bluuebunny/crossref_metadata_2025 huggingface dataset:
Phenotypic variation in eight extended CDKN2A germline mutation familial atypical multiple mole melanoma–pancreatic carcinoma–prone families
Authors: Lynch Henry T. , Brand Randall E. , Hogg David , Deters Carolyn A. , Fusaro Ramon M. , Lynch Jane F. , Liu Ling , Knezetic Joseph , Lassam Norman J. , Goggins Michael , Kern Scott
Published: August 2002

BACKGROUND

Hereditary pancreatic carcinoma shows extant phenotypic and genotypic heterogeneity as evidenced by its integral association with a variety of hereditary cancer syndromes inclusive of the familial atypical multiple mole melanoma (FAMMM) syndrome in concert with CDKN2A (p16) germline mutations.

METHODS

Creighton University's familial pancreatic carcinoma resource comprises 159 families of which 19 (12%) show the FAMMM cutaneous phenotypes. The authors describe eight families with the FAMMM–pancreatic carcinoma (FAMMM‐PC) association in concert with a CDKN2A germline mutation. Each family was thoroughly educated about all facets of the study, including the molecular genetics, reduced penetrance of CDKN2A mutations, and their variable expressivity. Genetic counseling was provided to each patient.

RESULTS

Diversity in cancer presentation within and among the families was noteworthy, wherein melanoma predominated in certain of the families whereas pancreatic carcinoma predominated in others. Early‐onset pancreatic carcinoma (at ages 35, 45, 46, and 49 years) appeared in some of the families whereas markedly later‐onset pancreatic carcinoma occurred in others. There were four incidences of melanoma and pancreatic carcinoma as double primaries in the same individuals. One patient with melanoma and pancreatic carcinoma had a third primary of breast carcinoma. Another patient had sarcoma, esophageal carcinoma, and two melanoma primaries, whereas his daughter had sarcoma and was a carrier of a CDKN2A mutation.

CONCLUSIONS

The authors suggest that these tumors may collectively, in concert with CDKN2A mutations, constitute a “new” putative hereditary carcinoma syndrome referred to as FAMMM‐PC. More clinical and molecular genetic research on additional families with pancreatic carcinoma in concert with the FAMMM will be required. Cancer 2002;94:84–96. © 2002 American Cancer Society.
Neuropeptides bombesin and calcitonin inhibit apoptosis‐related elemental changes in prostate carcinoma cell lines
Authors: Salido Mercedes , Vilches Jose , López Antonio , Roomans Godfried M.
Published: August 2002

BACKGROUND

Etoposide‐induced apoptosis in prostate carcinoma cells is associated with changes in the elemental content of the cells. The authors previously reported that calcitonin and bombesin inhibited etoposide‐induced apoptosis in these cells. In the current study, the authors investigated whether these neuropeptides block the etoposide‐induced changes in elemental content.

METHODS

Cells from the PC‐3 and Du 145 prostate carcinoma cell lines were grown either on solid substrates or on thin plastic films on titanium electron microscopy grids, and they were exposed to etoposide for 48 hours in the absence or presence of calcitonin and bombesin. After the exposure, the cells were frozen and freeze dried, and their elemental content was analyzed by energy‐dispersive X‐ray microanalysis in both in the scanning electron microscope and the scanning transmission electron microscope.

RESULTS

Etoposide treatment consistently induced an increase in the cellular Na concentration and a decrease in the cellular K concentration, resulting in a marked increase of the Na/K ratio and also an increase in the phosphorus:sulphur (P/S) ratio. Both bombesin and calcitonin inhibited the etoposide‐induced changes in the cellular Na/K ratio, and calcitonin, but not bombesin, inhibited the changes in the P/S ratio. No significant elemental changes were found with bombesin or calcitonin alone.

CONCLUSIONS

The neuropeptides bombesin and calcitonin, which inhibited etoposide‐induced apoptosis, also inhibited the etoposide‐induced elemental changes in prostate carcinoma cells. This important fact strengthens the link between apoptosis and changes in the intracellular elemental content. This correlation provides an objective basis for the study of neuropeptide target points and may be helpful for alternative therapeutic protocols using neuropeptide inhibitors in the treatment of patients with advanced prostatic carcinoma. Cancer 2002;94:368–77. © 2002 American Cancer Society.
ErbB2 expression is correlated with increased survival of patients with osteosarcoma
Authors: Akatsuka Tomohiro , Wada Takuro , Kokai Yasuo , Kawaguchi Satoshi , Isu Kazuo , Yamashiro Katsushige , Yamashita Toshihiko , Sawada Norimasa , Yamawaki Shinya , Ishii Seiichi
Published: August 2002

BACKGROUND

Elevated ErbB2 expression and gene amplification have been shown to be associated with poor prognosis in many cancers. Recently, it has been demonstrated that overexpression of ErbB2 protein in osteosarcoma is associated with the presence of pulmonary metastasis and decreased survival. By contrast, a previous study showed that the expression of ErbB2 declines in individual osteosarcomas as they become metastatic. In the current study, the authors determined the relation between ErbB2 status and outcome in a large number of selected patients with high‐grade osteosarcoma.

METHODS

ErbB2 status was determined immunohistochemically in biopsy specimens of osteosarcoma of the extremities from 81 patients who were treated with surgery and chemotherapy. None of the patients had metastatic disease at presentation (Stage II), and all were followed‐up for at least five years. The ErbB2 status was analyzed in relation to the lengths of event‐free and overall survival.

RESULTS

Of the 81 tumors examined, 51 (61%) demonstrated high levels of ErbB2 expression. The presence of increased levels of ErbB2 in osteosarcoma was significantly associated with the increased probability of event‐free (72.2% v. 45.6% at 5 years, P = 0.03) and overall survival (79.7% v. 58.2% at 5 years, P = 0.03). Cox multivariate analysis showed that the risk of adverse events and death was increased substantially (rate ratio: 2.24 and 2.54; 95% confidence interval, 1.07–4.72 and 1.09–5.67, respectively) among patients with decreased levels of ErbB2 protein in tumor cells, as compared with patients who had increased levels of ErbB2 in tumor cells.

CONCLUSIONS

In patients with high‐grade osteosarcoma without metastatic disease at presentation and treated with surgery and chemotherapy, the presence of increased levels of ErbB2 in tumor cells is associated with a significantly increased probability of event‐free and overall survival. Further data are needed before this marker can be used in making clinical decisions. Cancer 2002;94:1397–404. © 2002 American Cancer Society.

DOI 10.1002/cncr.10360
Close association between high serum alanine aminotransferase levels and multicentric hepatocarcinogenesis in patients with hepatitis C virus‐associated cirrhosis
Authors: Tarao Kazuo , Rino Yasushi , Ohkawa Shinichi , Tamai Setsuo , Miyakawa Kaoru , Takakura Hideki , Endo Osamu , Yoshitsugu Michiyasu , Watanabe Norihito , Matsuzaki Shohei
Published: August 2002

BACKGROUND

Multicentric development of hepatocellular carcinoma (HCC) is a characteristic feature of hepatitis C virus (HCV)‐associated cirrhosis (HCV‐LC). In this study, the objective was to determine whether the persistent elevation of the serum alanine aminotransferase (ALT) level, which represents the inflammatory necrosis of hepatocytes, is correlated with the multicentric development of hepatocellular carcinoma (HCC) in patients with early‐stage HCV‐LC.

METHODS

Ninety‐three consecutive patients with biopsy proven HCV‐LC (Child Stage A) who had been followed for > 5 years for the development of HCC were studied. They were subdivided into three groups according to their serum ALT level: Group A included 33 patients with annual average serum ALT levels that were persistently high (≥ 80 IU; high ALT group), Group B included 41 patients with annual average serum ALT levels that were persistently low (< 80 IU; low ALT group), and Group C included 19 unclassified patients. The patients had been studied prospectively with frequent ultrasonography and magnetic resonance imaging or computed tomography (CT) scans for > 5years. When the development of HCC was suspected, angiography, infusion of lipiodol into the hepatic artery, and lipiodol‐CT scans were performed in all patients to determine the number of HCC nodules.

RESULTS

In Group A, 27 patients (81.8%) developed HCC. Seventeen of 27 patients (63.0%) had multiple nodules. In contrast, in Group B, only 12 patients (29.3%) developed HCC, and only 1 of these 12 patients (8.3%) had multiple nodules. There was a significant difference between Groups A and B in the incidence of developing HCC (P < 0.001) and developing multiple nodules (P = 0.006). In addition, among the male patients, the incidence of developing multiple HCC nodules in Group A (12 of 19 patients; 63.2%) was significantly higher (P < 0.05) compared with the incidence in Group B (0 of 6 patients; 0%). The same tendency was observed among the female patients.

CONCLUSIONS

These results showed a close correlation between multicentric hepatocarcinogenesis and sustained necroinflammation of the liver in patients with HCV‐LC. Cancer 2002;94:1787–95. © 2002 American Cancer Society.

DOI 10.1002/cncr.10391
Increased proliferative activity caused by loss of p21WAF1/CIP1 expression and its clinical significance in patients with early‐stage gastric carcinoma
Authors: Noda Hiroshi , Maehara Yoshihiko , Irie Koji , Kakeji Yoshihiro , Yonemura Tomohiro , Sugimachi Keizo
Published: August 2002

BACKGROUND

Recurrences of gastric carcinoma are likely to take on a variety forms, even after patients undergo curative resection for early‐stage gastric carcinoma. It is important to identify the biologic markers that predict tumor progression and survival in these patients. Proliferating cell nuclear antigen (PCNA) acts as a processivity factor for DNA polymerase δ, which is involved directly in DNA synthesis, and the PCNA level is correlated with the proliferative state of cells. p21WAF1/CIP1 interacts with PCNA to inhibit DNA synthesis and plays a central role in regulating the cell cycle. The authors investigated patients with early‐stage gastric carcinoma to determine the clinical significance of proliferative activity and p21 expression.

METHODS

Tissue specimens from 133 Japanese patients with early‐stage gastric carcinoma that invaded the submucosal layer were immunostained with a monoclonal antibody against PCNA and p21WAF1/CIP1, and the correlations between the PCNA labeling index and p21WAF1/CIP1 expression as well as clinicopathologic factors were investigated.

RESULTS

The PCNA labeling index varied from 9.9% to 81.4%, (mean, 31.2%). The incidence of p21 positive expression was 87 of 133 patients (65.4%). The patients with a high labeling index had a significantly higher rate of lymph node metastasis (P < 0.01) and loss of p21WAF1/CIP1 expression (P < 0.05) compared with the patients with a low labeling index. The 5‐year survival rate for patients in the high labeling index group (87.0%) was significantly lower compared with the 5‐year survival rate for the patients in the low labeling index group (98.6%; P < 0.05).

CONCLUSIONS

Loss of p21WAF1/CIP1 expression contributes to the amplification of proliferative activity in patients with early‐stage gastric carcinoma. Estimation of the proliferative activity of early‐stage gastric carcinoma provides information on lymph node metastasis and prognosis. Even after patients undergo curative resection, those with early‐stage gastric carcinoma should be followed closely. Cancer 2002;94:2107–12. © 2002 American Cancer Society.

DOI 10.1002/cncr.10417
Multiple primary melanoma revisited
Authors: Blackwood M. Anne , Holmes Robin , Synnestvedt Marie , Young Megan , George Cicily , Yang Hannah , Elder David E. , Schuchter Lynn M. , Guerry DuPont , Ganguly Arupa
Published: August 2002

BACKGROUND

Incidence of cutaneous melanoma continues to increase in the Caucasian population worldwide. Approximately 5% of melanoma patients develop additional primary melanoma. This rate is significantly higher than the estimated lifetime risk of an individual for developing the disease (1.4%). These features suggest that a genetic predisposition may underlie multiple primary melanomas (MPMs). Prior studies had identified CDKN2A mutations in a few MPM individuals. The objectives of this study were to determine the frequency of family history of melanoma in MPM cases, to characterize other clinical features including history of other cancer, and to determine the association with functional CDKN2A mutations.

METHODS

This study used a case series design. All living patients who had been seen in the Pigmented Lesion Clinic at the University of Pennsylvania and who had more than one primary invasive malignant melanoma or an invasive primary followed by an in situ melanoma were eligible for participation.

RESULTS

Individuals with MPM frequently had a family history of melanoma, dysplastic nevi (DN), and/or another cancer including basal cell carcinoma (BCC), and squamous cell carcinoma breast cancer, and a personal history of DN, and basal cell carcinoma. Germline mutations in CDKN2A gene were identified in 8 of 96 MPM cases (8.3%, 95% confidence interval, 6.7–9.9%).

CONCLUSIONS

These data indicate that the presence of MPM is associated with a modest incidence of a family history of melanoma, DN, or BCC and a small association with CDKN2A mutations. Therefore, in addition to the MPM index case, other family members can benefit from screening and regular surveillance for melanoma, DN, and BCC. Cancer 2002;94:2248–55. © 2002 American Cancer Society.

DOI 10.1002/cncr.10454
Survival analysis with p27 expression and apoptosis appears to estimate the prognosis of patients with synovial sarcoma more accurately
Authors: Kawauchi Shigeto , Goto Yoshinari , Ihara Koichiro , Furuya Tomoko , Oga Atsunori , Tsuneyoshi Masazumi , Kawai Shinya , Sasaki Kohsuke
Published: August 2002

BACKGROUND

Although recent experimental studies have implicated p27 as a regulator of apoptosis in a number of neoplasms, to the authors' knowledge the relation between p27 expression and apoptosis has not yet been established in synovial sarcoma.

METHODS

The relation between p27, apoptosis, and clinicopathologic features were examined in 62 synovial sarcoma cases.

RESULTS

p27 expression was found to be correlated inversely with the extent of apoptosis in synovial sarcomas. On survival analysis, p27 expression was found to be a significant and independent prognostic factor. Although the extent of apoptosis was determined to be prognostically significant, it failed to retain an independent and significant value on multivariate analysis. The combination of p27 expression and apoptosis allowed for the more accurate estimation of prognosis in patients with synovial sarcoma than either parameter alone.

CONCLUSIONS

A combination of p27 expression and extent of apoptosis permitted the more accurate estimation ofy the prognosis of patients with synovial sarcoma. Cancer 2002; 94:2712–8. © 2002 American Cancer Society.

DOI 10.1002/cncr.10556
Mental state as a possible independent prognostic variable for survival in patients with advanced lung carcinoma
Authors: Nakahara Yasuharu , Mochizuki Yoshiro , Miyamoto Yoshihiro , Tanaka Akira , Kawamura Tetsuji , Sasaki Shin , Nakahara Yukiko , Katsura Yoshitaka
Published: August 2002

BACKGROUND

Although psychologic factors have been reported to influence the progression of cancer, this theory remains controversial. A prospective study of patients with advanced lung carcinoma was performed to explore the influence of the patient's mental state on survival.

METHODS

The patient's mental state was assessed with the Tokyo University Egogram. In a preliminary study, the egograms of long‐term survivors (survival > 3 years) with TNM Stage IIIB or Stage IV lung carcinoma were compared with the egograms of consecutive, newly diagnosed lung carcinoma patients (controls). Next, in a prospective study, 123 patients with nonsmall cell lung carcinoma and 56 patients with small cell lung carcinoma (Stage IIIB or Stage IV; Eastern Cooperative Oncology Group performance status of 0 or 1) completed the egogram. Based on the results of the preliminary study, the subjects in the prospective study were divided into Group A (Free Child [FC] ≥ 50th percentile and Adapted Child [AC] < 50th percentile) and Group B (FC < 50 percentile or AC ≥ 50 percentile). The survival of the two groups was compared. The Cox proportional hazards model was used to determine the joint effect of the patient's mental state and other prognostic factors.

RESULTS

In the preliminary study, the FC score of the long‐term survivors was significantly higher and the AC score was significantly lower than those of the controls. In the prospective study, the survival of Group A was significantly longer than that of Group B both in the nonsmall cell lung carcinoma and small cell lung carcinoma patients (P = 0.002 and P = 0.005, respectively, by the log‐rank test). Multivariate analysis demonstrated that after adjustment for clinical factors, being in Group A was a significant predictor of survival both in the nonsmall cell and small cell lung carcinoma patients.

CONCLUSIONS

The results of the current study demonstrate that the mental state of the patient as assessed by the egogram may have prognostic significance in patients with advanced lung carcinoma. Cancer 2002;94:3006–15. © 2002 American Cancer Society.

DOI 10.1002/cncr.10608
The clinical course of nonsmall cell lung carcinoma in survivors of Hodgkin disease
Authors: Laurie Scott A. , Kris Mark G. , Portlock Carol S. , Rosenzweig Kenneth E. , Miller Vincent A. , Krug Lee M. , Rusch Valerie W.
Published: August 2002

BACKGROUND

The objective of this study was to document the natural history of second lung carcinomas, common second tumors that arise in survivors of Hodgkin disease (HD).

METHODS

The data bases of the Memorial Sloan‐Kettering Cancer Center were searched to retrieve those patients who were listed with a diagnosis of both lung carcinoma and HD. Information was extracted regarding their HD (including age at diagnosis and treatment received) and their lung carcinoma (including smoking history, latency from HD, histology, disease stage, treatment received, treatment response, and survival).

RESULTS

Twenty‐one lung carcinomas were diagnosed in 19 patients, with a median latency of 13 years from the time of diagnosis of HD. Only five patients underwent complete resection, and four patients were alive and disease free at the last follow‐up. In contrast, the median survival of 14 patients with unresectable disease was 3 months. No major objective responses were documented after chemotherapy. Poor performance status and prior thoracic radiotherapy limited treatment in patients with advanced disease. All patients had either received radiotherapy to the chest for HD or had a history of smoking; 74% of patients had both risk factors for the development of lung carcinoma.

CONCLUSIONS

In patients with a history of HD, survival after the development of lung carcinoma is poor. Because surgical resection can lead to long‐term survival, early detection is crucial. HD survivors, especially those with a history of smoking, should undergo careful surveillance for second primary lung carcinomas and other diseases. Patients who are diagnosed with HD should abstain from smoking. Physicians should assess specifically the smoking status of all HD patients and prescribe a smoking cessation program. Cancer 2002;95:119–26. © 2002 American Cancer Society.

DOI 10.1002/cncr.10628
Differentiation of tyrosine hydroxylase‐synthesizing and/or aromatic L‐amino acid decarboxylase‐synthesizing neurons in the rat mediobasal hypothalamus: Quantitative double‐immunofluorescence study
Authors: Ershov Petr V. , Ugrumov Michael V. , Calas André , Krieger Monique , Thibault Jean
Published: August 2002

In this double‐immunofluorescence study, we first quantified the neurons of the arcuate nucleus as immunoreactive (+) for tyrosine hydroxylase (TH) and/or aromatic L‐amino acid decarboxylase (AADC) in rats at embryonic day 21 (E21), at postnatal day 9 (P9), and in adulthood by using conventional fluorescent or confocal microscopy. On E21, monoenzymatic (TH+AADC immunonegative (−) and TH−AADC+) neurons and bienzymatic (TH+AADC+) neurons accounted for 99% and 1%, respectively, of the whole neuron population expressing enzymes of dopamine synthesis. Further development was characterized by the dramatic increase in TH+AADC− dorsomedial and TH+AADC+ dorsomedial populations from E21 to P9 as well as by the increase in the TH+AADC+ dorsomedial population (in females) and a drop in the TH+AADC− ventrolateral and TH+AADC− dorsomedial (in males) populations from P9 to adulthood. In contrast to TH+AADC− (in males) and TH+AADC+ neurons, the TH−AADC+ neurons did not change in number from E21 to adulthood. Thus, in rat fetuses, the neurons synthesizing TH and/or AADC were mainly monoenzymatic, whereas during postnatal life the fraction of bienzymatic neurons increased by up to 60%. J. Comp. Neurol. 446:114–122, 2002. © 2002 Wiley‐Liss, Inc.
A‐fiber sensory input induces neuronal cell death in the dorsal horn of the adult rat spinal cord
Authors: Coggeshall Richard E. , Lekan Helena A. , White Fletcher A. , Woolf Clifford J.
Published: August 2002

Excitoxicity due to excessive synaptic glutamate release is featured in many neurological conditions in which neuronal death occurs. Whether activation of primary sensory pathways can ever produce sufficient over‐activity in secondary sensory neurons in the dorsal horn of the spinal cord to induce cell death, however, has not been determined. In this study, we asked whether activity in myelinated afferents (A fibers), which use glutamate as a transmitter, can induce cell death in the dorsal horn. Using stereological estimates of neuron numbers from electron microscopic sections, we found that stimulation of A‐fibers in an intact sciatic nerve at 10 Hz, 20 Hz, and 50 Hz in 10‐minute intervals at a stimulus strength that activates both Aβ and Aδ fibers resulted in the loss of 25% of neurons in lamina III, the major site of termination of large Aβ fibers, but not in lamina I, where Aδ fibers terminate. Furthermore, sciatic nerve lesions did not result in detectable neuron loss, but activation of A fibers in a previously sectioned sciatic nerve did cause substantial cell death not only in lamina III but also in laminae I and II. The expansion of the territory of A‐fiber afferent‐evoked cell death is likely to reflect the sprouting of the fibers into these laminae after peripheral nerve injury. The data show, therefore, that primary afferent A‐fiber activity can cause neuronal cell death in the dorsal horn with an anatomical distribution that depends on whether intact or injured fibers are activated. Stimulation‐induced cell death potentially may contribute to the development of persistent pain. J. Comp. Neurol. 435:276–282, 2001. © 2001 Wiley‐Liss, Inc.
Muscarinic m1 and m2 receptor proteins in local circuit and projection neurons of the primate striatum: Anatomical evidence for cholinergic modulation of glutamatergic prefronto‐striatal pathways
Authors: Alcantara Adriana A. , Mrzljak Ladislav , Jakab Robert L. , Levey Allan I. , Hersch Steven M. , Goldman‐Rakic Patricia S.
Published: August 2002

The cellular and subcellular localization of muscarinic receptor proteins m1 and m2 was examined in the neostriatum of macaque monkeys by using light and electron microscopic immunocytochemical techniques. Double‐labeling immunocytochemistry revealed m1 receptors in calbindin‐D28k–positive medium spiny projection neurons. Muscarinic m1 labeling was dramatically more intense in the striatal matrix compartment in juvenile monkeys but more intense in striosomes in the adult caudate, suggesting that m1 expression undergoes a developmental age‐dependent change. Ultrastructurally, m1 receptors were predominantly localized in asymmetric synapse‐forming spines, indicating that these spines receive extrastriatal excitatory afferents. The association of m1‐positive spines with lesion‐induced degenerating prefronto‐striatal axon terminals demonstrated that these afferents originate in part from the prefrontal cortex. The synaptic localization of m1 in these spines indicates a role of m1 in the modulation of excitatory neurotransmission. To a lesser extent, m1 was present in symmetric synapses, where it may also modulate inhibitory neurotransmission originating from local striatal neurons or the substantia nigra. Conversely, m2/choline acetyltransferase (ChAT) double labeling revealed that m2‐positive neurons corresponded to large aspiny cholinergic interneurons and ultrastructurally, that the majority of m2 labeled axons formed symmetric synapses. The remarkable segregation of the m1 and m2 receptor proteins to projection and local circuit neurons suggests a functional segregation of m1 and m2 mediated cholinergic actions in the striatum: m1 receptors modulate extrinsic glutamatergic and monoaminergic afferents and intrinsic GABAergic afferents onto projection neurons, whereas m2 receptors regulate acetylcholine release from axons of cholinergic interneurons. J. Comp. Neurol. 434:445–460, 2001. © 2001 Wiley‐Liss, Inc.
Connexin43 null mutation increases infarct size after stroke
Authors: Siushansian Ramin , Bechberger John F. , Cechetto David F. , Hachinski Vladimir C. , Naus Christian C.G.
Published: August 2002

Glial‐neuronal interactions have been implicated in both normal information processing and neuroprotection. One pathway of cellular interactions involves gap junctional intercellular communication (GJIC). In astrocytes, gap junctions are composed primarily of the channel protein connexin43 (Cx43) and provide a substrate for formation of a functional syncytium implicated in the spatial buffering capacity of astrocytes. To study the function of gap junctions in the brain, we used heterozygous Cx43 null mice, which exhibit reduced Cx43 expression. Western blot analysis showed a reduction in the level of Cx43 protein and GJIC in astrocytes cultured from heterozygote mice. The level of Cx43 is reduced in the adult heterozygote cerebrum to 40% of that present in the wild‐type. To assess the effect of reduced Cx43 and GJIC on neuroprotection, we examined brain infarct volume in wild‐type and heterozygote mice after focal ischemia. In our model of focal stroke, the middle cerebral artery was occluded at two points, above and below the rhinal fissure. Four days after surgery, mice were killed, the brains were sectioned and analyzed. Cx43 heterozygous null mice exhibited a significantly larger infarct volume compared with wild‐type (14.4 ± 1.4 mm3 vs. 7.7 ± 0.82 mm3, P < 0.002). These results suggest that augmentation of GJIC in astrocytes may contribute to neuroprotection after ischemic injury. J. Comp. Neurol. 440:387–394, 2001. © 2001 Wiley‐Liss, Inc.
Treatment of attention‐deficit hyperactivity disorder: the impact of comorbidity
Authors: Pelham William E. , Fabiano Gregory A.
Published: August 2002

A significant percentage of children with attention‐deficit hyperactivity disorder (ADHD) have comorbid, associated problems, such as learning disabilities, oppositional defiant disorder, conduct disorder, and internalizing disorders. However, it will be argued that comorbid diagnoses are not useful in treatment planning. In order to effectively treat the problematic behaviours associated with these comorbidities, clinicians must systematically assess for the impairment related to these comorbidities and implement treatment aimed to reduce impaired functioning and increase competencies in important functional domains. Clinicians who utilize this approach will find (1) that diagnosis matters little for treatment planning, while functional impairment matters a great deal, and (2) that the systematic application of behaviour modification principles improves functioning across diagnostic categories. A case study of a multiply comorbid child with ADHD is presented to illustrate effective treatment for ADHD and comorbid disorders. Copyright © 2001 John Wiley & Sons, Ltd.
School refusal: categorical diagnoses, functional analysis and treatment planning
Authors: King Neville J. , Heyne David , Tonge Bruce , Gullone Eleonora , Ollendick Thomas H.
Published: August 2002

School refusal is a complex and challenging problem for school personnel and mental health professionals. However, school refusal behaviour can be successfully treated provided there has been a sound assessment. We outline an hypothesis‐testing, multi‐informant assessment approach that incorporates diagnostic interviewing, functional analysis assessment, self‐report measures, parent/teacher checklists and review of school attendance records. Further, it is shown how assessment information facilitates individual treatment planning for the school‐refusing child and family. We also discuss the issue of graduated versus rapid school return arrangements. Copyright © 2001 John Wiley & Sons, Ltd.
Automated identification of diploid reference cells in cervical smears using image analysis
Authors: van der Laak Jeroen A.W.M. , Siebers Albertus G. , Cuijpers Vincent M.J.I. , Pahlplatz Martin M.M. , de Wilde Peter C.M. , Hanselaar Antonius G.J.M.
Published: August 2002

Background

Acquisition of DNA ploidy histograms by image analysis may yield important information regarding the behavior of premalignant cervical lesions. Accurate selection of nuclei for DNA measurement is an important prerequisite for obtaining reliable data. Traditionally, manual selection of nuclei of diagnostic and reference cells is performed by an experienced cytotechnologist. In the present study, a method for the fully automated identification of nuclei of diploid epithelial reference cells in Feulgen‐ restained Papanicolaou (PAP) smears is described.

Methods

The automated procedure consists of a decision tree implemented on the measurement device, containing nodes with feature threshold values and multivariate discriminant functions. Nodes were constructed to recognize debris and inflammatory cells, as well as diploid and nondiploid epithelial cells of the uterine cervix. Evaluation of the classifier was performed by comparing resulting diploid integrated optical densities with those from manually selected reference cells.

Results and Conclusion

On average, automatically acquired values deviated 2.4% from manually acquired values, indicating that the method described in this paper may be useful in cytometric practice. Cytometry 47:256–264, 2002. © 2002 Wiley‐Liss, Inc.
Evaluation of stabilized blood cell products as candidate preparations for quality assessment programs for CD4 T‐cell counting
Authors: Bergeron Michèle , Shafaie Atousa , Ding Tao , Phaneuf Sieglinde , Soucy Nadia , Mandy Francis , Bradley John , Fahey John
Published: August 2002

BACKGROUND

Exceptionally robust cell preparations are needed for quality assessment programs (QAPs) such as the International Program for Quality Assessment and Standardization for Immunological Measures (QASI) relevant to HIV/AIDS. A suitable product must withstand environmental stress related to transportation for a minimum of 6 days. The two objectives of this study are (1) to evaluate the performance of various commercial preparations with multicenter participation and (2) to evaluate the robustness of stabilized blood cell products. Methods: Phase 1: The performance of stabilized blood cell products was evaluated in a multicenter QAP utilizing various staining procedures and flow cytometers. Absolute cell enumeration was achieved using single‐platform T‐cell subset methodology. Phase 2: The robustness of stabilized blood cell products was evaluated by monitoring T‐cell subset values from samples stored at 4°C, 22°C, and 37°C for up to 10 days. Results: The largest interlaboratory variation in both absolute and relative T‐cell values was 16% in samples with CD4 levels ≥400 cells per microliter and 21% in samples with CD4 levels <400 cells per microliter. Six preparations retained their phenotypic expression for 7 days at 4°C and 22°C. However, only two preparations remained stable for 4 days at 37°C. Conclusion: Some stabilized cell preparations are more robust and therefore more suitable for quality assessment purposes. Cytometry (Clin. Cytometry) 50:86–91, 2002. © 2002 Wiley‐Liss, Inc.
Multiparameter flow cytometric approach for simultaneous evaluation of T lymphocyte‐endothelial cell interactions
Authors: Krupnick Alexander S. , Kreisel Daniel , Szeto Wilson Y. , Popma Sicco H. , Amin Kunjlata M. , Moore Jonni S. , Rosengard Bruce R.
Published: August 2002

Since vascular endothelium is now recognized as an immunologically active tissue, a better understanding of the relationship between endothelial cells and T lymphocytes is critical to the field of solid organ transplantation. Investigations of endothelial cell‐T cell interactions have been limited by methodology. We developed a flow cytometric method allowing for concurrent investigation of multiple cell populations within the same culture that can be applied to these complex interactions. Allogeneic CD8+ or CD4+ T cells labeled with 5,6‐carboxyfluorescein diacetate succinimidyl ester (CFSE) were added to a murine endothelial cell monolayer, in which endothelial proliferation was not inhibited by irradiation or addition of a cell cycle‐blocking agent. At specific time points, the coculture was analyzed by flow cytometry. T‐cell proliferation could be detected by gating on the T‐cell subset and evaluating the CFSE fluorescence peaks. By directly analyzing cellular division, we minimized erroneous interpretation of the data encountered by previous studies, which utilized 3H‐thymidine incorporation as sole measure of proliferation. Further subgating on cells that divided facilitated the study of CD8+ lymphocyte activation, differentiation, and acquisition of effector function. By gating on the endothelial cell population, phenotypic changes such as upregulation of surface MHC molecules or immune‐mediated apoptosis could be detected. In conclusion, we present a flow cytometric approach that could have important applications for clinical immunological monitoring in allogeneic or xenogeneic transplantation, and might provide the requisite information to better tailor immunotherapy to prevent chronic rejection. Cytometry (Comm. Clin. Cytometry) 46:271–280, 2001. © 2001 Wiley‐Liss, Inc.
Doublet discrimination in DNA cell‐cycle analysis
Authors: Wersto Robert P. , Chrest Francis J. , Leary James F. , Morris Christa , Stetler‐Stevenson MaryAlice , Gabrielson Edward
Published: August 2002

Differences in doublet analysis have the potential to alter DNA cell‐cycle measurements. The techniques for doublet determination are often used interchangeably without regard for the complexity in cell shapes and sizes of biological specimens.

G0/1 doublets were identified and quantitated using fluorescence height versus area and fluorescence width versus area pulse measurements, by enumerating the proportion of G2 + M cells that lack cyclin B1 immunoreactivity, and modeled in the DNA histograms by software algorithms. These techniques were tested on propidium iodide‐stained whole epithelial cells or nuclei from asynchronous cultures, or after exposure to chemotherapeutic agents that induced cell‐cycle arrest and were extended to human breast tumor specimens having DNA diploid patterns.

G0/1 doublets were easily discernible from G2 + M singlets in cells or nuclei that are generally homogenous and spherical in shape. Doublet discrimination based on pulse processing or cyclin B1 measurements was nonconcordant in some nonspherical cell types and in cells following cell cycle arrest. Significant differences in G0/1 doublet estimates were observed in breast tumor specimens (n = 50), with estimates based on pulse width twice those of pulse height and nearly five times greater than computer estimates. Differences between techniques are attributed to difficulties in the separation of the boundaries between G0/1 doublets and G2 + M singlet populations in biologically heterogeneous specimens.

To improve reproducibility and enhance standardization among laboratories performing cell cycle analysis in experimental cell systems and in human breast tumors, doublet discrimination analysis should best be accomplished by computer modeling. Shape and size heterogeneity of tumor and arrested cells using pulse‐processing can lead to errors and make interlaboratory comparison difficult. Cytometry (Comm. Clin. Cytometry) 46:296–306, 2001. Published 2001 Wiley‐Liss, Inc.
Dual actions of A2A adenosine receptor antagonists on motor dysfunction and neurodegenerative processes
Authors: Ongini Ennio , Monopoli Angela , Impagnatiello Francesco , Fredduzzi Silva , Schwarzschild Michael , Chen Jiang‐Fan
Published: August 2002

Of the four known adenosine receptors, the A2A receptor has received much attention over the last few years. The discovery of high‐affinity and selective A2A adenosine receptor antagonists, together with the development of different genetic lines of mice lacking A2A receptors, have greatly contributed to the new insights into the mechanisms whereby A2A receptors modulate central nervous system functions. Efforts made using the prototypic A2A receptor antagonists, e.g., the 8‐styrylxanthine KW 6002 and the pyrazolotriazolopyrimidine SCH 58261, have shown that these drugs are effective in different models of motor impairment mimicking the main features of Parkinson’s disease. Moreover, these drugs show neuroprotective properties in models of brain injury. Consistent with pharmacology, A2A receptor knockout mice have been found to be less sensitive to both motor impairment and neurochemical changes relevant to neurodegenerative disorders. The main effect of A2A receptor blockade or inactivation is related to selective interaction with dopamine‐mediated function in the striatum. However, there are responses which appear to be independent of dopamine receptors while the mechanisms underlying neuroprotection remain to be elucidated. Overall, there are now compounds that appear to be promising for treatment of Parkinson’s disease and related neurodegenerative disorders. The efforts currently ongoing to understand their efficacy in patients will make it possible to assess whether A2A receptor blockers are a new interesting class of antiparkinsonian agents. Drug Dev. Res. 52:379–386, 2001. © 2001 Wiley‐Liss, Inc.
Transnatal olfactory continuity in the rabbit: Behavioral evidence and short‐term consequence of its disruption
Authors: Coureaud Gérard , Schaal Benoist , Hudson Robyn , Orgeur Pierre , Coudert Pierre
Published: August 2002

This study investigates the role of prenatal odor learning on postnatal adaptive orientation responses in the newborn rabbit. Preference tests revealed that pups are equally attracted to the odors of placentae and colostrum (Experiments 1–4), suggesting that an odor continuity may exist between the fetal and neonatal environments. To test some predictions derived from this hypothesis, we manipulated the odor of the diet of pregnant‐lactating does to control the chemical niches of their perinates. Fetuses exposed in this way to the odor of cumin (C) were selectively attracted as neonates to the odor of pure C (Experiment 6). Prenatal exposure to C also was followed, to a certain extent, by enhanced attraction to C odor in the placenta or colostrum from females which had consumed it (Experiments 5 & 7). Finally, the functional implications of perinatal odor continuity were tested by disrupting it. The odor component of the feto–neonatal transitional environment revealed indeed to affect the ability of certain pups to gain colostrum and milk at the very first sucking opportunities (Experiment 8). © 2002 Wiley Periodicals, Inc. Dev Psychobiol 40: 372–390, 2002. Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/dev.10038
Regulation of cognitive activity and early communication development in young autistic, mentally retarded, and young normal children
Authors: Adrien Jean‐Louis , Rossignol‐Deletang Nathalie , Martineau Joëlle , Couturier Guylène , Barthelemy Catherine
Published: August 2002

Based on the Piagetian framework, this study examined regulation of cognitive activity and developmental communication profiles and their interrelationship in groups of autistic, mentally retarded, and normal children of comparable overall, verbal, and oculo‐manual developmental ages (from 6 to 24 months). Regulation of activity was assessed with both an object permanence test and an original behavior grid, and development of communication skills with the Guidetti‐Tourrette scales (French adaptation of the Seibert‐Hogan scales). The results showed evidence of certain types of dysregulation of cognitive activity and a general delay in communication ability in autistic children compared to the other two groups. Moreover, although the intensity of some of these disorders decreased in relation to the developmental levels of social interaction and joint attention in normal children, they were related to both high and low levels of development of social interaction only in autistic children. These findings raise the hypothesis of a relationship between a disorder of disengaging from an activity and developmental levels of social interaction noted at two transitory periods of early development (12 and 24 months) only in children with autism. Developmental and neuropsychological interpretations of this particular pattern are proposed. © 2001 John Wiley & Sons, Inc. Dev Psychobiol 39: 124–136, 2001
No evidence for SEL1L as a candidate gene for IDDM11‐conferred susceptibility
Authors: Pociot Flemming , Larsen Zenia M. , Zavattari Patrizia , Deidda Elisabetta , Nerup Jørn , Cattaneo Monica , Chiaramonte Raffaella , Comi Paola , Sabbadini Marta , Zollo Massimo , Biunno Ida , Cucca Francesco
Published: August 2002

Background

The SEL1L gene is located on human chromosome 14q24.3‐31 close to D14S67 which has been previously proposed to be a type 1 diabetes mellitus locus (IDDM11). Sel‐1 is a negative regulator of the Notch signalling pathway and SEL1L is selectively expressed in adult pancreas and islets of Langerhans. This suggests that SEL1L may be a candidate gene for IDDM11.

Methods

We have analysed two newly identified CA‐repeat polymorphisms within the genomic sequence of the SEL1L locus for association with type 1 diabetes mellitus (T1DM) in 152 Danish T1DM‐affected sib‐pair families and in 240 Sardinian families (229 simplex and 11 sib‐pair families).

Results

No evidence for association of the two SEL1L markers with T1DM was observed in either the Danish or the Sardinian families. We have also used allelic sharing methods to analyse linkage with T1DM in the IDDM11 region using the same markers and the Danish collection of affected sib‐pair families. No evidence of linkage was observed (Zmax=0.86).

Conclusion

Although several lines of evidence suggest that SEL1L might be a candidate for IDDM11‐conferred susceptibility to T1DM the present study does not support this hypothesis. Copyright © 2001 John Wiley & Sons, Ltd.
Effects of acipimox on the lipolysis rate in subcutaneous adipose tissue of obese subjects
Authors: Flechtner‐Mors M. , Jenkinson C. P. , Alt A. , Adler G. , Ditschuneit H. H.
Published: August 2002

Background

Acipimox is a hypolipidaemic agent reducing serum concentrations of triglycerides and non‐esterified fatty acids. Acipimox may reduce triglyceride synthesis by decreasing non‐esterified fatty acid availability from adipocytes, but this effect has yet to be demonstrated in vivo. Lipolysis after acipimox treatment was examined in subcutaneous adipose tissue of severely obese subjects with associated metabolic disorders.

Methods

The microdialysis technique was performed in abdominal subcutaneous adipose tissue of eight hyperinsulinaemic subjects. After oral treatment with acipimox, glycerol concentration was determined as an index of lipolysis rate. Blood flow was assessed by the ethanol escape technique. The rates of release of glycerol from human adipose tissue maximally stimulated by norepinephrine were also investigated in the presence of acipimox. Eight weight‐ and age‐matched subjects served as a control group.

Results

Under acipimox treatment, basal glycerol release decreased in subcutaneous adipose tissue, whereas no effect was observed on blood flow. In stimulated adipose tissue acipimox showed no effect.

Conclusion

In the present study basal glycerol outflow from adipose tissue was inhibited by acipimox. The anti‐lipolytic action of the agent may diminish elevated plasma concentrations of free fatty acids in subjects with severe obesity. Copyright © 2001 John Wiley & Sons, Ltd.
Eye defects in receptor protein‐tyrosine phosphatase α knock‐down zebrafish
Authors: Van Der Sar Astrid M. , Živković Danica , Den Hertog Jeroen
Published: August 2002

Receptor protein‐tyrosine phosphatase alpha (RPTPα) is highly expressed in the developing retina of different species, but little is known about its function there. Here, we report that injection of antisense morpholinos in zebrafish embryos reduced RPTPα expression to almost nondetectable levels up to 3 days postfertilization (dpf). RPTPα was detectable again from 4 dpf onward. RPTPα knock‐down resulted in smaller eyes. Examination of sections of the retina at different developmental stages demonstrated that already at 28 hours postfertilization (hpf) fewer cells were present in the retina of RPTPα‐morpholino–injected embryos. At 3 dpf, the layered organization of the retina was absent. In addition, the morphology and labeling with an axon specific antibody, acetylated tubulin, demonstrated that most cells appeared to be undifferentiated. Strikingly, at 5 dpf the lamination of the retina was partially restored, concomitant with re‐expression of RPTPα protein. Although cells in the retina were now differentiated, the layering of the retina remained disrupted and significant gaps were observed in the amacrine cell layer. Therefore, knock‐down of RPTPα protein provides evidence that RPTPα is essential for normal retinal development. © 2002 Wiley‐Liss, Inc.
The extracellular ATP receptor, cP2Y1, inhibits cartilage formation in micromass cultures of chick limb mesenchyme
Authors: Meyer Martin P. , Swann Karl , Burnstock Geoffrey , Clarke Jonathan D.W.
Published: August 2002

We have investigated the function of the G protein‐coupled receptor for extracellular ATP, chick P2Y1 (cP2Y1) during development of the chick limb. cP2Y1 is strongly expressed in undifferentiated limb mesenchyme cells but appears to be lost from cells as they differentiate, raising the possibility that the function of this receptor may be to inhibit cell differentiation. This pattern of expression was particularly striking surrounding areas of cartilage formation. We tested whether cP2Y1 was able to regulate cartilage formation by using an in‐vitro micromass model of chondrogenesis. Because limb cells in micromass culture lose expression of cP2Y1, we have used a gain‐of‐function approach to demonstrate that cP2Y1 expression can inhibit cartilage differentiation. We also demonstrate that early limb mesenchyme cells release ATP into the extracellular medium and have mechanisms to breakdown extracellular ATP. These results suggest that extracellular ATP, signaling through cP2Y1, can modulate the differentiation of limb mesenchyme cells in vitro, and the expression pattern of cP2Y1 suggests that this type of signaling could play a similar role in ovo. © 2001 Wiley‐Liss, Inc.