bluuebunny/crossref_metadata_2025 huggingface dataset:
The effects of latanoprost and brimonidine on blood flow velocity of the retrobulbar vessels: a 3‐month clinical trial
Authors: Inan Ümit Übeyt , Ermis Sitki Samet , Yücel Aylin , Öztürk Faruk
Published: May 2003

Objective:  To investigate the effects of topical latanoprost 0.005% and topical brimonidine tartrate 0.2% on retrobulbar blood flow in patients with primary open‐angle glaucoma (POAG) and ocular hypertension (OHT).

Methods:  Forty‐one consecutive patients with POAG and OHT were enrolled in this prospective, open‐label, randomized, parallel study. Patients received either latanoprost 0.005% or brimonidine 0.2% for 3 months. Baseline retrobulbar blood flow measurements of the ophthalmic artery, central retinal artery and temporal short posterior ciliary artery were taken using colour Doppler imaging ultrasound, concurrently with systemic blood pressure, heart rate, ocular perfusion pressure and intraocular pressure (IOP) measurements. These measurements were repeated after 3 months.

Results:  Both latanoprost and brimonidine significantly reduced IOP (p < 0.05). While there was a statistically significant increase in peak systolic velocity of the ophthalmic artery, no significant change was observed in the other vessels with latanoprost treatment (p < 0.05). Topical brimonidine did not significantly alter flow velocities or resistive indices in the retrobulbar vessels after 3 months.

Conclusion:  Topical latanoprost and brimonidine significantly reduced IOP in patients with POAG and OHT without causing significant haemodynamic alterations in the retrobulbar vessels.
Prenatal exposure to influenza as a risk factor for adult schizophrenia
Authors: Limosin F. , Rouillon F. , Payan C. , Cohen J‐M. , Strub N.
Published: May 2003

Objective:  Several, but not all epidemiological studies, have demonstrated a positive correlation between exposure to the virus during the second trimester of pregnancy and an increased risk to the infants for subsequently developing schizophrenia. The present study is the first be designed in France to examine the risk of gestational exposure to the influenza virus and subsequent development of schizophrenia.

Method:  A total of 974 adults with schizophrenia born between 1949 and 1981 were compared for risk of exposure to influenza with their non‐schizophrenic siblings and with matched control patients.

Results:  Significantly more schizophrenic subjects than controls (both groups) had been exposed to the influenza virus during the fifth month of pregnancy (OR=2.24, CI: 1.49–3.35, and OR=1.61, CI: 1.04–2.49).

Conclusion:  These results suggest that influenza infection during pregnancy is a neurodevelopmental risk factor for schizophrenia in adult life.
Olanzapine in the treatment‐resistant, combat‐related PTSD – a series of case reports
Authors: Jakovljević M. , Šagud M. , Mihaljević‐Peleš A.
Published: May 2003

Objective:  Nightmares and insomnia in combat‐related post‐traumatic stress disorder (PTSD) might be resistant to treatment with selective serotonin reuptake inhibitors (SSRIs) and benzodiazepines.

Method:  We describe five case reports of patients suffering from long‐lasting and intractable nightmares and insomnia. They were given different psychotropic agents in past few years, with no improvement in their sleep disturbance. Olanzapine was added to the current treatment regimen.

Results:  Both nightmares and insomnia improved rapidly after olanzapine institution in all of five patients. No adverse events of olanzapine were reported.

Conclusion:  Olanzapine augmentation might be useful in alleviating treatment‐resistant nightmares and insomnia in patients with combat‐related PTSD.
A longitudinal analysis of the impact of social and clinical characteristics on the costs of schizophrenia treatment
Authors: Kilian R. , Matschinger H. , Becker T. , Angermeyer M. C.
Published: May 2003

Objective:  The aim of the study was the longitudinal analysis of the influence of social and clinical factors on the mid‐term costs of schizophrenia treatment.

Method:  Treatment costs as well as clinical and social characteristics of 307 patients with the ICD‐10 diagnosis of schizophrenia were assessed at five follow‐ups over 2.5 years. Between and within effects of clinical and social characteristics on treatment costs were estimated by error component regression models.

Results:  Effects caused by differences between individuals were found for age, partnership, in‐patient history, objective and subjective role functioning, life‐events and psychotic relapse. Effects of idiosyncratic transitory changes of social and clinical characteristics were found for symptoms, psychotic relapse, and for social role‐functioning.

Conclusion:  Treatment costs can be reduced to a limited extent not only by the prevention of psychotic symptoms and relapse but also by the improvement of role‐functioning capacities.
Prediction of transition from cognitive impairment to senile dementia: a prospective, longitudinal study
Authors: Artero S. , Tierney M. C. , Touchon J. , Ritchie K.
Published: May 2003

Objective: The purpose of this investigation was to replicate the statistical approach used in a previous investigation (Toronto study) within a French population to determine the best predictive model for Alzheimer's disease (AD).

Method: Data from neuropsychological tests from two prospective studies were entered into a regression model.

Results: Replication of the statistical approach in the Montpellier sample produced a three‐test model with a specificity of 99% and sensitivity of 73%. This model consisted of a delayed auditory verbal recall test, a construction test, a category fluency test and provides probability estimates for the transition to dementia in individual cases.

Conclusion: The models derived from these two longitudinal studies provide an empirical basis for the selection of tests for the definition of mild cognitive impairment of the Alzheimer type (MCI‐A). The small set of tests derived are suitable for use in general practice.
The control of apoptosis in lymphocyte selection
Authors: Strasser Andreas , Bouillet Philippe
Published: May 2003

Summary:  The stochastic nature of rearrangement and diversification of the gene segments encoding immunoglobulins (Igs) and T cell receptors (TCRs) inevitably gives rise to immature B and T lymphocytes that lack antigen receptors or express useless or dangerous (self‐antigen‐specific) ones. Signaling through antigen receptors promotes survival, proliferative expansion and further differentiation of useful cells and deletion of the useless and dangerous ones. During immune responses, pathogen‐specific B and T lymphocytes, as well as cells of the innate immune system, undergo extensive proliferation and develop effector functions, such as antibody secretion, cytotoxicity or cytokine production. To prevent tissue damage by these effector molecules, activated lymphocytes are removed when an infection has been overcome. Together with other mechanisms, including developmental arrest and induction of unresponsiveness (anergy), programmed cell death (apoptosis) of autoreactive lymphocytes safeguards immunological tolerance to self and assists in the development of an effective immune system. We have been investigating the molecular mechanisms that control programmed cell death. This review describes some of our experiments using transgenic and knockout mice, which overexpress or lack apoptosis regulators, that led to discoveries on how life and death decisions are made during development and functioning of the immune system.
Apoptosis, tolerance, and regulatory T cells – old wine, new wineskins
Authors: Ferguson Thomas A. , Stuart Patrick M. , Herndon John M. , Griffith Thomas S.
Published: May 2003

Summary:  Antigen‐specific unresponsiveness (or tolerance) has always been an important area of research. Interest in the fate of apoptotic cells and their ability to tolerize has revived interest in some of the older models involving hapten‐modified self. Recently, we have examined the mechanisms by which intravenous injection of trinitrophenol‐coupled spleen cells leads to systemic tolerance. These studies have revealed an important role for Fas/Fas ligand interactions, caspases, CD40/CD40L, and regulatory CD4+ and CD8+ T cells. Extension of these studies to peripheral deletion of T‐cell antigen receptor transgenic T cells has shown that deletion and active regulation of immune responses may be important mechanisms for the control of potentially damaging autoimmune responses.
Granzyme B: a natural born killer
Authors: Lord Sarah J. , Rajotte Ray V. , Korbutt Gregory S. , Bleackley R. Chris
Published: May 2003

Summary:  A main pathway used by cytotoxic T lymphocytes (CTLs) and natural killer cells to eliminate pathogenic cells is via exocytosis of granule components in the direction of the target cell, delivering a lethal hit of cytolytic molecules. Amongst these, granzyme B and perforin have been shown to induce CTL‐mediated target cell DNA fragmentation and apoptosis. Once released from the CTL, granzyme B binds its receptor, the mannose‐6‐phosphate/insulin‐like growth factor II receptor, and is endocytosed but remains arrested in endocytic vesicles until released by perforin. Once in the cytosol, granzyme B targets caspase‐3 directly or indirectly through the mitochondria, initiating the caspase cascade to DNA fragmentation and apoptosis. Caspase activity is required for apoptosis to occur; however, in the absence of caspase activity, granzyme B can still initiate mitochondrial events via the cleavage of Bid. Recent work shows that granzyme B‐mediated release of apoptotic factors from the mitochondria is essential for the full activation of caspase‐3. Thus, granzyme B acts at multiple points to initiate the death of the offending cell. Studies of the granzyme B death receptor and internal signaling pathways may lead to critical advances in cell transplantation and cancer therapy.
Endoplasmic Reticulum Retention, Degradation, and Aggregation of Olfactory G‐Protein Coupled Receptors
Authors: Lu Min , Echeverri Fernando , Moyer Bryan D.
Published: May 2003

The mammalian olfactory G‐protein coupled receptor family is comprised of hundreds of proteins that mediate odorant binding and initiate signal transduction cascades leading to the sensation of smell. However, efforts to functionally express olfactory receptors and identify specific odorant ligand–olfactory receptor interactions have been severely impeded by poor olfactory receptor surface expression in heterologous systems. Therefore, experiments were performed to elucidate the cellular mechanism(s) responsible for inefficient olfactory receptor cell surface expression. We determined that the mouse odorant receptors mI7 and mOREG are not selected for export from the ER and therefore are not detectable at the Golgi apparatus or plasma membrane. Specifically, olfactory receptors interact with the ER chaperone calnexin, are excluded from ER export sites, do not accumulate in ER–Golgi transport intermediates at 15 °C, and contain endoglycosidase H‐sensitive oligosaccharides, consistent with olfactory receptor exclusion from post‐ER compartments. A labile pool of ER‐retained olfactory receptors are post‐translationally modified by polyubiquitination and targeted for degradation by the proteasome. In addition, olfactory receptors are sequestered into ER aggregates that are degraded by autophagy. Collectively, these data demonstrate that poor surface expression of olfactory receptors in heterologous cells is attributable to a combination of ER retention due to inefficient folding and poor coupling to ER export machinery, aggregation, and degradation via both proteasomal and autophagic pathwaysPlasmids .
Noma (cancrum oris) associated with oral myiasis in an adult
Authors: Aguiar ÁMM , Enwonwu CO , Pires FR
Published: May 2003

Noma is a devastating oro‐facial necrotic condition affecting debilitated subjects. Oral myiasis is an infectious disease caused by deposition of larval flies in oral wounds and lesions. Oro‐facial noma–myiasis association has not been previously reported in the literature. The aim of this paper is to report a case of noma associated with myiasis in a 65‐year‐old Brazilian male.
The effect of melatonin administration on pituitary hormone secretion in man
Authors: Forsling Mary L. , Wheeler M. J. , Williams A. J.
Published: May 2003

OBJECTIVE

Evidence is accumulating that the nocturnal increase in melatonin may influence pituitary hormone secretion. The aim of this study was to determine the effect of exogenous melatonin, in concencetrations spanning the physiological range, on the release of pituitary hormones in man during daylight hours

DESIGN

A double blind, randomized, crossover study.

SUBJECTS

Eight healthy male volunteers with a mean age of 21 ± 0.5 years were studied on four occasions, observations being made after the adminstration of melatonin in doses of 0.05, 0.5 or 5.0 mg or placebo. They refrained from taking heavy exercise, alcohol and from smoking for 24 h prior to the study.

MEASUREMENTS

Serum cortisol, growth hormone, prolactin and plasma oxytocin, vasopressin, sodium, osmolality and packed cell volume were measured in samples taken at 30 minutes intervals for 150 minutes after the administration of melatonin.

RESULTS

Melatonin produced dose‐dependent changes in circulating concentrations of oxytocin and vasopressin, the 0.5 mg dose being stimulatory, while 5.0 mg was inhibitory. These two doses stimulated growth hormone release, while there was no significant effect on prolactin or cortisol release.

CONCLUSIONS

These results confirm that the nocturnal increase in melatonin could contribute to the patterns of oxytocin, vasopressin and growth hormone release seen over 24 h.
Malignant human renin producing paraganglionoma–localization with 123I‐MIBG and treatment with 131I‐MIBG
Authors: Arver Stefan , Jacobsson Hans , Cedermark Björn , Hamberger Bertil , Lundell Göran , Grimelius Lars , Brismar Kerstin
Published: May 2003

A 22‐year‐old woman presenting severe hypertension and hypokalaemia is described. Initial evaluation showed a large tumour localized at the position of the left adrenal gland. Subsequent surgery temporarily relieved all signs and symptoms caused by the tumour. The symptoms relapsed after a 2‐year disease‐free interval. At re‐evaluation, the tumour was shown to produce an uncontrolled secretion of renin, thus triggering aldosterone‐dependent hypertension. This report describes the diagnosis, treatment and clinical course of this unique patient with a malignant paraganglionoma of adrenal origin.
ACTH and cortisol release following intravenous desmopressin: a dose–response study
Authors: Scott Lucinda V. , Medbak Sami , Dinan Timothy G.
Published: May 2003

OBJECTIVE

Desmopressin (DDAVP) is a synthetic analogue of AVP, the companion regulator of corticotrophin‐releasing hormone (CRH) in the control of ACTH synthesis and release from the pituitary corticotrophs. The body of evidence from human studies suggests that DDAVP alone, unlike AVP, does not bring about ACTH release, although recent evidence suggests idiosyncracies of response in healthy subjects. We examined whether DDAVP exerted any consistent effect on ACTH and cortisol release, and also if this occurred in a dose‐dependant manner.

DESIGN AND SUBJECTS

A total of 18 subjects participated in the study. Saline, 5 μg, 10 μg and 15 μg DDAVP were administered as an intravenous bolus at 1300 h; 5, 7, 18 and 8 subjects, respectively, participated in each arm of the study. Plasma ACTH and cortisol responses were measured over a 120‐minutes period.

RESULTS

Significant between group comparisons were demonstrated for both ACTH (P < 0.05) and cortisol responses (P < 0.005) measured as maximum increment from baseline. The ACTH response to 5, 10 and 15 μg DDAVP was significantly greater than saline at all three doses, whilst maximal responses were seen at 10 μg. The cortisol responses to 10 and 15 μg DDAVP doses, but not 5 μg, were significantly greater than following saline. 11/18 subjects were deemed ‘responders’ following 10μg DDAVP on the basis of both ACTH and cortisol output.

CONCLUSIONS

This data suggests that DDAVP is capable of stimulating ACTH and cortisol release when administered alone as a bolus in over 50% of healthy subjects. This is in contrast to much of the extant literature. The mode of administration may be pertinent to this effect. This finding has implications for the recent focus on DDAVP as a diagnostic tool in disorders such as Cushing's Disease.
A new system for laser‐UVA‐microirradiation of living cells
Authors: Walter J. , Cremer T. , Miyagawa K. , Tashiro S.
Published: May 2003

Local nuclear irradiation of living cells has been used to gain insight into the dynamic changes that cell nuclei undergo in response to DNA damage. In particular, the effects of DNA double‐strand breaks (DSBs), a major threat to the genomic integrity of cells, have been studied by local nuclear irradiation with ionizing radiation. This method has the disadvantage that it requires expensive equipment to generate a sufficiently high density of focused or collimated ionizing radiation. After appropriate sensitization of the cellular DNA, nuclear microirradiation with UVA can also produce DSBs. In this communication we present a semi‐automatic system for laser‐UVA‐microirradiation based on a commercial laser scanning microscope. The system allows the convenient selection and precise irradiation of living cells, and could provide the basis for a more widespread availability of microirradiation facilities for DNA‐repair research.
Reducing image distortions due to temperature‐related microscope stage drift
Authors: Adler J. , Pagakis S. N.
Published: May 2003

Distortions in confocal 3D image data sets were related to movements of the microscope stage that matched fluctuations in laboratory temperature. Movement was apparent in all three axes. Adding a draught‐proof enclosure, covering the stage, objective lenses and supporting structures, minimized these short‐term fluctuations. However, the stage still tracked slow changes in laboratory temperature and was sensitive to heat sources mounted on the microscope. Suggestions are made about microscope design.
Cardiac autonomic characteristics in infants sleeping with their head covered by bedclothes
Authors: Franco Patricia , Lipshut Willy , Valente Filomena , Adams Stephan , Groswasser José , Kahn André
Published: May 2003

The risk of Sudden Infant Death Syndrome is increased in infants sleeping with their head covered by bedding items. This study was designed to evaluate cardiac autonomic nervous controls in infants sleeping with the head covered by bedclothes. Sixteen healthy infants with a median age of 12 weeks (range 9–13 weeks) were recorded polygraphically for one night. While they slept in their usual supine position, a bedsheet was placed over their head for about 45 min. All infants were challenged with the head covered and with the head free during both rapid eye movement (REM) and non‐rapid eye movement (NREM) sleep. Sleep, breathing and heart rate (HR) characteristics were recorded simultaneously, together with rectal and pericephalic temperatures. In both head‐free and head‐covered conditions, autoregressive spectral analysis of HR was evaluated as a function of sleep stages. During the head‐covered periods, parasympathetic tonus decreased and sympathetic activity increased in both REM and NREM sleep. Compared with the head‐free periods, the head‐covered sleep periods were characterized by greater rectal (P = 0.012) and pericephalic temperatures (P = 0.002). Covering the infant's head with a bedsheet was associated with significant changes in autonomic balance. The finding could be related to an elevation in temperatures within the infant's microenvironment.
Chronic Management of Seizures in the Syndromes of Idiopathic Generalized Epilepsy
Authors: Bourgeois Blaise F. D.
Published: May 2003

Summary: As a group, idiopathic generalized epilepsies (IGEs) have the highest rates of complete seizure control with medication. However, there are little evidence‐based data to guide drug choice for treatment. Examples of IGE include absence epilepsy, generalized tonic‐clonic epilepsy, and juvenile myoclonic epilepsy. Generalized epilepsies seem to be particularly vulnerable to seizure aggravation, and medications that are primarily effective against partial seizures are more commonly involved in seizure aggravation than other medications. A review of current research has shown that only a few medications can control IGE without potentially causing seizure aggravation. Broad‐spectrum antiepileptic drugs such as valproate (VPA), lamotrigine, and topiramate are extremely effective at controlling a variety of seizures without causing excessive seizure aggravation. Among these drugs, VPA has the longest clinical experience history and the largest body of published data.
Change in the Retrograde Atrial Activation Sequence Following Radiofrequency Modification of the Atrioventricular Node:
Authors: Dizon José , Reiffel James , Kassotis John , Woollett Ian , Garan Hasan
Published: May 2003

Introduction: Despite the great success in treating AV nodal reentrant tachycardia (AVNRT) with radiofrequency modification of the AV node, the dimensions of the electrophysiologic circuit of this arrhythmia remain unclear, and simple models fail to explain all tachycardia‐related phenomena.

Methods and Results: We describe three unusual cases of supraventricular tachycardia (SVT). In all three cases, retrograde atrial activation during ventricular pacing or during SVT manifested local left atrial electrograms recorded from the coronary sinus preceding the septal atrial electrograms (eccentric activation), with earliest atrial activity at the lateral or posterolateral mitral annulus. Electrophysiologic maneuvers and observations were consistent with AVNRT as the mechanism in each case. In all cases, radiofrequency modification of the AV node eliminated inducible SVT and abolished dual pathway AV nodal physiology. The retrograde atrial activation sequence during ventricular pacing changed after ablation in each case, with septal atrial electrograms preceding left atrial electrograms recorded from the coronary sinus (concentric activation).

Conclusion: The observations in these cases cannot be explained by the traditional model of slow, fast, and intermediate AV nodal pathways. A model incorporating a circuit close to the AV node with left atrial and coronary sinus connections is proposed.
(J Cardiovasc Electrophysiol, Vol. 14, pp. 461‐466, May 2003)
Effect of Amiodarone on Dispersion of Atrial Refractoriness and Cycle Length in Patients with Atrial Fibrillation
Authors: Fynn Simon P. , Todd Derick M. , Julian W. , Hobbs C. , Armstrong Karen L. , Fitzpatrick P. , Garratt Clifford J.
Published: May 2003

Introduction: Amiodarone is effective in preventing the recurrence of atrial fibrillation (AF) after cardioversion (CV). Dispersion of atrial refractoriness may be relevant to the generation of AF. We designed a study to determine the electrophysiologic effects of amiodarone in patients with previous early recurrence of AF after CV.

Methods and Results: Fifteen patients with previous AF recurrence (without antiarrhythmic drugs) after CV (CV1) were selected for amiodarone therapy and repeat CV (CVamio). Prior to CV1, mean AF cycle length (AFCL) had been recorded at four atrial sites (right atrial appendage [RAA], distal coronary sinus [DCS], right atrial lateral wall [LAT], and interatrial septum [IAS]) and dispersion of AFCL had been calculated. These patients were treated with amiodarone and, prior to CVamio, AFCL was recorded at the four atrial sites as for CV1. Between CV1 and CVamio, AFCL increased at all atrial sites:

153 ± 13 msec

to

179 ± 14 msec

at RAA,

144 ± 12 msec

to

174 ± 18 msec

at DCS,

158 ± 13 msec

to

182 ± 16 msec

at LAT, and

161 ± 18 msec

to

181 ± 17 msec

at IAS. Dispersion of AFCL decreased from

24 ± 10 msec

at CV1 to

15 ± 11 msec

at

CVamio (P = 0.01)

. The median time in sinus rhythm increased from 3.12 hours post CV1 to 28 days post

CVamio (P < 0.02)
.

Conclusion: Amiodarone causes a reduction in the dispersion of AFCL. This action may be relevant to the beneficial effects of amiodarone in patients with AF.
(J Cardiovasc Electrophysiol, Vol. 14, pp. 485‐491, May 2003)
Characterization of Sustained Atrial Tachycardia in Dogs with Rapid Ventricular Pacing‐Induced Heart Failure
Authors: Stambler Bruce S. , Fenelon Guilherme , Shepard Richard K. , Clemo Henry F. , Guiraudon Colette M.
Published: May 2003

Introduction: Atrial arrhythmias often complicate congestive heart failure (CHF). We characterized inducible atrial tachyarrhythmias and electrophysiologic alterations in dogs with CHF and atrial enlargement produced by rapid ventricular pacing.

Methods and Results: Endocardial pacing leads were implanted in the right ventricle, right atrium, and coronary sinus in 18 dogs. The right ventricular lead was connected to an implanted pacemaker capable of rapid ventricular pacing. The atrial leads were used to perform electrophysiologic studies in conscious animals at baseline in all dogs, during CHF induced by rapid ventricular pacing at 235 beats/min in 15 dogs, and during recovery from CHF in 6 dogs. After

20 ± 7 days

of rapid ventricular pacing, inducibility of sustained atrial tachycardia (cycle length

120 ± 12 msec

) was enhanced in dogs with CHF. Atrial tachycardia required a critical decrease in atrial burst pacing cycle length (≤130 msec) for induction and often could be terminated by overdrive pacing. Calcium antagonists (verapamil, flunarizine, ryanodine) terminated atrial tachycardia and suppressed inducibility. Effective refractory periods at 400‐ and 300‐msec cycle lengths in the right atrium and coronary sinus were prolonged in dogs with CHF. Atrial cells from dogs with CHF had prolonged action potential durations and reduced resting potentials and delayed afterdepolarizations (DADs). Mitochondria from atrial tissue from dogs with CHF were enlarged and had internal cristae disorganization.

Conclusions: CHF promotes inducibility of sustained atrial tachycardia. Based on the mode of tachycardia induction, responses to pacing and calcium antagonists, and presence of DADs, atrial tachycardia in this CHF model has a mechanism most consistent with DAD‐induced triggered activity resulting from intracellular calcium overload.
(J Cardiovasc Electrophysiol, Vol. 14, pp. 499‐507, May 2003)
Successful Catheter Ablation of an Anteroseptal Accessory Pathway from the Noncoronary Sinus of Valsalva
Authors: Tada Hiroshi , Naito Shigeto , Nogami Akihiko , Taniguchi Koichi
Published: May 2003

We describe a patient who underwent radiofrequency catheter ablation of concealed left lateral and anteroseptal accessory pathways. After successful elimination of the concealed left anterolateral accessory pathway, the earliest retrograde atrial activation was located in the His‐bundle region. Complete elimination of the accessory pathway conduction was achieved with a radiofrequency energy application from the noncoronary sinus of Valsalva.
(J Cardiovasc Electrophysiol, Vol. 14, pp. 544‐546, May 2003)
Essential functions of p21-activated kinase 1 in morphogenesis and differentiation of mammary glands
Authors: Wang Rui-An , Vadlamudi Ratna K. , Bagheri-Yarmand Rozita , Beuvink Iwan , Hynes Nancy E. , Kumar Rakesh
Published: May 2003

Although growth factors have been shown to influence mammary gland development, the nature of downstream effectors remains elusive. In this study, we show that the expression of p21-activated kinase (Pak)1, a serine/threonine protein kinase, is activated in mammary glands during pregnancy and lactation. By targeting an ectopic expression of a kinase-dead Pak1 mutant under the control of ovine β-lactoglobulin promoter, we found that the mammary glands of female mice expressing kinase-dead Pak1 transgene revealed incomplete lobuloalveolar development and impaired functional differentiation. The expression of whey acidic protein and β-casein and the amount of activated Stat5 in the nuclei of epithelial cells in transgenic mice were drastically reduced. Further analysis of the underlying mechanisms revealed that Pak1 stimulated β-casein promoter activity in normal mouse mammary epithelial cells and also cooperated with Stat5a. Pak1 directly interacted with and phosphorylated Stat5a at Ser 779, and both COOH-terminal deletion containing Ser 779 of Stat5a and the Ser 779 to Ala mutation completely prevented the ability of Pak1 to stimulate β-casein promoter. Mammary glands expressing inactive Pak1 exhibited a reduction of Stat5a Ser 779 phosphorylation. These findings suggest that Pak1 is required for alveolar morphogenesis and lactation function, and thus, identify novel functions of Pak1 in the mammary gland development.
A novel human protein of the maternal centriole is required for the final stages of cytokinesis and entry into S phase
Authors: Gromley Adam , Jurczyk Agata , Sillibourne James , Halilovic Ensar , Mogensen Mette , Groisman Irina , Blomberg Maureen , Doxsey Stephen
Published: May 2003

Centrosomes nucleate microtubules and contribute to mitotic spindle organization and function. They also participate in cytokinesis and cell cycle progression in ways that are poorly understood. Here we describe a novel human protein called centriolin that localizes to the maternal centriole and functions in both cytokinesis and cell cycle progression. Centriolin silencing induces cytokinesis failure by a novel mechanism whereby cells remain interconnected by long intercellular bridges. Most cells continue to cycle, reenter mitosis, and form multicellular syncytia. Some ultimately divide or undergo apoptosis specifically during the protracted period of cytokinesis. At later times, viable cells arrest in G1/G0. The cytokinesis activity is localized to a centriolin domain that shares homology with Nud1p and Cdc11p, budding and fission yeast proteins that anchor regulatory pathways involved in progression through the late stages of mitosis. The Nud1p-like domain of centriolin binds Bub2p, another component of the budding yeast pathway. We conclude that centriolin is required for a late stage of vertebrate cytokinesis, perhaps the final cell cleavage event, and plays a role in progression into S phase.
Size-selective loosening of the blood-brain barrier in claudin-5–deficient mice
Authors: Nitta Takehiro , Hata Masaki , Gotoh Shimpei , Seo Yoshiteru , Sasaki Hiroyuki , Hashimoto Nobuo , Furuse Mikio , Tsukita Shoichiro
Published: May 2003

Tight junctions are well-developed between adjacent endothelial cells of blood vessels in the central nervous system, and play a central role in establishing the blood-brain barrier (BBB). Claudin-5 is a major cell adhesion molecule of tight junctions in brain endothelial cells. To examine its possible involvement in the BBB, claudin-5–deficient mice were generated. In the brains of these mice, the development and morphology of blood vessels were not altered, showing no bleeding or edema. However, tracer experiments and magnetic resonance imaging revealed that in these mice, the BBB against small molecules (<800 D), but not larger molecules, was selectively affected. This unexpected finding (i.e., the size-selective loosening of the BBB) not only provides new insight into the basic molecular physiology of BBB but also opens a new way to deliver potential drugs across the BBB into the central nervous system.
Oscillatory nucleocytoplasmic shuttling of the general stress response transcriptional activators Msn2 and Msn4 in Saccharomyces cerevisiae
Authors: Jacquet Michel , Renault Georges , Lallet Sylvie , De Mey Jan , Goldbeter Albert
Published: May 2003

Msn2 and Msn4 are two related transcriptional activators that mediate a general response to stress in yeast Saccharomyces cerevisiae by eliciting the expression of specific sets of genes. In response to stress or nutritional limitation, Msn2 and Msn4 migrate from the cytoplasm to the nucleus. Using GFP-tagged constructs and high-resolution time-lapse video microscopy on single cells, we show that light emitted by the microscope also triggers this migration. Unexpectedly, the population of Msn2 or Msn4 molecules shuttles repetitively into and out of the nucleus with a periodicity of a few minutes. A large heterogeneity in the oscillatory response to stress is observed between individual cells. This periodic behavior, which can be induced by various types of stress, at intermediate stress levels, is not dependent upon protein synthesis and persists when the DNA-binding domain of Msn2 is removed. The cAMP–PKA pathway controls the sensitivity of the oscillatory nucleocytoplasmic shuttling. In the absence of PKA, Msn4 continues to oscillate while Msn2 is maintained in the nucleus. We show that a computational model based on the possibility that Msn2 and Msn4 participate in autoregulatory loops controlling their subcellular localization can account for the oscillatory behavior of the two transcription factors.
Amyloid as a natural product
Authors: Kelly Jeffery W. , Balch William E.
Published: May 2003

Amyloid fibrils, such as those found in Alzheimer's and the gelsolin amyloid diseases, result from the misassembly of peptides produced by either normal or aberrant intracellular proteolytic processing. A paper in this issue by Marks and colleagues (Berson et al., 2003) demonstrates that intra-melanosome fibrils are formed through normal biological proteolytic processing of an integral membrane protein. The resulting peptide fragment assembles into fibrils promoting the formation of melanin pigment granules. These results, along with the observation that amyloid fibril formation by bacteria is highly orchestrated, suggest that fibril formation is an evolutionary conserved biological pathway used to generate natural product nanostructures.
The Effects of First Language Orthographic Features on Second Language Reading in Text
Authors: Akamatsu Nobuhiko
Published: May 2003

This study investigated the effects of first language (L1) orthographic features on second language (L2) reading. Three groups of fluent L2 readers with a variety of L1 backgrounds (i.e., Chinese, Japanese, and Persian) were provided with English passages printed in either alternated case or a normal manner and asked to read them for comprehension. Results showed that the Chinese and the Japanese (nonalphabetic L1 groups) were more adversely affected by case alternation than was the Persian group (an alphabetic L1 group), in terms of reading speed. This suggests that, because of L1 effects on basic processing in L2 reading, L2 readers with a nonalphabetic L1 background were less efficient in processing English words than those with an alphabetic L1 background.
Processing Difficulty in Comprehension and Production of Relative Clauses by Learners of English as a Second Language
Authors: Izumi Shinichi
Published: May 2003

This study tested the predictions of three major hypotheses of relative clause acquisition in second language acquisition: Keenan and Comrie's (1977) Noun Phrase Accessibility Hierarchy (NPAH), Kuno's (1974) Perceptual Difficulty Hypothesis (PDH), and Hamilton's (1994) SO Hierarchy Hypothesis (SOHH). These hypotheses are based on different rationales and make different predictions on the difficulty order of different relative clause sentence types. Analyses of the data collected from 61 learners of English as a second language in three different elicitation tasks found mixed support for the NPAH and the SOHH and full support for the PDH. Generally, the results suggest a complementary relationship between the NPAH and the PDH. Some differences observed in different tasks point to the importance of examining learners' processing problems in multiple tasks in different modalities.
Axonal Transport and Sorting of Herpes Simplex Virus Components in a Mature Mouse Visual System
Authors: LaVail Jennifer H. , Tauscher Andrew N. , Aghaian Elda , Harrabi Ons , Sidhu Sukhvinder S.
Published: May 2003

The time course for delivery and transport of two major proteins of herpes simplex virus (HSV) has been determined for mature mouse retinal ganglion cell axons in vivo. Twenty-four hours after intravitreal injection of HSV, valacyclovir was introduced into the drinking water of the mice to inhibit subsequent viral replication. Without treatment, viral spread and replication in periaxonal glial cells confound study of axonal transport. At 2 to 5 days after infection, the animals were sacrificed and contiguous segments of the optic pathway were removed. Immunofluorescence microscopy indicated that the number of infected astrocytes was reduced in the proximal optic nerve and eliminated in the optic tract. Western blots of the retina with antibodies for envelope and capsid components, glycoprotein D (gD) and VP5, respectively, revealed that both components were expressed in retinal homogenates by 2 days. Results of reverse transcription-PCR indicated that there was no gD mRNA present in the treated optic tract 5 days after infection. Therefore, we conclude that gD is transcribed from viral mRNA in the retinal ganglion cell bodies. The gD accumulated in the proximal ganglion cell axon by 2 days and reached the most distal segment after 3 days. The VP5 first appeared in the proximal axons at 4 days, about 48 h after the appearance of gD. Thus, gD entered the axon earlier and independent of VP5. These finding confirm the subassembly model of viral transport in neurons and suggest that there is a 4- to 5-day window for initiation of effective antiviral treatment with valacyclovir.
Expression and Immunogenicity of Sequence-Modified Human Immunodeficiency Virus Type 1 Subtype BpolandgagpolDNA Vaccines
Authors: zur Megede Jan , Otten Gillis R. , Doe Barbara , Liu Hong , Leung Louisa , Ulmer Jeffrey B. , Donnelly John J. , Barnett Susan W.
Published: May 2003

Control of the worldwide AIDS pandemic may require not only preventive but also therapeutic immunization strategies. To meet this challenge, the next generation of human immunodeficiency virus type 1 (HIV-1) vaccines must stimulate broad and durable cellular immune responses to multiple HIV antigens. Results of both natural history studies and virus challenge studies with macaques indicate that responses to both Gag and Pol antigens are important for the control of viremia. Previously, we reported increased Rev-independent expression and improved immunogenicity of DNA vaccines encoding sequence-modified Gag derived from the HIV-1SF2strain (J. zur Megede, M. C. Chen, B. Doe, M. Schaefer, C. E. Greer, M. Selby, G. R. Otten, and S. W. Barnett, J. Virol. 74: 2628-2635, 2000). Here we describe results of expression and immunogenicity studies conducted with novel sequence-modified HIV-1SF2GagPol and Pol vaccine antigens. These Pol antigens contain deletions in the integrase coding region and were mutated in the reverse transcriptase (RT) coding region to remove potentially deleterious enzymatic activities. The resulting Pol sequences were used alone or in combination with sequence-modified Gag. In the latter, the natural translational frameshift between the Gag and Pol coding sequences was either retained or removed. Smaller, in-frame fusion gene cassettes expressing Gag plus RT or protease plus RT also were evaluated. Expression of Gag and Pol from GagPol fusion gene cassettes appeared to be reduced when the HIV protease was active. Therefore, additional constructs were evaluated in which mutations were introduced to attenuate or inactivate the protease activity. Nevertheless, when these constructs were delivered to mice as DNA vaccines, similar levels of CD8+T-cell responses to Gag and Pol epitopes were observed regardless of the level of protease activity. Overall, the cellular immune responses against Gag induced in mice immunized with multigenicgagpolplasmids were similar to those observed in mice immunized with the plasmid encoding Gag alone. Furthermore, all of the sequence-modifiedpolandgagpolplasmids expressed high levels of Pol-specific antigens in a Rev-independent fashion and were able to induce potent Pol-specific T- and B-cell responses in mice. These results support the inclusion of agagpolin-frame fusion gene in future HIV vaccine approaches.
Human Rhinovirus Type 16: Mutant V1210A Requires Capsid-Binding Drug for Assembly of Pentamers To Form Virions during Morphogenesis
Authors: Lee Wai-Ming , Wang Wensheng
Published: May 2003

Our laboratory has previously reported isolation of human rhinovirus type 16 (HRV16) mutants which depend on WIN 52035 to grow. A rapid rise of progeny virus infectivity occurred when drug was added late in growth cycles, suggesting that the drug-dependence lesion was at the step of virus assembly (W. Wang et al., J. Virol. 72:1210-1218, 1998). Here, we report that capsid subunits, 5S protomers and 14S pentamers, of a drug-dependent mutant were produced normally in the absence of drug, but mutant 80S empty capsids and 150S provirions were not formed, maturation cleavage of provirions (VP0 → VP2 + VP4) did not occur, and the unassembled mutant capsid subunits were degraded with a half-life of 15 min. Drug was not required by mutant virus for attachment, uncoating, RNA synthesis and protein synthesis, and polyprotein processing except maturation cleavage. The requirement of drug for assembly of mutant pentamers to form provirions and the rapid assembly of preformed subunits (synthesized in the absence of drug) after drug addition suggested that after native pentamers (P5) have been formed they must be converted to an assembly active state (P5∗), possibly by a conformational change induced by the binding of drug. We propose that pocket factor plays the same role in wild-type virus. In addition, we also report the construction and the properties of a full-length cDNA clone of HRV16, pR16.11, which produces in vitro transcripts with infectivity similar to that of virion RNA. This cDNA clone is available at the American Type Culture Collection.
Binding of CCAAT Displacement Protein CDP to Adenovirus Packaging Sequences
Authors: Erturk Ece , Ostapchuk Philomena , Wells Susanne I. , Yang Jihong , Gregg Keqin , Nepveu Alain , Dudley Jaquelin P. , Hearing Patrick
Published: May 2003

Adenovirus (Ad) type 5 DNA packaging is initiated in a polar fashion from the left end of the genome. The packaging process is dependent upon the
cis
-acting packaging domain located between nucleotides 194 and 380. Seven A/T-rich repeats have been identified within this domain that direct packaging. A1, A2, A5, and A6 are the most important repeats functionally and share a bipartite sequence motif. Several lines of evidence suggest that there is a limiting
trans
-acting factor(s) that plays a role in packaging. Two cellular activities that bind to minimal packaging domains in vitro have been previously identified. These binding activities are P complex, an uncharacterized protein(s), and chicken ovalbumin upstream promoter transcription factor (COUP-TF). In this work, we report that a third cellular protein, octamer-1 protein (Oct-1), binds to minimal packaging domains. In vitro binding analyses and in vivo packaging assays were used to examine the relevance of these DNA binding activities to Ad DNA packaging. The results of these experiments reveal that COUP-TF and Oct-1 binding does not play a functional role in Ad packaging, whereas P-complex binding directly correlates with packaging function. We demonstrate that P complex contains the cellular protein CCAAT displacement protein (CDP) and that full-length CDP is found in purified virus particles. In addition to cellular factors, previous evidence indicates that viral factors play a role in the initiation of viral DNA packaging. We propose that CDP, in conjunction with one or more viral proteins, binds to the packaging sequences of Ad to initiate the encapsidation process.
Comparative Immunogenicity in Rhesus Monkeys of DNA Plasmid, Recombinant Vaccinia Virus, and Replication-Defective Adenovirus Vectors Expressing a Human Immunodeficiency Virus Type 1
gag
Gene
Authors: Casimiro Danilo R. , Chen Ling , Fu Tong-Ming , Evans Robert K. , Caulfield Michael J. , Davies Mary-Ellen , Tang Aimin , Chen Minchun , Huang Lingyi , Harris Virginia , Freed Daniel C. , Wilson Keith A. , Dubey Sheri , Zhu De-Min , Nawrocki Denise , Mach Henryk , Troutman Robert , Isopi Lynne , Williams Donna , Hurni William , Xu Zheng , Smith Jeffrey G. , Wang Su , Liu Xu , Guan Liming , Long Romnie , Trigona Wendy , Heidecker Gwendolyn J. , Perry Helen C. , Persaud Natasha , Toner Timothy J. , Su Qin , Liang Xiaoping , Youil Rima , Chastain Michael , Bett Andrew J. , Volkin David B. , Emini Emilio A. , Shiver John W.
Published: May 2003

Cellular immune responses, particularly those associated with CD3
+
CD8
+
cytotoxic T lymphocytes (CTL), play a primary role in controlling viral infection, including persistent infection with human immunodeficiency virus type 1 (HIV-1). Accordingly, recent HIV-1 vaccine research efforts have focused on establishing the optimal means of eliciting such antiviral CTL immune responses. We evaluated several DNA vaccine formulations, a modified vaccinia virus Ankara vector, and a replication-defective adenovirus serotype 5 (Ad5) vector, each expressing the same codon-optimized HIV-1
gag
gene for immunogenicity in rhesus monkeys. The DNA vaccines were formulated with and without one of two chemical adjuvants (aluminum phosphate and CRL1005). The Ad5-
gag
vector was the most effective in eliciting anti-Gag CTL. The vaccine produced both CD4
+
and CD8
+
T-cell responses, with the latter consistently being the dominant component. To determine the effect of existing antiadenovirus immunity on Ad5-
gag
-induced immune responses, monkeys were exposed to adenovirus subtype 5 that did not encode antigen prior to immunization with Ad5-
gag
. The resulting anti-Gag T-cell responses were attenuated but not abolished. Regimens that involved priming with different DNA vaccine formulations followed by boosting with the adenovirus vector were also compared. Of the formulations tested, the DNA-CRL1005 vaccine primed T-cell responses most effectively and provided the best overall immune responses after boosting with Ad5-
gag
. These results are suggestive of an immunization strategy for humans that are centered on use of the adenovirus vector and in which existing adenovirus immunity may be overcome by combined immunization with adjuvanted DNA and adenovirus vector boosting.
The Unique Vertex of Bacterial Virus PRD1 Is Connected to the Viral Internal Membrane
Authors: Strömsten Nelli J. , Bamford Dennis H. , Bamford Jaana K. H.
Published: May 2003

Icosahedral double-stranded DNA (dsDNA) bacterial viruses are known to package their genomes into preformed procapsids via a unique portal vertex. Bacteriophage PRD1 differs from the more commonly known icosahedral dsDNA phages in that it contains an internal lipid membrane. The packaging of PRD1 is known to proceed via preformed empty capsids. Now, a unique vertex has been shown to exist in PRD1. We show in this study that this unique vertex extends to the virus internal membrane via two integral membrane proteins, P20 and P22. These small membrane proteins are necessary for the binding of the putative packaging ATPase P9, via another capsid protein, P6, to the virus particle.