bluuebunny/crossref_metadata_2025 huggingface dataset:
A study of the nature of embryonic lethality in LIS1−/− Mice
Authors: Cahana A. , Jin X.L. , Reiner O. , Wynshaw‐Boris A. , O'Neill C.
Published: August 2003

Homozygous deletion of the Lis1 gene (Lis1−/−) in mouse resulted in early embryonic lethality immediately after embryo implantation by an undefined mechanism. We seek to define the nature of this demise. LIS1 (pafah1b1) is a 46 kDa protein with seven tryptophan‐aspartate (WD) repeats. It docks with many proteins and has been implicated in microtubular function, cell division, intercellular transport, and nuclear and cellular motility. Combined Western and quantitative reverse transcriptase‐polymerase chain reaction (RT‐PCR) analyses showed that LIS1 expression from the blastocyst stage required new transcription from the embryonic genome. Consequently, the death of post‐implantation embryos may not reflect the first time during development that LIS1 was required, rather, it may reflect the first time following depletion of gametic stores that its actions were essential. Following culture of blastocysts in vitro for 96 hr the inner cell mass (ICM) of null embryos were significantly smaller than ICM of wild‐type siblings. Normal blastocyst outgrowths after 96‐hr culture had high levels of LIS1 expression in the outer cells of developing ICM and extensive expression in trophoblast cells. Lis1−/− embryos had significantly smaller trophoblast nuclei than wild‐type embryos. The results show that LIS1 expression is required for the continued normal development of the ICM and optimal trophoblast giant cell formation. Mol. Reprod. Dev. 66: 134–142, 2003. © 2003 Wiley‐Liss, Inc.
Evaluation of gene targeting by homologous recombination in ovine somatic cells
Authors: Williams Sarah H. , Sahota Virender , Palmai‐Pallag Timea , Tebbutt Scott J. , Walker Jean , Harris Ann
Published: August 2003

Mouse models for some human genetic diseases are limited in their applications since they do not accurately reproduce the phenotype of the human disease. It has been suggested that larger animals, for example sheep, might produce more useful models, as some aspects of sheep physiology and anatomy are more similar to those of humans. The development of methods to clone animals from somatic cells provides a potential novel route to generate such large animal models following gene targeting. Here, we assess targeting of the cystic fibrosis transmembrane conductance regulator (CFTR) gene in ovine somatic cells using homologous recombination (HR) of targeting constructs with extensive (>11 kb) homology. Electroporation of these constructs into ovine fetal and post‐natal fibroblasts generated G418‐resistant clones, but none analyzed had undergone HR, suggesting that at least for this locus, it is an extremely inefficient process. Karyotyping of targeted ovine fetal fibroblasts showed them to be less chromosomally stable than post‐natal fibroblasts, and, moreover, extended culture periods caused them to senesce, adversely affecting their viability for use as nuclear transfer donor cells. These data stress the importance of donor cell choice in somatic cell cloning and suggest that culture time be kept to a minimum prior to nuclear transfer in order to maximize cell viability. Mol. Reprod. Dev. 66: 115–125, 2003. © 2003 Wiley‐Liss, Inc.
Impact of oxidative stress in aged mouse oocytes on calcium oscillations at fertilization
Authors: Takahashi Toshifumi , Takahashi Eiji , Igarashi Hideki , Tezuka Naohiro , Kurachi Hirohisa
Published: August 2003

In vivo post‐ovulatory aging of oocytes significantly affects the development of oocytes and embryos. Also, oocyte aging alters the regulation of the intracellular calcium concentration, thus affecting Ca2+ oscillations in fertilized oocytes. Because reactive oxygen species (ROS) are known to significantly perturb Ca2+ homeostasis mainly through direct effects on the machinery involved in intracellular Ca2+ storage, we hypothesized that the poor development of aged oocytes that may have been exposed to oxidative stress for a prolonged time might arise from impaired Ca2+‐oscillation‐dependent signaling. The fertilization rates of aged oocytes and of fresh oocytes treated with 100 μM hydrogen peroxide (H2O2) for 10 min were significantly lower than that of fresh oocytes. Comparing within the fertilized oocytes, blastocyst formation was decreased while embryo fragmentation was increased similarly in the aged and H2O2‐treated fresh oocytes. The frequency of Ca2+ oscillations was significantly increased whereas the amplitude of individual Ca2+ transients was lowered in the aged and H2O2‐treated fresh oocytes. The rates of rise and decline in individual Ca2+ transients were decreased in these oocytes, indicating impaired Ca2+ handling. When lipid peroxidation was assessed using 4,4‐difluoro‐5‐(4‐phenyl‐1,3‐buttadienyl)‐4‐bora‐3a, 4a‐diaza‐s‐indacene‐3‐undecanoic acid (C11‐BODIPY) in unfertilized oocytes placed in a 5% CO2 in air atmosphere, the green fluorescence (indicating lipid peroxidation) increased faster in the aged oocytes than in the fresh oocytes. Furthermore, the green fluorescence in the aged oocytes was already approximately 20 times higher than that in the fresh oocytes at the beginning of the measurements. These findings support the idea that Ca2+ oscillations play a key role in the development of fertilized aged oocytes. Mol. Reprod. Dev. 66: 143–152, 2003. © 2003 Wiley‐Liss, Inc.
Differential expression of lysosomal associated membrane protein (LAMP‐1) during mammalian spermiogenesis
Authors: Moreno Ricardo D.
Published: August 2003

The mammalian acrosome is a secretory vesicle of mature sperms that plays an important role in fertilization. Recent evidence had pointed out that some components found at endosomes in somatic cells are associated with the developing acrosome during the early steps of spermiogenesis. Moreover, the mammalian acrosome contains many enzymes found within lysosomes in somatic cells. In this work, we studied the dynamics of some components of the endosome/lysosome system, as a way to understand the complex membrane trafficking circuit established during spermatogenesis. We show that the cation independent‐mannose‐6‐phosphate receptor (CI‐MPR) is transiently expressed in the cytoplasm of mid‐stage spermatids (steps 5–11). On the other hand, γ‐adaptin, an adaptor molecule of a complex involved in trafficking from the Golgi to lysosomes, was expressed in cytoplasmic vesicles only in pachytene and Cap‐phase spermatids (steps 1–5). Our major finding is that the lysosomal protein LAMP‐1 is differentially expressed during spermiogenesis. LAMP‐1 appears late in spermatogenesis (Acrosome‐phase) contrasting with LAMP‐2, which is present throughout the complete process. Both proteins appear to be associated with cytoplasmic vesicles and not with the developing acrosome. None of the studied proteins is present in epididymal spermatozoa. Our results suggest that the CI‐MPR could be involved in membrane trafficking and/or acrosomal shaping during spermiogenesis. Mol. Reprod. Dev. 66: 202–209, 2003. © 2003 Wiley‐Liss, Inc.
β‐Alanine but not taurine can function as an organic osmolyte in preimplantation mouse embryos cultured from fertilized eggs
Authors: Hammer Mary‐Anne , Baltz Jay M.
Published: August 2003

Early preimplantation mouse embryos are susceptible to the detrimental effects of increased osmolarity and, paradoxically, their in vitro development is significantly compromised by osmolarities near that of oviductal fluid. In vitro development can be restored, however, by several compounds that are accumulated by 1‐cell embryos to act as organic osmolytes, providing intracellular osmotic support and cell volume regulation. Taurine, a substrate of the β‐amino acid transporter that functions as an organic osmolyte transporter in other cells, had been proposed to function as an organic osmolyte in mouse embryos. Here, however, we found that taurine is neither able to provide protection for in vitro embryo development against increased osmolarity nor is it accumulated to higher intracellular levels as osmolarity is increased, indicating that it cannot function as an organic osmolyte in early preimplantation embryos. In contrast, β‐alanine, the other major substrate of the β‐amino acid transporter, both protects against increased osmolarity and is accumulated to somewhat higher levels as osmolarity is increased, indicating that it is able to function as an organic osmolyte in embryos. However, we also found that β‐alanine is displaced from embryos by glycine—the most effective organic osmolyte in embryos previously identified—and β‐alanine does not increase protection above that afforded by glycine at concentrations near those in vivo. Thus, the β‐amino acid transporter is likely present in early preimplantation embryos to supply β‐amino acids such as taurine for purposes other than to serve as organic osmolytes. Mol. Reprod. Dev. 66: 153–161, 2003. © 2003 Wiley‐Liss, Inc.
Evidence for multiple distinctly localized adenylyl cyclase isoforms in mammalian spermatozoa
Authors: Baxendale Rhona W. , Fraser Lynn R.
Published: August 2003

In addition to a bicarbonate‐regulated soluble adenylyl cyclase (sAC), mammalian spermatozoa, like somatic cells, appear to contain receptor/G protein‐regulated AC activity that contributes to the modulation of specialized cell processes. This study provides evidence that agents, known to influence somatic membrane‐associated AC (mAC) but apparently not germ cell sAC, can modulate cAMP production and functional state in mouse spermatozoa. Specifically, forskolin significantly enhanced cAMP production and capacitation, while inclusion of 2′,5′‐dideoxyadenosine significantly blocked these responses. Furthermore, GTPγS and NaF stimulated cAMP, but GDPβS and mastoparan had no apparent effect, consistent with recent evidence that Gs, but not Gi, contributes to AC/cAMP regulation in uncapacitated cells. In addition, intact mouse spermatozoa were screened for all known mAC isoforms by immunolocalization, using commercially available specific antibodies. The most abundant isoforms appeared to be AC2, AC3, and AC8, each with distinct distributions in the acrosomal and flagellar regions; AC1 and AC4 also appeared to be present, although less abundantly, in the midpiece and acrosomal cap regions, respectively. Intriguingly, however, Western blotting revealed that the major immunoreactive proteins in mouse sperm lysates were considerably smaller (∼50–60 kDa) than their somatic cell counterparts, suggesting that mature spermatozoa contain multiple mACs which may function in a shortened form. Of particular interest were AC3 and AC8, located in the same regions as, and hence possibly directly associated with, specific cell surface receptors and G proteins that are able to regulate the spermatozoon's acquisition and maintenance of fertilizing ability via changes in AC/cAMP. Mol. Reprod. Dev. 66: 181–189, 2003. © 2003 Wiley‐Liss, Inc.
Experimental demonstration that pre‐ and post‐conceptional mechanisms influence sex ratio in mouse embryos
Authors: JimÉnez Adela , FernÁndez RaÚl , Madrid‐Bury Ninoska , Moreira Pedro Nuno , Borque Concepción , Pintado BelÉn , GutiÉrrez‐AdÁn Alfonso
Published: August 2003

Previously we have demonstrated in two monotocous species (bovine and sheep), a relationship between time of insemination, moment of ovulation, and embryo sex ratio. Here, we have analyzed in a polytocous specie (mice) if in addition to pre‐conceptional mechanisms, also post‐conceptional ones affect the offspring sex ratio. To verify this hypothesis we carried out two experiments. In the first experiment, we analyzed the effect of mating dynamics on the sex ratio of mice with synchronic male and female embryo development. Females were mated before and after ovulation and sacrificed 13 days later for sex determination of embryos and reabsorptions. A decreased litter size, and an increased offspring sex ratio in matings occurring later in oestrus, supported the view that a biased sex ratio may occur as the result of behavioral differences between the populations of X‐ or Y‐bearing spermatozoa. In the second experiment, embryos developmentally synchronic and asynchronic with the recipient female endometrium were transferred, and again, 13 days later, females were sacrificed for sex determination of embryos and reabsorptions. The male proportion per litter found, indicated that our developmentally asynchronic transfers favored a sex ratio disbalance at birth. When combined, these results become the first experimental evidence supporting the view that both pre‐ and post‐conceptional mechanisms of sex ratio distortion in polytocous species are not mutually exclusive and both may explain, under different conditions, sex ratio deviations at birth. Mol. Reprod. Dev. 66: 162–165, 2003. © 2003 Wiley‐Liss, Inc.
Spermatogenesis of mice lacking CK2α′: Failure of germ cell survival and characteristic modifications of the spermatid nucleus
Authors: Escalier Denise , Silvius Derek , Xu Xin
Published: August 2003

Csnk2a2 encodes the CK2α′catalytic subunit of CK2 that is predominantly expressed in testis. Male mice in which Csnk2a2 has been disrupted were infertile and displayed oligozoospermia with an abnormal shape of the spermatid nucleus. In this study, Csnk2a2 null testes revealed extensive germ cell degenerative processes at all stages of spermatogenesis, including the first spermatogenesis wave. Nuclear envelope (NE) protrusions with loss of nuclear pores, swelling of the outer membrane, and disruption of the inner membrane were observed in cells ranging from spermatogonia to early spermatids. Most early round spermatids were depleted, and DNA‐specific fluorescent dyes showed a large chromatin‐free nuclear domain near the chromocenter. Spermatids that were not eliminated retained NE defects that could explain the acrosomal and nuclear abnormalities of Csnk2a2 null spermatozoa. Data suggest that CK2α′ deficiency could impair the phosphorylation of nuclear proteins of male germ cells leading to a particular cell‐death pathway characterized by NE protrusions and an unusual pattern of chromatin modifications in spermatids. Mol. Reprod. Dev. 66: 190–201, 2003. © 2003 Wiley‐Liss, Inc.
Association of C‐MYC amplification with progression from the in situ to the invasive stage in C‐MYC‐amplified breast carcinomas
Authors: Robanus‐Maandag Els C , Bosch Cathy AJ , Kristel Petra M , Hart Augustinus AM , Faneyte Ian F , Nederlof Petra M , Peterse Johannes L , van de Vijver Marc J
Published: August 2003

Human carcinoma in situ of the breast already demonstrates genomic changes found in invasive lesions. However, no specific genetic alterations have previously been identified that are associated with progression from the in situ to the invasive stage. By comparative genomic hybridization (CGH) and fluorescence in situ hybridization (FISH) analysis of an invasive breast carcinoma with a large associated in situ component, high‐level amplification of C‐MYC was found in the invasive component only. To determine the frequency of this correlation in a panel of 188 invasive breast carcinomas, 18 additional cases with C‐MYC amplification were identified. Nine of these cases had a detectable adjacent in situ component. FISH analysis demonstrated increased (>5) C‐MYC signals per nucleus in seven invasive components and increased (>4) C‐MYC/centromere 8 signal ratios in five of these. None of the associated in situ components demonstrated these increases. The minimal amplified region was defined at 8q24.13‐8qter. C‐MYC amplification was correlated with overexpression of C‐MYC and two of its target genes, TERT and FBL. Thus, C‐MYC amplification is the first identified genetic alteration that is associated with progression from the in situ to the invasive stage of breast carcinoma. Copyright © 2003 John Wiley & Sons, Ltd.
Adducin expression in cutaneous and oral lesions: α‐ and β‐adducin transcripts down‐regulate with keratinocyte differentiation in stratified epithelia
Authors: Bowen Sharn L , Bloor Balvinder K , Leigh Irene M , Waseem Ahmad
Published: August 2003

Adducin is a heterodimer of α‐ with β‐ or γ‐subunits that regulates the assembly of the spectrin‐based membrane skeleton in erythrocytes. Although adducin has been identified in various non‐erythroid cells and tissues, it has been localized at intercellular junctions only in keratinocytes and epidermis. However, no data are available yet on the regulation of individual adducin genes in differentiating versus hyperproliferating keratinocytes. Due to the unavailability of mono‐specific antibodies for individual adducins, this study has used RT‐PCR and in situ hybridization to investigate the expression of α‐ and β‐adducins in cultured cells and in stratified epithelia including cutaneous and oral lesions. Using RT‐PCR, the α‐transcripts were consistently expressed in all cell lines tested, as well as in normal interfollicular epidermis, whereas the β‐transcripts were more variable and were strongly expressed in K562, A431, and primary keratinocytes. However, in normal skin, oral mucosa, and attached gingivae, the levels of α‐transcripts closely paralleled those for the β‐subunit. In most normal tissues, adducin expression was observed primarily in the proliferating compartments including the basal layer and lower suprabasal layers. Expression of both genes was also up‐regulated in skin diseases characterized by increased cell proliferation and keratinocyte activation, such as basal cell carcinoma (BCC), squamous cell carcinoma (SCC), and psoriasis. It was observed that, in most cases, the expression of both α‐ and β‐adducin was accompanied by increased expression of the proliferation marker Ki‐67 and keratins K6 and K16. Differentiating keratinocytes in normal epithelia as well as in tumours appear to suppress the expression of adducin transcripts. The data suggest that the expression of adducin genes may be linked to cell proliferation and starts to down‐regulate at the onset of differentiation. Copyright © 2003 John Wiley & Sons, Ltd.
Monoclonal antibody 4C4‐mAb specifically recognizes keratan sulphate proteoglycan on human embryonal carcinoma cells
Authors: Fukuma Mariko , Abe Hitoshi , Okita Hajime , Yamada Taketo , Hata Jun‐ichi
Published: August 2003

Germ cell tumours, the most common solid cancers in young males, display pluripotentiality for embryonal and somatic differentiation. Specific surface antigens are useful in the study of cellular differentiation and for clinical diagnosis. A mouse monoclonal antibody (4C4‐mAb) has been developed against a human embryonal carcinoma (EC) cell line (NCR‐G3) isolated from a combined form of testicular germ cell tumour. On immunohistological and immuno‐electron microscopic examination, the 4C4 antigen (4C4) was detected on the surface of NCR‐G3 and gold particles were exclusively detected on the microvilli of the cells. In both formalin‐fixed paraffin wax sections and touch‐smear specimens, 4C4 was detected specifically in EC, while the antigen was not expressed in other types of germ cell tumour or in the other solid tumours tested. Tunicamycin diminished the antigenicity of NCR‐G3 cells. In biochemical studies, 4C4 was found in a high molecular weight region ranging from 1 × 106 to 1 × 107 kD, which disappeared after periodate treatment. The density of 4C4 was 1.5 g/cm3 after equilibrium centrifugation. These results imply that 4C4 is a proteoglycan. Furthermore, endo‐ and exo‐glycosidase treatment revealed that 4C4 is a keratan sulphate proteoglycan that contains sialyl and fucosyl moieties. With EC‐specific and formalin‐resistant characteristics, 4C4 may be a specific marker for diagnosing EC among a variety of germ cell tumours. Copyright © 2003 John Wiley & Sons, Ltd.
Inter‐individual differences in anti‐proteinuric response to ACEi in established adriamycin nephrotic rats are predicted by pretreatment renal damage
Authors: Kramer Andrea B , Laverman Gozewijn D , van Goor Harry , Navis Gerjan
Published: August 2003

ACE inhibition (ACEi) reduces proteinuria and provides reno‐protection, but not all subjects benefit from ACEi. Individual differences in the reduction in proteinuria at the onset of treatment and in residual proteinuria during therapy predict differences in renal outcome. The present study investigated whether individual differences in the anti‐proteinuric efficacy of ACEi are explained by differences in the severity of pretreatment renal structural damage and whether differences in the level of residual proteinuria during therapy are explained by the severity of renal structural damage at that time, in adriamycin nephrosis in the rat. Pretreatment renal structural damage was assessed in biopsies 6 weeks after exposure to adriamycin (2 mg/kg iv). Then ACEi (75 mg/l lisinopril,n= 23) or vehicle (n= 10) was administered; renal biopsies were repeated after stabilization of the anti‐proteinuric response (week 8). Early renal damage (interstitial α‐smooth muscle actin expression and macrophage accumulation) and established lesions [focal glomerulosclerosis (FGS) and interstitial fibrosis] were scored. During ACEi, proteinuria fell from 834 (487–851) mg/24 h pretreatment to 153 (66–265) mg/24 h at week 8 (p< 0.05); FGS stabilized from 27 (4–70) arbitrar units (AU) pretreatment to 26 (4–84) at week 12, whereas the vehicle did not affect proteinuria, resulting in progressive FGS: 18 (10–26) AU pretreatment versus 88 (46–130) at week 12 (p< 0.05). All parameters of pretreatment damage significantly predicted the anti‐proteinuric response. Residual proteinuria during ACEi correlated significantly with renal structural damage parameters at that time. Pretreatment renal damage also predicted renal outcome during extended treatment. Thus, in this experimental setting, in rats with the same renal disorder and the same duration of disease, individual differences in pretreatment renal damage, albeit relatively modest, explain individual differences in renal responsiveness to ACEi. This implies that the limits of the efficacy of ACEi are set by prevalent renal damage. Further studies into the mechanisms of individual resistance to the anti‐proteinuric action of ACEi are needed to develop additive intervention strategies. Copyright © 2003 John Wiley & Sons, Ltd.
Stromal cells of fibrodysplasia ossificans progressiva lesions express smooth muscle lineage markers and the osteogenic transcription factor Runx2/Cbfa‐1: clues to a vascular origin of heterotopic ossification?
Authors: Hegyi Laszlo , Gannon Francis H , Glaser David L , Shore Eileen M , Kaplan Frederick S , Shanahan Catherine M
Published: August 2003

Fibrodysplasia ossificans progressiva (FOP) is a rare heritable genetic disorder, which is characterized pathologically by sporadic episodes of explosive growth of mesenchymal cells in skeletal muscle followed by cellular differentiation to heterotopic bone through an endochondral process. This study examined the histological origin and differentiation state of stromal cells in early FOP lesions and investigated the association between the phenotype of these FOP cells and bone formation. Interestingly, FOP lesional stromal cells were found to display characteristics of the smooth muscle (SM) cell lineage and are therefore potentially of vascular origin. These cells co‐express multiple SM lineage markers along with multiple proteins associated with bone formation including the obligate osteogenic transcription factor Runx2/Cbfa‐1. It is hypothesized that the stromal cells of early FOP lesions may be locally recruited vascular cells or cells of the bone marrow stroma and that these cells maintain the potential (given the correct environmental stimuli) to differentiate along an endochondral ossification pathway. Copyright © 2003 John Wiley & Sons, Ltd.
Infrequent somatic Fas mutations but no evidence of Bcl10 mutations or t(11;18) in primary cutaneous MALT‐type lymphoma
Authors: Grønbæk Kirsten , Ralfkiaer Elisabeth , Kalla Jörg , Skovgaard Gunhild L , Guldberg Per
Published: August 2003

Genetic alterations that allow tumour cells to evade apoptosis have recently been identified as key features of extranodal marginal zone lymphoma of mucosa‐associated lymphoid tissue type (MALT‐type lymphoma). The t(11;18), which produces the putative anti‐apoptotic fusion protein API2‐MALT1, has been identified in a large proportion of extracutaneous MALT‐type lymphomas and a smaller fraction of tumours harbour mutations that inactivate the pro‐apoptotic functions of Fas and Bcl10. The present study has examined the status of these genes in 19 primary cutaneous B‐cell lymphomas (PCBCLs), 12 of which were MALT‐type lymphomas according to the WHO classification. None of the 19 PCBCLs carried the t(11;18) and tumour‐specific Bcl10 alterations were not identified at the genomic level or at the mRNA level. Somatic Fas mutations causing truncation of the Fas receptor were identified in two MALT‐type lymphomas. Both patients with Fas mutant PCBCL exhibited benign conditions of dysregulated lymphoproliferation. One had autoimmune diabetes and rheumatoid arthritis and the other had a 25‐year history of recurrent cutaneous pseudo‐lymphomas. It is suggested that Fas mutation permits the survival and hence the accumulation of autoreactive B cells. This expansion of autoreactive B cells is analogous to the expansion of B cells chronically stimulated by exogenous antigens in the development of MALT‐type lymphoma. Copyright © 2003 John Wiley & Sons, Ltd.
Pathogenese der Rheumatoiden Arthritis: Die pathologische Basis
Authors: Burkhardt Harald , Möller Burkhard
Published: August 2003

Die Integration verschiedener pathogenetischer Aspekte der RA ergibt das komplexe Bild einer Erkrankung, deren individuelle Ausprägung unter multiplen genetischen Einflüssen im zeitlichen Verlauf großen Variationen unterworfen ist (Abb. 5). Initial mag ein exogener Faktor, evtl. ein infektiöses Agens, einen Autoimmunprozess anstoßen, der in seiner T‐Zell‐Antwort in Richtung eines Th1‐Phänotyps polarisiert ist und sich primär gegen gelenkspezifische Antigene wendet. Die sekundäre Ausweitung der Immunantwort im Rahmen eines so genannten “Epitope spreadings” kann auch extraartikuläre Antigene erfassen und im Rahmen einer Immunkomplexvaskulitis zu systemischen Manifestationen führen. Die persistierende Freisetzung proinflammatorischer Zytokine im Gelenk fördert die Transformation der Synovialfibroblasten in einen aggressiven Phänotyp, der schließlich nur noch bedingt parakrinen Regulationsmechanismen unterliegt. Sich verstärkende Rückkopplungsmechanismen auf verschiedenen Ebenen bewirken die Stabilisierung des Prozesses in Richtung auf die gewebedestruierende Endstrecke des chronischen Entzündungsprozesses.
Rekombinante Wirkstoffe bei Rheumatoider Arthritis: Kineret®, Enbrel®, Remicade® und Co.
Authors: Zündorf Ilse , Dingermann Theodor
Published: August 2003

Für die Therapie der Rheumatoiden Arthritis stehen bereits einige sehr potente rekombinante Wirkstoffe zur Verfügung, und das Potenzial an weiteren Wirkstoffoptionen ist bei weitem noch nicht ausgeschöpft. Die genauere Kenntnis der Pathogenese liefert dazu die Grundlagen, und mit den gentechnischen Methoden stehen die Techniken zur Umsetzung zur Verfügung. Allerdings bleiben die Probleme, dass diese Wirkstoffe parenteral zu applizierende und sehr kostenintensive Proteine sind — Probleme, auf die an anderer Stelle in diesem Heft eingegangen wird.
Galenische Formulierung rekombinanter Wirkstoffe: Problem Arzneistoffstabilität
Authors: Zimmer Andreas
Published: August 2003

Die galenische Formulierung von rekombinanten Wirkstoffen zur Behandlung der RA beschränkt sich auf parenteral zu applizierende Darreichungsformen. Es werden kleinvolumige, s. c. zu applizierende Injektionslösungen bis hin zu Infusionslösungen monoklonaler Antikörper eingesetzt. Als Herausforderung gilt es nach wie vor, die Stabilität der Proteine über einen Zeitraum von 2 bis 3 Jahren in der Arzneiform zu gewährleisten. Heute gilt die Lyophilisation als Technik der Wahl, um diesen Anforderungen gerecht zu werden.
Stellenwert rekombinanter Wirkstoffe in der Therapie der RA: Beurteilung der Studien
Authors: Kaltwasser Joachim Peter
Published: August 2003

Sowohl die diagnostischen Möglichkeiten zur sicheren und vor allem frühen Erkennung der Rheumatoiden Arthritis als auch die Kriterien für den therapeutischen Effekt als Grundlage klinischer Arzneimittelprüfungen haben sich in den letzten Jahren erheblich verbessert. Basierend auf diesen besser systematisierten Kriterien sind Verbesserungen in der sinnvollen Auswahl konventioneller Antirheumatika und in der objektiven Bewertung neu entwickelter Arzneimittel erreicht worden. Völlig neue Dimensionen in der Arzneimitteltherapie der Rheumatoiden Arthritis haben die in jüngster Zeit eingeführten rekombinanten Wirkstoffe mit der spezifischen Zielrichtung einer Inhibition proinflammatorischer, bei der RA pathogenetisch relevanter Zytokine eingeführt. Bislang liegen ausgedehnte Erfahrungen aus großen klinischen Studien für die TNF‐α‐inhibierenden monoklonalen Antikörper Infliximab und Adalimumab, das TNF‐α‐Rezeptor‐Fusionsprotein Etanercept und den rekombinant hergestellten, natürlichen IL‐1‐Rezeptorantagonisten Anakinra vor.

Alle vier Wirkstoffe weisen bisher nicht gekannte Effektivitätsdaten auf, die erstmals in relevantem Umfang anhaltende weitgehende Teilremissionen bei RA unter Einschluss des knochendestruierenden Prozesses ermöglichen. Die vorhandene Datenlage erlaubt eine sinnvolle Indikationsabgrenzung für diese neue, sehr vielversprechende Wirkstoffgruppe. Allerdings stehen hohe Arzneimittelpreise — unabhängig von den medizinischen Gesichtspunkten — häufig einer Anwendung noch im Wege.
Gestufte Verfahren zur Behandlung der RA: Wann welches Arzneimittel?
Authors: Bolten Wolfgang W.
Published: August 2003

Die Rheumatoide Arthritis ist eine chronisch entzündliche Systemerkrankung, die mit Schmerzen, Funktionsstörungen, Verminderung der Lebensqualität und Lebenszeitverkürzung einhergeht. Die symptomatische Behandlung erfolgt mit nichtsteroidalen Antirheumatika (NSAR) oder ggf. Paracetamol. Das höhere gastrointestinale Risiko der NSAR kann durch die Kombination mit Protonenpumpenhemmern oder den Austausch des NSAR gegen COX‐2‐selektive Antirheumatika verringert werden. Corticosteroide werden bei entzündlichen Krankheitsschüben und niedrig dosiert bei geeigneten Patienten auch in der Langzeittherapie eingesetzt. Herkömmliche kostengünstige Basistherapeutika (Hydroxychloroquin, Goldsalze, Methotrexat, Leflunomid, Sulfasalazin) werden je nach Krankheitsaktivität, individueller Wirksamkeit und Verträglichkeit einzeln oder in Kombination zur Begrenzung des Krankheitsfortschritts eingesetzt. Wenn Methotrexat und andere Basistherapeutika keine ausreichende Wirkung haben oder unverträglich sind, dann müssen Biologika (TNF‐α‐Antagonisten, IL‐1‐Rezeptor‐Blocker) eingesetzt werden. In der Schwangerschaft reicht meist die symptomatische Behandlung mit NSAR oder Corticosteroiden aus.
Rheumatiker unter der Therapie mit rekombinanten Arzneimitteln: Fragen und Antworten
Authors: Karger Thomas , Missler‐Karger Barbara
Published: August 2003

Die neuen Behandlungsoptionen auf molekularer Ebene („molekulare Chirurgie”︁) eröffnen neue, bisher nie erreichte therapeutische Möglichkeiten. In der Phylogenese haben aber Mechanismen, die wir unter einseitiger Betrachtungsweise als „pathophysiologisch”︁ betrachten, teleologisch eine Bedeutung, die wir nicht kennen. Hätte die Evolution sie sonst entwickelt? Wenn wir derartige — biologisch gesehen sinnvolle — Prozesse isoliert blockieren, sind neue, uns noch nicht bekannte Risiken zu erwarten.

Die wissenschaftliche Fachgesellschaft hat dezidierte Empfehlungen erarbeitet, die klarstellen, wer Indikation und Therapie durchführt (internistische Rheumatologen), welche Qualitäts‐ und Sicherheitskontrollen erforderlich sind (DAS 28, ggf. auch HAQ oder FFbH, Tuberkulose‐Screening, Labor‐ und Röntgenverlaufskontrollen) und somit die Verantwortlichkeit klargestellt. Der Umgang mit rekombinanten Arzneimitteln in der täglichen Praxis erfordert den Spezialisten, der mit den Instrumenten und den Risiken vertraut ist.
Kleine RNA‐Moleküle — ganz groß
Authors: Gießen Hannelore
Published: August 2003

Lange Zeit hat man in der RNA nur einen Vermittler für die Umsetzung des genetischen Codes gesehen. Jetzt mehren sich die Hinweise, dass sie in der Zelle noch ganz andere Aufgaben erfüllt. In den letzten Jahren wurde entdeckt, dass kurze RNA‐Moleküle bei der Zellteilung für Ordnung auf den einzelnen Chromosomen sorgen. Verschiedene Gene kann die small RNA ein‐ und ausschalten und, wenn nötig, Teile des Erbmaterials sogar ganz löschen.
Wie kann man Resistenzen aufheben?
Authors: Holzgrabe Ulrike
Published: August 2003

Antibiotika‐Resistenzen entstehen unter anderem dadurch, dass das Bakterium den Wirkstoff chemisch modifiziert. Prominentestes Beispiel sind die β‐Lactam‐Antibiotika, deren Lactamring mittels der von den Bakterien gebildeten Lactamasen hydrolysiert wird.
Symptomatic postural hypotension in diabetes: aetiology and management
Authors: Winkler AS , Bosman DR
Published: August 2003

This review explores the problem of symptomatic postural hypotension in diabetes, a condition that is fortunately rare but increasingly prevalent in an ageing population. The diverse aetiology and pathophysiology are discussed and relevant investigations and existing therapies are described. New areas of research, which may have therapeutic implications, such as recombinant erythropoietin and octreotide, are also explored. Copyright © 2003 John Wiley & Sons, Ltd.
Cultural aspects of diabetes mellitus in Sudan
Authors: Ahmed AM
Published: August 2003

Diabetes is an increasingly important problem in Sudan, being responsible for 10% of hospital admissions and mortality. This review aims to study the impact of cultural factors on diabetic patients. The impact of the following factors are discussed: fatalism, religious practices, cultural misconceptions, family and gender issues and cultural dietary aspects. Sudan is a multicultural society, with African, Arab, Islamic and Christian roots. Examining the cultural aspects of a common disease such as diabetes is the first step in setting up culture‐sensitive management programmes, and ultimately improving diabetes care. Copyright © 2003 John Wiley & Sons, Ltd.
Capacity building: A primer
Authors: Kinsey David J. , Raker J. Russell
Published: August 2003

The definition and analysis of capacity building in this chapter provides a foundation for understanding how capacity‐building expectations and practices are fulfilled by organizations represented in the rest of this volume. The authors also address organizational change through their discussion of catalytic mechanisms.
Health sector capacity building
Authors: Todsen Dana R.
Published: August 2003

Health sector capacity building is closely connected to the healthy community concept in which people are empowered to improve their individual and collective health by changing the physical and social conditions that directly affect health.
Why capacity building matters and why nonprofits ignore it
Authors: McKinsey & Company
Published: August 2003

Nonprofit organizations often focus organizational development on programs while neglecting systems and infrastructure. Successful capacity building demands attention to strengthening the organization at all levels.
Effect of posture on respiratory function and drive in preterm infants prior to discharge
Authors: Leipälä J.A. , Bhat R. Y. , Rafferty G.F. , Hannam S. , Greenough A.
Published: August 2003

Our objective was to determine the effect of posture on respiratory function and drive in prematurely born infants immediately prior to discharge. Twenty infants (6 oxygen‐dependent), median gestational age 29 weeks (range, 25–32), were studied at a median postconceptional age (PCA) of 36 weeks (range, 33–39). On 2 successive days, infants were studied both supine and prone; each posture was maintained for 3 hr. The order on each day in which postures were studied was randomized between infants. At the end of each 3‐hr period, tidal volume (Vt), inspiratory (Ti) and expiratory (Te) time, respiratory rate, and minute ventilation were measured. In addition, respiratory drive was assessed by measuring the pressure generated in the first 100 msec of an imposed airway occlusion (P0.1), and respiratory muscle strength was assessed by recording the maximum inspiratory pressure (Pimax) generated against an occlusion which was maintained for at least five breaths. Overall, tidal volume was higher (P < 0.05), but respiratory rate (P < 0.05), P0.1 (P < 0.05), and Pimax (P < 0.05) were lower in the prone compared to the supine position. There were no significant differences in Ti or Te between the two postures. In oxygen‐dependent infants only, minute volume was higher in the prone position (P < 0.05). In conclusion, posture‐related differences in respiratory function are present in prematurely born infants studied prior to neonatal unit discharge. Pediatr Pulmonol. 2003; 36:295–300. © 2003 Wiley‐Liss, Inc.
Exhaled nitric oxide corresponds with office evaluation of asthma control
Authors: Meyts Isabelle , Proesmans Marijke , De Boeck Kris
Published: August 2003

Exhaled NO (ENO) has been studied as a noninvasive marker of airway inflammation, and has been shown to be elevated in asthma patients. The aim of this study was to investigate whether ENO measurements differ significantly between groups of asthmatic children with different disease control and to compare ENO measurements with the clinical assessment of asthma control. Seventy‐three children between 5–18 years old with a diagnosis of asthma were recruited. ENO was measured online during a slow vital capacity maneuver. The mean of three plateau NO levels was used for analysis. Baseline and postbronchodilator spirometry were performed. The assessment of disease control was based on the frequency of use of beta2‐agonists, occurrence of day‐ and nighttime asthma symptoms, and spirometry results. Twenty‐one children (group 1) had good asthma control. In 31 patients (group 2), asthma control was acceptable. In 21 patients (group 3), asthma was insufficiently controlled. ENO levels were (median (quartiles)): group 1, 11 ppb (9–21); group 2, 15 ppb (11–26); and group 3, 28 ppb (19–33). Measurements were significantly different between all three groups (P = 0.009, Kruskal‐Wallis), between groups 1 and 3 (P = 0.01, Mann‐Whitney U test), and between groups 2 and 3 (P = 0.01, Mann‐Whitney‐U test). The same was true for reversibility testing. We found significantly different ENO levels between a group of pediatric asthma patients with insufficient and good/sufficient control, as defined by clinical assessment. These results suggest that ENO measurements may be useful for monitoring asthma patients. Pediatr Pulmonol. 2003; 36:283–289. © 2003 Wiley‐Liss, Inc.
Adherence to montelukast versus inhaled corticosteroids in children with asthma
Authors: Carter Edward R. , Ananthakrishnan Madhu
Published: August 2003

Our objective was to compare adherence to montelukast with adherence to inhaled corticosteroids (ICS) in children with persistent asthma. In this retrospective study, we obtained data from a computerized hospital pharmacy database and medical records on 14l patients, 3–18 years of age, who received care for asthma at a military medical center. For each patient, we collected fill/refill data from a consecutive 6‐month period and calculated an adherence rate, i.e., (# doses filled/# doses prescribed) × 100. We then determined whether the patient had “good adherence” (adherence rate, ≥70%) or “very poor adherence” (adherence rate, <50%). Eighty‐one children were prescribed montelukast, and 104 took ICS; 44 of these children took both. The majority of patients had mild or moderate persistent asthma. There was no significant difference between mean adherence rates: 71% (95% CI, 65–77%) for montelukast vs. 67% (95% CI, 61–73%) for ICS. Fifty‐one percent of patients taking montelukast had good adherence, compared to 41% in the ICS group (P = 0.27). Nineteen percent of the montelukast group and 26% of the ICS group had very poor adherence (P = 0.31). Using pharmacy refill data, we found that children with asthma were no more likely to take montelukast than inhaled corticosteroids. Adherence to both medications was suboptimal, even in a system that provides free and easy access to medications. Pediatr Pulmonol. 2003; 36:.301–304. Published 2003 Wiley‐Liss, Inc.
Combination anti‐inflammatory and antiviral therapy of influenza in a cotton rat model
Authors: Ottolini Martin , Blanco Jorge , Porter David , Peterson Lisa , Curtis Spencer , Prince Gregory
Published: August 2003

The cotton rat was evaluated as a model for anti‐inflammatory and antiviral influenza therapy. Beginning 3 days after intranasal infection with 107 tissue culture infectious doses—50% (TCID)50 of an H3N2 human influenza, animals were treated topically via intranasal lavage with a range of doses of triamcinolone acetonide (1, 4, or 16 mg/kg), alone or in combination with a neuraminidase inhibitor or anti‐influenza convalescent serum. Pulmonary histopathologic changes were dramatically decreased in animals treated with 4 or 16 mg/kg of triamcinolone, with little additional benefit from addition of a neuraminidase inhibitor or topical serum, agents which were much less effective when used alone. A high degree of suppression of IFN‐γ levels was observed in all combinations where 4 or 16 mg/kg of triamcinolone were used. Viral replication was not prolonged by corticosteroid therapy. Tissue damage during influenza infection may be greatly reduced by combination antiviral and anti‐inflammatory therapy. Pediatr Pulmonol. 2003; 36:290–294. Published 2003 Wiley‐Liss, Inc.
Rapidly progressing case of Mounier‐Kuhn Syndrome in early childhood
Authors: Hubbard Melissa , Masters I. Brent , Chang Anne B.
Published: August 2003

Reports of Mounier‐Kuhn Syndrome in childhood are extremely rare, as it usually presents in the third or fourth decades. In the only other report of a case diagnosed in early childhood, the child was well at the time of publication. We report on a 15‐month‐old boy who presented with his first respiratory illness at age 3 months. His disease course was more severe than previously reported. Diagnosis was characteristically delayed until acute respiratory failure complicated a respiratory tract infection at 13 months, prompting high‐resolution computerized tomography (HRCT) of the chest. He gradually deteriorated, eventually requiring ventilatory support; death occurred at age 15 months. This report illustrates the clinical heterogeneity of the syndrome. We review the theories about etiology and the recognized clinical findings in adults. Incidence may be higher than previously estimated, and tracheobronchomegaly (TBM) should be considered as a cause of respiratory failure and recurrent pneumonia in children where other investigations, including chest X‐ray (CXR), are normal. A chest HRCT (cHRCT) scan and bronchoscopy are usually diagnostic. Pediatr Pulmonol. 2003; 36:353–356. © 2003 Wiley‐Liss, Inc.
Assessing correlations between spirometry and Shwachman‐Kulczycki score in children and adolescents
Authors: Assis I. , Camargos P.A.M. , Reis F.J.C. , Sulmonett N. , Carneiro A.P.S.
Published: August 2003

A retrospective observational study was carried out to analyze the correlations between formal pulmonary function tests and the Shwachman‐Kulczycki (SK) score. Forty‐six Brazilian cystic fibrosis (CF) patients, clinically stable, aged 7–19 years, were included. Clinical and radiological findings of the SK score system and spirometry variables were assessed by independent observers in a blinded manner. The strongest correlation (r = 0.75; 95% CI, 0.59–0.85; P < 0.001) was found for forced expiratory volume in 1 sec (FEV1). Despite some peculiarities of our patients, the results are in agreement with studies undertaken in industrialized countries, showing good correlation between FEV1 and SK score and its usefulness in managing CF. Accordingly, in countries where pulmonary function testing is not available, the SK score remains a valuable parameter on which to base treatment results. Pediatr Pulmonol. 2003; 36:305–309. © 2003 Wiley‐Liss, Inc.
Extracorporeal life support in pertussis
Authors: Pooboni Suneel , Roberts Neil , Westrope Claire , Jenkins David R. , Killer Hilliary , Pandya Hitesh C. , Firmin Richard K.
Published: August 2003

Severe B. pertussis infection in infants is characterized by severe respiratory failure, pulmonary hypertension, leukocytosis, and death. This retrospective case analysis highlights the course and outcome of severe B. pertussis infection treated with extracorporeal membrane oxygenation (ECMO) at a single center. Over the last decade, out of a total caseload of nearly 800 infants and children, 12 infants with severe B. pertussis have been referred for ECMO therapy to our center. All infants with pertussis infection who received ECMO therapy were less than 3 months of age at presentation and unvaccinated. There was a high mortality rate (7 of 12 infants died), which was associated with an elevated neutrophil count at presentation and multiorgan dysfunction characterized by intractable pulmonary hypertension, persistent systemic hypotension, renal insufficiency, and fits. ECMO should be offered to children with pertussis infection and respiratory failure refractory to mechanical ventilation. However, further research is required to determine the optimal management for infants receiving ECMO therapy with this disease. Pediatr Pulmonol. 2003; 36:310–315. © 2003 Wiley‐Liss, Inc.
Pseudomonas aeruginosa and Burkholderia cepacia cannot be detected by PCR in the breath condensate of patients with cystic fibrosis
Authors: Vogelberg Christian , Hirsch Thomas , Rösen‐Wolff Angela , Kerkmann Marie‐Louise , Leupold Wolfgang
Published: August 2003

The collection of sputum for microbiological examination in young cystic fibrosis patients can be very difficult. However, a knowledge of bacterial flora colonizing the patient's airways is of paramount importance for proper antimicrobial therapy. It is also known that cystic fibrosis patients colonized by Pseudomonas species have a poorer prognosis than Pseudomonas‐negative patients. Noninvasive ways of diagnosing airway inflammation that require only minimal cooperation of the patient might yield new possibilities for early detection of airway colonisation. The breath condensate method as a noninvasive diagnostic technique seems especially appropriate for use in children. Therefore, the aim of this study was to evaluate whether the breath condensate method could be used for detection of Pseudomonas species in children with cystic fibrosis. In total, 32 breath condensate and seven sputum samples were obtained from 13 cystic fibrosis patients with Pseudomonas aeruginosa‐ or Burkholderia cepacia‐positive sputum culture (20 samples were obtained during forced expiration). PCR for combined detection of Pseudomonas aeruginosa, Stenotrophomonas maltophilia, and Burkholderia cepacia was performed. PCR results of all breath condensate samples were negative for Pseudomonas aeruginosa, Stenotrophomonas maltophilia, or Burkholderia cepacia, while all sputum sample results were positive. A minimum DNA quantity of 10 fg could be detected in dilution series of the positive control group. We conclude that the breath condensate method cannot be used as a tool for detection of Pseudomonas species. Pediatr Pulmonol. 2003; 36:348–352. © 2003 Wiley‐Liss, Inc.
An alternative to lung transplantation
Authors: Cohen Gordon , Saglani Sejal , Dinwiddie Robert , Webb Michael , Jaffé Adam
Published: August 2003

Lung volume reduction surgery (LVRS) has been used increasingly in adults for treatment of breathlessness caused by severe emphysema.1 It is of particular benefit to patients with a heterogenous anatomic distribution of emphysema, with obvious target areas for resection,2 as it allows an improved chance of reclaiming function from surrounding compressed lung.3 We report on an 8‐year‐old male with obliterative bronchiolitis in whom LVRS has been used as a measure to significantly improve quality of life and avoid the immediate need for lung transplantation. Pediatr Pulmonol. 2003; 36:357–358. © 2003 Wiley‐Liss, Inc.