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Biomedical subjects

E Stennert

Publications and source records attributed to E Stennert.

At least 91 records · Page 5Linked to original sources

[Section plastination of the larynx for histology of whole organ sections].

Whole-organ sections of the larynx have evoked the laryngologists' interest for a long time. Established techniques for histological preparation do exist, but these are time-consuming and result in frequent artifacts. Owing to their thicknesses the slices prepared are usually not suitable for a complete histological work-up. The technique of plastination is an established and gentle method for tissue preservation. We describe the technique of sheet plastination to produce whole-organ sections that allow full-scale histological investigation. Deep-frozen larynges are cut into 4-mm-thick sections, dehydrated in acetone and plastinated in epoxy resins. During plastination, fat and lipids are replaced by curable polymer, making decalcification unnecessary. Specimens are cut into thin sections using a diamond-wire saw. An ultra-milling device reduces the section thickness down to about 10 microns. Staining is possible using a variety of special plastination stains, while all routine stains are also applicable after deplastination. Sheet plastination produces artifact- and shrinkage-free whole organ sections in about 1 week. The technique is useful for studies of micromorphometry or tumor spread and can be used to evaluate quickly possible tumor erosion of the laryngeal skeleton after laryngectomy. Since such erosion is one of the criteria for postoperative radiotherapy in many centers, earlier information could make planning easier.

Artifacts↗

Phagocytic microglia during delayed neuronal loss in the facial nucleus of the rat: time course of the neuronofugal migration of brain macrophages.

The injection of Fluoro-Gold (FG) into the whisker pad of rats yields a stable fluorescent labeling of the motoneurons in the lateral facial subnucleus. Following resection of 8-10 mm of the facial nerve, the microglia phagocytose the FG-preloaded neurons and assume the label. Employing this vital labeling of microglia in situ we studied the fate of same after completion of phagocytic activity. Starting at 56 days post resection (DPR) the FG-labeled microglia spread out from the lateral facial subdivision and invaded the entire facial nucleus. The quantitative analysis of this redistribution of the fluorescent marker revealed a prolonged increase in the number of labeled microglia strictly proportional to the delayed loss of neurons. The differentiation between microglia and shrunken neurons was performed with the new method of immunoquenching: the staining of vibratome sections with anti-rat neuron-specific enolase (NSE) combined with an ABC-HRP kit and DAB as detector totally extinguished (quenched) all fluorescence from the pre-labeled facial motoneurons. The fluorescent microglia were additionally stained with GSA I-B4 and OX-42, which should completely quench all fluorescence in the section. However, a few small round cells, always closely opposed to neuronal perikarya, still fluoresced. These NSE-negative, GSA I-B4 and OX-42 negative, but fluorescent cells may represent a new, immunologically uncharacterized microglial cell type, that participates in neuronophagia.

Animals↗

Morphology of experimentally denervated and reinnervated rat facial muscle. I. Histochemical and histological findings.

The morphological changes in rat facial muscles were evaluated after permanent denervation and were compared with findings after immediate reinnervation. Thirty rats underwent transection of the left and right facial nerves immediately followed by hypoglossal-facial nerve anastomosis on the right side (muscular reinnervation) and removal of 8-10 mm of the facial plexus on the left side (permanent muscular denervation). Levator labii muscle samples of both sides were collected sequentially at 2, 6, 7, 10, 20, and 24 weeks after surgery and submitted to routine histological and enzyme histochemical staining procedures. In normal levator labii muscles a typical "chessboard" pattern was found, with type I fibers being smaller than type II fibers. These latter fibers also were more prevalent than the type I fibers. Among the type II fiber subtypes, the type IIB fibers were larger and more frequent. Two weeks after surgery, there were no differences between denervated facial muscles and those undergoing reinnervation. Both showed atrophic myofibers among normal-sized fibers and slight fibrosis. Those muscles denervated for more than 2 weeks displayed increasing fiber atrophy with frequent loss of typability, as well as proliferation of connective tissue and fat cells in perimysial and endomysial sites. After denervation for 20 weeks only a few atrophic fibers were found in wide areas of fibrosis and fat cells. Following nerve anastomosis the reinnervated levator labii muscle showed much less fiber atrophy. Regrowth to normal fiber diameters was found with only a few atrophic myofibers 10 weeks after anastomosis although a moderate fibrosis predominated at perimysial sites.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphatases↗

Differences in glial, synaptic and motoneuron responses in the facial nucleus of the rat brainstem following facial nerve resection and nerve suture reanastomosis.

Transection and reanastomosis of the facial nerve with microsurgical sutures in rats (facial-facial anastomosis) results in the complete regeneration of the facial nucleus, whereas resection of a 10 mm length of the peripheral facial nerve leads to degeneration and loss of neurons in the nucleus. Nerve sutures or resections were performed in 84 female Wistar rats, and the time course and differences between regenerative and degenerative reactions in the facial nuclei were compared after survival times of 4-112 days. The volume of the facial nucleus, number of facial motoneurons and motoneuron density were estimated stereologically by the physical dissector method. Synaptic plasticity, activation of astroglia and microglia were studied cytochemically with anti-synaptophysin, anti-glial fibrillary acidic protein and the isolectin Griffonia simplicifolia I-B4 (GSA I-B4). After facial-facial anastomosis the volume of the facial nucleus and its number of motoneurons remained constant, whereas resection of the facial nerve caused shrinkage of the facial nucleus and loss of one-third of facial motoneurons within 112 days post-operation. Synaptic stripping, activation of microglia and astroglia occurred in the same sequence and were reversible after both operations, but these reactions were more severe and prolonged after resection, i.e. without suture of the facial nerve. It appears to be most important clinically that differences between de- and regeneration become clear within 7 days post-axotomy. Our results strongly support reconstruction of the facial nerve as early as possible after a nerve lesion.

Anastomosis, Surgical↗

Axotomy induces intranuclear immunolocalization of neuron-specific enolase in facial and hypoglossal neurons of the rat.

Neuron-specific enolase as an enzyme of the glycolytic pathway is localized in the cytoplasm of nerve cells, but not in the cell nucleus. We have applied immunocytochemistry with 1:64,000 polyclonal anti-rat neuron-specific enolase to the brainstem of male and female adult Wistar rats following: (a) transection of the facial nerve with immediate microsurgical nerve suture (facial-facial anastomosis), (b) transection of the hypoglossal nerve with immediate suture (hypoglossal-hypoglossal anastomosis) and (c) transection of the facial and hypoglossal nerve with immediate suture of the proximal hypoglossal to the distal facial nerve stump (hypoglossal-facial anastomosis). Studying the intracellular immunolocalization of neuron-specific enolase in neurons of the facial and hypoglossal nucleus we detected that (1) in normal rats about 20% of all facial and hypoglossal neurons display not only cytoplasmic, but also intranuclear neuron-specific enolase-like immunoreactivity and (2) following any axotomy of the facial or hypoglossal peripheral nerve, the perikarya of all injured motoneurons react by an outstanding increase of neuron-specific enolase-like immunoreactivity in the karyoplasm. Similar findings were obtained in experiments on non-fixed cultured Neuro-2a cells that had been lesioned with hydrogen peroxide. Counting the absolute numbers of normal and reactive neurons at 1-365 days post axotomy revealed that the increase of neuron-specific enolase in neuronal cell nuclei is temporary and reversible. It is first detected at 2 days post axotomy, reaches its maximum at 10-18 days post axotomy and is no longer evident 56 days following surgery.(ABSTRACT TRUNCATED AT 250 WORDS)

Anastomosis, Surgical↗

Recovery of original nerve supply after hypoglossal-facial anastomosis causes permanent motor hyperinnervation of the whisker-pad muscles in the rat.

Hypoglossal-facial anastomosis (HFA), used in humans for the treatment of facial palsy, was experimentally performed in adult female Wistar rats. The time course of facial reinnervation and the extent of the new motor nerve supply of the vibrissal muscles that develops after HFA were estimated by counting all motoneurons in the brainstem labeled by injection of horseradish peroxidase (HRP) into the whisker pad; muscle innervation by motor endplates was not studied. In untreated animals, HRP injection labels 1,254 +/- 54 (mean +/- S.D.; n = 6) motoneurons, localized exclusively in the lateral subdivision of the facial nucleus. Immediately following HFA, this number drops to zero. The first HRP-labeled motoneurons appear in the hypoglossal nucleus at 28 days postoperation (dpo) and at 56 dpo their number reaches 1,096 +/- 48. Unexpectedly, the facial nerve, whose proximal stump has been left as blind end during surgery, additionally sends axons to the facial periphery. This resprouting is first detected at 42 dpo with HRP-marked neurons throughout the facial nucleus lacking somatotopic organization. The number of these labeled neurons also rises with time, and at 56 dpo, a total of 1,797 +/- 142 facial and hypoglossal motoneurons, that is, 43% more motoneurons than in normal animals, supplies the whisker pad. This hyperinnervation, that is, the projection of more motoneurons into the target muscle than under normal conditions--further increases to 1,978 +/- 92 motoneurons at 224 dpo and may provide a new animal model for studying the competitive relationships between motoneurons in their search for peripheral targets.

Anastomosis, Surgical↗

Simplified nerve cell counting in the rat brainstem with the physical disector using a drawing-microscope.

A simple modification of the physical disector is presented which is used to count the number of neurons in the hypoglossal nucleus of the rat in a series of paraffin sections. One disector consists of two adjacent sections (6 microm thick) that have been Nissl-stained with cresyl fast violet. In the first step of the procedure each of the two sections in investigated separately with a drawing-microscope. The boundary of the hypoglossal nucleus and the position of neurons devoid of, or containing a part of, the cell nucleus in the plane of the section are marked on transparent paper. In the second step, these two drawings are placed one upon another, aligned and the number of cell profiles that show a cell nucleus in one but not in both drawings counted. This modification of the disector method for cell counting needs no specialized equipment, simply a light microscope with drawing apparatus, and can be combined with histochemical studies of other sections from the same tissue block.

Animals↗

The hypoglossal-facial anastomosis as model of neuronal plasticity in the rat.

Hypoglossal-facial cross anastomosis (HFA) causes regeneration with change of function, as the axotomized hypoglossal motoneurons sprout into the facial plexus and reinnervate the mimic musculature. Following HFA, hypoglossal-hypoglossal single anastomosis (HHA) and resection of 8-10 mm peripheral hypoglossal nerve in 190 female adult Wistar rats, we compared the axon reactions in the hypoglossal nucleus during 1) regeneration with change of function, 2) regeneration with restoration of original function and 3) degeneration of the nucleus. Following postoperative survival times of 1-16 weeks we estimated the volume of the hypoglossal nucleus and counted the number of hypoglossal neurons with the physical disector on both sides of the brainstem. Additional sections of the same animals were reacted with anti-synaptophysin, anti-GFAP and the isolectin Griffonia simplicifolia I-B4 (GSA I-B4) as cytochemical markers for presynaptic boutons, activated astroglia and microglia. After HHA and HFA all hypoglossal neurons survive and the volume of the hypoglossal nucleus remains constant. Resection of the hypoglossal nerve leads to the loss of one third of the hypoglossal neurons and of one third of the volume of the hypoglossal nucleus within 16 weeks post operation. Hypoglossal-facial anastomosis and hypoglossal-hypoglossal anastomosis differ in postoperative swelling of the hypoglossal nucleus, microglia and astroglia activation and the duration of synaptic stripping. All differences are limited to the acute growth phase during regeneration. It is concluded that hypoglossal-facial anastomosis provides more stimulation and facilitates faster recovery of the hypoglossal nucleus than does hypoglossal-hypoglossal anastomosis.

Anastomosis, Surgical↗

[Endoscopically-controlled endonasal orbital decompression in malignant exophthalmos].

In 6 patients with endocrine ophthalmopathy, indications, surgical technique and results of the endoscopic controlled endonasal orbital decompression are described in comparison to the common surgical procedures. When medical and radiation therapy fail, indications for decompression are a) loss of visual acuity or visual field defects, b) increasing strabismus, c) severe keratopathy due to eyelid retraction. The endoscopic-controlled endonasal surgical decompression technique is proceeded in three steps. First, an endonasal ethmoidectomy with resection of the middle turbinate is performed and the medial wall of the maxillary sinus is widely opened. Second, the medial and inferior wall of the orbital walls are removed, preserving the infraorbital nerve. In the last step, the periorbital area is incised and the orbital fat herniates. The advantages of this procedure consist in the absence of exterior scars and the known morbidity of a Caldwell-Luc antrotomy. The results were documented by computed tomographic scans (CT), magnetic resonance imaging (MRI), Hertel measurements, evaluation of ocular motility and ophthalmoscopy. An average of 3-4 mm improvement in Hertel-measurements could be reached. All patients had a postoperative improvement of visual acuity. 2 patients developed more significant diplopia postoperatively, whereas in all other patients ocular motility either improved or rested unaffected. Therefore, the endoscopic controlled endonasal procedure allows to obtain comparable results to the common extranasal and transantral procedures without the disadvantages of the latter.

Aged↗

Carboplatin. The better platinum in head and neck cancer?

Chemotherapeutic regimens containing cisplatin are the most effective ones in the treatment of squamous cell carcinoma of the head and neck. Because of the high rate of dose-limiting side effects of cisplatin, carboplatin, a second-generation cisplatin analogue, was tested in a phase II trial with fluorouracil in 55 previously untreated patients with advanced carcinoma of the head and neck. Among the 52 patients who completed the study, there were 17 complete responses (33%), 28 partial responses (54%), five patients with no change (10%), and two with progressive disease (4%). Toxic side effects of all courses summed together included leukopenia in 65% of courses, thrombocytopenia in 45% of courses, nausea or vomiting in 29% of courses, and change in serum creatinine level in 3% of courses. These data were compared with the results of our study with cisplatin and fluorouracil in comparable patients and indicated that carboplatin and fluorouracil is better for induction chemotherapy in the treatment of head and neck cancer than cisplatin and fluorouracil due to similar effectiveness but less toxic effect.

Antineoplastic Agents↗