Growth hormone alone or combined with metoprolol preserves cardiac function after myocardial infarction in rats.

BACKGROUND AND OBJECTIVE: Beta-adrenoreceptor blocking agents are important for the treatment of myocardial infarction (MI). Accumulating evidence also indicates that growth hormone (GH) improves cardiac function after MI in rats. We aimed to investigate the cardiovascular effects of combined treatment in an animal model of MI. METHODS: MI was induced in rats by ligation of the left coronary artery. Three days after MI, animals were randomly assigned to one of four groups: controls (C) (n=19); GH (n=19) receiving s.c. 2 mg/kg per day rhGH; metoprolol (M) group (n=19) receiving 24 mg/kg per day and combined group (GHM) (n=20) treated with both GH (2 mg/kg per day s.c.) and M (24 mg/kg per day) for 9 days. Transthoracic echocardiography was performed before and after treatment. RESULTS: Serum levels of insulin-like growth factor I were significantly elevated in the GH-group but not in the GHM group compared to controls. Left ventricular volumes, cardiac index, systolic blood pressure, were similar in all groups. Percent changes in ejection fraction compared to baseline were; GH (6.1+/-5.0%) and GHM (6.1+/-4.2%) vs. C (-12.5+/-3.0%), P<0.01, M (-7.3+/-4.2%). The occurrence of aneurysms was not significantly different between the various treatment regimes. CONCLUSION: Treatment with growth hormone alone or in combination with metoprolol preserved left ventricular function after MI.

Decreasing ratios of phosphocreatine to beta-ATP correlates to progressive acute rejection in a concordant mouse heart to rat xenotransplantation model.

Biopsies are difficult to perform in rodent heart transplant models without compromising the graft function and therefore other means to evaluate the grafts repeatedly and noninvasively are warranted. The goal of the present study was to measure changes in ratios of high energy phosphorus containing metabolites detected with in vivo 31Phosphorous Magnetic Resonance Spectroscopy ((31)P MRS) in a xenotransplantation model and to investigate if these ratios correlated to histological signs of acute xenograft rejection. Thirty-five heart transplantations were performed (NMRI-mice to Lewis (RT1(1)) rats). Thirteen heart transplants underwent repeated daily in vivo (31)P MRS measurements and 22 grafts were measured on any of 4 postoperative days and thereafter sacrificed for histology. A modified scoring system based on Billingham's criteria was used to stage the rejection process. The median graft survival was 3.0 +/- 0.44 (median +/- SD) days (n = 17). Significant differences, both overall and interday, could be calculated for the phosphocreatine (PCr)/beta-adenosine triphosphate (beta-ATP) ratios and for the rejection score. The decreases in PCr/beta-ATP ratios correlated significantly to the progressive acute rejection process in the sacrificed grafts (P = 0.01). Further studies are indicated to establish the potential of (31)P MRS in immunosuppressed recipients of vascularized xenotransplants with prolonged graft survival.

The origin of alveolar macrophages in the transplanted lung: a longitudinal microsatellite-based study of donor and recipient DNA.

Transplanted lungs are initially populated by donor pulmonary alveolar macrophages (PAMs). These will form major antigen presenters for the recipient's suppressed immune system. They may be expected to be replaced by recipient major histocompatibility complex-compatible cells, with time. We have isolated CD14+ PAMs from bronchoalveolar lavage specimens for 6 months after transplantation and identified their origin by using microsatellite analysis. This DNA-based technology permits the reliable identification of the origin of cells from different individuals. We show that replacement of donor PAMs occurs with individual dynamics in each case. Recipient PAMs usually appeared within 2 weeks, whereas donor cells could be retained for as long as 6 months. In this limited series, there was no obvious correlation between the dynamics of this process and the occurrence of rejection episodes or infections.

Persistent high BAL fluid granulocyte activation marker levels as early indicators of bronchiolitis obliterans after lung transplant.

The major cause of mortality in the long-term in lung transplant recipients is chronic rejection. This is a fibroproliferative process in the small airways leading to obliterative bronchiolitis and progressive loss of lung function, both constituting the clinical entity bronchiolitis obliterans syndrome (BOS). Granulocyte activation has been implicated as one factor behind BOS. Granulocyte markers in bronchoalveolar lavage (BAL) fluid were prospectively and longitudinally studied in order to identify possible association with BOS. BAL fluid from 266 bronchoscopy procedures performed in twelve single lung, eight bilateral lung and five heart/lung transplant recipients were analysed. The majority (19 of 25) were studied for a period of 2 yrs after surgery. Myeloperoxidase (MPO), eosinophil cationic protein (ECP) and interleukin-8 (IL-8) levels were used as indirect markers of activation and attraction of granulocytes. Five patients developed BOS. Ninety-eight episodes of acute rejection, nine of bacterial infection, 19 of cytomegalovirus pneumonitis, nine of Pneumocystis carinii infection, two of aspergillus infection and two of respiratory syncytial virus infection were diagnosed. BOS patients had significantly higher mean levels of MPO, ECP and IL-8 compared to patients without BOS, irrespective of acute rejection status. Over time, the five patients with BOS had significantly elevated BAL fluid levels of MPO and ECP as well as neutrophil percentages, and in four patients this increase preceded the clinical diagnosis of BOS by several months. Elevated bronchoalveolar lavage fluid neutrophil percentage as well as levels of the granulocyte activation markers myeloperoxidase and eosinophil cationic protein appear to be early signs of development of BOS in lung transplant recipients.

Compromised antioxidant status and persistent oxidative stress in lung transplant recipients.

Oxidative stress may be a key feature, and hence important determinant, of tissue injury and allograft rejection in lung transplant recipients. To investigate this, we determined the antioxidant status (urate, ascorbate, thiols and alpha-tocopherol) and lipid peroxidation status (malondialdehyde) in bronchoalveolar lavage (BAL) fluid and blood serum of 19 consecutive lung transplant recipients 2 weeks and 1, 2, 3, 6, and 12 months post-surgery. BAL fluid and blood samples from 23 control subjects and blood from 8 patients two days before transplantation were obtained for comparison. Before surgery, the antioxidant status of patients was poor as serum ascorbate and total thiol concentrations were significantly (p < 0.05) lower than control subjects. Two weeks post-surgery, ascorbate and total thiol concentrations were still low and urate concentrations had fallen compared to control subjects (p < 0.01). At this time, BAL fluid urate concentration was higher (p < 0.01), ascorbate concentration was lower (p < 0.01) and reduced glutathione concentrations were similar to control subjects. MDA, a product of lipid peroxidation, was higher (p < 0.01) in both BAL fluid and serum obtained from transplant patients compared to control subjects. During the first 12 months post-surgery, little improvement in antioxidant status or extent of lipid peroxidation was seen in transplant recipients. Regression analysis indicated no difference in serum or BAL fluid antioxidant status in patients with acute rejection compared to those without. In conclusion, lung transplant recipients have a compromised antioxidant status before surgery and it remains poor for at least the first year following the operation. In addition, these patients have elevated MDA concentrations in both their lung lining fluid and blood over most of this time. Oxidative stress is not, however, a sufficiently sensitive endpoint to predict tissue rejection in this group.

Specific expression of Epstein-Barr virus in cutaneous squamous cell carcinomas from heart transplant recipients.

BACKGROUND: We investigated a Swedish group of 114 immunosuppressed cardiac allograft patients for the occurrence of posttransplant cutaneous squamous cell carcinomas. A total of 15 tumors were detected in specimens from 5 patients. METHODS: The tumors were analyzed for the presence of Epstein-Barr virus (EBV) genomes as well as EBV-specific gene expression by using three different techniques; the polymerase chain reaction (PCR), in situ hybridization, and immunohistochemistry. The material was also tested by PCR for high-risk human papilloma virus genome. RESULTS: EBV DNA could be detected by PCR in 10 of the investigated tumors, 7 of which also expressed EBV latent membrane protein 1 and/or EBV-encoded RNAs. No EBV genomes or EBV gene products could be detected in normal skin/resection margins, available from three of the tumors investigated. All tumors were negative for high-risk human papilloma virus DNA analyzed by PCR. CONCLUSIONS: In this study, we have found a high incidence of EBV-specific expression in posttransplant cutaneous squamous cell carcinomas. These results suggest that at least some of the skin cancers developing in immunocompromised heart transplant recipients are associated with EBV.

Bronchiolitis obliterans syndrome in lung transplant recipients is associated with increased neutrophil activity and decreased antioxidant status in the lung.

Long-term survival of lung transplant recipients is limited by the advent of obliterative bronchiolitis and irreversible airways obstruction, e.g. bronchiolitis obliterans syndrome (BOS). This study investigated whether inflammatory cells and their activation markers were increased in bronchoalveolar lavage (BAL) and transbronchial biopsies (TBB) from patients with BOS. Levels of antioxidants in BAL fluid were also assessed. BAL fluid and TBB from six single-lung, two bilateral-lung, and five heart-lung transplanted patients with diagnosis of BOS were compared with 13 transplant recipients without BOS. BAL fluid levels of myeloperoxidase (MPO), eosinophil cationic protein (ECP) and interleukin (IL)-8 were used as markers for the activation and attraction of neutrophils and eosinophils, respectively. Immunohistochemical staining of TBB with monoclonal antibodies to MPO and ECP (EG2) was performed. Significantly increased BAL percentages of neutrophils and levels of MPO were found in patients with BOS. The findings correlated well with the degree of monoclonal staining for MPO in TBB. BAL levels of ECP and IL-8 were significantly increased in BOS patients. BAL concentrations of the water-soluble antioxidants ascorbate, urate and glutathione were generally lower in BOS patients. The results indicate that neutrophil infiltration and activation, as well as oxidative stress, may play a role in the development and/or progression of bronchiolitis obliterans syndrome. Markers for neutrophil activation could have a potential role in monitoring disease activity in patients with this syndrome.

Inflammatory fibrosarcoma: update, reappraisal, and perspective on its place in the spectrum of inflammatory myofibroblastic tumors.

Inflammatory fibrosarcoma (commonly referred to as inflammatory myofibroblastic tumor) has become increasingly recognized as part of a spectrum of inflammatory myofibroblastic proliferations. It is a potentially locally aggressive myofibroblastic tumor that occurs predominantly in the mesentery of children and young adults. No reliable morphological parameters have been identified that predict prognosis. We evaluated the ultrastructural and immunophenotypic features of 16 cases of inflammatory fibrosarcoma and studied Ki67 (MIB1), PCNA, bcl-2, and p53 in an effort to identify prognostic markers. p53 was not detected immunohistochemically in any case. None of the markers were found to correlate with local recurrences, metastases, or tumor deaths. Low proliferative activity (Ki67 < 10%) was seen in all cases. A characteristic immunophenotype was reconfirmed in which lesional myofibroblasts stained for vimentin, alpha-smooth muscle actin, cytokeratins, and rarely desmin. Ultrastructural studies of seven cases confirmed the presence of a fibroblastic-myofibroblastic spectrum. Because inflammatory myofibroblastic tumor-inflammatory fibrosarcoma is associated with systemic symptoms, polymerase chain reaction studies for Epstein-Barr virus (EBV) and cytomegalovirus (CMV) were performed in 12 cases. Evaluable results in nine cases did not show evidence of either virus. The results of this study indicate that inflammatory fibrosarcoma has a low proliferative activity, which is in keeping with the impression that this is a low-grade sarcoma; that myofibroblasts can participate in true neoplasia; and that EBV and CMV do not play a role in the pathogenesis of inflammatory fibrosarcoma. The variable phenotype of the myofibroblast and its role in reactive and neoplastic processes are discussed. A perspective on the position of inflammatory fibrosarcoma in the spectrum of inflammatory myofibroblastic tumors is also given in light of the current study and the literature.

Inflammatory cells and activation markers in BAL during acute rejection and infection in lung transplant recipients: a prospective, longitudinal study.

Acute rejection of the transplanted lung is a clinical problem, since it decreases graft survival and predisposes the patient to chronic rejection and obliterative bronchiolitis (OB). In an earlier study, we had indications that eosinophil cationic protein (ECP) from activated eosinophils and hyaluronan (HYA) from fibroblasts were associated with acute pulmonary rejection. This prospective longitudinal study was designed to investigate whether molecules from activated inflammatory cells in bronchoalveolar lavage (BAL) fluid could serve as clinically useful diagnostic markers for acute rejection. BAL fluid from 138 bronchoscopies performed in 10 single lung, four bilateral lung and five heart-lung transplant recipients were analysed. Nine patients were studied for a period of more than 1 yr (mean 13.4 months) after surgery. Differential cell counts were made from the BAL fluid. ECP, myeloperoxidase (MPO), HYA and interleukin-8 (IL-8) were used as indirect markers for activation and attraction of eosinophils, neutrophils and fibroblasts, respectively. Fifty four episodes of acute rejection were diagnosed. Two patients developed OB. Nine episodes of bacterial infection, 13 episodes of cytomegalovirus (CMV) pneumonitis, three of Pneumocystis carinii infection and one of respiratory syncytial virus (RSV) infection were diagnosed. The mean levels of ECP, MPO, HYA and IL-8 were all higher during rejection episodes, but differences were not statistically significant compared to no rejection, when the confounding factors of time, concomitant infection, and repeated measures in the same individual had been accounted for. We could not confirm that measurements of eosinophil cationic protein, myeloperoxidase, hyaluronan and interleukin-8 in bronchoalveolar lavage fluid can be used as diagnostic markers for acute rejection in the postoperative follow-up of lung transplant recipients.

Relation between polymerase chain reaction findings and morphological changes during cytomegalovirus infection in transplanted lung.

Cytomegalovirus (CMV) can be present as a latent or productive infection resulting in disease. The polymerase chain reaction (PCR) is a sensitive technique to document the presence of CMV (DNA). Negative reactions are indicative of its absence. The presence of CMV (DNA) was assessed longitudinally in 261 transbronchial lung biopsy (TBB) specimens from 37 patients over a 6-month period. The TBB specimens from six serologically CMV-negative recipients who received lungs from serologically CMV-negative donors never showed a positive CMV-PCR(DNA) reaction during the study. Based on a study of their TBB specimens, 10 serologically CMV-positive recipients who received lungs from serologically CMV-negative donors all developed a CMV-PCR(DNA)-positive reaction and five (50%) morphologically manifested CMV disease. The remaining 21 serologically CMV-positive recipients who received lungs from serologically CMV-positive donors all developed a CMV-PCR(DNA)-positive reaction and 15 (71%) developed CMV pneumonitis. The data show that development of a positive CMV-PCR(DNA) reaction in a TBB sample within the first month after transplantation indicates a greatly increased risk of developing CMV disease. In addition, a positive CMV-PCR(DNA) reaction preceded morphologically manifest disease on average by 2 weeks. Comparisons between TBB and bronchoalveolar lavage show the former to provide a more dependable template.

Demonstration of Epstein-Barr virus DNA and human papillomavirus DNA in acetowhite lesions of the penile skin and the oral mucosa.

Oral hairy leukoplakia (OHL), thought to be caused by Epstein-Barr virus (EBV), shows similar histological and clinical features to human papillomavirus (HPV)-related acetowhite lesions of the vulva. We thus aimed to investigate the role of both HPV and EBV in men with acetowhite lesions of the penis. HPV but not EBV was significantly associated with penile acetowhite lesions showing koilocytosis compared with normal penile skin (12/20 versus 5/20, P < 0.02). HPV (5/20) and EBV (6/20) was detected in oral mucosa of some of these individuals. These results confirm an aetiological association between HPV and acetowhite penile lesions showing koilocytosis. HPV and EBV carriage in the oral mucosa is relatively common in young sexually active men.

PUVA in early mycosis fungoides may give long-term remission and delay extracutaneous spread.

We report a follow-up for 3-18 years of 24 patients with the non-infiltrated patch and infiltrated plaque stage of mycosis fungoides, treated with psoralen photochemotherapy (PUVA) and on resistant infiltration or development of tumours also with fractionated radiation therapy. All patients with patch and limited plaques showed complete remission after initial PUVA therapy. Half of the patients with patch stage mycosis fungoides and also half of the patients with plaque stage mycosis fungoides were in complete remission when the study ended. Most of them had remission periods for years after early PUVA treatment. Patients with more advanced mycosis fungoides needed repeated periods of PUVA therapy. Two patients with extensive infiltrated plaques did not reach complete remission at all during the study but progressed and finally died of their T-cell lymphoma. Another 4 patients with extensive plaque stage mycosis fungoides died after initial complete remission for a maximum of 3 years. In this investigation the clinical evaluation was made by one dermatologist and the histopathological evaluation by one pathologist. This is of importance since in the early stages of mycosis fungoides the diagnosis is challenging and may require a combination of clinical, histopathological and molecular evaluations. The identification of early disease is crucial for the rapid implementation of adequate treatment. The study shows that early PUVA therapy may delay extracutaneous spread and possibly also in some cases be curative.

Impaired endothelium-mediated vasodilatation in the peripheral vasculature of patients with acute pulmonary allograft rejection.

BACKGROUND: Experimental studies have provided evidence that, during acute pulmonary allograft rejection, endothelial dysfunction occurs not only in the transplanted lung but also in arteries of organs native to the transplant recipient. We therefore tested the hypothesis that allograft rejection leads to the release of factors into the circulation that could affect the endothelial function in lung transplant recipients. METHODS: Acetylcholine (10, 30, and 60 micrograms/min) and sodium nitroprusside (1, 3, and 6 micrograms/min) were infused into the brachial artery in nine transplant recipients (five single lung, one double lung, three heart-lung) 2 to 37 weeks after transplantation, during both acute rejection and rejection-free episodes. Changes in forearm blood flow were assessed with venous occlusion plethysmography. Plasma levels of interleukin-2, -6, and -8, endothelin-1, L-arginine, and asymmetric dimethylarginine were measured and correlated to rejection episodes. RESULTS: The vasodilatory response to acetylcholine was significantly reduced during acute rejection compared with rejection-free episodes (percentage increase from basal flow: 156% +/- 21%, 395% +/- 65%, and 585% +/- 87% during rejection versus 272% +/- 75%, 633% +/- 113%, and 933% +/- 158% during absence of rejection, p < 0.05). No statistically significant difference was found between vasodilatory responses to nitroprusside during acute rejection and rejection-free episodes. Plasma levels of L-arginine, asymmetric dimethylarginine, interleukin-6, and endothelin-1 were not significantly altered during lung rejection. CONCLUSIONS: These data indicate that a reversible peripheral decrease in endothelium-dependent vasodilatation occurs during acute rejection in lung transplant recipients. This result may be due to interactions among circulating cytokines and leukocytes activated by the rejection process and the endothelium.

Synapse-to-neuron ratio in the lateral geniculate nucleus of rats exposed chronically to ethanol.

Effects of chronic ethanol exposure on synapse-to-neuron ratio in the rat lateral geniculate nucleus were investigated. Male Sprague-Dawley rats were exposed to ethanol, using the Lieber-DeCarli liquid diets, for 4 months starting at the age of 5 weeks. Brains were perfusion-fixed, and the region containing the dorsal lateral geniculate nucleus was cut into slabs (500 microns thick) that were epoxy resin-embedded. From each rat, three slabs containing the structure were serially sectioned for electron microscopy. Using the double disector method, the study shows an unaltered synapse-to-neuron ratio in ethanol-treated rats when compared with controls. The findings are in agreement with previous studies on the visual system using the same exposure model. In contrast, a previous study has shown that the synapse-to-neuron ratio in locus ceruleus of ethanol-treated rats is reduced by 50%. Other studies have shown that, whereas the glutamatergic NMDA receptor is very sensitive to ethanol, the kainate/AMPA type of receptor is very much less so. Thus, the difference in ethanol-induced synapse elimination between the two regions may reflect this different sensitivity of the glutamatergic receptors, which are of the kainate/AMPA type in the lateral geniculate nucleus and of the NMDA type in the locus ceruleus.

Plasmapheresis as a rescue therapy to resolve cardiac rejection with vasculitis and severe heart failure. A report of five cases.

The predominant causes of late graft loss and death after cardiac transplantation are graft rejection and infection. The histopathological classification of acute rejection is based on cellular phenomena such as lymphocytic infiltration and myocyte damage. The adverse prognostic importance of vascular or humoral rejection has been reported, but there is no well-documented treatment available. In our experience, comprising 151 orthotopic transplants, five patients presented with graft rejection characterized by a lymphocytic vasculitis that did not respond to conventional therapy. Because of a deteriorating condition, in spite of vigorous antirejection treatment that included inotropic drugs and circulatory support. plasmapheresis was tried as a last, desperate means to stop the process from developing further. The clinical symptoms rapidly subsided in all five patients after the first couple of plasma exchanges. All of the patients are alive and well after 2-3.5 years of follow-up. Although the mechanism of action is unclear, plasmapheresis was beneficial in these critically ill patients.

Alterations in cortical visual evoked response following ethanol feeding in adult rats.

A study was performed on the visual-evoked response (VER) in adult rats that were given an ethanol containing liquid diet for 2 months and examined directly after the exposure period or subjected to a gradual decrease in ethanol over 3 days and total abstinence for 1 week. Control rats showed a first negative peak (N1) directly following the first positive peak (P1). In ethanol-exposed rats examined without withdrawal, the VER showed an increase in onset latency and a marked distorsion of the N1 region. The existing N1 potential was very sensitive to high-frequency stimulation. The alterations were partly normalized 1 week after withdrawal. There was no increase in latency to onset of the response or to P1. There remained an increase of latency and a reduced relative amplitude upon high-frequency stimulation of the N1 peak in ethanol-exposed rats compared with controls. The mechanisms underlying the changes in the cortical potentials are not clear, but they may be related to the cholinergic, glutamatergic/NMDA and/or noradrenergic cortical systems. The lack of persistent changes in onset and P1 latency may be related to the circumstance that the retinogeniculate impulses are transmitted over glutamatergic kainate receptors, which are relatively resistant to ethanol.

Chordal tissue in endomyocardial biopsies.

The incidence of chordal tissue in endomyocardial biopsies from heart transplant recipients and the possibility of an association between this incidence and tricuspid regurgitation or tricuspid valvular abnormalities were prospectively investigated. The biopsies were performed to detect rejection. Postoperative echocardiography and Doppler were done according to routine schedule and always when chordal tissue was found histologically, for specific evaluation of the tricuspid valve function. Chordal tissue was detected in 24 of 1,265 bites at 206 biopsies during the study period. Echocardiographically there was no significant increase in tricuspid regurgitation and no major valvular abnormality in any patient after biopsy showing chordal tissue. The presence of chordal tissue in biopsy specimens implies that the tricuspid valve is potentially at risk in biopsy. Our results, however, also showed that occasional capture of chordal tissue by the bioptome did not necessarily result in tricuspid valve dysfunction.

Decreased synapse-to-neuron ratio in rat locus ceruleus following chronic ethanol feeding.

Effects of chronic ethanol exposure on the synapse-to-neuron ratio of rat locus ceruleus were investigated. Male Sprague-Dawley rats were given an ethanol-containing liquid diet for 4 months starting at an age of 5 weeks. Littermates, given isocaloric amounts of an ethanol-free diet, served as control. The animals were perfusion fixed using a mixture of glutaraldehyde and paraformaldehyde. Synapse-to-neuron ratio was estimated by the double disector method and calculated from (Ns/A) x (Nsec -1) / (Nn/A) where Ns/A is the number of synapses per unit area estimated in a disector with a height of on section and Nsec -1 is height of the disector, i.e., the number of sections, used for estimating the number of neurons per area (Nn/A). The mean estimated synapse-to-neuron ratio was 2046 +/- 544 (SD) in ethanol-fed rats and 4291 +/- 1171 (SD) in control rats. The difference is statistically significant (p < 0.05). The finding may be of relevance for understanding the development of abuse, tolerance, drug dependence, and abstinence reactions.

Decreased axonal calibres without axonal loss in optic nerve following chronic alcohol feeding in adult rats: a morphometric study.

The effects of chronic ethanol exposure on number and calibres of optic nerve axons (and number of retinal ganglion cells) were investigated in a rat model. Male Sprague-Dawley rats were fed a liquid, ethanol-containing diet for 5, 10 and 17 weeks with littermates given isocaloric amounts of ethanol-free diet serving as controls. After fixation by perfusion, the optic nerves were imbedded in epoxy resin and sectioned for electron microscopy. Systematic random sampling was made from a cross-shaped area over the nerve. Axons within a counting frame were counted and morphometrically categorized with regard to mean diameter and the total number of axons estimated from number per area and the cross-sectional area of the nerve, which was measured using a digitizer table. According to non-parametric statistical analysis, ethanol exposure resulted in a significant reduction in mean cross-sectional area of the optic nerve and in mean axonal calibre but not in total axonal number in the ethanol-treated rats but there was no significant effect of duration of the exposure. The mean cross-sectional area of the nerve was reduced by 9%, 10% and 18% after 5, 10 and 17 weeks of exposure, respectively. The reduction in cross-sectional area appeared to be related to a proportional reduction in axonal and myelin area fractions. The findings indicate that chronic ethanol exposure results in decreased axonal calibres without axonal loss. This also implies that there is no reduction in the number of retinal ganglion cells.