The presence of occipital hair in the pilonidal sinus cavity-a triple approach to proof.

PURPOSE: Hair in the pilonidal sinus is not growing within the sinus cavity, as hair follicles are not present there. Not few pilonidal patients do not have intergluteal hair, which is said to be the causative agent of folliculitis and pilonidal genesis. So, what is the real source of the hair forming the typical pilonidal hair nest? METHODS: A trifold approach was used: First, axial hair strength testing of pilonidal hair and body hair harvested from head, lower back (glabella sacralis), and cranial third of intergluteal fold. Hair strength match was compared clinically. Second, comparative morphological examination by expert forensic biologist of hair from sinus and dorsal body hair. Third, statistical Bayesian classification of every single sinus hair based on its strength was done to determine the most probable region of origin. RESULTS: Using clinical hair strength comparison, in 13/20 patients, head hair is the stiffest hair, followed by intergluteal hair. Only in 6/20 patients, this is the case with hair from the glabella sacralis. According to comparative morphological comparison, a minimum of 5 of 13 hair nests with possible hair allocation examined contain hair from the occiput. In 5/18 nests, hair could not be determined to a specific location though. Statistical classification with correction for multiple testing shows that 2 nests have hair samples that are at least 100 times more probable to originate from head or lower back than from intergluteal fold. CONCLUSION: We saw our null hypothesis that "hair in the sinus cavity is from the intergluteal region" rejected by each of three different approaches. There is strong evidence that occipital hair is present regularly in pilonidal sinus nests. We should start thinking of occipital hair as an important hair source for the development of the pilonidal hair nest.

Expression of extra domain A fibronectin sequence in vascular smooth muscle cells is phenotype dependent.

Different fibronectin (FN) variants arise from the single gene transcript alternatively spliced in a tissue-specific manner (Hynes, R. O. 1985. Annu. Rev. Cell Biol. 1:67-90; Owens, R. J., A. R. Kornblihtt, and F. E. Baralle. 1986. Oxf. Surv. Eurcaryotic Genes. 3:141-160). We used mAb IST-9, specific for extra domain A (ED-A) FN sequence, and cDNA probe to ED-A exon to determine whether ED-A is present in FN synthesized by vascular smooth muscle cells (SMCs) and, if so, whether expression of ED-A is SMC phenotype dependent. ED-A-containing FN (A-FN) was not revealed in tunica media of human arteries and normal rat aorta by immunofluorescence and immunoblotting techniques. A cDNA probe to ED-A exon did not hybridize with RNA isolated from human aortic media. A positive reaction with IST-9 was observed in (a) diffuse intimal thickening and atherosclerotic plaque from human arteries; (b) experimentally induced intimal thickening in rat aorta; and (c) cultured vascular SMCs. A-FN mRNA was present in the RNA preparation from human aortic intima as judged by hybridization with cDNA probe to ED-A. On the other hand, an mAb interacting with an epitope common for all FN variants revealed FN in both intima and media of human arteries and in the normal rat aorta. A cDNA probe to a sequence shared by all FN variants hybridized with RNA from both intima and media of human aorta, though the level of expression was higher in intima. The data suggest that ED-A exon is omitted during splicing of the FN mRNA precursor in medial SMCs while the expression of A-FN is characteristic of "modulated" SMCs--those of intimal thickenings, of atherosclerotic lesions, and growing in culture.

Inhibition of the action of nonsuppressible insulin-like activity on isolated rat fat cells by binding to its carrier protein.

Nonsuppressible insulin-like activity extracted and purified from human serum (NSILA-S) mimics all insulin-like effects in vitro and, after injection, in vivo in the presence of excess insulin antibodies. However, there is no evidence that it exerts acute insulin-like effects in its native form in the circulation, where it is almost completely bound to a specific large molecular weight carrier protein. In this paper we show that partially purified NSILA-S-carrier protein, devoid of endogenous insulin-like activity, inhibits the stimulatory effect of NSILA-S, but not of insulin, on 3-0-methylglucose transport and on lipogenesis from [U-(14)C]glucose in isolated rat fat cells. Concomitantly, it prevents binding of (125)I-labeled NSILA-S to the insulin receptor and to the NSILA-S-binding site. The following explanation is, therefore, offered for the absence of acute insulin-like effects of native NSILA-S in vivo: In native serum NSILA-S occurs almost exclusively as NSILA-S-carrier complex. According to recent findings the passage of this complex through blood capillaries is restricted. The present results indicate that, in addition, it is metabolically inactive, or, at least, possesses reduced metabolic activity. The well-known phenomenon that whole serum, nevertheless, exerts pronounced nonsuppressible insulin-like effects on adipose tissue in vitro seems, therefore, to be mainly caused by the presence of a large molecular weight insulin-like protein not identical to the NSILA-S-carrier complex.

Effect of systemic hypertension on foveolar choroidal blood flow in age related macular degeneration.

AIM: To investigate the effect of systemic hypertension (SH) on the foveolar choroidal circulation in patients with age related macular degeneration (AMD). METHODS: This study included 163 study eyes with early AMD characteristics of 124 AMD patients. Study eyes had visual acuity of 20/40 or better, drusen > or =63 microm, and/or RPE hypertrophy. 56 of the AMD patients had a history of SH and 47 of these patients were receiving antihypertensive medications. Laser Doppler flowmetry (Oculix) was used to assess relative choroidal blood velocity (ChBVel), volume (ChBVol), and flow (ChBFlow) in the centre of the fovea of the study eyes. Differences in the mean haemodynamic parameters between groups of eyes were assessed using analysis of variance (ANOVA) and a test of linear trend, with adjustment for the correlation between eyes of the same patient. RESULTS: AMD patients with SH showed decreased ChBFlow in comparison with those without SH (ANOVA, p = 0.02). This association was maintained after adjustments for multiple factors (p = 0.04). CONCLUSIONS: AMD patients with SH have lower ChBFlow than those without SH. This decrease in choroidal blood circulation may help explain the mechanism by which systemic hypertension may contribute to the progression of AMD and the development of choroidal neovascularisation.

Effect of niacin on retinal vascular diameter in patients with age-related macular degeneration.

PURPOSE: Niacin is a B vitamin well-known for causing vasodilation and flushing. The purpose of this study was to investigate its effect on the retinal vasculature of patients with age-related macular degeneration (AMD). METHODS: Twelve patients with AMD were enrolled in a double-blind, randomized, placebo-controlled, crossover trial. Fundus photographs of the posterior pole were taken at baseline, 30 min, and 90 min after a single dose of niacin or placebo. The protocol was repeated after a washout period using the alternate study drug. The diameters of two veins and one artery on each image were measured. RESULTS: An analysis of variance for repeated measures comparing the effects of niacin with those of placebo demonstrated a significant increase in the inferior temporal retinal artery diameter (p = 0.01), with a 5.3 +/- 7.7% increase at 30 min (p = 0.05) and 5.8 +/- 5.0% increase at 90 min (p = 0.003). No significant changes were observed in the temporal retinal veins. CONCLUSIONS: Our results suggest that niacin produces vasodilatation of retinal arterioles. Further studies are needed to ascertain whether niacin treatment may be beneficial in retinal ischemic diseases.

Immobilized bovine lactose synthase. A method of topographical analysis of the active site.

Bovine galactosyltransferase (UDPgalactose: D-glucose 4beta-galactosyltransferase, EC 2.4.1.22) was covalently coupled to Sepharose 4B by reaction at pH 5.0 with the activated mixed disulfide Sepharose-glutathione-2(5-nitropyridyl)-disulfide. The Sepharose-protein conjugate was presumably coupled via the unique highly reactive cysteine of those thiols on the bovine enzyme. The gel-bound N-acetyllactosamine and lactose synthase activity of about 0.4% was consistent with the affects of diffusion and the 90% activity reduction noted upon thiol modification of the dissolved enzyme. The residual lactose biosynthetic activity of the bound enzyme appeared possible only if the reactive thiol were physically distinct from the active site since the bulky Sepharose-glutathione group must not obscure the alpha-lactalbumin binding region.

Diabetic glycemic control and retinal blood flow.

The effect of strict glycemic control on retinal volumetric blood flow rate (Q) was investigated in 13 insulin-dependent diabetic patients with laser Doppler velocimetry and monochromatic fundus photography. Strict glycemic control was achieved by glucose monitoring and four daily insulin injections. Q was determined in a major retinal vein at baseline and then 5 days, 2 mo, and 6 mo after the institution of strict control. Level of retinopathy was assessed from stereocolor fundus photographs taken at baseline and 6 mo. After 6 mo of strict diabetic control, five eyes demonstrated progression (P) by one or more retinopathy levels, and eight eyes showed no progression (NP). At 5 days, there was a significant decrease in Q of 1.4 +/- 0.9 microliters/min (P less than 0.005) in NP eyes and a nonsignificant increase in Q of 1.2 +/- 1.7 microliters/min in P eyes. Changes in Q from baseline observed at 5 days were strongly correlated with changes in retinopathy level at 6 mo (r = 0.79, P less than 0.005). No significant changes in Q from baseline were observed at 2 and 6 mo. A lack of decrease in Q at 5 days was associated with the progression of retinopathy that occurs in some patients after the institution of strict glycemic control and may serve as a predictor for progression of retinopathy.

A correlative study on the topographical distribution of the receptors for low density lipoprotein (LDL) conjugated to colloid gold in cultured human skin fibroblasts employing thin section, freeze-fracture, deep-etching, and surface replication techniques.

The topographical distribution of the receptors for low-density lipoprotein (LDL) conjugated to colloidal gold on cultured human skin fibroblasts was studied using a surface replication technique following critical point drying. In this correlative study, also employing thin sectioning, freeze-fracture and deep-etching techniques in conjunction with the polyene antibiotic filipin, it could be shown that LDL preferentially binds to certain microdomains of the plasma membrane, the so-called coated pits. The coated pits can be recognized after filipin treatment both on the P face and E face of the membrane, due to the fact that they are free of filipin-sterol complexes. Filipin was also of value in clearly delimiting the structure of nascent coated pits from their surroundings in surface replicas. Using filipin has the effect of increasing the contrast between the smooth surface of a coated pit and the rough surface of the remaining membrane caused by formation of filipin-sterol complexes. This study has shown that there are differences in the topographical distribution and number of gold particles from fibroblast to fibroblast and even between different areas of the same cell. The surface replication technique using goldlabelled LDL provides s suitable method for improving the interpretation of the spatial arrangement of the LDL receptors. The advantages of this technique are discussed in comparison with the methods previously employed to visualize LDL receptors.

Effects of hypertension on migration and proliferation of smooth muscle in culture.

Migratory and proliferative characteristics of explanted rat aortic smooth muscle cells were studied in response to hypertension induced by 4 weeks of deoxycorticosterone-salt administration. Under low serum conditions (0.1% fetal bovine serum), over 80% of aortic medial explants from hypertensive rats yielded smooth muscle cell colonies after 8 days of culture while fewer than 10% of the control explants were positive. Time lapse video analysis of subsequent growth in the presence of 10% serum revealed that interdivision times of smooth muscle cells from hypertensive animals were significantly shorter than those in controls (p less than 0.01). Significant differences in proliferative capacity of smooth muscle cells were evident, even after one subculture (p less than 0.01). Comparison of these results with data from mechanical injury suggests that 4 weeks of deoxycorticosterone-salt hypertension can potentiate subsequent smooth muscle cell migration and growth in vitro to an extent similar to that observed with the combined effects of total endothelial denudation and wall distention by a balloon catheter.

[35S]proteoglycan metabolism of arterial smooth muscle cells cultured from normotensive and hypertensive rats.

Arterial smooth muscle cells cultured from normotensive and hypertensive rats incorporated [35S]sulfate into the extracellular and pericellular sulfated proteoglycans and endocytose extracellular [35S]proteoglycans at a significantly higher rate in the phase of logarithmic growth than did nondividing cells. 35S incorporation into proteoglycans was positively correlated with [3H]thymidine incorporation into the cellular TCA-precipitable material. The rates of [35S]proteoglycan synthesis and endocytosis per cell pr day were higher in smooth muscle cells from hypertensive than from normotensive animals, the observed differences being related to a higher average protein content of smooth muscle cells cultured from hypertensive rats as compared with cells of normotensive animals. Gel filtration under dissociative conditions separated the [35S]proteoglycans into high and low molecular weight fractions (A, B) differing in glycosaminoglycan composition and their ability to be endocytosed by smooth muscle cells. The relative proportion of the high molecular weight proteoglycan fraction A decreased continuously from sparse to confluent cell cultures.

Smooth muscle cell migration and proliferation: atherogenic mechanisms in hypertension.

The proliferative and migratory behavior of explanted rat aortic smooth muscle cells (SMC) was investigated in cells obtained from either 24-week-old normotensive Wistar-Kyoto (WKY) or age-matched spontaneously hypertensive (SHR) rats. Time lapse video analysis of primary SMC growth in the presence of 10% serum revealed that interdivision times of cells from SHR were significantly shorter than those from WKY. Differences in the proliferative capacity of these cells were still present after two subcultivations, as analyzed by conventional growth curves. In contrast to the proliferative behavior, no differences in the migratory characteristics of SMC could be detected in a migration assay analyzing the SMC outgrowth of standardized aortic explants under low serum conditions (0.1% fetal bovine serum). It has been shown that another model of hypertension, the 4 week DOC/salt hypertensive rat results in a different reaction of SMC. Therefore, it can be considered that the extent of the potentially atherogenic alterations of SMC function in hypertension is dependent on the type, duration and the rate of increase of hypertension.

Effect of timolol maleate on the retinal circulation of human eyes with ocular hypertension.

We studied the effect of topical timolol maleate 0.5% on the retinal circulation of eyes with ocular hypertension using laser Doppler velocimetry and monochromatic fundus photography. Patients with ocular hypertension had normal eye examinations and had documented elevated intraocular pressures of 23 mmHg or higher on two or more separate occasions. In a double-masked randomized design, one eye of each subject received timolol maleate 0.5% and the fellow eye received placebo. Vessel diameter, maximum velocity of red blood cells, and volumetric blood flow rate were determined in a major retinal vein of each eye just prior to the instillation of drops, and then 2 hr later. In comparison to the baseline value, there was an increase of 12.0% in average red blood cell velocity (P less than 0.005, statistically significant) and of 8.4% in volumetric blood flow rate in the timolol-treated eyes (P less than 0.05, statistically significant). No significant changes in these quantities were observed in the placebo-treated eyes. Also, no significant change in venous diameter was detected in the placebo- and the timolol-treated eyes. In comparison to the baseline, a significantly larger increase in red blood cell velocity was observed in the timolol-treated eyes than in the placebo-treated eyes (P less than 0.05). The difference between the increase in blood flow observed in the timolol-treated eyes and the placebo-treated eyes achieved a probability value of P = 0.058. The increase in blood flow observed in the timolol-treated eyes may be related to the increase in perfusion pressure produced by this drug.

Effect of topical timolol on the human retinal circulation.

The effect of topical timolol maleate 0.5% on the retinal circulation was investigated in 14 normal subjects using laser Doppler velocimetry and monochromatic fundus photography. In a double masked, randomized design, one eye received two drops of timolol maleate 0.5% and the fellow eye received two drops of placebo. Vessel diameter, maximum velocity of red blood cells, and volumetric blood flow rate were determined in a major temporal vein of each eye just prior to the instillation of drops, and then 90 min later. In comparison to the baseline value, there was a significantly larger average percentage increase in maximum velocity of red blood cells and volumetric blood flow rate in the timolol-treated eyes (11.0% and 13.2%, respectively) than in the control eyes (2.2% and 1.5%, respectively, paired t-test, P less than 0.05). No significant change in venous diameter was detected. No significant linear correlations were found between the percentage change in maximum velocity of red blood cells and mean brachial artery blood pressure, and between the percentage change in volumetric blood flow rate and mean brachial artery pressure in the placebo-treated eyes, whereas significant correlations were present between these quantities in the timolol-treated eyes (P less than 0.01 and P less than 0.05, respectively). The lack of such correlations in the control eyes is probably due to autoregulation of the retinal circulation. The presence of correlations in the timolol-treated eyes suggests that the drug may affect the capacity of the retina to autoregulate.

Effect of two weeks of timolol maleate treatment on the normal retinal circulation.

The effect of 2 weeks of topical treatment with timolol maleate 0.5% ophthalmic solution on the retinal circulation was investigated using bidirectional laser Doppler velocimetry and monochromatic fundus photography. Fifteen normal healthy volunteers were included in this study. In a double-masked, randomized design one eye of each received one drop of timolol maleate 0.5% twice daily for 14 days, and the fellow eye received placebo. Vessel diameter (D), maximum erythrocyte velocity (Vmax), and volumetric blood flow rate (Q) were determined in one major retinal vein of each eye before treatment, and then, 2 hr after the instillation of drops on the morning of the 15th day of treatment. After treatment, the average change from baseline in D (+0.7%), Vmax (-8.2%) and Q (-7.7%) were not statistically significant in the placebo-treated eyes. In the timolol-treated eyes, the average increase from baseline in D (+0.1%) and Q (+10%) were not statistically significant. Average Vmax, on the other hand, increased significantly from baseline (P less than 0.05) by 9.6%. In comparison to the placebo-treated eyes, Vmax and Q were significantly increased in the timolol-treated eyes (P less than 0.0005 and P less than 0.01, respectively). These results are similar to those reported previously in a study of the effect of a one-time instillation of timolol and, therefore, suggest that the effect of timolol on the retinal circulation is maintained over a 2-week period.

Effects of oxygen and carbon dioxide on human retinal circulation.

PURPOSE: Carbogen, a gas mixture of 95% O2 and 5% CO2, is given to patients with retinal artery obstruction in an attempt to improve retinal oxygenation. The purpose of this study was to compare the effects of carbogen and 100% O2 breathing on retinal blood flow. METHODS: On two separate occasions, 12 normal, healthy volunteers breathed air and then either 100% O2 or carbogen while laser Doppler velocimetry measurements and monochromatic fundus photographs were taken. Retinal vessel diameter, maximum velocity of red blood cells, and volumetric blood flow rate were determined in a main temporal vein. RESULTS: Both 100% O2 and carbogen caused significant average reductions in vessel diameter (14.1% and 10.6%, respectively), maximum red blood cell velocity (42.1% and 27.3%, respectively), and blood flow (56.4% and 42.2%, respectively). The average vasoconstriction of the large retinal veins caused by carbogen was not significantly smaller than that caused by 100% O2. The average reductions in maximum red blood cell velocity and blood flow caused by carbogen were significantly smaller than those caused by 100% O2 (P < .001 and P < .01, respectively). CONCLUSIONS: In normal subjects, inhalation of carbogen leads to less reduction in blood flow than inhalation of 100% O2, presumably by reducing the vasoconstriction of small arterioles induced by elevated oxygen levels.

The acute effect of oral acetazolamide on macular blood flow.

Acetazolamide has been shown to be beneficial in the treatment of macular edema. To investigate whether this effect is associated with changes in the retinal circulation, the acute effect of oral acetazolamide on macular blood flow was studied in 20 healthy volunteers. The blue-field simulation technique, a noninvasive method enabling the quantitation of the number (N) and mean velocity (Vm) of leukocytes flowing in the subject's own macular capillaries was used in this study. On two different occasions, separated by 3 or more days, 20 subjects adjusted Vm and N of computer-simulated leukocytes moving on a video screen to match those of their own entoptically perceived leukocytes before and 3 hr after a double-blind, randomized administration of 500 mg acetazolamide or placebo capsules. Ten trials were done, and the velocities were averaged. After acetazolamide ingestion, there was a nonsignificant average change from baseline in Vm (2.5 +/- 23% [+/- one standard deviation]; P greater than 0.1, by paired student t-test) and N (6.9 +/- 25%, P greater than 0.1). After placebo ingestion, the average changes from baseline in Vm and N also were not statistically significant (-1 +/- 18% and 14.9 +/- 30.3%, respectively). Furthermore, when compared with the changes measured after placebo intake, acetazolamide ingestion was associated with a nonsignificant 4.3 +/- 28.7% change in Vm (P greater than 0.1) and a -8 +/- 30.9% change in N (P greater than 0.1). With 20 subjects tested, the calculated average minimum change in leukocyte velocity that could have been detected with this technique (P less than 0.05, by paired student t-test) is about 9%.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of topical carteolol on the normal human retinal circulation.

The effect of topical carteolol 1%, a beta-adrenergic blocker with intrinsic sympathomimetic activity, on the retinal circulation was investigated in 15 normal subjects using laser Doppler velocimetry and monochromatic fundus photography. In a double-masked randomized design, one eye received one drop of carteolol 1% and the fellow eye one drop of placebo. Vessel diameter, maximum erythrocyte velocity, and volumetric blood flow rate were determined in a major temporal vein of each eye just before instillation of the drops and then 120 min later. No significant changes in heart rate or mean brachial artery blood pressure were detected after treatment. Intraocular pressure decreased by 28% in the carteolol-treated eye (P less than 0.0001) and by 15% in the placebo-treated eye (P less than 0.001). No significant changes in vessel diameter, maximum erythrocyte velocity, and volumetric blood flow rate were observed in the carteolol-treated eyes (0.3%, 4.3%, and 3.6%, respectively) or the placebo-treated eyes (0.5%, 5.8%, and 6.7%, respectively).

Effect of topical timolol maleate on the ophthalmic artery blood pressure.

The effects of topical timolol maleate 0.5% on the ophthalmic artery diastolic (OABPd), systolic (OABPs) and mean blood pressure (OABPm) were investigated in 19 healthy subjects using compression ophthalmodynamometry. In a randomized, double-blind study, one eye of each subject received one drop of timolol maleate 0.5% and the fellow eye received a drop of placebo. Measurements of OABPd and OABPs were performed just prior to instillation of the drops, and then 2 hr later. OABPm was calculated as OABPm = OABPd + 1/3 (OABPs - OABPd). No significant changes in OABPd, OABPs or OABPm were observed following the drops in the timolol-treated eyes or in the placebo-treated eyes. A significant difference (P = 0.0198) was found, however, when the changes in OABPm occurring in the timolol-treated eyes were compared with the changes occurring in the placebo-treated eyes, suggesting that the drug may have an effect upon ophthalmic artery pressure and retinal artery pressure that is expressed differently in each eye.

The effect of caffeine on the human macular circulation.

The acute effect of caffeine on the retinal circulation was studied in 14 healthy volunteers using the blue field simulation technique, which provides measurements of the velocity of leukocytes flowing within the macular capillaries. Subjects adjusted the mean velocity (Vm) of computer-simulated leukocytes moving on a cathode ray tube screen to match that of their own entoptically perceived leukocytes before and 1 hr after a double-masked, randomized administration of 200 mg caffeine or placebo. Caffeine produced an average 13% +/- 5% (SEM) decrease in Vm (P less than 0.05) and a 9% +/- 3% increase in diastolic blood pressure (P less than 0.05). The decrease in Vm and, presumably, blood flow occurring despite the increased diastolic blood pressure probably is attributable to retinal vasoconstriction. This effect may result from caffeine's known inhibitory effect on adenosine, a potent vasodilator of the retinal vasculature.

Autoregulation of human retinal blood flow. An investigation with laser Doppler velocimetry.

The effect of acute changes in mean retinal perfusion pressure, P (2/3 of mean brachial artery blood pressure minus IOP), on retinal volumetric blood flow rate, Q, was investigated in normal volunteers. Changes in Q were determined from Q = k X Vmax X D2, where Vmax is the center line red blood cell velocity measured from temporal veins by laser Doppler velocimetry, D is the vessel diameter obtained by monochromatic fundus photography, and k is a constant of proportionality. A suction cup was used to induce step changes in IOP and, consequently, in P. The magnitude of the steps ranged from 10-32 mmHg. During the first 30 sec after a step decrease in P, Vmax and Q were significantly smaller than at rest by an amount proportional to the decrease in P. Thereafter, Vmax and Q increased markedly towards their values at rest, although P changed comparatively little during this period of time. Time constant of the corresponding decrease in vascular resistance, R(t) = P(t)/Q(t), was approximately 45 sec. There was no significant change in D during elevated IOP. Removal of the cup induced an immediate step increase in P, Vmax, D, Q, and R. Thereafter, Vmax, D, Q, and R returned to their values at rest (time constant of the change in R was about 30 sec), while P remained nearly constant. The rapid change in vascular resistance following a step decrease and increase in P can be attributed to an active process that attempts to maintain blood flow close to normal, in spite of changes in perfusion pressure (autoregulation).(ABSTRACT TRUNCATED AT 250 WORDS)