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Veterinary internships are common 1-year post-graduate clinical training programs that are offered both at veterinary colleges and in private practice settings. To promote the quality of these training programs, the American Association of Veterinary Medical Colleges (AAVMC) charged a working group to develop these internship guidelines, which were approved by the AAVMC in 2018 and have also been endorsed by the American Association of Veterinary Clinicians. These guidelines are intended to be applicable to all internships, in both academic and private practice settings, and they place particular emphasis on three aspects of internship training programs: competency-based education, intern well-being, and program outcome.

Chromosomal sex determination is widely used by vertebrates, however, only two genes have been identified as master sex-determining genes: SRY/Sry in mammals and DMY in the teleost medaka. Transfer of both genes into genetically female (XX) individuals can induce male development. However, transgenic strains have not been established in both cases because of infertility of the transgenic founders in mammals and low germline transmission rates in medaka. In this study, we used a BAC clone containing DMY in a 117 kb genomic region and two types of fluorescent marker to establish two DMY-transgenic medaka strains. In these strains, exogenous DMY is completely linked to a male phenotype and early gonadal development is not different from that of the wild-type strain. Sex-linkage analysis showed that the exogenous DMY was located on linkage group (LG) 23 in one strain and on LG 5 in the other strain, whereas the sex chromosome in medaka is on LG 1. Real-time PCR analysis indicated that these strains have multiple copies of DMY and higher DMY expression levels than the wild-type strain. These results showed that LGs 23 and 5 function as sex chromosomes in the two strains, respectively. This is not only the first example of the artificial generation of heritable sex chromosomes in vertebrates but also the first evidence showing plasticity of homomorphic sex chromosomes. This plasticity appears to be a characteristic of lower vertebrates and the underlying cause of frequent sex chromosome switching, recently reported in several fish and frog species.

INTRODUCTION The veterinary medical profession is evolving rapidly in the face of societal changes, technological advancements, economic limitations, and a greater recognition of its application in the public health sector. As emphasized during the conference An Agenda for Action: Veterinary Medicine’s Crucial Role in Biodefense and Public Health, veterinary professionals are ideally suited to address many of the headlining issues in public health, such as bioterrorist and agro-terrorist threats, emerging diseases, and food safety hazards. The critical question to be addressed is whether or not colleges and schools of veterinary medicine have programs sufficiently in place to ensure that these needs will be met by their graduates. Participants in the Summer Fellowship Program in Science, Technology, and Public Policy: Implications for Veterinary Medicine, hosted by the Virginia–Maryland Regional College of Veterinary Medicine in summer 2003, were encouraged to provide a ‘‘student perspective’’ on this issue. Eleven fellows, secondto fourth-year veterinary students from six veterinary schools, participated in the Summer Fellowship, which focused on bioterrorism, agro-terrorism, and homeland security. Students had the opportunity to meet with experts in the field of bioterrorism; veterinarians serving as fellows in the executive and legislative branches in Washington, DC; and veterinarians working for the Department of Defense, the American Veterinary Medical Association, the Association of American Veterinary Medical Colleges, industry, the US Department of Agriculture, the Food and Drug Administration, the Library of Congress, the Homeland Security Council, the Department of Health and Human Services, and the Maryland Department of Agriculture. This experience provided the fellows a unique bridge of perception and insight between their own ongoing veterinary education and these critical needs and obligations of veterinary medical education; it also serves as the basis of the ‘‘student perspective’’ described in this article.

Forty out of 41 oldsquaw carcasses collected during a 3 month avian cholera outbreak in Chesapeake Bay, USA, in 1994 were culture positive for Pasteurella multocida. Pasteurella-positive birds collected in February had greater (p ≤ 0.05) mean (geometric) liver concentrations of cadmium (7.35 versus 3.71 μg per g dry weight) and lower concentrations of selenium (9.90 versus 12.5 μg per g dry weight) than Pasteurella-positive birds collected during March and April. The mercury content of the livers and cadmium content of the kidneys did not differ (p> 0.05) between birds collected early in the die-off and those collected in March and April. The liver and kidney concentrations of metals in the Pasteurella-positive birds collected in 1994 were compared to apparently healthy oldsquaw (n = 67) collected from Chesapeake Bay during 1985--1987, because healthy oldsquaw were not collected during the avian cholera outbreak in 1994. Compared to the apparently healthy oldsquaw collected in 1985--1987, the mean concentrations of cadmium (liver 4.32 versus 2.65 μg per g dry weight and kidney 22.7 versus 11.5 μg per g dry weight) were greater (p ≤ 0.05) in the oldsquaw which succumbed to avian cholera in 1994. In contrast, the liver concentrations of selenium (11.9 versus 17.8 μg per g dry weight) and mercury (0.389 versus 1.83 μg per g dry weight) were lower (p ≤ 0.05) in the birds from the 1994 die-off than for the apparently healthy oldsquaw collected in 1985--1987. Three birds from the 1985--1987 cohort and none of the birds from the 1994 cohort had liver lead concentrations greater than 4 μg per g dry weight. The results of this study indicate a possible link between high cadmium tissue concentrations and susceptibility to avian cholera in oldsquaw

The Fulbright program was established by the US Congress to “enable the government of the United States to increase mutual understanding between the people of the United States and the people of other countries.” The Core Fulbright Scholar Program sends more than 800 US faculty and administrators to 125 countries to lecture or conduct research around the world each year. Unfortunately, only 28 faculty members from the US veterinary colleges have used Fulbright Scholar opportunities in the last 20 years (1989–2009). Considering recent worldwide events, such as the global dispersion of the Asian strain of highly pathogenic avian influenza and pandemic H1N1 2009 affecting human and animal species, the importance of awareness and education of veterinarians to such global issues is obviously urgent. Therefore, Fulbright scholarships represent an important opportunity to gain experience and bring this time-critical information back to fellow faculty and students. Veterinarians who wish to contribute to internationalization of the curricula and their campuses should consider applying for Fulbright Scholar support to launch their career in this pivotal direction. For details about the Fulbright Scholar Program, eligibility, and application procedures, please visit < http://www.cies.org/us_scholars/ >.

To calculate renal uptake of 99mTc-dimercaptosuccinic acid (DMSA) more accurately using single photon emission computed tomography (SPECT), it is necessary to estimate values of threshold level, which corresponds to the ratio of kidney to background (BG ratio). Thus the phantom and clinical studies were conducted. Six kinds of renal phantoms of 80 to 339 ml, contained different radioactivity of 37 to 485 MBq were prepared. These phantoms were placed in a larger body phantom filled with 1 to 50% of radioactivity representing background. Clinical application of this method was also performed. Results were as follows. 1) A significant correlation between phantom volumes and estimated volumes obtained using threshold level based on BG ratio calculated on tomographic images was found (r = 0.99). 2) A significant correlation between total counts in estimated volumes and radioactivities in phantoms was recognized (r = 0.94). 3) Known radioactivity in phantoms and radioactivity in estimated volumes were highly related (r = 0.98). 4) Clinical application showed valuable results in patients with renal dysfunction. Thus, this method can calculate more accurate renal uptake of 99mTc-DMSA.

Hematopoietic stem cell transplantation (SCT) is an established treatment for a variety of hematologic malignancies and some cancers. Because the survival rate in SCT recipients has significantly improved, quality of life and the late complications after SCT in long-term survivors have become increasingly important in the clinical setting.“ One of the most common late complications in SCT recipients is chronic graft-versus-host disease (cGVHD) that affects skin, lung, lacrimal and salivary glands Chronic GVHDassociated dry eye is prevalent and sometimes leads to blindness. However, clinical and histological characteristics, as well as the pathogenic process of lacrimal gland involvement in cGVHD are not well understood. The purpose of our ongoing studies is to clarify some of these issues.cGVHD, CD4+and CD8+T cells were co-localized with stromal fibroblasts in periductal area, whereas only CD8+T cells infiltrated into the acinar and ductal epithelia.

Mutations of the transforming growth factor beta-induced (TGFBI) gene whose product is called keratoepithelin (KE) have been identified in 4 major autosomal dominantly inherited corneal dystrophies. The purpose of this study was to identify the mutations in Japanese patients with these dystrophies, and to investigate the nature of corneal deposits.Mutations of the TGFBI gene were screened by polymerase chain reaction (PCR) followed by direct sequencing of the PCR products in Japanese patients clinically diagnosed as having granular corneal dystrophy, Avellino corneal dystrophy, lattice corneal dystrophy, and Reis-Bücklers' dystrophy. Corneal specimens obtained from corneal transplants were analyzed by histochemistry (Masson trichrome and Congo red stains), immunohistochemistry, and western blotting using anti KE antibody. I reviewed papers about TGFBI gene mutations previously published.The genotype/phenotype relationship of corneal dystrophies associated with mutations of the TGFBI gene is markedly evident. Avellino corneal dystrophy associated with the R 124 H mutation was the most common form of corneal stromal dystrophy in Japan. In Japan this dystrophy has been called granular corneal dystrophy up to now. Thiel-Behnke dystrophy (R 555 Q) has been also misdiagnosed as Reis-Bücklers' dystrophy. The original Reis-Bücklers' dystrophy is associated with R 124 L, which is compatible with superficial granular corneal dystrophy. Corneal deposits were associated with TGFBI products whose sizes were specific for their mutations.Mutations of the gene resulted in different types of KE aggregation accompanied with characteristic changes of processing and metabolism. The classification of these diseases according to genetic pathogenesis may be more appropriate than the use of clinical or histological findings.

To evaluate visual field loss using multifocal ERG(m-ERG), multifocal VEP(m-VEP), and Heidelberg Retina Flowmeter(HRF) in a patient with branch retinal artery occlusion(BRAO) and brain infarction.A 38-year-old man noticed inferior-nasal visual field loss in the left eye, and was referred to Keio University Hospital. He suffered from paralysis in the left leg due to brain infarction at the age of 24. However, he had not noticed visual field loss due to the brain infarction. His left fundus showed retinal edema in the area of a superior-temporal retinal artery occulusion. He was diagnosed as having BRAO. The Goldmann and Humphry perimetric examinations revealed homonymous quadrantanopia in the upper left field as well as inferior visual field defect in the left eye.Both m-ERG and m-VEP, especially second-kernel responses, were reduced in the affected retinal area of BRAO. But only m-VEP was affected in the corresponding area of homonymous quadrantanopia in the upper left field. The retinal flow in the area with BRAO evaluated by HRF was decreased in some areas and not in others, suggesting that retinal function was not necessarily consistent with retinal circulation.m-ERG and m-VEP are useful To differentiate retinal lesions from brain lesions in visual field loss.

Aquarium fish are one of the most popular pets in the United States. Understanding the use of chemotherapeutics in these animals has increased markedly in recent years, and this information is now readily accessible to the private practitioner in this article. The authors introduce the veterinary clinician to the commonly used chemotherapeutics, routes of administration, species-specific precautions, and treatment tips where appropriate.

We performed electrophysiologic tests on two patients with digitalis toxicity who first had photophobia and xanthopsia and revealed reversible reduced visual acuity and binocular central scotoma.The patients were a 72-year-old male and a 54-year-old male who had symptoms of digitalis toxicity.The corrected visual acuity was severely decreased during digitalis toxicity, 0.02 oculus dexter (OD) and 0.1 oculus sinister (OS) in case 1 and 0.04 OD and 0.2 OS in case 2. But visual acuity recovered as the blood levels of digitalis decreased to the normal level and the corrected visual acuity was 0.7 OD and 0.8 OS in case 1 and 0.8 OD and 0.9 OS in case 2. We recorded 30 Hz-flicker electroretinogram (ERG), single flash ERG, photopic ERG, and scotopic ERG when digitalis blood levels were elevated and normal. Decreased amplitudes of 30 Hz-flicker ERG and photopic ERG suggested that photoreceptor function was disturbed at digitalis toxicity and cone dysfunction was more severely disturbed than rod dysfunction.30 Hz-flicker ERG, as well as electrocardiogram and digitalis blood level, is a relatively convenient and useful measure of digitalis toxicity. It is necessary consiler toxicity when severe visual dysfunction is observed in patients with digitalis therapy.

To elucidate histopathologic features of the lacrimal gland in chronic graft-versus-host disease (GVHD) after allogeneic hematopoietic stem cell transplantation.Lacrimal gland specimens from five patients who had dry eye as part of the symptoms of chronic GVHD were examined by immunohistochemistry and transmission electron microscopy. Lacrimal gland specimens from five patients with Sjögren's syndrome (SS) were used as control samples.Lymphocytes, predominantly T cells, were found primarily in the periductal areas of the lacrimal gland from patients with chronic GVHD, whereas B cells were the dominant infiltrating cells in the acinar areas of the lacrimal gland from patients with SS. Notable findings in the lacrimal gland from patients with chronic GVHD were marked fibrosis of the glandular interstitium and an increase in the number of CD34(+) stromal fibroblasts. These findings were more prominent in patients with severe dry eye than in those with mild dry eye. Electron microscopic observations of the lacrimal gland from patients with chronic GVHD revealed that stromal fibroblasts were attached to various inflammatory cells, especially T cells, through primitive or rudimentary contacts. In addition, the presence of a well-developed rough endoplasmic reticulum in the fibroblasts and newly synthesized collagen fibrils in the extracellular matrix indicated an active production of extracellular matrix components. Electron micrographs revealed multilayered and thickened basal laminae of blood vessels, ducts, and lobules in the lacrimal gland of patients with chronic GVHD; however, these observations were infrequently observed in the lacrimal glands of patients with SS.The results suggest substantial differences in the lacrimal gland histopathology of patients with chronic GVHD and SS. In addition, it is likely that stromal fibroblasts are actively involved in the pathogenic process of chronic GVHD in the lacrimal gland by producing excessive extracellular matrix components.

Recently, the authors identified a gene, BIGH3, in which different mutations cause a group of hereditary corneal dystrophies: lattice type I and IIIA (CDLI and CDLIIIA), granular Groenouw type I (CDGGI), Avellino (CDA), and Reis-Bücklers' (CDRB). All these disorders are characterized by the progressive accumulation of corneal deposits with different structural organization. Experiments were conducted to determine the role of kerato-epithelin (KE), the product of BIGH3, in the pathogenesis of the diseases.KE-15 and KE-2, two rabbit antisera raised against peptides from the 69-364 and 426 - 682 amino acid regions of KE respectively, were used for immunohistology of the corneas obtained after keratoplasty in six CDLI patients, three CDGGI patients, and one CDA patient.The nonamyloid deposits observed in CDGGI stained intensively with KE-15 and KE-2, whereas the amyloid deposits in all analyzed CDLI corneas reacted to KE-2 but not to KE-15. In the CDA cornea, where amyloid and nonamyloid inclusions were present, positive staining with both antisera was observed.Pathologic amyloid and nonamyloid deposits observed in CDLI, CDGGI-, and CDA-affected corneas are caused by KE accumulation. Different staining patterns of amyloid and nonamyloid deposits observed with antibodies against the amino and carboxyl termini of KE suggest that two mechanisms of KE misfolding are implicated in the pathogenesis of 5q31-linked corneal dystrophies.

To evaluate the corneal phenotype of patients with corneal dystrophy and the R124H mutation in the BIGH3 gene.We examined the corneas of 24 unrelated Japanese individuals who had an R124H mutation in the BIGH3 gene. Large, discrete, granular deposits were present in the anterior stroma of all patients. They were subdivided into two types according to the appearance of the cornea. Histologic examination of the cornea after Masson trichrome and Congo red staining also was performed in specimens from patients who underwent keratoplasty.The first and the most common type of corneal findings on slit-lamp examination (20 of 24 patients) were discrete granular deposits in the anterior stromal layer and star-shaped opacities in the mid-to-deep stroma. The central subepithelial diffuse opacity increased with age. Amyloid deposits were seen mainly in the mid-to-deep stroma in five of the seven such patients evaluated. The second type of corneal appearance (four of 24 patients) was the presence of diffuse subepithelial opacities in the anterior stroma predominantly, rather than granular or linear opacities. Amyloid deposits were present in the anterior cornea of three of these four patients.The corneal lesions documented in patients with the R124H mutation were not unique to that disease but could be divided into two types, which likely represent a disease continuum. The lesion location, amount of amyloid deposition, or an interaction between the granular materials and amyloid may influence the varied appearance of the corneal lesion in patients with this disease.

To review the strategies for identifying the causative genes of inherited chorioretinal diseases.Three techniques, namely functional cloning, positional cloning, and positional candidate gene approach, have been used for this purpose. Our strategy for identifying the causative genes of inherited chorioretinal diseases is to clone retina-enriched genes, learn their chromosomal map position, and identify their expression in the retina.In the past 10 years, one gene has been cloned by functional cloning, 9 genes by positional cloning, and 19 genes by the positional candidate gene approach. Our strategy has identified 3 novel genes expressed in the retina, but they have not been associated with chorioretinal diseases.In the next century when the Human Genome Project is finished and the human genome has been sequenced completely, the positional candidate gene approach will become the predominant method of disease gene discovery. The future success of this method is predicated on increasingly dense mapping of the responsible genes by linkage analysis of multiple affected families with inherited chorioretinal diseases.

Human myocilin is identical to TIGR (trabecular meshwork inducible glucocorticoid response) which is responsible for the pathogenesis of juvenile-onset primary open angle glaucoma (GLCIA). We have isolated cDNA for mouse myocilin (Myoc) and investigated mouse myocilin gene expression in ocular tissues with in situ RNA hybridization. Hybridization signals were observed in the iris, ciliary body, trabecular meshwork, sclera, and retina in the mouse eye. The marked signals were seen in trabecular meshwork cells and the anterior portion of sclera. These findings suggest that myocilin mutation could affect the capacity of aqueous outflow and cause elevation of the intraocular pressure which is involved in the pathogenesis of glaucoma.

Recently, mutations in several genes have been identified as being responsible for the pathogenesis of autosomal recessive retinitis pigmentosa (arRP). These genes include rhodopsin, beta-subunit of rod cGMP phosphodiesterase (PDEB), alpha-subunit of rod cGMP phosphodiesterase (PDEA), and alpha-subunit of rod cGMP-gated channel. We here attempted to identify a novel mutation in the PDEB gene in Japanese arRP patients.Using the PCR-SSCP method, sequencing analysis, and restriction endonuclease digestion assay, we analyzed the PDEB gene in 17 Japanese families with non-dominant retinitis pigmentosa.A novel Ile535Asn mutation was identified in two patients in a single family and the mutation cosegregated with RP in this family. Among 90 unrelated healthy individuals, no one was identified as homozygous for this mutation, except for one individual who was found to be heterozygous.Isoleucine at codon 535 in the PDEB gene is conserved among various mammals. Missense mutations of the PDEB gene causing arRP have been reported in a limited region (codon 527-codon 699) in which codon 535 is located. Thus, the Ile535Asn mutation is an additional missense mutation which is responsible for the pathogenesis of arRP.

To determine whether the ability of transforming growth factor-beta (TGF-beta) to influence the contractile activity of corneal fibroblasts depends on their differentiation into myofibroblasts.Bovine corneal fibroblasts were cultured on collagen gel in MED 5 medium (F-12 nutrient mixture supplemented with 5% fetal bovine serum) with or without TGF-beta 1 (0.01-10 ng/ml). To evaluate the corneal fibroblast-derived contraction of collagen gel, the thickness of the gel was measured daily for 6 days. The total number of cells on the gel was counted with a Coulter counter. The detection of alpha-smooth muscle actin (alpha-SMA); a marker for myofibroblasts, on these cells was performed immunocytochemically by using a mouse monoclonal antibody against alpha-SMA. The number of myofibroblasts (alpha-SMA-positive cells) was determined.The control gels containing bovine corneal fibroblasts that were cultured with the MED 5 medium alone significantly contracted to 72.3 +/- 1.2% of their original thickness after 6 days. TGF-beta 1 increased the contraction of collagen gel mediated by bovine corneal fibroblasts in a dose-dependent manner. Approximately 0.2% of the cells on the control gels cultured with MED 5 medium alone were alpha-SMA positive. TGF-beta 1 significantly increased the expression of alpha-SMA in a dose-dependent manner. There was no significant correlation between the thickness of the collagen gel and the total number of cells. However, there was a significant negative correlation between the thickness of collagen gel and the number of myofibroblasts.TGF-beta 1 increased the contractile activity of bovine corneal fibroblasts and their ability to differentiate into myofibroblasts. Because contractile activity was correlated with differentiation, the influence of TGF-beta 1 on corneal fibroblast-induced collagen gel contraction may depend on the promotion of myofibroblast differentiation.

We report the first case of a 35-year-old Japanese man with multiple endocrine neoplasia (MEN) 2B de novo to be associated with primary open angle glaucoma.DNA was extracted from the patient's circulating leukocytes, specimens of the resected cervical lymph nodes, the neuroma of the eyelid, and of the conjunctival epithelium. Mutation was assayed by PCR/restriction enzyme and direct sequencing.The glaucoma and MEN 2B were diagnosed at the same time, when the patient was 16 years old. The glaucoma was controlled by medical treatment. The codon 918 mutation (met918thr) in exon 16 of the RET proto-oncogene associated with MEN 2B was identified in all these tissues.This patient was found to have the germline mutation at codon 918 (met918thr) in the RET proto-oncogene. The association between the RET proto-oncogene and glaucoma remains unclear, since glaucoma is a rare manifestation of MEN 2B.

reports in dogs 10 and cats 12 describe likely endocrinopathies based on physical examination and history, but without complete laboratory assessments. This report presents clinical and pathological data from a case of a steroid cell tumor in the ovary of a bat-eared fox (Otocyon megalotis) with hypercortisolism and diabetes mellitus. An 11-year-old nulliparous bat-eared fox was presented with bilateral protrusion of the nictitating membranes. The fox was obese and dehydrated and had a grade II/V systolic murmur. Abnormalities in serum biochemical parameters included elevations in blood urea nitrogen, glucose, alanine aminotransferase (ALT), alkaline phosphatase (ALP), total bilirubin, amylase, and cholesterol when compared to values obtained for this animal prior to the onset of signs (Table 1). Serum total bile acids (3.7 mmol/liter) were within reference ranges for fasting and postprandial concentrations in dogs. 1 Serum total thyroxine (2.3 mg/dl) concentrations were within the reference range for dogs. 3 Liver biopsies collected via laparotomy revealed moderate, diffuse hepatocellular hydropic degeneration suggestive of a steroid hepatopathy. The laparotomy site dehisced and became infected. The wound was managed with parenteral antibiotics, cleaning, and debridement. A low-dose dexamethasone test was performed after surgical dehiscence was nearly completely healed to evaluate the pituitary-adrenocortical axis. The baseline serum cortisol concentration was 2.8 mg/dl. Dexamethasone sodium phosphate (0.01 mg/kg) was administered intravenously. Serum cortisol concentrations were 2.4 mg/dl and 3.0 mg/dl 4 and 8 hours after dexamethasone administration, respectively. The adrenocorticotropic hormone (ACTH) concentration, assayed by radioimmunoassay, a was below detectable concentration, compared with 6.5 pmol/liter in a clinically normal conspecific. These findings were compatible with a diagnosis of a functional cortisol-secreting tumor. The blood glucose concentrations were variable, ranging from 65 to 195 mg/dl, until a marked hyperglycemia (350 mg/dl) was noted 3 months after the onset of signs. Glycosuria was also persistently present (. 100 mg/dl). Insulin

This is the report of phase III study to evaluate the safety and utility of 111In-DTPA-IgG in patients with strongly suspected inflammation/infection. One hundred and forty five patients with suspected sites of inflammation/infection were enrolled in the study. Only a few adverse experiences in one patient were reported, which were interpreted as having a possible relationship to the agent. A total of 171 suspected sites (12 in head and neck, 39 in thorax, 44 in abdomen and pelvis, 62 in musculoskeletal system, and 14 in other regions) were evaluated by investigators at each institute. Out of 171 sites, 18 were determined to be unevaluable, and 12 false negative and 5 false positive cases were observed. Overall sensitivity and specificity was 89.8% and 85.7%, respectively, and the agent proved to be effective in detecting lesions anywhere throughout the body. The analysis of data from this Phase III study indicates that 111In-DTPA-IgG is well-tolerated in patients and effective in determining focal sites of inflammation/infection although the physiological accumulation in some tissues such as the sinus or liver and possible excretion into the gastrointestinal tract may make it difficult to localize lesions.

Limbal conjunctival biopsies from 8 patients with atopic dermatitis and from 5 age-matched healthy individuals undergoing cataract or retinal detachment surgery were analyzed by light microscopy and immunological techniques. They were immuno-double labelled with anti-CD1a and anti-IgE or anti-CD23 (IgE receptor). In the specimens from atopic dermatitis 24 approximately 75% of positive anti-CD1a staining cells were double-stained by anti-IgE. Weak positive immuno-double stained cells with anti-CD23 were also observed, but less than with anti-IgE. The ratio of positive anti-IgE double-stained cells to positive anti-CD1a stained cells seemed to be parallel to serum IgE level, but not significant. The presence of IgE and CD23 (IgE receptor) on conjunctival Langerhans cells seems to have a positive effect on IgE-dependent antigen presentation.

We present a case of secondary corneal amyloidosis whose etiological mechanism was investigated by immunohistochemistry and electron microscopy. A 48-year-old woman had suffered from trichiasis in the right eye for 35 years, and developed secondary corneal amyloidosis, a phenomenon previously described but whose etiological mechanism has not been explained. Slitlamp examination of the cornea revealed a white excrescence with a diameter of 2 mm. The lesion was excised and examined by light and electron microscopy. Large deposits of an amorphous eosinophilic material were observed beneath the atrophic epithelium. Amyloid was detected in these deposits using Congo red stain, polarized light, and electron microscopy. Neither vascularization nor infiltration of inflammatory cells was observed. Immunohistochemical tests for protein AL, protein AA, prealbumin, beta 2-microglobulin and cytokeratin in paraffin sections were all negative. Characteristic findings were observed in the border zone between the basal cells and the deposits. Numerous digitiform cell processes and membrane-bound globular fragments of basal cells were seen in the superficial region of the deposits. The cell membrane of some globules was interrupted and the contents appeared to have been discharged into the stroma. These findings suggest that basal cells of the corneal epithelium provide an amyloid precursor on the stroma.

We have evaluated the effectiveness of central venous catheter placement using right atrial electrocardiography (RAECG). Consecutive patients under general anesthesia (n = 42) who required a central venous catheter underwent RAECG-guided catheter insertion procedure via right internal jugular vein. Catheter tip position was verified by post procedure portable chest radiography. Forty of 42 catheter tips were placed above the superior vena cava-right atrial junction, and none of them had its associated complications. The average insertion depth of catheters was 16.4 cm. We also attempted to predict the optimal catheter insertion depth for each patient from the previous measurements of external landmarks, but it was found to be difficult to predict reliably. In this point of view, we should use RAECG technique to make sure the proper positioning of the catheter tip.

A dual-energy (DE) quantitative computed tomography (QCT) method, the four-equation four-unknown method (DEQCT 4E-4U), was assessed and compared to single-energy (SE) QCT and standard DEQCT (two-line method). The results of this study indicate that bone mineral density (BMD) was more accurately estimated by the present method than by the SEQCT or standard DEQCT techniques on the basis of a phantom study when a large fat content was present. The results of both the phantom study and a human study also showed that the present method corrected for fat in estimating BMD in the presence of high-fat content. These findings suggest that use of this method for estimating BMD can provide useful information in studies assessing the metabolic state of bone. We propose that CT numbers estimated from excised vertebral bone marrow can serve as a soft-tissue correction for the present method.

A Phase II clinical study was performed in patients with strongly suspected focal sites of inflammation/infection to investigate clinical utility of 111In-DTPA-IgG. Neither adverse effects nor abnormal laboratory changes were noted in the all cases. Out of 59 patients, the clinical utility was evaluated in 56 patients with a total of 67 suspicious sites (19 in thorax, 13 in abdomen and pelvis, 30 in musculoskeletal system, and 5 in other regions). True positive results were obtained in 100% of lesions in the thorax (13/13) and the musculoskeletal system (26/26). There were three false negative and five false positive results. Overall sensitivity and specificity was 94.0% and 70.6%, respectively. Most of the true positive scintigram with the best image quality was acquired at Day 1 or Day 2 post-injection. A dose of 80 MBq was considered to be a practical dose for imaging. Our study indicates that 111In-DTPA-IgG is a safe and promising imaging agent for the detection of inflammation/infection, and that it is reasonable to proceed with Phase III studies to further evaluate clinical utility of the agent.

The maternal inheritance of Leber's hereditary optic neuropathy (LHON) is caused by defects in the genes of mitochondrial DNA (mtDNA). The most prevalent mtDNA mutation, present in 40% to 90% of families with this disease, is a G to A substitution at nucleotide position 11778. The rapid and accurate quantification of heteroplasmy of this mutation will help determine the relative risk for disease expression.The authors conducted screening tests for heteroplasmy in 44 visually affected patients with the 11778 mutation and 34 unaffected members of 36 Japanese families with LHON using the single-strand conformation polymorphism analysis. This method can detect even a single base difference between the sequences of wild type and mutant DNA strands. The percentage of mutant mtDNA was calculated using an image analyzer.Single-strand conformation polymorphism analysis allowed the detection of heteroplasmy ranging from 5% to 95%. Five (14%) of the 36 families showed heteroplasmy, and 14 (18%) of the 78 persons tested had heteroplasmy ranging from 10% to 94%. Seven patients with heteroplasmy with visual loss had mutant mtDNA ranging from 62% to 94%.Single-strand conformation polymorphism analysis is rapid, efficient, and accurate for detecting point mutations and quantifying heteroplasmy in mtDNA. Individuals with heteroplasmy with less than 60% of mutant mtDNA in circulating leukocytes are probably at lesser risk for developing optic atrophy.

We have identified a point mutation at nucleotide position 3460 in the ND1 gene of complex I in a Japanese pedigree with Leber's hereditary optic neuropathy by sequencing the ND genes in mitochondrial DNA. None of the 60 healthy Japanese had the 3460 mutation. The proband and his mother also had the 7444 mutation in the COI gene of complex IV and became nearly blind at age 19 with visual acuities of 0.02 OD and 0.04 OS We screened 30 patients with bilateral optic atrophy for the 3460 mutation, and identified one male patient who had the 3460 mutation in heteroplasmic fashion without carrying the 7444 mutation. He lost his sight at age 14 but it recovered to 1.2 OD and 0.7 OS about two years and half after the onset. The difference in final visual acuity between these two patients may reflect the degree of reduction in mitochondrial energy production.

AIMS--This study was designed to observe any changes to the corneal epithelium after penetrating keratoplasty. METHODS--The corneal epithelia of 26 patients were observed by specular microscopy 1 week, 1 month, 3 months, and 6 months following penetrating keratoplasty. RESULTS--After re-epithelialisation was confirmed by biomicroscopy 1 week after surgery, specular microscopy revealed many abnormal cells, including spindle shaped cells, nucleated cells, large cells, as well as irregular cell configurations. Although these abnormal findings tended to decrease with time, they were still present in some cases as much as 6 months postoperatively. Computerised morphometric analysis yielded mean cell areas of 1121 (SD 168) microns 2, 1139 (675) microns 2, 1712 (496) microns 2, and 1400 (377) microns 2 at 1 week, 1 month, 3 months, and 6 months respectively, all significantly greater than that of age matched controls (710 (151) microns 2). The shape factor decreased with time, but was still greater than the control level at 6 months. CONCLUSIONS--This study demonstrates that epithelial abnormalities persist longer than expected after penetrating keratoplasty, and that these subtle changes can be detected by specular microscopic observation, potentially allowing for modification and enhancement of the wound healing process.

Effect of insulin-like growth factor-I (IGF-I) on protein metabolism was investigated in burned rats receiving TPN. Twenty-six male SD rats were divided into two groups. IGF-I was administered to group IGF (IGF-I group, n = 14), but not to group C (Control group, n = 12). Loss of body weight after burn in group IGF was significantly lower than group C (p0.01). Cumulative nitrogen balance for 2 postburn days in group IGF was significantly higher than group C (p0.01). Rate of whole body protein turnover, synthesis and breakdown increased significantly in group IGF compared with group C. On the other hand, blood glucose was decreased significantly in group IGF (p0.05). Water balance made no significant difference between two groups. In group IGF, weight of the spleen, kidney, small intestine and colon increased significantly. Fractional synthesis rate and protein content of mucosa of the small intestine were significantly higher in group IGF than group C. From the histological point of view, mucosal layer was thickened and hyperplastic. Catabolism and surgical diabetes are caused in the surgical stress, and the administration of IGF-I are thought to be effective for improvement of those conditions. And IGF-I has the most remarkable effect on the small intestine of all organs studied in our experiment.

A method of desquamating single layers of corneal epithelial cells through the use of digitonin allowed the observation of the basal cell layer by scanning electron microscope. Basal cells of normal rabbit corneal epithelium were observed as small, round, and columnar-shaped cells, and the mean cell areas of superficial and basal cell layers were 507. 3 +/- 46.6 microns 2, and 39.5 +/- 2.8 microns 2, respectively. The mitotic rate of basal cells was also evaluated immunohistochemically using anti-PCNA (proliferating cell nuclear antigen) monoclonal antibody and the flat preparations of corneal epithelium. The mitotic rate (PCNA positive cells in the basal cell layer) was approximately 4%, and showed no difference between the center and peripheral portion of the cornea. We think these two new methods are useful for the analysis of morphological and functional characteristics of corneal epithelial basal cells.

Changes in the mitotic rate and epithelial keratin expression of corneal epithelial basal cells following corneal abrasion (7.0 mm in diameter) in rabbits were studied immunohistochemically using antiproliferating cell nuclear antigen (PCNA) monoclonal antibody and anti-epithelial keratin 1 (AE1). In the non-wounded control, the mitotic rate (PCNA positive cells in the basal cell layer) was approximately 4%, and only the superficial cells were stained by AE1 monoclonal antibody. One day after wounding, migrating epithelial cells at the leading edge, which reacted to AE1, showed low mitotic activity. At days 3 and 7, the mitotic rates of basal cells of regenerating epithelium were 3 times higher than that of controls. These basal cells displayed intensive staining with AE1, while the epithelium over the unwounded cornea exhibited a normal pattern limited to superficial cells. By 14 days after injury, the mitotic rate returned to normal and all epithelial cells expressed a normal AE1 staining pattern. Theses results suggest that regeneration of corneal epithelial basal cells involves changes in keratin expression, which might correlate with changes in the mitotic rate.

The effects of recombinant human growth hormone (rhGH) on protein metabolism were investigated. In the experimental study, 31 male Sprague-Dawley rats were divided into two groups. One group received 200 mU/day of rhGH for 3 consecutive days, before and after being burned (GH group). The other group received no rhGH as a control. Cumulative nitrogen balance after being burned was significantly higher in the GH group, and the rates of whole-body protein turnover, synthesis, and breakdown were significantly higher in the GH group. An increase of synthesis greater than that of breakdown resulted in an improved nitrogen balance in the GH group. Protein content of the liver and the gastrocnemius muscle were also significantly greater in the GH group. In the clinical study, 13 patients receiving esophagectomy for esophageal cancer were studied. Six of them received 24 U/day of rhGH for 5 consecutive postoperative days (GH group). Cumulative nitrogen balance on postoperative days was significantly higher in the GH group than in the control group. On the 3rd postoperative day, the rate of whole-body protein synthesis was significantly greater in the GH group; those of turnover and breakdown also increased in the GH group. The arteriovenous difference of amino acid composition revealed that uptake of branched-chain amino acids into the leg muscles was significantly elevated and that release of phenylalanine and tyrosine from the muscles was significantly reduced in the GH group. Hepatic function was not affected by the administration of rhGH, and rhGH inhibited the rise of blood urea nitrogen and total bilirubin after esophagectomy.(ABSTRACT TRUNCATED AT 250 WORDS)

The response of advanced prostatic cancer with metastatic chest wall tumor to high-dose diethylstilbestrol diphosphate (DESP) therapy was monitored by in vivo 31P magnetic resonance spectroscopy (31P MRS) study. A eighty-three year old man with Stage D2 prostatic cancer had been treated with chlormadinone acetate and cyclophosphamide since 1984. He was admitted to our hospital with a chest wall tumor and anemia on May 9, 1992. The elevated PAP, PSA and gamma-Sm levels were also observed. Needle biopsy of the tumor revealed poorly differentiated adenocarcinoma metastatic from the prostatic cancer. The patient received 500 mg of DESP by DIV daily for 10 days, and the tumor was reduced by 54% clinically. The abnormal PAP, PSA and gamma-Sm levels returned to almost normal range by three weeks after the initiation of high-dose DESP therapy, and regression of the tumor was confirmed by the MRI. After the first administration of DESP, the MR spectra of the chest wall tumor showed elevated peaks of phosphomonoesters and phosphodiesters. These substances are related to the membrane metabolism and their increase represents the membranous degeneration of tumor cells. The same changes continued consecuitively for three weeks, and corresponded with the regression of the tumor. In conclusion, these results suggest that in vivo 31P MRS of malignant tumors can be useful for evaluating early response to therapy prior to other clinical examinations.

A generalized biochemical deficiency of the mitochondrial matrix enzyme ornithine aminotransferase (OAT) is the inborn error in gyrate atrophy (GA), an autosomal recessive blinding disease of the retina and choroid of the eye. Because mutations in the OAT gene show a high degree of molecular heterogeneity in GA, the authors set out to determine the mutations by rapid and efficient methods.The mutations in the OAT gene were determined by a combination of polymerase chain reaction (PCR) amplification of gene sequences, analysis by denaturing gradient gel electrophoresis (DGGE), and direct DNA sequencing.Eleven different mutations in 21 (95.5%) out of 22 mutant OAT alleles from 11 patients were identified: six missense mutations, three nonsense mutations, one 2 bp-deletion, and one splice acceptor mutation. A silent polymorphism of Asn (AAC)378 to Asn (AAT) was also observed. CONCLUSIONS. The combination of PCR amplification of the gene sequences, DGGE analysis, and direct sequencing is a rapid and efficient method for detection of mutations in GA cases. The diversity of the mutations attests to the enormous genetic heterogeneity in this disease.

Two different molecular biological approaches to the disease-causing genes of genetic eye diseases are described. In gyrate atrophy of the chroid and retina where the biochemical defect was identified as inactivation of ornithine aminotransferase, the gene was cloned by using antibody for the enzyme. In most genetic eye diseases, however, the biochemical defects are unknown. Positional cloning and/or the candidate gene approach are used to identify the disease-causing genes for these diseases. The genes of chroideremia and Norrie disease were cloned by positional cloning. Several genes expressed in the photoreceptor cells have been identified recently and may be the genes causing progressive degeneration of the retina and choroid. Rhodopsin, peripherin (RDS), rom-1, and beta subunit-cGMP phosphodiesterase are identified as the disease-causing genes for retinitis pigmentosa by the candidate gene approach.