PubMed Health⌕ Search

Biomedical subjects

D A Gilbert

Publications and source records attributed to D A Gilbert.

At least 19 recordsLinked to original sources
PubMed1992

Molecular phylogenetic inference from saber-toothed cat fossils of Rancho La Brea.

A method for the successful extraction of sequestered cellular DNA from 14,000-year-old fossil bones was developed and applied to asphalt-preserved specimens of the extinct saber-toothed cat, Smilodon fatalis. Two distinct gene segments, the mitochondrial gene for 12S rRNA and nuclear FLA-I (the feline class I major histocompatibility complex gene), from three different individual fossil specimens were cloned and sequenced after PCR amplification. Comparison of fossil-derived DNA sequences to homologous regions in 15 living carnivorous species, including 9 species of Felidae and 6 nonfelids, affirmed the phylogenetic placement of Smilodon within the modern radiation of Felidae distinct from the Miocene paleofelid (Nimravidae) saber-toothed "cat" species. These results raise the prospect of obtaining genetically informative DNA from preserved bones of extinct fossil species, particularly among the 2 million specimens excavated from the asphaltic sediments at Rancho La Brea in metropolitan Los Angeles.

Animals↗
PubMed1992

Estimation of heterozygosity for single-probe multilocus DNA fingerprints.

In spite of the increasing application of DNA fingerprinting to natural populations and to the genetic identification of humans, explicit methods for estimation of basic population genetic parameters from DNA fingerprinting data have not been developed. Contributing to this omission is the inability to determine, for multilocus fingerprinting probes, relatively important genetic information, such as the number of loci, the number of alleles, and the distribution of these alleles into specific loci. One of the most useful genetic parameters that could be derived from such data would be the average heterozygosity, which has traditionally been employed to measure the level of genetic variation within populations and to compare genetic variation among different loci. We derive here explicit formulas for both the estimation of average heterozygosity at multiple hypervariable loci and a maximum value for this estimate. These estimates are based upon the DNA restriction-pattern matrices that are typical for fingerprinting studies of humans and natural populations. For several empirical data sets from our laboratory, estimates of average and maximal heterozygosity are shown to be relatively close to each other. Furthermore, variances of these statistics based on simulation studies are relatively small. These observations, as well as consideration of the effect of missing alleles and alternate numbers of loci, suggest that the average heterozygosity can be accurately estimated using phenotypic DNA fingerprint patterns, because this parameter is relatively insensitive to the lack of certain genetic information.

Animals↗
PubMed1992

Microsurgical forearm "cricket bat-transformer" phalloplasty.

Presently, the donor flap of choice for microsurgical phallic reconstruction is the radial forearm flap. The success of several different design modifications confirms the reliability of the radial and ulnar forearm flaps. Farrow et al. described their experience with the "cricket bat" concept in 1980. To the previous "cricket bat" design, we now wish to add modifications. These modifications utilize longitudinal and transverse rotations of the linear forearm tissues to create a phallus--much like the transformation of a toy robot into a truck. Deepithelialized flaps and a full-thickness skin graft coronoplasty complete glans reconstruction. The "cricket bat-transformer" flap appears to produce the most predictable results in subtotal phallic reconstructions and phallic constructions in the pediatric and transgender patient groups.

Adult↗
PubMed1991

The surgical treatment of chordee without hypospadias in men.

During a 2-year period we treated 26 young men for chordee without hypospadias. Many of these patients had straight erections as children but a ventral curvature developed as they achieved puberty. We describe the anatomical findings and discuss the possible cause for the development of this anomaly. Surgical therapy begins with a circumcising incision and reflection of the skin to expose the shaft of the penis. The corpus spongiosum containing the urethra was mobilized by resecting the dysgenetic tissue in the dartos and Buck's fascia layers. In 1 patient this dissection was sufficient to straighten the penis but in the remaining 25 the penis was not straight. In those patients we mobilized the dorsal bundle of vessels and nerves, and removed 1 or several ellipses of tunica albuginea to equalize the lengths of the ventral and dorsal aspects of the corpora cavernosa. The corpus spongiosum usually is elastic and the curve almost never is caused by shortness of the urethra, which stretches to fit the straightened penis. In 24 of the 26 patients the curvature was resolved with 1 operation, while 2 needed a second procedure.

Adolescent↗
PubMed1991

In vitro inhibition of hepatitis B virus replication by 2',3'-dideoxyguanosine, 2',3'-dideoxyinosine, and 3'-azido-2',3'-dideoxythymidine in 2.2.15 (PR) cells.

Hep G2-derived hepatoblastoma cells (2.2.15), which actively produce hepatitis B virus (HBV), were cultured in the presence of 2',3'-dideoxyguanosine (ddG), 2',3'-dideoxyinosine, or 3'-azido-2',3'-dideoxythymidine (AZT). ddG was the most potent agent. It diminished viral replication by up to 95%, as assessed by the amount of episomal HBV DNA, without impairing cellular growth. AZT was the least effective against HBV. Northern blot analysis revealed no apparent difference in the pregenomic viral RNA profile, suggesting that these dideoxynucleosides suppress reverse transcription in the replicative cycle of HBV. The effect of varying the time of drug exposure showed that these agents can suppress HBV replication even when added late in culture. HBV replication in another 2.2.15 cell population of the same lineage was affected by ddG differently, which may enable the investigation of phenotypic or genetic alterations during culture. The present data suggest that some 2',3'-dideoxynucleosides can exert a potent antiviral activity against HBV in vitro, at least under certain circumstances, although the data do not prove that any of these agents have utility in patients with hepatitis.

Blotting, Southern↗
PubMed1990

Oscillations in cell morphology and redox state.

Fluctuations in the intensity of light scattered and absorbed by cells in suspension have been analysed by smoothing, periodogram and power spectrum methods to reveal oscillations attributed to changes in cell morphology and the redox state of NADH and FAD (periods 10 s to 30 min). The rhythms are themselves periodically modulated in amplitude at a similar frequency and exhibit burst characteristics. The low frequency scatter dynamics are provisionally attributed to oscillations in gross morphology and the high frequency variation to changes at the cell surface. Agents, such as insulin and transferrin, affect the dynamics. The scatter results suggest that rhythmic changes in cell morphology associated with locomotion are largely inherent in the cell and not due to periodic attachment and detachment from a surface.

Animals↗
PubMed1990

Genetic fingerprinting reflects population differentiation in the California Channel Island fox.

Restriction fragment profiles generated by hybridization of hypervariable minisatellite DNA probes have been used for paternity analysis but not for comparisons at the level of populations, because the profiles are thought to evolve too rapidly to be informative over large time intervals. But in small isolated populations, the fixation of restriction-fragment polymorphisms can outpace the generation of fragment-length variability through recombination. Here we report on an analysis of DNA fingerprints of the California Channel Island fox (Urocyon littoralis). These foxes comprise an island dwarf species found only on six of the Channel Islands off the coast of southern California. Variability of restriction-fragment profiles within fox populations, as indicated by the average percentage difference (APD), varied widely among the islands, from 0.0% (no variation) to 25.3%. The APDs between populations were considerably greater (43.8% to 84.4%). In addition, foxes on each island can be distinguished by the presence of diagnostic restriction fragments. Maximum parsimony and phenetic trees relating foxes from different islands are consistent with the archaeozoological and geological record. Therefore, in small populations of genetically isolated mammals, differences among hypervariable restriction-fragment profiles can be used to estimate relative genetic variability and to reconstruct the evolutionary relationships of natural populations.

Animals↗
PubMed1990

Application of DNA fingerprints for cell-line individualization.

DNA fingerprints of 46 human cell lines were derived using minisatellite probes for hypervariable genetic loci. The incidence of 121 HaeIII DNA fragments among 33 cell lines derived from unrelated individuals was used to estimate allelic and genotypic frequencies for each fragment and for composite individual DNA fingerprints. We present a quantitative estimate of the extent of genetic difference between individuals, an estimate based on the percentage of restriction fragments at which they differ. The average percent difference (APD) among pairwise combinations from the population of 33 unrelated cell lines was 76.9%, compared with the APD in band sharing among cell lines derived from the same individual (less than or equal to 1.2%). Included in this survey were nine additional cell lines previously implicated as HeLa cell derivatives, and these lines were clearly confirmed as such by DNA fingerprints (APD less than or equal to 0.6%). On the basis of fragment frequencies in the tested cell line population, a simple genetic model was developed to estimate the frequencies of each DNA fingerprint in the population. The median incidence was 2.9 X 10(-17), and the range was 2.4 X 10(-21) to 6.6 X 10(-15). This value approximates the probability that a second cell line selected at random from unrelated individuals will match a given DNA fingerprint. Related calculations address the chance that any two DNA fingerprints would be identical among a large group of cell lines. This estimate is still very slight; for example, the chance of two or more common DNA fingerprints among 1 million distinct individuals is less than .001. The procedure provides a straightforward, easily interpreted, and statistically robust method for identification and individualization of human cells.

Alleles↗
PubMed1989

Temporal changes in phosphoamino acid phosphatase activities in murine erythroleukaemic cells.

1. The activities of phosphotyrosine, phosphothreonine and phosphoserine phosphatases were measured at various time periods in Friend murine erythroleukaemic (MEL) cells. 2. Effects of DMSO (dimethyl sulphoxide) and HMBA (hexamethylene bisacetamide), inducers of differentiation, were examined. 3. The activities of all three enzymes showed cyclic variation when measured on a daily basis over a period of several days; this was also the case when phosphothreonine phosphatase was determined in cells treated with DMSO or HMBA and when phosphoserine phosphatase was assayed after stimulation of the cells with HMBA. 4. Evidence for rhythmic variation in phosphotyrosine phosphatase activities was also obtained when the activities were determined at hourly intervals both in control cells and those treated with HMBA. 5. The time-dependent changes observed could be significant in that control of dephosphorylation may possibly be achieved by altering the rhythms.

Animals↗
PubMed1989

Isolation and characterization of collagen in Peyronie's disease.

Peyronie's disease is characterized histologically by excessive collagen deposition in the lesion. We examined the collagen types in Peyronie's disease plaque tissues compared to unaffected tissues from the same patient, other control tissues, and Dupuytren's contracture. Gel electrophoresis of pepsin-solubilized collagen demonstrated the presence of type I collagen and an increased content of type III collagen in plaque tissue. Increased type III collagen was detected in apparently normal tissue adjacent to the plaque and in Dupuytren's lesion, confirming previous findings. Although the cause of excessive collagen accumulation of Peyronie's disease is unknown, the results suggest an imbalance in the regulation of extracellular matrix production leading to pathologic fibrosis.

Collagen↗
PubMed1989

The vascularized fascia of the scalp.

The fascial layers of the temporal and occipital regions of the scalp were examined in 11 fresh cadavers. In the temporal region, three independently vascularized layers were isolated, all of which could be elevated on a single superficial temporal artery, but separated to remain independently vascularized from specific branches of this parent trunk. In the occipital area, the occipital vessels could be dissected to yield a long pedicle for an independent, fascial territory that could be transposed locally or elevated as a free flap and that will, in all likelihood, carry vascularized occipital bone. Realized and as yet unrealized uses of these ultrathin vascularized tissues remain boundless. Three representative cases are presented.

Adult↗
PubMed1989

Management of amputation injuries of the male genitalia.

Modern tissue transfer techniques have drastically modified our approach to reconstructive surgery in general. Certainly, the management of traumatic genital amputation is no exception. Although the unique vascular properties of the penis have allowed for astonishingly good results in a number of cases of genital amputation injuries, it is certain that microreplantation procedures make for uniformly good results with a minimum of postoperative complications. After microreplantation, the patient is left with a penis that is cosmetically normal and functionally nearly normal or undetectably abnormal. Although microreplantation offers the best results, certainly in the case of penile amputation, if microreplantation technology is not available, the older corporal reattachment techniques should be offered.

Amputation, Traumatic↗
PubMed1988

Chromosomal mapping of lysosomal enzyme structural genes in the domestic cat.

A panel of 42 rodent x cat somatic cell hybrids has been used to assign seven structural genes for lysosomal enzymes to specific chromosomes in the domestic cat. The assignments include alpha-glucosidase (GANAB) to chromosome D1, alpha-galactosidase (GLA) to the X chromosome, beta-galactosidase 1 (GLB1) to chromosome B3, beta-glucuronidase (GUSB) to chromosome E3, alpha-mannosidase A (MANA) to chromosome B3, alpha-L-fucosidase (FUCA) to chromosome C1, and hexosaminidase A (HEXA) to chromosome B3. In all cases, the feline lysosomal enzyme genes were located in linkage groups which were syntenic with their homologous positions in the human gene map. These assignments expand the genetic map of the cat and reaffirm the extensive syntenic homology between the chromosome maps of man and cat.

Animals↗
PubMed1988

Phallic reinnervation via the pudendal nerve.

Total phallic reconstruction presents the genitourinary reconstructive surgeon with one of the most difficult surgical challenges. The development of microsurgical techniques and free tissue transfers have advanced phallic reconstruction by reducing the number of surgical procedures and by allowing more selectivity in choosing the best innervated donor tissue. During the last 5 years 16 patients underwent total phallic reconstruction using free tissue transfers from distant donor sites. The pudendal nerve was coapted routinely to the major sensory nerves of the donor free flap. The most accurate objective baseline parameters of penile sensibility are pressure and vibratory thresholds, and electrically evoked potentials. We examined 30 normal subjects and 7 patients at least 1 year postoperatively for penile (phallic) sensibility. A pressure aesthesiometer, a biothesiometer and electrodiagnostic studies were used for testing. The 7 postoperative patients (in all of whom the pudendal nerve was incorporated into the reconstruction) had an encouraging return of tactile and erogenous sensibility compared to normal subjects. This is a promising advance in phallic reconstruction.

Adolescent↗