bluuebunny/crossref_metadata_2025 huggingface dataset:
Hepatic steatosis in chronic hepatitis C: impact on response to anti‐viral treatment with peg‐interferon and ribavirin
Authors: GUIDI M. , MURATORI P. , GRANITO A. , MURATORI L. , PAPPAS G. , LENZI M. , BIANCHI F. B.
Published: October 2005

Background: There is increasing evidence that hepatic steatosis contributes to the progression of liver fibrosis, whereas its impact on the efficacy of anti‐viral treatment is still under investigation.

Aim: To evaluate the effect of steatosis on the outcome of combined anti‐viral treatment.

Methods: We studied 102 consecutive naive patients with chronic hepatitis C receiving combined anti‐viral therapy (peg‐interferonα‐2b and ribavirin).

Results: Fifty (49%) of 102 patients had evidence of hepatic steatosis (29 grade 1, 16 grade 2 and 5 grade 3). Sustained virological response was similar in patients with and without steatosis (58% vs. 56%); moreover, the grade of steatosis did not affect the rate of sustained virological response (grade 1: 58%, grade 2: 56% and grade 3: 60%). Patients with steatosis had significantly higher serum levels of aspartate transaminase, alanine transaminase andγ‐glutamyltransferase (P = 0.007, 0.004 and 0.03, respectively), higher histological activity (P = 0.03), more advanced stage of fibrosis (P = 0.0394) and more often hepatitis C virus genotype 3 (P = 0.04).

Conclusions: Our findings suggest that hepatic steatosis in chronic hepatitis C, irrespective of its grade, is not a negative prognostic factor of response to combined anti‐viral therapy, even when the histological and biochemical profile of the disease is more aggressive.
Review article: how and when to use ciclosporin in ulcerative colitis
Authors: DURAI D. , HAWTHORNE A. B.
Published: October 2005

Although colectomy for ulcerative colitis is curative, long‐term quality of life is reduced.

Intravenous ciclosporin 4 mg/kg/day has significant toxicity. There is now evidence that low‐dose ciclosporin (2 mg/kg daily by intravenous infusion, or 5–6 mg/kg daily in a twice daily oral dosage) has an acceptable safety profile, even when used in combination with corticosteroids. Drug dosage should be adjusted to the levels of 150–250 ng/mL initially (random levels during intravenous infusion, or trough levels for oral use).

Ciclosporin should be considered not only in those who have failed 7 days of corticosteroids, but also in fulminant colitis at day 3, if not responding to corticosteroids. The drug should be avoided in frail or elderly patients with significant comorbidity, and also where colectomy is likely to be necessary in the short to medium term. Ciclosporin should not be continued for more than 7 days, unless there is a definite response.

A 70–80% initial response is likely, and responders are discharged on oral ciclosporin, adding thiopurines and tailing prednisolone rapidly. The drug should be continued for 3 months. The likelihood of avoiding colectomy over 2–3 years is 40–50%.

More studies are needed to evaluate the use of oral ciclosporin in corticosteroid‐refractory colitis in out‐patients, and to assess whether monotherapy (without corticosteroids) is significantly safer, without loss of efficacy.
Molecular typing of adult acute myeloid leukaemia: significance of translocations, tandem duplications, methylation, and selective gene expression profiling
Authors: Olesen Lene Hyldahl , Aggerholm Anni , Andersen Bodil Lind , Nyvold Charlotte Guldborg , Guldberg Per , Nørgaard Jan Maxwell , Hokland Peter
Published: October 2005

Although a number of molecular aberrations have been described in acute myeloid leukaemia (AML), no study has yet determined their relative prognostic importance. We have analysed blast cells from 250 adult patients treated at the same institution during a 15‐year period. Balanced translocations were detected by multiplex polymerase chain reaction in 13% of the cases. Internal tandem duplication (ITD) of the FLT3 gene and partial tandem duplication of the MLL gene were found in 24% and 4%, respectively. Promoter hypermethylation of the P15, CDH1, ER and MDR1 genes was observed in 71%, 64%, 40% and 4%, respectively. Compared with normal bone marrow, the chemotherapy resistance protein MRP1 and apoptosis related genes BAX and CASPASE3 were found to be overexpressed in AML blasts. Univariate analysis revealed that the most important determinants of prognosis were presence of balanced translocations, age, white blood cell count and extramedullary disease, in order of statistical significance. In a multivariate analysis, balanced translocations retained their prognostic significance and FLT3 ITD as well as high gene expression of MDR1 were negative prognostic factors. From these data, which are the first to compare these molecular aberrations directly, we conclude that, when a battery of molecular changes is evaluated for upfront significance in AML, recurrent translocations are of prime importance for treatment outcome.
Childhood anaplastic large cell lymphoma has a high incidence of ALK gene rearrangement as determined by immunohistochemical staining and fluorescent in situ hybridisation: a genetic and pathological correlation*
Authors: Perkins Sherrie L. , Pickering Diane , Lowe Eric J. , Zwick David , Abromowitch Minnie , Davenport Ginny , Cairo Mitchell S. , Sanger Warren G.
Published: October 2005

Anaplastic large cell lymphoma (ALCL) comprises 10–15% of childhood non‐Hodgkin lymphomas (NHL). Systemic ALCL is highly associated with anaplastic lymphoma kinase (ALK) gene translocations with over‐expression of ALK protein. We studied ALK rearrangements using fluorescence in situ hybridisation (FISH) and ALK immunohistochemical staining in 43 paediatric systemic ALCLs. FISH (performed on 35 cases) identified a translocation in 29 cases (83%). Immunohistochemistry identified ALK over‐expression in 42/43 cases (97%) with the single ALK‐negative case demonstrating an ALK rearrangement by FISH, indicating 100% incidence of ALK translocations.
Lymphomatous polyposis of the gastrointestinal tract, including mantle cell lymphoma, follicular lymphoma and mucosa‐associated lymphoid tissue lymphoma
Authors: Kodama T , Ohshima K , Nomura K , Taniwaki M , Nakamura N , Nakamura S , Kohno S , Yamamoto J , Karube K , Yamasita Y , Shirakusa T , Kikuchi M
Published: October 2005

Aims:  Lymphomatous polyposis (LP) is considered to represent mantle cell lymphoma (MCL) of the gastrointestinal (GI) tract. However, a few reports have suggested that some are follicular lymphoma (FL) or mucosa‐associated lymphoid tissue (MALT) lymphomas. In this study, we analysed 35 patients and clarified the clinicopathological features of LP.

Methods and results:  Paraffin‐embedded tissue samples were stained immunohistochemically and analysed by tissue‐fluorescence in situ hybridization (T‐FISH) for IGH/CCND1 (cyclin D1) and IGH/BCL2. The average age of the patients was 58.3 years. Over half of the cases showed gastric, duodenal, small intestinal, ileocaecal and sigmoid colonic lesions (15, 19, 15, 16 and 16 cases, respectively). Phenotypically, cases were classified into three types of MCL (cyclin D1+ CD5+ CD10–) (n = 12), FL (cyclin D1– CD5– CD10+) (n = 14) and MALT (cyclin D1– CD5– CD10–) (n = 9). T‐FISH identified 11 of the 11 examined cases with MCLs to have IGH/CCND1, while seven of 10 cases with FL had IGH/BCL2, and none of the MALT cases were positive for IGH/CCND1 or IGH/BCL2. At the study endpoint, five of 12 patients with MCL were dead, two of 14 with FL and one of nine with MALT were dead of other disease. Event‐free survival analysis showed significantly poorest outcome in MCL, followed by FL, while MALT was associated with a favourable outcome (P = 0.0040).

Conclusions:  Our study emphasizes the importance of differentiating MCL, FL and MALT of LP in evaluating prognosis and hence the most suitable therapeutic regimen.
Histopathology of acute acalculous cholecystitis in critically ill patients
Authors: Laurila J J , Ala‐Kokko T I , Laurila P A , Saarnio J , Koivukangas V , Syrjälä H , Karttunen T J
Published: October 2005

Aims:  To illustrate the histopathological features of acute acalculous cholecystitis (AAC) of critically ill patients and to compare them with those of acute calculous cholecystitis (ACC) and normal gallbladders.

Methods and results:  We studied 34 gallbladders with AAC and compared them with 28 cases of ACC and 14 normal gallbladders. Histological features were systematically evaluated. Typical features in AAC were bile infiltration, leucocyte margination of blood vessels and lymphatic dilation. Bile infiltration in the gallbladder wall was more common and extended wider and deeper into the muscle layer in AAC compared with ACC. Epithelial degeneration and defects and widespread occurrence of inflammatory cells were typical features in ACC. Necrosis in the muscle layer was also more common and extended wider and deeper in ACC. There were no differences in the occurrence of capillary thromboses, lymphatic follicles or Rokitansky–Aschoff sinuses between the AAC and ACC samples.

Conclusions:  There are characteristic differences in histopathology between AAC and ACC, although due to overlap, none appeared to be specific as such for either condition. These results suggest that AAC is largely a manifestation of systemic critical illness, whereas ACC is a local disease of the gallbladder.
Primary invasive micropapillary carcinoma of the colon
Authors: Sakamoto K , Watanabe M , De La Cruz C , Honda H , Ise H , Mitsui K , Namiki K , Mikami Y , Moriya T , Sasano H
Published: October 2005

Aims:  Invasive micropapillary carcinoma (IMPC) is associated with frequent lymph node metastasis and adverse clinical outcome. IMPC has been reported in breast, urinary bladder, ureter, lung and parotid gland but not in colon. We present the clinicopathological features of three cases of primary IMPC of the colon with a review of the literature.

Methods and results:  The patients (one man and two women) were 53, 67 and 68 years old, respectively. The size of the tumour ranged from 20 to 100 mm in diameter. Histologically, all cases were composed predominantly of papillary tumour cell clusters with spaces in a background of fine fibrocollagenous stroma. One of the tumours (case 1) was nearly completely composed of IMPC, but the other two were associated with foci of adenocarcinoma and concurrent mucinous carcinoma, respectively. MUC1 was positive in all cases, suggestive of reverse cell orientation which is responsible for its unique histological features.

Conclusions:  We report three cases of primary IMPC of the colon. Its clinical significance remains undetermined but the presence of this component may represent a poor prognostic factor.
Breast pathology practice: most common problems in a consultation service
Authors: Putti T C , Pinder S E , Elston C W , Lee A H S , Ellis I O
Published: October 2005

Considerable progress has been made in understanding breast lesions utilizing molecular methods, but conventional morphology, simple immunohistochemical stains and common sense still prevail in diagnosing the vast majority of breast disease. The focus of this review is to identify the most common breast lesions sent to our consultation practice, and to reiterate salient diagnostic features, differential diagnoses and common pitfalls in identifying these lesions. Separation of epithelial proliferative lesions and differentiation between usual epithelial hyperplasia (UEH) and atypical ductal hyperplasia (ADH) are the most common problems encountered in our Consultation practice. Differentiation between UEH and ADH is based on the assumption that ADH is a clonal process, recognized by a uniform phenotype and more recently described immunohistochemical markers such as differential cytokeratin and also hormone receptor expression. Difficulty in subtyping invasive carcinomas and exclusion of in situ and/or invasive carcinoma in a sclerosing lesion is also commonly noted. Finally, problems in distinguishing various papillary and fibroepithelial lesions are also encountered. The use of common immunohistochemical stains such as various cytokeratin and myoepithelial markers, E‐cadherin and hormone receptors is helpful in solving most of these diagnostic dilemmas.
Expression of acyl‐CoA synthetase 5 in human endometrium and in endometrioid adenocarcinomas
Authors: Gassler N , Yang S H Y , Keith M , Helmke B M , Schirmacher P , Obermüller N
Published: October 2005

Aim:  Fatty acid metabolism of the endometrium is important for tissue homeostasis in the proliferative and secretory phase of the menstrual cycle. The enzyme acyl‐CoA synthetase 5 (ACS5) plays a crucial role in fatty acid metabolism, mainly through the generation of multifunctional long‐chain‐fatty‐acid‐CoA esters. The aim of the present study was to characterize expression and localization of ACS5 in the normal human endometrium and in endometrioid adenocarcinomas.

Methods and results:  Expression of ACS5 in the human endometrium was investigated by in situ techniques (immunohistochemistry, mRNA in situ hybridization) and a molecular approach (reverse transcriptase‐polymerase chain reaction, Western blot). ACS5 protein and mRNA were localized to the epithelium of the human endometrium. Here, ACS5 expression was found throughout the menstrual cylce as well as in the postmenopausal endometrium. Notably, in endometrioid adenocarcinomas, the ACS5 molecule was found abundantly in well‐differentiated tumours, but not in poorly differentiated adenocarcinomas.

Conclusions:  The abundance of ACS5 in the endometrial epithelium throughout the menstrual cycle provides support for its role in the regulation of tissue homeostasis. With regard to its value for histopathological diagnosis, immunohistochemical characterization of endometrioid adenocarcinomas shows that a decrease in ACS5 expression correlates with tumour dedifferentiation.
Histopathological features of chronic granulomatous disease (CGD) in childhood
Authors: Levine S , Smith V V , Malone M , Sebire N J
Published: October 2005

Aims : To describe the spectrum of histopathological features encountered in children with chronic granulomatous disease (CGD) at a specialist centre.

Methods and results : The histopathological findings of 88 surgical pathology requests from a range of organ systems including upper and lower gastrointestinal tract biopsy series, liver, bladder, bone, lung, skin, soft tissue, bone marrow and lymph node biopsy specimens, in 32 patients aged 4 months to 18 years (median 7 years) with CGD were reviewed. In most tissues the features were those of active chronic inflammation, with or without abscess or granuloma formation, often associated with fungal infection. In some tissues, more characteristic findings were identified, including the presence of pigmented macrophages, especially in hepatic sinusoids and colonic mucosa, where active chronic eosinophil predominant colitis was also observed.

Conclusions : Chronic granulomatous disease may present to histopathologists in a wide range of tissue specimens most often demonstrating features of active chronic inflammation with or without granuloma formation. The presence of numerous pigmented macrophages in association with such inflammation should raise suspicion of the diagnosis. In addition, diffuse granulomatous inflammation of the lung and hepatic abscess formation should be regarded as suggestive of the diagnosis.
CD117+ small cell lung cancer lacks the asp 816→val point mutation in exon 17
Authors: Mojica W D , Saxena R , Starostik P , Cheney R T
Published: October 2005

Aims : To determine the frequency of point mutation in c‐kit in CD117+ small cell lung cancer (SCLC). A significant proportion of SCLCs have been documented to be CD117+, thereby signifying they express the c‐kit gene product. This finding suggests this tumour may be a potential target for tyrosine kinase inhibitor (TKI) agents directed at c‐kit. A point mutation in exon 17 of the c‐kit gene, however, can abrogate the binding of TKIs. This being the case, immunohistochemistry is necessary to identify potential candidates for treatment with TKIs, but DNA sequence analysis may need to be performed to determine if these tumours will respond.

Methods and results : Tumour cells of 23 cases of SCLC showing immunoreactivity for CD117 were laser capture microdissected from archived formalin‐fixed paraffin‐embedded tissue and the DNA isolated. PCR on exon 17 of the c‐kit gene was performed and the amplified product sequenced. No point mutations were detected.

Conclusions : The absence of mutations in exon 17 of CD117+ SCLC suggests this tumour may respond to therapy with TKI.
Primary lymphoma arising in the nasal cavity among Japanese
Authors: Kitamura A , Yamashita Y , Hasegawa Y , Kojima H , Nagasawa T , Mori N
Published: October 2005

Aims : Most lymphomas arising in the nasal cavity are thought to be of natural killer (NK) cell origin. However, some reports indicate that T‐ and B‐cell lymphomas may also primarily arise in the nasal cavity. We therefore studied lymphomas arising in the nasal cavity both histologically and immunohistochemically.

Methods and results : Of the 32 cases investigated, 20 cases were also available as fresh frozen specimens. We diagnosed 31 cases as extranodal NK/T‐cell lymphoma and one as plasmacytoma. The neoplastic cells were immunoreactive for CD3 (polyclonal) 31/31, LMP‐1 12/31, CD20 (L26) 0/31, granzyme B 30/31, TIA‐1 30/30, CD56 (123C3) 29/31, CD4 0/31 and CD8 3/31. In situ hybridization for Epstein–Barr virus‐encoded small RNA‐1 (EBER‐1) was detected in 31/31. In frozen tissue sections, neoplastic cells mostly showed CD3 (Leu4)–, CD4 (Leu3a)–, CD5 (Leu1)–, CD8 (Leu2)–, CD16 (Leu11)–, CD56 (Leu19)+, βF1–, TCRδ1–, perforin+, CD94+ phenotypes. These immunohistochemical findings indicate their NK cell origin. In three cases, neoplastic cells were positive for CD8. In one of these cases, neoplastic cells were positive for CD8β and Vα24, suggesting their NKT‐cell origin.

Conclusions : Our present study indicates that primary lymphomas arising in the nasal cavity are mostly of NK cell derivation. Our present study also suggests that a small number of cases are derived from NKT‐cells.
Resistin gene 3′‐untranslated region +62G→A polymorphism is associated with hypertension but not diabetes mellitus type 2 in a German population
Authors: GOUNI‐BERTHOLD I. , GIANNAKIDOU E. , FAUST M. , KRATZSCH J. , BERTHOLD H. K. , KRONE W.
Published: October 2005

Objectives.  Resistin, a peptide hormone produced by adipocytes, has been associated with diabetes mellitus type 2 (DM‐2) in some rodent models. In humans the exact function of resistin remains unknown. Some, but not all studies have found associations between polymorphisms in the resistin gene with DM‐2. Recently a 3′‐untranslated region +62G→A polymorphism of the resistin gene has been associated with decreased risk for DM‐2 and for hypertension in diabetics in a Chinese population. Purpose of the present study was to examine for the first time in a German Caucasian population the possible association between this polymorphism and DM‐2, hypertension, lipoprotein levels, resistin levels as well as atherosclerosis.

Design, setting and subjects. A total of 818 subjects participated in the study. The presence of the +62G→A polymorphism of the resistin gene was investigated using polymerase chain reaction‐restriction fragment length polymorphism in 384 subjects with DM‐2 [224 men, 160 women, age 63.4 ± 10.6 years, body mass index (BMI) 28.7 ± 5.1 kg m−2] and in 434 nondiabetic age‐ and sex‐matched control subjects (248 men, 186 women, age 64.4 ± 6.5 years, BMI 26.5 ± 3.7 kg m−2).

Results.  Thirty‐four subjects were found to be carrying the +62G→A polymorphism in the control and 24 in the diabetic group (allelic frequencies 4% and 3.2% respectively). Subjects with DM‐2 were not found to have a different frequency of the genotypes (93.75% and 6.258%, for GG:GA/AA respectively) than the control subjects (92.2% and 7.8% for GG:GA/AA respectively) (OR 0.75, 95% CI 0.44–1.3, P = 0.31). In the total cohort, carriers of the A allele had a higher prevalence of hypertension (OR 1.82, 95% CI 1.03–3.21, P = 0.039). When analysed separately, the control group showed a strong association between the presence of the A allele and hypertension (OR 2.92, 95% CI 1.38–6.15, P = 0.005), whilst no such association could be established in the diabetic group (OR 1.05, 95% CI 0.43–2.54, P = 0.92). Multiple regression analysis confirmed that the presence of the A variant is associated with hypertension in control but not in diabetic subjects, independent of age and BMI. The polymorphism had no significant influence on the presence of atherosclerotic disease, BMI, and on triglyceride, HDL and LDL cholesterol levels, both, in the control and the diabetic groups. There was no difference in the serum resistin levels between the 62G→A variant carriers and noncarriers.

Conclusions.  In conclusion, the present data suggest that in a German Caucasian population the +62G→A polymorphism of the resistin gene is associated with hypertension but not with DM‐2.
HHV‐6‐related acute liver failure in two immunocompetent adults: favourable outcome after liver transplantation and/or ganciclovir therapy
Authors: CACHEUX W. , CARBONELL N. , ROSMORDUC O. , WENDUM D. , PAYE F. , POUPON R. , CHAZOUILLÈRES O. , GOZLAN J.
Published: October 2005

Fulminant hepatitis of unknown origin remain a significant cause of mortality, for which liver transplantation is often considered as the only therapeutic option. In retrospective studies, human herpesvirus 6 (HHV‐6) infections have been associated with such diseases, but the diagnosis of HHV‐6 infection of the liver is rarely established during the acute phase of liver failure. Using real‐time polymerase chain reaction (PCR), we diagnosed two cases of severe acute liver failure (ALF) related to HHV‐6 occurring in immunocompetent young adults. Both cases had a favourable outcome, one after valganciclovir therapy, one after liver transplantation associated with ganciclovir. Viral origin was evidenced in each case by the detection of high amounts of HHV‐6 DNA in liver tissue by the PCR assay. The decrease of intrahepatic viral load after therapeutic intervention was also monitored by quantitative PCR and paralleled in the two cases the clinical improvement. Diagnosis of HHV‐6 infection must be systematically evoked in case of unexplained ALF, since it might lead to specific therapeutic interventions, in addition of liver transplantation.
Bone mass in young adults: relationship with gender, weight and genetic factors
Authors: VALERO C. , ZARRABEITIA M. T. , HERNÁNDEZ J. L. , ZARRABEITIA A. , GONZÁLEZ‐MACÍAS J. , RIANCHO J. A.
Published: October 2005

Objectives.  To determine the relationship of the bone mass attained in young adults with anthropometric and genetic factors.

Design.  Cross‐sectional study of normal individuals.

Methods.  We studied 341 healthy subjects between 22 and 45 years of age. Bone mineral density (BMD) was measured by dual‐energy X‐ray absorptiometry (DXA) and correlated with body weight, height and nine polymorphisms in six genes involved in sex steroid metabolism (17‐hydroxylase, aromatase and 5‐reductase) and activity (oestrogen receptors (ER)‐α and ‐β, and androgen receptor).

Results.  The BMD was higher in men than in women (spine: 1.048 ± 0.120 vs. 1.034 ± 0.112; hip: 0.907 ± 0.131 vs. 0.822 ± 0.104 g cm−2, P < 0.001). However, the difference was due, at least in part, to the larger body size in men and diminished markedly after height adjustment. There was a negative correlation between age and hip BMD. Body weight was the single most influential factor on spine and hip BMD in both sexes, explaining 8–9% of BMD variance. Amongst the genetic factors studied, a common CA repeat polymorphism in ER‐β showed a significant association with BMD in women (P = 0.03 at the spine, and 0.008 at the hip). The relationship between ER‐β genotype and BMD persisted after adjustment by body weight and age, explaining a further 2–3% of BMD variance. Allelic variants of other genes studied were not related with BMD.

Conclusions.  Body weight and allelic variants of ER‐β are associated with BMD in young adults.
The biosynthesis and regulation of biosynthesis of Concord grape fruit esters, including ‘foxy’ methylanthranilate
Authors: Wang Jihong , Luca Vincenzo De
Published: October 2005

The biosynthesis of methyl anthranilate, the volatile compound responsible for the distinctive ‘foxy’ aroma and flavor of the Washington Concord grape (Vitis labrusca), involves an alcohol acyltransferase that catalyzes the formation of methyl anthranilate from anthraniloyl‐coenzyme A (CoA) and methanol. Although methanol is a poor substrate in comparison with the co‐substrate, high levels of this acyltransferase (0.5% of the total protein) combined with relatively high levels of this alcohol make this reaction possible in grapes. This 449 amino acid protein belongs to the BAHD family of acyltransferases, having 58% identity with the benzoyl CoA:benzyl alcohol benzoyl transferase from Clarkia. Both native and recombinant enzymes can use a broad range of acyl‐CoAs and alcohols as substrates. The ability of Concord grape alcohol acyltransferase to accept a range of different CoA esters and alcohols suggests this to be a versatile ester‐forming enzyme, similar to those of other fruits that than can produce a range of fruit esters based on the supply of appropriate substrates. Expression is coordinately regulated, with transcript, protein and enzyme activities coinciding with the accumulation of methyl anthranilate that occurs after the initiation of berry ripening. The majority of acyltransferase protein in grape tissues is localized to the outer fruit mesocarp, a result consistent with the fact that methyl anthranilate is released to the external environment throughout the ripening process. Wine grapes (Vitis vinifera) that accumulate neither anthranilate nor methyl anthranilate do not express this enzyme activity nor do they accumulate this protein.
Localization of tabersonine 16‐hydroxylase and 16‐OH tabersonine‐16‐O‐methyltransferase to leaf epidermal cells defines them as a major site of precursor biosynthesis in the vindoline pathway in Catharanthus roseus
Authors: Murata Jun , Luca Vincenzo De
Published: October 2005

The Madagascar periwinkle (Catharanthus roseus) produces the well known and remarkably complex anticancer dimeric alkaloids vinblastine and vincristine, which are derived by the coupling of vindoline and catharanthine monomers. Recent data from in situ RNA hybridization and immunolocalization suggest that combinatorial cell factories within the leaf are involved in vindoline biosynthesis. In this study, the cell types responsible for vindoline biosynthesis were identified by laser‐capture microdissection/RNA isolation/RT–PCR to show that geraniol hydroxylase, secologanin synthase, tryptophan decarboxylase, strictosidine synthase, strictosidine ß‐glucosidase and tabersonine 16‐hydroxylase can be detected preferentially in epidermal cells. A new and complementary application of the carborundum abrasion (CA) technique was developed to obtain epidermis‐enriched leaf extracts that can be used to measure alkaloid metabolite levels, enzyme activities and gene expression. The CA technique showed that tabersonine and 16‐methoxytabersonine, together with 16‐hydroxytabersonine‐16‐O‐methyltransferase, are found predominantly in Catharanthus leaf epidermis, in contrast to vindoline, catharanthine and later enzymatic steps in vindoline biosynthesis. The results show that leaf epidermal cells are biosynthetically competent to produce tryptamine and secologanin precursors that are converted via many enzymatic transformations to make 16‐methoxytabersonine. This alkaloid or its 2,3 dihydro‐derivative is then transported to cells (mesophyll/idioblast/laticifer) within Catharanthus leaves to complete the last three or four enzymatic transformations to make vindoline.
The immediate‐early ethylene response gene OsARD1 encodes an acireductone dioxygenase involved in recycling of the ethylene precursor S‐adenosylmethionine
Authors: Sauter Margret , Lorbiecke René , OuYang Bo , Pochapsky Thomas C. , Rzewuski Guillaume
Published: October 2005

Methylthioadenosine (MTA) is formed as a by‐product of ethylene biosynthesis from S‐adenosyl‐l‐methionine (AdoMet). The methionine cycle regenerates AdoMet from MTA. In two independent differential screens for submergence‐induced genes and for 1‐aminocyclopropane‐1‐carboxylic acid (ACC)‐induced genes from deepwater rice (Oryza sativa L.) we identified an acireductone dioxygenase (ARD). OsARD1 is a metal‐binding protein that belongs to the cupin superfamily. Acireductone dioxygenases are unique proteins that can acquire two different activities depending on the metal ion bound. Ectopically expressed apo‐OsARD1 preferentially binds Fe2+ and reconstituted Fe‐OsARD1 catalyzed the formation of 2‐keto‐pentanoate and formate from the model substrate 1,2‐dihydroxy‐3‐ketopent‐1‐ene and dioxygen, indicating that OsARD1 is capable of catalyzing the penultimate step in the methionine cycle. Two highly homologous ARD genes were identified in rice. OsARD1 mRNA levels showed a rapid, early and transient increase upon submergence and after treatment with ethylene‐releasing compounds. The second gene from rice, OsARD2, is constitutively expressed. Accumulation of OsARD1 transcript was observed in the same internodal tissues, i.e. the meristem and elongation zone, which were previously shown to synthesize ethylene. OsARD1 transcripts accumulated in the presence of cycloheximide, an inhibitor of protein synthesis, indicating that OsARD1 is a primary ethylene response gene. Promoter analysis suggests that immediate‐early regulation of OsARD1 by ethylene may involve an EIN3‐like transcription factor. OsARD1 is induced by low levels of ethylene. We propose that early feedback activation of the methionine cycle by low levels of ethylene ensures the high and continuous rates of ethylene synthesis required for long‐term ethylene‐mediated submergence adaptation without depleting the tissue of AdoMet.
Efficient insertional mutagenesis in rice using the maize En/Spm elements
Authors: Kumar Chellian Santhosh , Wing Rod A. , Sundaresan Venkatesan
Published: October 2005

We have developed a novel system for insertional mutagenesis in rice (Oryza sativa) based on the maize (Zea mays) enhancer/suppressor mutator (En/Spm) element. In this system, a single T‐DNA construct with Spm‐transposase and the non‐autonomous defective suppressor mutator (dSpm) element is used in conjunction with green fluorescent protein (GFP) and Discosoma sp. Red Fluorescence Protein (DsRed) fluorescent markers to select unlinked stable transpositions of dSpm. Using this system, we could demonstrate high frequencies of unlinked germinal transposition of dSpm in rice. Analysis of dSpm flanking sequences from 353 stable insertion lines revealed that the dSpm insertions appear to be widely distributed on rice chromosomes with a preference for genic regions (70%). The dSpm insertions appear to differ from Activator‐Dissociation (Ac‐Ds) elements in genomic distribution and exhibit a greater fraction of unlinked transpositions when compared with Ds elements. The results obtained in this study demonstrate that the maize En/Spm element can be used as an effective tool for functional genomics in rice and can complement efforts using other insertional mutagens. Further, the efficacy of the non‐invasive fluorescence‐based selection system is promising for its application to other crops.
The virtual crossmatch – A screening tool for sensitized pediatric heart transplant recipients
Authors: Zangwill S. D. , Ellis T. M. , Zlotocha J. , Jaquiss R. D. , Tweddell J. S. , Mussatto K. A. , Berger S.
Published: October 2005

Abstract:  Heart transplantation in the setting of human leukocyte antigen (HLA) sensitization is challenging, as a time‐consuming prospective crossmatch (XM) may be required, severely limiting the number of potential donors. We evaluated a ‘virtual XM’, defining a positive virtual XM as the presence of recipient pre‐formed anti‐HLA antibodies to the prospective donor HLA type, and compared the virtual XM to a standard direct XM. Bead‐based flow cytometric analysis was used to identify anti‐HLA antibody (Ab) present in a child listed for heart transplantation. Using recipient serum, direct‐flow cytometric T‐ and B‐cell XM were run for potential donors against whose HLA type the recipient had specific antibodies (group 1, n = 7) and for potential donors with predicted compatible HLA types by virtual XM (group 2, n = 7). Results were expressed as median channel difference (MCD) between the control and recipient serum. A positive T‐cell XM was defined as MCD > 50, whereas MCD > 100 constituted a positive B‐cell result. The rate of T‐cell reactivity was significantly less in group 2 than in group 1 (29% vs. 100%, p = 0.02); similarly, B‐cell reactivity was also less for group 2 (14% vs. 100%, p = 0.005). The virtual XM was 100% sensitive in detecting positive flow cytometric XM results for T and B cells. Although only 72% specific in predicting a negative T‐cell XM, and 86% specific for negative B‐cell XM, the false negatives were weakly positive and would probably have been clinically acceptable. Currently, potentially suitable donor organs are often declined for lack of a prospective XM; these organs may ultimately be allocated to more distant recipients or perhaps not used at all. While further studies are needed, virtual XM has the potential to improve availability of organs for sensitized patients and improve the overall allocation process.
Hepatocellular carcinoma in children and effect of living‐donor liver transplantation on outcome
Authors: Arikan C. , Kilic M. , Nart D. , Ozgenc F. , Ozkan T. , Tokat Y. , Yagci R. V. , Aydogdu S.
Published: October 2005

Abstract:  Hepatocellular carcinoma (HCC) is primarily observed in the older children and in most cases it develops in association with liver cirrhosis. Liver transplantation offers a good chance for long‐term cure. To evaluate the outcome of children with HCC and the impact of living‐donor orthotopic liver transplantation (OLT) on survival a retrospective review of radiographic, laboratory, pathologic, and therapeutic data in 13 children (six female and seven male) with chronic liver disease accompanied with HCC were studied. The patients were divided into two groups according to therapeutic modality: transplanted and non‐transplanted patients. Kaplan–Meier survival curves in various therapeutic groups were plotted. The mean age of patients was 6.4 ± 4.8 yr. Pediatric end‐stage liver disease score was adapted to model for end‐stage liver disease score for HCC and ranged between 1–44 and 18–44, respectively. The underlying liver diseases were tyrosinemia type 1 (n = 6), chronic hepatitis B infection (n equals;6), glycogen storage disease type 1 (n = 1). Alfa‐feto protein levels were elevated in all patients except one. Median number of tumor nodules was three (1–10), median maximal diameter of tumor nodules was 3.4 cm (0.5–8). Eleven patients were eligible for OLT whereas two patients were not eligible. Seven of the 11 patients considered for transplantation underwent living‐donor OLT. Remaining four patients died while waiting on cadaveric transplant list. Overall 1 and 4‐yr survival rates for all patients were 53.3 and 26.6%, respectively, and were found significantly higher in transplanted children than non‐transplanted children (72%, 72% vs. 33% and 16.6%). No patient had tumor recurrence at median of 36‐month follow‐up after OLT. OLT is a life‐saving procedure for children with chronic liver disease accompanying with HCC. Living‐donor OLT avoids the risk of tumor progression and transplant ineligibility in these children.
Infections in children undergoing allogeneic bone marrow transplantation in India
Authors: George Biju , Mathews Vikram , Viswabandya Auro , Srivastava Alok , Chandy Mammen
Published: October 2005

Abstract:  We studied the clinical profile of infections among 221 pediatric patients who underwent 230 allogeneic transplants between 1986 and June 2004. All patients developed febrile neutropenia. There were 283 documented infections, which included bacterial (36.9%), viral (45.7%), fungal (11.1%) and other infections (6.3%) including tuberculosis. Bacterial and fungal infections were more common in the first 30 days following BMT, while viral infections were more common >30 days after BMT. Bacterial pathogens were predominantly gram‐negative organisms (72.7%), when compared with gram‐positive organisms (27.3%). Common gram‐negative organisms included NFGNB, Pseudomonas, Escherichia coli and Klebsiella while coagulase negative Staphylococci was the main gram‐positive organism. Bacteremia (61.2%) was the main source positive cultures and was mainly because of gram‐negative organisms (81%), predominantly NFGNB and Pseudomonas. Exactly 103/221(43.7%) transplants had 128 documented viral infections commonly because of Cytomegalovirus, Herpes group of viruses and transfusion related hepatitis. Thirty of 221 (13.5%) of transplants had 30 documented fungal infections with the majority being because of aspergillus (90%). Tuberculosis was seen in 1.7% of transplants while catheter infections were seen in 21 patients (9.1%). Infection related mortality was seen in 12% predominantly because of CMV or fungal infections. A sub group analysis (pre‐1998 vs. post‐1998) revealed higher incidences of gram‐negative infections, bacteremia and bacterial infection related mortality in the pre‐1998 era when compared with the recent times. The profile and mortality of infections in this series from India is not significantly different from reports from the West.
Live virus immunization after orthotopic liver transplantation
Authors: Khan Saba , Erlichman Jessi , Rand Elizabeth B.
Published: October 2005

Abstract:  Viral infection is a recognized and potentially serious complication in children following solid organ transplantation. The risks of viral infection are particularly important for infants who may not have completed standard childhood immunizations at the time of transplantation and are therefore at risk for otherwise preventable infections. The general practice of withholding live virus immunization from transplant recipients has been questioned and several small studies have looked at MMR and/or Varivax administration in children following transplantation. This retrospective study analyzes the response to primary MMR and varicella immunization in selected pediatric liver transplant recipients in the largest such study to date. Nineteen of 26 children (73%) developed serologic immunity for measles following MMR (although 18 required multiple doses). Similarly, varicella immunization resulted in seroconversion in 20 of 31 children (64.5%; seven required multiple doses). Only minor adverse effects reported in the general population were observed. Live virus immunization with MMR and Varivax was safe and immunogenic in this selected population of liver transplant recipients.
Epstein–Barr viremia levels after pediatric liver transplantation as measured by real‐time polymerase chain reaction
Authors: Kullberg‐Lindh Carola , Ascher Henry , Saalman Robert , Olausson Michael , Lindh Magnus
Published: October 2005

Abstract:  Effective immunosuppression has improved the results following liver transplantation, but also increased the risk for opportunistic infections. Epstein–Barr virus (EBV) infection in transplant patients can cause various symptoms including the life‐threatening premalignant condition, post‐transplantation lymphoproliferative disorder (PTLD). Serum specimens from 24 consecutive children (mean 7.6 specimens/patient), who had undergone liver transplantation in Göteborg from January 1995 to May 2002, were analyzed retrospectively for EBV DNA by real‐time TaqMan® polymerase chain reaction (PCR). The results were related to clinical picture, immunosuppression, graft rejection and infections with other agents. Eleven patients (46%) developed primary EBV infection at a mean time of 4.8 months after transplantation, and six (25%) reactivated EBV infection at a mean of 4.0 months after transplantation. Four of the 11 patients with primary infection had symptomatic EBV infection: two had PTLD and two hepatitis. One patient in the group with reactivated infection developed PTLD. EBV DNA levels were significantly higher in the group with primary symptomatic infection compared with the patients with primary asymptomatic infection (mean 65 500 copies/mL; range 14 200–194 300 vs. 3700 copies/mL; range 100–9780). In patients with symptomatic infection EBV DNA levels did not differ between PTLD and hepatitis patients. The data suggest that quantitative analysis of EBV DNA in serum by real‐time PCR is useful for identification of EBV‐related disease.
Risk factors for reduced areal bone mineral density in young adults with stem cell transplantation in childhood
Authors: Taskinen Mervi , Kananen Kristiina , Välimäki Matti , Löyttyniemi Eliisa , Hovi Liisa , Saarinen‐Pihkala Ulla , Lipsanen‐Nyman Marita
Published: October 2005

Abstract:  Slightly, but significantly, reduced bone mineral density (BMD) has been detected as a late effect after stem cell transplantation (SCT) performed in childhood. The aim of the study was to evaluate the risk factors of reduced BMD after SCT in childhood. We evaluated areal BMD of 16 young adults (six males, 10 females), aged 21 yr (range 15–34) by dual‐energy X‐ray absorptiometry at the lumbar spine, at the femoral neck, in the total hip, and in the total body. Bone turnover rate was evaluated by markers of bone formation and resorption. Six of the 16 patients had reduced BMD with a Z‐score of ≤−1 at least at one measurement site. Factors associated with reduced BMD were prepubertal status at transplant (p = 0.03), delayed pubertal growth (p = 0.03), pubertal onset gonadal hormone insufficiency (p = 0.02), and female sex (p = 0.02). Surprisingly, height in SDs and lumbar spine BMD correlated negatively (p = 0.008) in those with reduced bone mass, indicating that low areal density could not be due the small size of the vertebrae. Bone turnover markers were similar for those with normal and reduced BMD. In conclusion, 38% of the SCT long‐term survivors had reduced areal BMD. Prepubertal status at transplant with pubertal onset gonadal hormone insufficiency and female sex predisposed to reduced bone mass after SCT performed in childhood.
Small bowel transplantation: Literature review 2003–2005
Authors: Botha Jean F. , Horslen Simon P.
Published: October 2005

Abstract:  The rapid expansion of the volume of peer reviewed publications in recent years, including a large increase in the number of new biomedical journals, makes it difficult for the practicing clinician to stay up to date with the medical literature. This review is part of a new series that will provide succinct summaries of the recent medical literature in the field of pediatric transplantation. In this review, we summarize important articles in the field of pediatric intestine transplantation that have been published over the last 2½ years (2003–2005). The review is intended to be comprehensive but not exhaustive.
Disseminated adenovirus infection with respiratory failure in pediatric liver transplant recipients: Impact of intravenous cidofovir and inhaled nitric oxide
Authors: Wallot M. A. , Dohna‐Schwake C. , Auth M. , Nadalin S. , Fiedler M. , Malagó M. , Broelsch C. , Voit T.
Published: October 2005

Abstract:  Adenoviruses (AdV) are opportunistic pathogens that can lead to severe infections and respiratory failure (acute respiratory distress syndrome, ARDS) with high mortality in immunosuppressed patients. Cidofovir (CDV) has been used in adenoviral disease in bone marrow transplant recipients. Two pediatric liver transplant recipients with disseminated adenoviral disease and ARDS were treated with reduction of immunosuppression, CDV, and inhaled nitric oxide (iNO). CDV 1 mg/kg was given three times per week intravenously with intravenous hydration and oral probenecid. Viral suppression and clinical improvement was achieved. AdV hepatitis did not occur, and graft function was preserved, although acute rejection occurred in both patients. Adverse effects were mild and transient not requiring dose modification. Severe hypoxemia was reversed with iNO 10–20 p.p.m. CDV treatment of AdV infections in organ transplant recipients and other immunocompromised patients should be further investigated in prospective studies.
Idiopathic infantile arterial calcification: Two case reports, a review of the literature and a role for cardiac transplantation
Authors: Glatz Andrew C. , Pawel Bruce R. , Hsu Daphne T. , Weinberg Paul , Chrisant Maryanne R. K.
Published: October 2005

Abstract:  Idiopathic infantile arterial calcification (IIAC) is a rare, but important, cause of rapidly progressive ischemic heart disease in children. In this paper, we report two recent cases of IIAC seen at tertiary referral hospitals. Both cases presented in infancy with signs of heart failure and, ultimately, died with the diagnosis of IIAC confirmed at postmortem examination. A thorough review of the literature reveals approximately 160 reported cases of IIAC. The clinical outcomes, radiographic findings and pathologic details are summarized. Proposed etiologic mechanisms are reviewed, including promising research into the role of inorganic pyrophosphate as a regulatory factor in the development of IIAC. Because of the typically fatal outcome of IIAC and the lack of proven therapies, the potential role for cardiac transplantation is discussed.
Peripheral blood mononuclear cell responses to heat shock proteins in patients undergoing stem cell transplantation
Authors: Sedlackova Lucie , Sedlacek Petr , Hromadnikova Ilona
Published: October 2005

Abstract:  GVHD is a major complication after allogeneic SCT. Etiology of GVHD is multifactorial. Known role of hSPS in antigen presentation could suggest their potential role in the alloreactive process that leads to aGVHD. HSPS represent major immunodominant antigens in a wide spectrum of microbial pathogens. Bacterial and fungal colonization, infection and sepsis are frequent in immunocompromised patients with various malignant and non‐malignant diseases. We studied PBMC responses to recombinant human hsp60 (rh‐hsp60), rh‐hsp70 and Mycobacterium bovis hsp65 (M. bovis hsp65) in relation to aGVHD and infection in 34 pediatric patients with various lympho‐hemopoietic malignancies as well as non‐malignant disorders subjected to SCT. PBMC of patients before initiation of preparative regimen as well as after engraftment were stimulated with hSPS (1 μg/mL/well, 7‐day cultivation). PHA was used as a control of the stimulation ability. Cell responses were measured after the incorporation of 3H‐thymidin (pulsing with 1 μCi/well) and were expressed as stimulation indexes (SI). We demonstrated significantly high proliferative response to rh‐hsp60 as well as M. bovis hsp65 in a cohort of pretransplant patients with anamnestic and/or actual infection when compared with a cohort of patients without infection and healthy individuals. Strong PBMC cell responses to hSPS were found in patients who were at present colonized with Escherichia coli and Klebsiella pneumoniae or had previously K. pneumoniae infection with subsequent sepsis. Our findings support various studies dealing with immunodominant hSPS in connection with several pathogens and infectious diseases. Although no statistical difference for proliferative response to PHA was observed, PBMC responses against all tested hSPS comparing a cohort of patients with aGVHD and that with no sign of GVHD resulted in significantly lower SI for all tested hSPS in patients with aGVHD. Lower stimulation with hSPS during aGVHD might be explained by the stress‐induced upregulation of self‐hSPS synthesis that might lead to the inhibition of self‐hSPS reactive T‐cell response. Vice versa, we hypothesize that increased hsp‐specific stimulation may reflect the presence of protecting regulatory T cells preventing the development of Th1‐mediated diseases involving aGVHD.