bluuebunny/crossref_metadata_2025 huggingface dataset:
Progression of Early Carotid Atherosclerosis Is Only Temporarily Reduced After Antibiotic Treatment of
Chlamydia pneumoniae
Seropositivity
Authors: Sander Dirk , Winbeck Kerstin , Klingelhöfer Jürgen , Etgen Thorleif , Conrad Bastian
Published: February 2004

Background—

Chlamydia pneumoniae
(Cp) infection has been associated with atherosclerosis and cardiovascular events. There are controversial results regarding the beneficial effects of antibiotic therapy on future cardiovascular end points.

Methods and Results—

We determined the long-term effect of a 30-day roxithromycin therapy on intima-to-media thickness (IMT) progression of the common carotid artery in 272 consecutive Cp-positive and Cp-negative patients with ischemic stroke in a prospective, double-blind, randomized trial with a follow-up of 4 years. Cp IgG (≥1:64) or IgA (≥1:16) antibodies were initially found in 125 (46%) patients. During the 3 years before antibiotic therapy, Cp-positive patients showed an enhanced IMT progression even after adjustment for other cardiovascular risk factors (0.12 [0.11 to 0.14] versus 0.07 [0.05 to 0.09] mm/year;
P
<0.005). The 62 Cp-positive patients given roxithromycin showed a reduced IMT progression during the first 2 years compared with the Cp-positive patients without therapy (0.07 [0.045 to 0.095] versus 0.11 [0.088 to 0.132] mm/year;
P
<0.01). However, IMT progression increased again during the third and fourth year to similar values as before treatment. No significant difference in the occurrence of future cardiovascular events was found between both groups during follow-up.

Conclusions—

The only limited positive impact of antibiotic therapy on early atherosclerosis progression in Cp-positive patients observed in our study may explain the negative results of most antibiotic trials on clinical end points.
Familial and Genomic Analyses of Postural Changes in Systolic and Diastolic Blood Pressure
Authors: Harrap Stephen B. , Cui Jisheng S. , Wong Zilla Y. H. , Hopper John L.
Published: February 2004

The physiological adaptation to the erect posture involves integrated neural and cardiovascular responses that might be determined by genetic factors. We examined the familial- and individual-specific components of variance for postural changes in systolic and diastolic blood pressure in 767 volunteer nuclear adult families from the Victorian Family Heart Study. In 274 adult sibling pairs, we made a genome-wide scan using 400 markers for quantitative trait loci linked with the postural changes in systolic and diastolic pressures. Overall, systolic pressure did not change on standing, but there was considerable variation in this phenotype (SD=8.1 mm Hg). Familial analyses revealed that 25% of the variance of change in systolic pressure was attributable to genetic factors. In contrast, diastolic pressure increased by 6.3 mm Hg (SD=7.0 mm Hg) on standing and there was no evidence of contributory genetic factors. Multipoint quantitative genome linkage mapping suggested evidence (
Z
=3.2) of linkage of the postural change in systolic pressure to chromosome 12 but found no genome-wide evidence of linkage for the change in diastolic pressure. These findings suggest that genetic factors determine whether systolic pressure decreases or increases when one stands, possibly as the result of unidentified alleles on chromosome 12. The genetics of postural changes in systolic blood pressure might reflect the general buffering function of the baroreflex; thereby, the predisposition to sudden decreases or increases in systolic pressure might cause postural hypotension or vessel wall disruption, respectively.
Effect of Reduced Angiotensin-Converting Enzyme Gene Expression and Angiotensin-Converting Enzyme Inhibition on Angiotensin and Bradykinin Peptide Levels in Mice
Authors: Campbell Duncan J. , Alexiou Theodora , Xiao Hong D. , Fuchs Sebastien , McKinley Michael J. , Corvol Pierre , Bernstein Kenneth E.
Published: February 2004

There is uncertainty about the contribution of angiotensin-converting enzyme (ACE) to angiotensin II formation, with recent studies suggesting that non-ACE enzymes may be the predominant pathway of angiotensin II formation in kidney, heart, and lung. To investigate the role of ACE in angiotensin II formation, we measured angiotensin I and II levels in blood, kidney, and heart of 2 mouse genetic models (ACE.1 and ACE.4) of reduced somatic ACE gene expression and in blood, kidney, heart, lung, adrenal, and brain of mice administered the ACE inhibitor lisinopril. We also measured the levels of bradykinin (1-9) and its ACE metabolite bradykinin (1-7). Reduced ACE gene expression and ACE inhibition had similar effects on angiotensin and bradykinin peptide levels. Angiotensin II levels were reduced by 70% to 97% in blood, 92% to 99% in kidney, 93% to 99% in heart, 97% in lung, and 85% in adrenal and brain. The marked reductions in angiotensin II/angiotensin I ratio indicated that ACE was responsible for at least 90% of angiotensin I conversion to angiotensin II in blood, kidney, heart, lung, and brain, and at least 77% in adrenal. Blood bradykinin (1-9) levels were increased 6.4-fold to 8.4-fold. Heart bradykinin (1-9) levels were increased in ACE.4 mice and the bradykinin (1-7)/bradykinin (1-9) ratio was reduced in kidney and heart of ACE.4 mice and heart of lisinopril-treated mice. These studies demonstrate that ACE is the predominant pathway of angiotensin II formation in blood and tissues of mice and plays a major role in bradykinin (1-9) metabolism in blood and, to a lesser extent, in kidney and heart.
l
-Arginine Attenuates Hypertension in Pregnant Rats With Reduced Uterine Perfusion Pressure
Authors: Alexander Barbara T. , Llinas Maria T. , Kruckeberg Walter C. , Granger Joey P.
Published: February 2004

A chronic reduction in uterine perfusion pressure in the pregnant rat is associated with significant elevations in mean arterial pressure, proteinuria, and reductions in kidney function as is chronic nitric oxide blockade, suggesting that nitric oxide deficiency may contribute to the clinical manifestations of preeclampsia. The purpose of this study was to determine whether supplementation with
l
-arginine, the precursor for nitric oxide, attenuates the hypertension produced in response to a chronic reduction in uterine perfusion pressure in the pregnant rat. Reduced uterine perfusion was initiated at day 14 of gestation with arterial pressure determined at day 19 of gestation in conscious, chronically instrumented rats. Arterial pressure was significantly elevated in pregnant rats with chronic reductions in uterine perfusion as compared with pregnant control rats (132±2 versus 109±2 mm Hg,
P
<0.01, respectively). Treatment with
l
-arginine (2%) in the drinking water was initiated at day 10 of gestation.
l
-arginine supplementation resulted in a significant decrease in arterial pressure in both pregnant rats with reduced uterine perfusion pressure (113±2 mm Hg treated,
P
<0.01 versus untreated pregnant with reduced uterine perfusion pressure) and pregnant control (97±3 mm Hg treated,
P
<0.01 versus untreated pregnant) rats. However, supplementation with
l
-arginine decreased blood pressure by 19 mm Hg in pregnant with reduced uterine perfusion pressure (untreated versus treated) as compared with 12 mm Hg in pregnant (untreated versus treated) rats. Thus, these results suggest that
l
-arginine supplementation may be beneficial in attenuating the hypertension in preeclampsia.
Estimated Global Transmural Distribution of Activation Rate and Conduction Block During Porcine and Canine Ventricular Fibrillation
Authors: Newton Jonathan C. , Smith William M. , Ideker Raymond E.
Published: February 2004

We quantified ventricular fibrillation (VF) activation rate, conduction block, and organization transmurally in pigs and dogs, whose transmural Purkinje distribution differ. In six pigs and five dogs, 75 to 100 plunge needles, containing four electrodes for the right ventricle (RV) and six electrodes for the left ventricle (LV) and septum, were inserted in vivo. Six VF episodes were electrically initiated and allowed to last for 47 to 180 seconds. From the FFT power spectra, dominant frequency (DF), an estimate of activation rate, and incidence of double peaks (DPI), an estimate of conduction block, were calculated every 8 ms at each electrode. DF was highest at the epicardium and lowest at the endocardium, whereas DPI was highest at the endocardium and lowest at the epicardium for the entire LV and the RV base in both pigs and dogs for the first 70 seconds of VF. This distribution changed little throughout the first 3 minutes of VF in pigs but reversed in dogs by 2 minutes of VF. In conclusion, estimated activation rates and conduction block incidence during VF are not uniformly distributed transmurally. During the first minute of VF, the faster activating LV base epicardium exhibits less estimated block than the slower endocardium, raising the possibility that faster activating epicardium generates wavefronts that drive the endocardium early during VF. Constancy of this pattern in pigs but its reversal by 2 minutes in dogs is consistent with the hypothesis that activation during later VF is driven by Purkinje fibers.
Regeneration of the Infarcted Heart With Stem Cells Derived by Nuclear Transplantation
Authors: Lanza Robert , Moore Malcolm A.S. , Wakayama Teruhiko , Perry Anthony C.F. , Shieh Jae-Hung , Hendrikx Jan , Leri Annarosa , Chimenti Stefano , Monsen Alyssa , Nurzynska Daria , West Michael D. , Kajstura Jan , Anversa Piero
Published: February 2004

Nuclear transfer techniques have been proposed as a strategy for generating an unlimited supply of rejuvenated and histocompatible stem cells for the treatment of cardiac diseases. For this purpose, c-kit–positive fetal liver stem cells obtained from cloned embryos were injected in the border zone of infarcted mice to induce tissue reconstitution. Cloned embryos were derived from somatic cell fusion between nuclei of cultured LacZ-positive fibroblasts and enucleated oocytes of a different mouse strain. We report that regenerating myocardium replaced 38% of the scar at 1 month. The rebuilt tissue expressed LacZ and was composed of myocytes and vessels connected with the coronary circulation. Myocytes were functionally competent and expressed contractile proteins, desmin, connexin43, and N-cadherin. These structural characteristics indicated that the new myocytes were electrically and mechanically coupled. Similarly, the formed coronary arterioles and capillary structures contained blood and contributed, therefore, to tissue oxygenation. Cardiac replacement resulted in an improvement of ventricular hemodynamics and in a reduction of diastolic wall stress. These beneficial effects were obtained by stem cell transdifferentiation and commitment to the cardiac cell lineages. Myocardial growth was independent from fusion of the injected stem cells with preexisting partner cells. In conclusion, c-kit–positive stem cells derived by nuclear transfer cloning restore infarcted myocardium. Although problems currently plague nuclear transplantation, including the potential for epigenetic and imprinting abnormalities, stem cells derived from cloned embryos are sufficiently normal to repair damaged tissue in vivo. Importantly, the magnitude of myocardial regeneration obtained in this study is significantly superior to that achieved with adult bone marrow cells.
Ten-Year Risk of First Recurrent Stroke and Disability After First-Ever Stroke in the Perth Community Stroke Study
Authors: Hardie Kate , Hankey Graeme J. , Jamrozik Konrad , Broadhurst Robyn J. , Anderson Craig
Published: February 2004

Background and Purpose—

Limited information exists on the long-term prognosis after first-ever stroke. We aimed to determine the absolute frequency of first recurrent stroke and disability and the relative frequency of recurrent stroke over 10 years after first-ever stroke in Perth, Western Australia.

Methods—

For a 12-month period beginning February 1989, all individuals with suspected acute stroke or transient ischemic attack who lived in a geographically defined and representative region of Perth were registered prospectively. Patients with a definite first-ever stroke were followed up 10 years after the index event.

Results—

Over 10 years of follow-up, the cumulative risk of a first recurrent stroke was 43% (95% confidence interval [CI], 34 to 51). After the first year after first-ever stroke, the average annual risk of recurrent stroke was ≈4%. Case fatality at 30 days after first recurrent stroke was 41%, which was significantly greater than the case fatality at 30 days after first-ever stroke (22%) (
P
=0.003). For 30-day survivors of first-ever stroke, the 10-year cumulative risk of death or new institutionalization was 79% (95% CI, 73 to 85) and of death or new disability was 87% (95% CI, 81 to 92).

Conclusions—

Over 10 years of follow-up, the risk of first recurrent stroke is 6 times greater than the risk of first-ever stroke in the general population of the same age and sex, almost one half of survivors remain disabled, and one seventh require institutional care. Effective strategies for prevention of stroke need to be implemented early, monitored frequently, and maintained long term after first-ever stroke.
Influence of Gender on K
+
-Induced Cerebral Vasodilatation
Authors: Chrissobolis Sophocles , Sobey Christopher G.
Published: February 2004

Background and Purpose—

It is not known whether cerebral vasoprotective mechanisms in females include increased function of arterial K
+
channels. We hypothesized that vasodilator responses mediated by activation of inwardly rectifying K
+
(K
IR
) channels are greater in cerebral arteries of female versus male rats and that this is due to the effects of estrogen.

Methods—

Changes in basilar artery diameter were measured with a cranial window preparation in anesthetized Sprague-Dawley rats.

Results—

K
+
(5 and 10 mmol/L) caused greater vasodilatation in females (percent maximum, 21±3% and 58±7%, respectively) versus males (11±1% and 37±4%, respectively;
P
<0.05). In contrast, vasodilator responses to aprikalim (1 and 3 μmol/L) or acetylcholine (ACh, 1 and 10 μmol/L) did not differ between the genders. The selective K
IR
channel inhibitor barium ion (30 μmol/L) decreased basilar artery diameter in males but not females (−7±1% versus −2±1%,
P
<0.05) and selectively inhibited K
+
-induced vasodilatation by ≈50% in both groups. Ovariectomy of female rats resulted in smaller vasodilator effects of K
+
, and chronic treatment of these rats with 17β-estradiol (0.01 mg/kg per day for 7 days) normalized K+-induced vasodilatation. Furthermore, the selective M2 muscarinic ACh receptor antagonist methoctramine (1 μmol/L) increased responses to K
+
in males to levels equivalent to responses in females but had no effect on responses to K
+
in females.

Conclusions—

K
+
is a more powerful vasodilator in the female versus male cerebral circulation. This difference is estrogen dependent and could be due to a lack of M2 muscarinic ACh receptor–induced inhibition of K
IR
channel activation by K
+
in female cerebral arteries.
Risk of Intracerebral Hemorrhage in Patients With Arterial Versus Cardiac Origin of Cerebral Ischemia on Aspirin or Placebo
Authors: Ariesen M.J. , Algra A. , Koudstaal P.J. , Rothwell P.M. , van Walraven C.
Published: February 2004

Background and Purpose—

Patients who are anticoagulated after cerebral ischemia have a 19-fold-higher risk of intracerebral hemorrhage (ICH) if they had an arterial rather than a cardiac source. To determine whether this excess risk of ICH was due to the underlying disease (cerebral ischemia of arterial versus cardiac origin) or whether it depended on the antithrombotic regimen, we studied the risk of ICH in arterial versus cardiac origin of cerebral ischemia in patients who received aspirin or no antithrombotic drugs.

Methods—

Individual patient data of patients who received aspirin or placebo after cerebral ischemia were obtained from 9 clinical trials. Presence of atrial fibrillation was considered evidence of a cardiac source. Otherwise, events were considered of arterial origin. Cox proportional-hazards modeling was used for univariate and multivariate analyses.

Results—

Fifty-four ICHs occurred in 16 625 patient-years in the aspirin-treated patients, and 7 ICHs occurred in 4317 patient-years in those on placebo. After multivariate adjustment for age, sex, current smoking, history of hypertension and diabetes, and aspirin dose (aspirin-treated patients only), the hazard ratio for ICH in patients with an arterial versus a cardiac source was 0.74 (95% confidence interval, 0.30 to 1.82) for aspirin-treated patients and 4.34 (95% confidence interval, 0.35 to 54) for placebo-randomized patients.

Conclusions—

Our findings do not confirm the previous finding of an excess risk of ICH in patients with cerebral ischemia of arterial origin. Therefore, it seems that having cerebral ischemia of arterial origin by itself is not associated with an increased risk of ICH, but only in combination with high-intensity anticoagulation.
Influence of incubation conditions on hatching success, embryo development and hatchling phenotype of common chameleon (Chamaeleo chamaeleon) eggs
Authors: Díaz-Paniagua  , Cuadrado
Published: February 2004

Chameleons lay flexible-shelled eggs that are affected by environmental characteristics during incubation. We incubated eggs of six clutches under four treatments using two different constant temperatures (25 and 29°C) and water potentials (-150 kPa and-600 kPa). Eggs gained mass in all treatments, especially under the wetter and warmer conditions. Temperature, not water potential, affected hatching rate, which was 100% under 25°C and 64% under 29°C. Development time was considered as the period during which eggs were incubated under both constant temperatures and differed between treatments, from an average length of 98.8 days in the wettest and warmest media to 144.2 days in the driest and coldest one. Differences in hatchling phenotype were mostly explained by clutch identity, although it was also affected by incubation treatments, with bigger and heavier hatchlings in colder treatments. Incubation conditions did not affect hatchling growth nor survival rates, but survivors were differentiated by their higher increase in egg mass during incubation. Los huevos de los camaleones tienen la cáscara flexible, estando influidos por las características del medio en que se encuentran. Se incubaron huevos de seis puestas en cuatro tratamientos, usando dos temperaturas de incubación constantes (25 y 29°C) y dos potenciales hídricos (-150 kPa y-600 kPa). Los huevos incrementaron de peso en todos los tratamientos, principalmente en los de condiciones más húmedas y cálidas. La tasa de eclosión resultó influida por el efecto de la temperatura, pero no por el del potencial hídrico. Eclosionó el 100% de los huevos incubados a 25°C y el 64% de los incubados a 29°C. Se apreciaron diferencias en la duración del desarrollo entre los huevos incubados en los distintos tratamientos, variando el promedio entre 98.8 días (en las condiciones más húmedas y cálidas) y 144.2 días (en el más seco y frío). Las diferencias apreciadas en el fenotipo de las crías se atribuyen principalmente a su origen materno, aunque también se apreciaron influencias de las condiciones de incubación, resultando crías de mayor tamaño y peso en los tratamientos de menor temperatura. Las condiciones de incubación no afectaron al crecimiento ni a la supervivencia de las crías, sobreviviendo las que procedían de huevos con mayor incremento de peso durante la incubación.
Aspects of population ecology of Testudo hermanni hermanni from Asinara Island, NW Sardinia (Italy, Western Mediterranean Sea): preliminary data
Authors: Zuffi Marco , Corti Claudia
Published: February 2004

Hermann's tortoise (Testudo hermanni Gmelin) of the Island of Asinara (NW Sardinia, Italy) has been studied during a period of four seasons. This population consisted of several individuals of large body size, some of them being amongst the largest known for this species. Bony shells of males were commonly found on this island, while the same was not true for female bony shells. Although wild boars (Sus scrofa) are potential large sized predators for tortoises, the analysis of carapace morphology did not reveal injuries signs due to bites. Male tortoises could die due to up-turning, as the result of male-male combats during the mating season, to possible hyperthermia when upturned, and to predation carried out by birds particularly when upturned. Estimated density was 4.88 individuals ha-1, which was similar to that of conspecifics from other areas studied so far.
Giant and long-lived? Age structure in Macroscincus coctei, an extinct skink from Cape Verde
Authors: Guarino Fabio , Andreone Franco
Published: February 2004

The age structure of the extinct giant Cape Verde skink Macroscincus coctei was studied using skeletochronology applied to 11 preserved males, 15 females, and one juvenile, collected at the end of the 19th century. Counts of lines of arrested growth (LAGs) indicated that in this species — which lived on islets with extreme natural conditions (e.g., hard aridity, lack of rainfall) — the maximum life span was estimated as 16 years for males and 12 years for females; the single juvenile was 4 years old. Although the sexes had similar mean ages, mean body length of males was significantly higher than that of females (283 versus 255 mm). There was no clear relationship between body length and estimated age, although the largest males were also the oldest. The analysis of LAGs revealed a sexual difference in growth rates, males reaching greater size earlier than females.
Aseasonal reproduction and high fecundity in the Cape grass lizard, Cordylus anguinus, in a fire-prone habitat
Authors: du Toit Annemarie , Mouton P. le Fras , Flemming Alexander
Published: February 2004

We investigated life history characters of the Cape grass lizard, Cordylus anguinus, and relate them to survival in the fire-prone habitat in which it occurs. Unlike in other cordylids, reproductive activity was found to be asynchronous among females, with vitellogenic and gravid females encountered virtually throughout the year. Aseasonal breeding will circumvent reduction or complete loss of reproductive effort for any given year due to fire. Female grass-lizards attain significantly larger body sizes than males. Clutch size ranged from three to seventeen and was positively correlated with snout-vent length. Maximum clutch size is more than three times that recorded for any other cordylid. We suggest that high fecundity will allow quick recruitment after a fire. To determine the reproductive cycle exhibited by males, testicular volume and seminiferous tubule diameter were measured, and spermatogenic activity assessed qualitatively, using Licht's classification system. Our data indicate that C. anguinus has a post-nuptial spermatogenic cycle. The species differs from other cordylids having a post-nuptial cycle, in that spermatogenesis already commences in spring.
Distribution of genetic variation and taxonomy of insular and mainland populations of the Italian wall lizard, Podarcis sicula
Authors: Capula Massimo , Ceccarelli Arianna
Published: February 2004

The genetic structure and heterogeneity of Podarcis sicula (Reptilia, Lacertidae) was studied in insular (Pontine Archipelago) and mainland (central and southern Italy) populations by means of allozyme electrophoresis at 20 presumptive gene loci. Genetic variability in the species is low and genetic subdivision is high. The highest values of percent polymorphism and heterozygosity were found in the samples from the southernmost part of Italy (Calabria). The insular samples from the Pontine Archipelago were characterized by loss of alleles and erosion of genetic variability. Population heterogeneity analysis carried out by the estimation of Wright's F-statistics demonstrated substantial genetic differentiation among populations. F-statistics and genetic distance data show that genetic variation is distributed into three population groups. The first group includes the genetically very similar populations from central Italy and the Pontine Archipelago, the second includes the populations from Campania (southern Italy), the third comprises the populations from the southernmost part of the Italian Peninsula (Calabria). Based on the results of the allozyme data, the systematic status of the subspecies of P. sicula occurring in the studied areas is discussed.
Seasonal and daily activity patterns of Vipera latastei in northern Portugal
Authors: Brito José
Published: February 2004

The annual and daily activity cycles of Vipera latastei were investigated in northern Portugal from 1999 to 2001, based on field observations, radio-tracking data and road killed snakes. Annual activity was extended from early March to late October, although some snakes were active during winter. Two annual peaks of activity were observed, a minor one in March-May and a major one in September-October. The daily activity pattern of vipers in spring and autumn was unimodal, whereas in summer they expanded their activity from early morning to late afternoon, and were also active at night. Activity and body exposure of adult males were correlated with meteorological variables. Females had shedding periods in June and July, whereas males shed their skin more frequently in June and August. Sperm was detected in the vas deferens of males in October to December. Females ovulated in June and new-born vipers were found from early August. Mating season is apparently restricted to autumn, and spring mating is either absent or very reduced.
Do Male Black-capped Chickadees Eavesdrop on Song Contests? A Multi-speaker Playback Experiment
Authors: Ratcliffe Laurene , Mennill Daniel
Published: February 2004

Within a network of communicating individuals, animals may gather information about the relative quality of conspecifics by eavesdropping on their signalling interactions. For territorial male songbirds, eavesdropping may be a low-cost, low-risk method for assessing the relative quality of the males around them. We used a three-speaker playback design to evaluate whether male black-capped chickadees (Poecile atricapillus) respond differently to two simulated countersinging intruders who differ only in relative features of their singing performance. We arranged three loudspeakers in an equilateral triangle at the center of playback subjects' territories. After luring males to the first loudspeaker by broadcasting non-song vocalizations, we played songs from the remaining loudspeakers to simulate a countersinging interaction between two male intruders. During the interactions, one simulated intruder consistently overlapped the songs of the other, a behaviour thought to be a signal of directed aggression in songbirds. Territorial male chickadees discriminated between the simulated intruders by preferentially approaching the loudspeaker broadcasting the overlapping signal, suggesting that males eavesdrop on other males' countersinging interactions. Male responses to playback support the idea that overlapping is a more threatening signal than being overlapped. Responses varied with the dominance status of the subject. High-ranking males approached the overlapping loudspeaker in 15 of 16 cases whereas low-ranking males approached the overlapping speaker in only 5 of 10 cases, suggesting that males of different quality may use different tactics for territorial defense.
East Timor: A Critique of the Model of Accountability for Serious Human Rights and International Humanitarian Law Violations
Authors: Othman Mohamed
Published: February 2004

The article examines the model of accountability for international humanitarian law violations committed in East Timor in 1999, in which the Security Council opted for parallel accountability,in East Timor and Indonesia. In the former, it is anchored on a 'mixed' judicial process, administered by 'Special Panels' of the Dili District Court composed of national and international judges. Although its legal framework is sound, and persons 'most responsible' have been indicted, the allocated resources were meager, desired full accountability overambitious, and most accused remain at large. In Indonesia, the ad hoc Human Rights Court's framework is laden withjurisdictional loopholes, and the conduct of prosecutions non-diligent, reflecting 'unwillingness or inability', genuinely to account. If that process had been under the ICC regime, it would have been a valid ground for seizure of jurisdiction by that court. The net result of the model is a misjoinder of human rights perpetrators.
Kostmann syndrome: severe congenital neutropenia associated with defective expression of Bcl-2, constitutive mitochondrial release of cytochrome c, and excessive apoptosis of myeloid progenitor cells
Authors: Carlsson Göran , Aprikyan Andrew A. G. , Tehranchi Ramin , Dale David C. , Porwit Anna , Hellström-Lindberg Eva , Palmblad Jan , Henter Jan-Inge , Fadeel Bengt
Published: February 2004

Kostmann syndrome, or severe congenital neutropenia (SCN), is an autosomal recessive disorder of neutrophil production. To investigate the potential role of apoptosis in SCN, bone marrow aspirates and biopsies were obtained from 4 patients belonging to the kindred originally described by Kostmann and 1 patient with SCN of unknown inheritance. An elevated degree of apoptosis was observed in the bone marrow of these patients, and a selective decrease in B-cell lymphoma-2 (Bcl-2) expression was seen in myeloid progenitor cells. Furthermore, in vitro apoptosis of bone marrow-derived Kostmann progenitor cells was increased, and mitochondrial release of cytochrome c was detected in CD34+ and CD33+ progenitors from patients, but not in controls. Administration of granulocyte colony-stimulating factor (G-CSF) restored Bcl-2 expression and improved survival of myeloid progenitor cells. In addition, cytochrome c release was partially reversed upon incubation of progenitor cells with G-CSF. In sum, these studies establish a role for mitochondria-dependent apoptosis in the pathogenesis of Kostmann syndrome and yield a tentative explanation for the beneficial effect of growth factor administration in these patients. (Blood. 2004;103:3355-3361)
Reconstitution of dendritic and natural killer–cell subsets after allogeneic stem cell transplantation: effects of endogenous flt3 ligand
Authors: Chklovskaia Elena , Nowbakht Pegah , Nissen Catherine , Gratwohl Alois , Bargetzi Mario , Wodnar-Filipowicz Aleksandra
Published: February 2004

Recovery of dendritic cells (DCs) and natural killer (NK) cells after allogeneic stem cell transplantation (SCT) is important for allograft responses and antitumor immunity and thus for treatment outcome. Regulation of this regenerative process is not well understood. We investigated the influence of endogenous cytokines on the recovery and diversification of DC and NK cell subsets up to 6 months after SCT. Reconstitution of circulating DCs and NK cells was rapid but accompanied by prolonged skewing of cell subsets. The speed of recovery of CD11c+CD123low DC1 exceeded that of CD11c– CD123+ DC2, and correlated with plasma levels of flt3 ligand (FL), but not with granulocyte or granulocyte-macrophage colony-stimulating factors and stem cell factor. There was a 5-fold increase in interferon-γ–producing CD56highCD16–/low NK cells and a corresponding reduction in the CD56lowCD16high subset, accompanied by strongly reduced NK cell cytotoxicity. In vitro data implicate an inhibitory effect of cyclosporin A on NK cell differentiation and cytotoxicity. NK cell numbers did not correlate with plasma levels of FL or interleukin 15. Our results demonstrate that endogenous FL has distinct effects on the kinetics of reconstitution of DCs and NK cells and have potential implications for the modulation of immune responses after allogeneic SCT.
Angiopoietin-related growth factor (AGF) promotes angiogenesis
Authors: Oike Yuichi , Ito Yasuhiro , Maekawa Hiromitsu , Morisada Tohru , Kubota Yoshiaki , Akao Masaki , Urano Takashi , Yasunaga Kunio , Suda Toshio
Published: February 2004

We report here the identification of angiopoietin-related growth factor (AGF) as a positive mediator for angiogenesis. To investigate the biologic function of AGF in angiogenesis, we analyzed the vasculature in the dermis of transgenic mice expressing AGF in mouse epidermal keratinocytes (K14-AGF). K14-AGF transgenic mice were grossly red, especially in the ears and snout, suggesting that hypervascularization had occurred in their skin. Histologic examination of ear skin from K14-AGF transgenic mice revealed increased numbers of microvessels in the dermis, whereas the expression of several angiogenic factors, such as basic fibroblast growth factor (bFGF), vascular endothelial growth factors (VEGFs), and angiopoietin-1 (Ang-1), was decreased. We showed that AGF is a secreted protein and does not bind to tyrosine kinase with immunoglobulin and EGF-homology domain (Tie1) or Tie2 receptors. An in vitro chamber assay revealed that AGF directly promotes chemotactic activity of vascular endothelial cells. Both mouse corneal and matrigel plug assays showed that AGF induces neovascularization in vivo. Furthermore, we found that plasma leakage occurred after direct injection of AGF into the mouse dermis, suggesting that AGF directly induces a permeability change in the local vasculature. On the basis of these observations, we propose that AGF is a novel angiogenic factor and that handling of its biologic functions could lead to novel therapeutic strategies for control of angiogenesis.
Chimeric CD19 antibody mediates cytotoxic activity against leukemic blasts with effector cells from pediatric patients who received T-cell–depleted allografts
Authors: Lang Peter , Barbin Karin , Feuchtinger Tobias , Greil Johann , Peipp Matthias , Zunino Susan J. , Pfeiffer Matthias , Handgretinger Rupert , Niethammer Dietrich , Fey Georg H.
Published: February 2004

Relapse is a major problem after transplantation in children with acute B-lineage leukemias, and new therapies are needed to increase graft-versus-leukemia (GvL) effects without inducing graft-versus-host disease (GvHD). Here, we studied the ability of effector cells recovered from patients after transplantation with positive-selected stem cells from alternative donors to induce antibody-dependent cellular cytotoxicity (ADCC). For this purpose, a chimeric CD19 antibody, CD19-4G7chim, was generated. This antibody efficiently mediated ADCC against primary acute lymphoblastic leukemia (ALL) blasts by using purified natural killer (NK) cells from healthy donors or mononuclear cells from patients as effector cells. Increased lysis was obtained after stimulation of effector cells with interleukin-2 (IL-2). ADCC was not prevented by inhibitory effects mediated by HLA class I. We propose that treatment with chimeric CD19 antibodies leading to ADCC by donor-derived NK cells may become a therapeutic option for the post-transplantation treatment of minimal residual B-lineage ALLs.
Pim-1 kinase inhibits STAT5-dependent transcription via its interactions with SOCS1 and SOCS3
Authors: Peltola Katriina J. , Paukku Kirsi , Aho Teija L. T. , Ruuska Marja , Silvennoinen Olli , Koskinen Päivi J.
Published: February 2004

Signal transducer and activator of transcription 5 (STAT5) plays a critical role in cytokine-induced survival of hematopoietic cells. One of the STAT5 target genes is pim-1, which encodes an oncogenic serine/threonine kinase. Here we demonstrate that Pim-1 inhibits STAT5-dependent transcription in cells responsive to interleukin-3, prolactin, or erythropoietin. Ectopic expression of Pim-1 in cytokine-dependent FDCP1 myeloid cells results in reduced tyrosine phosphorylation and DNA binding of STAT5, indicating that Pim-1 interferes already with the initial steps of STAT5 activation. However, the Pim-1 kinase does not directly phosphorylate or bind to STAT5. By contrast, Pim-1 interacts with suppressor of cytokine signaling 1 (SOCS1) and SOCS3 and potentiates their inhibitory effects on STAT5, most likely via phosphorylation-mediated stabilization of the SOCS proteins. Thus, both Pim and SOCS family proteins may be components of a negative feedback mechanism that allows STAT5 to attenuate its own activity.
Dendritic cell–activated CD44hiCD8+ T cells are defective in mediating acute graft-versus-host disease but retain graft-versus-leukemia activity
Authors: Zhang Yi , Joe Gerard , Zhu Jiang , Carroll Richard , Levine Bruce , Hexner Elizabeth , June Carl , Emerson Stephen G.
Published: February 2004

Graft versus host disease (GVHD) is triggered by host antigen-presenting cells (APCs) that activate donor T cells to proliferate and differentiate, but which APC-activated donor T-cell subsets mediate GVHD versus beneficial antitumor effects is not known. Using a CD8+ T cell–dependent mouse model of human GVHD, we found that host dendritic cell (DC)–induced CD44hiCD8+ effector/memory T cells were functionally defective in inducing GVHD, whereas CD44loCD8+ naive phenotype T cells were extremely potent GVHD inducers. Depletion of CD44loCD8+ T cells from host DC-stimulated T cells before transplantation prevented GVHD without impairing their antitumor activity in vivo. Compared with CD44loCD8+ T cells, CD44hiCD8+ T cells expressed high levels of Fas and were efficiently deleted in vivo following transplantation. These results suggest that ex vivo allogeneic DC stimulation of donor CD8+ T cells may be useful for the prevention of GVHD and for optimizing antitumor therapies in vivo.
Regulatory elements of the EKLF gene that direct erythroid cell-specific expression during mammalian development
Authors: Xue Li , Chen Xiaoyong , Chang Yanjie , Bieker James J.
Published: February 2004

Erythroid Krüppel-like factor (EKLF) plays an essential role in enabling β-globin expression during erythroid ontogeny. It is first expressed in the extraembryonic mesoderm of the yolk sac within the morphologically unique cells that give rise to the blood islands, and then later within the hepatic primordia. The BMP4/Smad pathway plays a critical role in the induction of EKLF, and transient transfection analyses demonstrate that sequences located within less than 1 kb of its transcription initiation site are sufficient for high-level erythroid-specific transcription. We have used transgenic analyses to verify that 950 bp located adjacent to the EKLF start site of transcription is sufficient to generate lacZ expression within the blood islands as well as the fetal liver during embryonic development. Of particular importance are 3 regions, 2 of which overlap endogenous erythroid-specific DNase hypersensitive sites, and 1 of which includes the proximal promoter region. The onset of transgene expression mimics that of endogenous EKLF as it begins by day 7.5 (d7.5) to d8.0. In addition, it exhibits a strict hematopoietic specificity, localized only to these cells and not to the adjacent vasculature at all stages examined. Finally, expression is heterocellular, implying that although these elements are sufficient for tissue-specific expression, they do not shield against the position effects of adjacent chromatin. These analyses demonstrate that a surprisingly small DNA segment contains all the information needed to target a linked gene to the hematopoietic compartment at both early and later stages of development, and may be a useful cassette for this purpose.
Clonotypic IgM V/D/J sequence analysis in Waldenstrom macroglobulinemia suggests an unusual B-cell origin and an expansion of polyclonal B cells in peripheral blood
Authors: Kriangkum Jitra , Taylor Brian J. , Treon Steven P. , Mant Michael J. , Belch Andrew R. , Pilarski Linda M.
Published: February 2004

Analysis of clonotypic immunoglobulin M (IgM) from 15 patients with Waldenstrom macroglobulinemia (WM) showed a strong preferential use of the VH3/JH4 gene families. Identification of the WM IgM V/D/J was validated using single-cell analysis, confirming its presence in most B cells. Despite the extensive hypermutated VH genes in 13 of 15 patients, statistical analysis of framework/complementary-determining region (FR/CDR) mutation patterns suggests that they might have escaped antigenic selection. Neither intraclonal diversity nor isotype switching was detectable. Membranous and secreted forms of clonotypic IgM transcripts were present in bone marrow and blood. Single-cell analysis showed that clonotypic B cells coexpress CD20, surface IgM (sIgM), and sIgD but that they lack CD138. Most B cells lacked memory marker CD27 despite their hypermutated variable regions otherwise suggestive of memory status. At diagnosis, circulating B cells in WM are largely clonotypic. However, when monoclonal IgM levels are decreased, clonotypic frequencies are substantially reduced despite elevated CD20+ cells, shown to be polyclonal by DNA sequencing and CDR3 fragment analysis. Thus, WM includes the expansion of circulating, polyclonal B cells. Overall, this work suggests that WM may originate from a largely VH3-restricted, somatically mutated, predominantly CD27-IgM+IgD+ population that cannot undergo class switching, suggestive of B cells that might have bypassed the germinal center. (Blood. 2004;104:2134-2142)
T lymphocytes of recipient origin may contribute to the recovery of specific immune response toward viruses and fungi in children undergoing cord blood transplantation
Authors: Montagna Daniela , Locatelli Franco , Moretta Antonia , Lisini Daniela , Previderè Carlo , Grignani Pierangela , DeStefano Piero , Giorgiani Giovanna , Montini Enrica , Pagani Sara , Comoli Patrizia , Maccario Rita
Published: February 2004

Patients undergoing allogeneic cord blood transplantation (CBT) benefit from a low risk of graft-versus-host disease (GVHD), but there are still concerns that they be able to recover an effective immune capacity early after transplantation. We investigated the ability to develop in vitro T-lymphocyte-mediated immune response toward human cytomegalovirus and Candida albicans antigens, early and late after transplantation, in children given cord blood transplants from either an HLA-identical sibling or an unrelated donor. Proliferative capacity and frequency of antigen-specific T cells were evaluated; antigen-specific CD4+ T-cell clones were also generated and characterized for T-cell receptor repertoire diversity, cytokine phenotype, and their origin (either from donor or patient). We found that the majority of recipients can develop a specific response to viral or fungal antigens already early after transplantation. Antigen-specific T-cell clones of both donor and recipient origin contributed to the reconstitution of immune response. Antigen-specific T lymphocytes of recipient origin were detected in patients receiving a transplant from a relative, after a chemotherapy-based conditioning regimen, and who did not have GVHD. Our results document, at a clonal level, that after CBT recovery of either polyclonal or pauciclonal T-cell response toward widespread pathogens is prompt, with some patients benefiting from a contribution of recipient-derived cells. (Blood. 2004;103:4322-4329)
MuTrack: a genome analysis system for large-scale mutagenesis in the mouse
Authors: Baker Erich J , Galloway Leslie , Jackson Barbara , Schmoyer Denise , Snoddy Jay
Published: February 2004

Background

Modern biological research makes possible the comprehensive study and development of heritable mutations in the mouse model at high-throughput. Using techniques spanning genetics, molecular biology, histology, and behavioral science, researchers may examine, with varying degrees of granularity, numerous phenotypic aspects of mutant mouse strains directly pertinent to human disease states. Success of these and other genome-wide endeavors relies on a well-structured bioinformatics core that brings together investigators from widely dispersed institutions and enables them to seamlessly integrate data, observations and discussions.

Description

MuTrack was developed as the bioinformatics core for a large mouse phenotype screening effort. It is a comprehensive collection of on-line computational tools and tracks thousands of mutagenized mice from birth through senescence and death. It identifies the physical location of mice during an intensive phenotype screening process at several locations throughout the state of Tennessee and collects raw and processed experimental data from each domain. MuTrack's statistical package allows researchers to access a real-time analysis of mouse pedigrees for aberrant behavior, and subsequent recirculation and retesting. The end result is the classification of potential and actual heritable mutant mouse strains that become immediately available to outside researchers who have expressed interest in the mutant phenotype.

Conclusion

MuTrack demonstrates the effectiveness of using bioinformatics techniques in data collection, integration and analysis to identify unique result sets that are beyond the capacity of a solitary laboratory. By employing the research expertise of investigators at several institutions for a broad-ranging study, the TMGC has amplified the effectiveness of any one consortium member. The bioinformatics strategy presented here lends future collaborative efforts a template for a comprehensive approach to large-scale analysis.
Versatile and open software for comparing large genomes
Authors: Kurtz Stefan , Phillippy Adam , Delcher Arthur L , Smoot Michael , Shumway Martin , Antonescu Corina , Salzberg Steven L
Published: February 2004

The newest version of MUMmer easily handles comparisons of large eukaryotic genomes at varying evolutionary distances, as demonstrated by applications to multiple genomes. Two new graphical viewing tools provide alternative ways to analyze genome alignments. The new system is the first version of MUMmer to be released as open-source software. This allows other developers to contribute to the code base and freely redistribute the code. The MUMmer sources are available at http://www.tigr.org/software/mummer.
Identification of caspase-10 in human neutrophils and its role in spontaneous apoptosis
Authors: Goepel Friederike , Weinmann Pamela , Schymeinsky Jürgen , Walzog Barbara
Published: February 2004

In the present study, we investigated the molecular mechanisms of spontaneous and tumor necrosis factor α (TNF-α)-mediated apoptosis of human polymorphonuclear neutrophils (PMN). Whereas TNF-α-mediated apoptosis was almost absent in the presence of the caspase-8 inhibitor Z-Ac-Ala-Glu-Val-Asp-7-fluoromethyl ketone (Z-AEVD-FMK), the inhibitor had no effect on spontaneous apoptosis, sggesting that spontaneous apoptosis was independent of caspase-8. Subsequently, we identified different isoforms of caspase-10 in human PMN and found high expression of caspase-10/b and/or -10/d and low expression of caspase-10/a and -10/c at the mRNA level. At the protein level, freshly isolated PMN showed high expression of caspase-10/b and -10/d as well as moderate expression of caspase-10/a and -10/c. Upon spontaneous apoptosis, caspase-10/b was down-regulated, which was accompanied by the appearance of a specific 47-kDa caspase-10/b cleavage product and an increased caspase-10 activity. In contrast, no down-regulation of caspase-10/a, -10/c, or -10/d was observed, suggesting that spontaneous apoptosis was associated with a differential activation of caspase-10/b. This was confirmed by the finding that spontaneous apoptosis was inhibited in the presence of Z-Ile-Glu-Thr-Asp (Z-IETD)-FMK, which blocks caspase-10. However, no down-regulation of caspase-10 isoforms was observed in the presence of TNF-α, suggesting that caspase-10 was not involved in TNF-α-induced apoptosis. Taken together, our study demonstrates that spontaneous and TNF-α-mediated apoptosis of PMN have different molecular requirements. Whereas TNF-α-mediated apoptosis depends on the activation of caspase-8, spontaneous apoptosis requires the activation of caspase-10/b. This finding may reveal that PMN apoptosis in different (patho-) physiological settings results from distinct molecular mechanisms.