bluuebunny/crossref_metadata_2025 huggingface dataset:
Phase II Study of Ifosfamide+Doxorubicin in Patients With Advanced Synovial Sarcomas (E1793): A Trial of the Eastern Cooperative Oncology Group
Authors: Edmonson John H. , Ryan Louise M. , Falkson Carla I. , Hicks David G. , Blum Ronald H.
Published: November 2003

Purpose Because we had observed in the synovial sarcoma subgroup of a broad phase III advanced soft tissue sarcoma study a significantly greater objective regression rate from ifosfamide+doxorubicin (88%) than from doxorubicin alone (20%) (P = 0.02), the Eastern Cooperative Oncology Group (ECOG) decided to further assess this two drug combination in a subsequent Phase II study.

Patients Between 1994 and 1999, twelve adult patients with advanced synovial sarcomas were enrolled to receive, as their initial chemotherapy, ifosfamide 7.5 gm/m2 plus doxorubicin 60 mg/m2, given intravenously over two consecutive days every 3 weeks.

Methods Each day for 2 days doxorubicin 30 mg/m2 was infused over 5 min through a running i.v., followed by ifosfamide 3750 mg/m2 over 4 h. Continuous i.v. fluid was infused at 300 mL/h for 3 h on day 1, before chemotherapy was begun; then the infusion was continued at 100 mL/h for a total of 3 days. Mesna 750 mg/m2 was given 15 min before ifosfamide and at 4 and 8 h after ifosfamide on days 1 and 2 of each treatment cycle. Filgrastim (G‐CSF) 5 μg/kg was given subcutaneously each day for 14 days beginning on day 3 of each treatment cycle to limit the severity of neutropenia.

Results Five of our 12 patients (42%) experienced partial regression of their advanced synovial sarcomas; however, this first stage result was borderline for proceeding to the second planned stage of accrual and our case accrual was quite poor. Thus, the study was closed after stage one accrual. Our patients received a median of four cycles of chemotherapy (range: 1 to 6). All patients experienced at least grade 3 neutropenia (grade 4 in nine of them), and one patient died of treatment‐related sepsis following the initial cycle of chemotherapy. Median survival was 11 months.
Bisphosphonate Treatment of Benign Multifocal and Unifocal Osteolytic Tumours of Bone
Authors: Gibbons C. L. M. H. , Petra M. , Smith R. , Athanasou N. A.
Published: November 2003

Growth of benign tumours and tumour‐like lesions of bone results in osteolysis which may cause pathological fracture.
Bisphosphonates are anti‐osteolytic agents which have proved effective in the treatment of number of osteolytic conditions.
In this study we report the results of treatment with the aminobisphosphonate, pamidronate, of three benign osteolytic
tumours of bone, two cases of fibrous dysplasia and one of Langerhans cell histiocytosis. In all three cases there was clinical
and radiological improvement following treatment. Radiologically, bone lesions did not exhibit progressive enlargement.
Two cases of fibrous dysplasia also showed features suggestive of increased bone formation. These findings indicate that
bisphosphonates are likely to be useful in controlling the osteolysis of benign tumours/tumour‐like lesions of bone,
particularly in those cases where surgical intervention is not possible or multifocal lesions are present.
Hand‐Modelled Composite Prostheses after Resection of Peri‐Acetabular Bone Malignancies
Authors: Delepine G. , Delepine F. , Sokolov T. , Delepine N.
Published: November 2003

Purpose: To improve function after pelvic resection involving the acetabulum, using an anatomic composite implant built with screws and cement.

Material and method: Since 1990, 66 patients with peri‐acetabular bone malignancies have been treated by extensive resection followed by hand‐modelled innominate prosthesis with partially constrained total hip prosthesis.
The hand‐modelled innominate prosthesis was made of a titanium cup, a set of long titanium screws and two or three packs of
gentamycine‐loaded cement.

Results: Many postoperative complications were observed: deep infection (14%), hip prosthesis dislocation (25%) and local recurrence (15%).
Sixteen patients (25%) had to be reoperated. Nevertheless, at last follow‐up, 62 patients still
had composite prosthesis. The mean functional result, rated according to a modified Enneking′s staging system, was 80% with unlimited walking without support, average hip flexion 100°, length discrepancy less than 1 cm.

Discussion: These results were similar to those described in the literature for custom‐made innominate prostheses and much
better than those of alternative reconstructive procedures. Hand‐modelled composite prostheses are cheaper, easier, more
adaptable and enables better anchorage than custom‐made prostheses. Such a procedure can be used even after total iliac
wing resection.

Conclusion: The advantages of such a procedure plead for its extensive use after acetabular resection. But long‐term follow‐up is necessary to validate indications.
A Case of Liposarcoma With Peritonitis Due to Jejunal Perforation
Authors: Horiguchi Hisashi , Matsui Miwa , Yamamoto Tatsuo , Mochizuki Ryosuke , Uematsu Takeshi , Fujiwara Masachika , Ohse Hirotaka
Published: November 2003

A 21‐year‐old man, who had been treated for congenital dilatation of the bile duct 13 years previously, presented with an
acute abdomen. The physical examination suggested peritonitis, and an emergent laparotomy was performed. A perforation
was foundin the jejunum approximately 100 cm distal to the ligament of Treitz, followed by resection of a 60‐cm jejunal
segment. No tumorous lesions were found during the operation, and the resected jejunal segment showed only focal
myxomatous thickening of the serosa. Despite intensive therapy, he died of uncontrollable septic shock 2 days after the
operation. Unexpectedly, however, histological examination revealed a liposarcoma, showing an unclassifiable histology.
From the distribution of the lesion and the histological findings, it is thought that a primary lesion was somewhere else,
covered by severe adhesions due to the previous operation, and that the tumor cells spreading from it could have caused the
jejunal perforation through vascular involvement. Although extremely rare, liposarcomas in the abdomen can cause intestinal
perforation. It is important for both clinicians andpathologists to carefully investigate the cause of an unusual clinical
presentation such as intestinal perforation.
A Phase II Study of Docetaxel in Patients with Relapsed and Refractory Ewing′s Tumours
Authors: Meyer T. , Mctiernan A. , Whelan J.
Published: November 2003

Purpose. The prognosis for patients with Ewing′s tumours who have metastases at presentation or who are refractory to standard chemotherapy regimens remains poor. There is therefore a need to evaluate the role of new agents. This report describes the initial results of a prospective phase II trial of docetaxel in patients with progressive or refractory Ewing′s tumours.

Patients and methods. Fourteen patients with Ewing′s tumours who had all relapsed or progressed after treatment with multi‐drug cytotoxic therapy were treated with docetaxel 100 mg/m2 infused over 1 h, three weekly for a maximum of six cycles. Nine patients received granulocyte colony‐stimulating factor with all cycles.

Results. A partial response was observed in one patient and stable disease in two. The remaining patients progressed on treatment. The major toxicity was myelosuppression and infection with 36% patients experiencing grade 3 or 4 neutropenia and/or infection.

Conclusion. Docetaxel appears to have some activity in Ewing′s tumours even in heavily pre‐treated patients. Further evaluation of its efficacy at an earlier stage of the disease is warranted.
Delayed auditory feedback in the treatment of stuttering: clients as consumers
Authors: Van Borsel John , Reunes Gert , Van den Bergh Nathalie
Published: November 2003

Purpose: To investigate the effect of repeated exposure to delayed auditory feedback (DAF) during a 3‐month period outside a clinical environment and with only minimal clinical guidance on speech fluency in people who stutter. Method: A pretest‐post‐test design was used with repeated exposure to DAF during 3 months as the independent variable. Nine stuttering individuals aged between 18 and 45 years served as subjects. Videotaped samples from different speech tasks were collected before and after 3 months' exposure to DAF, each time under two conditions: first during non‐altered feedback (NAF) and subsequently during DAF.

Results: Before the repeated exposure to DAF, the percentage of stuttered words was significantly higher during NAF than during DAF. After 3 months‘ exposure to DAF, the percentage of stuttered words during NAF had dropped to a non‐significant level in comparison with the DAF condition. Comparison of the percentage of stuttered words during NAF before and after repeated exposure to DAF showed significantly less stuttering after the repeated exposure to DAF. The percentage of stuttered words during DAF was generally somewhat higher, but not significantly so, after the 3 months’ repeated exposure to DAF than before the exposure to DAF.

Conclusions: Results confirm that DAF is an effective means of reducing stuttering even when employed as the only treatment approach outside a therapeutic environment. They also suggest that DAF continues to promote fluency when used over a longer period, but also that length of exposure and fluency enhancement are probably not in a linear relationship.
Training volunteers as conversation partners for people with aphasia
Authors: Rayner Helen , Marshall Jane
Published: November 2003

Background: One of the most disabling consequences of aphasia is the way it excludes the person from conversation. A number of studies have attempted to tackle this problem by training the conversational partners of aphasic people. This study offers an evaluation of this approach.

Aims: Six volunteers were trained in conversing with people with moderate or severe aphasia. The study aimed to evaluate whether training changed the volunteers' knowledge about aphasia and their interactions with aphasic people. It also explored whether changes in the volunteers were matched by improved participation of the aphasic people in conversation.

Methods & Procedures: Volunteers were recruited from an aphasia group in Milton Keynes. They were trained as a group over three sessions (9 hours in total). The training drew on the techniques of Kagan (1998, 1999). It included presentation of information using different media, group discussions, viewing of videos and role play. The course aimed to improve (1) the volunteers‘ understanding of the nature of aphasia, (2) the volunteers' knowledge about communication strategies to use with aphasic people and (3) the skills of the volunteers in supporting aphasic people in conversation. An additional aim was to increase the aphasic subjects' participation in conversation. Two evaluation methods were employed. Specially designed questionnaires were administered before and after training to evaluate the volunteers’ knowledge and understanding. Volunteers were also videotaped in conversation with aphasic people before and after training. The videos were rated by speech and language therapists, using nine‐point rating scales, derived from Kagan (1999).

Outcomes & Results: Significant improvements were seen on the questionnaire scores and in ratings of the volunteers' videos. The rating gains were attributed to the training course, since baseline scores were stable and improvements only occurred after the training period. There were comparable gains in the participation of the aphasic subjects, which again occurred after training.

Conclusions: The study demonstrates that a short training course can change the knowledge and practice of experienced volunteers. The findings have implications for teaching generic skills to volunteers working with aphasic people.
Relationship between gestures and words in children with Down's syndrome and typically developing children in the early stages of communicative development
Authors: Iverson Jana M. , Longobardi Emiddia , Caselli M. Cristina
Published: November 2003

Background: Previous research has emphasized the importance of gesture in early communicative development. These studies have reported that gestures are used frequently during the first two years of life and may play a transitional role in the language acquisition process. Although there are now numerous descriptions of the relationship between gesture and the developing language system in typically‐developing (TD) children, relatively little is known about the nature and early development of the gesture‐language system in children with developmental disorders involving specific profiles of language delay and/or impairment.

Primary objective: The aim of this study is to compare early word and gesture use in children with DS and in typically‐developing children to investigate potential differences in the relationship between gestural and verbal communication in early language development.

Methods and Procedures: Ten children from upper‐middle class families participated in the study. The five children with DS (3 boys and 2 girls) had an average chronological age of 47.6 months, an average mental age of 22.4 months, and an average language age of 18 months. Each child with DS was matched to a typically developing child on the basis of gender, language age, and observed expressive vocabulary size. Children were videotaped for 30 minutes as they interacted spontaneously with their mothers. All communicative and intelligible gestures and words produced by the children were transcribed from the videotapes. Data analyses focused on: a) overall production of gestures and words (i.e., gesture and word tokens); b) the size of children's gestural and verbal repertoires (i.e., gesture and word types); and c) production and informational content of gesture‐word combinations.

Main outcomes and results: Although children with DS had significantly smaller gestural repertoires than their language age‐matched peers, there was no reliable difference between the two groups in the overall use of gesture. In addition, with DS produced two‐element combinations (primarily gesture‐word combinations) and did so at a rate comparable to that observed among their TD counterparts. However, no two‐word combinations were observed among children with DS, and there were also group differences in the information contained in children's gesture‐word combinations.

Conclusions: Taken together, these findings suggest that in addition to the well‐documented global delays in early communicative development, children with DS may exhibit additional pockets of delay, specifically in making the transition from one‐ to two‐word speech. Results are further discussed in terms of their implications for understanding the organization of the developing gesture‐language system and for the assessment of gesture in young children with communicative delays and disorders.
Effect of Aldose Reductase Inhibition on Interleukin‐1β‐Induced Nitric Oxide (NO) Synthesis in Vascular Tissue
Authors: Morales Juan , Dunbar Joseph C. , Ram Jeffrey L.
Published: November 2003

Glucose metabolism via sorbitol pathway
has been implicated as a possible contributor to
the diabetes‐related vascular changes. Nitric
oxide plays a major regulatory role in the vascular
dilatatory and constricted response. Also
it has been observed that diabetes causes vascular
changes leading to a decrease in nitric oxide
production. Additionally the accumulation of
sorbitol is also related to decreased nitric oxide
production. In the present study we investigated
the effect of normal and high glucose in the
presence or absence of both interleukin‐1β or
an aldose reductase inhibitor on nitric oxide
production in rat aortic rings in vitro. Aortic
rings from normal male Wistar rats were dissected
and incubated for 24 to 48 hrs in the presence of glucose (5.0 mM or 20 mM) or
with or without interleukin (20 ng/ml). Other
rings were incubated in the above media with
the addition of the aldose reductase inhibitor
(WAY 121509). Interleukin‐1β stimulated the
24 hr nitric oxide production and WAY 121509
decreased it under both low and high glucose
culture conditions. The interleukin‐1β stimulation
was continued for 72 hrs. Nitric oxide production
in response to interleukin‐1β was
greater at all time points when compared to the
incubation in media without interleukin‐1β. In
media containing WAY 121509 the nitric oxide
production was decreased. Interleukin‐1β stimulated
a greater increase in nitric oxide production
from aortic rings when incubated in high glucose when compared to normal glucose. The
inhibitory effect of aldose reductase inhibition
was reversible after 24 hr inhibition under both
normal and high glucose conditions. We conclude
that high glucose enhances the interleukin‐
1β‐induced nitric oxide synthesis and
the cytokine‐induced nitric oxide production
was inhibited by aldose reductase inhibition.
Nitric oxide production may be linked to redox
influences caused by the polyol pathway.
Streptozotocin‐Induced Diabetes Decreases Mammary Gland Lipoprotein Lipase Activity and Messenger Ribonucleic Acid in Pregnant and Nonpregnant Rats
Authors: Blanco-Dolado Laura , Martín-Hidalgo Antonia , Herrera Emilio
Published: November 2003

Diabetes mellitus is associated with a reduction
of lipoprotein lipase (LPL) activity in adipose
tissue and development of hypertriglyceridemia.
To determine how a condition of
severe insulin deficiency affects mammary
gland LPL activity and mRNA expression during
late pregnancy, streptozotocin (STZ) treated
(40 mg/kg) and non‐treated (control) virgin
and 20 day pregnant rats were studied. In control
rats, both LPL activity and mRNA were
higher in pregnant than in virgin rats. When
compared to control rats, STZ‐treated rats,
either pregnant or virgin, showed decreased
LPL activity and mRNA content. Furthermore,
mammary gland LPL activity was linearly correlated
with mRNA content, and either variable
was linearly correlated with plasma insulin levels. Thus, insulin deficiency impairs the
expression of LPL in mammary glands, revealing
the role of insulin as a modulator of the
enzyme at the mRNA expression level.
Effect of Treating Streptozotocin‐Induced Diabetic Rats With Sorbinil, Myo‐Inositol or Aminoguanidine on Endoneurial Blood Flow, Motor Nerve Conduction Velocity and Vascular Function of Epineurial Arterioles of the Sciatic Nerve
Authors: Coppey Lawrence J. , Gellett Jill S. , Davidson Eric P. , Dunlap Joyce A. , Yorek Mark A.
Published: November 2003

Previously we have demonstrated that diabetes causes impairment in vascular function of epineurial vessels, which precedes the slowing of motor nerve conduction velocity. Treatment
of diabetic rats with aldose reductase inhibitors, aminoguanidine or myo‐inositol supplementation have been shown to improve motor nerve conduction velocity and/or decreased endoneurial blood flow. However, the effect these treatments have on vascular reactivity of epineurial vessels of the sciatic nerve is unknown. In these studies we examined the effect of treating streptozotocin‐induced rats with sorbinil, aminoguanidine or myo‐inositol on motor nerve conduction velocity, endoneurial blood flow and endothelium dependent vascular relaxation of arterioles that provide circulation to the region of the sciatic
nerve. Treating diabetic rats with sorbinil, aminoguanidine or myo‐inositol improved the reduction of endoneurial blood flow and motor nerve conduction velocity. However, only sorbinil treatment significantly improved the diabetes‐induced impairment of acetylcholinemediated vasodilation of epineurial vessels of the sciatic nerve. All three treatments were efficacious in preventing the appropriate metabolic derangements associated with either activation of the polyol pathway or increased nonenzymatic glycation. In addition, sorbinil was shown to prevent the diabetes‐induced decrease in lens glutathione level. However, other markers of oxidative stress were not vividly improved by these treatments. These studies suggest that sorbinil treatment may be more effective in preventing neural dysfunction in
diabetes than either aminoguanidine or myoinositol.
Markers for Risk of Type 1 Diabetes in Relatives of Alsacian Patients With Type 1 Diabetes
Authors: Ongagna Jean Claude , Sapin Remi , Pinget Michel , Belcourt Alain
Published: November 2003

Background: The cytotoxic T lymphocyteassociated
antigen 4 gene (CTLA‐4) encode the
T cell receptor involved in the control of T cell
proliferation and mediates T cell apoptosis.
The receptor protein is a specific T lymphocyte
surface antigen that is detected on cells only
after antigen presentation. Thus, CTLA‐4 is
directly involved in both immune and autoimmune
responses and may be involved in the
pathogenesis of multiple T cell‐mediated
autoimmune disorders. There is polymorphism
at position 49 in exon 1 of the CTLA‐4 gene,
providing an A‐G exchange. Moreover, we
assessed the CTLA‐4 49 (Thr/Ala) polymorphism
in diabetic patients and first‐degree relatives
as compared to control subjects.

Research design and methods: Three loci
(HLA‐DQB1, DQA1 and CTLA‐4) were analysed in 62 type 1 diabetic patients, 72 firstdegree
relatives and 84 nondiabetic control
subjects by means of PCR‐RFLP.

Results: A significant enrichment in DQB1
alleles encoding for an amino acid different
from Asp in position 57 (NA) and DQA1 alleles
encoding for Arg in position 52 was
observed in diabetic subjects and first‐degree
relatives as compared to controls. The genotype
and allele frequencies of these polymorphisms
in type 1 diabetic patients and firstdegree
relatives differed significantly from
those of controls (p< 0.001 and 0.05 respectively).
CTLA‐49 Ala alleles frequencies were
75.8% in type 1 diabetic patients and 68.1% in
first‐degree relatives in comparison to 35.7% in
control subjects. The Ala/Ala genotype conferred
a relative risk of 18.8 (p < 0.001).

Conclusion: The CTLA‐4 49 Ala allele confers
an increased risk of type 1 diabetes, independent
of age and HLA‐DQ genetic markers.
D‐Chiro‐Inositol – Its Functional Role in Insulin Action and its Deficit in Insulin Resistance
Authors: Larner Joseph
Published: November 2003

In this review we discuss the biological significance
of D‐chiro‐inositol, originally discovered
as a component of a putative mediator of
intracellular insulin action, where as a putative
mediator, it accelerates the dephosphorylation
of glycogen synthase and pyruvate dehydrogenase,
rate limiting enzymes of non‐oxidative
and oxidative glucose disposal.

Early studies demonstrated a linear relationship
between its decreased urinary excretion
and the degree of insulin resistance present.
When tissue contents, including muscle, of type
2 diabetic subjects were assayed, they demonstrated
a more general body deficiency.
Administration of D‐chiro‐inositol to diabetic
rats, Rhesus monkeys and now to humans
accelerated glucose disposal and sensitized
insulin action.

A defect in vivo in the epimerization of myoinositol to chiro‐inositol in insulin sensitive tissues
of the GK type 2 diabetic rat has been elucidated.
Thus, administered D‐chiro‐inositol
may act to bypass a defective normal epimerization
of myo‐inositol to D‐chiro‐inositol
associated with insulin resistance and act to at
least partially restore insulin sensitivity and glucose
disposal.
Protein Tyrosine Phosphatase Activity in Insulin‐Resistant Rodent Psammomys Obesus
Authors: Meyerovitch Joseph , Balta Yigal , Ziv Ehud , Sack Joseph , Shafrir Eleazar
Published: November 2003

Phosphotyrosine phosphatase (PTPase) activity and its regulation
by overnight food deprivation were studied in Psammomys
obesus (sand rat), a gerbil model of insulin resistance and nutritionally
induced diabetes mellitus. PTPase activity was measured
using a phosphopeptide substrate containing a sequence identical
to that of the major site of insulin receptor (IR) β‐subunit autophosphorylation.
The PTPase activity in membrane fractions was 3.5‐,
8.3‐, and 5.9‐fold lower in liver, fat, and skeletal muscle, respectively,
compared with corresponding tissues of albino rat.Western
blotting of tissue membrane fractions in Psammomys showed lower
PTPase and IR than in albino rats. The density of PTPase transmembrane
protein band was 5.5‐fold lower in liver and 12‐fold
lower in adipose tissue. Leukocyte antigen receptor (LAR) and IR
were determined by specific immunoblotting and protein bands
densitometry and were also found to be 6.3‐fold lower in the liver
and 22‐fold lower in the adipose tissue in the hepatic membrane
fractions. Liver cytosolic PTPase activity after an overnight food
deprivation in the nondiabetic Psammomys rose 3.7‐fold compared
with postprandial PTPase activity, but it did not change significantly
in diabetic fasted animals. Similar fasting‐related changes
were detected in the activity of PTPase derived from membrane
fraction. In conclusion, the above data demonstrate that despite
the insulin resistance, Psammomys is characterized by low level of
PTPase activities in membrane and cytosolic fractions in all 3 major insulin responsive tissues, as well as in liver. PTPase activity does
not rise in activity as a result of insulin resistance and nutritionally
induced diabetes.
Defective Chylomicron Synthesis as a Cause of Delayed Particle Clearance in Diabetes?
Authors: Phillips Catherine , Madigan Claire , Owens Daphne , Collins Patrick , Tomkin Gerald H.
Published: November 2003

Chylomicron metabolism is abnormal in diabetes and the chylomicron
particle may play a very important role in atherosclerosis.
The aim of this study was to examine the effect of diabetes
on the metabolism of chylomicrons in cholesterol‐fed alloxan diabetic
and nondiabetic rabbits. Five diabetic rabbits and 5 control
rabbits were given [ 14C]linoleic acid and [ 3H]cholesterol by
gavage. Lymph was collected following cannulation of the lymph
duct and radiolabelled chylomicrons were isolated by ultracentrifugation.
The chylomicrons from each animal were injected into
paired control and diabetic recipients. Lymph apolipoprotein (apo)
B48, apo B100, and apo E were measured using sodium dodecyl
sulfate–polyacrylamide gradient gel electrophoresis. Mean blood
sugar of the diabetic donors and diabetic recipients were 19.7 ± 2.3
and 17.2 ± 3.2 mmol/L. Diabetic rabbits had significantly
raised plasma triglyceride (10.8 ± 13.9 versus 0.8 ± 0.5 mmol/L,
P < 0.02). There was a large increase in apo B48 in lymph chylomicrons
in the diabetic donor animals (0.19 ± 0.10 versus 0.04 ± 0.02
mg/h, P < 0.01) and apo B100 (0.22 ± 0.15 versus 0.07 ± 0.07 mg/h,
P < 0.05) and a reduction in apo E on the lymph chylomicron
particle (0.27 ± 0.01 versus 0.62 ± 0.07 mg/mg apo B, P < 0.001).
Diabetic recipients cleared both control and diabetic chylomicron
triglyceride significantly more slowly than control recipients
(P < 0.05). Clearance of control chylomicron cholesterol was delayed
when injected into diabetic recipients compared to when these
chylomicrons were injected into control recipients (P < 0.005).
Clearance of diabetic chylomicron cholesterol was significantly
slower when injected into control animals compared to control chylomicron injected into control animals (P < 0.02). In this animal
model of atherosclerosis, we have demonstrated that diabetes
leads to the production of an increased number of lipid and apo
E–deficient chylomicron particles. Chylomicron particles from the
control animals were cleared more slowly by the diabetic recipient
(both triglyceride and cholesterol). The chylomicron particles
obtained from the diabetic animals were cleared even more slowly
when injected into the diabetic recipient. Although there was an initial
delay in clearance of chylomicron triglyceride from the diabetic
particle when injected into the control animals, the clearance over
the first 15 minutes was not significantly different when compared
to the control chylomicron injected into the control animal. On the
other hand, the cholesterol clearance was significantly delayed.
Thus, diabetes resulted in the production of an increased number of
lipid‐ and apo E–deficient chylomicron particles. These alterations
account, in part, for the delay in clearance of these particles.
Fasting Decreases the Content of D‐Chiroinositol in Human Skeletal Muscle
Authors: Shashkin Pavel N. , Huang Laura C. , Larner Joseph , Vandenhoff George E. , Katz Abram
Published: November 2003

Two classes of inositol phosphoglycans have been implicated as
second messengers of insulin, one that activates pyruvate dehydrogenase
and contains D‐chiroinositol, and one that inhibits cyclic
AMP–dependent protein kinase and contains myoinositol. We examined
the effects of a 3‐day fast on muscle contents of inositols
in healthy humans. An oral glucose tolerance test was performed
and a biopsy was obtained from the quadriceps femoris muscle
after an overnight fast and after a 72‐hour fast. The 72‐hour fast
significantly increased plasma glucose (1.5‐ to 2‐fold) and insulin
(2‐ to 4‐fold) after glucose ingestion versus the values after the
overnight fast, indicating the manifestation of peripheral insulin
resistance. The 72‐hour fast resulted in an ∼20% decrease in the
muscle content of D‐chiroinositol (P < 0.02), but no change in the
myoinositol content. These data demonstrate that fasting specifically
decreases the muscle content of D‐chiroinositol in human muscle
and this may contribute to the finding that insulin‐mediated activation
of pyruvate dehydrogenase is attenuated after short‐term
starvation.
Hydrogen Peroxide‐Induced Inhibition of Vasomotor Activity: Evaluation of Single and Combined Treatments With Vitamin A and Insulin in Streptozotocin‐Diabetic Rats
Authors: Zobali Fulya , Besler Tanju , Ari Nuray , Karasu Çimen
Published: November 2003

A positive correlation has been established
between increased oxidative stress and cardiovascular
diseases in diabetes mellitus. We evaluated
the effects of single or combined treatments
with vitamin A (retinol acetate, 30
mg/kg/day, for 12‐weeks) and insulin (8‐10
IU/rat/day for the final 6‐week) on vasomotor
activity, oxidative stress and retinol metabolism
in 12‐week streptozotocin diabetic rats. The
vasomotor activity was determined by measuring
in vitro responsiveness of aorta rings to
phenylephrine (PE) and acetylcholine (ACh) in
the absence or in the presence of hydrogen peroxide
(H2O2). Preincubation with H2O2 (10
μM) produced a significant decrease in PE (1
mM)‐induced contraction in untreated‐diabetic
but not in control rats. Single treatment with
insulin counteracted this effect of H2O2 and
also reversed the increased contractile response
of diabetic aorta to PE, while vitamin A was
found to be ineffective. H2O2 (10 μM) also
inhibited ACh (1 mM)‐stimulated endothelium‐
dependent relaxation two fold more in diabetic
than in control aorta. In the prevention of
H2O2‐induced inhibition of vascular relaxation
to ACh, vitamin A alone was markedly effective
while insulin alone was not. The combination
of vitamin A plus insulin removed the
inhibitory action of H2O2 in diabetic aorta.
Diabetic animals displayed an increased level of aorta thiobarbituric acid reactive substance
(TBARS) in association with decreased levels of
plasma retinol and retinol‐binding protein
(RBP). Single treatment with insulin, in spite of
allowing recovery of normal growth rate and
improved glucose and retinol metabolism in
diabetic rats, was unable to control TBARS
production to the same extent as vitamin A
alone. Our findings suggest that the maintenance
of ACh‐stimulated endothelium‐dependent
vasorelaxant tone in normal physiological
levels depends largely on the prevention and/or
inhibition of peroxidative stress, which is
achieved by combined treatment with vitamin
A plus insulin. The use of vitamin A together
with insulin provides a better metabolic control
and more benefits than use of insulin alone in
the reduction of diabetes‐induced vascular
complications.
Pancreatic β‐Cell Death, Regeneration and Insulin Secretion: Roles of Poly(ADP‐Ribose) Polymerase and Cyclic ADP‐Ribose
Authors: Takasawa Shin , Okamoto Hiroshi
Published: November 2003

In the early 1980s, we proposed a unifying
model for β‐cell damage (The OKAMOTO
model), in which poly(ADP‐ribose) synthetase/
polymerase (PARP) activation plays an
essential role in the consumption of NAD+,
which leads to energy depletion and necrotic
cell death. In 1984, we demonstrated that the
administration of PARP inhibitors to 90% depancreatized
rats induces islet regeneration.
From the regenerating islet‐derived cDNA
library we isolated Reg (Regenerating Gene)
and demonstrated that Reg protein induces βcell
replication via the Reg receptor and ameliorates
experimental diabetes. More recently,
we showed that the combined addition of IL‐6
and dexamethasone induces the Reg gene expression in β‐cells and that PARP inhibitors
enhance the expression. In 1993, we found that
cyclic ADP‐ribose (cADPR), a product synthesized
from NAD+, is a second messenger for
intracellular Ca2+ mobilization for insulin secretion
by glucose, and proposed a novel mechanism
of insulin secretion, the CD38‐cADPR signal
system.

Therefore, PARP inhibitors prevent β‐cell
necrosis, induce β‐cell replication and maintain
insulin secretion.

In this paper, we would like to present a perspective
view based on our studies concerning
cell death, cell regeneration, and cell function,
especially on insulin‐producing pancreatic βcells,
in the processes of which poly(ADPribose)
synthetase/polymerase (PARP) and
cyclic ADP‐ribose (cADPR) are functioning.
The Role of Reactive Oxygen Species in Diabetes‐Induced Anomalies in Embryos of Cohen Diabetic Rats
Authors: Zangen Sarah W. , Yaffe Pirhiya , Shechtman Svetlana , Zangen David H. , Ornoy Asher
Published: November 2003

The role of the antioxidant defense mechanism in diabetesinduced
anomalies was studied in the Cohen diabetes‐sensitive
(CDs) and ‐resistant (CDr) rats, a genetic model of nutritionally
induced type 2 diabetes mellitus. Embryos, 12.5‐day‐old, of CDs
and CDr rats fed regular diet (RD) or a diabetogenic high‐sucrose
diet (HSD) were monitored for growth retardation and congenital
anomalies. Activity of superoxide dismutase (SOD) and catalaselike
enzymes and levels of ascorbic acid (AA), uric acid (UA), and
dehydroascorbic acid (DHAA) were measured in embryonic homogenates.
When fed RD, CDs rats had a decreased rate of pregnancy,
and an increased embryonic resorption. CDs embryos were
smaller than CDr embryos; 46% were maldeveloped and 7% exhibited
neural tube defects (NTDs). When fed HSD, rate of pregnancy
was reduced, resorption rate was greatly increased (56%;
P < .001), 47.6% of the embryos were retrieved without heart
beats, and 27% exhibited NTD. In contrast, all the CDr embryos
were normal when fed RD or HSD. Activity of SOD and catalase
was not different in embryos of CDs and CDr rats fedRD. When fed
HSD, levels of AA were significantly reduced, the ratio DHAA/AA
was significantly increased, and SOD activity was not sufficiently
increased when compared to embryos of CDr. The reduced fertility
of the CDs rats, the growth retardation, and NTD seem to be
genetically determined. Maternal hyperglycemia seems to result in environmentally induced embryonic oxidative stress, resulting in
further embryonic damage.
Alteration of Endothelins: A Common Pathogenetic Mechanism in Chronic Diabetic Complications
Authors: Chakrabarti Subrata , Khan Zia Ali , Cukiernik Mark , Fukuda Gen , Chen Shali , Mukherjee Suranjana
Published: November 2003

Endothelin (ET) peptides perform several physiological, vascular,
and nonvascular functions and are widely distributed in a number
of tissues. They are altered in several disease processes including
diabetes. Alteration of ETs have been demonstrated in organs
of chronic diabetic complications in both experimental and clinical
studies. The majority of the effects of ET alteration in diabetes
are due to altered vascular function. Furthermore, ET antagonists
have been shown to prevent structural and functional changes induced
by diabetes in animal models. This review discusses the contribution
of ETs in the pathogenesis and the potential role of ET
antagonism in the treatment of chronic diabetic complications.
The B Cell Antigen Receptor Controls Integrin Activity through Btk and PLCγ2
Authors: Spaargaren Marcel , Beuling Esther A. , Rurup Mette L. , Meijer Helen P. , Klok Melanie D. , Middendorp Sabine , Hendriks Rudolf W. , Pals Steven T.
Published: November 2003

Integrin-mediated adhesion and B cell antigen receptor (BCR) signaling play a critical role in B cell development and function, including antigen-specific B cell differentiation. Here we show that the BCR controls integrin α4β1 (VLA-4)-mediated adhesion of B cells to vascular cell adhesion molecule-1 and fibronectin. Molecular dissection of the underlying signaling mechanism by a combined biochemical, pharmacological, and genetic approach demonstrates that this BCR-controlled integrin-mediated adhesion requires the (consecutive) activation of Lyn, Syk, phosphatidylinositol 3-kinase, Bruton's tyrosine kinase (Btk), phospholipase C (PLC)γ2, IP3R-mediated Ca2+ release, and PKC. In contrast, activation of mitogen-activated protein kinase kinase (MEK) or extracellular signal–regulated kinase (ERK) is not required, and simultaneous activation of MEK, ERK, and PKB is not sufficient either. Furthermore, Btk is also involved in the control of integrin-mediated adhesion of preB cells. The control of integrin α4β1-mediated B cell adhesion by the BCR involves cytoskeletal reorganization and integrin clustering. These results reveal a novel function for the BCR and Btk, i.e., regulation of integrin α4β1 activity, thereby providing new insights into the control of B cell development and differentiation, as well as into the pathogenesis of the immunodeficiency disease X-linked agammaglobulineamia (XLA).
LIME
Authors: Brdičková Naděžda , Brdička Tomáš , Angelisová Pavla , Horváth Ondrej , Špička Jiří , Hilgert Ivan , Pačes Jan , Simeoni Luca , Kliche Stefanie , Merten Camilla , Schraven Burkhart , Hořejší Václav
Published: November 2003

Lymphocyte membrane rafts contain molecules critical for immunoreceptor signaling. Here, we report identification of a new raft-associated adaptor protein LIME (Lck-interacting molecule) expressed predominantly in T lymphocytes. LIME becomes tyrosine phosphorylated after cross-linking of the CD4 or CD8 coreceptors. Phospho-LIME associates with the Src family kinase Lck and its negative regulator, Csk. Ectopic expression of LIME in Jurkat T cells results in an increase of Csk in lipid rafts, increased phosphorylation of Lck and higher Ca2+ response to CD3 stimulation. Thus, LIME appears to be involved in regulation of T cell activation by coreceptors.
Mutation-induced Blocker Permeability and Multiion Block of the CFTR Chloride Channel Pore
Authors: Gong Xiandi , Linsdell Paul
Published: November 2003

Chloride permeation through the cystic fibrosis transmembrane conductance regulator (CFTR) Cl− channel is blocked by a broad range of anions that bind tightly within the pore. Here we show that the divalent anion Pt(NO2)42− acts as an impermeant voltage-dependent blocker of the CFTR pore when added to the intracellular face of excised membrane patches. Block was of modest affinity (apparent Kd 556 μM), kinetically fast, and weakened by extracellular Cl− ions. A mutation in the pore region that alters anion selectivity, F337A, but not another mutation at the same site that has no effect on selectivity (F337Y), had a complex effect on channel block by intracellular Pt(NO2)42− ions. Relative to wild-type, block of F337A-CFTR was weakened at depolarized voltages but strengthened at hyperpolarized voltages. Current in the presence of Pt(NO2)42− increased at very negative voltages in F337A but not wild-type or F337Y, apparently due to relief of block by permeation of Pt(NO2)42− ions to the extracellular solution. This “punchthrough” was prevented by extracellular Cl− ions, reminiscent of a “lock-in” effect. Relief of block in F337A by Pt(NO2)42− permeation was only observed for blocker concentrations above 300 μM; as a result, block at very negative voltages showed an anomalous concentration dependence, with an increase in blocker concentration causing a significant weakening of block and an increase in Cl− current. We interpret this effect as reflecting concentration-dependent permeability of Pt(NO2)42− in F337A, an apparent manifestation of an anomalous mole fraction effect. We suggest that the F337A mutation allows intracellular Pt(NO2)42− to enter deeply into the CFTR pore where it interacts with multiple binding sites, and that simultaneous binding of multiple Pt(NO2)42− ions within the pore promotes their permeation to the extracellular solution.
Stretch of β1 Integrin Activates an Outwardly Rectifying Chloride Current via FAK and Src in Rabbit Ventricular Myocytes
Authors: Browe David M. , Baumgarten Clive M.
Published: November 2003

Osmotic swelling of cardiac myocytes and other types of cells activates an outwardly rectifying, tamoxifen-sensitive Cl− current, ICl,swell, but it is unclear whether Cl− currents also are activated by direct mechanical stretch. We tested whether specific stretch of β1-integrin activates a Cl− current in rabbit left ventricular myocytes. Paramagnetic beads (4.5-μm diameter) coated with mAb to β1-integrin were applied to the surface of myocytes and pulled upward with an electromagnet while recording whole-cell current. In solutions designed to isolate anion currents, β1-integrin stretch elicited an outwardly rectifying Cl− current with biophysical and pharmacological properties similar to those of ICl,swell. Stretch-activated Cl− current activated slowly (t1/2 = 3.5 ± 0.1 min), partially inactivated at positive voltages, reversed near ECl, and was blocked by 10 μM tamoxifen. When stretch was terminated, 64 ± 8% of the stretch-induced current reversed within 10 min. Mechanotransduction involved protein tyrosine kinase. Genistein (100 μM), a protein tyrosine kinase inhibitor previously shown to suppress ICl,swell in myocytes, inhibited stretch-activated Cl− current by 62 ± 6% during continued stretch. Because focal adhesion kinase and Src are known to be activated by cell swelling, mechanical stretch, and clustering of integrins, we tested whether these tyrosine kinases mediated the response to β1-integrin stretch. PP2 (10 μM), a selective blocker of focal adhesion kinase and Src, fully inhibited the stretch-activated Cl− current as well as part of the background Cl− current, whereas its inactive analogue PP3 (10 μM) had no significant effect. In addition to activating Cl− current, stretch of β1-integrin also appeared to activate a nonselective cation current and to suppress IK1. Integrins are the primary mechanical link between the extracellular matrix and cytoskeleton. The present results suggest that integrin stretch may contribute to mechano-electric feedback in heart, modulate electrical activity, and influence the propensity for arrhythmogenesis.
Engineering of a Water-Soluble Plant Cytochrome P450, CYP73A1, and NMR-Based Orientation of Natural and Alternate Substrates in the Active Site
Authors: Schoch Guillaume A. , Attias Roger , Belghazi Maya , Dansette Patrick M. , Werck-Reichhart Danièle
Published: November 2003

CYP73A1 catalyzes cinnamic acid hydroxylation, a reaction essential for the synthesis of lignin monomers and most phenolic compounds in higher plants. The native CYP73A1, initially isolated from Jerusalem artichoke (Helianthus tuberosus), was engineered to simplify purification from recombinant yeast and improve solublity and stability in the absence of detergent by replacing the hydrophobic N terminus with the peptitergent amphipathic sequence PD1. Optimized expression and purification procedures yielded 4 mg engineered CYP73A1 L–1 yeast culture. This water-soluble enzyme was suitable for 1H-nuclear magnetic resonance (NMR) investigation of substrate positioning in the active site. The metabolism and interaction with the enzyme of cinnamate and four analogs were compared by UV-visible and 1H-NMR analysis. It was shown that trans-3-thienylacrylic acid, trans-2-thienylacrylic acid, and 4-vinylbenzoic acid are good ligands and substrates, whereas trans-4-fluorocinnamate is a competitive inhibitor. Paramagnetic relaxation effects of CYP73A1-Fe(III) on the 1H-NMR spectra of cinnamate and analogs indicate that their average initial orientation in the active site is parallel to the heme. Initial orientation and distances of ring protons to the iron do not explain the selective hydroxylation of cinnamate in the 4-position or the formation of single products from the thienyl compounds. Position adjustments are thus likely to occur during the later steps of the catalytic cycle.
Seed 1-Cysteine Peroxiredoxin Antioxidants Are Not Involved in Dormancy, But Contribute to Inhibition of Germination during Stress
Authors: Haslekås Camilla , Viken Marte K. , Grini Paul E. , Nygaard Vigdis , Nordgard Silje H. , Meza Trine J. , Aalen Reidunn B.
Published: November 2003

Peroxiredoxins (Prx) are thiol-dependent antioxidants containing one (1-cysteine [-Cys]) or two (2-Cys) conserved Cys residues that protect lipids, enzymes, and DNA against reactive oxygen species. In plants, the 1-Cys Prxs are highly expressed during late seed development, and the expression pattern is dormancy related in mature seeds. We have expressed the Arabidopsis 1-Cys Prx AtPER1 in Escherichia coli and show that this protein has antioxidant activity in vitro and protects E. coli in vivo against the toxic oxidant cumene hydroperoxide. Although some 1-Cys Prxs are targeted to the nucleus, a green fluorescent protein-AtPER1 fusion protein was also localized to the cytoplasm in an onion epidermis subcellular localization assay. It has been proposed that seed Prxs are involved in maintenance of dormancy and/or protect the embryo and aleurone layer surviving desiccation against damage caused by reactive oxygen species. These hypotheses were tested using transgenic Arabidopsis lines overexpressing the barley (Hordeum vulgare) 1-Cys PER1 protein and lines with reduced levels of AtPER1 due to antisensing or RNA interference. We found no correlation between Prx levels and the duration of the after-ripening period required before germination. Thus, Prxs are unlikely to contribute to maintenance of dormancy. RNA interference lines almost devoid of AtPER1 protein developed and germinated normally under standard growth room conditions. However, seeds from lines overexpressing PER1 were less inclined to germinate than wild-type seeds in the presence of NaCl, mannitol, and methyl viologen, suggesting that Prx can sense harsh environmental surroundings and play a part in the inhibition of germination under unfavorable conditions.
Two Lily SEPALLATA-Like Genes Cause Different Effects on Floral Formation and Floral Transition in Arabidopsis
Authors: Tzeng Tsai-Yu , Hsiao Chih-Chi , Chi Pei-Ju , Yang Chang-Hsien
Published: November 2003

Two AGL2-like MADS-box genes, Lily MADS Box Gene (LMADS) 3 and LMADS4, with extensive homology of LMADS3 to the Arabidopsis SEPALLATA3 were characterized from the lily (Lilium longiflorum). Both LMADS3 and LMADS4 mRNA were detected in the inflorescence meristem, in floral buds of different developmental stages, and in all four whorls of the flower organ. LMADS4 mRNA is also expressed in vegetative leaf and in the inflorescence stem where LMADS3 expression is absent. Transgenic Arabidopsis, which ectopically expresses LMADS3, showed novel phenotypes by significantly reducing plant size, flowering extremely early, and loss of floral determinacy. By contrast, 35S::LMADS4 transgenic plants were morphologically indistinguishable from wild-type plants. The early-flowering phenotype in 35S::LMADS3 transgenic Arabidopsis plants was correlated with the up-regulation of flowering time genes FT, SUPPRESSOR OF OVEREXPRESSION OF CO 1, LUMINIDEPENDENS, and flower meristem identity genes LEAFY and APETALA1. This result was further supported by the ability of 35S::LMADS3 to rescue the late-flowering phenotype in gigantea-1 (gi-1), constans-3 (co-3), and luminidependens-1 but not for ft-1 or fwa-1 mutants. The activation of these flowering time genes is, however, indirect because their expression was unaffected in plants transformed with LMADS3 fused with rat glucocorticoid receptor in the presence of both dexamethasone and cycloheximide.