Modification of the Marmarou and Foda model of diffuse axonal injury (DAI) improves percentage survival of rats at 24 h and increases the amount of DAI identified.

More than two decades ago, Marmarou published a valid model for producing diffuse axonal injury (DAI) in rats. Since then, both mild and severe injuries have been obtained by researchers using the original method and a weight of 450 g. However, the diffuse brain injuries produced in rats were only similar to those seen in humans when the rats sustained severe brain injuries. In these cases, rat mortality in the original article was around 50%, and the cause of death was prolonged apnea post-impact. Rat survival after impact is critical for studying the progression of DAI. In order to explain the cause of death in human victims with cranial trauma who do not show gross brain injury, testing for the presence of DAI is essential. Thus, in order to minimize local and cervical injuries to increase rat survival, attention should be paid to the following aspects: a wider head protector disc should be used, the head of the rat should be elevated at the time of impact, and the foam bed should be soft enough to allow the movement caused by acceleration. With our modified method, rat survival increased by 30% compared to the original model (80% versus 50%). Moreover, 85.7% of rats demonstrated DAI after 24 h of survival. With these modifications, injuries appear in the same locations as in humans; thus, the method is suitable for the study of traumatic DAI in humans.

Transrepression function of the glucocorticoid receptor regulates eyelid development and keratinocyte proliferation but is not sufficient to prevent skin chronic inflammation.

Glucocorticoids (GCs) play a key role in skin homeostasis and stress responses acting through the GC receptor (GR), which modulates gene expression by DNA binding-dependent (transactivation) and -independent (transrepression) mechanisms. To delineate which mechanisms underlie the beneficial and adverse effects mediated by GR in epidermis and other epithelia, we have generated transgenic mice that express a mutant GR (P493R, A494S), which is defective for transactivation but retains transrepression activity, under control of the keratin 5 promoter (K5-GR-TR mice). K5-GR-TR embryos exhibited eyelid opening at birth and corneal defects that resulted in corneal opacity in the adulthood. Transgenic embryos developed normal skin, although epidermal atrophy and focal alopecia was detected in adult mice. GR-mediated transrepression was sufficient to inhibit keratinocyte proliferation induced by acute and chronic phorbol 12-myristate 13-acetate exposure, as demonstrated by morphometric analyses, bromodeoxyuridine incorporation, and repression of keratin 6, a marker of hyperproliferative epidermis. These antiproliferative effects were mediated through negative interference of GR with MAPK/activator protein-1 and nuclear factor-kappaB activities, although these interactions occurred with different kinetics. However, phorbol 12-myristate 13-acetate-induced inflammation was only partially inhibited by GR-TR, which efficiently repressed IL-1beta and MMP-3 genes while weakly repressing IL-6 and TNF-alpha. Our data highlight the relevance of deciphering the mechanisms underlying GR actions on epithelial morphogenesis as well as for its therapeutic use to identify more restricted targets of GC administration.

Severe necrosis of oesophageal and gastric mucosa in fatal methanol poisoning.

Methanol is a potent neurotoxic substance that causes severe metabolic acidosis and serious neurological disorders. Most of the cases are accidental exposures to drinking beverages contaminated with methanol. There are few articles reporting pure methanol intoxication; however, it is well known that small quantities of pure methanol causes blindness and death, the minimum lethal dose being 50-100 ml.A case report is presented of a 67-year-old woman, who committed suicide by ingestion of 500 ml of absolute methanol. Despite symptomatic and supportive intensive care, the woman died 23 h after hospital admission due to metabolic acidosis and multiple organ dysfunction syndrome. A complete medico-legal autopsy was performed. Grossly, there was complete detachment of the oesophagus mucosa and brownish discolouration of the gastric mucosa. Histological findings showed diffuse haemorrhagic necrosis of the stomach mucosa and intense acute inflammatory infiltration of the lamina propria. To our knowledge, this is the first autopsy report of such severe digestive injuries. A discussion and review of the recent literature on the subject are given.

Differential distribution of cAMP-dependent protein kinase isoforms in various tissues of the bivalve mollusc Mytilus galloprovincialis.

The cAMP signalling pathway is involved in the regulation of basic physiological processes in bivalve molluscs. We had previously identified and characterized two isoforms of cAMP-dependent protein kinase (PKA) from the sea mussel Mytilus galloprovincialis that differ at their regulatory (R) subunit, namely, R(myt1) or R(myt2). Here we investigated the immunohistochemical expression of both PKA isoforms in various mussel tissues. R(myt1) and R(myt2) displayed a complementary subcellular localization. In general, R(myt1) was found to be uniformly distributed in the cytoplasm of most cell types, whereas R(myt2) appears to be localized only in the cell periphery and associated with certain cellular structures, such as the cilia of labial palps and gill filaments. Thus, both PKA isoforms appear to be non-redundant, but they have specific functions. R(myt1) was the main isoform present in catch muscle fibers, which suggests that PKA(myt1) may be the isoform involved in the regulation of the catch state. Conversely, R(myt2) was the only isoform detected in the cilia of gill filaments, indicating that PKA(myt2) could mediate the effects of cAMP on the ciliary beat frequency.

IKKß overexpression leads to pathologic lesions in stratified epithelia and exocrine glands and to tumoral transformation of oral epithelia.

Alterations in nuclear factor kappaB (NFκB) signaling have been related with several diseases and importantly also with cancer. Different animal models with increased or diminished NFκB signaling have shown that NFκB subunits and their regulators are relevant to the pathophysiology of different organs and tissues. In particular, both the deletion of the regulatory subunit ß of the kinase of the inhibitor of NFκB (IKKß) and its overexpression in epidermis lead to the development of skin inflammatory diseases not associated with tumoral lesions. In this work, we have studied the consequences of IKKß overexpression in other organs and tissues. We found that elevated IKKß levels led to altered development and functionality of exocrine glands (i.e., mammary glands) in transgenic female mice. In oral epithelia, increased IKKß expression produced lichenoid inflammation with abundant granulocytes, macrophages, and B cells, among other inflammatory cells. This inflammatory phenotype was associated with high incidence of tumoral lesions in oral epithelia, contrary to what was found in skin. Moreover, IKKß also increased the malignant progression of both spontaneous and experimentally induced oral tumors. These results highlight the importance of IKKß in epithelial and glandular homeostasis as well as in oral tumorigenesis and open the possibility that IKKß activity might be implicated in the development of oral cancer in humans.

IKKbeta leads to an inflammatory skin disease resembling interface dermatitis.

IKKbeta is a subunit of the IkappaB kinase (IKK) complex required for NF-kappaB activation in response to pro-inflammatory signals. NF-kappaB regulates the expression of many genes involved in inflammation, immunity, and apoptosis, and also controls cell proliferation and differentiation in different tissues; however, its function in skin physiopathology remains controversial. In this study we report the alterations caused by increased IKKbeta activity in skin basal cells of transgenic mice. These animals suffered chronic inflammation with abundant macrophages and other CD45(+) infiltrating cells in the skin, which resulted in epidermal basal cell injury and degeneration of hair follicles. They showed histological features characteristic of interface dermatitis (ID). This phenotype is accompanied by an increased production of inflammatory cytokines by transgenic keratinocytes. Accordingly, transcriptome studies show upregulation of genes associated with inflammatory responses. The inflammatory phenotype observed as a consequence of IKKbeta overexpression is independent of T and B lymphocytes, as it also arises in mice lacking these cell types. In summary, our data indicate the importance of IKKbeta in the development of ID and in the homeostasis of stratified epithelia. Our results also support the idea that IKKbeta might be a valid therapeutic target for the treatment of skin inflammatory diseases.