Measurement of the motility of endothelial cells in confluent monolayers.

OBJECTIVE: The reported experiments were designed to develop and test a method to measure the motility of endothelial cells in confluent cultures. METHODS: Endothelial cell cultures were recorded by time-lapse video. Frames from the recording were digitized, and cell locations were identified at 30-minute intervals using either the centroids or nuclei. The speed of the cells was determined for distances between their successive positions. Cells were stained immunocytochemically to determine the status of their adherens junctions. RESULTS: Prior to reaching confluence cells exhibited motility unhampered by collisions. After reaching confluence, cells continued to exhibit motility that decreased with time until the cells ceased effective movement. The motility in confluent monolayers persisted in the presence of complete adherens junctions. Motility could be restored in quiescent confluent cultures by the addition of fibroblast growth factor 2 (FGF-2). Generally, cells responding to FGF-2 moved without obvious loss of adherens junctions. CONCLUSION: These experiments provide evidence that (1) endothelial cells in culture are motile, (2) they do not form circumferential adherens junctions until the culture reaches confluency, (3) the establishment of adherens junctions itself does not interdict cell motion, and (4) the immotile state is reversible by FGF-2 in the presence of adherens junctions.

Cerebral mucormycosis: proton MR spectroscopy and MR imaging.

Proton magnetic resonance spectroscopy (MRS) was integrated with magnetic resonance imaging (MRI) in the evaluation of a case of cerebral mucormycosis. MRS showed markedly elevated lactate, depleted N-acetyl aspartate and metabolite resonances attributable to succinate and acetate. The spectroscopy profile is essentially similar to that of bacterial abscess but without the commonly seen resonances of the amino acids valine, leucine and isoleucine. Our extensive literature review did not yield any reports of MRS findings on cerebral mucormycosis. MRS prospectively limited the differential diagnoses given the otherwise nonspecific and complex MR imaging findings in our immunosuppressed patient.

Angiotensin II induces vascular smooth muscle cell replication independent of blood pressure.

The purpose of this investigation was to evaluate the role of blood pressure in the proliferative response of vascular smooth muscle cells to systemic infusion of angiotensin II (Ang II). Our laboratory has previously shown that infusion of Ang II induces smooth muscle cell proliferation in rat mesenteric vessels and carotid arteries. Ang II, a strong vasopressor, raised systolic blood pressure in rats from 120 to 200 mm Hg at a dose of 435 ng x kg(-1) x min(-1) after 1 week of treatment. The question arises as to whether this development of hypertension is a primary contributor to the replicative activities observed in the arterial wall of the mesenteric arteries or the carotid arteries or whether Ang II alone, without an increase in blood pressure, is sufficient to stimulate proliferation in these vessels. In the previous studies, we found that Ang II stimulated smooth muscle cell replication in the carotid artery and in type III and type I mesenteric microvessels. This study demonstrates that although administration of hydralazine normalizes the animals' blood pressures, it does not suppress the mitogenic effect of Ang II. Thus, it appears that Ang II has a direct effect on cell proliferation.

Activation of visuomotor systems during visually guided movements: a functional MRI study.

The dorsal stream is a dominant visuomotor pathway that connects the striate and extrastriate cortices to posterior parietal areas. In turn, the posterior parietal areas send projections to the frontal primary motor and premotor areas. This cortical pathway is hypothesized to be involved in the transformation of a visual input into the appropriate motor output. In this study we used functional magnetic resonance imaging (fMRI) of the entire brain to determine the patterns of activation that occurred while subjects performed a visually guided motor task. In nine human subjects, fMRI data were acquired on a 4-T whole-body MR system equipped with a head gradient coil and a birdcage RF coil using a T2*-weighted EPI sequence. Functional activation was determined for three different tasks: (1) a visuomotor task consisting of moving a cursor on a screen with a joystick in relation to various targets, (2) a hand movement task consisting of moving the joystick without visual input, and (3) a eye movement task consisting of moving the eyes alone without visual input. Blood oxygenation level-dependent (BOLD) contrast-based activation maps of each subject were generated using period cross-correlation statistics. Subsequently, each subject's brain was normalized to Talairach coordinates, and the individual maps were compared on a pixel by pixel basis. Significantly activated pixels common to at least four out of six subjects were retained to construct the final functional image. The pattern of activation during visually guided movements was consistent with the flow of information from striate and extrastriate visual areas, to the posterior parietal complex, and then to frontal motor areas. The extensive activation of this network and the reproducibility among subjects is consistent with a role for the dorsal stream in transforming visual information into motor behavior. Also extensively activated were the medial and lateral cerebellar structures, implicating the cortico-pontocerebellar pathway in visually guided movements. Thalamic activation, particularly of the pulvinar, suggests that this nucleus is an important subcortical target of the dorsal stream.

Histologic differences in benign prostate hyperplasia between Chinese and American men.

BACKGROUND: This investigation sought to determine morphologic differences between Chinese and Americans with symptomatic benign prostatic hyperplasia (BPH), in an admixture of stroma, epithelium, and luminal spaces. METHODS: Adjacent sections of simple prostatectomy specimens from China and the U.S. were stained to highlight the stroma and epithelium. Image analysis was performed on random fields. RESULTS: Chinese tissue had higher glandular densities (mean = 12.5 acini/mm2 vs. 6.2 acini/mm2; P < 0.0001), while American samples had higher percent stroma (mean = 66% vs. 51%; P = 0.0003). Mean luminal cell heights in the Chinese and American prostate acini were 11.7 and 19.0 microns, respectively (P < 0.0001). CONCLUSIONS: These data indicate significant histologic variation in BPH between Chinese and American men undergoing simple prostatectomy. These differences may provide a focus for investigating the epidemiological variation in clinical BPH and carcinoma between our two countries, and could have applications in clinical management of symptomatic BPH.

Serum basic fibroblast growth factor in men with and without prostate carcinoma.

BACKGROUND: Angiogenesis is essential for the growth of neoplasms. Increased vascularity has been associated with human prostatic carcinoma stage and has been shown to offer prognostic information. Basic fibroblast growth factor (bFGF) is a potent angiogenic inducer expressed in malignant prostate tissue. In this investigation, serum bFGF levels were measured in men with and without prostate cancer. METHODS: Serum bFGF was measured using a commercial enzyme-linked immunosorbent assay in archival serum from men with various clinical stages of prostate carcinoma. Sera from men with negative systematic sector biopsies and serum prostate specific antigen (PSA) less than 2.0 ng/ml served as controls. Prostate volume was measured with transrectal ultrasound. RESULTS: Serum bFGF level was significantly higher in men with prostate carcinoma compared with those without (P < 0.0007). Among 44 men (11 with carcinoma) with a serum PSA less than 4.0 ng/ml, only bFGF level was associated with carcinoma (P = 0.008). Using a cutoff of 1.0 pg/ml, bFGF afforded a sensitivity of 83% and specificity of 44% in this group. There was no association between bFGF levels and clinical stage, Gleason score, or prostate volume. CONCLUSIONS: These data demonstrate that serum levels of bFGF are elevated in most men with prostatic carcinoma. Although association with stage was not observed, the finding of significant elevation in most men with carcinoma who have "normal" serum PSA levels may indicate diagnostic utility for this analyte.

Topography of neovascularity in human prostate carcinoma.

BACKGROUND: All neoplasms require angiogenesis and resulting neovascularity for growth. The authors and others have confirmed the staging and prognostic significance of quantitative microvascularity density (MVD) in human prostate carcinoma (CAP). In the present investigation, the authors sought to identify the specific site of neovascularity within the neoplasm and adjacent benign tissue. METHODS: Histologically benign and malignant tissues from 14 random radical prostatectomy specimens were studied. The tumor edge was defined precisely by immunohistochemistry, suggesting a high molecular weight cytokeratin that stains only the basal cells of benign histology. Microvascularity density quantification was performed using von Willebrand factor antigen immunohistochemistry as previously defined. Five parallel arcs were defined along which vessel density was calculated including arcs within, on the edge, and removed from the neoplasm. RESULTS: In 13 of 14 cases, the highest vessel density was found within the tumor. Significant differences were observed between the edge of the tumor and 2.5 mm within the benign periphery, between the benign and malignant tissue at the border, and between CAP at the edge and CAP 2.0 mm within the neoplasm. These findings suggest a stepwise increase in MVD toward the center of the neoplasm. CONCLUSIONS: These data confirm the authors' previous observation that prostate cancer has approximately a two-fold increase in MVD compared with the benign tissue. Moreover, high vascularization of the center explains the rare finding of necrosis in CAP. These data suggest that angiogenic promoters may have their highest activity in the center of the neoplasm.

Group B streptococcal infection in children beyond neonatal period: report of 3 patients.

Three patients beyond the neonatal period with group B streptococcal infection were presented, including acute osteomyelitis and septic arthritis, bacteremia and septicemia. Serotyping was done in two patients. One was type Ia and the other was type III. All patients recovered from the disease.

Astrocyte-polymer implants promote regeneration of dorsal root fibers into the adult mammalian spinal cord.

To overcome obstacles to the regeneration of crushed dorsal root fibers at the dorsal root entry zone, we have employed specially designed Millipore implants coated with embryonic astrocytes to serve as a substrate for axonal growth. This strategy was successful in promoting the growth of crushed dorsal root axons into the grey matter of the adult mammalian spinal cord in a small number of animals. Fiber ingrowth into the spinal cord was closely associated with the surface of the polymer implant. In addition, unique terminal arbor malformations, not normally present, were seen in several animals. A consistent finding was the presence of a limited inflammatory response in regions immediately adjacent to the implant where axons penetrate the spinal cord. Our findings suggest that providing the dorsal root entry zone with an embryonic milieu can stimulate a limited amount of axonal regeneration into the adult mammalian spinal cord.

A comparison of the regeneration potential of dorsal root fibers into gray or white matter of the adult rat spinal cord.

To assess the role of white matter inhibition as a barrier to neurite outgrowth in vivo, we unilaterally transected three consecutive lumbar dorsal roots (L4-L6), incised the spinal cord, and transplanted the peripheral stump of L4 either medially onto the white matter of the dorsal columns or laterally, just superficial to the gray matter of the dorsal horn at the level of L5. Three weeks to seven months later, the translocated root was retransected, and its central stump was anterogradely labeled with HRP. The staining pattern demonstrated that regenerating sensory axons had entered the spinal cord from both medially and laterally placed roots. Axonal staining from medially placed dorsal roots (onto the white matter of the dorsal columns) was sparse and limited to the white matter. Staining of laterally placed roots revealed a small subpopulation of regenerating axons which had entered the gray matter and formed terminal arbors. Successful axonal regeneration into the gray matter, albeit minimal, was associated with a localized and limited inflammatory response near the sites of axonal ingrowth.

Surgical removal of cyanoacrylate adhesive after accidental instillation in the anterior chamber.

Cyanoacrylate adhesives are frequently used to seal small corneal perforations. Their use has been limited, however, because of concern over their toxic effect on ocular structures. We describe a post-keratoplasty patient with a wound dehiscence that a surgeon had attempted to seal with cyanoacrylate glue. The glue was accidentally injected into the anterior chamber. After surgical reconstruction of the eye, however, no permanent ocular damage was observed.

Characterization of peripheral blood T lymphocyte subsets in pulmonary sarcoidosis.

The peripheral blood of sarcoidosis patients has been shown to have abnormalities in the T Lymphocyte subsets. There is a consistent reduction in the helper/suppressor T Lymphocyte ratio caused by an increase in suppressor cell percentage as determined by monoclonal antibodies. These findings suggest that the peripheral blood T Lymphocyte subsets may be a useful adjunct for determining the underlying immunological cellular dysfunction in sarcoidosis patients.

Enhanced expression of the c-myc protooncogene in high-grade human prostate cancers.

We examined a series of 29 surgical specimens of benign and malignant human prostate tissue for the expression of both the cHa-ras and c-myc protooncogenes. Northern blots were prepared using polyadenylated mRNA extracted from nine prostatic adenocarcinomas, 19 benign hypertrophied prostates (BPH) and one normal prostate. When the Northern blots were hybridized to a probe for cHa-ras, only one specimen of BPH showed an appreciable amount of the 1.2-kb transcript homologous to cHa-ras. Upon reprobing these blots with c-myc, six cancers showed a considerable amount of a 2.4-kb transcript homologous to c-myc. Three other cancers and all the benign tissue showed little or no detectable 2.4-kb c-myc transcript. On retrospective analysis, the cancers with elevated c-myc transcripts were found to have a Gleason score of 5 and above (poorly differentiated tumors), while the cancers with little or no c-myc transcripts were all of Gleason score 4 and lower. Finally, we compared our ability to detect c-myc transcripts in mRNA extracted from a surgically derived prostate tumor with mRNA extracted from the same tumor subject to a sham electrocautery procedure, as would occur during transurethral resection. The electrocautery procedure decreased both the intensity and the integrity of the c-myc signal in mRNA from the tumor. Thus, our exclusive use of surgically derived prostate tumors may be the reason we are able to detect an elevation in the expression of c-myc mRNA in high-grade tumors.

Hospital-physician relations: overcoming barriers to cooperation.

In the September-October 1986 issue of Physician Executive, we discussed the application of strategic business units (SBUs) to health care. SBUs are those corporate entities that market similar products to one or more target populations with similar characteristics. Examples of SBUs in health care are obstetrics, cardiology, orthopedics, etc. When the services within each SBU are linked together, they might resemble a vertically integrated health care system. In the case of obstetrics, a woman may have contact with physicians, a hospital, home care nurses, house-cleaning services, birthing teachers, and maternity clothing boutiques. Each of these are products/services within the SBU of obstetrics. Strategy development by SBU implies an external focus on the marketplace in terms of the specific mission of the SBU (clinical specialty). It also implies responding to the needs of consumers for whom the historical and present divisiveness between hospitals and physicians is immaterial and irrelevant. In this article, we will focus on ways to stabilize the relationship between hospitals and physicians within an SBU context in order to compete more successfully as a team in today's health care environment.

What business are you in?

Ths structure of the health care delivery system continues to evolve, old organizational types taking new forms and new entities joining in a sometimes helter-skelter array. What is missing is order and purpose. Most health care organizations have not yet asked the critical questions, "What business are we in?" and "Who are our customers?" The authors suggest that what is needed is a strategic business unit approach to the process of business development.

Cations decrease specific [3H]-spiroperidol binding in human prefrontal cortex.

Ligand binding at many physiologically relevant receptors is regulated by divalent cations. To determine whether [3H]-spiroperidol binding sites in prefrontal cortex might be physiologically relevant receptors, we examined the effect of ions on the binding of this ligand in postmortem human prefrontal cortex. Our results indicate that several cations decreased [3H]-spiroperidol binding in a dose-dependent fashion. Of these, Cd++ and Zn++ were the most able to decrease [3H]-spiroperidol binding with IC50 of 5.5 +/- 2.4 X 10(-6)M and 5.6 +/- 1.1 X 10(-5)M respectively. These findings indicate that [3H]-spiroperidol may bind at physiologically relevant receptors in human prefrontal cortex.

Scanning electron microscopy of moist bacterial strike-through of surgical materials.

Scanning electron microscopy was used to demonstrate the process of moist bacterial strike-through of woven and nonwoven surgical materials. Three woven and three nonwoven materials were challenged with an aqueous suspension of Serratia marcescens. The results of these studies confirmed that relatively new, less than 100 cycles of washing and sterilizing, 270 thread Quarpel treated Pima cotton prevents moist bacterial penetration. However, this same woven material when washed and sterilized more than 100 times allowed bacterial penetration. Nonwoven materials prevented penetration only when they were impregnated with plastic or reinforced with a plastic film. Prevention of moist bacterial strike-through of surgical materials, whether they be woven or nonwoven, is dependent upon the effectiveness of their waterproof quality. In woven materials, we have confirmed our previous findings indicating that loss of waterproof characteristics, which occurs after 75 washing-sterilizing cyclings, leads to permeability and to moist bacterial strike-through, regardless of the weave density. In nonwoven materials, dependable resistance to moist bacterial strike, through was achieved only when all moisture penetration was prevented by reinforcement with waterproof plastic film.

Moist bacterial strike-through of surgical materials: confirmatory tests.

New tests consisting of modifications of the inverted Mason jar test confirm our previously reported studies which showed that woven and nonwoven surgical materials vary greatly in their ability to serve as barriers against moist bacterial strike-through. Among the woven materials, only tightly woven Pima cloth or materials treated with Quarpel waterproofing process or with polythene layer lamination was invariably resistant. However, tight-woven Pima cloth, which had been treated with Quarpel became permeable after 100 washing-sterilizing cycles. Of the nonwoven materials, single-layer nonwoven materials tended to unevenly permeable to moist bacterial strike-through. Only the front and sleeves of nonwoven gowns reinforced with polyethelene layer were invariably resistant to moist contamination.