Possible Role Of Hypoxia Research Articles

COVID-19 and Pulmonary Angiogenesis: The Possible Role of Hypoxia and Hyperinflammation in the Overexpression of Proteins Involved in Alveolar Vascular Dysfunction.

COVID-19 has been considered a vascular disease, and inflammation, intravascular coagulation, and consequent thrombosis may be associated with endothelial dysfunction. These changes, in addition to hypoxia, may be responsible for pathological angiogenesis. This research investigated the impact of COVID-19 on vascular function by analyzing post-mortem lung samples from 24 COVID-19 patients, 10 H1N1pdm09 patients, and 11 controls. We evaluated, through the immunohistochemistry technique, the tissue immunoexpressions of biomarkers involved in endothelial dysfunction, microthrombosis, and angiogenesis (ICAM-1, ANGPT-2, and IL-6, IL-1β, vWF, PAI-1, CTNNB-1, GJA-1, VEGF, VEGFR-1, NF-kB, TNF-α and HIF-1α), along with the histopathological presence of microthrombosis, endothelial activation, and vascular layer hypertrophy. Clinical data from patients were also observed. The results showed that COVID-19 was associated with increased immunoexpression of biomarkers involved in endothelial dysfunction, microthrombosis, and angiogenesis compared to the H1N1 and CONTROL groups. Microthrombosis and vascular layer hypertrophy were found to be more prevalent in COVID-19 patients. This study concluded that immunothrombosis and angiogenesis might play a key role in COVID-19 progression and outcome, particularly in patients who die from the disease.

The possible role of hypoxia in the affected tissue of relapsed clubfoot

Our aim was to study the expression of hypoxia-related proteins as a possible regulatory pathway in the contracted side tissue of relapsed clubfoot. We compared the expression of hypoxia-related proteins in the tissue of the contracted (medial) side of relapsed clubfoot, and in the tissue of the non-contracted (lateral) side of relapsed clubfoot. Tissue samples from ten patients were analyzed by immunohistochemistry and image analysis, Real-time PCR and Mass Spectrometry to evaluate the differences in protein composition and gene expression. We found a significant increase in the levels of smooth muscle actin, transforming growth factor-beta, hypoxia-inducible factor 1 alpha, lysyl oxidase, lysyl oxidase-like 2, tenascin C, matrix metalloproteinase-2, matrix metalloproteinase-9, fibronectin, collagen types III and VI, hemoglobin subunit alpha and hemoglobin subunit beta, and an overexpression of ACTA2, FN1, TGFB1, HIF1A and MMP2 genes in the contracted medial side tissue of clubfoot. In the affected tissue, we have identified an increase in the level of hypoxia-related proteins, together with an overexpression of corresponding genes. Our results suggest that the hypoxia-associated pathway is potentially a factor contributing to the etiology of clubfoot relapses, as it stimulates both angioproliferation and fibroproliferation, which are considered to be key factors in the progression and development of relapses.

Prolonged hypoxia induced melanotic pseudotumors in the larvae of blood-sucking mosquitoes

It was found that the presence of mosquito eggs in artificially created conditions of prolonged hypoxia causes the appearance of numerous melanotic pseudotumors in the larvae hatching from such eggs. In the cells of melanotic pseudotumors multilayer concentric membrane-like structures were found in the cytoplasm. In the immediate vicinity of such membranes, small spherical virus-like particles (VLP) with a diameter of about 30 nm were observed. The possible role of hypoxia in the development of melanotic pseudotumors of mosquito larvae is discussed

Dietary arginine modulates nonspecific immune responses in Indian Major Carp, Cirrhinus mrigala exposed to hypoxia

Hypoxia in aquatic environment is often a recurrent stressor which can affect growth and health of fishes. However, nutritional intervention can help aquatic animals to cope up with environmental stressors so as to alleviate the adverse effects. Here, we examined the impact of L-Arginine (Arg) supplementation on hypoxia-induced changes in immune responses and resistance to Aeromonas hydrophila infection in Indian Major Carp, Cirrhinus mrigala. The fish were fed with graded levels of Arg for 60 days and exposed to 72 h sublethal level of hypoxia (0.50 ± 0.16 mg/L dissolved oxygen [DO]). The six treatment groups with three replicates having different levels of Arginine such as N0.0 (0% Arg + Normoxia), H0.0 (0% Arg + Hypoxia), N0.7 (0.70% Arg + Normoxia), H0.7 (0.70% Arg + Hypoxia), N1.4 (1.40% Arg + Normoxia), and H1.4 (1.40% Arg + Hypoxia) were used in the study. Subsequently, the fish of different treatment groups were challenged with A. hydrophila. The hypoxia exposed fish exhibited significantly higher (P < .05) cumulative mortality compared to the unexposed fish, while Arg-fed groups showed reduced mortality irrespective of the level of dissolved oxygen. The albumin-globulin ratio, complement-3 response, respiratory burst, myeloperoxidase, serum antiprotease and lysozyme activities were significantly lower (P < .05) in hypoxia-exposed carp, revealing the increased susceptibility of fish to A. hydrophila is due to the immunosuppressive effect of hypoxia, which was improved in Arg-supplemented groups. We conclude that there is a possible role of hypoxia in sporadic outbreaks of bacterial diseases in carps as acute hypoxia causes immunosuppression which can be reversed by 0.7% Arg supplementation.

Hypoxia induces endothelial‑mesenchymal transition in pulmonary vascular remodeling.

It is well established that hypoxia induces epithelial-mesenchymal transition in vitro and in vivo. However, the role of hypoxia in endothelial-mesenchymal transition (EndMT), an important process in the pathogenesis of pulmonary hypertension, is not well-characterized. The present study demonstrated a significant downregulation of the endothelial marker CD31 and its co-localization with a mesenchymal marker, α-smooth muscle actin (α-SMA), in the intimal layer of small pulmonary arteries of rats exposed to chronic hypoxia. These results suggest a possible role of hypoxia in inducing EndMT in vivo. Consistent with these observations, pulmonary microvascular endothelial cells (PMVECs) cultured under hypoxic conditions exhibited a significant decrease in CD31 expression, alongside a marked increase in the expression of α-SMA and two other mesenchymal markers, collagen (Col) 1A1 and Col3A1. In addition, hypoxia promoted the proliferation and migration of α-SMA-expressing mesenchymal-like cells, but not of PMVECs. Of note, knockdown of hypoxia-inducible factor 1α (HIF-1α) effectively inhibited hypoxic induction of α-SMA, Col1A1 and the transcription factor Twist1, while rescuing hypoxic suppression of CD31; these results suggest that HIF-1α is essential for hypoxia-induced EndMT and that it serves as an upstream regulator of Twist1. Mechanistically, HIF-1α was identified to bind to the promoter of the Twist1 gene, thus activating Twist1 transcription and regulating EndMT. Collectively, the present results indicate that the HIF-1α/Twist1 pathway has a critical role in mediating the effect of hypoxia-induced EndMT in pulmonary arterial remodeling.

Hypoxia stimulates microenvironment in human embryonic stem cell through inflammatory signalling: An integrative analysis

Despite, several lines of evidence suggesting the possible role of hypoxia in stem cell development and differentiation its significance in conferring the stemness and pluripotency remains elusive. In the present study we sought to delineate the candidate genes and molecular pathways imposed during hypoxic microenvironment and its physiological relevance in tipping the balance between the niche and cellular differentiation. Integrated meta-analysis was performed between the hypoxia exposed and normal human embryonic stem cells, employing three transcriptomic cohorts (GSE35819, GSE9510 and GSE37761) retrieved from Gene expression omnibus (GEO) database. Results reveal that a total number of 12 genes were consistently differentially expressed (6up regulated and 6 down regulated) with FDR <0.05 and fold change >1.5. The Gene Ontology (GO) functions and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis was performed using DAVID. The GO analysis showed DEG significantly enriched in terms of Cellular process (GO:0009987), protein binding (GO:0005515) and cell part (GO:0044464). KEGG analysis indicated participation of genes associated with circadian rthyum regulation and PPAR signalling pathway. Further, gene-set signature (MsigDB) enrichment analysis showed positive regulation with inflammatory signals and negative association with PPAR and p53 pathway. Protein-protein network of gene modules suggests significant hub proteins viz. CTTNB1 (Degree = 18), IL8 (Degree = 15), NFKB1 (Degree = 15) and RELA (Degree = 15) in the PPI network. MCODE algorithm was used for subnetworks of the PPI network. Our integrative analysis documents the potential candidate genes which serves distinct roles influencing metabolic shift and induce inflammatory effectors contributing to hypoxic mediated stem cell niche.

Integrated cardiovascular/respiratory control in type 1 diabetes evidences functional imbalance: Possible role of hypoxia

BackgroundCardiovascular (baroreflex) and respiratory (chemoreflex) control mechanisms were studied separately in diabetes, but their reciprocal interaction (well known for diseases like heart failure) had never been comprehensively assessed. We hypothesized that prevalent autonomic neuropathy would depress both reflexes, whereas prevalent autonomic imbalance through sympathetic activation would depress the baroreflex but enhance the chemoreflexes. MethodsIn 46 type-1 diabetic subjects (7.0±0.9year duration) and 103 age-matched controls we measured the baroreflex (average of 7 methods), and the chemoreflexes, (hypercapnic: ventilation/carbon dioxide slope during hyperoxic progressive hypercapnia; hypoxic: ventilation/oxygen saturation slope during normocapnic progressive hypoxia). Autonomic dysfunction was evaluated by cardiovascular reflex tests. ResultsResting oxygen saturation and baroreflex sensitivity were reduced in the diabetic group, whereas the hypercapnic chemoreflex was significantly increased in the entire diabetic group. Despite lower oxygen saturation the hypoxic chemoreflex showed a trend toward a depression in the diabetic group. ConclusionCardio-respiratory control imbalance is a common finding in early type 1 diabetes. A reduced sensitivity to hypoxia seems a primary factor leading to reflex sympathetic activation (enhanced hypercapnic chemoreflex and baroreflex depression), hence suggesting a functional origin of cardio-respiratory control imbalance in initial diabetes.

The role of hypoxia and proton mechanism of parietal cells in the occurence of stress-induced and peptic ulcers

Based on experimental and clinical research, the authors revealed the preventive and therapeutic effect of continued intragastric delivery of molecular oxygen in case of stress-induced and peptic erosions and ulcers of stomach and duodenum. The hypothesis of possible role of hypoxia in the mechanism of injury of parietal cells is based on a consequence of transfer of protons from matrix of mitochondria and its further accumulation in cytoplasm. In this situation, the H+-dependent processes occur, particularly ATP synthesis and production of hydrochloric acid for which the protons are used.

Hypoxic mechanisms in primary headaches.

Background Hypoxia causes secondary headaches such as high-altitude headache (HAH) and headache due to acute mountain sickness. These secondary headaches mimic primary headaches such as migraine, which suggests a common link. We review and discuss the possible role of hypoxia in migraine and cluster headache. Methods This narrative review investigates the current level of knowledge on the relation of hypoxia in migraine and cluster headache based on epidemiological and experimental studies. Findings Epidemiological studies suggest that living in high-altitude areas increases the risk of migraine and especially migraine with aura. Human provocation models show that hypoxia provokes migraine with and without aura, whereas cluster headache has not been reliably induced by hypoxia. Possible pathophysiological mechanisms include hypoxia-induced release of nitric oxide and calcitonin gene-related peptide, cortical spreading depression and leakage of the blood-brain barrier. Conclusion There is a possible link between hypoxia and migraine and maybe cluster headache, but the exact mechanism is currently unknown. Provocation models of hypoxia have yielded interesting results suggesting a novel approach to study in depth the mechanism underlying hypoxia and primary headaches.

Does hypoxia play a role in infantile hemangioma?

Infantile hemangioma (IH), the most common tumor of infancy, is characterized by rapid growth during infancy, followed by spontaneous involution over 5-10years. Certain clinical observations have led to the suggestion that IH is triggered and maintained by hypoxia. We review the literature on the possible role of hypoxia in the etiology of IH, in particular, (1) the role of hypoxia inducible factor-1α (HIF-1α) and its downstream targets including GLUT-1 and VEGF; (2) the pathophysiological link between IH and retinopathy of prematurity; (3) hypoxic events in the early life including placental insufficiency, pre-eclampsia and low birthweight that have the potential to promote hypoxic stress; and (4) the evidence supporting the development of IH independent of HIF-1α. We also discuss these observations in the context of recent evidence of the crucial role of stem cells and the cytokines niche that governs their proliferation and inevitable differentiation, offering novel insights into the biology of IH. We propose that various triggers may simultaneously up-regulate HIF-1α, which is downstream of the renin-angiotensin system, specifically angiotensin II, which promotes production of HIF-1α. These developments shed light to the understanding of this enigmatic condition.

“Shaken baby syndrome” and forensic pathology

The concept of shaking baby syndrome (SBS) and the ‘‘triad’’ of encephalopathy, subdural hemorrhage (SDH), and retinal hemorrhage (RH) have become, as Dr. Byard [1] states, a hotly debated topic. SBS has come under increased scrutiny as our understanding of the mechanisms underlying the physiologic responses of neonates and infants continues to evolve. The triad can be secondary to trauma, but many forensic pathologists consider that impact is necessary to account for the signs and symptoms; as a consequence, the term SBS has been replaced by ‘‘shaken impact’’ syndrome or the more nondescript ‘‘abusive head trauma.’’ The issue is not whether or not shaking is injurious; Finnie’s experiment [2] clearly shows that smaller animals can die from repeated shaking. Instead, the issue is whether shaking alone can generate the forces associated with bridging vein rupture and SDH described in experimental models of SDH [3]. It is now well established that bridging vein rupture is not the only source of SDH in infants [4–6]. RHs also have multifactorial etiology [7, 8]. The small intradural vessels of the parasagittal dura, the falx, and tentorium are more extensive in young babies, and these vessels can leak to produce intradural and subdural hemorrhage. The extensive valveless plexus within the intracranial dura connects with the venous plexus of the spine and together form the craniospinal venous system [9]; the function of this system is not fully understood, though it is known to be involved in the maintenance of intracranial pressure, and affected by posture and respiration. This brings us to the possible role of hypoxia in SDH. While hypoxia on its own is not enough to produce SDH, there is a strong association between hypoxia and SDH [5, 6]. Additional factors such as increased venous pressure or coagulation disorders may serve to extend an intradural hemorrhage (IDH) into a thin-film SDH over the hemispheres and along the dural folds. Pathologists who do large numbers of perinatal autopsies see the close association between hypoxia and IDH/SDH [10]. Critics who sustain that the trauma of delivery has not been accounted for ignore the fact that with current obstetric care, traumatic tears of the falx or tentorium are extraordinarily rare [5]. In fact the ‘‘trauma’’ of delivery is one of hypoxia and increased intracranial pressures and not one of traumatic acceleration/deceleration. Geddes et al. [11] were the first to report that the microscopic changes seen in the brain of infants who died of inflicted head injury were hypoxic-ischemic rather than traumatic. The lack of traumatic changes in the brain made them suggest an alternative pathway for the findings of SDH and RH [12]. Since then other authors have postulated neck injury as the event that precipitates apnea and results in hypoxic changes of the brain [13]. Most of the current literature lumps together children under 2–3 years of age; however, the anatomy of the dura of infants below the age of 6 months is very different from an infant of age 2. Age-dependent changes in the cardiovascular, respiratory, immune, and coagulation systems, as well as individual variation in maturation, need to be considered when evaluating the triad in infants. We agree that ascribing intention to a combination of symptoms is absurd. While lethal craniocerebral trauma seems appropriate [1], this does not include the children This comment refers to the article available at DOI 10.1007/s12024013-9514-7.

Elevated incidence of suicide in people living at altitude, smokers and patients with chronic obstructive pulmonary disease and asthma: possible role of hypoxia causing decreased serotonin synthesis

Recent research indicates that suicide rates are elevated in those living at higher altitudes in both the United States and South Korea. A possible mechanism that was proposed is metabolic stress associated with hypoxia. This commentary discusses these results, and also the association between elevated suicide rates and other conditions associated with hypoxia (smoking, chronic obstructive pulmonary disease and asthma). Tryptophan hydroxylase may not normally be saturated with oxygen, so mild hypoxia would decrease serotonin synthesis. Low brain serotonin is known to be associated with suicide. Thus, the commentary proposes and discusses the hypothesis that decreased brain serotonin synthesis associated with hypoxia is a mechanism that may contribute to suicide in conditions causing hypoxia. Finally the commentary proposes various studies that could test aspects of this hypothesis.

Hypoxia Contributes to Melanoma Heterogeneity by Triggering HIF1α-Dependent Phenotype Switching

We have previously reported a model for melanoma progression in which oscillation between melanoma cell phenotypes characterized by invasion or proliferation is fundamental to tumor heterogeneity and disease progression. In this study we examine the possible role of hypoxia as one of the microenvironmental influences driving metastatic progression by promoting a switch from a proliferative to an invasive phenotype. Immunohistochemistry on primary human cutaneous melanoma biopsies showed intratumoral heterogeneity for cells expressing melanocytic markers, and a loss of these markers correlated with hypoxic regions. Furthermore, we show that the downregulation of melanocytic markers is dependent on hypoxia inducible factor 1α (HIF1α), a known regulator of the hypoxic response. In vitro invasion assays showed that a hypoxic environment increases the invasiveness of proliferative melanoma cell cultures in a HIF1α-dependent manner. In contrast, invasive phenotype melanoma cells showed no increase in invasive potential upon exposure to hypoxia. Thus, exposure of proliferative melanoma cells to hypoxic microenvironments is sufficient, in a HIF1α-dependent manner, to downregulate melanocytic marker expression and increase their invasive potential.

Different susceptibility of rat pancreatic alpha and beta cells to hypoxia

Insulin-producing beta cells are known to be highly susceptible to hypoxia, which is a major factor in their destruction after pancreatic islet transplantation. However, whether the glucagon-producing pancreatic islet alpha cells are sensitive to hypoxia is not known. Our objective was to compare the sensitivity of alpha and beta cells to hypoxia. Isolated rat pancreatic islets were exposed to hypoxia (1% oxygen, 94% N(2), 5% CO(2)) for 3 days. The viability of the alpha and beta cells, as well as the stimulus-specific secretion of glucagon and insulin, was evaluated. A quantitative analysis of the proportion of beta to alpha cells indicated that, under normoxic conditions, islet cells were composed mainly of beta cells (87 ± 3%) with only 13 ± 3% alpha cells. Instead, hypoxia treatment significantly increased the proportion of alpha cells (40 ± 13%) and decreased the proportion of beta cells to 60 ± 13%. Using the fluorescent TUNEL assay we found that only a few percent of beta cells and alpha cells were apoptotic in normoxia. In contrast, hypoxia induced an abundance of apoptotic beta cells (61 ± 22%) and had no effect on the level of apoptosis in alpha cells. In conclusion, this study demonstrates that hypoxia results in severe functional abnormality in both beta and alpha cells while alpha cells display significantly decreased rate of apoptosis compared to intensive apoptotic injury of beta cells. These findings have implications for the understanding of the possible role of hypoxia in the pathophysiology of diabetes.

Matrix metalloprotein-9 activation under cell-to-cell interaction between endothelial cells and monocytes: possible role of hypoxia and tumor necrosis factor-α

Matrix metalloproteinase (MMP)-9 plays an important role in cardiovascular events. However, the mechanisms underlying in vivo activation of MMP-9 are largely unknown. We investigated the secretion and activation of MMP-9 under a cell-to-cell interaction, and the effects of hypoxia and cytokine. Human umbilical vein endothelial cell (HUVEC) and THP-1 (human monocyte cell line) were cultured individually, or cocultured under normoxic and hypoxic conditions. In a coculture of HUVEC and THP-1, proMMP-9 secretion was increased twofold compared with individual culture of HUVEC and THP-1, whereas MMP-2 secretion was unchanged. The increase in proMMP-9 secretion was suppressed by antiadhesion molecule antibodies and mitogen-activated protein kinase inhibitors, PD98059 (MAPK/ERK kinase1 inhibitor) and SP600125 (Jun N-terminal kinase inhibitor). ProMMP-9 secretion was increased by tumor necrosis factor (TNF)-α at 50 ng/ml (P < 0.05) but was not activated under normoxic (20%) conditions. ProMMP-9 in coculture was activated under hypoxic (<1%) conditions, and was potentiated by TNF-α (both P < 0.05). To further investigate the mechanism of hypoxia-induced MMP-9 activation, heat shock protein (Hsp)90, which was suggested to be related to MMP-9 activation, was measured by Western blot analysis. The ratio of Hsp90 to glyceraldehyde-3-phosphate dehydrogenase was increased in hypoxic (<1%) coculture conditions with TNF-α (P < 0.05). Treatment with geldanamycin and 17-DMAG (Hsp90 inhibitor) suppressed the active form of MMP-9. Cell-to-cell interaction between endothelial cells and monocytes promotes proMMP-9 synthesis and secretion. Hypoxia and inflammation are suggested to play an important role in activating proMMP-9, presumably via Hsp90.

Abstract 4215: Hypoxia regulates the breast cancer stem/progenitor cell population

Abstract There is increasing evidence from our lab and others supporting the “cancer stem cell” hypothesis in breast cancer, which proposes that tumors are driven a small population of cells exhibiting stem cell properties. These cancer stem/progenitor cells may also be responsible for mediating tumor metastasis and resistance to cancer treatments. Recently, several new cancer therapies targeting tumor angiogenesis have been developed and validated in clinical trials. Specifically, the VEGFR and multikinase inhibitors sunitinib and sorafenib have been approved for treatment of some tumor types. However, clinical trials in breast cancer and other malignancies have demonstrated that these therapies ultimately result in drug resistance and only modestly prolong patient survival. Recent reports have suggested that these angiogenesis inhibitors may promote tumor invasiveness and metastasis in rodent breast cancer models. We hypothesize that the tumor stem/progenitor cell population increases in response to hypoxia in anti-angiogenic treated cancers, leading to increased tumor aggressiveness. We have used in vitro and in vivo assays to study the possible role of hypoxia in regulating mammary cancer stem/progenitor cells. We demonstrate that moderate hypoxia (2% oxygen) enriches for stem/progenitor cells in several breast cancer cell lines grown in vitro. The percentage of cells expressing ALDH as assessed by the Aldefluor assay approximately doubles in response to hypoxia. In vivo, sunitinib treatment increases the proportion of Aldefluor positive cells in SUM-159 and MDA-MB-231 breast cancer xenografts in NOD/SCID mice. In addition, tumor cells obtained from sunitinib treated mice exhibit enhanced tumor initiation and growth upon transplantation into secondary NOD/SCID mice. These studies indicate that tissue hypoxia results in expansion of the breast cancer stem/progenitor cell population. This may contribute to the increased aggressiveness and metastasis of cancers that evolve during therapy with angiogenesis inhibitors. If this is the case, then the addition of cancer stem cell targeting agents to antiangiogenic agents may reduce the development of treatment resistance and enhance treatment efficacy. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4215.

Effect of altitude on leptin levels, does it go up or down?

reports of a relationship between altitude and circulating levels of leptin suggest a possible role of hypoxia in leptin regulation. However, studies on the changes in plasma leptin levels at altitude are controversial, with some showing an increase and others suggesting no change or a fall in these

Confocal microscopic observations of stromal keratocytes during extended contact lens wear

Background: The aim of this study was to monitor changes in keratocyte density during extended contact lens wear and to explore the possible role of hypoxia and oedema in any changes observed. Methods: Twenty‐three neophyte myopic subjects wore a high Dk/t lens (Pure Vision) in one eye and a low Dk/t lens (Acuvue 2) in the other eye on an extended wear basis for six months. Slit‐scanning confocal microscopy and ultrasonic pachometry were performed on both eyes at baseline (before lens wear), after three and six months of lens wear, and one week after cessation of lens wear (the ‘post‐cessation’ visit). Results: No differences were established between die two lenses or between the three study visits for anterior stromal keratocyte density (KD). Posterior stromal KD was similar for the two lenses throughout the study. However, there was an overall drop in posterior KD of 14 per cent in bodl eyes at the six‐month visit, compared to die initial visit. Posterior KD at die six‐mondi visit was no different from diat at die post‐cessation visit. Corneal thickness was similar for die two lenses at die initial and post‐cessation visits, but was diree per cent greater for the eye wearing die Acuvue 2 lens at die six‐month visit. Conclusion: Extended contact lens wear causes a loss of keratocytes; diis loss cannot be attributed to contact lens‐induced hypoxia and/or oedema, or to an artifact of confocal microscopy relating to presence of residual oedema. It is postulated diat lens‐induced keratocyte loss may be related to die physical presence of contact lenses creating some mechanical effect.

Global hypoxia per se is an unusual cause of axonal injury.

Irreversible hypoxic brain damage and axonal injury are present in over 90% of fatal blunt head injuries. Given the frequency of each, difficulties arise as to whether or not they are due to different mechanisms and, as such, can be separately recognised and quantified. Recent literature has raised the possible role of hypoxia in the formation of axonal bulbs. The present study of 17 cases of cardio-respiratory arrest, 12 of status epilepticus, 3 of carbon monoxide poisoning and 12 controls was designed to test the relationship between hypoxia and axonal injury and to test the hypothesis whether or not the two entities can be separated into primary and secondary forms of traumatic brain injury. Axonal damage was seen in 9/17 and 7/12 of the cases with cardiac arrest and status epilepticus, respectively, in most of whom there was also evidence of raised intracranial pressure (ICP). All 3 cases of carbon monoxide poisoning had evidence of white matter damage in keeping with the classical pattern of selective vulnerability. It is concluded that the great majority of axonal damage identified in cases dying after cardiac arrest and status epilepticus can be attributed to raised ICP and the vascular complications of internal herniation. However, in some cases, axonal damage was seen in the absence of an elevated ICP, although its amount and distribution were different from diffuse axonal injury. In many cases there was an increase in expression of neuronal beta amyloid precursor protein.

Hypoxia upregulates VEGF production in keloid fibroblasts.

The etiology of keloid formation is diverse. They are characterized grossly as thick scar tissue that extends beyond the boundaries of the original wound. Histologically, keloids are composed of excessive collagen with an abnormally large number of partially or totally occluded microvessels. This occlusion of keloid microvessels has been hypothesized to contribute to a hypoxic microenvironment within these pathological scars. Vascular endothelial growth factor (VEGF), a potent endothelial cell mitogen, and proangiogenic cytokine have been implicated in normal and pathological wound healing. The purpose of this study was to evaluate the amount of VEGF protein production by fibroblast cell lines derived from keloids and normal human dermal skin in hypoxic compared with normoxic culture conditions. By enzyme-linked immunosorbent protein assay, VEGF was increased in both keloid and normal human dermal fibroblasts in hypoxia over normoxic controls. There was not, however, a significant difference between upregulation of VEGF protein when comparing the keloid and normal fibroblast groups. As the result of the data, alternative hypotheses for hypoxia-induced keloid formation were explored: (1) downstream modulation or signal transduction of VEGF, (2) VEGF production from cells other than fibroblasts, (3) the importance of matrix accumulation stimulated by hypoxia, or (4) increased migration of cells (other than fibroblasts) specific to keloid biology. These hypotheses may help explain the possible role of hypoxia in the pathogenesis of keloid formation. Future studies involving in situ hybridization or immunohistochemical analysis may offer greater insight into the mechanisms underlying keloid formation. Ultimately, our therapeutic goal is the utilization of biomolecular approaches for the suppression of keloid formation.