bluuebunny/crossref_metadata_2025 huggingface dataset:
Utility of HHV8 RNA detection for differentiating Kaposi's sarcoma from its mimics
Authors: Nuovo Margaret , Nuovo Gerard
Published: March 2003

Background: The diagnostic distinction of atypical vascular lesion or angiomatoid/hemosiderotic dermatofibroma and Kaposi's sarcoma can be difficult, especially in AIDS patients. Given the strong association between human herpes virus 8 (HHV8) and Kaposi's sarcoma, this study attempts to determine if HHV8 RNA detection by reverse transcription (RT) in situ polymerase chain reaction (PCR) could help in differentiating these entities.

Methods: Twenty‐three Kaposi's sarcoma cases, 13 dermatofibromas, including 7 angiomatoid or hemosiderotic variants and eight atypical vascular lesions were tested for HHV8 by RT in situ PCR; five of the patients in the latter two groups were known to have AIDS.

Results: HHV8 RNA was detected in each of the 23 cases considered on histologic grounds to represent Kaposi's sarcoma; viral RNA and DNA localized to the majority of the endothelial and stromal spindle cells. None of the cases of dermatofibroma were HHV8 positive, whereas two of the atypical vascular lesions were viral positive, leading to a final diagnosis of Kaposi's sarcoma.

Conclusions: RT in situ PCR for HHV8 RNA is a useful tool in the differential diagnosis between Kaposi's sarcoma and its mimics.
A possible role of catenin dyslocalization in pemphigus vulgaris pathogenesis
Authors: Muzio Lorenzo Lo , Pannone Giuseppe , Staibano Stefania , Mignogna Michele Davide , Rubini Corrado , Ruocco Eleonora , De Rosa Gaetano , Sciubba James J.
Published: March 2003

Background: Pemphigus vulgaris (PV) is an autoimmune blistering disease of the skin and mucosa due to the presence of autoantibodies against the components of desmosomes. To date, less is known about the expression levels of β‐ and γ‐catenins in blistering diseases. The objective of this study was to evaluate the role of β‐ and γ‐catenins in the pathogenesis of acantholysis in pemphigus vulgaris.

Methods: β‐ and γ‐catenin expression was evaluated by immunohistochemistry in 30 cases of PV at various stages of the disease and, as controls, in 18 specimens of the skin/oral mucosa of healthy patients.

Results: Healthy skin and normal oral mucosa showed a strong β‐ and γ‐catenin expression in basal and spinous layers with a prevalent cellular membrane distribution; the intensity of staining progressively decreased toward the superficial layers of epithelium. In PV patients, cytoplasmic expression of γ‐catenin was detected in 28/30 cases, and in 19/30 cases of PV for β‐catenin. Moreover, a progressive displacement of the signal toward the nucleus was found in 14/30 cases for β‐catenin, with dyslocalization toward the nucleus, particularly in areas with intense acantholysis, and in 22/30 cases of PV for γ‐catenin.

Conclusions: Abnormal distribution of γ‐catenin, consequent to PV IgG, may be considered a direct consequence of Dg3 dissociation from catenin. γ‐catenin likely plays a direct role in PV pathogenesis through its dyslocalization toward the nucleus or indirectly through the β‐catenin dyslocalization toward the nucleus, which is thought to induce transcription of selected target genes, such as uPAR.
Intrauterine epidermal necrosis
Authors: Allee Julie E. , Saria Elizabeth A. , Rosenblum Dennis , Prose Neil S. , Prieto Victor G. , Shea Christopher R.
Published: March 2003

Background: Epidermal necrosis in a neonate is an uncommon event with a variety of potential cases.

Result: We report a case of intrauterine epidermal necrosis in a preterm infant, with death occurring soon after birth. The histopathology of the denuded skin revealed full‐thickness epidermal necrosis and calcification within both the epidermis and follicular structures.

Conclusion: We believe this represents the fourth reported case of lethal intrauterine epidermal necrosis and follicular calcification.
Specific cutaneous infiltrates of B‐cell chronic lymphocytic leukemia (B‐CLL) at sites typical for Borrelia burgdorferi infection
Authors: Cerroni Lorenzo , Höfler Gerald , Bäck Barbara , Wolf Peter , Maier Gerlinde , Kerl Helmut
Published: March 2003

Background: Cutaneous manifestations of B‐cell chronic lymphocytic leukemia (B‐CLL) comprise a wide spectrum of clinicopathologic presentations. In some cases, onset of skin lesions is triggered by antigenic stimulation, and specific skin infiltrates at sites of previous herpes simplex or herpes zoster infection have been well documented. Specific skin manifestations of B‐CLL can also be observed at sites typical for lymphadenosis benigna cutis (nipple, scrotum, earlobe), a Borrelia burgdorferi‐associated cutaneous B‐cell pseudolymphoma.

Methods:  We studied specific skin manifestations of B‐CLL arising at sites typical for B. burgdorferi‐induced lymphadenosis benigna cutis, analyzing tissues for presence of B. burgdorferi DNA using the polymerase chain reaction (PCR) technique. Six patients with B‐CLL (M : F = 4 : 2; mean age: 67.8) presented with specific skin lesions located on the nipple (four cases) and scrotum (two cases).

Results:  Clinically there were solitary erythematous plaques or nodules. Histology revealed in all cases a dense, monomorphous infiltrate of small lymphocytes showing an aberrant CD20+/CD43+ phenotype. In all cases monoclonality was demonstrated by PCR analysis of the JH gene rearrangement. PCR analysis showed in four of the six cases the presence of DNA sequences specific for B.burgdorferi.

Conclusions:  Our study demonstrates that infection with B. burgdorferi can trigger the development of specific cutaneous infiltrates in patients with B‐CLL.
Host‐vs.‐altered‐host eruptions in patients on liposomal doxorubicin
Authors: Skelton Henry , Linstrum Jill , Smith Kathleen
Published: March 2003

Background:  Anthracycline antitumor antibiotics are the most commonly used chemotherapeutic agents. One of these is doxorubicin. Liposomal doxorubicin was developed as a drug delivery system in order to deliver the active drug intracellularly while decreasing the systemic toxicity, particularly hematological and cardiac toxicity.

Methods:  The clinical and histologic findings of the cutaneous eruptions of associated with liposomal doxorubicin are reviewed.

Results: The eruptions occurred in three women with metastatic ovarian carcinoma who were treated with liposomal doxorubicin. These three patients developed erythematous macular/papular to plaque cutaneous lesions, and in one patient a vesicular component. The eruptions involved the trunk and extremities approximately 3–4 weeks after completions of therapy. None of the patients had any documented infections, and none of the patients had symptoms other than pruritus. The eruptions cleared over a period of weeks to months. Histologic features included an interface dermatitis with numerous apoptotic/dyskeratotic cells within the epidermis, with involvement the intra‐epidermal sweat ducts and the infundibulum of hair follicles.

Conclusion:  We believe that these eruptions represented a chemotherapy induced host‐vs.‐altered host reaction.
Anaplastic neoplasms arising from basal cellcarcinoma xenotransplants into SCID‐beige mice
Authors: Carlson J. Andrew , Combates Nicholas J. , Stenn Kurt S. , Prouty Stephen M.
Published: March 2003

Background: An animal model for the study of basal cell carcinoma (BCC) is required to better understand its biology. Several attempts to grow BCC in immuno‐incompetent animals have been only modestly successful.

Methods: To test the ability of BCC to grow in a mouse with complete and severe immuno‐incompetence, 14 individual BCC were transplanted into the subcutaneous tissue of 18 SCID‐beige mice (T, B and natural killer cell deficient). Light microscopy and immunophenotypic analyses were performed on primary BCC and first and seventh passage tumors.

Results: Transplantation of three BCC yielded rapidly growing anaplastic tumors for a tumor take of 18% (3/18). SCID‐beige mice without tumor growth had mostly scars or epidermoid cysts at the transplant sites. The three patients whose BCC gave rise to the anaplastic tumors were significantly older than those without tumor growth (87 vs. 64, p = 0.001), but they did not differ with respect to BCC type or general health. These three anaplastic tumors were histologically and immunophenotypically similar, being composed of dyscohesive, pleomorphic cells that expressed vimentin and smooth muscle actin. In the first passage mice these tumors were locally invasive, tumor‐forming nodules associated with an expansion of donor inflammatory cells (T and B lymphocytes and plasma cells), rare remnants of BCC epithelium and epidermoid cysts. By the seventh passage, the tumors were homogenous and metastasized widely throughout the mice. Changing transplantation location to the dermis to wound environment or supplementing the tumor with BCC‐derived fibroblasts did not alter the phenotype or growth rate in SCID‐beige mice. Anaplastic tumors also grew easily in SCID mice (T and B cell deficient). However, transplantation of the anaplastic tumors into normal mice (CB‐17) or less severely immunodeficient mice (NCr and Balb/c: T and natural killer cell deficient) did not allow for growth. Furthermore, tumor growth could not be maintained in vitro.

Conclusion: Empirically, these data suggest that BCC has the potential to become an aggressive metastatic neoplasm, given the right immune and stromal environment. Moreover, a functional B lymphocyte system appears to prevent this growth. As human lymphocytes also engraft in SCID‐beige mice, the original host immune response could be responsible for the lack of tumor growth in the majority of xenografts. Furthermore, the anaplastic and metastatic phenotype of these BCC derived neoplasms may be the experimental equivalent of metastatic BCC and BCC associated with carcinosarcoma.
A predominant IgG4 subclass may be responsible for false‐negative direct immunofluorescence in bullous pemphigoid
Authors: Buschman Kerry E. , Seraly Mark , Thong H. Y. , Deng Jau‐Shyong , Draviam Rose P. , Abernethy John L.
Published: March 2003

Background: Bullous pemphigoid (BP) is an immune‐mediated blistering disease, usually characterized immunopathologically by the linear deposition of IgG and C3 along the basement membrane zone (BMZ) of skin. However, positive deposition of C3 but negative staining for IgG on direct immunofluorescence (DIF) studies has been noted in some patients.

Methods: Twelve patients known to have BP but with absence of staining for IgG were included in this study. Frozen sections of skin specimens from the 12 patients were subjected to IgG DIF, as well as a sandwich double antibody method of staining for IgG, IgG subclasses, and light chains. Enzyme‐linked immunosorbent assay (ELISA) using commercially available human IgG subclasses was used to analyze the subclass restriction of FITC‐labeled antihuman IgG conjugates.

Results:  Of the 12 skin specimens with positive C3 and negative IgG on DIF, nine were positive for IgG with the double antibody sandwich method. In addition, all 12 specimens had positive linear staining for the subclass IgG4 along the BMZ with this method. There was no IgG light chain restriction. Two commercially obtained antihuman IgG conjugates, both commonly used in our laboratory for DIF testing, were analyzed for separate IgG subclass specificity by ELISA. Both conjugates showed high reactivity to IgG1 and IgG3 with less reactivity to IgG2 and IgG4.

Conclusion:  These results suggest that the following factors contribute to false‐negative staining for IgG on DIF in some BP patients: (i): subthreshold IgG in skin specimens; (ii) limited reactivity of commercial antihuman IgG conjugates to the IgG4 subclass; and (iii) decreased sensitivity of DIF compared with double antibody methods for the detection of IgG. The use of sandwich double antibody immunofluorescence methods to test for IgG and/or IgG subclasses may be helpful in definitively diagnosing BP in patients with negative IgG and positive C3 staining on DIF.
Human telomerase RNA component expression in Spitz nevi, common melanocytic nevi, and malignant melanomas
Authors: Guttman‐Yassky Emma , Bergman Reuven , Manov Lena , Sprecher Eli , Shaefer Yan , Kerner Hedviga
Published: March 2003

Background:  Telomerase is a ribonucleoprotein DNA polymerase that is capable of synthesizing telomeres onto the ends of chromosomes. The cumulative loss of telomerase activity is believed to be associated with cell senescence. Telomerase activity has been shown to be higher in malignant melanomas than in common melanocytic nevi. The aim of the present study was to elucidate the pattern of expression of the human telomerase RNA (hTER) component in routinely processed specimens of Spitz nevi, malignant melanomas, and ordinary melanocytic nevi.

Methods:  Ten specimens of each type of tumor were studied, using an in situ hybridization technique.

Results:  All three types of tumors demonstrated moderate to high intensities of hTER expression, usually in more than half of the tumor cells, and the majority of the studied lesions in each group did not show stratification of staining. The hTER component was also detected in the epidermis, sweat glands, and pilosebaceous units.

Conclusions:  hTER levels do not necessarily correlate with the level of telomerase activity, and the level and pattern of hTER expression are not useful as an adjunct to the histologic differential diagnosis of Spitz nevi from melanocytic nevi and malignant melanomas.
h‐Caldesmon as a specific marker of smooth muscle cell differentiation in some soft tissue tumors of the skin
Authors: D'Addario Stephen F. , Morgan Michael , Talley Lori , Smoller Bruce R.
Published: March 2003

Background:  An existing problem in contemporary pathology is the classification and distinction of spindle cell soft tissue tumors of the skin. Markers such as α‐smooth muscle actin (α‐SMA) and desmin, considered specific for smooth muscle cell (SMC), have been shown to be expressed in a variety of fibroblastic and myofibroblastic processes. High‐molecular‐weight caldesmon (h‐caldesmon), one of two isoforms, is reported to be expressed exclusively by SMC and has recently been shown to be a specific marker of SMC tumors.

Methods: Tumors were obtained from 11 patients taken from the surgical pathology archives of the University of South Florida and cases were coded as smooth muscle hamartoma, myofibroma, and dermatomyofibroma.

Results:  The case of smooth muscle hamartoma had greater than 90% of tumor cells labeling with anti‐h‐caldesmon antibodies. Three of three cases of myofibroma had focal areas of positivity representing less than 10% of total tumor cells. Seven of seven dermatomyofibromas showed no apparent labeling with anti‐h‐caldesmon antibody. Dense reactivity was noted in vascular wall smooth muscle, indicating internal controls.

Conclusions: We can conclude that h‐caldesmon is a specific marker of fully differentiated smooth muscle and that it can serve to differentiate spindled SMC soft tissue tumors of the skin from tumors of myofibroblastic and/or fibroblastic origin.
Three‐dimensional imaging of human skin and mucosa by two‐photon laser scanning microscopy
Authors: Malone Janine C. , Hood Antoinette F. , Conley Tameka , Nürnberger Jens , Baldridge Lee Ann , Clendenon Jeffrey L. , Dunn Ken W. , Phillips Carrie L.
Published: March 2003

Background: Various structural components of human skin biopsy specimens are difficult to visualize using conventional histologic approaches.

Methods: We used two‐photon microscopy and advanced imaging software to render three‐dimensional (3D) images of in situ nerves, blood vessels, and hair follicles labeled with various fluorescent markers. Archived frozen human skin biopsy specimens were cryosectioned up to 150 µm in thickness and fluorescently stained with rhodamine‐ or fluorescein‐labeled antibodies or lectins. Optical sections were collected by two‐photon microscopy and the resulting data sets were analyzed in three dimensions using Voxx software.

Results: Reconstructed image volumes demonstrated the complex 3D morphology of nerves, blood vessels and adnexal structures in normal mucocutaneous tissue.

Conclusion: Two‐photon microscopy and Voxx rendering software allow for detailed 3D visualization of structures within human mucocutaneous biopsy specimens, as they appear in situ, and facilitate objective interpretation of variations in their morphology. These techniques may be used to investigate disorders involving cutaneous structures that are difficult to visualize by means of traditional microscopy.
Aberrant expression of p16 and RB protein in eccrine porocarcinoma
Authors: Gu Li‐Hong , Ichiki Yoshiro , Kitajima Yasuo
Published: March 2003

Background:  Eccrine porocarcinoma is a rare malignant tumor of the sweat gland and very little is known about its etiology and molecular basis.

Methods:  To investigate the expression of p16 and retinoblastoma (RB) protein and their relationship, an immunohistochemical method was performed on nine eccrine porocarcinomas and five eccrine poromas. Furthermore, one case of eccrine porocarcinoma was analyzed for p16 gene mutation.

Results:  A striking inverse correlation between p16 and RB expression was noted in all of the eccrine porocarcinomas and poromas. Strong immunoreactivity for p16 protein was observed in both nuclei and cytoplasm of the tumor cells in eight out of nine cases of eccrine porocarcinomas, while RB expression was negative in these cases. Conversely, one case of eccrine porocarcinoma did not show immunoreactivity for p16 protein, whereas RB protein was positive in the scattered nuclei. On the other hand, immunostaining of p16 was negative in all cases of five poromas, whereas RB‐positive nuclei were sparse. No p16 gene mutation was detected in the investigated eccrine porocarcinoma case.

Conclusions:  These results indicate that detectable p16 protein and loss of RB protein are common occurrences in eccrine porocarcinoma lesions. Moreover, overexpression of p16 protein may be an additional, simple and useful diagnostic marker for eccrine porocarcinoma on routine laboratory screening.
The value of extended‐setting‐time amalgam in pre‐clinical practice in undergraduate dental training
Authors: Jedynakiewicz N. M. , Urbanska D. K.
Published: March 2003

This is a single‐blind controlled study investigating the advantages of extended‐setting time dental amalgam compared to a standard amalgam in the training of undergraduate dental students. 50 dental students were randomly assigned 1 of the 2 alloys for each restoration placed in the study. Each student placed class I and class II restorations in extracted human teeth mounted in typodont arches. 449 completed restorations were available for assessment, 228 restorations in the experimental group and 221 restorations in the control group. The test restorations were overall marginally inferior to those of the control group. There appears to be no advantage in using extended‐setting time dental amalgam in undergraduate training.
Teaching and assessing behavioural techniques of applied relaxation for reduction of dental fear using a controlled chairside simulation model
Authors: McGoldrick P. M. , Pine C. M.
Published: March 2003

This paper describes a novel method of teaching and assessing relaxation techniques using a controlled chairside simulation model. 50 pre‐clinical dental undergraduates provided an evaluation of the teaching model as part of their basic clinical skills teaching. Students were required to demonstrate applied relaxation techniques according to 2 pre‐selected clinical vignettes. Inter‐observer reliability of assessing student competence was calculated on a varied sample (n=9) using 2 independent observers and found to be adequate. The majority of the students viewed the teaching and assessment method as interesting and clinically relevant. However, a few students reported that the method was less advantageous than using a real patient. It is concluded that using vignettes as part of clinical training accompanied by video‐based feedback provides a resourceful learning tool for pre‐clinical teaching in the absence of patient contact.
The repair of direct composite restorations: an international survey of the teaching of operative techniques and materials
Authors: Blum I. R. , Schriever A. , Heidemann D. , Mjör I. A. , Wilson N. H. F.
Published: March 2003

This article reports the findings of a study on the aspects of the teaching of repair as a conservative alternative to the replacement of failing direct composite restorations (DCRs) in primary dental degree curricula in the UK and Ireland, Germany and Scandinavia. Data on teaching, including operative techniques applied in the repair of DCRs, were collected by means of a questionnaire sent to 58 dental schools in the surveyed countries. Based on the overall response rate of 83%, the findings indicate that the majority (59%) of the schools in the countries surveyed may be found to teach the repair of DCRs. However, marked variations were observed in respect of this teaching and the expected longevity of repaired DCRs. Where the repair of DCRs was not taught, views differed as to the intentions, if any, to include this teaching in the curriculum. It is concluded that the teaching of DCRs may be found to be widespread in dental schools in the UK and Ireland, Germany and Scandinavia, with this teaching being subjected to considerable variation within and between countries.
Beetle species richness along the forest productivity gradient in northern Finland
Authors: Similä Maarit , Kouki Jari , Mönkkönen Mikko , Sippola Anna‐Liisa
Published: March 2003

The occurrence and habitat associations of the majority of invertebrate groups in boreal forests are poorly known, even though these groups represent perhaps over 99% of the animal species diversity in the forests. We studied the beetle (Coleoptera) fauna of four forest site types in northern Finland: in spruce mires, herb rich, mesic and sub‐xeric forests. We sampled beetles in 32 study sites with five window and five pitfall traps in each. We describe the species abundance and diversity patterns within and among forest types and relate these patterns to structural components of the forests. The volume of decaying wood varied from 14 to 93 m3 ha−1 among sampling sites. The total beetle catch consisted of 100 333 individuals and 435 species. The beetle species richness did not vary according to site fertility but the number of specimens increased with increasing fertility in heath forest sites. The richness of beetle species correlated only weakly with any of the stand structure characteristics at the stand level. Nevertheless, the detrended correspondence analysis (DCA) indicated that different beetle assemblages are characteristic of different forest types. The high level of beta‐diversity in beetles among forest types indicates that focusing exclusively on, for example, key‐biotopes (presumed biodiversity hotspots) when selecting areas to be set aside would result in a situation where a large proportion of species, even of the rare and threatened ones, is not included in this network of protected areas. This suggests that the complementary set of different forest types may be the best general strategy to maintain the overall beetle species diversity in boreal forests.
Size‐dependent species‐area relationships in benthos: is the world more diverse for microbes?
Authors: Azovsky Andrey I.
Published: March 2003

Using original and literature data on species richness, I compared the species‐area relations for 5 different size classes of the Arctic benthos: macrofauna sensu lato, polychaetes, nematodes, ciliates and diatom algae. The data pool covered a wide range of areas from single samples to the whole seas. Both the slopes and intercepts of the curves depended significantly on the logarithm of the mean body size of the group. The number of small species (ciliates and diatom algae) showed relatively higher local diversity but increased more slowly with the area than the number of larger ones. Thus, both α‐ and β‐components of species diversity of the marine benthos were size‐dependent. As a consequence, the actual relations between number of species and their physical size are spatially scale‐dependent: there are many more species of smaller size classes in any one local community, but at a global scope the situation changes drastically. The possible reasons are discussed, including dispersal efficiency, rates of speciation and size‐dependent perception of environmental heterogeneity. Body size is suggested to be the important scaling factor in manifestation of so‐called “general ecological laws”.
Increased serum IgA and decreased IgG3 strongly correlate with increased serum TGF‐β1 levels in patients with nonimmune chronic idiopathic neutropenia of adults
Authors: Papadaki Helen A. , Palmblad Jan , Kapsimali Violetta , Anagnou Nikolas P. , Eliopoulos George D.
Published: March 2003

Abstract: Objective: To study the changes in serum immunoglobulins and some closely related pro‐inflammatory cytokines in patients with nonimmune chronic idiopathic neutropenia of adults (NI‐CINA). Methods: Serum levels of gamma‐globulins, IgG, IgA, IgM, IgG subclasses, interleukin‐4 (IL‐4), interferon‐gamma (IFN‐γ) and transforming growth factor‐beta1 (TGF‐β1) were evaluated in 83 NI‐CINA patients and 65 normal controls using the respective conventional methods. Results: We found that serum gamma‐globulin, IgG and IgG1 levels were all significantly increased in the entire group of patients studied, compared to controls (p<0.001, p<0.01 and p<0.01, respectively), while the levels of IgG3 were significantly reduced (p<0.001). Serum IgA were increased in patients with severe neutropenia (p<0.001). No significant changes were noted in serum IgM, IgG2 and IgG4 levels. The infrequent occurrence of detectable amounts of IL‐4 and IFN‐γ in the serum was similar in both, patients and control subjects. Serum levels of TGF‐β1 were increased in all groups of patients studied and they correlated inversely with the levels of IgG3 (p<0.001) and positively with the levels of IgA (p<0.001), suggesting the possible involvement of the cytokine in immunoglobulin class switching. Conclusion: Patients with NI‐CINA have significant changes in serum immunoglobulins and some inflammation‐related cytokines. These findings provide additional evidence for the existence of an unrecognized low‐grade chronic inflammatory process in NI‐CINA patients and coroborate our previously reported suggestion for the possible involvement of this inflammation in the pathogenesis of neutropenia in the affected subjects.
Real‐time quantitation of bcr‐abl transcripts in haematological malignancies
Authors: Saffroy R. , Lemoine A. , Brezillon P. , Frenoy N. , Delmas B. , Goldschmidt E. , Souleau B. , Nedellec G. , Debuire B.
Published: March 2003

Abstract: We have applied an automated real‐time quantitative PCR assay using a double‐labeled fluorogenic probe to detect t(9;22)‐positive cells in haematological malignancies. The results are expressed as the ratio of chimeric bcr‐abl transcripts on abl transcripts. Highly reproducible results were obtained for t(9;22)‐positive K562 RNA. Ten copies of bcr‐abl DNA from a recombinant KW‐3 plasmid and one positive cell in 104 can be detected. Thirty‐two patients with chronic myeloid leukaemia (CML), 25 with acute leukaemia, 12 with myelodysplastic syndromes and 7 with other myeloproliferative syndromes were tested. Follow‐up data were obtained in bcr‐abl positive cases. Results were compared with those of conventional nested RT‐PCR and cytogenetics. Real‐time quantitative RT‐PCR values correlated well with both these methods. However, in some cases the only means of detecting early relapse or blastic transformation was to examine the kinetics of real‐time quantitative RT‐PCR. Thus, real‐time quantitative RT‐PCR appears suitable for the diagnosis and follow‐up of patients with the t(9;22) translocation.
Treatment outcome in patients younger than 60 years with advanced stages (IIB–IV) of Hodgkin's disease: the Swedish National Health Care Programme experience
Authors: Amini Rose‐Marie , Enblad Gunilla , Gustavsson Anita , Ekman Tor , Erlanson Martin , Haapaniemi Eva , Glimelius Bengt
Published: March 2003

Abstract: Background –Despite improved treatment results achieved in Hodgkin's disease (HD), only about 70% of patients with advanced stages are cured. The primary aim of this study was to evaluate the outcome of advanced stages (IIB–IVB) of HD in younger patients in an unselected population‐based group of patients. The patients were recommended individualized treatment with respect to number of chemotherapy (CT) courses and post‐CT radiotherapy (RT) based on pretreatment characteristics and tumour response. Secondly, we investigated if variables of prognostic importance could be detected. Patients and methods –Between 1985–92, 307 patients between 17–59 yr of age (median 36) were diagnosed with HD in stages IIB–IVB in 5/6 health care regions in Sweden. Median follow‐up time was 7.8 yr (1.3–13). Retrospectively, laboratory parameters were collected. Results –In total, 267 (87%) patients had a complete response (CR). The overall and disease‐free 10‐yr survivals in the whole cohort were 76% and 67%, respectively. There was no difference in survival between the groups of patients who received 6 or 8 cycles of CT. Survival was not higher for patients in CR after CT when RT was added. For those in PR after CT, additional RT raised the frequencies of CR. A selected group of pathologically staged patients was successfully treated with a short course (2 cycles) of CT+RT. In univariate analyses survival was affected by age, stage IVB, bone‐marrow involvement, B‐symptoms, S‐LDH, S‐Alb and reaching CR or not after 2, 4 and 6 cycles of CT. In a multivariate analysis, age and reaching CR after 6 cycles of CT remained statistically significant. Conclusions –The lack of difference in survival between the groups of patients who received 6 versus 8 cycles of CT indicates a successful selection of patients for the shorter treatment. Reaching a rapid CR significantly affected outcome. Whether some patients need less CT than the generally recommended 8 courses can properly only be evaluated in a randomised study. Additional RT may play a role in successful outcome, particularly if residual tumours are present, but its precise role can also only be defined in prospectively randomised studies. Reaching CR after CT was the most important variable affecting survival besides age.
Deafness from eighth cranial nerve involvement in a patient with large‐cell transformation of mycosis fungoides
Authors: Chua Susan L. , Seymour John F. , Prince H. Miles
Published: March 2003

Abstract: Central nervous system (CNS) involvement by mycosis fungoides (MF) is rare and is usually seen in advanced stages with lymph node or visceral involvement. We describe a patient with advanced stage MF in large‐cell transformation who presented with profound hearing loss after chemotherapy. Despite an initial differential diagnosis of vincristine‐related neurotoxicity based on clinical, audiometric, and MRI investigations, CSF examination revealed lymphomatous leptomeningeal involvement. This case illustrates the importance of an awareness of the possibility of CNS involvement by MF and underlines the need for a complete neurologic evaluation including CSF examination in a patient with underlying MF who presents with a new neurological problem.
Transfusion‐dependent congenital dyserythropoietic anaemia with intraerythroblastic inclusionsof a non‐globin protein
Authors: Iolascon A. , Martire B. , Lee M. J. , Wickramasinghe S. N.
Published: March 2003

Abstract: The congenital forms of dyserythropoiesis comprise a group of hereditary disorders characterized by ineffective erythropoiesis as the predominant mechanism of anaemia and morphologically abnormal erythroblasts. Up to now three major forms and four variants have been described. Group VII is characterized by dyserythropoiesis with intraerythroblastic precipitation of a non‐globin protein. Here we described a case of dyserythropoietic anaemia presenting neonatally and requiring regular blood transfusions. Optical and electronic microscopy studies confirmed that this case was very similar to those in two previously reported transfusion‐dependent patients with an unusual type of congenital dyserythropoietic anaemia.
Investigation for the presence of anti‐erythropoietin antibodies in patients with myelodysplastic syndromes
Authors: Voulgari Paraskevi V. , Hatzimichael Eleftheria ChR. , Tsiara Stavroula , Tzallas Christos , Drosos Alexandros A. , Bourantas Konstantinos L.
Published: March 2003

Abstract: Objectives: Recombinant human erythropoietin (rHuEpo) improves anemia in 25% of patients with myelodysplastic syndromes (MDS). The variable and sometimes low response rate to rHuEpo treatment raises the question whether the existence of autoantibodies against erythropoietin (epo) is partially responsible. In the present study we investigated the presence of anti‐epo autoantibodies in MDS patients. Methods: Forty‐three patients with MDS were studied. Sixteen patients had refractory anemia (RA), 13 had RA with ringed sideroblasts, 3 had RA with excess of blasts (RAEB), 9 had RAEB in transformation and 2 patients had chronic myelomonocytic leukemia. They were divided in 3 groups according to rHuEpo treatment. Group A consisted of 10 patients who did not receive rHuEpo treatment. Group B included 13 patients who were on rHuEpo treatment (150 IU/kg subcutaneously, 3 times weekly) showing an increase of hemoglobin (Hb) values or reduction of transfusion requirements and Group C consisted of 20 patients who did not respond or stopped responding to rHuEpo treatment. Laboratory studies consisted of a complete blood cell count, measurement of serum epo and determination of anti‐epo antibodies using ELISA. Results: There were no significant differences with regard to age and sex among the three groups. No autoantibodies against epo were found in the examined sera, apart from a female patient from group A who showed a low positive titer. Conclusion: We suggest that anti‐epo autoantibodies do not contribute to the development of MDS‐related anemia and are not responsible for the modest response to rHuEpo treatment. Further investigation is needed to identify possible reasons for the low response rate to rHuEpo treatment.
Mobilisation of tumour cells along with CD34+cells to peripheral blood in multiple myeloma
Authors: Knudsen Lene Meldgaard , Rasmussen Thomas , Nikolaisen Kirsten , Johnsen Hans Erik
Published: March 2003

Abstract:Background:Cells belonging to the malignant clone are found in the peripheral blood in myeloma patients. In order to minimise the content of tumour cells in the stem cell product it is crucial to perform stem cell harvest at a time when tumour cells in the peripheral blood are at a minimum.Objective:The aim of the study was to compare the mobilisation kinetics of normal CD34+cells and myeloma plasma cells during mobilisation with either G‐CSF alone or high‐dose cyclophosphamide (HDCy) plus G‐CSF.Design and methods:Morning blood samples were drawn each day during mobilisation from start of G‐CSF or HDCy and to the end of leukapheresis, and were analysed by flow cytometry for content of CD34+cells and myeloma plasma cells (CD38+ + CD45−). Tumour cells were also estimated by a patient‐specific real‐time polymerase chain reaction (PCR) method based on the 5′ nuclease TaqMan technology.Results:Flow cytometry data from 16 patients showed concomitant mobilisation of CD34+cells and myeloma plasma cells. Seven patients were mobilised twice; first with G‐CSF alone and then with HDCy plus G‐CSF. There was no difference between the two mobilisation regimens regarding tumour cell mobilisation kinetics. Real‐time PCR was performed in one patient and confirmed the mobilisation of tumour cells at the time when CD34+blood cells were at a maximum.Conclusions:Tumour cells are mobilised to the peripheral blood at the same time as CD34+cells in multiple myeloma patients after priming with both G‐CSF alone and HDCy in combination with G‐CSF.
Hydroxyurea and periodicity in myeloproliferative disease
Authors: Bennett Michael , Grunwald Arthur J.
Published: March 2003

Abstract: Three patients, one with polycythaemia vera (PV) and two with chronic myeloid leukaemia (CML), are described who had cycling of blood counts which became apparent whilst receiving hydroxyurea therapy. Significant periodicity was confirmed with the use of the Lomb periodogram. This is Fourier power spectral analysis tailored for unevenly sampled data. The patient with PV had marked oscillations of platelet counts with a periodicity of 29 d and an amplitude of (202–588)×109/L. Smaller oscillations of neutrophil, monocyte and lymphocyte numbers and Hb levels occurred with a similar periodicity. Anticipatory changes in hydroxyurea dosage or the maintenance of a constant dose did not abolish periodicity, but a change in therapy to the non‐cycle‐specific drug anagrelide dampened and abolished the cycling. One of the patients with CML had tremendous and clear oscillations in white cell, platelet and Hb levels, with a mean periodicity of 74 d. The other had erratic counts which were confirmed to be significantly periodic (64 d), on spectral analysis. A change in therapy to busulphan in both these patients again dampened and abolished the cycling. Hydroxyurea, which is a cell‐cycle‐specific agent, probably exacerbates the periodicity which may be present in some patients with myeloproliferative disease. A change in therapy to non‐cycle‐acting compounds such as busulphan or anagrelide results in much more stable counts in such patients.
Sickle erythrocytes increase prostacyclin and endothelin‐1 production by cultured human endothelial cells under flow conditions
Authors: Shiu Yan‐Ting , McIntire Larry V. , Udden Mark M.
Published: March 2003

Abstract: We investigated the effects of sickle erythrocytes on the production of vasotone mediators in endothelial cells (ECs) using an in vitro recirculating flow system. Sickle erythrocytes increased the EC production of two important vasoactivators, prostacyclin and endothelin‐1, under venous wall shear stress conditions of 1 dyn cm−2. The presence of interleukin‐1β in the perfusion system, as a model for inflammatory cytokine effects, enhanced the overall amounts of released prostacyclin but did not affect the production of endothelin‐1. This study demonstrates the effects of sickle erythrocytes on the function and metabolism of ECs under vascular flow environments. The altered production of vasoactivators may contribute to the vasotone instability and vasoocclusive crises in sickle cell anemia.
Serum erythropoietin (EPO) levels correlate with survival and independently predict response to EPO treatment in patients with myelodysplastic syndromes
Authors: Wallvik Jonas , Stenke Leif , Bernell Per , Nordahl Gunnar , Hippe Erik , Hast Robert
Published: March 2003

Abstract: Treatment with recombinant erythropoietin (EPO) can alleviate anaemia in patients with myelodysplastic syndromes (MDS). The present study, based on a long‐term follow‐up of 68 MDS patients (26RA, 16 RAS, 26 RAEB) treated with EPO alone, pinpoints pre‐treatment variables associated with response induction, response duration and overall survival. Response, defined as an increase in haemoglobin >15 g L−1 or eliminated erythrocyte transfusion requirements, was observed in 22 of 66 (33%) evaluable patients. The median response duration was 15 (range 3–64 +) months. Using univariate logistic regression models, responders displayed significantly lower baseline serum EPO levels (S‐EPO), more often normal bone marrow blast cell content (RA/RAS vs. RAEB), normal cytogenetics and no need for erythrocyte transfusion. In a multiple logistic regression model, S‐EPO (P = 0.009), marrow blast content (P = 0.031) and erythrocyte transfusion need (P = 0.024) remained associated with response induction. The probability of response for a patient with S‐EPO >50 U L−1, RA/RAS and no transfusion need was 0.79 (0.53–0.93, 95% CI). The median overall survival time from start of EPO treatment was 26 months, significantly longer for responders than for non‐responders (49 vs. 18 months, P = 0.018). Survival was also predicted by baseline S‐EPO; patients with S‐EPO > 50 U L−1 (n = 50) had a median survival of 17 months, as compared to 65 months for those with S‐EPO >50 U L−1 (n = 14, P = 0.024). The international prognostic scoring system (IPSS) for MDS predicted survival (P = 0.003) and progression to acute leukemia (P < 0.001) but not response to EPO treatment. Furthermore, in a logistic regression model with S‐EPO and IPSS, S‐EPO (but not IPSS) was again a significant predictor for response (P = 0.007). Our data facilitate the optimal selection of MDS patients suitable for EPO treatment and pinpoint S‐EPO as a powerful predictor of response and overall survival in MDS.
Paraneoplastic cerebellar degeneration and nephrotic syndrome preceding Hodgkin's disease: case report and review of the literature
Authors: Spyridonidis A. , Fischer K.‐G. , Glocker F. X. , Fetscher S. , Klisch J. , Behringer D.
Published: March 2003

Abstract: A patient presented with symptoms of cerebellar degeneration and nephrotic syndrome. A work‐up at that time failed to reveal an underlying disease; however, 20 months later Hodgkin's disease was diagnosed. Hodgkin's lymphadenopathy developed 2 wk after prednisone therapy for the nephrotic syndrome had been discontinued. Systemic polychemotherapy resulted in complete remission of both Hodgkin's disease and nephrotic syndrome, while the neurological deficit persisted. Patients with unexplained cerebellar degeneration and/or nephrotic syndrome demand extensive evaluation for the presence of Hodgkin's disease, and steroid therapy may delay diagnosis.
Prolymphocytic leukaemia and Hodgkin's lymphoma
Authors: Naranjo Tomás Álvaro , Matutes Estel·la , Martínez Joaquín Jaén , Font Ferré Llorenç , Domingo Alicia , Príncep Ramón Bosch , Usach Teresa Salvadó
Published: March 2003

Abstract: We describe a case of B‐prolymphocytic leukaemia (B‐PLL) who, following a long‐lasting remission with fluradabine, developed a Hodgkin's lymphoma (HL) with bone marrow involvement. A 75‐yr‐old male was found to have a lymphocytosis [white blood cell (WBC) count = 146 × 109 L−1], small volume axillary lymphadenopathy, and hepatosplenomegaly. The majority of circulating lymphocytes had a round nucleus and prominent single nucleolus. The patient did not respond to chlorambucil and then received fludarabine, achieving a good response lasting for 5 yr, when he manifested with B symptoms and pancytopenia. A diagnosis of HL in the bone marrow was made based on histology (Reed–Sternberg cells) and immunohistochemistry (CD30+, CD15+). Epstein–Barr virus (EBV) studies were negative. The patient was treated with chemotherapy but died 6 months later from disease progression.
Simultaneous development of lymphoplasmacytic lymphoma and diffuse large B‐cell lymphoma – analyses of the clonal relatedness by sequencing CDR3 in immunoglobulin heavy chain genes
Authors: Shimizu Seiichi , Tamagawa Yuna , Kojima Hiroshi , Mori Naoyoshi , Nagata Michio , Noguchi Masayuki , Nagasawa Toshiro
Published: March 2003

Abstract: A 75‐yr‐old male simultaneously having lymphoplasmacytic lymphoma (LPL) and diffuse large B‐cell lymphoma (DLBCL) is presented. He had manifested with lumbago, high‐grade fever, and confusion. Physical examination on admission showed disorientation and severe back pain. There were neither lymphadenopathy nor hepatosplenomegaly. Routine laboratory tests showed moderate pancytopenia, hypercalcemia (serum calcium, 15.9 mg/dL), IgM λ‐type monoclonal gammopathy (IgG, 405 mg/dL; IgA, 42 mg/dL; and IgM, 2023 mg/dL), and λ‐type Bence–Jones protein in the urine (0.8 g/d). Bone marrow biopsy showed the clusters of surface λ‐positive small‐sized mature‐appearing lymphoplasmacytoid cells. Bone survey and computed tomographic scan showed multiple osteolytic lesions and a tumor involving the third lumbar spine (L3). An open biopsy of the L3 tumor showed diffuse proliferation of CD20‐ and λ‐positive large cells. We thus diagnosed the patient as simultaneously having LPL and DLBCL. Although the combination chemotherapy was at least partially effective, he died of bacteremia and organ failure after three courses of chemotherapy. To clarify the clonal relatedness between LPL and DLBCL, we analyzed the sequences of the complementarity‐determining region 3 in immunoglobulin heavy‐chain genes. The data showed that LPL and DLBCL in the present patient originated from two independent clones.
Regulation of cholesterol synthesis by oleic and palmitic acid in keratinocytes
Authors: Siefken W. , Höppner H. , Harris I. R.
Published: March 2003

Abstract: Cholesterol synthesis is essential for homeostasis of the epidermis, being required for both cell division and differentiation, as well as maintenance of the epidermal permeability barrier. Cholesterol synthesis in keratinocytes has been demonstrated to be regulated by sterol levels and the barrier function of the stratum corneum. Cholesterol synthesis in the epidermis is correlated with changes in mRNA levels for key enzymes, such as HMG‐CoA synthase and HMG‐CoA reductase, which have been previously demonstrated to be coordinately regulated by the sterol regulatory element binding proteins (SREBPs). In this study we demonstrate that a functional sterol regulatory element is required for sterol regulation of HMG‐CoA synthase in keratinocytes. We also investigate the regulation of cholesterol synthesis by fatty acids, which are another important constituent of the stratum corneum lipids. Palmitic and oleic acid inhibit 14C‐labelled acetate incorporation into sterols in a similar manner to sterols. However, unlike sterols, 50 μM oleic acid increase the steady state mRNA levels of HMG‐CoA synthase and the activity of the HMG‐CoA synthase promoter. The addition of 50 μM oleic acid to 25‐hydroxycholesterol results in an enhancement of the inhibitory effect of the sterol on promoter activity. The inhibition of acetate incorporation into sterols in human keratinocytes by 50 μM palmitic and 50 μM oleic acid is not due to regulation of HMG‐CoA synthase at the level of transcription.