Search PubMed⌕ Search

Biomedical subjects

J Bowles

Publications and source records attributed to J Bowles.

At least 19 recordsLinked to original sources
PubMed2001

Regulation of male sexual development by Sry and Sox9.

Sry, a gene from the Y chromosome, is known to initiate testis formation and subsequent male differentiation in mammals. A related gene, Sox9, also plays a critical role in testis determination, possibly in all vertebrates. A number of models have been presented regarding the molecular modes of action of these two genes. However, details regarding their regulation, regulatory target genes, and interacting protein factors and co-factors have not been established with any certainty. In this review, we examine new evidence and re-examine existing evidence bearing on these issues, in an effort to build up an integrative model of the network of gene activity centred around Sry and Sox9. J. Exp. Zool. 290:463-474, 2001.

Animals↗
PubMed2001

Searching for missing pieces of the sex-determination puzzle.

Little is known of the mechanisms whereby the mammalian indifferent gonad develops into a testis or ovary. In XY individuals, Sry, the mammalian testis-determining gene, is expressed in the pre-Sertoli cells, which then differentiate into Sertoli cells. Other cell types, which include the germ cells, the steroidogenic cells and the connective tissue cells, must then be instructed to develop in a male-specific manner. Although some genes involved in sex-determination and differentiation processes have been identified, we know little of how they interact and cooperate to orchestrate the development of a testis or ovary. We have initiated an expression-screening program designed to identify additional genes, known or novel, which play a role in these processes. This approach is based on our belief that many of the genes we seek will be expressed in a sex-specific manner during the period of sex-determination and differentiation. Most of the genes identified previously are transcription factors and so we aim, in particular, to find genes involved in cell-to-cell communication, signal transduction, and transcriptional regulation, downstream of the differentiation of Sertoli cells. We have used a suppression subtractive-hybridization method to generate male- and female-enriched probes and libraries. Clones are validated as being sex-specific in their expression patterns by array screening and in situ hybridization. Here we report on our progress to date and the general applicability of the approach for studies in other systems. J. Exp. Zool. 290:517-522, 2001.

Animals↗
PubMed2001

New clues to the puzzle of mammalian sex determination.

WT1 and SOX9 are transcription factors with critical roles in mammalian sex determination and gonadal development. Recent studies in vivo clarify the roles of two alternative splice isoforms of WT1, and demonstrate that SOX9 can induce male sex determination.

Animals↗
PubMed2001

Ex vivo sentinel node mapping in carcinoma of the colon and rectum.

OBJECTIVE: Increasing evidence supports that the sentinel node (SN) is at greatest risk for harboring metastatic disease. This study describes a novel technique to identify the SN in colorectal carcinoma. METHODS: Within 30 minutes of resection, colorectal specimens were injected submucosally with isosulfan blue in four quadrants. Blue lymphatic channels were identified in the mesentery and followed to the blue-stained SN(s), which were then harvested. The specimen was fixed in formalin and subsequently analyzed in the usual fashion. Blue-stained nodes that were negative by hematoxylin and eosin staining were further analyzed by immunohistochemical staining. RESULTS: During a 6-month period, 26 patients with adenocarcinoma of the colon and rectum undergoing routine resection were studied. There were 18 men and 8 women ranging in age from 29 to 86 years (median 66). Blue-stained SNs were identified in 24 of 26 specimens. The mean number of SNs identified per patient was 2.8 +/- 1.6. Seventy-three SNs were identified from a total of 479 lymph nodes harvested. The mean number of nodes identified per patient was 18.4 +/-7. A total of 67 lymph nodes in 12 patients were identified by hematoxylin and eosin staining to have evidence of metastatic disease. Fourteen (20%) of these nodes in six patients were stained blue. However, with immunohistochemical staining, only one blue node did not have evidence of metastatic tumor in a lymphatic basin with tumor present. Four patients (29%) whose lymphatic basins were negative by hematoxylin and eosin staining were upstaged by immunohistochemical staining of the SN. CONCLUSIONS: Ex vivo mapping of the colon and rectum is technically feasible and may provide a useful approach to the ultrastaging of colorectal carcinoma.

Adenocarcinoma↗
PubMed2000

Phylogeny of the SOX family of developmental transcription factors based on sequence and structural indicators.

Members of the SOX family of transcription factors are found throughout the animal kingdom, are characterized by the presence of a DNA-binding HMG domain, and are involved in a diverse range of developmental processes. Previous attempts to group SOX genes and deduce their structural, functional, and evolutionary relationships have relied largely on complete or partial HMG box sequence of a limited number of genes. In this study, we have used complete HMG domain sequence, full-length protein structure, and gene organization data to study the pattern of evolution within the family. For the first time, a substantial number of invertebrate SOX sequences have been included in the analysis. We find support for subdivision of the family into groups A-H, as has been suggested in some previous studies, and for the assignment of two new groups, I and J. For vertebrate genes, it appears that relatedness as suggested by HMG domain sequence is congruent with relatedness as indicated by overall structure of the full-length protein and intron-exon structure of the genes. Most of the SOX groups identified in vertebrates were represented by a single SOX sequence in each invertebrate species studied. We have named anonymous sequences and, where appropriate, have suggested systematic names for some previously identified sequences. In addition, we identify an HMG domain signature motif which may be considered representative of the SOX family. Based on our data, we propose a robust phylogeny of SOX genes that reflects their evolutionary history in metazoans.

Amino Acid Sequence↗
PubMed2000

Characterisation of Crim1 expression in the developing mouse urogenital tract reveals a sexually dimorphic gonadal expression pattern.

The Crim1 gene encodes a putative transmembrane protein with an IGF-binding protein motif and multiple chordin-like cysteine-rich repeats. In chordin, such repeats are responsible for its dorsalising activity and for binding to bone morphogenic proteins (BMPs). Crim1 displays a dynamic expression pattern in a variety of developing organs, including the CNS and the lens. We have undertaken a detailed expression pattern analysis of Crim1 in the developing mouse urogenital system. During metanephric development, Crim1 showed expression both in the ureteric tree, the early condensing mesenchyme and distal comma-shaped bodies. As the nephron elongates, Crim1 becomes expressed in the proximal end of the S-shaped bodies. Crim1 also displays a striking male-specific expression pattern in the fetal gonads, its expression strongest in the Sertoli cells of the developing testis.

Animals↗
PubMed2000

Shattered: Medawar's test tubes and their enduring legacy of chaos.

Medawar's 1952 paper 'An Unsolved Problem of Biology' underlies most subsequent theoretical work regarding the evolution of aging; it concludes that aging is accidental and could not have evolved; this prevents reconciling the growing body of evidence suggesting the existence of multiple, evolved, aging systems. The paper features a well-known thought experiment using test tubes to show why aging could not evolve. Medawar assumes that constant, random, breakage sufficiently represents lethal forces of nature; however, famine, drought, predation, disease, and accidents each uniquely affect populations. Predation is the only evolving force that continually invents new ways to kill members of the prey populations; thus all prey defenses to predation will eventually be defeated. Defenses to non-evolving or non-obligate lethal forces, however, should quickly evolve. Thus unevolving, identical test tubes cannot adequately represent biological populations. The example also ignores population booms and busts which often occur in nature. By ignoring these issues, Medawar examines only one population age distribution skewed towards younger individuals in predator-dominated environments while ignoring predator-free populations skewed towards older individuals after population crashes. Further, Medawar's test tubes lack meaningful competition for finite resources, and ignore declining fertility which occurs in all aging species. Medawar concludes that older individuals are too few in number to influence the population's gene pool for or against aging. This conclusion is found to be incorrect when variations in the age of reproductive senescence are introduced into a predator-free population.A new thought experiment with competing strains of algae corrects for these issues and shows that aging evolved and is retained so that groups retain enough genetic variability to allow for rapid evolution of a defense to novel predation. The example shows reasons why the rate of aging is directly linked to the reproductive rate, litter size, metabolic rate, reproductive senescence, and fixed body size. It also suggests that in the absence of predation, immortality would quickly evolve if not for the evolution of highly-conserved aging systems. Prior analysis of aging evolution is incorrect due to theorists' rejection of the idea of group selection. It is believed to be 'impossible' to select for mutations that are bad for the individual but good for the group. However, mutations that are neutral to young individuals which are only deleterious if expressed at older ages can accumulate in early-mortality, predator-dominated environments. Removing the predator allows deleterious mutation expression. Positive group selection then occurs amongst traits that are negative to the individual. Further, group selection is a universal force that occurs between local, non-breeding groups and not, as theorists propose, between distant groups of potentially interbreeding species. Local survivors migrate to replace extinct, related species. The antagonistic pleiotropy theory, which was created to salvage the idea of accidental aging, is examined and shown to be untenable. The hypothetical antagonistic pleiotropy genes that are beneficial to young while detrimental to old individuals, predicted to exist in the 1950s, are unlikely to exist, have not, and likely will not be found in sufficient quantity to participate in the aging process.

Aging↗
PubMed2000

Mice null for sox18 are viable and display a mild coat defect.

We have previously shown that Sox18 is expressed in developing vascular endothelium and hair follicles during mouse embryogenesis and that point mutations in Sox18 are the underlying cause of cardiovascular and hair follicle defects in ragged (Ra) mice. Here we describe the analysis of Sox18(-/-) mice produced by gene targeting. Despite the profound defects seen in Ra mice, Sox18(-/-) mice have no obvious cardiovascular defects and only a mild coat defect with a reduced proportion of zigzag hairs. A reduction in the amount of pheomelanin pigmentation in hair shafts was also observed; later-forming hair follicles showed a reduced subapical pheomelanin band, giving Sox18(-/-) mice a slightly darker appearance than Sox18(+/+) and Sox18(+/-) siblings. Sox18(-/-) mice are viable and fertile and show no difference in the ability to thrive relative to littermates. Because of the mild effect of the mutation on the phenotype of Sox18(-/-) mice, we conclude that the semidominant nature of the Ra mutations is due to a trans-dominant negative effect mediated by the mutant SOX18 proteins rather than haploinsufficiency as has been observed for other SOX genes. Due to the similarity of SOX18 to other subgroup F SOX proteins, SOX7 and -17, and the overlap in expression of these genes, functional redundancy amongst these SOX proteins could also account for the mild phenotype of Sox18(-/-) mice.

Alleles↗
PubMed2000

Frailty, family, and church support among urban African American elderly.

A community-based survey of 507 African Americans aged 60 and older from South Central Los Angeles was conducted to estimate the prevalence of frailty and describe the correlation between frailty, social support from family and church, and use of community services. Persons were considered frail if they met criteria for any of four conditions: functional impairment, depression, urinary incontinence, falls. Sixty-seven percent met criteria for frailty. Analyses revealed that frail elderly were significantly less likely to report feeling very close to family. Family contact, feeling that church was important, and receiving church support were similar for the frail and nonfrail. Frail elderly were more likely to use community services. These findings suggest that frail elderly in this population may not receive more support from family and church than nonfrail elderly. There is a need for caution when assuming families and churches in urban African American communities are able to support the most vulnerable elderly.

Activities of Daily Living↗
PubMed2000

A subtractive gene expression screen suggests a role for vanin-1 in testis development in mice.

The molecular pathways leading from indifferent mammalian gonad to either testis or ovary are not well understood. A number of genes, including the Y-linked sex determining gene SRY, have been shown to play roles in sex determination or differentiation, but there are clearly many missing elements to be found. We used suppression-subtractive hybridization to construct normalized cDNA libraries enriched for male-specific or female-specific transcripts in mouse fetal gonads. We describe the strategy used to efficiently screen these libraries for candidate sex-determination and gonadogenesis genes. One gene arising from these screens is vanin-1, which encodes a protein implicated in the induction of cell migration into the thymus. We find that vanin-1 is expressed male-specifically in Sertoli cells of the developing testis and may be involved in inducing cell migration from the adjacent mesonephros, a process known to be critical for testis development. This screening approach is likely to be applicable to the isolation and study of genes involved in a variety of developmental systems.

Amidohydrolases↗
PubMed1999

Sry requires a CAG repeat domain for male sex determination in Mus musculus.

SRY, the mammalian Y-chromosomal sex-determining gene, encodes a protein characterized by a DNA-binding and -bending domain referred to as the HMG box. Despite the pivotal role of this gene, only the HMG box region has been conserved through evolution, suggesting that SRY function depends solely on the HMG box and therefore acts as an architectural transcription factor. In mice (genus Mus) Sry also includes a large CAG trinucleotide repeat region encoding a carboxy-terminal glutamine-rich domain that acts as a transcriptional trans-activator in vitro. The absence of this or any other potential trans-activating domain in other mammals, however, has raised doubts as to its biological relevance. To test directly whether the glutamine-rich region is required for Sry function in vivo, we created truncation mutations of the Mus musculus musculus Sry gene and tested their ability to induce testis formation in XX embryos using a transgenic mouse assay. Sry constructs that encode proteins lacking the glutamine-rich region were unable to effect male sex determination, in contrast to their wild-type counterparts. We conclude that the glutamine-rich repeat domain of the mouse Sry protein has an essential role in sex determination in vivo, and that Sry may act via a fundamentally different biochemical mechanism in mice compared with other mammals.

Amino Acid Sequence↗
PubMed1999

Preoperative lymphoscintigraphy in the evaluation of squamous cell cancer of the vulva.

PURPOSE: A pilot study was undertaken to determine the lymphatic drainage of vulvar cancer using cutaneous lymphoscintigraphy. METHODS: Six patients with biopsy-proved T1 squamous cell cancer of the vulva were studied using 0.4 to 0.6 mCi Tc-99m HSA. Planar imaging was performed after patients received intradermal injections of Tc-99m HSA in a total volume of 0.4 ml at four sites around the vulvar lesion. RESULTS: Tumor locations included two midline lesions and three anterior third lesions. One tumor was located in the midthird of the labia majora. There was no clinically suspicious inguinal adenopathy in any patient. Based on classic anatomic descriptions of cutaneous lymphatic drainage, all but one patient would have been predicted to have drainage to both inguinal nodal basins. Cutaneous lymphoscintigraphy was successful in all six patients. Unilateral drainage was shown in five of six patients. Only one patient had bilateral inguinal drainage, and her tumor was located in the left anterior third of the labia minora. CONCLUSIONS: Cutaneous lymphoscintigraphy with Tc-99m HSA is easily performed and may be potentially useful in defining lymphatic basins at risk in squamous cell cancer of the vulva.

Biopsy↗
PubMed1997

SOX9 binds DNA, activates transcription, and coexpresses with type II collagen during chondrogenesis in the mouse.

Two lines of evidence suggest that the Sry-related gene Sox9 is important for chondrogenesis in mammalian embryos. Sox9 mRNA is expressed in chondrogenic condensations in mice, and mutations in human SOX9 are known to cause skeletal dysplasia. We show here that mouse SOX9 protein is able to bind to a SOX/SRY consensus motif in DNA and contains a modular transcriptional activation domain, consistent with a role for SOX9 as a transcription factor acting on genes involved in cartilage development. One such gene is Col2a1, which encodes type II collagen, the major structural component of cartilage. We have compared, in detail, the expression of Sox9 and Col2a1 during mouse development. In chondrogenic tissues the expression profiles of the two genes were remarkably similar. Coexpression was detected in some nonchondrogenic tissues such as the notochord, otic vesicle, and neural tube, but others such as heart and lung differed in their expression of the two genes. Immunohistochemistry using an antibody specific for SOX9 revealed that expression of SOX9 protein mirrored the distribution of Sox9 mRNA. Our results suggest that SOX9 protein is involved in the regulation of Col2a1 during chondrogenesis, but that this regulation is likely to depend on additional cofactors.

Animals↗
PubMed1996

Molecular genetic approaches to parasite identification: their value in diagnostic parasitology and systematics.

A wide range of approaches is available to parasitologists to aid in specific parasite identification and to formulate phylogenetic relationships. This review emphasises the usefulness of molecular genetic techniques, especially DNA-based procedures, in addressing problems of identification, characterisation and phylogeny of parasites. It should be stressed that an understanding of the various DNA approaches, techniques and target genes most likely to be effective in addressing key issues in diagnostic parasitology and systematics is still developing. Nevertheless, DNA methods clearly have great potential with regard to specificity and sensitivity, and applications will increase further with technological advance. Indeed, because of the minimal requirements for material, PCR-based methods especially should prove of immense value in future studies with parasites.

Animals↗
PubMed1996

Fresh and cryopreserved ovarian tissue samples from donors with lymphoma transmit the cancer to graft recipients.

Girls and young women who require ovariectomy or cancer therapy may consider having their own eggs, embryos or ovarian tissue stored (cryopreserved) for their own future use. Ovarian tissue is simple to collect and contains large numbers of germ cells. Transplantation of fresh and frozen-thawed ovarian tissue in healthy sheep and mice has resulted in normal live young. Similar techniques may be effective in the human but it is unclear whether ovarian tissue cryopreservation and grafting is suitable for ovaries from individuals with cancer or infections. If cancer cells were present in an ovary at the time of collection and survived cryopreservation and grafting they could establish cancer in the recipient. We therefore performed ovarian cryopreservation and transplantation trials using a mouse lymphoma model. This established that the lymphoma was transmitted by grafts of both fresh and frozen ovarian tissue. The normal healthy recipient mice died 9-43 days after receiving a small piece (1 mm3) of ovarian tissue from a donor with lymphoma. We conclude that ovarian tissue which is collected, cryopreserved and grafted while it contains cancer cells has the potential to spread the cancer to the graft recipient.

Aging↗
PubMed1995

Scintigraphic localisation of steroid injection site in plantar fasciitis.

Plantar fasciitis is a common cause of heel pain. We evaluated scintigraphic localisation of the inflammatory focus in 15 patients with this condition. Technetium-labelled bone scans precisely localised abnormal discrete areas of tracer uptake in 12 (80%) of patients in the medial and posterior aspect below the inferior surface of the calcaneum. Steroid injection at the inflammatory site abolished local tenderness and reduced pain in all 12. Our findings support an injection approach through the medial heel border posterior to the point of heel tenderness. Technetium scintigraphy may be a useful investigation to localise the steroid injection site in resistant cases of plantar fasciitis.

Fasciitis↗
PubMed1995

A molecular phylogeny of the human schistosomes.

Members of the genus Schistosoma are generally grouped on the basis of egg morphology, intermediate host specificity, and geographic origin. We have tested hypotheses based on these groupings by phylogenetic analysis of nuclear ribosomal (ITS2) and mitochondrial (COI) nucleotide sequences. Both mitochondrial and nuclear DNA data strongly support "traditional" hypotheses that (a) members of the Schistosoma haematobium group form a monophyletic clade, (b) members of the S. mansoni group form a monophyletic clade, (c) S. japonicum and S. mekongi form a monophyletic group relative to other schistosomes, and (d) the African schistosomes form a clade to the exclusion of the two Asian species.

Animals↗