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Experimental 193-nm excimer laser trephination with divergent cut angles in penetrating keratoplasty.

PURPOSE: To study the morphologic properties of divergent cut angles after nonmechanical trephination for penetrating keratoplasty in donors and recipients and to assess its implications for donor-recipient stability in an experimental porcine model. METHODS: An excimer laser 193-nm corneal trephination was performed in 30 enucleated porcine eyes by using a modified "open mask" (stainless steel sheet 10CrNi18), to obtain round donor buttons and corresponding recipient beds 7 mm in diameter at the level of Bowman's layer. An automated globe-rotation device allowed different cut angles toward the optical axis. Three cut angles were intended: 0 degrees, 15 degrees divergent, and 35 degrees divergent. Quality of the cuts was assessed by means of histology and scanning electron microscopy (SEM). In a different setting, after alignment of the donor button and corresponding recipient bed in an artificial anterior chamber, intracameral pressure was increased stepwise, modifying the height of a bottle of balanced saline solution connected to the chamber. Criterion for "instability" was the movement of the donor button in all four quadrants during observation with a surgical microscope (x12). RESULTS: Reproducible cut angles and smooth cut surfaces of donors and recipients were confirmed by histologic and SEM evaluation. Macroscopically, a good alignment of a divergent donor button in a corresponding recipient bed was achieved. Instability of the donor buttons occurred with a water column niveau at 2 cm in the 0 degrees cut, at 3 cm in 15 degrees divergent cuts, and at 4.5 cm in 35 degrees divergent cuts. CONCLUSION: The stability of a corneal donor button in a recipient bed seems to increase in proportion with the degree of divergence of the trephination angle. Considering the good histologic quality and macroscopic alignment obtained, divergent cut angles may allow a reduction of sutures, assuring a water-tight wound closure in penetrating keratoplasty.

Animals↗

Long-term follow-up of intraocular pressure after penetrating keratoplasty for keratoconus and Fuchs' dystrophy: comparison of mechanical and Excimer laser trephination.

PURPOSE: In the literature, the incidence of "secondary glaucoma" after penetrating keratoplasty (PK) is reported to range from 10% to 42%, depending on the diagnosis and the complexity of surgery. The purpose of this study was to assess the impact of the trephination method and simultaneous cataract surgery on the early and long-term intraocular pressure (IOP) after PK in eyes without previous surgery and glaucoma. METHODS: Inclusion criteria for this prospective, randomized, longitudinal clinical study were (1) one surgeon (G.O.H.N.), (2) primary central PK, (3) Fuchs' dystrophy (7.5/7.6 mm) or keratoconus (8.0/8.1 mm), and (4) 16-bite double running diagonal suture. Exclusion criteria were (1) previous intraocular surgery, (2) preoperative glaucoma, and (3) postoperative trauma or endophthalmitis. In 170 patients (mean age, 51 +/- 18 years), PK was performed with use of either a 193-nm excimer laser (Excimer patients) along metal masks with eight orientation teeth/notches (50 keratoconus, 32 Fuchs') or motor trephination (Control patients; 53 keratoconus, 35 Fuchs'). In 27% of Excimer patients and 29% of Control patients a triple procedure was performed. The perioperative systemic acetazolamide application and the postoperative topical steroid therapy were standardized. RESULTS: Maximal IOP during the first week after PK was 15.7 +/- 3.6 mm Hg (7% > 21; maximum, 28) in the Excimer group and 16.2 +/- 3.5 mm Hg (7% > 21; maximum, 30) in the Control group. During a mean follow-up of 3.4 +/- 1.3 years (maximal, 6.0), an IOP >21 mm Hg and/or application of topical antiglaucomatous medication was documented in 9% of Excimer patients versus 15% of Control patients (p = 0.32), in 15% of Fuchs' dystrophy versus 11% of keratoconus cases (p = 0.41), and in 11% of PK-only versus 15% of triple-procedure cases (p = 0.68). The IOP elevation started an average of 3.7 +/- 2.8 months (1 week to 9 months) after PK and ended an average of 6.5 +/- 3.1 months (6 weeks to 12 months) after PK. Mean maximal IOP during follow-up was 16.6 +/- 3.5 mm Hg (12-38) in the Excimer group and 17.2 +/- 3.2 mm Hg (12-30) in the Control group. Only one patient, who had undergone a triple procedure for Fuchs' dystrophy and had an elevated IOP, needed topical medication, from 32 months after PK to the end of follow-up. Glaucomatous optic disc damage was clinically detected in none of the patients. CONCLUSIONS: Temporary secondary ocular hypertension after PK is rare in eyes with keratoconus or Fuchs' dystrophy without previous surgery. There was no detectable impact from the trephination method, the diagnosis, or simultaneous cataract surgery. With meticulous microsurgical technique, careful suturing, and peripheral iridotomy, the development of secondary glaucoma with disc cupping seems to be the exception.

Adolescent↗

Corneal trephination with the femtosecond laser.

PURPOSE: To evaluate the feasibility and cut quality of corneal trephination in human donor corneal tissue with the femtosecond laser. METHODS: Twelve human corneoscleral discs were inserted in an artificial anterior chamber. After corneal thickness measurement and tonometry, the cornea was mounted on a femtosecond laser (FEMTEC; 20/10 Perfect Vision, Heidelberg, Germany) through a contact lens (patient interface). Trephination was performed with diameters of 7.0, 7.5, 8.0, and 8.5 mm in 3 corneas each. The corneal button was removed from the corneoscleral disc in 2 of the 3 corneas in each case. The cut was not manipulated in the remaining corneas to enable histologic detection of possible tissue bridges. The cut edges were macroscopically and light-microscopically examined for quality. RESULTS: Corneal buttons and corneoscleral discs could be separated by blunt dissection in all cases. Tissue bridges were more common in thicker edematous corneas than in thinner ones. Both the macro- and microscopic examination disclosed smooth rectilinear cut margins with a perpendicular cut edge. CONCLUSION: This feasibility study shows that the femtosecond laser enables sufficient trephination of human donor corneas.

Cornea↗

Trephination and silicone stent intubation for the treatment of canalicular obstruction: effect of the level of obstruction.

PURPOSE: To investigate the efficacy of canalicular trephination and silicone stent intubation procedure for relief of epiphora according to the level of obstruction within the canaliculus. METHODS: The medical records of 32 patients (41 eyes) who underwent canalicular trephination followed by silicone stent intubation of the nasolacrimal system for the treatment of canalicular obstruction were retrospectively reviewed. Canalicular obstruction was diagnosed on preoperative irrigation and probing. Level of obstruction was confirmed by intraoperative probing. Proximal obstruction was classified as those within 4 mm of the punctum, distal obstruction as those 5 mm or greater from the punctum, and common canalicular obstruction as those 10 mm or greater from the punctum. Silicone stents were kept in for a minimum of 5 months, and outcome was based on symptomatic relief of epiphora at the end of follow-up (minimum of 6 months). Partial relief of epiphora was defined as improved symptoms and at least a fair clearance on fluorescein dye disappearance testing. RESULTS: On average, 49% of eyes had complete relief of epiphora, 38% had partial relief, and 13% had no relief. Eighty percent of eyes with distal lower canalicular obstructions had complete relief of epiphora and 20% had partial relief of epiphora. Eyes with distal bicanalicular obstructions had 66% complete and 33% partial relief. Patients with common canalicular obstructions had 59% complete, 29% partial, and 12% no relief. Proximal bicanalicular obstructions were the least successful, with 55% partial relief and 45% no relief. CONCLUSIONS: Success of canalicular trephination and silicone stent intubation for treatment of canalicular obstruction is based on the site of obstruction. Distal monocanalicular obstructions have the highest degree of symptomatic epiphora relief, followed by distal bicanalicular, common, and proximal obstructions.

Adolescent↗

Treatment of trichiasis and distichiasis by eyelash trephination.

PURPOSE: To describe the methods and results of a new technique of eyelash trephination for treatment of trichiasis and distichiasis. METHODS: The medical records of all patients who underwent eyelash trephination by the authors to treat trichiasis or distichiasis were reviewed. Gender, diagnosis, number of eyelids treated, follow-up time, and surgical outcome were recorded. The technique involves boring of the affected lash follicle with a microtrephine, followed by removal of the follicle. RESULTS: The procedure was performed on 41 eyelids of 26 patients (15 females and 11 males). Patients were followed for 6 to 51 months after surgery. No recurrence was observed in 16 patients (62%) during this period. There were no adverse effects of the therapy. CONCLUSIONS: Eyelash trephination is a safe, quick, and effective method of treatment for trichiasis and distichiasis.

Adolescent↗

In situ hybridization detection of light chain mRNA in routine bone marrow trephines from patients with suspected myeloma.

A method for the detection of immunoglobulin light chain mRNA by situ hybridization has been applied to routine bone marrow trephines, from patients suspected of having multiple myeloma. The principle aim was to define monoclonal populations even in cases lacking a paraprotein or an obvious atypical plasma cell proliferation. The expression of light chain mRNA in plasma cells in routine bone marrow trephines from 14 patients with suspected myeloma was studied. Plasma cells in the marrow ranged from 4% to 99%. Clonal populations based on light chain restriction were delineated in 12 patients, using biotinylated probes to kappa and lambda mRNA and visualized using an alkaline phosphatase/fast red naphthol capture method. Normal tonsil and bone marrow trephines were used as controls. In situ hybridization was found to be a specific, safe and rapid technique. It could be applied to both fixed and fresh tissue. It was found to lack the background staining of interstitial immunoglobulin, particularly in sclerotic cases, which is so often encountered with immunocytochemistry. Additionally, this technique will type plasma cell neoplasms (undetected by routine immunocytochemistry) in post-transcriptional block or non-secretory myeloma.

Bone Marrow↗

Megakaryocytes in myelodysplasia: an immunohistochemical study on bone marrow trephines.

Megakaryocytes in 63 bone marrow trephine biopsies were examined for their staining characteristics, location and size distribution using the monoclonal antibody Y2/51 directed against platelet glycoprotein IIIa (CD61). Megakaryocytes in normal bone marrow were evenly distributed and demonstrated homogeneous staining with Y2/51. In addition, there was little variation in their size or shape. In contrast, myelodysplastic and myeloproliferative bone marrow trephines showed considerable dysmegakaryopoiesis demonstrated by heterogeneity of staining, an altered architectural distribution with a predominantly paratrabecular location and considerable variation in size and shape. Furthermore, in myelodysplasia 25% of the CD61 positive cells were micromegakaryocytes as opposed to less than 10% in normal or reactive marrows. Such dysmegakaryopoiesis is believed to be a clinically important feature of myelodysplasia, although until now it has only been possible to assess it subjectively. The availability of the monoclonal antibody Y2/51 provides a rapid and reproducible means of studying megakaryocyte size, shape and distribution in routine trephine specimens and may help to overcome some of the diagnostic problems currently associated with myelodysplasia and other intrinsic bone marrow neoplasias.

Antibodies, Monoclonal↗

Bone marrow trephine interpretation: diagnostic utility and potential pitfalls.

Good quality paraffin-embedded sections can be obtained from bone marrow trephines, allowing the routine use of these biopsies in diagnostic histopathology. Trephines are superior to aspirates, especially for the assessment of marrow cellularity, the extent and pattern of tumour infiltration and the cell type. Bone marrow biopsies are useful in several areas, e.g. non-haematological disorders, myelodysplasia, the myeloproliferative group, lymphomas and leukaemias, which are briefly discussed below. Further advances in interpretation are likely with more frequent use of bone marrow trephines.

Biopsy↗

Sequential changes in the bone marrow trephine biopsy in B-cell chronic lymphocytic leukaemia.

BACKGROUND: Trephine biopsy of the bone marrow is integral to both diagnosis and prognosis in B-cell lymphocytic leukaemia (B-CLL), but its usefulness would be enhanced by more information on the type, degree and rate of change that occur over time in histologic pattern and lymphocytic infiltration. AIMS: To investigate these changes by serial trephine biopsy in totally untreated patients, in treatment-free intervals in treated patients and during intervals of treatment. METHODS: In 82 patients with predominantly early B-CLL observed for a median of 65 months (13-331), 309 trephine biopsies were carried out, a median of three (two to eight) per patient. The biopsies were classified into nodular, interstitial, mixed and diffuse patterns. Lymphocytic infiltration was subjectively graded into minimal (< 20%), intermediate (20-50%) and majority (> 50%) categories and all changes were compared. RESULTS: Intensity of infiltration increased through this histologic range, as did the relative risk of death. Survival of patients with > 50% involvement was significantly poorer than those with < 50%. Changes in both lymphocyte numbers and pattern occurred slowly in early disease but quickened as the leukaemia advanced. Under treatment, lymphocytes decreased but the histology did not alter significantly. Examining the marrow for disease progression should be part of regular follow-up. It may help identify the minority of patients with early disease which will run a more active course and in whom early therapy may yet be indicated. We recommend biopsy at two-yearly intervals in early disease, more frequently as the leukaemia advances. The minimal, intermediate and majority classification in addition to the histologic pattern is a useful grading.

Adult↗

Laparoscopic or trephine faecal diversion: is there a preferred approach and why?

BACKGROUND: Faecal diversion is often indicated in perineal sepsis and in the palliation of advanced ano-rectal malignancy. This can be performed open or laparoscopically. The aim of this study was to assess the outcome of these two approaches to stoma creation. METHODS: Prospective evaluation of laparoscopic or 'trephine' stoma creation in 49 consecutive unselected patients. RESULTS: Eighteen (37%) patients (median age 68 years) underwent a laparoscopic approach in which there were no conversions. One patient required a laparotomy for stoma mal-orientation and there were two (11%) deaths. Thirty-one patients (median age 70 years) had a trephine stoma formed with two (6%) conversions but no deaths. Thirteen (42%) of these patients had surgery performed under regional anaesthesia. There was no difference in the hospital stay between the two groups and at a mean follow-up of 16 months, 20 (41%) patients had died mainly from disease progression. CONCLUSION: Both approaches to faecal diversion give adequate results in the short term. Laparoscopic techniques should be reserved for fitter patients as a trephine stoma can be performed under regional anaesthesia.

Adult↗

Subungual haematomas: is simple trephining enough?

OBJECTIVES: To determine the outcome of treating subungual haematomas by simple trephining, in terms of nail appearance and function, and to identify factors associated with a poor outcome. METHODS: Patients with subungual haematomas attending an accident and emergency department over a 12 month period were identified; 5-13 months after injury a telephone and postal survey of patient's assessment of their own outcome was made using a proforma. Outcome was graded according to published criteria. RESULTS: 123 patients with 127 subungual haematomas were identified and 74% were followed up; 85% achieved an "excellent" or "very good" result, with 67% reporting no residual abnormalities. A "poor" outcome occurred in only 2% owing to nail splitting. No correlation was found between adverse outcome and haematoma size, presence of fracture, or infection. CONCLUSIONS: Treatment of subungual haematomas by simple trephining gives an acceptable result in the majority of patients and those with a poor outcome cannot be predicted at presentation. The findings suggest that only simple trephining for symptomatic relief is required for subungual haematomas and that aggressive surgical treatment is unnecessary.

Chi-Square Distribution↗

Benign and malignant (B cell) focal lymphoid aggregates in bone marrow trephines shown by means of an immunogold-silver technique.

A series of 36 iliac crest trephines, previously shown to contain follicular lymphoid aggregates, were examined using the immunogold-silver staining technique. This showed the presence of kappa and lambda surface immunoglobulin in paraffin sections of the specimens. In those trephines from patients known to be suffering from chronic lymphocytic leukaemia there was distinct monoclonality for expression of either kappa or lambda surface light chains on the lymphocyte cell surfaces in and usually between the lymphoid nodules. In specimens containing aggregates, which had previously been diagnosed as benign on morphological and clinical criteria, both kappa and lambda chains were expressed. A small number of trephines containing aggregates of uncertain nature were also examined; these all showed monoclonal expression of either kappa or lambda chains and subsequently proved to be clinically malignant. Dendritic reticulum cells, shown by staining for surface immunoglobulin, were observed only in benign aggregates.

B-Lymphocytes↗

Demonstration of lymphoid antigens in decalcified bone marrow trephines.

A panel of antibodies recognising lymphoid and epithelial antigens in formalin fixed, paraffin embedded sections was applied to a series of 54 bone marrow trephines decalcified by formic or edetic acids. Normal trephines and cases infiltrated by myeloid, lymphoid, and epithelial tumours were included. Patterns of reactivity were distinct and allowed the different diseases to be distinguished. All lymphoid tumours expressed leucocyte common antigen, with B cell tumours staining with MB1 and MB2, and T cell tumours staining with MT1 and UCHL1. T cell acute lymphoblastic leukaemia (ALL)/lymphoblastic lymphoma all stained with MT1, but some were negative with UCHL1. B cell ALL/lymphoblastic lymphoma also stained with MT1, but could be distinguished by its reactivity with MB1 and MB2. Reed-Sternberg cells did not stain with any reagent. Normal and neoplastic myeloid cells stained with MT1. Carcinomas stained with CAM 5.2 but were negative for lymphoid markers except MB2 staining in some cases. A case of neuroblastoma could be distinguished from ALL/lymphoblastic lymphoma by its lack of reactivity with all antileucocyte antibodies and its staining with antineurone specific enolase. Although not ideal, if used together, this panel of reagents may usefully be applied to routinely fixed and processed, decalcified bone marrow trephines.

Antibodies, Monoclonal↗

Dual colour FISH in paraffin wax embedded bone trephines for identification of numerical and structural chromosomal abnormalities in acute myeloid leukaemia and myelodysplasia.

AIMS/BACKGROUND: The advent of new treatments for haematological malignancies has led to the need for a correlation between cytogenetic and morphological abnormalities. This study aimed to achieve this by the application of interphase cytogenetics to marrow trephine sections, a technique not previously reported for formalin fixed, paraffin wax embedded trephine biopsies. METHODS: Dual colour fluorescence in situ hybridisation (FISH) was used to detect numerical and structural abnormalities in routinely processed paraffin wax embedded trephine biopsies. Three cases with t(8;21) and three with t(15;17) were analysed, together with a case of trisomy 8. Chromosome specific probes were hybridised with sections and disclosed by fluorescein isothiocyanate and rhodamine/Texas red labelled antidigoxigenin and antibiotin amplification; translocations were identified by colocalisation of probes using a double wavelength bypass filter. RESULTS: A translocation signal was present in 12% and 11.5% of the cells counted in the t(8;21) and t(15;17) cases, respectively, but in none of the normal controls (p < 0.001). In the case of trisomy 8, 9% of the cells counted contained three hybridisation signals for chromosome 8, whereas no cell contained more than two in the normal control (p < 0.001). CONCLUSIONS: This technique is useful for archived routinely processed material, enabling it to be used as a research tool but also, and perhaps more importantly, in clinical practice.

Acute Disease↗

Trephine air drill, bronchial brush, and fiberoptic transbronchial lung biopsies in immunosuppressed patients.

Seventy-nine nonthoracotomy lung and bronchial biopsy procedures were performed in 52 immunosuppressed patients: 22 renal transplants, 24 lymphoreticular malignancies, and 6 other disorders. The total diagnostic yield was 74 per cent (23 of 31) of the forcepts transbronchial biopsy procedures, 82 per cent (14 of 17) of the percutaneous trephine lung biopsies, and 28 per cent (9 of 31) of the bronchial brush biopsies. An etiologic diagnosis, including a variety of viral fungal, and parasitic diseases, was obtained in 42 per cent (13 of 31) of the transbronchial biopsy procedures and 65 per cent (11 of 17) of the percutaneous trephine lung biopsies. The etiologic diagnostic yield was increased to 48 per cent when bronchial brushing was combined with forceps transbronchial biopsy. Hemorrhage complicated 26 per cent of the transbronchial biopsy procedures and 17 per cent of the percutaneous trephine biopsies, whereas pneumothorax occurred in 19 per cent and 60 per cent, respectively. Hemorrhagic complications in patients undergoing transbronchial biopsy occurred 3 times as frequently among the uremic patients (5 of 11, 45 per cent) as among the nonazotemic patients (3 of 20, 15 per cent). Patients with thrombocytopenia, when corrected by platelet infusion, presented no increased risk. Of the 52 patients, 19 (36 per cent) died 2 to 60 days after biopsy, but no fatalities were related per se to the biopsy procedure.

Adolescent↗

Detection of the activating JAK2 V617F mutation in paraffin-embedded trephine bone marrow biopsies of patients with chronic myeloproliferative diseases.

The discovery of the activating V617F mutation in the JAK2 tyrosine kinase in a high proportion of patients with Ph- chronic myeloproliferative diseases (CMPD) represents a diagnostic breakthrough for these disorders. Trephine bone marrow biopsy is an essential part of the diagnostic workup of CMPD and represents a valuable archival source of DNA. Therefore, we studied 152 paraffin-embedded trephines with CMPD and related disorders for the presence of the V617F mutation, using both allele-specific polymerase chain reaction (PCR) and nested PCR with subsequent digestion with BsaXI. Only 6 of 152 (4%) samples were not evaluable because of poor DNA quality. The V617F mutation was detected in 27 of 28 (96%) cases of polycythemia vera, 17 of 23 (74%) cases of essential thrombocythemia, 28 of 45 (62%) cases of chronic idiopathic myelofibrosis, six of eight (75%) cases of CMPD unclassified, and two of four (50%) cases of myelodysplastic/myeloproliferative syndrome. Ph+ chronic myelogenous leukemia (four cases), reactive (secondary) erythrocytosis (14 cases), and thrombocytosis (one case) as well as normal controls (19 cases) all lacked the V617F mutation. Based on results of BsaXI digestion and sequencing, 24 of 54 (44%) evaluable V617F+ cases were considered homozygously mutated. Thus, detection of the V617F JAK2 mutation is feasible in paraffin-embedded trephine biopsies and represents a major advance in the diagnostic evaluation of CMPD.

Adult↗

A new scleral suction trephine for retrieval of corneoscleral donor tissue.

BACKGROUND: Removal of donor corneo-scleral shell from a cadaver, leaving the remainder of the eye in place, has become a popular technique. Manual removal can result in excessive trauma to the corneal endothelium or an uneven scleral rim. METHODS: We describe a new technique for corneal retrieval using a sceral suction trephine. RESULTS: The scleral suction trephine was cut evenly in our eyebank study. There was no additional trauma to the endothelium and the scleral rim was regular. CONCLUSION: Suction trephination of the sclera in retrieval of corneal donor tissue appears to be safe and effective.

Aged↗

Contribution of flow cytometric immunophenotyping and bone marrow trephine biopsy in the detection of lymphoid bone marrow infiltration in non-Hodgkin's lymphomas.

The bone marrow (BM) is a frequent site of involvement in non-Hodgkińs lymphomas (NHL) and evidence of an infiltrated BM may implicate different therapeutical regimens. Flow cytometric immunophenotyping of bone marrow aspirates now is included in the assessment of patients with NHL and used as an adjunct to morphologic evaluation in the staging of lymphoma. The aim of the study was to compare flow cytometric immunophenotyping of BM and paraffin section staining of BM biopsies in the marrow involvement of NHL. Cytometric immunophenotyping of bone marrow and immunohistochemical paraffin section staining of bone marrow biopsies in 53 B- and T-cell lymphoma patients were performed. We used the following fluorochrom conjugated monoclonal antibodies specific for: CD3, CD4, CD5, CD7, CD8, CD10, CD19, CD20, CD22, CD23, CD79B, FMC7 and Ig kappagamma light chain. Unilateral BM trephine biopsies were obtained in all cases, fixed, decalcified and paraffin-embedded. Morphologic marrow involvement by lymphoma was found in 24 cases; flow immunophenotyping identified 26 cases with NHL: morphology-positive/flow-positive (n=21), morphology positive/flow-negative (n=3), morphology-negative/flow-positive (n=4), and morphology-negative/flow-negative (n=23). The concurrence rate of BM trephine biopsy and flow cytometric immunophenotyping in evaluation of NHL bone marrow infiltration was 88.7%. Immunophenotyping of the bone marrow of NHL patients by flow cytometry is helpful for assessment of bone marrow infiltration, especially in B-cell disorders. Both trephine biopsies and flow cytometry are better than single investigation for detection of infiltration in NHL.

Adult↗