Computer modelling study of the mechanism of optic nerve injury in blunt trauma.

AIM: The potential causes of the optic nerve injury as a result of blunt object trauma, were investigated using a computer model. METHODS: A finite element model of the eye, the optic nerve, and the orbit with its content was constructed to simulate blunt object trauma. We used a model of the first phalanx of the index finger to represent the blunt body. The trauma was simulated by impacting the blunt body at the surface between the globe and the orbital wall at velocities between 2-5 m/s, and allowing it to penetrate 4-10 mm below the orbital rim. RESULTS: The impact caused rotations of the globe of up to 5000 degrees /s, lateral velocities of up to 1 m/s, and intraocular pressures (IOP) of over 300 mm Hg. The main stress concentration was observed at the insertion of the nerve into the sclera, at the side opposite to the impact. CONCLUSIONS: The results suggest that the most likely mechanisms of injury are rapid rotation and lateral translation of the globe, as well as a dramatic rise in the IOP. The strains calculated in the study should be sufficiently high to cause axonal damage and even the avulsion of the nerve. Finite element computer modelling has therefore provided important insights into a clinical scenario that cannot be replicated in human or animal experiments.

p53 and p21waf-1 expression correlates with apoptosis or cell survival in poorly differentiated, but not well-differentiated, retinoblastomas.

In human retinoblastomas, rare genetic mutations of the retinoblastoma gene cause massive cell proliferation, altered differentiation, and tumor formation; but paradoxically, this is accompanied by extensive apoptotic cell loss. We quantified the immunohistochemical distribution of p53, its downstream effector p21 (WAF-1), and apoptotic cells in 50 human retinoblastomas, within three concentric zones of sleeves of tumor cells surrounding blood vessels. In poorly differentiated retinoblastomas, both p53 expression and apoptosis increase toward the outer zone of tumor sleeves, whereas p21 expression occurs primarily within the inner zone. This staining pattern of p53 expression is reversed in well-differentiated tumors, whereas p21 staining and apoptotic cell distributions are unchanged. We detected no p53 mutations in four retinoblastomas and two retinoblastoma cell lines. We postulate that oxygen and cell "survival/growth factors" delivered via blood vessels protect retinoblastoma cells from apoptosis. In poorly differentiated tumors, apoptosis is spatially associated with increased p53 expression and may be p53 mediated, but in well-differentiated tumors, apoptosis does not colocalize with p53 and may be p53 independent. In retinoblastomas, p21 is involved not in cell death by apoptosis but in cell survival. Thus, p53 varies its expression (and by implication its function) with altered differentiation in retinoblastomas.

Regression of invasive conjunctival squamous carcinoma in an HIV-positive patient on antiretroviral therapy.

Case history of an African woman presenting with advanced HIV and a painful conjunctival lesion is presented. A conjunctival biopsy revealed invasive squamous cell carcinoma, with orbital invasion on computed tomography scan. She was commenced on antiretroviral therapy. She refused surgery to remove the eye and orbital contents (exenteration), and was referred to palliative care. Gradually, her immune status and ocular symptoms improved. At ophthalmic review, the tumour had apparently completely regressed. This unprecedented phenomenon may be due to antiretroviral therapy. Discussion covers conjunctival carcinoma and behaviour of HIV-related tumours with antiretroviral therapy. Antiretroviral drugs may offer a better alternative to disfiguring surgery in the future.

A computational study of the passive mechanisms of eye restraint during head impact trauma.

A finite element model of the eye and the orbit was used to examine the hypothesis that the orbital fat provides an important mechanism of eye stability during head trauma. The model includes the globe, the orbital fat, the extra-ocular muscles, and the optic nerve. MRI images of an adult human orbit were used to generate an idealized geometry of the orbital space. The globe was approximated as a sphere 12 mm in radius. The optic nerve and the sclera were represented as thin shells, whereas the vitreous and the orbital fat were represented as nearly incompressible solids of low stiffness. The orbital bone was modelled as a rigid shell. Frontal head impact resulting from a fall onto a hard floor was simulated by prescribing to the orbital bone a triangular acceleration pulse of 200 g (1962 m/s(2)) peak for a duration of 4.5 ms. The results show that the fat provides the crucial passive mechanism of eye restraint. The mechanism is a consequence of the fact that the fat is incompressible and that its motion is restricted by the rigidity of the orbital walls. Thus, the acceleration loads of short duration cannot generate significant distortion of the fat. In contrast, the passive muscles provide little support to the globe. When the connection between the orbital fat and the eye is absent the eye is held mainly by the optic nerve. We discuss the possible role that this loss of contact may have in some cases of the evulsion of the eye and the optic nerve.

Fusarium solani DNase is a signal for increasing expression of nonhost disease resistance response genes, hypersensitivity, and pisatin production.

The inoculation of pea endocarp tissue with the bean pathogen Fusarium solani f. sp. phaseoli results in a non-host resistance response causing a complete cessation of fungal growth within 6 to 8 h. In addition to previously reported elicitation by chitosan, we now report that components of this response are also induced by a DNase released from this fungus. A single band of protein corresponding with DNase activity elicits phytoalexin production and the accumulation of RnA homologous with the pathogenesis-related (PR) genes DRR49, DRR206, and DRR230. Both the enzyme activity and the eliciting potential of the Fusarium DNase (Fsp DNase) are heat stable but susceptible to digestion by proteinase K. Fsp DNase mimics the intact fungus in inducing resistance against F. solani f. sp. pisi. Also, Fsp DNase causes similar cytologically detectable changes in pea tissue, such as increasing hypersensitive discoloration and diminishing fluorescence of Hoechst 33342-stained nuclei and fluorescein diacetate stained cells.

Sex-steroid receptors in the diethylnitrosamine model of hepatocarcinogenesis: modifications by gonadal ablation and steroid replacement therapy.

The results of this study confirm our previous report of increased androgen receptor expression in livers of female SUAH Wistar rats during development of liver tumours induced by diethylnitrosamine (DENA). In adult female rats not treated with DENA, removal of the ovary increased liver androgen receptor levels but testosterone did not further enhance the androgen receptor status of ovariectomized rats. In normal adult males the testis and/or testosterone maintained high levels of androgen receptors but oestrogen reduced them in castrated rats. Oestrogen receptor levels were not significantly changed in either males or females by gonadectomy. Treatment of female rats with DENA for 10 and 16 weeks increased liver androgen receptors but oestrogen receptors were only reduced by 16 weeks of DENA treatment, whether the rats were intact or ovariectomized. Concentrations of liver androgen receptors were increased in intact and castrated male rats by 10 and 16 weeks of DENA treatment, an increase not seen in the previous experiments. Oestrogen appeared to inhibit both the increases in liver androgen receptor expression and liver tumour development in rats treated with the weakly carcinogenic dose of 10 weeks of DENA. However, the full carcinogenic dose of 16 weeks of DENA increased liver androgen receptors and decreased oestrogen receptors in female rats regardless of sex-steroid status. Development of malignant hepatocellular carcinoma (HCC) was associated with both an increase in liver androgen receptors and a decrease in oestrogen receptors. Maintenance of relatively high levels of liver oestrogen receptors appeared to protect the liver against development of HCC.

Effect of melanocyte stimulating hormone on human cultured choroidal melanocytes, uveal melanoma cells, and retinal epithelial cells.

PURPOSE: To establish methodology for the culture of human choroidal melanocytes to compare their responsiveness to melanocyte stimulating hormone (MSH) with that of their transformed melanoma counterparts and with that of the retinal epithelial cell. METHODS: Choroidal melanocytes from the choroid of eyes enucleated for the presence of malignant melanoma were cultured in MCDB 153 medium supplemented with insulin, transferrin, hydrocortisone, glutamine, nystatin, vitamin E, phorbol myristate acetate, bovine hypothalamic extract, cholera toxin, and chelexed fetal calf serum. RESULTS: High yields of pure spindle-shaped bipolar melanocytes were obtained with a doubling time of 3 to 4 days in nine consecutive eyes. Cells continued to divide after 4 months in culture. In contrast, uveal malignant melanoma cells grew rapidly in a relatively simple medium of Ham's F12:DMEM (1:1) supplemented with fetal calf serum, insulin, transferrin and glutamine. This medium was unable to support choroidal melanocytes. Choroidal melanocytes were DOPA-positive with appreciable tyrosinase activity that significantly increased with treatment with MSH. MSH also significantly altered the size, local density, and distribution of primary and mature melanosomes of ocular melanocytes. In contrast, uveal melanoma cells had a low level of tyrosinase activity and failed to respond to MSH with either an increase in enzyme activity or melanosome size. Retinal epithelial cells failed to show significant tyrosinase activity under the conditions studied or any increase in melanosome size in response to MSH. CONCLUSION: Ocular melanocytes show evidence of regulation by MSH and a range of mitogenic stimuli unlike the transformed melanoma cells, implying a loss of regulatory control in the latter.

Gelatinolytic metalloproteinase secretion patterns in ocular melanoma.

Fifteen posterior uveal melanoma cell lines were analyzed qualitatively for gelatinolytic and caseinolytic proteinase activity after one to five in vitro passages. All 15 cell lines secreted a gelatinolytic metalloproteinase, with an apparent molecular weight of 72 kD, into protein-free culture media; nine of these secreted an additional gelatinolytic metalloproteinase with an apparent molecular weight of 92 kD. Neither species had the ability to degrade casein. This approach may provide insight into the mechanisms of tumor metastasis in uveal melanoma.

Lysozyme distribution in human lacrimal glands and other ocular adnexa.

In an immunohistochemical study of the human lacrimal glands and other orbital adnexa, lysozyme was found to be present in the major and accessory lacrimal glands but absent from meibomian glands and conjunctival epithelium. Almost all acinar and tubular cells in major and accessory lacrimal glands contain lysozyme, although occasional cells show diminished staining for lysozyme, probably because of secretion. Only one secretory lacrimal tubuloacinar cell type is demonstrable, although two types previously have been described. Lacrimal duct cells do not contain lysozyme. The findings of this study support the concept that the tubuloacinar cells of the main and accessory lacrimal glands are the sole source of the lysozyme secreted into tears.

The ultrastructural pathological features of congenital microcoria. A case report.

We describe the light microscopic, immunohistochemical, and electron microscopic findings in a sporadic case of congenital microcoria in a 72-year-old man with senile cataract. We demonstrated a lack of myofilaments and desmin in the stromal cytoplasmic processes of the anterior pigmented cells of the iris, although other features of muscle differentiation were present in these few surviving cell processes that normally form the pupil dilator muscle. Degenerative changes in anterior pigment cells and iris stromal atrophy were thought to be late secondary features of microcoria. The findings suggest that congenital microcoria results from a defect of intermediate filaments in the terminal fetal stages of differentiation of the anterior pigmented epithelial cell of the iris, with absence of myofilaments and consequent failure of development of a functional dilator pupil muscle.

Xanthogranulomatous cholecystitis: clinicopathological study of 13 cases.

In a retrospective three year study 13 cases of xanthogranulomatous cholecystitis (XGC) (seven female, six male) were found in 724 gallbladders (1.8%), an estimated incidence of 1.7 cases per 100,000 population per annum. Symptoms often began with an episode of acute cholecystitis and persisted for up to five years. There was extension of xanthogranulomatous tissue into adjacent organs in nine cases. Three patients had fistulae from the gallbladder, one to skin, and two to the duodenum; this is the first report of this complication in XGC. In two patients XGC sufficiently resembled carcinoma for the surgeon to request intraoperative frozen section diagnosis. There was a high rate of postoperative infective complication, with one subphrenic abscess and three wound infections (one fatal), two in patients with fistulae.

ACP Best Practice No 164: Necropsy techniques in ophthalmic pathology.

Much important information can be obtained at necropsy by the pathological examination of the eye and its adnexal structures in adults and children. This information may be related to the cause of death (for example, violent shaking trauma in physical child abuse), or may pertain to disease processes affecting the eye, the orbit, and surrounding structures outside the orbit. This article reviews the technical methods used to remove the following: the vitreous (for example, for biochemistry); the eye itself (anterior approach); the eye and orbital contents (posterior approach); the eye, orbit and orbital walls; and the eye, orbital walls, and surrounding structures. The removal of the eye and adnexal ocular structures must be recognised as a "culturally sensitive" issue, which must be approached cautiously. It should only be undertaken for sound scientific reasons, with the fully informed consent of the relatives and/or the coroner (or equivalent authority), and with properly agreed procedures for the eventual retention or disposal of the ocular tissues. For this reason, this article reviews not only the scientific indications and methods for the removal of such tissues, but also the legal and ethical issues that must underpin this pathology "best practice".

Oestrogen receptor assay of cryostat sections of human breast carcinomas with simultaneous quantitative histology.

Cryostat sections of unfixed human breast carcinomas were assayed for oestrogen receptor (ER) content using an isoelectric focusing method to separate the receptor-bound oestradiol. Adjacent sections from the same tissue block were stained so that the tumour content could be estimated by point counting and the ER concentration adjusted to compensate for variations in cellularity. Elastosis was also assessed. The results confirm a positive correlation between ER values and both cellularity and elastosis. The measurement of ER in cryostat sections is relatively simple and rapid, is applicable to small tissue samples, and permits histological identification of the nature and composition of the assayed sample. The method is directly applicable to oestrogen receptor analyses of breast carcinomas in clinical laboratories with facilities for cryostat microtomy.

Splenic siderosis and parenteral iron dextran in maintenance haemodialysis patients.

The histological features of 40 spleens surgically removed from maintenance haemodialysis patients are reported. Twenty-four of the 40 (60%) showed massive iron loading and a significant direct correlation was found between iron loading and the amount of intravenous iron dextran administered. Since parenteral iron dextran appears to be a major factor in causing iron overload in haemodialysis patients its use as a method of iron replacement in these patients would appear inappropriate.

Cryostat section assay of oestrogen and progesterone receptors in meningiomas: a clinicopathological study.

Oestrogen receptors and progesterone receptors were measured in the cytosols from cryostat sections of 45 meningiomas from 40 patients (12 men, 28 women) using an isoelectric focusing technique. Near fascimile adjacent sections from the same tissue blocks were stained and examined to determine the histological subtype of the neoplasms. Appreciable levels of progesterone receptor (greater than 10 fmol/mg cytosol protein) were present in 24 (53.3%) of of the neoplasms, but no clinically important oestrogen receptor was detected in any of the tumours. Competitive binding studies on control tissue confirmed the specificity of the assay procedures. No correlation was found between progesterone receptor state and the age, sex, or menopausal state of the patients, or the histological subtype and site of the neoplasms. Four of the patients studied had multiple intracranial neoplasms, which in two were of differing progesterone receptor state. The presence of specific progesterone receptor in meningioma cytosols raises the possibility of hormonal manipulation in the treatment of this group of neoplasms.

Factors which influence necropsy requests: a psychological approach.

AIMS: To determine which factors influence a clinician's decision to request a necropsy. METHODS: Patient age, confidence in premortem diagnosis, relatives' attitudes, and conditions of necropsy practice were combined factorially (two levels each) in separate medical and surgical questionnaires based on clinical case histories. The interactions between the factors were measured by a repeated measures factorial analysis of variance for each of the two clinical groups. The influence of the clinician's interest in necropsies on these interactions was also examined by a similar method. RESULTS: Necropsies were more likely to be requested on young patients, when diagnostic confidence was low, and when relatives' attitudes were favourable. Conditions of necropsy practice did not affect the likelihood of a request and there was no apparent overall difference in necropsy requests between the two groups of clinicians. The "patient age" and "relatives" factors had less influence on the decision of the surgical group to request necropsy. This was attributed to the opportunity to "see for themselves" at operation and was supported by the finding that surgeons were very likely to request necropsies in the absence of surgical intervention. Clinicians from both groups with a high pre-existing interest in the necropsy were consistently more likely to request necropsies. CONCLUSIONS: The "case history" based questionnaires successfully measured the relative influence of multiple factors in relation to the decision of clinicians to request a necropsy. These findings suggest that any attempt to reverse the decline in necropsy rates should focus on changing the clinician's perception of the value of the modern necropsy.

Microassay for prostatic androgen receptors correlated with quantitative histological assessment.

A new microassay in which cryostat sections of prostate tissue were used to provide the source of soluble androgen receptor for biochemical assay, was devised using an isoelectric focusing method, with [3H]-mibolerone as the androgenic radioligand. Adjacent cryostat sections from the same tissue block were stained for diagnostic and quantitative histological assessment. The assay was used to illustrate variations in tissue androgen receptor concentration for correlation with epithelial cell content in benign prostate hyperplasia and prostatic cancer, and to show the effects of androgen receptor concentration of resection of prostatic tissue by electroresection. The results indicate that the heat in electroresection renders prostatic tissue unsuitable for androgen receptor assays, and suggest that knowledge of the cellular composition of carcinomatous prostates may be of importance in the full assessment of androgen receptor assay results. This method incorporates both a biochemical assay and histological assessment of the assayed tissue on near-facsimile sections, an advantage over conventional biochemical assays.

Distribution and prevalence of Mermet virus infections in the central United States.

Tests were run on 3,198 bird sera for neutralizing antibody of Mermet virus. The birds were mostly House Sparrows (Passer domesticus) captured in the central U.S. Antibody was detected in birds from Texas, Mississippi, Tennessee, Ohio, Indiana, Illinois, and Wisconsin, but not Kentucky or Missouri. Antibody prevalence differed by location and between years in similar locations. These results confirmed the widespread activity of Mermet virus in the central U.S., suggested irregular activity of the virus, and provided the first evidence that Mermet virus activity occurs in Mississippi, Indiana, and Wisconsin. No antibody to Mermet virus was found in paired sera from 966 humans with suspected arboviral infection.

Cytogenetics of iris melanomas: disparity with other uveal tract melanomas.

The chromosomal alterations of iris melanomas are poorly characterized, only one report has been detailed. Cytogenetic analysis was performed on the tumors and heparinized blood samples of three patients with iris melanomas; in one case a primary tumor and its related seedling were examined. On analysis of lymphocytes, two of the patients were found to experience a low level fragility of chromosome 9, in the region of a cutaneous melanoma susceptibility gene. All iris melanoma lesions were karyotyped. Clonal abnormalities of chromosomes 3, 5, 6, 7, 8, 9, 12, 15, 17, 18, 19, and Y were found, and in one case a large number of marker chromosomes were observed. No specific chromosomal change was common to the iris melanomas, but two cases had different abnormalities of chromosomes 5 and 18. Variations between the primary tumor and its related seedling were the acquisition of an additional chromosome 15, and a polyploid form of the cell line in the seedling. This study suggests that the most common chromosomal changes of posterior uveal melanomas are less frequent in iris melanomas. Iris melanomas also appear to experience relatively high levels of chromosomal alterations, including the formation of marker chromosomes, which is perhaps reminiscent of cutaneous melanoma.

Neurilemoma of the caruncle: a clinicopathological report.

We report a case of a neurilemoma (schwannoma) arising from the lacrimal caruncle. The clinical and light and electron microscopic features are described.