction fragment from pBR322, showed that protection was most pronounced at the 'inner T': GAATTC. Irradiation of a synthetic oligodeoxynucleotide containing a single ligand-binding site, and labelled at the 5'-end, gave the expected doublet bands in high resolution gels, corresponding to fragments with 3'-phosphoryl- and 3'-phosphorylglycollate terminii. In the Hoechst 33258-protected sample, the 3'-phosphorylglycollate band was preferentially suppressed within the binding site. These results, together with published crystal structure data for a Hoechst 33258/dodecamer complex, suggest that the site-specific radioprotection may be due to H-atom donation from the benzimidazole NH groups in the ligand to radiation-induced radicals on 4'-deoxyribosyl carbons. In contrast to the experiments with purified DNA, in which the two ligands yielded similar results, Hoechst 33342 was a much more active radioprotector in experiments with intact cells. For 20 microM Hoechst 33342, the dose-modifying factor was 1.7 at 1% survival and 1.3 at 10% survival, whereas the same level of Hoechst 33258 yielded barely detectable protection, perhaps due to a demonstrably lower cellular uptake. Presumably the radioprotection of cells by Hoechst 33342 is due to suppression of DNA strand breakage, and further investigation of the protection mechanism(s) should enable development of improved radioprotectors.

Base Sequence↗
PubMed1991

Necrotizing enterocolitis in a neonatal piglet model.

The aim of this study was to develop an animal model for necrotizing enterocolitis (NEC). Twenty-five neonatal Hanford minipigs had carotid artery and external jugular vein catheters and rectal Clinical Tonomitors placed under anesthesia. Experimental animals were subjected to a hypoxic insult (50% reduction in baseline PaO2 for 30 minutes) and hypothermic stress (core temperature reduced to 35 degrees C for 30 minutes). Regular oral diet was resumed and the survivors were euthanized 3 to 4 days later. All animals underwent necropsy with gross and histopathological evaluation of the entire bowel. Of 22 experimental animals, 14 survived (64%) and 8 (36%) died of pulmonary hemorrhage. Of the 14 survivors, 8 (57%) had gross and microscopic evidence of NEC. Six of the total 25 animals (24%) sustained rectal perforations from the tonometer. Of 3 control animals, one died of pulmonary hemorrhage and the two survivors had normal intestine. This model successfully produced gross and histological evidence of NEC. The tonometer shows promise as a predictor of NEC provided technical modifications can reduce the complication rate.

Animals↗
PubMed1991

Blunt trauma to the carotid arteries.

Blunt carotid artery trauma is an uncommon but potentially dangerous clinical entity. We report eight patients from a 10-year interval who sustained blunt injuries to the carotid arteries. Six of eight patients suffered a hyperextension injury or had a cervical spine fracture or both. Arteriography revealed four arterial dissections and four thrombotic occlusions. Two asymptomatic common carotid artery occlusions and one dissection with transient ischemic attacks had successful arterial reconstructions. Five patients were treated nonoperatively: three internal carotid artery dissections with minor or no neurologic deficit; one asymptomatic thrombosis; and one internal carotid artery thrombosis with a major fixed neurologic deficit that did not improve. No patient died, and seven of eight made a complete neurologic recovery or remained asymptomatic. The diagnosis of blunt carotid artery injuries should be suspected in patients with neck hyperextension injuries or with cervical spine fractures as well as in patients with neurologic deficits not explained by intracranial trauma. Duplex scanning may be a useful noninvasive study. Surgery is indicated for selected patients with accessible lesions who have minor or no neurologic deficits. Asymptomatic patients with small intimal flaps or dissections may be successfully treated nonoperatively.

Adolescent↗
PubMed1990

Radiation sensitization by an iodine-labelled DNA ligand.

An iodinated DNA ligand, iodo Hoechst 33258, which binds in the minor groove of DNA, enhances DNA strand breakage and cell killing by UV-A irradiation. The sites of UV-induced strand breaks reflect the known sequence specificity of the ligand.

Animals↗
PubMed1990

The selenite-exchangeable metabolic pool in humans: a new concept for the assessment of selenium status.

An in vivo isotope-dilution approach is considered for assessment of selenium status in human subjects. The approach depends upon the dilution of a single dose of the stable isotope 74SeO3(2-) in the selenite-exchangeable metabolic pool. Data from six metabolic protocols, conducted with healthy North American males, are presented in order to analyze characteristics of this pool. Pool size (WSe-EMP) correlated positively with daily selenium intake in subjects consuming diets of known and variable selenium content. When subjects were given a selenium-adequate or -restricted diet for 30 d, WSe-EMP,7d decreased from 4.49 +/- 0.28 to 3.76 +/- 0.22 mg (p less than 0.05). The corresponding 24-h urinary selenium concentration dropped from 0.556 +/- 0.035 to 0.341 +/- 0.058 mumol/d (means +/- 1 SEM). Route of administration (iv vs po) had no apparent effect on WSe-EMP. In subjects of similar selenium status, the WSe-EMP was reproducible within the expected uncertainties of the method. This approach may be suitable for assessment of selenium status for a wide range of chronic intakes.

Adult↗
PubMed1990

Pokeweed mitogen-stimulated human lymphocytes fused to LICR-2 (HMY2) generate human-human hybridomas producing monoclonal IgG antibodies reactive to human breast carcinoma and malignant melanoma.

Human-human hybridomas were generated using pokeweed mitogen-stimulated lymphocytes from the regional lymph nodes of cancer patients by fusion to the LICR-2 human myeloma cell line. A total of 35 fusions, using the regional lymph node lymphocytes of cancer patients, resulted in hybrid growth in 23% of wells plated with 21 IgG ELISA positive clones, 6 of which have maintained stable human monoclonal antibody production. Mononuclear cells were separated on Ficoll-Paque and grown for 3-4 days in 1% pokeweed mitogen and fused to the LICR-2 human myeloma cell line. Human-human hybridoma producing membrane reactive IgG antibodies have been isolated and react to the following cancers: breast; melanoma. Twenty-seven fusions from 8 breast carcinoma patients resulted in 13 ELISA positive IgGs, 3 of which were stable after cloning. A total of 5,071 wells were plated after polyethylene glycol fusion with resultant hybrid growth in 1210 wells (24% hybrid growth) after hypoxanthine-aminopterin-thymidine selection. In 8 fusions using regional lymph node lymphocytes of other types of cancer, including 6 fusions using lymphocytes from malignant melanoma patients, there were 1,580 wells plated with positive growth in 20% of the wells (311 wells). Of these, 8 clones were ELISA positive and 3 stable clones all producing IgG anti-melanoma antibody were isolated. The overall hybrid frequency was 43 x 10(-7) fused lymphocytes (39 x 10(-7) non-breast and 45 x 10(-7) breast). A total of 21 IgG-producing clones were identified to crude membranes of allogeneic tumor cell lines and stable antibody production was achieved for 6 (29% stable clones).(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Monoclonal↗
PubMed1989

The degree of ultraviolet light damage to DNA containing iododeoxyuridine or bromodeoxyuridine is dependent on the DNA sequence.

The sequence selectivity of 300 nm ultraviolet light damage to DNA containing bromodeoxyuridine or iododeoxyuridine was examined on DNA sequencing gels. This was accomplished using a system where an M13 template was employed to direct synthesis of DNA in which thymidine was fully substituted with bromodeoxyuridine or iododeoxyuridine. The sites of damage corresponded to the positions of analogue incorporation. The extent of damage varied considerably at different sites of cleavage and ranged from the undetectable to over fifteen times the limit of detection (as assessed by laser densitometer scans). Strong damage sites had the "consensus" sequence CTT while sites of no detectable damage had the "consensus" sequence GTR. Bromodeoxyuridine and iododeoxyuridine had the same sites of damage although the extent of damage varied at different sites and bromodeoxyuridine damage was slightly greater than iododeoxyuridine. DNA containing thymidine was not damaged to any detectable level in this system with 300 nm ultraviolet light. The use of three closely related DNA sequences as targets for damage confirmed that (1) the sites of analogue incorporation are the cause of ultraviolet damage; and (2) that the neighbouring DNA sequence is an important parameter in determining the extent of damage. It is proposed that the microstructure of DNA--in particular the distance between the 5-carbon of the pyrimidine base (which is attached to the halogen) and hydrogen on the 2' carbon of the 5'-deoxyribose--ultimately determines the degree of cleavage with large distances giving a small degree of damage and smaller distances a large degree of damage.

Adenosine↗
PubMed1989

Nociceptive responses of trigeminal neurons in SII-7b cortex of awake monkeys.

A cluster of trigeminal nociceptive neurons was located in the lateral sulcus on the upper bank of the frontoparietal operculum in a region bordering between cortical areas SII and 7b. These neurons were isolated in cortical cell layers IV and V-VI. All nociceptive neurons responded exclusively to noxious mechanical stimulation of cutaneous receptive fields on the face/head or intraoral tissue. Sustained noxious mechanical stimulation elicited slowly adapting responses that accurately encoded the duration of the stimulation. Prolonged discharges following removal of noxious stimulation were not observed. These nociceptive specific neurons poorly encoded graded noxious stimuli. Trigeminal somatosensory neurons within and surrounding the SII-7b cluster were not topographically organized according to divisions of the trigeminal nerve, laterality of receptive fields, or division of face/head and intraoral receptive fields. The thalamocortical, corticocortical and indirect corticolimbic connectivities of SII and area 7b and the possible role of SII-7b nociceptive neurons in learning, memory and avoidance behaviors are discussed.

Action Potentials↗
PubMed1989

Metaphyseal impaction fractures in acute lymphoblastic leukemia.

Patients with acute lymphatic leukaemia frequently are osteoporotic. A small subset of these develop disabling metaphyseal transverse fractures, usually bilateral and in the lower limb. These impaction fractures have a characteristic appearance and develop in recently laid down bone. They may develop ab initio or during therapy. Magnesium deficiency is found in these patients.

Child↗
PubMed1989

Experimental selenium restriction in healthy adult humans: changes in selenium metabolism studied with stable-isotope methodology.

Mechanisms responsible for selenium homeostasis were investigated in healthy adult men receiving diets adequate or low in Se (eight subjects per group). The appearance of a stable isotope of Se, 74Se, in plasma, urine, and feces was measured after oral administration of 74Se-selenite. One group received a restricted level of Se (18 +/- 1 micrograms/d) for 30 d, which resulted in a decrease in urinary, fecal, and plasma Se content compared with the group that consumed 119 +/- 1 micrograms/d. Low Se intake also resulted in decreased urinary 74Se excretion (27.2 +/- 1.4% vs 32.5 +/- 2.3% of the absorbed dose for the adequate intake), increased body retention of 74Se (74.8 +/- 3.1% vs 67.6 +/- 3.8% of the absorbed dose for the adequate group), and a contracted selenite-exchangeable metabolic pool (Se-EMP) (9782 micrograms for adequate Se and 6314 micrograms for the low-Se group; p less than or equal to 0.05). Measurement of Se-EMP may provide an additional and sensitive approach for assessing Se nutriture in human subjects.

Absorption↗
PubMed1989

Ascorbic acid-selenite interactions in humans studied with an oral dose of 74SeO3(2-).

The interaction between dietary ascorbic acid at extremes of ascorbic acid intake and selenium in young adult male humans was investigated with a stable-isotope approach using 74Se-selenite. Measurements were made of 74Se in plasma, urine, and feces with neutron-activation analysis after oral administration of 74SeO3(2-). Urine excretion and total body retention of isotope and the selenite-exchangeable metabolic pool (Se-EMP) were calculated. Limiting dietary ascorbic acid to about 20 mg/d appeared to reduce the time-related retention of absorbed selenite and the size of Se-EMP. Compared with a diet providing 1 g ascorbic acid/d the low ascorbic acid intake was associated with a lower fractional absorption of the isotope, a reduced retention of the label, and a smaller Se-EMP. These data and those previously obtained in subjects with more usual ascorbic acid intakes point to a possible important role for ascorbic acid in the maintenance of Se homeostasis.

Absorption↗
PubMed1988

The sequence specificity of bleomycin damage in three cloned DNA sequences that differ by a small number of base substitutions.

The DNA sequence specificity of the cancer chemotherapeutic agent, bleomycin, has been investigated in three clones of human alpha RI-DNA. The three 340-base pair alpha RI-DNA sequences were almost identical in their nucleotide sequence enabling the study of subtle effects of base substitutions on bleomycin cleavage. By utilizing densitometer scanning and statistical analysis of the degree of bleomycin DNA cleavage, we found 17 significant differences between the three DNA sequences. Eleven of these differences could be attributed to base substitutions close to the dinucleotide cleavage site. However, six of the differences were at positions two or more base pairs from the base substitution sites. The significant differences were up to 12 base pairs from base substitutions. It is proposed that these long range effects are due to base substitutions causing microvariation in the DNA structure to which bleomycin cleavage is sensitive.

Base Sequence↗
PubMed1988

Sequence specificity of 125I-labelled Hoechst 33258 damage in six closely related DNA sequences.

The sequence selectivity of [125I]Hoechst 33258 in six 340 base-pair DNA sequences has been investigated. [125I]Hoechst 33258, which is a bis-benzimidazole and binds to the minor groove of B-DNA, preferentially binds to A + T-rich regions of DNA. Six out of nine strong binding sites contained four or more consecutive A.T base-pairs, while the other three strong binding sites were AAGGATT, TATAGAAA (the peak of damage was in the run of 3 A residues) and AAA. One of the six weak binding sites had five consecutive A.T base-pairs, two of the weak binding sites had three, and three did not have any. In addition to genomic 340 base-pair alpha RI-DNA (which is a tandem repeat in human cells), five 340 base-pair alpha RI-DNA clones were generated that differed from the genomic "consensus" sequence by a number of random base alterations. The effect of these base changes on the sequence specificity of [125I]Hoechst 33258 damage indicated that of the base changes that interrupted 14 binding sites, six decreased and eight did not change the extent of damage, while two sites changed position. Of the base alterations that augmented 17 binding sites, five increased, two decreased and ten did not alter the degree of cleavage, while ten sites changed position. It was concluded from the data that, while runs of consecutive A.T base-pairs was the most important parameter that determines [125I]Hoechst 33258 binding, other factors including position in the DNA sequence, nearest neighbour and long-range interactions were also important.

Autoradiography↗
PubMed1988

Sequence specificity of 125I-labelled Hoechst 33258 in intact human cells.

Using polyacrylamide/urea DNA sequencing gels, the DNA sequence selectivity of 125I-labelled Hoechst 33258 damage has been determined in intact human cells to the exact base-pair. This was accomplished using a novel procedure with human alpha RI-DNA as the target DNA sequence. In this procedure, after size fractionation, the alpha RI-DNA is selectively purified by hybridization to a single-stranded M13 clone containing an alpha RI-DNA insert. The sequence specificity of [125I]Hoechst 33258 was indistinguishable in intact cells from purified high molecular weight DNA; and this is surprising considering the more complex environment of DNA in the nucleus where DNA is bound to nucleosomes and other DNA binding proteins. The ligand preferentially binds to DNA sequences which have four or more consecutive A.T base-pairs. The extent of damage was measured with a densitometer and, relative to the damage hotspot at base-pair 94, the extent of damage was similar in both purified high molecular weight DNA and intact cells. [125I]Hoechst 33258 causes only double-strand breaks, since single-strand breaks or base damage were not detected. These experiments represent the first occasion that the sequence specificity of a DNA damaging agent, which causes only double-strand breaks, has been determined to the exact base-pair in intact cells.

Autoradiography↗
PubMed1988

Induction of double-strand breaks following neutron capture by DNA-bound 157Gd.

Irradiation of plasmid DNA/Gd3+ mixtures with thermal neutrons induces DNA double-strand breaks (dsb). However, the extent of breakage is markedly reduced by sequestering the Gd3+ from DNA by addition of EDTA. Since the 157Gd neutron capture event involves some internal conversion, we suggest that the DNA dsb induction results from Auger electron emission.

DNA↗