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PubMed1999

PCNA bearing structures are retained in apoptotic phase of childhood ALL cell cycle.

BACKGROUND: Drug resistance to DNA directed therapy may depend on proliferative as well as apoptotic cell fraction. PCNA/Ki67 ratio excess, possibly reflecting DNA excision repair, is of additional interest to drug resistance in MTT testing. The cell cycle phase/antigen expression pattern in childhood acute lymphoblastic leukemia (ALL) is not known. AIMS: To study the relationship between nuclear expression of PCNA, Ki-67 and Frag-EL positivity in childhood ALL. METHODS: 1.3.1. Study Groups. Diagnostic bone marrow trephine biopsies of 32 consecutive unselected cases of childhood ALL were included in the study. 1.3.2. Immunohistochemistry. Commercially available Moab PCNA (PC10, DAKO, USA), Ki-67 (MM1, NovaCastra, UK) were used to label cycling cells in routinely processed 5 microns paraffin sections. 1.3.3. In-Situ Labelling of Apoptotic Cells. The 3'-OH ends of apoptosis specific DNA fragments were labelled in-situ on subsequent 10 microns sections (Frag-EL, CalBiochem, USA). 1.3.4. Quantitation. After blinding and randomisation, 10 systematic random fields of > 20 nuclei and nuclear size bias correction was used to determine positive nuclei fraction. RESULTS: While the sum of apoptotic and proliferative cell fraction (Ki-67 + Frag-EL%) equalled 100% in 5/32 cases, PCNA expression into at least the early phases of apoptosis ([%PCNA-%K-67] > [100-%Frag-EL] was found in 17/32 cases. CONCLUSIONS: PCNA/Ki67 ratio excess may not reflect DNA excision repair activity but rather slow degradation of antigen bearing structures limiting relevance to drug resistance study.

Apoptosis↗
PubMed2003

Correlation of lesion volume and brain swelling from a focal brain trauma.

Brain edema and secondary growth of a traumatic brain tissue necrosis are important manifestations of secondary brain damage and of prognostic significance in severe head injury. Aim of the current study was to analyze the interdependency of the resulting brain swelling from the size of the focal traumatic lesion. Male Sprague-Dawley rats were intubated and mechanically ventilated. A trephination was made over the left parietal cortex for induction of a cold lesion. Different injury severities were achieved by varying the contact time of the cooled copper-cylinder and the exposed cortex. Animals were randomized into 12 experimental groups. Hemispheric brain swelling was measured in groups A1-A6 (n = 4-8) by gravimetry 24 hrs after lesions of six increasing severity levels. Correspondingly, in animals of groups B1-B6 (n = 5-7) the volume of necrosis was planimetrically assessed in histological serial sections of the brain obtained 24 hrs after trauma of different severity. In groups A1-A6. hemispheric brain swelling (increase in weight) was growing with increasing contact duration of the cold probe with the exposed cerebral cortex, i.e. from 7.7 +/- 0.4% (5 s) to a maximum of 9.9 +/- 0.5% (25 s). Longer contact periodes (30 s) were not further effective to increase hemispheric brain swelling. The contact times and extent of swelling were linearly correlated between 5 s and 25 s (r = 0.47; p < 0.01). The volume of necrosis in groups B1-B6 increased from 35.7 +/- 3.7 mm3 (5 s) to 106.3 +/- 10.3 mm3 (30 s). There was again a linear correlation between the duration of contact of the cold probe (i.e. injury severity) with the brain cortex and the volume of necrosis (r = 0.77; p < 0.01). Accordingly. the lesion volume could be increased in a reproducible manner from 35.7 up to 106.3 mm3 by extending the contact times of the cooling device and cerebral cortex. Hemispheric swelling, predominantly due to vasogenic brain edema, was expanding in relationship with the volume of necrosis.

Animals↗
PubMed2000

LF16-0687 a novel non-peptide bradykinin B2 receptor antagonist reduces vasogenic brain edema from a focal lesion in rats.

Head injury world wide is still the most frequent cause of morbidity and mortality among the population under 45 years. Approximately 50% of patients dying from severe head injury have a therapy refractory intracranial pressure rise (Baethmann 1998). Traumatic brain edema, e.g. resulting from disruption of the blood-brain barrier is viewed as an important factor of the increased intracranial pressure. Bradykinin, an active peptide of the kallikrein-kinin system is considered to enhance brain edema formation which is attributed to its permeabilizing effect on the blood-brain barrier and on dilation of arterial blood vessels in the brain mediated by B2-receptors facilitating extravasation. Currently, LF16-0687, a novel non-peptide bradykinin B2 receptor antagonist was experimentally tested as to its therapeutical potential on vasogenic brain edema from a cortical focal lesion. Following trephination of the skull in anaesthesia, male Sprague-Dawley rats were subjected to a focal cold injury of the left parietal cortex. Animals of two experimental groups were receiving either LF16-0687 as high or low dose, whereas one group of untreated animals with trauma was treated with 0.9% NaCl as continuous infusion beginning 10 min before until 24 h after lesion. 24 h after trauma the brain was removed from the skull, and the cerebral hemispheres were separated in the median plane for gravimetric assessment of hemispheric swelling. No significant reduction of hemispheric brain swelling (+7.4 +/- 2.9%) was found in animals receiving high-dose LF16-0687 as compared to the untreated controls. Brain swelling, however was significantly attenuated by the low-dose treatment, i.e. to +6.4 +/- 1.3%; vs. +9.3 +/- 1.1% found in the controls, (p < 0.05). The current data confirm that blocking of bradykinin B2-receptors by LF16-0687 is significantly attenuating vasogenic brain edema from a focal cold lesion. The therapeutical properties of the antagonist on brain edema formation cannot be attributed to a lowering of the blood pressure. Rather, specific blocking effects of B2-receptors in the brain appear to be involved. In conclusion, the understanding of secondary brain damage including brain edema in head injury has been markedly enhanced by the discovery of pathophysiologically active mediator compounds playing a role in its various manifestations. The current data confirm a pathophysiological function of bradykinin in vasogenic brain edema mediated by activation of B2-receptors. Currently it is studied whether LF16-0687 also reduces brain swelling when given after an insult.

Animals↗
PubMed1994

Endoscopic surgery of traumatic intracranial haemorrhages.

The results of endofiberscopic removal of traumatic intracranial haematomas and hygromas in 180 patients are analysed. Peculiarities of the surgical techniques using flexiscopes and original devices in epidural, subdural, intracerebral, intraventricular haemorrhages of various consistencies, size and location are reported. A technique of the trephination access and delayed cranioplasty for endoscopic removal of extensive subdural and intracerebral haematomas is presented. Indications, contra-indications for endoscopic haematoma surgery, advantages, disadvantages, failures of the techniques are discussed.

Brain Injuries↗
PubMed1980

Combined approach (stereotactic-microsurgical) to a paraventricular arteriovenous malformation. Case report.

The authors present one case of a 42 years old male patient who had a small right paraventricular arteriovenous malformation. At operation the patient was placed in the Talairach's stereotactic frame and, after performing a right carotid angiogram to locate the lesion exactly, a flexible cannula was inserted stereotactically, through a previous trephine hole, until the tip reached the AVM. Using a microsurgical technique and through a 1-2 cm corticotomy the cannula pathway was followed easily reaching the AVM. Total removal of the AVM was confirmed by a carotid angiogram, the patient was discharged without symptoms 7 days later. The authors propose that due to the special design of the Talairach's stereotactic frame, its application as a locating system for small and deep AVM's would greatly facilitate their removal using microsurgical techniques.

Adult↗
PubMed1994

Therapeutical efficacy of a novel chloride transport blocker and an IP3-analogue in vasogenic brain edema.

The efficacy of torasemide, a novel chloride-channel blocker, and of PP56, an IP3 analogue, was currently examined in experimental brain edema. Following trephination in anesthesia rats were subjected to a focal cold injury of the left cerebral hemisphere. Animals of 4 experimental groups receiving either torasemide (i.v. at 30 min before and 6 h after lesion) or PP56 (continuous infusion beginning at 30 min before until 24 h after lesion) at two dose levels were compared with controls administered with i.v. saline. 24 h after trauma the brain was removed from the skull, and the hemispheres were separated in the median plane for gravimetric assessment of hemispheric swelling. Hct, blood gases and body temperature remained constant in all groups. Blood pressure was found to increase in a dose-dependent manner in animals with torasemide. No significant reduction of brain swelling was found in animals with low-dose torasemide (8.51 +/- 0.63%) or low- (7.91 +/- 0.60) and high-dose PP56 (6.85 +/- 1.05%) as compared to the untreated controls. Brain swelling, however, was significantly attenuated by high-dose torasemide to 7.04 +/- 0.36%, as compared to 8.89 +/- 0.29% of the untreated group (p < 0.005). It is currently studied whether torasemide reduces brain swelling when given after the insult.

Animals↗
PubMed1982

Ketoconazole: a new imidazole antifungal agent has both prophylactic potential and therapeutic efficacy in keratomycosis of rabbits.

A highly reproducible quantitative model of fungal infection of the rabbit's corneal stromal was produced using multiple corneal microtrephination. Aspergillus flavus (K4/77), at a concentration of 10(6) spores per ml was systematically implanted into the trephine sites in the cornea, and the degree of corneal infection determined. By pre-inoculation and post-inoculation challenge of these cornea with 1% ketoconazole in arachis oil, the prophylactic potential and the therapeutic usefulness of ketoconazole was determined. Ketoconazole, acetyl-dichlorophenyl-imidazole, has a significant prophylactic potential in inhibiting the development of corneal stromal fungal lesions when it is administered to the cornea of New Zealand white male rabbits as a 1% solution in arachis oil for two consecutive times hourly for two hours before the inoculation of the rabbits cornea with an ocular pathogenic Aspergillus flavus. Ketoconazole also has a therapeutic effect in the reduction of well established A. flavus keratitis in rabbits. When administered as 1% solution in arachis oil for ten consecutive hours daily to well established A. flavus lesions of the cornea of New Zealand albino rabbits, ketoconazole took about sixteen days to cure all the corneal lesions. Finally, using a yeast nitrogen base liquid medium, the in vitro minimal inhibitory concentrations of ketoconazole to twenty-five various human ocular pathogenic fungal isolates were determined and used to recommend those fungi for which ketoconazole would be a good choice for therapy.

Animals↗
PubMed1992

Lamellar excimer laser keratoplasty: reproducible photoablation of corneal tissue. A laboratory study.

We examined the depth of ablation of the recipient bed with different counts of oscillations of excimer laser beam, to determine the correlation between planned and real depth. The ablation rate per oscillation was tested preoperatively by blackened photographic paper of defined thickness and thus was calculated to be 5 microns. Forty pig eyes were used for the first study. Each eight eyes were ablated in the planned depth 100 microns, 200 microns, 300 microns, 400 microns and 500 microns. The corneal thickness was measured with an ultrasonic pachymeter before and after the procedure. The depth measured after the photoablation was 99.4 +/- 36.4 microns for 100 microns planned depth, 186.7 +/- 55.3 microns for 200 microns, 298.4 +/- 68.5 microns for 300 microns, 373.9 +/- 65.7 microns for 400 microns and 480.1 +/- 59.3 microns for 500 microns. Comparing the depth measured after the photoablation to planned depth, there was a significant correlation (correlation coefficient: R = 0.93; p < 0.0001). Five other corneas trephinated from pig cadaver eyes were ablated from the endothelial side to the desired thickness (100 to 500 microns) of lamellar graft. In a second step a donor mask was placed onto the cornea and a laser light spot was led until perforating on all sides. The lamellar keratoplasty was completed by suturing the corneal graft into the bed. Macroscopic and microscopic examination of sutured eyes after fixation showed a good fit of wound margins and stromal interface. These results indicate that excimer laser is useful for reproducible corneal photoablation in lamellar keratoplasty.

Animals↗
PubMed1994

Megakaryocytes and sinus walls in primary osteomyelofibrosis: transendothelial migration as revealed by three-dimensional reconstruction of serial sections following sequential double-immunostaining.

Using sequential double-immunostaining and a newly-developed three-dimensional (3D-) reconstruction technique on serially cut sections from bone marrow trephines, we studied the transmural passage of megakaryocytes through the sinus wall. Biopsies derived from patients with primary (idiopathic) osteomyelofibrosis were exposed to monoclonal antibody against type IV collagen to delineate the sinus walls and also the frequently thickened basement membrane. Staining with the primary antibody was followed by Y2/51 (CD61) to identify all elements of megakaryopoiesis. In most instances serial sectioning and 3D-reconstruction revealed an amoeboid shape of megakaryocytes and a tandem-like arrangement in close spatial contact with the abluminal surface of the sinus wall. Preceded by formation of cytoplasmic processes, straight penetration of entire megakaryocytes through gaps in the sinus walls into the lumen was seen. Where collagen deposits apparently presented a barrier, a mole-like tunnelling through the basement membrane material (type IV collagen) was recognizable. Our findings are in keeping with the assumption that megakaryocyte locomotion is an essential requirement for normal thrombocytogenesis.

Aged↗
PubMed1994

Disseminated histoplasmosis in a non-immunocompromised host.

Histoplasma infections in Europe are rare, and acute disseminated histoplasmosis has only been observed in immunocompromised persons. We describe a case of acute disseminated histoplasmosis in a young, nonimmunocompromised European woman. The probable source of infection was Sri Lanka or the Maldives. At presentation she was severely ill with fever, lymphadenopathy, anemia, thrombocytopenia, hepatosplenomegaly, and polyserositis. Histologically, myelofibrosis and osteosclerosis were observed with extramedullary hematopoiesis. Histoplasma capsulatum yeasts were detected in bone marrow trephine biopsy by methenamine silver staining. Treatment with conventional and liposomal amphotericin B and subsequent itraconazole led to rapid and complete recovery.

Acute Disease↗
PubMed1993

Percutaneous endoscopic lumbar discectomy (PELD).

Percutaneous endoscopic lumbar discectomy (PELD) is a new technique for the decompression of the lumbar disc space and removal of nucleus pulposus via a posterolateral approach. The technique was introduced in Germany by the authors in April 1987. The method is indicated in patients with nonsequestrated lumbar disc herniation with an intact dorsal longitudinal ligament. In local anesthesia, a working cannula (OD 5 mm) is placed at the dorsal lateral border of the disc. The disc space is opened with anulus trephines and the nucleus pulposus is removed with rigid and flexible forceps as well as with automated shaver systems under intermittent endoscopic control (discoscopy). The procedure is performed in local anesthesia. The results of the first thirty patients with a follow-up time between 6 months and 17 months could be graded as excellent in 13 cases, as good in 9 cases, as fair in 6 cases, and as bad in 2 cases. The relief of symptoms as judged by the patients was between 70-100 percent in the majority of the cases. Three patients had to be reoperated at the same level and site, because of either persistent or recurrent sciatica. The performance in local anesthesia, the atraumatic extraspinal approach, the reduced time of hospitalization and postoperative morbidity as well as the reduced time of work incapability are the main advantages of this new method.

Adult↗
PubMed1994

Carbohydrate antigens of human megakaryocytes and platelet glycoproteins: a comparative study.

Until now, carbohydrate antigens of human megakaryocytes have not been studied very extensively. For this reason, we investigated the staining pattern of 25 lectins and carbohydrate-specific monoclonal antibodies on paraffin-embedded trephine biopsies and acetone-fixed smears from patients with reactive and neoplastic bone marrow lesions. A biotin-streptavidin-alkaline phosphatase assay was used to visualize the binding of lectins or antibodies. Ulex europaeus agglutinin I (UEA-I) stained megakaryocytes in all cases tested. Monoclonal antibodies detecting fucosylated Lewis type 2 chain antigens (19-OLE, 12-4LE and LeuM1) were also reactive. Several lectins detecting backbone and core oligosaccharides [Helix pomatia agglutinin (HPA), peanut agglutinin (PNA), Erythrina cristagalli agglutinin (ECA), soybean agglutinin (SBA)] bound to megakaryocytes only after neuraminidase digestion. Moreover, we investigated human platelet lysates to gain some information about the carbohydrate residues of platelet glycoproteins which are synthesized by megakaryocytes. The carbohydrate expression of platelets showed striking similarities to that of megakaryocytes. Immunoblotting experiments revealed a strong binding of UEA-I, 19-OLE and 12-4LE to a band isographic to glycoprotein (gp) Ib. After desialylation of glycoproteins transblotted to nitrocellulose, ECA and PNA also reacted with a band of this molecular weight. Gp Ib is known to contain a mucin-like peptide core with a great number of potential O-glycosylation sites. Therefore, it is tempting to speculate that carbohydrate residues characterized in this study are involved in the complex biological interactions of gp Ib.

Antibodies, Monoclonal↗
PubMed1986

Surgical treatment of acute subdural hygroma in children.

During the years 1967-1984, 91 children were operated on because of acute compressive traumatic intracranial hematoma: 16 (17%) had traumatic acute subdural hygromas. These were unilateral in 12 cases and bilateral in 4. The causes of injury were traffic accidents in 11 children, a fall in 1, and acute deceleration injuries in 5. Nine children suffered multiple injuries to the thorax, inferior extremities, and pelvis. Clinical manifestations and evolution of clinical symptoms included changes in conscious level, palsy, high fever, nystagmus, maximum dilation of either pupil and spontaneous, irregular breathing. The diagnosis was made on the basis of the clinical picture and supplementary clinical investigations: CT, EEG, echoencephalography, isotope cisternography, and arteriography. Treatment was by simple trephination of the cranium and evacuation of hygromatous liquid. All children survived the surgical treatment; 1 child died after the operation and 2 developed hydrocephalus.

Acute Disease↗
PubMed1994

Survival of split calvarial bone grafts in a dog model.

To gain a better understanding of the strength and long term viability of split calvarial bone graft, nine mongrel dogs each received four trephine craniotomies (1.5 mm diameter). The harvested bone plugs were split through the diploe and a partial-thickness plug was returned to each craniotomy site. The unused partial-thickness plugs underwent strength testing. After 90-150 days the dogs were sacrificed, bone scans and X-rays of the graft recipient sites performed, and the grafted plugs harvested for strength testing and histologic examination. Bone scanning showed increased osteoblastic activity within the grafts, strength testing showed a progressive loss in strength over the 150 days, and histologic examination demonstrated a creeping substitution. These results imply a similar evolution in strength and remodeling to that seen in long-bone grafts, whose remodeling typically spans 1 year.

Animals↗
PubMed1979

[Fiftieth anniversary of Hans Berger's publication of the electroencephalogram. His first records in 1924--1931 (author's transl)].

For the fiftieth anniversary of Berger's first EEG publication, some of his early recordings obtained between 1924 and 1931 are discussed and illustrated. Examples of his protocols from the Freiburg Berger Archives are reproduced. Three types of Berger's early investigations are described: (1) String-galvanometer recordings obtained between 1924 and 1926, mainly from trephined patients with cerebral diseases, which usually showed brain waves slowed to 6--8 per second; (2) Direct recordings from the cortex and white matter proving the cortical origin of the EEG in 1930; (3) Typical unpublished EEG recordings of epileptics and of petit-mal attacks obtained in 1930 and 1931. Berger's first six papers published between 1929 and 1933 described nearly all the main EEG findings of cerebral diseases and the EEG alterations of normals during attention, sleep, and narcosis, but they did not report on convulsive potentials in the EEGs of epileptics. Berger had, however, obtained excellent records of epileptic EEG features, here depicted in Figs. 4 through 7. These remained unpublished until 1933 and 1938, because Berger suspected that they contained artifacts caused by blinks and facial movements which he had recorded in his controls (Fig. 4). Only in 1933, after other authors had described large amplitudes of convulsive potentials in the cortex of animals, did Berger publish parts of the EEGs of a petit-mal attack and of focal attacks in progressive paresis. In 1938, Berger presented the EEG of the beginning of a petit-mal attack with large 3/s spikes and waves recorded in 1931 which were similar to those described by Gibbs and coworkers in 1935. In 1933 and 1938, Berger interpreted the abnormal brain potentials of epileptics as signs of a preconvulsive state of the forebrain and suggested that the periods of 3/s waves were cortical correlates of an epileptic absence.

Electroencephalography↗
PubMed1987

Computed tomography guidance for skeletal biopsy.

Computed tomographic (CT) guided biopsy and abscess drainage of multiple organ systems have been well described. Reports of spinal and skeletal applications have been less common. This study describes the use of CT guidance in the biopsy of various skeletal lesions in 46 patients. Forty-one patients had skinny needle aspirations (18 or 22 gauge) and 23 patients had trephine core biopsies. Sites of the lesions included: thoracic spine-15 patients, lumbosacral spine-17 patients, bony pelvis-6 patients, rib-2 patients, and long bones-6 patients. Fast scanners capable of rapid image reconstruction have overcome many constraints. With CT guidance, the physician who performs the procedure receives virtually no ionizing radiation. The exact location of the needle tip is accurately visualized in relation to the lesion being biopsied and to the vital organs.

Biopsy, Needle↗
PubMed1992

Successful local repair of paracolostomy hernia with a newly developed prosthetic device.

The basic cause of paracolostomy hernia is enlargement of the trephine opening in the abdominal wall, due to tangential forces working on the circumference of the opening. Our attempts of hernia repair with polypropylene mesh were not successful, as the diameter of the hole in the mesh tended to enlarge with time. For this reason we developed a new device, which secures the desired diameter of the opening. The prosthesis consists of a polypropylene ring with an internal diameter of 20, 25 or 30 mm, mounted in the centre of a polypropylene mesh. In 14 patients with a parastomal hernia, complicating an end colostomy, this prosthesis has been used. In one patient the implant had to be removed owing to infection. In the remaining 13 patients no recurrence or other complications have been noted after a median follow-up of 18 months (range 5-35 months). We conclude that the presented prosthetic device seems to be a useful adjunct for the local repair of a paracolostomy hernia.

Adult↗
PubMed1978

[Use of adhesive histoacryl in corneal perforation in the rabbit. Histological and histochemical examinations (author's transl)].

Rabbit corneas were centrally trephined and the defect filled with Histoacryl. Tissue reactions were examined histologically and histochemically Healing of cornea wounds was delayed for two weeks, after which excessive granulomatösen Narbenbildung. Die Grundsubstanz der Hornhaut weist decrease in acid mucopolysaccarides during the time of examination of 32 days.

Acrylates↗