Biomedical subjects

David Greenhalgh

Publications and source records attributed to David Greenhalgh.

8 recordsLinked to original sources

PubMed2004

Alterations in the levels of metallothionein and metals in the liver, and unique serum liver enzyme response in metallothionein knock-out mice after burn injury.

OBJECTIVES: Metallothionein (MT) is a small cysteine-rich protein that sequesters and distributes metal ions. Its overexpression stimulates cell proliferation and inhibits apoptosis. We investigated the effects of burn injury on MT expression and metal localization. We also sought to determine roles of MT in the pathophysiologic alterations in the liver after injury. METHODS: Mice (C57BLKS/J, MT-I/II knock-out, KO, and wild-type control mice) were subjected to an 18% burn. Liver tissues harvested after injury were analyzed for the MT expression and the levels of zinc, copper, manganese, and iron. Levels of alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase were measured in serum samples from MT-I/MT-II KO mice and controls after injury. RESULTS: Transient induction of MT-I and MT-II mRNAs was observed 3-6 h after injury, while MT-I/MT-II protein peaked on day 1. The induction was localized to hepatocytic nuclei. The intrahepatic levels of zinc, copper, and iron were transiently elevated on day 1, when a downregulation of manganese was evident. Interestingly, only the serum levels of aspartate aminotransferase were significantly augmented in MT-I/MT-II KO mice compared to controls after injury. CONCLUSIONS: These data suggest that MT and metals may participate in the pathogenesis of the liver after burn injury.

Alanine Transaminase↗

PubMed2003

Targeted epidermal expression of mutant Connexin 26(D66H) mimics true Vohwinkel syndrome and provides a model for the pathogenesis of dominant connexin disorders.

To investigate the role of connexins in dominantly inherited skin disease, transgenic mice were produced which expressed mutant connexin 26 [gjb2/connexin 26(D66H)], from a keratin 10 promoter, exclusively in the suprabasal epidermis (the cells in which Connexin 26 is up-regulated in epidermal hyperproliferative states). From soon after birth, the mice exhibited a keratoderma similar to that in humans carrying the Connexin 26(D66H) mutation (true Vohwinkel syndrome). Transgene expression was associated with loss of Connexin 26 and Connexin 30 from epidermal keratinocyte intercellular junctions and accumulation in cytoplasm. Light and electron microscopy showed marked thickening of the epidermal cornified layers and increased epidermal TUNEL staining, indicative of premature keratinocyte programmed cell death. The K10Connexin 26(D66H) mouse may provide a valuable model to study the role of gap-junctional intercellular communication in epidermal differentiation. Similarities in phenotype between individuals (man and mouse) carrying Connexin 26(D66H) and those carrying insertional mutants of Loricrin, a major cornified envelope protein of the epidermis, suggest a possible link between connexin function and cornified envelope formation.

Animals↗

PubMed2003

Regulation of murine endogenous retroviruses in the thymus after injury.

We recently reported the induction of murine endogenous retroviruses (murine AIDS-related) in several distant organs of mice after burn injury. The regulation of endogenous retroviruses in response to burn injury was further investigated in the thymus. Female C57BLKS/J mice were subjected to 18% total body surface area flame burn injury. Thymus tissues collected at several time points (3 h to 7 days) were analyzed for the expression of subgenomic transcripts of murine endogenous retroviruses by RT-PCR. Interestingly, a novel 1.7-Kb subgenomic transcript and a recently described 1.1 Kb subgenomic transcript of murine endogenous retroviruses were transiently down-regulated in the thymus at day 1 after injury. The 1.7 Kb transcript has a coding potential for a truncated form of the envelope protein (total 214 amino acids) with a deletion of 418 amino acids near the C-terminus. The second transcript of 1.1 Kb has an open reading frame for the C-terminal transmembrane domain of the envelope protein including the p2E protein (R peptide). These data suggest the pathophysiologic effects of burn injury on the differential expression of murine endogenous retroviruses in the thymus after injury.

Amino Acid Sequence↗

PubMed2003

Injury-associated induction of two novel and replication-defective murine retroviral RNAs in the liver of mice.

Injury can alter the expression of numerous genes in affected tissues as well as in distant organs. The mouse genome harbors numerous copies of endogenous murine leukemia virus (MuLV)-related retroviral sequences. Mouse liver tissues harvested after burn injury were subjected to RT-PCR analysis to investigate the regulation of MuLV-related sequences using a primer set capable of amplifying the full-length transcript. A doublet of approximately 5-kb was transiently up-regulated at 3 and 6 h after injury. Sequence analyses revealed that these are novel defective endogenous retroviral sequences (MuLV(LI-8) and MuLV(LI-12)), which are predominantly characterized by major deletions in pol and env genes. The MuLV(LI-8) clone is 4.85 kb long and the deduced gag polypeptide sequence was almost identical to a previously reported replication-defective retroviral sequence associated with immunesuppression. In the MuLV(LI-12) clone of 5.06 kb, there were two truncated gag open reading frames (ORFs) and 1 pol ORF fused to the C-terminus of the env p15E. Furthermore, the ORFs for the unique gag p12 presumed to be responsible for the immunesuppression were present in both clones. These novel replication-defective MuLVs may participate in the pathogenesis of distant organs after injury.

Amino Acid Sequence↗

PubMed2002

The general mixing of addicts and needles in a variable-infectivity needle-sharing environment.

In this paper we develop and analyse a model for the spread of HIV/AIDS amongst a population of injecting drug users. The model we discuss focuses on the transmission of HIV through the sharing of contaminated drug injection equipment and in particular we examine the mixing of addicts and needles when the AIDS incubation period is divided into three distinct infectious stages. The impact of this assumption is to greatly increase the complexity of the HIV transmission mechanism. We begin the paper with a brief literature review followed by the derivation of a model which incorporates three classes of infectious addicts and three classes of infectious needles and where a general probability structure is used to represent the interaction of addicts and needles of varying levels of infectivity. We find that if the basic reproductive number is less than or equal to unity then there exists a globally stable disease free equilibrium. The model possesses an endemic equilibrium solution if the basic reproductive number exceeds unity. We then conduct a brief simulation study of our model. We find that the spread of disease is heavily influenced by the way addicts and needles of different levels of infectivity interact.

Computer Simulation↗

PubMed2002

Identification of truncated form of mouse HAX-1s gene (HAX-1xs) and characterization of its expression in small intestine and thymus of mice after burn injury.

Burn injury often leads to distant organ injury such as acute respiratory distress syndrome. We hypothesize that the pathophysiologic changes in distant organs result from orchestrated regulation of multiple genes in response to bum injury. Differential display was performed to identify genes regulated in distant organs in response to burn injury. Initial characterization of differentially amplified products demonstrated that HAX-1s mRNA was regulated in several distant organs after 18% total body surface area (TBSA) full-thickness flame burn injury in mice. Further characterization of HAX-1s mRNA revealed a novel transcript variant, which is rapidly and transiently induced in multiple tissues of mice within 6 h after burn injury. This novel HAX-1s transcript variant, called HAX-1xs, has an internal deletion of 252 nucleotides and single point mutation, resulting in reading frame intact. Western blot and immunohistochemical analyses of multiple tissues of mice using rabbit antibody raised against a 15-mer synthetic peptide clearly revealed the presence of HAX-1xs protein in the duodenum, and suggested that expression of HAX-1xs and/or HAX-1s was tissue- and cell type-specific. The expression of HAX-1xs and/or HAX-1s was distinctively regulated in Paneth cells of the duodenum and macrophages of the thymus after burn injury. These findings suggest that HAX-1xs has a different biological activity from HAX-1s and participates in a cascade of immediate-early cellular events in response to burn injury.

Adaptor Proteins, Signal Transducing↗

PubMedDate not supplied

The effects of a mutant connexin 26 on epidermal differentiation.

To elucidate the mode of action of dominant mutant connexins in causing inherited skin diseases, transgenic mice were produced that express the true Vohwinkel syndrome-associated mutant Cx26 (D66H), from a keratin 10 promoter, specifically in the suprabasal epidermal keratinocytes. Following birth, the transgenic mice developed keratoderma similar to that of human carriers of Cx26 (D66H). Expression of the transgene resulted in a loss of Cx26 and Cx30 at intercellular junctions of epidermal keratinocytes and accumulation of these connexins in the cytoplasm. Injection of primary mouse keratinocytes with Lucifer Yellow showed no difference in terms of dye spreading between transgenic and non transgenic keratinocytes in vitro. Expression of the mutant Cx26 (D66H) did not interfere with the formation of the epidermal water barrier during late embryonic development. Attempts to produce transgenic mice expressing the wild type form of Cx26 from the K10 promoter failed to produce viable animals although transgenic embryos were recovered at days 9 and 12 of gestation, suggesting that the transgene might be embryonic lethal.

Animals↗

PubMedDate not supplied

Prophylactic intravenous immune globulin and polymixin B decrease the incidence of septic episodes and hospital length of stay in severely burned children.

After burn shock resuscitation, serum gamma globulin levels decrease well below normal before slowly recovering over the course of 1 to 2 months. During this period, patients are vulnerable to further insult as a result of this immunocompromise. We hypothesized that intravenous immune globulin and subtherapeutic polymixin B (IVIG-B) could decrease the incidence and/or severity of sepsis after major thermal injury. A retrospective chart review from 1997 through 2003 at two hospitals compared patients who received IVIG-B (Hospital A) with those who did not (Hospital B). Patients with burns 40% or greater TBSA were included, whereas patients with nonsurvivable injuries were excluded from data analysis. A total of 152 patients were included in the study. One hundred two patients received IVIG-B, and 50 did not. Total burn size was 63.4% TBSA at Hospital A and 63.1% TBSA at Hospital B, with full-thickness burns of 54.4 and 61.7% TBSA, respectively (P < .05). Patients treated at Hospital A had a 51.9% incidence of inhalation injury compared with 28% of the patients at Hospital B (P < .05). There was an average of 1.2 and 1.9 septic episodes for patients treated at Hospital A and Hospital B, respectively (P < .05). Length of hospital stay was 77.1 days at Hospital A compared with 103.8 days at Hospital B (P < .05). Mortality was 17.6% and 18% at Hospitals A and B, respectively, and was not significantly different. Our data suggest that prophylactic IVIG-B is associated with a reduction in the incidence of septic episodes and decreased hospital length of stay following major thermal injury.

Anti-Bacterial Agents↗