Mouse antisera specific for desmosomal adhesion molecules of suprabasal skin cells, meninges, and meningioma.

Mouse polyclonal antisera were raised to the Mr 130,000 and Mr 115,000 cell surface glycoproteins, desmocollins, of desmosomes from bovine nasal epithelium. Immunoblotting confirmed that the antisera were specific for the desmocollins. An immunofluorescence study showed that the antisera distinguished between the basal and suprabasal layers of bovine and human epidermis. The antibodies reacted with cultured keratinocytes only after calcium-induced stratification. In epidermis, therefore, there appears to be a difference between the desmocollins of basal and suprabasal cells that may be important in relation to epidermal differentiation. Previous work has shown that polyclonal antisera raised in other animals (guinea pigs and rabbits) against desmocollins, as well as against other desmosomal components, react with all desmosome-containing epithelia. In contrast, an immunofluorescence survey of bovine, rat, and human tissues showed that the present mouse antisera stained only suprabasal skin cells and the arachnoid layer of the meninges, demonstrating that these have common determinants that distinguished their desmocollins from those of all other tissues. The antibodies also stained 11 of 12 meningiomas and, therefore, may be useful as a marker not only for the diagnosis of these tumors but also for investigation of their histogenesis.     

Selective photothermolysis: contribution to the treatment of flat angiomas (port wine stains) by laser

Since 1962, lasers have been used in dermatology and have become the first choice in the treatment of superficial, vascular ectasia. Lasers are unique sources of light; they are coherent, monochromatic, collimated and intense. By careful selection of wavelength, pulse duration, and intensity, it is often possible to selectively confine a laser effect to a specific histologic structure in tissue, depending upon the tissue properties. The ideal treatment of Port Wine Stains (PWS) should irreversibly damage the ectatic vessels but minimize heating of the epidermis and superficial dermis. A theory, called selective photothermolysis, predicts the optimal combination of laser parameters of achieving this ideal treatment of PWS to be a wavelength of 577 nm, a pulse duration of 0.35-10 msec, and an energy per surface area of about 7-8 J/cm2. Laser wavelength: The wavelength of 577 nm is preferred because it: maximizes the selective absorption by hemoglobin, minimizes absorption by epidermal melanin, provides sufficient depth of penetration in the blood to coagulate 0.1 mm vessels allows penetration of light into dermis up to 1 mm. Laser pulse duration: A pulse-width in the range of 0.35-10 msec allows the temperature elevation to be uniform inside the vessel and to be confined to the vessel area. Shorter pulses superheat the red blood cells causing explosive boiling and hemorrhage. Longer pulses allow heat to diffuse away from vessels, requiring greater energies per pulse to achieve vessel damage. An increased energy per pulse increases the risk of excessive damage to surrounding tissue.(ABSTRACT TRUNCATED AT 250 WORDS)     

Different mechanisms of decreased drug accumulation in doxorubicin and mitoxantrone resistant variants of the MCF7 human breast cancer cell line.

We selected two drug resistant variants of the MCF7 human breast cancer cell line by chronic in vitro exposure to doxorubicin (MCF7/D40 cell line) and mitoxantrone (MCF7/Mitox cell line), respectively. The cell lines are similar in growth characteristics including doubling time, DNA synthetic phase and cell size. Resistance to mitoxantrone conferred only partial resistance to doxorubicin; whereas resistance selected for doxorubicin appeared to confer complete resistance to mitoxantrone. Both agents selected for cross resistance to the Vinca alkaloids. MCF7/D40 cells display a classic-multi-drug resistance phenotype with expression of P-glycoprotein, decreased drug accumulation relative to the parental line and reversal of drug accumulation and drug resistance by verapamil. MCF7/Mitox cells likewise display resistance to multiple drugs, but in contrast to MCF7/D40 cells do not express P-glycoprotein by immunoblot or RNA blot analysis. Net drug accumulation in MCF7/Mitox cells was decreased relative to the parental cells but there was no selective modulation of drug accumulation or in vitro drug resistance by the addition of verapamil. Efflux of mitoxantrone was enhanced in both the MCF7/D40 and MCF7/Mitox cell lines relative to the MCF7/S cell line. We conclude that the two drug resistant cell lines have different mechanisms of decreased drug accumulation.     

Treatment of vasospasm with a 480-nm pulsed-dye laser

Laser energy at a wavelength of 480 nm was applied in 1-microseconds pulses of 3 to 10 mJ to two models of vasospasm. Rabbit common carotid arteries (CCA's) were constricted chronically by the application of human blood within a silicone sheath. Peak vasospasm developed 24 to 48 hours later, and persisted for up to 6 days. Endovascular laser treatment was delivered to 40 CCA's via a 200-microns diameter silica quartz fiber introduced through the femoral artery. The CCA caliber increased from 60% of the pre-vasospasm control diameter to a minimum post-laser diameter of 83% of control. No instances of laser-induced perforation or of arterial thrombosis were observed for up to 60 days after treatment. Prophylactic laser application to nine normal vessels was able to attenuate the development of vasospasm if blood was applied immediately thereafter (88% vs. 59% of control diameter, p less than 0.02), but not if blood was applied 7 days later. Studies in 16 normal CCA's established that there was a considerable margin between the laser energy required to induce dilatation and that which caused perforation, providing that the fiber remained relatively central within the artery. Morphological examination demonstrated focal loss of endothelial cells immediately after laser application, followed approximately 7 days later by the development of areas of intimal hyperplasia. Only minimal changes were observed in the medial or adventitial layers. In a second study, the basilar artery of seven dogs was constricted chronically by two intracisternal injections of autologous blood 3 days apart. Five dogs received endovascular laser treatment 7 or 10 days after the first injection, when basilar artery diameter was reduced to a mean of 61% and 77% of control, respectively. Immediately following treatment, basilar artery diameter increased to 104% and 102% of resting diameter, respectively. Both untreated and laser-treated arteries were smaller than the control diameter at 30 days (80% and 82%, respectively), but in each group the vasodilatory response to hypercapnia was preserved. These findings indicate that 1-microsecond laser pulses are well tolerated by systemic and cerebral arteries in two different animal models, and suggest that the 480-nm pulsed-dye laser may have an application for the treatment or prophylaxis of cerebral vasospasm.     

A method for evaluating systems of epidemiological surveillance

Epidemiological surveillance is the systematic collection, analysis and dissemination of health data for the planning, implementation and evaluation of public health programmes. Established surveillance systems should be regularly reviewed on the basis of explicit criteria of usefulness, cost and quality; systems should be modified as a result of such review. Attributes of quality include: (i) sensitivity, (ii) specificity, (iii) representativeness, (iv) timeliness, (v) simplicity, (vi) flexibility and (vii) acceptability. To date, evaluation of surveillance systems has been limited in scope and content. The evaluation method proposed in this article offers an organized approach to the evaluation of epidemiological surveillance systems. The usefulness of a surveillance system is measured by whether it leads to prevention or control or a better understanding of adverse health events. The measure can be qualitative, in terms of the subjective views of those using the system, or quantitative in terms of the impact of surveillance data on policies, interventions or the occurrence of a health event. The cost of a system includes indirect as well as direct costs, and should be measured in relation to the benefits obtained, such as reduction of medical-care expenses and of time lost from work. All elements of the system should be included in the cost: data collection, analysis and dissemination. The sensitivity of a surveillance system is its ability to detect health events (completeness of reporting). Its specificity is inversely proportional to the number of false positives it reports. Reports of a disease that do not meet the case definition are false positives, and may result in resources being wasted in investigating them.(ABSTRACT TRUNCATED AT 250 WORDS)     

MR of the cerebral operculum: abnormal opercular formation in infants and children.

PURPOSETo evaluate abnormalities of the cerebral operculum in infants and children and to propose the embryogenic basis of abnormal opercular formation as determined from MR imaging findings.METHODSEighty-six infants and children who had abnormally wide interopercular distances and/or distorted opercular topography seen on MR images were studied retrospectively. Clinically, patients presented with tonal abnormalities, macrocephaly, microcephaly, seizures, developmental delay, cerebral palsy, or facial dysmorphism. The abnormal opercula were compared with developing opercula at different stages of gestation.RESULTSAmong the 86 infants and children, two categories of opercular abnormalities were identified: an underdeveloped operculum (n = 64) and a malformed operculum (n = 22). The malformed operculum was further classified into three subtypes: nonformation of the operculum with lissencephaly (n = 1, 1%), abnormal opercular formation with pachygyria (n = 11, 13%), and nonformation or abnormal formation of the operculum without pachygyria or lissencephaly (n = 10, 12%). Two subtypes of the underdeveloped operculum were identified: an open operculum without a normal insula (n = 6, 7%) and an open operculum with a normal insula (n = 58, 67%). The five subtypes of abnormal opercular configuration showed a range of maturity that was comparable to the developing operculum at different ages.CONCLUSIONOpercular anomalies appear to follow sequentially predetermined normal steps in development. Arrest in opercular development or malformation may occur after an initial insult. MR imaging is the method of choice by which to identify these abnormalities.     

Myocardial perfusion and function in dogs with moderate coronary stenosis

MRI studies of first-pass contrast enhancement with polylysine-Gd-DTPA and myocardial tagging using spatial modulation of magnetization (SPAMM) were performed to assess the feasibility of a combined regional myocardial blood flow and 2D deformation exam. Instrumented closed-chest dogs were imaged at a baseline control state (Cntl) followed by two interventions: moderate coronary stenosis (St) achieved by partial occlusion of the left anterior descending (LAD) and moderate coronary stenosis with dobutamine loading (StD). Hypoperfusion of the anterior region (ANT) of the myocardium (LAD distribution) relative to the posterior wall (POS) based on the upslope of the signal intensity time curve from the contrast-enhanced MR images was demonstrated only with dobutamine loading (ANT:POS Cntl=1.077 ± 0.15 versus ANT:POS StD=0.477 ± 0.11, P<0.03) and was confirmed with radio-labeled microspheres measurements (ANT:POS Cntl=1.18 ± 0.2 ml/min/g versus ANT:POS StD=0.44 ± 0.1 ml/min/g; P<0.002). Significant changes in regional myocardial shortening were only seen in the StD state (P<0.02); the anterior region showed impaired myocardial shortening with dobutamine loading (P=NS), whereas the nonaffected POS region showed a marked increase in shortening when compared with Cntl (Cntl=0.964 ± 0.02 versus StD=0.884 ± 0.03; P<0.001). These results demonstrate that an integrated quantitative assessment of regional myocardial function and semiquantitative assessment of myocardial blood flow can be performed noninvasively with ultrafast MRI.     

Canine parvovirus capsid assembly and differences in mammalian and insect cells

We examined the assembly processes of the capsid proteins of canine parvovirus (CPV) in mammalian and insect cells. In CPV-infected cells empty capsids assembled within 15 min, and then continued to form over the following 1 h, while full (DNA-containing) capsids were detected only after 60 min, and those accumulated slowly over several hours. In cells expressing VP1 and VP2 or only VP2, empty capsid formation was also efficient, but was slightly slower than that in infected cells. Small amounts of trimer forms of VP2 were detected in cells expressing wild type capsid proteins, but were not seen for mutants containing changes that prevented capsid assembly. CPV capsids accumulated in the cell nucleus, but mutant VP1 and VP2 proteins that did not assemble became distributed throughout the nucleus and the cytoplasm, irrespective of whether they were expressed as VP1 and VP2, or as VP2 only. Urea or pH treatment of empty capsids released dimer, trimer, or pentamer capsid protein combinations, while treatment of full capsids consistently released trimer and, in some cases, pentamer forms. When wild type or assembly-defective VP2 genes were expressed from recombinant baculoviruses in insect cells, most of the protein was recovered as noncapsid aggregates, and only a small proportion assembled into capsids. Both the assembled capsids and the noncapsid aggregates were seen primarily in the cytoplasm of the insect cells. The VP2 expressed in insect cells that was recovered in aggregates had an isoelectric point of about pH 6.3, while that recovered from assembled capsids had a pI of about 5.2, similar to that seen for the VP2 of capsids recovered from mammalian cells.     

Monoclonal antibody to desmosomal glycoprotein 1--a new epithelial marker for diagnostic pathology

Desmosomes are intercellular adhesive junctions that occur in almost all epithelia and should therefore be useful as epithelial markers in tumour diagnosis. Here, we describe a monoclonal antibody, 32-2B, to a major desmosomal glycoprotein (dgl) which reacts with human tissues in paraffin sections. This antibody was tested for its ability to stain epithelia and tumours. It reacted with all epithelia tested and with every specimen of a wide range of carcinomas. It also stained meningiomas, another desmosome-containing tumour. It did not stain other types of tumours including lymphomas, melanomas, and various sarcomas, or normal tissues which lack desmosomes. These characteristics demonstrate that 32-2B is a reliable epithelial marker that may have a useful role in diagnostic histopathology.