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Biomedical subjects

M L Bates

Publications and source records attributed to M L Bates.

18 recordsLinked to original sources

Inhibitory proteoglycan immunoreactivity is higher at the caudal than the rostral Schwann cell graft-transected spinal cord interface.

To begin to evaluate the influence that proteoglycans may have on the success of Schwann cell (SC) transplants to induce axonal regrowth across a complete transection lesion and beyond, we determined the pattern of expression of inhibitory chondroitin sulfate proteoglycans (CSPGs) 3 weeks after transplantation into completely transected adult rat thoracic spinal cord. Using immunohistochemistry, we observed that: (1) CSPGs recognized by CS-56 antibody are present on astrocytes, fibroblasts, and SCs in the distal graft, and at lesion and cystic cavity borders; (2) CS-56 immunoreactivity (IR) is greater at the caudal SC graft-host cord interface than the rostral interface; (3) phosphacan-IR, also greater at the caudal interface, is associated with astrocytes, fibroblasts, as yet unidentified cells, and extracellular matrix; (4) neurocan-IR is present on astrocytes and as yet unidentified cells in grey and white matter; and (5) NG2-IR is associated with matrix near SC grafts, unidentified cells mainly in white matter, and lesion borders and cysts. Neither oligodendrocytes nor activated macrophages/microglia were immunostained. In sum, the CSPGs studied are increased at 3 weeks, especially at the caudal SC graft-cord interface, possibly contributing to an inhibitory molecular barrier that precludes regrowing descending axons from entering the caudal host cord.

Animals↗

Axonal regeneration into Schwann cell grafts within resorbable poly(alpha-hydroxyacid) guidance channels in the adult rat spinal cord.

Axonal growth and myelination in a SC graft contained in a resorbable tubular scaffold made of poly(D,L-lactic acid) (PLA50) or high molecular weight poly(L-lactic acid) mixed with 10% poly(L-lactic acid) oligomers (PLA(100/10)) were studied for up to 4 months after implantation in the completely transected adult rat thoracic spinal cord. The PLA50 tubes collapsed soon after implantation and, consequently, compressed the graft inside, leading to only occasional thin cables with SCs and a low number of myelinated axons: 17 +/- 6 at 1 and 158 +/- 11 at 2 months post-grafting. The cable contained 32 +/- 23 blood vessels at 2 weeks, 55 +/- 33 at 1 month and 46 +/- 30 at 2 months after implantation. PLA(100/10) tubes, on the other hand, were found to break up into large pieces, which compressed and sometimes protruded into the tissue cable inside. At all time points studied, however, cables contained SCs and were well vascularized with 414 +/- 47 blood vessels at 2 weeks, 437 +/- 139 at 1, 609 +/- 134 at 2 and 396 +/- 95 at 4 months post-grafting. The number of myelinated axons was 712 +/- 509 at 1 month, 1819 +/- 837 at 2 months and 609 +/- 132 at 4 months post implantation. These results demonstrated that fiber growth and myelination into a SC graft contained in a resorbable PLA(100/10) tube increases over the first 2 months post-implantation but decreases thereafter. Changes in geometry of both types of polymer tubes were detrimental to axonal regeneration. Future research should explore the use of polymers that better retain the appropriate mechanical, geometrical and permeability properties over time.

Animals↗

Microleakage and adaptation of Class II packable resin-based composites using incremental or bulk filling techniques.

PURPOSE: To compare the ability of three packable resin-based composite (RBC) systems (ALERT, SureFil, and Solitaire), placed and light-cured in either incremental layers or in bulk, to seal the gingival margin of Class II preparations when the gingival margin is placed apical to the cementoenamel junction. MATERIALS AND METHODS: Packable RBCs were used to restore 60 extracted human premolars with Class II preparations using an incremental-cure technique (each layer no greater than 2 mm in thickness when light-cured) or a bulk-cure technique (n=10). Manufacturers' directions were followed; the cure technique was the only variable. After the restorations were completed, the specimens were thermocycled, stained, sectioned, and viewed under a light microscope for microleakage at the gingival margin. RESULTS: All specimens showed leakage at the gingival margin. No statistically significant difference was found between the bulk-cure and incremental-cure techniques for each of the RBC systems evaluated. ALERT had significantly less leakage than SureFil and Solitaire.

Acid Etching, Dental↗

Dental treatment guidelines for use of restraints within the nine Louisiana developmental centers. Louisiana State University Dental Health Resources Program.

There are approximatively 2000 mentally retarded or developmentally disabled (MR/DD) individuals in Louisiana's nine developmental centers (LDC). Two-thirds resist the delivery of preventive and clinical dental services. Therefore, for these patients to receive quality, comprehensive dental care, some form of restraint is necessary. Physical holds, mechanical and/or chemical restraints allow caregivers and dental professionals to provide medical/dental services. However, prior to the use of restraint, a clear understanding and acquisition of informed consent are needed. The dentist, developmental center and its administration are responsible for developing a plan to obtain informed consent and to express the importance of restraint to each resident's guardian. This article defines the types of restraint based upon resistance encountered and defines informed consent according to the Louisiana Medical Consent Law.

Dental Care for Persons with Disabilities↗

Characterization of photochemically induced spinal cord injury in the rat by light and electron microscopy.

This study characterized by light and electron microscopy 49 photochemically induced lesions in adult rat spinal cord at 16 time intervals from 2 days to 17 months after lesioning. Vascular thrombosis, resulting from an intravascular photochemical reaction induced by a rose bengal/laser beam interaction, led within a few days to an extensive area of tissue deterioration. This area, termed the "lesion cavity" in contrast to the "secondary cavity" observed later, was at least 6 mm long and, at the epicenter, extended across most of the spinal cord width and from the dorsal surface to a level near the central canal. The area of spared tissue, 43% of the spinal cord cross-section at 2 days, did not change significantly between 2 and 56 days. Large numbers of macrophages populated the degenerating area by 5 days. This necrotic area was surrounded by a thin peripheral rim of largely intact white matter dorsally and laterally except at the epicenter where the white matter degenerated dorsomedially. In these peripheral regions, demyelination and, by 14 days, remyelination by both oligodendrocytes and Schwann cells (SCs) were evident. By 28 days, far more SCs (and meningeal cells) had entered the dorsal spinal cord, typically at the epicenter where meningeal thickening was most striking, and had migrated farther into the lesion cavity. These SCs and the axons they myelinated remained prominent in dorsal regions for many months, particularly at the epicenter; the proportion of SC to oligodendrocyte myelin diminished away from the epicenter. By 8 weeks, the lesion cavity was considerably diminished in size and thereafter it contained scattered macrophages, SC-myelinated axons, and blood vessels, primarily medially owing to flattening into clefts bilaterally. The cavity was partly bordered by astrocytes whose surfaces toward the lesion cavity were highly irregular and coated with basal lamina. Bare axons, consistently seen by electron microscopy at 5 days to 6 months, were typically ensconced among astrocytes starting at 28 days. Also by this time large, smoothly contoured, empty secondary cavities appeared, usually rostral and caudal to the epicenter; they did not increase in size or number with time. From 28 days to 17 months postlesion they occurred in 68% of the lesioned spinal cords. The secondary cavity border was composed of cells thought to be astrocytes but, surprisingly, the luminal surface was smooth and lacked basal lamina, in contrast to the primary lesion cavity border. Thus, two types of cavities formed after photochemical lesioning. This lesioning technique may provide an appropriate milieu to better understand aspects of the vexing problem of post-traumatic syringomyelia in the human.

Animals↗

Regrowth of axons in lesioned adult rat spinal cord: promotion by implants of cultured Schwann cells.

Highly purified populations of Schwann cells were grafted into lesioned adult rat spinal cord to determine if they promote axonal regeneration. Dorsal spinal cord lesions were created by a photochemical lesioning technique. Schwann cells derived from E16 rat dorsal root ganglia, either elongated and associated with their extracellular matrix or dissociated and without matrix, were rolled in polymerized collagen to form an implant 4-6 mm long which was grafted at 5 or 28 days after lesioning. No immunosuppression was used. Acellular collagen rolls served as controls. At 14, 28 and 90 days and 4 and 6 months after grafting, animals were analysed histologically with silver and Toluidine Blue stains and EM. The grafts often filled the lesion and the host borders they apposed exhibited only limited astrogliosis. By 14 days, bundles of unmyelinated and occasional thinly myelinated axons populated the periphery of Schwann cell implants. By 28 days and thereafter, numerous unmyelinated and myelinated axons were present in most grafts. Silver staining revealed sprouted axons at the implant border at 28 days and long bundles of axons within the implant at 90 days. Photographs of entire 1 micron plastic cross-sections of nine grafted areas were assembled into montages to count the number of myelinated axons at the graft midpoint; the number of myelinated axons ranged from 517-3214. Electron microscopy of implants showed typical Schwann cell ensheathment and myelination, increased myelin thickness by 90 days, and a preponderance of unmyelinated over myelinated axons. Random EM sampling of five Schwann cell grafts showed that the ratio of unmyelinated to myelinated axons was highest (20:1) at 28 days. These ratios implied that axons numbered in the thousands at the graft midpoint. Dissociated Schwann cells without matrix promoted axonal ingrowth and longitudinal orientation as effectively as did elongated Schwann cells accompanied by matrix. There was a suggestion that axonal ingrowth was at least as successful, if not more so, when the delay between lesioning and grafting was 28 rather than 5 days. Acellular collagen grafts did not contain axons at 28 days, the only interval assessed. In sum, grafts of Schwann cells in a rolled collagen layer filled the lesion and were well tolerated by the host. The Schwann cells stimulated rapid and abundant growth of axons into grafts and they ensheathed and myelinated these axons in the normal manner.

Animals↗

An animal model for pharyngocutaneous fistulas.

Postoperative pharyngocutaneous fistula is not an uncommon complication. Although the frequency of postoperative fistulae has decreased with the use of perioperative broad-spectrum antibiotics, it remains a complication with significant morbidity and expense. We present an animal model for postoperative pharyngocutaneous fistulae based on increasing wound tension. The New Zealand white rabbit was used to assess the rate of wound breakdown in the thyrohyoid membrane. The animals were assigned to one of seven groups according to the width of tissue resected. After tissue resection, the pharyngeal wounds were repaired, as were the overlying skin wounds. Animals were monitored postoperatively up to 14 days, at which time they were killed and underwent autopsy. Statistically significant results were achieved that demonstrate an increasing incidence of pharyngeal wound breakdown associated with increasing width of tissue resected and, therefore, closure tension. The procedure and results will be presented in detail. We propose that this model may be used to assess postoperative wounds as well as substances or methods touted as promoters of wound healing.

Animals↗

Perineurium originates from fibroblasts: demonstration in vitro with a retroviral marker.

A cellular sheath, the perineurium, forms a protective barrier around fascicles of nerve fibers throughout the peripheral nervous system. In a study to determine the cellular origin of perineurium, a culture system was used in which perineurium forms after purified populations of sensory neurons, Schwann cells, and fibroblasts are recombined. Before recombination, the Schwann cells or the fibroblasts were labeled by infection with a defective recombinant retrovirus whose gene product, beta-galactosidase, is histochemically detectable in the progeny of infected cells. Perineurial cells were labeled when fibroblasts had been infected but not when Schwann cells had been infected. Thus, perineurium arises from fibroblasts in vitro and, by implication, in vivo as well.

Animals↗

Light microscopic immunocytochemical localization of hepatic and intestinal types of fatty acid-binding proteins in rat small intestine.

Monospecific antisera to purified hepatic fatty acid-binding protein (hFABP) and gut fatty acid-binding protein (gFABP) have been used to localize these two proteins in the small intestine of fed rats at the light microscopic level. Pieces of duodenum, jejunum, and ileum were removed from 4-, 10-, 20-, 22-, and 60-day-old Sprague-Dawley rats. Both cryostat and paraffin sections were studied for the presence of hFABP or gFABP by the avidin-biotin immunoperoxidase method. Slides were graded blind for the intensity of staining. Despite the structural and immunological differences between these two proteins, we showed no major differences between their staining patterns or their staining intensity throughout the intestine during postnatal development. The staining for both fatty acid-binding proteins was cytoplasmic. No brush border staining was found. Staining was more intense in the proximal rather than distal intestine, in the villus rather than crypt cells, and in the apex rather than the base of intestinal cells. Shifts in staining patterns, and staining intensity occurring during development may be related to variations in dietary fat intake, rates of cell proliferation, intestinal anatomy, and mechanisms for fat absorption.

Animals↗

Scanning electron microscopic appearance of chronic ulcerative colitis with and without dysplasia.

This study was conducted to determine whether scanning electron microscopy of colonic mucosal biopsy specimens can help to detect dysplasia in patients with chronic ulcerative colitis. In the first phase of the study, using light microscopy as the standard for the diagnosis, the scanning electron microscopic appearance of specimens from patients with chronic ulcerative colitis and control patients was examined. Descriptive criteria were established to identify normal, atrophic, and dysplastic colonic mucosa. In the second phase, quantitative techniques were used to develop more objective criteria for the diagnosis of dysplasia in ulcerative colitis. Twenty-one coded colonic specimens from 11 patients were sequentially examined by scanning electron microscopy and by light microscopy. The three morphometric analyses performed on the surface epithelial cells were number of cells per unit area, number of microvilli per unit area, and percentage of microvilli with a normal width. The cell count and percentage of microvilli with a normal width were significantly reduced in the seven specimens with colonic dysplasia as compared with non-dysplastic tissues. Scanning electron microscopy may serve as an adjunct to light microscopy in the diagnosis of colonic dysplasia.

Atrophy↗

Distribution of immunoreactive alkaline phosphatase in the adult rat ileum by immunoperoxidase staining at the light microscopic level.

Our prior immunocytochemical studies using monospecific antibody to alkaline phosphatase, Bouin's fixation, and paraffin sections demonstrated a decreasing gradient of villus brush border staining from the proximal to the distal rat small intestine. In addition, we noted an unusual pattern of staining in the terminal centimeter of the adult rat ileum: the villus brush border staining was less intense than crypt brush border staining. To determine whether this pattern of staining was present throughout the entire ileum, we examined alkaline phosphatase staining in two separate jejunal sites and the entire lowest third of the intestine of adult Wistar rats. With Bouin's fixation and paraffin embedding, both conventional and germ-free rats showed the same unusual staining pattern throughout the entire ileum. This pattern suggested that bacterial proteases were not responsible for the diminished ileal brush border alkaline phosphatase. However, when acetone fixation and cryostat sections were used with the avidin-biotin-peroxidase complex system, the previously noted reversed gradient of staining between the ileal villus and crypt areas was no longer present. Rather, ileal crypt brush border staining was less than ileal villus brush border staining. With either methodology, jejunal villus brush border staining was significantly more intense than ileal brush border staining, whereas the deep crypt brush border staining was not significantly different between the two regions. The present study reinforces the need for a combination of methodologies in order to best and most accurately localize certain antigens with immunocytochemistry. It also confirms a decreasing proximal to distal gradient for villus brush border alkaline phosphatase despite similar deep crypt brush border staining throughout the small intestine.

Alkaline Phosphatase↗

Esophageal dysplasia. Assessment by light microscopy and scanning electron microscopy.

This study was performed to determine the characteristics of esophageal dysplasia by scanning electron microscopy. A total of 82 esophageal biopsy specimens were taken from 30 patients who were divided into three groups. Group 1 patients had no known esophageal disease. Group 2 patients had squamous cell cancer. Group 3 patients had esophagitis. Mucosal biopsy specimens that had been diagnosed by light microscopy as normal, esophagitis, or dysplastic mucosa were examined by scanning electron microscopy. A characteristic appearance for each type of mucosa was recognized by scanning electron microscopy. A quantitative analysis of the scanning electron microscopy feature of microridge density showed a statistically significant difference not only between normal and dysplastic mucosa, but also between esophagitis and dysplastic mucosa. The addition of scanning electron microscopy to light microscopy may prove helpful in the diagnosis of dysplasia as well as in the understanding of the biologic behavior of dysplastic cells and possibly their relationship to esophageal carcinoma.

Adult↗

A note on inhibition test and electrophoretic detection limits of antibiotics used in British animal husbandry.

A four plate microbiological inhibition test (the FPT) and a bioelectrophoretic method were evaluated for their ability to detect a range of antibiotic agents, which may be present as residues in animal tissues following their therapeutic use in animal husbandry. Both methods exhibited a wide range of sensitivities and several of the tested antibiotics could not be detected by either method. The pattern of responses across the bacterial plates in the FPT could not be used to identify agents and the bioelectrophoretic inhibition zone diameters were generally too large to allow the use of Rs values for identification. The Bacillus subtilis pH 7.2 plate with trimethoprim added was as effective as the four bacterial plates used in the FPT in antibiotic detection.

Animals↗

Localization of immunoreactive alkaline phosphatase in the rat small intestine at the light microscopic level by immunocytochemistry.

Monospecific antibody to purified alkaline phosphatase hs been used to localize alkaline phosphatase in the rat small intestine at the light microscopic level. Pieces of duodenum, jejunum, and ileum were removed from 2-, 9-, 12-, 18-, 21-, 26-day-old and adult wistar rats. They were fixed in Bouin's fluid and examined for the presence of alkaline phosphatase by th immunoperoxidase method. Slides were graded blindly for the intensity of staining. The localization of alkaline phosphatase by the immunoperoxidase method extends previous histochemical observations in several ways. First, diffuse cytoplasmic staining is present particularly in the opical portion of the villus and crypt epithelium. Second, staining for alkaline phosphatase is present on the brush border and in the apical portion of the deep crypt cells throughout the duodenum, jejunum, and ileum at the various ages tested. Third, in the adult rat distal ileum there is more staining on the brush border of the deep crypt epithelial cells than on the villus absorptive cells. These observations are consistent with the presence of a non-brush border alkaline phosphatase in all intestinal cells and with fan enzymatically inactive form of alkaline phosphatase in the deep crypt epithelium.

Alkaline Phosphatase↗

Radiological/pathological correlations in uremic bone disease.

Skeletal radiographs and non-decalcified bone specimens from 17 chronically uremic patients with radiographic evidence of bone disease were studied quantitatively. The results of each morphological technique were compared in an attempt to define the roentgenographic manifestations of renal osteodystrophy histologically. The radiographs correlated best with the trabecular bone manifestations of osteitis fibrosa but showed poor correlation with histological evidence of osteomalacia. Radiographic signs of osteosclerosis could not be correlated with any radiographic or histiolgical feature of bone resorption.

Adult↗

The effects of anesthetics on an in-vivo oxygen electrode.

The IBC in-vivo oxygen electrode may be used during anesthesia. The possibility that three common inhalation anesthetics--halothane, enflurane, and methoxyflurane--might be reduced at the electrode, thereby changing the PO2 value, was tested. Enflurane and methoxyflurane do not affect the current produced at the electrode; halothane significantly increases PO2 readings at oxygen tensions below 105 torr. It is suggested that PaO2 readings below 105 torr be confirmed by the standard bench method.

Anesthetics↗

Determination of synthetic growth promoters in bile.

To monitor the use of synthetic growth promoting hormones in red meat animals requires a sensitive and rapid method of analysis. This paper describes the development of such a method for these agents in bile, based on a high pressure liquid chromatographic purification procedure with detection and estimation by radioimmunoassay.

Anabolic Agents↗