Non-length-dependent small fibre neuropathy. Confocal microscopy study of the corneal innervation.

BACKGROUND: It has been recently observed that small fibre neuropathy (SFN) may present as distal symmetrical polyneuropathy and with atypical non-length-dependent pattern. OBJECTIVE: To describe a small series of patients with non-length-dependent SFN, investigating corneal innervation with corneal confocal microscopy (CCM). METHODS: Evaluation of the corneal nerve fibre density using CCM in six women with non-length-dependent SFN. The patients were characterised by sensory disturbance involving proximal regions of the limbs, face and trunks, and the diagnosis was confirmed by the findings of decreased intraepidermal nerve fibre density on skin biopsy. RESULTS: Six women, aged 35-64, had non-length-dependent SFN, related to Crohn disease, impaired glucose tolerance and Sjögren's syndrome, or idiopathic (three cases). In all patients, CCM demonstrated decreased corneal nerve fibre density (12.5-23.4/mm(2); normal, >30.6/mm(2)). CONCLUSION: Non-length-dependent SFN may represent an intriguing diagnostic problem because of its puzzling presentation and the need for special investigations for its confirmation. In this perspective, CCM may provide a useful, non-invasive tool to complement the diagnostic workup.

Porcine sclera as a model of human sclera for in vitro transport experiments: histology, SEM, and comparative permeability.

PURPOSE: To evaluate porcine sclera as a model of human sclera for in vitro studies of transscleral drug delivery of both low and high molecular weight compounds. METHODS: Human and porcine scleras were characterized for thickness and water content. The tissue surface was examined by scanning electron microscopy (SEM), and the histology was studied with hematoxylin-eosin staining. Comparative permeation experiments were performed using three model molecules, acetaminophen as the model compound for small molecules; a linear dextran with a molecular weight of 120 kDa as the model compound for high molecular weight drugs; and insulin, which was chosen as the model protein. Permeation parameters such as flux, lag time, and permeability coefficient were determined and compared. RESULTS: Human and porcine scleras have a similar histology and collagen bundle organization. The water content is approx 70% for both tissues while a statistically significant difference was found for the thickness, porcine sclera being approximately twofold thicker than human sclera. Differences in thickness produced differences in the permeability coefficient. In fact, human sclera was found to be two to threefold more permeable toward the three molecules studied than porcine sclera. CONCLUSIONS: The results obtained in the present paper prove that porcine sclera can be considered a good model for human sclera for in vitro permeation experiments of both low and high molecular weight compounds. In fact, if the different tissue thickness is taken into account, comparable permeability was demonstrated. This suggests a possible use of this model in the evaluation of the transscleral permeation of new biotech compounds, which currently represent the most innovative and efficient therapeutic options for the treatment of ocular diseases.

A cytotoxicity study of silicon oxycarbide nanowires as cell scaffold for biomedical applications.

GOAL: Nanowires are promising biomaterials in multiple clinical applications. The goal of this study was to investigate the cytotoxicity of carbon-doped silica nanowires (SiOxCy NWs) on a fibroblastic cell line in vitro. MATERIALS AND METHODS: SiOxCy NWs were grown on Si substrates by CVD process. Murine L929 fibroblasts were cultured in complete DMEM and indirect and direct cytotoxicity tests were performed in agreement with ISO 19003-5, by quantitating cell viability at MTT and chemiluminescent assay. Cell cultures were investigated at Scanning Electron Microscope (SEM) and immunocytochemistry to observe their morphology and investigate cell-NWs interactions. Furthermore, hemocompatibility with Platelet-rich Plasma was assayed at SEM and by ELISA assay. RESULTS: SiOxCy NWs proved biocompatible and did not impair cell proliferation at contact assays. L929 were able to attach on NWs and proliferate. Most interestingly, L929 reorganised the NW scaffold by displacing the nanostructure and creating tunnels within the NW network. NWs moreover did not impair platelet activation and behaved similarly to flat SiO2. CONCLUSIONS: Our data show that SiOxCy NWs did not release cytotoxic species and acted as a viable and adaptable scaffold for fibroblastic cells, thus representing a promising platform for implantable devices.

Molecular genetics of autosomal dominant retinitis pigmentosa (ADRP): a comprehensive study of 43 Italian families.

Retinitis pigmentosa is the most common form of retinal degeneration and is heterogeneous both clinically and genetically. The autosomal dominant forms (ADRP) can be caused by mutations in 12 different genes. This report describes the first simultaneous mutation analysis of all the known ADRP genes in the same population, represented by 43 Italian families. This analysis allowed the identification of causative mutations in 12 of the families (28% of the total). Seven different mutations were identified, two of which are novel (458delC and 6901C-->T (P2301S), in the CRX and PRPF8 genes, respectively). Several novel polymorphisms leading to amino acid changes in the FSCN2, NRL, IMPDH1, and RP1 genes were also identified. Analysis of gene prevalences indicates that the relative involvement of the RHO and the RDS genes in the pathogenesis of ADRP is less in Italy than in US and UK populations. As causative mutations were not found in over 70% of the families analysed, this study suggests the presence of further novel genes or sequence elements involved in the pathogenesis of ADRP.

A novel dye exclusion method for testing in vitro chemosensitivity of human tumors.

Dissociated cancer cells are exposed to antineoplastic drugs (5 X 10(4) viable cells/drug for 1 hr or continuously) and cultured for 4 to 6 days in liquid medium. Cells are then stained with Fast green dye, sedimented onto slides with a Cytospin centrifuge, and counterstained with a modified hematoxylin and eosin technique. Dead cells stain with Fast green, and living cells stain with hematoxylin and eosin. Cell kill is calculated as percentage of control based on the relative numbers of living tumor cells, living non-tumor cells, and dead cells. Drug sensitivity could be assayed in 125 of 162 specimens of human neoplasms obtained from malignant effusions (16 of 18), excisional biopsies (31 of 44), needle biopsies (34 of 47), endoscopic biopsies (18 of 23), peripheral blood samples (19 of 20), and bone marrow aspirates (five of seven). The assays were successful (median of ten drugs tested) in: 46 of 64 adenocarcinomas; four of 11 squamous cell carcinomas; five of seven lymphomas; six of seven melanomas; two of four sarcomas; 18 of 20 transitional-cell carcinomas; 14 of 15 small-cell carcinomas; seven of eight myelomas; 12 of 12 chronic lymphocytic leukemias; seven of nine acute leukemias, and four of five "undifferentiated" carcinomas. The assay results demonstrated a strong correlation between the in vitro chemosensitivity of different types of tumors and the known clinical response patterns of these tumors. This assay can be used to determine which specific cells are killed in a heterogeneous cell population. Further work is needed to determine if this assay may be useful in blind screening trials for antineoplastic agents or if it may be of clinical use in predicting response to agents which are not cycle specific.

High rate of methicillin resistance of Staphylococcus aureus isolated from hospitalized nursing home patients.

Three series of studies were conducted between 1984 and 1986 at two community hospitals in Chicago to determine the frequency of methicillin resistance of Staphylococcus aureus isolated from hospitalized nursing home patients. Overall, 76 (49%) of 155 patients with S aureus clinical isolates admitted from 25 nursing homes had methicillin-resistant S aureus. In 1986, 53.1% of patients with S aureus infection or colonization among nursing home patients had the resistant strain, in contrast with 13.2% among patients from the community. A high incidence of methicillin-resistant S aureus infection or colonization is a clinical feature of nursing home patients. This observation may lead to an improvement in the clinical management of the patients suspected of having S aureus infection and alterations in the current policy of nursing homes that bar admission of patients carrying the resistant bacteria.

The study of visual evoked potentials in patients with thyroid-associated ophthalmopathy identifies asymptomatic optic nerve involvement.

In the present study we have recorded visual evoked cortical potentials (VECP) in 88 patients affected by autoimmune thyroid disease and thyroid-associated ophthalmopathy (TAO) without clinical signs of optic neuropathy. At the time of ophthalmological examination, 37 of these patients were hyperthyroid, 41 were euthyroid, and 8 were hypothyroid; 2 were not assessed. Twenty-nine normal subjects served as controls. We performed pattern reversal visual stimulation and recorded the amplitude and latency of the cortical electric response at 100 ms (P100 wave). There were no differences in the mean P100 amplitude of TAO patients and normal subjects. The mean P100 latency in patients was 105.6 +/- 0.5 ms, significantly higher than that in normal subjects (102.0 +/- 0.5 ms; P < 0.00003). Latency in euthyroid patients did not differ from that in either hypo- or hyperthyroid patients. The VECP test was positive (latency, > or = 110.0 ms) in 21 (23.8%) TAO patients. In patients with proptosis greater than 21 mm, latency was 106.7 +/- 0.7 ms, significantly higher than that in patients with normal Hertel measurements (104.3 +/- 0.6 ms; P < 0.01). Latency was not increased in patients with acute inflammatory signs compared to those with inactive eye disease and in patients with altered extrinsic motility. In patients with an abnormal visual field study, the mean latency was 110.3 +/- 1.5 ms, significantly higher than that in patients with a normal visual field (104.7 +/- 0.4; by t test, P < 0.000003). In conclusion, we observed a prolongation of the latency of the evoked cortical response in patients with TAO without subjective visual complaints and without optic nerve compression. We believe that the study of VECP in TAO is complementary to the study of the visual field in identifying early optic nerve dysfunction in the absence of decreased visual acuity.

Structural organization of the human gamma-glutamyl hydrolase gene.

Gamma-glutamyl hydrolase (GH) plays an important role in the metabolism of folic acid and the pharmacology of antifolates such as methotrexate. We have previously cloned and characterized the human GH cDNA. In this report, the complete organization and structure of the human GH gene was determined. The human GH gene spans 24 kb in the human genome, with nine exons sized from 51 to 371 bp. All of exon-intron splice junctions follow the GT-AG rule. The sequence upstream of exon 1 consists of a promoter-like, GC-rich region and a number of putative cis active elements including Sp1, AP1, and MZF1 sites. A TATA sequence in the 5' region of human GH gene was not observed, similar to housekeeping genes known to be tissue-specific and differentially expressed. S1 nuclease protection analysis with human liver, prostate, brain, and mammary gland revealed a major transcription start point at nucleotide -125 relative to the ATG start codon and several minor transcription start points. Analysis of GH cDNA isolated from human liver indicated a nucleotide change, T-->C, in the leader sequence of GH, which suggested a polymorphism. Studies of cDNA from different human tissue sources provided evidence that there is a single spliced cDNA species in human.

Changes in glucose level affect rod function more than cone function in the isolated, perfused cat eye.

The glucose concentration (gl) in mammalian serum incorporates a normal range of variation of several millimoles. We studied the effects of such variations on light-evoked electrical signals in the in vitro arterially perfused cat eye, avoiding extraocular regulatory mechanisms that might confound data interpretation. Changes in gl from the nominal control value of 5 mmol/l were maintained for 5-40 min. Stimuli of near rod threshold intensity were presented in full dark adaptation, and stimuli of higher intensity were presented in the presence of a white background for cone responses. We recorded the dc-electroretinogram (ERG), the scotopic threshold response (STR), the optic nerve response (ONR), and the transretinal slow P-III and transepithelial retinal pigment epithelium c-wave from the subretinal space. The ocular standing potential changed by up to +/- 2 mV in parallel with an increase and decrease in gl, independent of the adaptation condition. Our results show that the rod-ERG, STR, and rod-driven optic nerve response (ONR) have a marked sensitivity to small changes in gl (+/- 1 to 3 mmol/l). The field potentials increased and decreased in parallel with changes in gl. The cone ERG and cone ONR, in contrast, failed to respond consistently to increases in gl and revealed decreases in amplitudes only with an extreme decrease in gl. Decrease in gl, down to 2 mmol/l and less, is known to induce drastic behavioral and electrophysiologic phenomena in the central nervous system. Our results imply that the "normal" glucose level, at least in the cat, could be marginal for rod-mediated retinal function. The results also suggest a marked difference in metabolic mechanisms for cone versus rod photoreceptors.

Retroillumination versus reflected-light images in the photographic assessment of posterior capsule opacification.

PURPOSE: To investigate the relative merit of retroillumination and of reflected light slit-lamp-derived photographs in the assessment of the opacification of the posterior lens capsule. METHODS: Retroillumination and slit-lamp-derived reflected-light photographs were taken on 23 consecutive eyes with posterior capsule opacification (PCO) in uncomplicated pseudophakia. Subjective grading was performed on both types of photographs to evaluate the extent and density of posterior capsular opacification. Best-corrected visual acuity (BCVA) before and after YAG laser capsulotomy was used to assess the impact of capsular opacification on visual function. RESULTS: After capsulotomy all patients attained a BCVA > or = 46 letters (> or =20/32) with a mean increase of 25 letters, indicating that PCO was the cause of visual impairment in these patients. The relative capacity of retroillumination and of reflected-light photographs to adequately capture the extent and the severity of posterior capsule opacification varied considerably. Reflected-light images, in addition to frequently producing higher severity scores for the opacity than retroillumination photographs, in 4 of 23 eyes (17.4%) proved to be the only technique able to document the presence of PCO. CONCLUSIONS: Our results indicate that, with respect to retroillumination images, reflected-light photography has an increased ability to adequately capture the presence and the severity of PCO and that the use of only retroillumination images may lead to its underestimation. This may be relevant to clinical studies aiming to evaluate incidence and progression of this condition.

ETDRS-fast: implementing psychophysical adaptive methods to standardized visual acuity measurement with ETDRS charts.

PURPOSE: To measure visual acuity (VA) on Early Treatment Diabetic Retinopathy Study (ETDRS) charts with a modified faster procedure (ETDRS-Fast), based on adaptive psychophysics methods and to assess the method's validity and reproducibility. METHODS: Whereas the standard method for measuring VA with the ETDRS charts requires that the subject read all the letters beginning with the top row, in the ETDRS-Fast procedure, the subject is asked to read only one letter per row until a mistake is made. Then, following simple rules, the examiner finds a row from which the subject can begin reading all the letters downward, thus making the method identical with the standard method near threshold. VA determination was performed twice with both methods in 57 subjects in two separate sessions to assess validity and reproducibility. RESULTS: In both sessions the correlation between the two procedures was high (intraclass correlation coefficient 0.95), confirming the validity of the ETDRS-Fast procedure. Reproducibility was good for both procedures, with intraclass correlation coefficients of 0.94 for the standard and 0.96 for the ETDRS-Fast method. The ETDRS-Fast procedure allowed a significantly shorter test duration (-30%; P < 0.0001). CONCLUSIONS: Adaptive procedures allow accurate and fast determination of psychophysical thresholds by reducing the number of stimulus presentations when the subject is far from threshold. In the ETDRS-Fast method a few simple rules applied to optotype chart reading allow adaptation to each patient's level of VA. The ETDRS-Fast procedure significantly reduces test time and still yields results that are as accurate as those obtained with the standard method.

Confusion points and constant-luminance planes for trichromats, protanopes and deuteranopes.

The confusion points of dichromats are derived from the constant-luminance planes of trichromats, protanopes and deuteranopes experimentally defined by heterochromatic-flicker photometry: (1) the zero-luminance planes of the observers considered in this experiment intersect almost exactly in a line that crosses the plane of the chromaticity diagram in the tritanopic-confusion point and confirm that the short-wavelength sensitive cones can be considered to have no contribution to luminance; (2) protanopic- and deuteranopic-confusion points are taken as being defined by the intersection of the tangent line to the long-wavelength region of the spectrum locus and the zero-luminance plane for protanopes and deuteranopes, respectively.

Endoprosthesis implantation in the treatment of recurrent urethral stricture: a multicenter study. Sicilian-Calabrian Urology Society.

PURPOSE: This study was conducted by nine urology departments in southern Italy to assess the efficacy of and tolerance to treatment of recurrent urethral stricture using a permanent prosthesis. PATIENTS AND METHODS: Since 1992, 99 prostheses have been implanted to treat inflammatory and iatrogenic (seven departments) or all types (two departments) of urethral strictures. The Urolume Wallstent was used in 94 cases. Three centers implanted more than one prosthesis when this was indicated. Local anesthesia was used by six centers, spinal anesthesia by two, and local or general by one. At three centers, urethrotomy was performed immediately prior to implantation; two centers used dilation to 30F, and two centers performed urethrotomy 24 or 36 hours before implantation. The median follow-up is 29.1 months (range 3-53 months). RESULTS: The results were good in 52%, fair in 34%, and poor in 14% of patients. The maximum flow rate increased >75% in 82% of patients. All departments reported complete reepithelialization of the urethra by 6 months. The short-term complications (7-28 days) were perineal discomfort (86%) and dribbling (14%). The long-term complications were painful erection (44%), mucous hyperplasia (44%), recurring stricture (29%), and incontinence (14%). All departments performed resection for hyperplasia in many cases. CONCLUSION: Permanent urethral endoprostheses can produce excellent results in patients with recurrent urethral strictures.

[Hepatic metastases of colorectal carcinoma. Case reports]. / Metastasi epatiche da carcinoma colo-rettale. Note di casistica.

Authors report their clinical experience concerning liver metastases from colorectal cancer. 21 (26%) out of 87 patients observed between 1977 and 1988, underwent hepatic resection. In this group the recorded operative mortality was 4.7% and the postoperative follow-up showed a survival rate of 27% five years after hepatic surgery. Hepatic resection should be considered as the most effective treatment for liver metastases from large bowel cancer. Unfortunately only few patients are suitable for surgery. Alternative treatments for unresectable hepatic recurrences of colorectal cancer still remain a most major problem.

[The prevention of postoperative deep venous thrombosis with heparan sulfate per os. A controlled clinical study vs. heparin calcium]. / Prevenzione della trombosi venosa profonda postoperatoria con eparansolfato per os. Studio clinico controllato vs eparina calcica.

The paper reports the results of a randomised study carried out on a pool of 100 patients undergoing surgery in order to assess the efficacy and tolerability of orally administered heparan sulfate versus heparin calcium in the prevention of postoperative deep-vein thrombosis. Despite the limits of oral administration, heparan sulfate is indicated in the pharmacological prevention of postoperative deep-vein thrombosis.

Emotional reactions to music by gender, age, and expertise.

Fragments of classical music were submitted to 80 subjects, 40 children 9 to 10 years old and 40 adults 19 to 29 years old who were divided into eight groups of ten, to induce feelings of happiness, sadness, anger, and fear. The task required linking each piece of music to one emotion, identifying at the same time the intensity of the emotional response on a scale of 1 to 3. The goal was to study how gender, age, and exposure or expertise related to emotional perceptions of music. Analysis showed (a) experts in music and nonexperts ascribed similar emotions to pieces of music, (b) there was no difference in emotional response to music by gender, although women linked to music stronger emotions of anger than girls, (c) children perceived greater feeling of happiness in music and less feeling of anger than adults, and (d) emotions of anger and fear in music were often confused with one another.

[The effect of glucose on rod function in the isolated, perfused cat eye]. / Glukose-Wirkung auf die Stäbchenfunktion im isolierten, perfundierten Katzenauge.

Effects of changing glucose concentration on light-evoked, rod-matched electrophysiological responses were studied in isolated, arterially perfused cat eyes. 1. changes in glucose concentration in the perfusate induced remarkable effects in the electrical responses of retinal pigment epithelium neural retina, and optic nerve. 2. The ERG b-wave and the optic nerve response (ONR) were enhanced when glucose concentrations were increased above the standard level (5.5 mM), and were less than normal after termination of the increase in glucose, prior to recovery. 3. Decreasing glucose from the standard led to attenuation of b-wave and ONR. Both responses recovered completely upon returning to 5.5 mM glucose 4. When the control perfusate contained higher glucose concentrations (8-10 mM), additional glucose failed to elicit significant effects on the b-wave and on the ONR. 5. Increasing glucose induced a small transient decrease in standing potential, followed by a marked and maintained increase. Decreasing glucose induced changes of similar magnitude but opposite polarity. 6. Supply of glucose to mammalian retina in vitro is crucial for optimal sensitivity as shown in rod-matched signals from the perfused cat retina.

Functional modifications of endocrine pancreas after internal biliary fistula (experimental research).

The function of the B-insular system has been explored in dogs total internal biliary fistula (cholecysto-jejunostomy after closure of the common bile duct), an operation which causes an increase in number and total volume of pancreatic islets. Two groups of 10 animals each were either submitted to operation or used as controls. The endocrine function of the pancreas was explored 45-60 cays after surgery by assessing basal blood glucose and serum insulin. IVGTT and i.v. tolbutamide test were performed subsequently. The results indicate that in operated animals the ability to release insulin after stimulation is increased and that the control of basal hormone secretion is fully preserved.