Retinoblastoma epidemiology: does the evidence matter?

It has been proposed that retinoblastoma is 'caused' by two sequential mutations affecting the RB1 gene, but this is a rather outdated view of cancer aetiology that does not take into account a large amount of new acquisitions such as chromosomal and epigenetic alterations. Retinoblastoma remains probably the only cancer in which the rather simplistic 'two hit' mutational model is still considered of value, although cancer is known to be associated with genomic and microsatellite instability, defects of the DNA mismatch repair system, alterations of DNA methylation and hystone acethylation/deacethylation, and aneuploidy. Moreover, as it is shown herein, the predictions made by the 'two hit' model, are not fulfilled by the clinical and epidemiological data reported so far. Moreover, while the role of mutational events in cancer has been largely questioned in the more recent literature, no serious effort has been done to investigate the role of epigenetic alterations and aneuploidy in retinoblastoma. Through the analysis of the specialised literature and a set of original epidemiological and biological data concerning retinoblastoma, the authors illustrate the evidences arguing against the 'two hit' hypothesis and propose that epigenetic factors and aneuploidy play central roles in the disease.

Phthisis bulbi and buphthalmos as presenting signs of retinoblastoma: a report of two cases and literature review.

PURPOSE: To report two cases of bilateral retinoblastoma (RB) with unusual presentations. METHODS: The medical records of 321 patients from the Retinoblastoma Referral Center in Siena were reviewed. A total of 111 patients had bilateral RB, 2 of them presenting with phthisis bulbi and buphthalmos. Both patients underwent bilateral enucleation. Clinical features, imaging studies, and histopathology were reviewed. RESULTS: These 2 cases represent 0.62% (2/321) in our series. Histopathology did not reveal viable tumor cells in the phthisical eyes; in both buphthalmic eyes the tumor was active, infiltrating the choroid and optic nerve. CONCLUSIONS: Phthisis bulbi and buphthalmos are unusual presenting signs of RB. This very rare combination of these two signs in different eyes of the same patient is probably due to a delay in diagnosis.

Hepatic vascular response to norepinephrine during endotoxemia in anesthetized pigs.

To investigate the systemic and hepatic reactivity to norepinephrine (NE) during chronic endotoxemia, we measured the reactivity of the drug in pigs infused with endotoxin (End, 160 ng/kg/min) during an 18-h period. At the end of the experiments, the hepatic vessels were removed to test the hepatic vascular reactivity in vitro. The pressive response to NE did not change over time in control (Ctrl) pigs, but endotoxin infusion decreased the response at t = 11 and 17 h. The reactivity of the portal vein blood flow did not change in Ctrl pigs but was significantly increased in End pigs at t = 5 h. Finally, endotoxin decreased the contractile response to NE only in transversal strips of portal veins isolated from End pigs. Thus, in this model, the decreased pressive response to NE develops over time, but the modifications of the hepatic vascular reactivity remain minor.

The polymerase chain reaction (PCR) in the routine genetic characterization of retinoblastoma: a tool for the clinical laboratory.

The Polymerase Chain Reaction (PCR) is a highly innovative technique which allows for the generation of large amounts of DNA starting from minute quantities obtained from the blood or tissue of a patient. With the increasing knowledge concerning the structure of the human genome and the potential to amplify specific segments of DNA by the PCR technique, the molecular genetic characterization of many ocular disorders has been greatly facilitated. This is particularly true of retinoblastoma (RB) where the causative gene, RB1, gene has been identified and characterized. Using PCR technique, specific sequences of the RB1 gene can be amplified and analyzed to precisely define the genetic mutation in an affected individual. In addition, this technique can also be applied in order to characterize the genetic defect within the tumor itself. In this report we illustrate the use of the PCR technique in the genetic characterization of the RB1 gene and its application to the study of RB. These techniques are applicable even in a small clinical laboratory and can be extended to a number of ophthalmic disorders.

The rat isolated portal vein: a preparation sensitive to neurokinins, particularly neurokinin B.

The rat isolated portal vein is a pharmacological preparation more sensitive to neurokinin B than to any other neurokinin or tachykinin. The preparation is more sensitive to C-terminal partial sequences of substance P (SP) particularly SP-(6-11) than to the whole undecapeptide. The order of potency of neurokinins is as follows: neurokinin B greater than neurokinin A greater than substance P. The preparation shows high sensitivity also to kassinin and eledoisin. Comparative tests performed with strips of the rat portal vein suspended in a microbath under continuous perfusion (system 1) or in ordinary baths for isolated smooth muscles (system 2) have given similar results and have shown that the myotropic effect of neurokinin B is not modified by a variety of antagonists of endogenous agents as well as by inhibitors of the arachidonic acid cascade. The present results suggest that neurokinin B contracts the rat portal vein by activating specific receptors, presumably located on the smooth muscle membrane, different from those of biologically active amines and peptides which are active stimulants of the vein. Neurokinin B is ten times more active than neurokinin A and at least 100 times more than substance P. Such an order of potency of agonists suggests the existence of a new neurokinin receptor type, particularly sensitive to neurokinin B.

Pathogenesis of posterior capsular opacification. Part II: Histopathological and in vitro culture findings.

The most interesting sources of information about the pathogenesis of posterior capsular opacification seem to be histopathological studies and in vitro tissue cultures. Since our surgical technique is extracapsular cataract extraction, the explants we used for tissue culture consisted of the anterior capsule epithelial sheet without the equatorial germinative zone. We successfully overcame several problems by using the autologous plasma clot culture method. This medium, considered the optimal one for this type of culture, allowed us to study the heterogeneous behavior of the epithelial cells in culture. Using the plasma clot culture method, we were able to demonstrate in vitro fibroblastic transformation of the epithelial cells. Histopathological findings of particular cases of posterior capsule opacification and immunohistochemistry of the human lens are also reported.

Effects of clonidine and alpha-methyl-p-tyrosine on the carbachol stimulation of paradoxical sleep.

Acetylcholine promotes paradoxical sleep (PS), but the role of noradrenaline in this stimulation is controversial. The relationship between cholinergic and noradrenergic systems in the production of PS was investigated in the rat implanted on a continuous basis for sleep recordings. Stimulation of PS was obtained with microinjections of carbachol (1 microgram) into the pontine reticular formation. In the presence of the alpha 2-agonist clonidine (5 micrograms/kg, IP), the carbachol activation of PS was abolished. This stimulation also disappeared when the animals were pretreated with alpha-methyl-paratyrosine (150 mg/kg, IP), an inhibitor of catecholamine synthesis. Thus, carbachol stimulation appeared inefficient when brain noradrenergic activation was decreased. This observation supports the view that the realization of PS by the cholinergic system requires a certain level of noradrenergic activity.

Metachronous tumor development in unilateral retinoblastoma.

PURPOSE: A series of 205 retinoblastoma (RB) patients referred to the Department of Ophthalmology at the University of Siena (Italy) was evaluated in order to assess the proportion of unilateral cases later developing tumors in the companion eye ("metachronous" bilateral retinobastoma) (MBRB). METHODS: The total number of unilaterally affected patients developing tumors in the fellow eye was recorded and the risk factors assessed for the development of asynchronous bilateral retinoblastoma, i.e., family history, tumor multifocality and early age at diagnosis. RESULTS: Only two out of 133 (1.5%) unilateral retinoblastoma patients in our series could be considered affected by MBRB. CONCLUSIONS: The incidence of MBRB in our series was negligible (1.5% of all unilateral cases) compared to other reports. None of the reported risk factors for the development of tumors in the fellow eye was relevant in the present series. Although close follow-up of some unilateral cases is still recommended, thorough examination of the fellow eye, to search for lesions in the peripheral retina, is essential in all cases of unilateral RB. MBRB may be a distinctive clinical entity with specific clinical, genetic and prognostic features. However, all these aspects need to be better investigated in larger series.

Loss of heterozygosity on the long arm of chromosome 11 in orbital embryonal rhabdomyosarcoma (OERMS): a microsatellite study of seven cases.

OBJECTIVES. To investigate, by means of microsatellite analysis, regions of chromosome 11 involved in the genesis of embryonal rhabdomyosarcoma (ERMS) localized to the orbit. METHODS. Microsatellite analysis was carried out on seven cases of orbital ERMS by comparing the electrophoretic migration patterns of PCR-amplified microsatellites of chromosome 11 from both constitutional (blood) and tumor genotypes. Five of the tumors analyzed were samples frozen at the time of surgery, and two were paraffin embedded. RESULTS. Overall, microsatellites D11S1396 (11q13.1-q22.3) and D11S976 (11q) showed loss of heterozygosity (LOH) in all tumor samples, thus indicating the presence, on the long arm of chromosome 11, of one or more tumor suppressor genes with a possible role in the genesis of the disease. CONCLUSION. While the role of genes on the short arm of chromosome 11 in the genesis of ERMS is well established, much less is known of the possible involvement of tumor suppressor genes on the long arm of the same chromosome. This is the first report showing the possible involvement of tumor suppressor genes in this portion of the chromosome in ERMS localized to the orbit.

Urothelium dependent inhibition of rat ureter contractile activity.

PURPOSE: We investigated whether urothelium modulates isolated rat ureter contractions. MATERIALS AND METHODS: Segments of intact and urothelium-free ureters were placed in organ baths at 37C. The contractile effects of KCl and endogenous ureteral contractile agents were recorded in the absence and presence of the cyclooxygenase inhibitors indomethacin (1 microM) or ketoprofen (10 microM). The effect of the prostacyclin analogue iloprost was tested on the KCl and agonist induced responses obtained in the presence of ketoprofen. RESULTS: Without stimulation ureters were quiescent but spontaneous contractions often developed in urothelium-free ureters. Sensitivity to KCl was greater in the absence of urothelium. In intact ureters neurokinin A and vasopressin induced rhythmic contractions, whereas carbachol, norepinephrine, bradykinin and angiotensin II were inactive. In urothelium-free ureters the response to neurokinin A and vasopressin was enhanced and the other agonists, except norepinephrine, promoted contractions. In the presence of cyclooxygenase inhibitors intact ureters responded to carbachol, bradykinin and angiotensin II, and the response to neurokinin A, vasopressin and KCl increased. Responses obtained in urothelium-free ureters were not affected by the presence of cyclooxygenase inhibitors. In the presence of ketoprofen iloprost antagonized the KCl and agonist induced contractile effects in intact but not in urothelium-free ureters. CONCLUSIONS: Data suggest that the urothelium prevents spontaneous contractile activity and decreases the potential excitatory effects of endogenous contractile agents on ureteral motility. The mechanism underlying this inhibitory effect appears to involve the participation of a urothelial cyclooxygenase product such as prostacyclin, which could activate the release of urothelium derived relaxing factor(s) that are as yet unknown.

[Bone histomorphometry: observations in cases of renal osteodystrophy]. / Contributo allo studio dell'istomorfometria ossea: osservazioni in casi di osteodistrofia renale.

The AA report a histomorphometric study performed by measuring square areas and volumes of bone tissue. Furthermore, in 7 patients with osteodystrophy secondary to chronic renal failure, the AA investigated the amount of osteoid tissue and the presence of osteoblasts and osteoclasts. They noted signs of secondary hyperparathyroidism with increase of osteoclastic activity and fibrous metaplasia of the bone marrow associated with osteomalacia. The same patients were examined after treatment with, Vit D3 active preparations separately administered: and the AA especially noted decrease of the osteoclastic reabsorption and of the volume of osteoid tissue after the treatment with 1,25 (OH)2 D3 more than with 25 OH D3.

Everted portal vein: a sensitive model for studies of vasoactive compounds.

Three preparations of the rat hepatic portal vein, everted, noneverted, and longitudinal strip, were examined for their responsiveness to noradrenaline (NA), substance P (SP), and eledoisin (ED). The longitudinal strips and the everted veins exhibited similar sensitivities to these compounds, whereas the noneverted vein was two to four times less sensitive. The time course to generation of a maximal response was markedly slower for the noneverted vein. The poor reactivity of the noneverted vein is attributed to a reduced accessibility of the compounds to receptor sites. The myogenic responses of the longitudinal strips to NA and ED but not SP were characterized by a tonic-type contracture, whereas everted and noneverted preparations responded to the peptides, at low and intermediate concentrations, with large-amplitude and long-duration contractions. The everted vein is presented as a useful preparation for evaluation of drug-receptor interactions.

Understanding angioid streaks.

BACKGROUND: Angioid streaks are defined as a series of linear, cracked-line dehiscences of Bruch's membrane, with secondary changes in the retinal pigment epithelium and choriocapillaris. They may be progressive or degenerative, with varied presentation, color, distribution, and retinal involvement. METHODS: The epidemiology, pathophysiology, diagnosis, and management of angioid streaks are described. In addition, the systemic diseases most commonly associated with the disease are reviewed. RESULTS: Optometrists need to be able to differentially diagnose angioid streaks and to refer for evaluation of underlying systemic disease. CONCLUSIONS: Angioid streaks are considered a rare disorder, associated with pseudoxanthoma elasticum, Paget's disease of the bone, sickle hemoglobinopathies. Marfan syndrome, and Ehlers-Danlos syndrome. Ocular complications include subretinal choroidal neovascular membrane formation. Clinicians should be aware of the disease's subtle ocular appearance, its association with systemic diseases, its potential for producing subretinal ocular complications, and correct management protocols and treatments.

Postjunctional localization of substance P receptors on the rat portal vein.

A dose-dependent contractile effect of substance P (SP) on the isolated, everted rat portal vein was competitively inhibited by two selective SP antagonists (pro2, phe7, trp9)-SP and (pro4, trp7,9)-SP 4-11. Phentolamine, atropine, methysergide, mepyramine, cimetidine, Sar1, Ile8-angiotensin II, Leu8, des-Arg9-bradykinin and indomethacin did not block the action of SP. However, some of these antagonists differentially reduced SP responses, but such inhibitory effects were shown to be nonspecific. The results suggest that the SP-induced contractions of the rat portal vein were directly mediated by specific receptors localized on the smooth muscle cells. In addition, the response to SP appeared to be independent of prostaglandin biosynthesis.

[Angiographic diagnosis of mitral valve prolapse: errors and limitations]. / Diagnosi angiografica di prolasso della mitrale: errori e limiti.

In order to assess the reliability of some recently proposed methods for angiographic diagnosis and quantification of mitral valve prolapse (MVP), the left ventricular silhouettes obtained by cineangiography in 48 patients with typical auscultatory and M-mode MVP findings were examined. The following methods were tested in 40 angiographic studies that were found to be of good technical quality: method of Farry et al. (1975), of Smith et al. (1978), of Engel et al. (1978), of Spindola-Franco et al. (1979). In 23 cases a two-dimensional echocardiographic study was also performed and Engel's angiographic criteria were utilized to quantify the MVP. The angiographic and echocardiographic data were correlated in 13 cases in which it was possible to obtain both good quality echocardiographic and adequate angiographic (end-systolic identification of the two mitral valve commissures) studies. The following conclusions can be drawn: a correct angiographic quantification of the extent of bulging requires both end-diastolic and end-systolic identification of both anterolateral and posteromedial commissures; in spite of the good quality of the angiographic studies a correct end-systolic identification of the two commissures is obtainable in less than 50% of cases, which obviously restricts considerably the applicability of any objective method; among the tested angiographic methods only the Engel's method (evaluation of the distance between the plane of the mitral valve and the most protruding point of the prolapsed leaflets) showed a satisfactory diagnostic sensitivity (75% in RAO projection, 89.5% LL projection, 100% utilizing both projections); angiography is not suitable for a quantitative classification of the MVP in mild, moderate and severe forms; such a possibility, on the contrary, seems to be offered by two-dimensional echocardiography provided that Engel's angiographic are adopted.

Italian register for retinoblastoma. Pros and cons of a retrospective statistical study.

In an attempt to verify some of the current conflicting results concerning the impact of relevant prognostic factors in the retinoblastoma therapy, the authors took into consideration, for statistical analysis, the series of 459 cases included in the Italian Registry for retrospective study of retinoblastoma. Although this series appears large enough, problems related to the continuously changing approaches to the disease and the consequent lack of standardization often make it difficult to draw significant conclusions. Hence, while historical (retrospective) analysis often allows the manipulation of a great number of data, particularly in the case of relatively rare diseases, prospective randomized controlled trials are strongly recommended to standardize definitely the relevant prognostic criteria. These and other problems related to retrospective analysis are discussed in detail.

Electrophysiological properties of intravenous metoprolol in man.

Electrophysiological changes produced by intravenous (0.1 mg/kg) metoprolol, a new selective beta 1-blocking agent devoid of intrinsic activity, were studied in 16 subjects with estimated normal impulse formation and conduction. The most important effects were sinus bradycardia, mild increase of sinoatrial conduction time, depression of intranodal conduction, and prolongation of AV node refractory periods. Sinus node recovery time and atrial refractory periods were unmodified. Infranodal conduction and the refractory periods of the His-Purkinje system, as well as of the bundle-branches, were unchanged. These effects are compared with those observed after intravenous propranolol, pindolol, and oxprenolol.

Sepsis decreases the spontaneous and agonist-induced contractile activities in the rat portal vein.

BACKGROUND/AIMS: The portal vein has spontaneous and agonist-induced contractile activities and whether sepsis alters these two types of contractile activities is unknown. METHODS: To study the effect of sepsis on the spontaneous contractile activity and the contractile responses to norepinephrine (NE), angiotensin II (AT(II)), and neurokinin B (NKB) in the rat portal vein (RPV), we performed a cecal ligature and puncture (CLP) 24 h before RPV isolation. RESULTS: CLP decreased the spontaneous activity and induced hyporesponsiveness to AT(II) and NKB. The vascular failure was correlated to the severity of sepsis. In contrast, the reactivity to NE was not altered. Although inducible NO synthase was detected in RPV isolated from CLP rats, NO synthase inhibitors did not restore either the responsiveness to AT(II) and NKB or the spontaneous activity. Additionally, hyporesponsiveness to AT(II) and NKB was not modified by indomethacin. CONCLUSIONS: CLP decreases the spontaneous activity of the RPV as well as the contractile responses to AT(II) and NKB. The vascular failure is correlated to the severity of sepsis. The reactivity to NE is not altered in this model. Neither NO nor prostaglandins are responsible for the vascular abnormalities observed during CLP.

Diclofenac and NS-398, a selective cyclooxygenase-2 inhibitor, decrease agonist-induced contractions of the pig isolated ureter.

Non-steroidal anti-inflammatory drugs (NSAIDs) are currently considered a first-line treatment of renal colic. Their action has been ascribed to the inhibition of renal prostaglandin synthesis, which decreases renal blood flow and diuresis, and consequently lowers the pressure in the renal pelvis and ureter. However, the effects of NSAIDs on induced contractions of ureteral smooth muscle have received little attention. Also, there is a lack of clinically relevant spasmolytic drugs for the ureter. Therefore, we studied the influence of the non-selective cyclooxygenase (COX) inhibitor diclofenac, a NSAID drug customarily used in the treatment of renal colic, and of NS-398, a selective COX-2 inhibitor, on induced contractions of the pig ureter. Serotonin (0.1-30 microM), norepinephrine (0.1-30 microM) and neurokinin A (0.03-10 microM) induced reproducible concentration-dependent contractions, which were inhibited by diclofenac and NS-398 (10-300 microM) in a concentration-dependent manner. The sensitivity of neurokinin A-induced contractions to diclofenac was 3-4 times greater than that of the amines. Depending on the concentration, inhibition ranged between 25 and 96% of the initially induced contractile activity. In the presence of inhibitors, supramaximal concentrations of agonists were unable to trigger recuperation of the initially induced contractions. Prostaglandin F2alpha did not reverse the effect of diclofenac on agonist-induced contractions. Removal of diclofenac or NS-398 from the organ baths showed that the inhibition was totally reversible. Thus, the non-selective COX inhibitor diclofenac and the selective COX-2 inhibitor NS-398 are almost equipotent in reducing agonist-induced contractions in the isolated porcine ureter. Although the clinical relevance of this spasmolytic effect remains to be demonstrated, the data suggest that patients suffering from renal colic may benefit not only from the anti-diuretic and analgesic effects of diclofenac, but also from its potential spasmolytic properties. Moreover, selective COX-2 inhibitors may have clinical potential, as they may cause fewer side effects.