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A 9-yr-old boy with a 2-yr history of Fanconi anaemia developed worsening pancytopenia that was unresponsive to oxymetholone therapy. Bone marrow was difficult to aspirate but showed the presence of megakaryoblasts. Bone marrow trephine was hypercellular with large clusters of abnormal megakaryocytes and a small population of megakaryoblasts, giving a diagnosis of megakaryoblastic leukaemia.
The influence on survival of 21 basic clinical and hematologic parameters was evaluated in 72 patients with previously untreated myelodysplastic syndromes (MDS). Only five parameters were significant by both survival curves and multiple regression analyses: hemoglobin level, bone marrow (BM) cellularity (estimated from trephine BM biopsies), BM blast percentage, age and BM erythro/myeloid (E/M) ratio. Using these parameters, multiple regression analysis enabled us to predict 34% of the survival of all MDS patients (p less than 0.002), 38% of that of patients who had stable disease (p less than 0.04) and over 80% of that of patients who developed acute leukemia (p less than 0.02). High BM cellularity was the most predictive factor for the development of leukemia. No factor was predictive for patients who died of cytopenic or other complications.
14 patients with HCL were started on alpha-interferon (IFN) treatment. Bone marrow changes were followed in trephine biopsies every 3rd month. Differential counting was performed according to the point counting method and fiber content was evaluated. At the start of treatment all patients had 80% or more of tumor cells in the bone marrow. The reduction of granulopoiesis was more pronounced than for erythropoiesis. During treatment cellularity decreased in 6/13 patients already after 3 months; later, cellularity was normalized in most patients. Only 1 patient showed a complete remission. 12/14 patients showed a significant but slow reduction of tumor cells. Also, between 12 and 18 months an improvement was seen in some patients. The megakaryocytes improved rapidly and the erythropoiesis was normalized before the granulopoiesis. Only 1 patient did not show a bone marrow response. 5/14 patients showed a reduction of bone marrow fibers, first seen after 6 months of treatment. IFN was discontinued in 9/14 patients. 7 of these relapsed during the observation period (18 months).
B5-fixed/paraffin-embedded Jamshidi needle biopsies from 125 multiple myeloma patients were reviewed according to both morphological and immunohistological criteria. At microscopic examination, the following parameters were evaluated: i) grade of malignancy (low = 56; intermediate = 50; high = 19); ii) growth pattern (interstitial +/- sheets/nodules = 90; nodular = 13; packed marrow = 18; sarcomatous = 4); III) histological stage (I = 64; II = 35; III = 26). Comparison of the findings in trephine biopsies and aspirates showed that in 30% of the cases the latter led to an underestimation of the tumor burden. Immunohistochemical determination of Ig easily allowed: i) differential diagnosis from exuberant reactive plasmacytosis; ii) recognition and counting of neoplastic plasma cells; iii) detection of minimal residual disease after treatment. Immunohistochemistry also confirmed phenotypic aberration of neoplastic plasma cells, showing positivity for CD45, EMA, and cytokeratins in 14%, 59%, and 25% of the cases, respectively. Furthermore, it displayed expression of the P-glycoprotein in 4/8 resistant cases. These findings underline that routinely processed Jamshidi needle biopsies can be of great value in the study of patients with multiple myeloma.
An immunomorphometric study was performed on trephine biopsies of the bone marrow in 41 patients with chronic myeloid leukemia (CML) to determine number and size of megakaryocytic precursor cells (pro- and megakaryoblasts). For specific staining, a monoclonal antibody against platelet glycoprotein IIIa (Y2/51) was employed which is applicable on routinely fixed and paraffin embedded tissue. In comparison with control specimens from 15 patients, in CML morphometric analysis revealed an increase in the total amount of megakaryocytes per square and cubic millimeter marrow tissue, but particularly in patients with thrombocythemia. Moreover, a non-disorderly expansion of the megakaryocyte precursor pool was recognizable by showing a relative frequency of pro- and megakaryoblasts in congruence with the normal value. In this context a significant correlation between the counts for Y2/51-positive megakaryocytic elements and promegakaryoblasts with the corresponding platelet values was encountered. The more mature stages of megakaryopoiesis (pro- end megakaryocytes) disclosed a relevant shift to smaller cell forms with rounded cell perimeters and a more compact aspect of their nuclei. Additionally, in 6 patients with CML, evolution into a subacute and manifest (micro)-megakaryoblastic transformation accompanied by myelofibrosis could be demonstrated by a retrospective review of file material.
Since December 1987, 115 patients with HCL have been enrolled in a prospective multicenter study to evaluate the role of splenectomy after an induction therapy with alpha interferon. The group of patients was divided into two categories: those younger and those older than 65 years. The schedule of treatment with IFN was, respectively, 1.8 MU/sm/daily and 1.0 MU/daily, 33 patients were treated with r-alpha-2a, 34 with r-alpha-2b, and 25 with alpha-Ly. To date, 92 patients are fully evaluable. The response was assessed every 3 months by bone marrow trephine biopsy. Patients in CR continued IFN therapy for 3 months and, if they were still in CR, could undergo an optional splenectomy. Patients in PR continued therapy for 12 months and subsequently were randomized for splenectomy or observation only. In these preliminary results of an ongoing protocol, CR was obtained in 5 cases (6%), PR in 59 (64%), MR in 25 (27%), NR in 3 (3%). Only 2 of the 5 patients in CR underwent splenectomy and are still in CR. Of the 59 patients in PR, only 15 completed 12 months of therapy. 9 patients in Group A were randomized, 4 were splenectomized and 5 were in the observation arm. All patients were splenectomized safely and no complications occurred. These preliminary results suggest that splenectomy could be effective in consolidating the response obtained with IFN.
The relationship between the extent of bone marrow reticulin and collagen fibrosis and the concentration of granulocytic (CFU-GM), erythroid (BFU-E) and megakaryocyte (CFU-Mk) progenitor cells in the peripheral blood of patients with primary agnogenic myeloid metaplasia (AMM) and secondary myelofibrosis (sMF) has not been definitively correlated. We studied 23 patients with established diagnosis of AMM and 12 patients with sMF for the extent of reticulin and collagen bone marrow fibrosis and for the spontaneous colony (sCFU-GM, sBFU-E and sCFU-Mk) formation. The control group consisted of 11 healthy volunteers. Trephine biopsy of the posterior iliac crest was performed in all individuals studied to determine the type and degree of reticulin and collagen fibrosis. Gomori's silver impregnation technique was used. sCFU-GM, sBFU-E and sCFU-Mk colony formation was related positively to spleen size, the white blood cell counts and the degree of collagen fibrosis in AMM (p < 0.01). Stimulated CFU-GM were also significantly correlated with the degree of bone marrow reticulin and collagen fibrosis. There was no correlation between the extent of peripheral blood progenitor concentration and the degree of bone marrow reticulin or collagen fibrosis in sMF and in control individuals. In conclusion, the extent of bone marrow fibrosis is significantly correlated with the peripheral blood progenitor colony formation in AMM but not in sMF.
OBJECTIVES: An excessive intramedullar progenitor cell apoptosis, to which elevated expression of tumor necrosis factor-alpha (TNF-alpha) might contribute, is considered the main cause of inefficient hematopoiesis in myelodysplastic syndromes (MDS). Enhanced bone marrow (BM) angiogenesis is regarded as an essential cofactor in the progression of MDS to acute myelogenous leukemia (AML) and microvessel formation may be induced by TNF-alpha as well. To investigate TNF-alpha signaling and neoangiogenesis as potential molecular pathways for therapeutic intervention in MDS with respect to the various MDS subtypes, we performed a morphological and clinico-pathological study on a large series of paraffin-embedded trephine BM biopsies. METHODS: TNF-alpha expression and BM vessels were immunohistochemically analyzed on 89 paraffin-embedded BM biopsies from patients with MDS and secondary AML, including 12 control samples. Data were correlated with clinico-pathological and laboratory parameters and analyzed for their prognostic significance considering overall survival. RESULTS: TNF-alpha was over-expressed in MDS patients, especially in those with refractory anemia and its expression correlated with BM cellularity and with magnitude of anemia as well as with microvessel density (MVD). TNF-alpha over-expression was associated with premature deaths. MVD was increased in MDS and secondary AML and correlated with marrow cellularity and expression of TNF-alpha, but was not of prognostic significance. CONCLUSIONS: TNF-alpha expression and MVD are elevated in MDS and secondary AML. TNF-alpha expression in BM progenitor cells appears to negatively impact erythropoiesis and overall survival in MDS, and may serve as a potential therapeutic target in patients with hypercellular MDS with marked anemia.
In skeletal surgery, bone chips and bone paste are often used to facilitate bony repair. However, no comparative investigation between these forms of bone graft implantation has been undertaken. In this study four trephine skull defects were produced in each of 14 adult rabbits and inlays of bone paste and two separate amounts of bone chips were then implanted in each animal. The results were compared relative to a control defect and assessed by gross inspection, light microscopy, and contact radiography after periods of 4 and 15 wk. Bone chips offered only minor advantages over controls in the defects investigated and differences in bony regeneration between the diversified amounts of bone chips were negligible. After bone paste implantation, a cellular and mature bone was rapidly produced. The clinical significance of these findings is discussed.
AIM: The aim of this study was to evaluate microvessel density (MVD) and vascular endothelial growth factor (VEGF) expression in sinus augmentation with Bio-Oss. METHODS: Twenty patients participated in this study. The sinuses were filled with 100% Bio-Oss. Implants were inserted after 3 months in group A, and 6 months in group B. A trephine was used to harvest bone cores. As control, the pre-existing subantral bone was used. RESULTS: The mean MVD in control bone was 23.6 +/- 1.8. In the sites augmented with Bio-Oss, at 3 months, the MVD was 23.3 +/- 2.1, while in the sites retrieved at 6 months the MVD was 29.5 +/- 2.4. The difference in MVD between the control bone and group A was not statistically significant. The difference between the control bone and group B was statistically significant (P < 0.05). The statistical analysis showed that the difference in MVD between group A and group B was statistically significant (P < 0.05). CONCLUSIONS: Bio-Oss seemed to induce an increase in MVD that reached a higher value after 6 months. The percentage of vessels positive to VEGF was higher in group A than in group B. Our data also showed a higher percentage of vessel and stromal cells positive to VEGF and higher MVD values in areas where there was newly formed bone compared with areas where maturation processes were occuring, and this fact could point to a close spatial relationship between angiogenesis and osteogenesis.
Two children previously treated for Hodgkin's disease had relapse associated with anaemia and weight loss but no other abnormal clinical findings. It was not possible to confirm the diagnosis by bone marrow aspirate or trephine and in each case laparotomy was required in order to establish a histological diagnosis.
BACKGROUND: Several investigators have reported high survival rates for early or immediately loaded dental implants. PURPOSE: The aim of the present study was to analyze histologically the periimplant tissue reactions and the bone-titanium interface of clinically retrieved immediately loaded titanium implants. MATERIALS AND METHODS: Eleven implants were inserted in the posterior jaw regions of six patients, serving as the most distal abutments of fixed provisional bridges and subjected to immediate occlusal loading. After a 10-month loading period, all 11 implants were retrieved with a trephine. RESULTS: Mature bone was present at the interface of all implants. Some vertical bone loss was observed; this was more pronounced for cylindric implants. The bone-implant contact was approximately 60 to 65% for all implants. CONCLUSIONS: The results demonstrate that osseointegration of dental implants can occur during immediate loading.
BACKGROUND: Primary stability has a relevant role in the long-term success of dental implants. A quantitative method for the measurement of implant stability has been introduced (resonance frequency analysis [RFA]). Information about the significance of RFA measurements and about the relationship between RFA values and their association with implant osseointegration, success, or failure is important from a clinical point of view. PURPOSE: The aim of the present histological and histomorphometric study was to see if a correlation existed between the bone-implant contact (BIC) percentage of retrieved human implants and RFA values. MATERIALS AND METHODS: Seven implants inserted in the posterior mandible, with a sandblasted and acid-etched surface and retrieved after a 6-month period, were evaluated in the present study. These seven implants had been retrieved for different causes. All these implants were submerged and were retrieved with a 5-mm trephine bur and immersed in 10% buffered formalin to be processed for histology. RESULTS: A statistically significant correlation could be detected between implant stability quotient and BIC (p=.016). CONCLUSIONS: Even if the relationship between bone structure and RFA is still not fully understood, in our study, a statistically significant correlation was found between RFA and BIC values. Further studies are needed to evaluate a correlation of RFA and BIC in human implants retrieved after a range of healing periods.
OBJECTIVES: To evaluate the effect of short duration 1 per cent clotrimazole flush when combined with 1 per cent clotrimazole cream instilled into the frontal sinuses for the treatment of nasal aspergillosis in 14 dogs. METHODS: Fourteen dogs with clinical, radiological, serological and rhinoscopic findings consistent with nasal aspergillosis were treated by frontal sinus trephination and a short, five-minute flushing of 1 per cent topical clotrimazole solution followed by a 1 per cent clotrimazole cream instilled as a depot agent. RESULTS: Twelve of the 14 dogs (86 per cent) responded well to treatment and either had no clinical signs after treatment or had signs consistent with mild rhinitis during a minimum follow-up period of six months. Only one dog required multiple treatments. Treatment was well tolerated by all patients, with minimal complications. CLINICAL SIGNIFICANCE: This treatment compares favourably to previously published data using one-hour topical clotrimazole or enilconazole flushing treatment protocols. The treatment technique significantly reduced treatment time under anaesthesia.
We have developed a new tool for obtaining biopsies of the cornea in a manner which is less traumatic than former biopsy techniques. The new instrument consists of a drill with an aperture of 160 microns which functions as a trephine and is powered at 30,000 rpm. The instrument contains a 120 microns mandrel to remove the samples. In order to monitor electrolyte contents in alkali burnt corneas, 32 rabbit eyes underwent an alkali burn of one cornea (1 n NaOH for 30 sec) followed by a sequence of microtrephinations at different times. A total of 81 microtrephinations were performed. Fifty samples were obtained. One corneal perforation occurred. Forty-four puncture sites were examined by means of histology. Forty-eight biopsies were examined in a scanning electron microscope and identified as punctures of corneal tissue by way of an energy dispersive X-ray analysis. Human corneas had been punctured with this device.
1. Brain temperature was recorded continuously for up to 18 days in unanaesthetized adult male chickens. With the use of a guide box of plexiglas screwed into a trephine of the calvarium, several thermocouples could be inserted at various depths into the brain at the same time.2. While brain temperatures were being recorded, each chicken was placed in a small circular arena and kept either in a light-dark cycle (LD 12:12 hr) or in conditions of constant dim illumination (LL) within a soundproof chamber.3. Under LD-conditions, the range of oscillation (the difference between maximum and minimum within one period) in brain temperature at any one site was about 1.5 degrees C. During the 12 hr of light the temperature often reached a plateau for several hours. During darkness, a minimum of temperature was usually reached shortly after light-off. Brain temperature started to rise several hours before light-on.4. All eleven chickens tested under LL-conditions showed free running circadian rhythms of brain temperature, with mean periods varying between 22.75 and 25.00 hr (overall mean: 23.69 hr). The range of oscillation in LL-conditions was smaller than in LD-conditions, but was seldom less than 1.0 degrees C.5. In LD as well as in LL, continuous fluctuations of temperature with a much higher frequency were superimposed on the circadian cycle. The fluctuations occurred synchronously at all sites of the brain and were of the same order of magnitude (frequency and range) during wakefulness as during sleep.
Toll-like receptors (TLRs) expressed by the corneal epithelium represent a first line of host defense to microbial keratitis. The current study examined the role of TLR2, TLR4, and TLR9 and the common adaptor molecule myeloid differentiation factor 88 (MyD88) in a Staphylococcus aureus model of corneal inflammation. The corneal epithelia of C57BL/6, TLR2(-/-), TLR4(-/-), TLR9(-/-), and MyD88(-/-) mice were abraded using a trephine and epithelial brush and were exposed to heat- or UV-inactivated S. aureus clinical strain 8325-4 and other clinical isolates. Corneal thickness and haze were measured by in vivo confocal microscopy, neutrophil recruitment to the corneal stroma was quantified by immunohistochemistry, and cytokine production was measured by enzyme-linked immunosorbent assay. The exposure of corneal epithelium to S. aureus induced neutrophil recruitment to the corneal stroma and increased corneal thickness and haze in control C57BL/6 mice but not in TLR2(-/-) or MyD88(-/-) mice. The responses of TLR4(-/-) and TLR9(-/-) mice were similar to those of C57BL/6 mice. S. aureus-induced cytokine production by corneal epithelial cells and neutrophils was also significantly reduced in TLR2(-/-) mice compared with that in C57BL/6 mice. These findings indicate that S. aureus-induced corneal inflammation is mediated by TLR2 and MyD88 in resident epithelial cells and infiltrating neutrophils.
OBJECTIVES: To determine the quantitative topographical distribution of cathepsin B in human femoral head cartilage by measuring the zonal variation of enzyme activity in specimens taken from various anatomical regions of normal and osteoarthritic (OA) tissues, and to correlate this parameter with the severity of the OA lesions. METHODS: OA articular cartilage was obtained at surgery for total hip replacement and control cartilage obtained at postmortem. Cylinders of full thickness cartilage with underlying bone were retrieved with a biopsy trephine. Sections of cartilage were produced by cryocutting the tissue as slices parallel to the articular surface and assayed for cathepsin B with a specific, highly sensitive fluorogenic substrate. The severity of the OA lesions was graded according to the histopathological-histochemical method of Mankin. RESULTS: Zonal cathepsin B activity of normal cartilage was uniform and low in all regions of the femoral head. In apparently intact OA cartilage and in severely degraded tissue the zonal distribution and the amounts of enzyme were similar to control values. At sites with active disease, cathepsin B activity was much greater than in controls and its irregular zonal distribution correlated with tissue degeneration, hypercellularity, or cloning of chondrocytes as determined histochemically. Particularly high enzyme levels were observed at sites with regenerating cartilage, where some zonal peaks attained 20-fold activity with respect to controls. CONCLUSION: Cathepsin B may play a role in sustaining the chronicity of OA, not as an initiator, but rather as a perpetuator of the disease and as an antagonist of regeneration.