bluuebunny/crossref_metadata_2025 huggingface dataset:
A Proposal for Case Definitions and Outcome Measures in Studies of Infantile Spasms and West Syndrome: Consensus Statement of the West Delphi Group
Authors: Lux Andrew L. , Osborne John P.
Published: October 2004

Summary: Purpose: To reach a broad consensus on case definitions, outcomes, and outcome measures that will ease future study design and facilitate comparison of data from different studies of infantile spasms and West syndrome.

Methods: Persons who had recently presented or published first‐author original research in this field were invited to participate in an e‐mail Delphi process and to invite other investigators or clinicians who they thought might participate.

Results: The process consisted of six rounds, anonymous except to the facilitator. In total, responses were received from 46 participants. The final statement was approved by 31 participants from 15 countries. It concluded that the primary clinical outcome,cessation of spasms, should denote absence of witnessed spasms from within 14 days of commencement of treatment, and for ≥28 consecutive days from the last witnessed spasm.Primary electroclinical outcomedenotes cessation of spasms with resolution of hypsarrhythmia.West syndromeshould be a defined subset of the syndrome of infantile spasms. Aninfantile spasms single‐spasm variantshould be recognized. Ways are suggested of handling subtle spasms in the context of clinical studies. It proposes a standard for reporting modifying and atypical features of hypsarrhythmia, a minimal set of baseline characteristics and outcomes that should be reported in trials of infantile spasms, and suggests a standard definition of relapse. Consensus was not reached on a definition of hypsarrhythmia.

Conclusions: We reached a clear consensus on many aspects of study design for the investigation of infantile spasms, although incomplete consensus was found on how to define EEG criteria.
Public Knowledge, Private Grief: A Study of Public Attitudes to Epilepsy in the United Kingdom and Implications for Stigma
Authors: Jacoby Ann , Gorry Joanne , Gamble Carrol , Baker Gus A.
Published: October 2004

Summary:  Purpose: For many people with epilepsy, the continuing social reality of their condition is as a stigma, thus representing a source of much private grief. To understand fully the nature of epilepsy stigma, it is important to examine attitudes and beliefs of not just the “targets” but also of the “perceivers” of stigma. Perceivers may hold erroneous beliefs and stereotypes that lead them to have negative expectations of people affected by epilepsy. This study examined levels of knowledge and attitudes among perceivers of epilepsy stigma in the U.K.

Methods: A random sample of >1,600 members of the general public was interviewed as part of the bimonthly UK Omnibus Survey. The response rate was 62%.

Results: One fourth of informants knew someone with epilepsy and one half had witnessed a seizure. One half agreed that people with epilepsy are treated differently by others, and themes of exclusion, restriction, and nonnormality were commonly cited as reasons for this. Epilepsy ranked second in a range of health conditions, in terms of which would cause them greatest concern if informants had to work with someone so affected. Responses to a series of attitude statements indicated that most held highly favorable attitudes; but one fifth agreed with the statement that people with epilepsy have more personality problems than do others. Responses were influenced by informants' sociodemographic characteristics.

Conclusions: The survey revealed attitudes and knowledge gaps that have the potential for discriminatory behavior. There are implications for how public education campaigns should be implemented, and who should be targeted.
Lipopolysaccharide‐induced Febrile Convulsions in the Rat: Short‐term Sequelae
Authors: Heida James G. , Boissé Lysa , Pittman Quentin J.
Published: October 2004

Summary:  Purpose: Febrile convulsions (FCs) occur in children as a result of fever. The mechanisms involved in the genesis of FCs and their long‐term consequences on brain development remain unclear. We have developed a model of FC, by using fever as a parameter, to test the hypothesis that fever can lower seizure threshold and to examine the neurologic sequelae of FCs.

Methods: Fourteen‐day‐old rat pups equipped with body‐temperature telemetry devices exhibited ∼1.5°C fevers after lipopolysaccharide (Escherichia coli, 200 μg/kg). During such fevers, concurrently administered doses of kainic acid that are normally subconvulsant were used to induce convulsions with fever. Animals were then killed at varying times for pathological and immunohistochemical studies.

Results: The pairing of lipopolysaccharide and subconvulsant kainic acid resulted in convulsions in ∼50% of febrile animals, with very low mortality. To study the neural correlates of these FCs, we used fos immunohistochemistry and found that animals with FCs had fos‐positive immunoreactivity in brain regions involved in seizures. After a period of 72 h, we also examined brains for pathologic changes and found no differences among our groups.

Conclusions: Our data indicate that a neuroimmune challenge and its accompanying fever reduce the seizure threshold. Furthermore, the FCs induced by fever in this model do not have short‐term adverse effects on the brain. In addition, this model, by incorporating physiologic fever, may be useful for examining the role of fever and its constituent mediators in the genesis of FCs.
Fracture Risk Associated with Use of Antiepileptic Drugs
Authors: Vestergaard Peter , Rejnmark Lars , Mosekilde Leif
Published: October 2004

Summary:  Purpose: To assess fracture risk associated with different antiepileptic drugs (AEDs). An increased fracture risk has been reported in patients with epilepsy. Classical AEDs have been associated with decreased bone mineral density. The effects of newer AEDs are unknown.

Methods: We undertook a population‐based pharmacoepidemiologic case–control study with any fracture as outcome and use of AEDs as exposure variables (124,655 fracture cases and 373,962 controls).

Results: All AEDs were associated with an increased fracture risk in an unadjusted analysis. After adjustment for prior fracture, use (ever) of corticosteroids, comorbidity, social variables, and diagnosis of epilepsy, carbamazepine [CBZ; odds ratio (OR), 1.18; 95% confidence interval (CI), 1.10–1.26], [and oxcarbazepine (OXC; 1.14, 1.03–1.26)], clonazepam (CZP; 1.27, 1.15–1.41), phenobarbital (PB; 1.79, 1.64–1.95), and valproate (VPA; 1.15, 1.05–1.26) were statistically significantly associated with risk of any fracture. Ethosuximide (0.75, 0.37–1.52), lamotrigine (1.04, 0.91–1.19), phenytoin (1.20, 1.00–1.43), primidone (1.18, 0.95–1.48), tiagabine (0.75, 0.40–1.41), topiramate (1.39, 0.99–1.96), and vigabatrin (0.93, 0.70–1.22) were not statistically significantly associated with fracture risk after adjustment for confounders. The relative increase was modest and in the same range for the significant and nonsignificant results. CBZ, PB, OXC, and VPA displayed a dose–response relation. Fracture risk was more increased by liver‐inducing AEDs (OR, 1.38; 95% CI, 1.31–1.45) than by noninducing AEDs (1.19; 95% CI, 1.11–1.27).

Conclusions: A very limited increased fracture risk is present in users of CBZ, CZP, OXC, PB, and VPA. A limited significant increase cannot be excluded for the other AEDs because of the statistical power.
Subjective Sleep Disturbance in Patients with Partial Epilepsy: A Questionnaire‐based Study on Prevalence and Impact on Quality of Life
Authors: De Weerd Al , De Haas Sanne , Otte Andreas , Trenité Dorothée Kasteleijn‐Nolst , Van Erp Gerard , Cohen Adam , De Kam Marieke , Van Gerven Joop
Published: October 2004

Summary:  Purpose: This study was designed to assess whether sleep disturbance is more frequent among patients with partial seizures and what impact on quality of life (QoL) sleep disturbance may have on patients with partial seizures.

Methods: Questionnaire booklets were mailed to 1,183 patients from four Dutch clinics. Each patient was asked to find two age‐ and gender‐matched controls to complete the same set of questionnaires [Sleep Diagnosis List (SDL), Medical Outcomes Study (MOS)‐Sleep Scale, Groningen Sleep Questionnaire, Epworth Sleepiness Scale, and the SF‐36 Health Survey]. The prevalence of sleep disturbance, based on the SDL, was compared between those with partial epilepsy and controls. Mean scores on sleep and the SF‐36 Physical (PCS) and Mental (MCS) Component Summary scales were compared.

Results: Responses from 486 patients and 492 controls were analyzed. Respondents with partial epilepsy had a highly significant, twofold higher prevalence of sleep disturbance compared with controls (38.6 vs. 18.0%; p < 0.0001). Most sleep‐disorder subscales showed significant abnormalities in respondents with epilepsy, compared with controls. Mean SF‐36 MCS and PCS scores were significantly lower in respondents with epilepsy compared with controls in both the strata with sleep disturbance and without (all p values <0.05). The presence of a sleep disturbance in respondents with epilepsy was associated with the greatest impairment in QoL.

Conclusions: Sleep disturbance is more than twice as prevalent in persons with partial epilepsy compared with controls, and most domains of sleep are significantly disturbed. Persons with partial epilepsy have significant QoL impairment, and sleep disturbance further compounds this.
Carbamazepine Enhances Discriminative Memory in a Rat Model of Epilepsy
Authors: Bernardi Rosane B. , Barros Helena M. T.
Published: October 2004

Summary:  Purpose: Seizures and antiepileptic drugs (AEDs) are the main causes for cognitive impairment in persons with epilepsy. It is still a matter of debate whether carbamazepine (CBZ) improves cognition because of its own psychotropic effects or because it is more effective to treat temporal epilepsy. Our objective was to analyze the performance of CBZ‐treated or nontreated pilocarpine epileptic rats in an object‐recognition test.

Methods: Twelve chronic pilocarpine‐induced epileptic rats were treated with CBZ, 40 mg/kg, or saline, t.i.d. for 8 days. Twenty‐one nonepileptic controls were treated with CBZ or saline. On day 8 of treatment, all rats were tested with an object‐recognition paradigm.

Results: No locomotor impairment was detected in chronic epilepsy or CBZ treatment, as exploration during training was not affected. Exploratory behaviors during the choice session were not decreased in rats treated with CBZ; therefore CBZ does not compromise procedural memory. Epileptic rats showed a nonsignificant change in the discrimination performance, and prolonged treatment with CBZ in epileptic rats induced a significant increase in object discrimination during the choice session.

Conclusions: Even though pilocarpine‐induced epileptic animals do not show compromised performance in the spontaneous object‐recognition test, prolonged CBZ treatment has a positive effect on a simple object‐discrimination task. These results may be associated with the psychotropic effects of CBZ.
The Nonmalformed Hemisphere Is Secondarily Impaired in Young Children with Hemimegalencephaly: A Pre‐ and Postsurgery Study with SPECT and EEG
Authors: Soufflet Christine , Bulteau Christine , Delalande Olivier , Pinton Florence , Jalin Claude , Plouin Perrine , Bahi‐Buisson Nadia , Dulac Olivier , Chiron Catherine
Published: October 2004

Summary:  Purpose: To study separately the functional value of each cerebral hemisphere in hemimegalencephaly (HME). HME is a unique model of unilateral hemispheric lesion, but one suspects that the non‐HME hemisphere also could be functionally impaired because the postsurgery outcome is less favorable than expected.

Methods: We performed simultaneous prolonged EEG and 133‐xenon SPECT (single‐photon emission computed tomography); we measured the absolute values of cerebral blood flow (CBF) in both hemispheres and compared them with the normal values previously acquired. Thirteen patients (aged 5–38 months) underwent 31 examinations, 20 before surgery (hemispherotomy) and 11 after.

Results: In the HME hemisphere, we confirmed the presurgical mixture of increased and decreased CBF due to intermittent ictal discharges. After surgery, CBF was decreased in most cases. In the non‐HME hemisphere, presurgery CBF was abnormal in 60% of the patients, increased and related mostly to diffuse interictal spikes on the same side, whereas normal CBF cases had focal spikes. After surgery, CBF was normal in 82% of cases, corresponding to an EEG without diffuse spikes. In the six patients longitudinally studied, CBF dramatically decreased after surgery in the HME hemisphere, whereas in the non‐HME hemisphere, CBF was mostly normal very early (three fourths before 2 months), increased as soon as 3 months, and normalized only after hemispherotomy, the more rapidly the child was operated on, the earlier it was.

Conclusions: This study shows that the function of the nonmalformed hemisphere is impaired as soon as the first months of the course of HME but can be restored after surgery. Our data support the recommendations to operate on the children as early as possible.
FLOW CYTOMETRY AND MICROSCOPY OF GAMETOGENESIS IN NITZSCHIA PUNGENS, A TOXIC, BLOOM‐FORMING, MARINE DIATOM1
Authors: Subba Rao Durvasula V. , Partensky Frédéric , Wohlgeschaffen Gary , Li William K. W.
Published: October 2004

The domoic acid‐producing diatom Nitzschia pungens Grunow f. multiseries Hasle, which is responsible for amnesic shellfish poisonings in Prince Edward Island, Canada, underwent gametogenesis when senescent cells (i.e. in stationary growth phase for more than 290 days) were subcultured into fresh FE medium and light intensity was increased from 33 to 530 μE · m−2· s−1. The number of gametes produced varied with the salinity of the medium, with a maximum at 23.5‰. Cells in the exponential growth phase (0.8 div · d−1) did not produce gametes, nor did senescent cells when transferred without change in light intensity. Anisogamous gametes, probably haploid, were isolated by combining conventional microscopy with flow cytometry. Zygotes resulting from syngamy yielded cigar‐shaped naviculoid cells, morphologically different from parent cells (heteromorphism). These cells, with a division rate of 1.9 div · d−1, could serve as a seed population and explain the origin and rapid progression of the toxic blooms of red‐water proportions that have been a public health problem in Eastern Canada. Production of domoic acid by postexponential and moribund cells but not by gametes, zygotes, or immediately resulting cells, provides an insight into the dependence of toxicity on the developmental history of this diatom.
DENSITY‐DEPENDENT GROWTH, ECOLOGICAL STRATEGIES, AND EFFECTS OF NUTRIENTS AND SHADING ON BENTHIC DIATOM SUCCESSION IN STREAMS1
Authors: Stevenson R. Jan , Peterson Christopher G. , Kirschtel David B. , King Christopher C. , Tuchman Nancy C.
Published: October 2004

The importance of immigration, growth, and competition for nutrients and light in benthic diatom succession was studied in experimental channels in a low‐nutrient stream. Diatom accumulation was greater in channels enriched with nitrate and phosphate (NP) than in control channels, reaching about 5 × 106 and 2 × 106 cells‐cm−2, respectively, after 30 d. Shading during late stages of community development reduced algal standing crop.

Synedra ulna (Nitz.) Ehr. and Achnanthes minutissima Kütz. were codominant during early stages of community development in both habitats, but succession to an A. minutissima‐dominated community was much faster in NP‐enriched than in control conditions. Species dominating early stages tended to immigrate quickly, whereas species that increased in relative abundance during community development had either fast growth rates or fast immigration and average growth rates.

Decreases in growth rates indicated resource supply became limiting during community development in control and enriched channels. Density‐dependent competition was indicated because nutrient concentrations in the water column and light did not decrease during the 30‐d study. Species autecologies were defined by effects of nutrient enrichment, shading, and community development on species growth rates. Differing autecologies of early and late succession species indicated that competition for nutrients was more important than competition for light.

Species autecologies also indicated ecological strategies. The species most stimulated by nutrient enrichment were least able to maintain growth rates as algal abundances on substrata increased. In addition, these species that best sustained their growth rates during succession tended to have the highest immigration rates, indicating that drift and immigration may have been an important mechanism of persistence for some populations when resources become limiting within thick benthic mats.
VARIATION IN NITROGEN PHYSIOLOGY AND GROWTH AMONG GEOGRAPHICALLY ISOLATED POPULATIONS OF THE GIANT KELP,MACROCYSTIS PYRIFERA(PHAEOPHYTA)1
Authors: Kopczak Charles D. , Zimmerman Richard C. , Kremer James N.
Published: October 2004

Three geographically isolated populations of the giant kelp,Macrocystis pyrifera(L.) C. Ag., were examined for responses to nitrate availability in batch culture experiments using juvenile sporophytes reared from spores in the laboratory. Although maximum rates of nitrate‐saturated growth were similar among groups, there were significant quantitative differences in the response to nitrate limitation that can be related to natural patterns of nutrient availability at these sites. Plants from Santa Catalina Island (most oligotrophic) achieved maximum growth rates at ambient nitrate concentrations that were lower than those for plants from Monterey Bay, California (most eutrophic), or Refugio State Beach (near Santa Barbara, California). Tissue nitrogen and amino acid concentrations were highest in plants cultured from Santa Catalina Island populations at all external nitrate concentrations, suggesting that differences in nitrate requirements for growth may reflect the efficiency of nitrate uptake and assimilation at subsaturating nitrate concentrations. Given the different physical environments from which these plants came, the data suggest that geographically isolated populations ofM. pyriferahave undergone genetic divergence that can be explained by ecotypic adaptation to unique habitat conditions at these sites.
NITROGENASE CONFINED TO RANDOMLY DISTRIBUTED TRICHOMES IN THE MARINE CYANOBACTERIUM TRICHODESMIUM THIEBAUTII1
Authors: Bergman Birgitta , Carpenter Edward J.
Published: October 2004

Nitrogenase reductase (Fe‐protein) was detected in the marine planktonic cyanobacterium Trichodesmium. The molecular weight was about 38 kD, as shown by western blotting using anti ‐Rhodospirillum rubrum nitrogenase reductase antiserum. The enzyme was confined to a limited number (ca. 10–40%) of randomly distributed trichomes in the Trichodesmium colonies, as shown by immunogold localization and transmission electron microscopy. Associated microorganisms had little or no nitrogenase. Nitrogenase showed a diel cycle in localization: present throughout the cytoplasm of cells in N2‐fixing (daytime) colonies but at the periphery of non‐N2‐fixing (nighttime) colonies. This structural arrangement of N2‐fixing trichomes and nitrogenase is novel and different from the previously held paradigm for this and other diazotrophic cyanobacteria.
RESPONSE OF MARINE SYNECHOCOCCUS (CYANOPHYCEAE) CULTURES TO IRON NUTRITION1
Authors: Rueter John G. , Unsworth Nancy L.
Published: October 2004

Three clones of marine Synechococcus (WH6501, WH7803, and WH8018) were grown through at least three transfers, at 6‐day intervals, in synthetic medium with total iron concentrations from 10−9 to 10−6 M. After 6 days of exponential growth, these cultures were harvested, and the cell density and protein and pigment concentrations were measured. Aliquots of the culture were assayed for their carbon fixation rates at two light intensities. Cell density and protein concentration increased by up to 7.8 times over a range of iron from the lowest (10−9 M) to the highest concentrations (10−6 M). The concentration of chlorophyll‐a and phycobiliproteins showed a wider range of response, increasing by up to 48 times. The carbon fixation rate (per mL of culture) also increased approximately 40 times over the total range of iron concentration. The ranges of these biochemical and physiological responses were much lower than the range of total available iron, which was 1000‐fold, and the range of total cellular iron, which was estimated to be about 160‐fold. This “less‐than‐linear” relationship indicates that the cells are adapting to make more efficient use of iron under limiting conditions. Our results demonstrate characteristics of iron‐limited Synechococcus that may be important in understanding the relationships between primary productivity and iron availability in the oceans.
IMMUNOFLUORESCENCE MICROSCOPY OF FERTILIZATION AND PARTHENOGENESIS IN LAMINARIA ANGUSTATA (PHAEOPHYTA)1
Authors: Motomura Taizo
Published: October 2004

Normal fertilization and parthenogenesis of unfertilized eggs were observed in Laminaria angustata Kjellman by indirect immunofluorescence microscopy using a tubulin antibody. Sperm aster formation did not occur at plasmogamy. The centrosome of the egg gradually disappeared. Shortly after karyogamy, one centrosome reappeared near the zygote nucleus. During mitosis, the centrosome replicated and the daughter centrosomes migrated to opposite poles. The mitotic spindle was formed by microtubules that elongated from both poles. After the first cell division, each of the daughter cells received one centrosome that persisted throughout the development of the sporophyte. During parthenogenetic development, abnormal mono‐, tri‐, and multi‐polar spindles were formed. These abnormal spindles caused abnormal nuclear and cytoplasmic division. Thus, cells were produced with 1) no nuclei, 2) multiple nuclei, 3) irregular numbers of chromosomes, and/or 4) no centrosomes. This is one of the reasons for the abortion and abnormal morphogenesis during parthenogenesis. Ultrastructural observations showed that, although cells of some parthogenetic sporophytes have centrioles, cells of almost all abnormally shaped parthenogenetic sporophytes lack centrioles. These results suggest that centrioles are required for normal centrosomal functions in Laminaria. Although centrioles are inherited paternally, some centrosomal material appears to be present or produced de novo in unfertilized eggs.
KARYOLOGY OF BANGIA ATROPURPUREA (RHODOPHYTA, BANGIALES) FROM MEDITERRANEAN AND NORTHEASTERN ATLANTIC POPULATIONS1
Authors: Gargiulo Gaetano M. , De Masi Flora , Tripodi Giacomo
Published: October 2004

We studied the karyology of Bangia atropurpurea (Roth) C. Ag. collected from marine and freshwater populations from the Mediterranean region and some northeastern Atlantic localities. Gametophytic thalli had two haploid karyotypes, n = 3 and n = 4. The n = 4 karyotype was only occasionally present in the Mediterranean and was also found in one Atlantic population, confirming a previous report. We propose that the four‐chromosome karyotype is an aneuploid form, n + 1. Chromosomes were frequently observed either in a parallel arrangement or in a circular configuration.
REGENERATION AND SEXUAL DIFFERENTIATION OF GRIFFITHSIA JAPONICA (CERAMIACEAE, RHODOPHYTA) THROUGH SOMATIC CELL FUSION1
Authors: Hwang Mi Sook , Kim Hyung‐Seop , Lee In Kyu
Published: October 2004

Hybrid cells were obtained from somatic cell fusion among male, female, and tetrasporangial plants in Griffithsia japonica Okamura by a wound‐healing process. Isolated fusion cells regenerated new mature plants with mixed reproductive structures. The plants regenerated from hybrid cells between male and female plants developed into 1) spermatangiate, 2) carpogonial, 3) bisexual with spermatangia and carpogonial branches, 4) mixed‐phase with spermatangia and tetrasporangia, or 5) bisexual/mixed‐phase plants with spermatangia, carpogonial branches, and tetrasporangia. About 70% of the plants regenerated from hybrid cells between male and female plants produced tetrasporangia that were always formed with spermatangia on a single cell. Some of those tetrasporangia released tetraspores, six of which gave rise to mature plants. The plants regenerated from hybrid cells between male and tetrasporangial plants developed into spermatangiate, tetrasporangiate, or mixed‐phase plants with spermatangia and tetrasporangia. The plants regenerated from hybrid cells between female and tetrasporangial plants developed into carpogonial, tetrasporangiate, or mixed‐phase plants with carpogonial branches and tetrasporangia. All types of reproductive structures we re functional.
INFLUENCE OF CELL VOLUME ON THE GROWTH AND SIZE REDUCTION OF MARINE AND ESTUARINE DIATOMS1
Authors: Mizuno Makoto
Published: October 2004

The maximal growth rate (μmax) of 19 marine and estuarine diatoms decreased with increasing cell volume (V). The relationship between log μmax(Y) and log V (X) was calculated. Statistical analyses showed that the slope of the equation was not significantly different from those obtained by other researchers and that the 95% confidence intervals of mean μmaxat cell volumes of 103–105μm3were not significantly different from those cited in most studies. A new regression line for diatoms was calculated as follows: log μmax= 0.47–0.14 log V; r =–0.69.

The rate of size reduction per generation of the 19 diatom species ranged from 0.03 to 0.87 μm per generation. The rate increased with increasing cell length and cell volume and with decreasing maximum division rate. Statistical analyses showed that the rate was closely related to the cell volume and to the reciprocal of the growth rate. The relationships between maximal growth rate and cell volume and between rate of size reduction and cell volume showed that a diatom with a large volume had a smaller maximal growth rate and a larger rate of size reduction than a diatom with a small volume. The estimates using the equation for the regression line between the rate of size reduction and the reciprocal of maximum division rate indicated that a diatom with a high maximum division rate would need more generation equivalents for a certain size reduction than a diatom with a low maximum division rate, but the periods required for reduction would be approximately equal irrespective of maximum division rate.
ANTINEOPLASTIC ACTIVITY OF CULTURED BLUE‐GREEN ALGAE (CYANOPHYTA)1
Authors: Patterson Gregory M. L. , Baldwin Cynthia L. , Bolis Christine M. , Caplan Faith R. , Karuso Helen , Larsen Linda K. , Levine Ira A. , Moore Richard E. , Nelson Carrie S. , Tschappat Kathryn D. , Tuang Grace D. , Furusawa Eiichi , Furusawa Shinobu , Norton Ted R. , Raybourne Richard B.
Published: October 2004

A large‐scale screening program was initiated to evaluate laboratory‐cultured blue‐green algae (cyanobacteria) as a source of novel antineoplastic agents. Approximately 1000 cyanophyte strains from diverse habitats were cultured to provide extracts for testing. The screening program identified the families Scytonemataceae and Stigonemataceae as prolific producers of novel cytotoxic compounds. Rates of rediscovery of known compounds were relatively low.
cAMP AND cAMP‐STIMULATED PROTEIN PHOSPHORYLATION IN ZOOSPORES OF BROWN ALGAE1
Authors: Ohmori Masayuki , Muto Shoshi , Miyachi Shigetoh
Published: October 2004

Adenosine 3′,5′‐cyclic monophosphate (cAMP) and guanosine 3′,5′‐cyclic monophosphate (cGMP) were detected at concentrations of 8–11 and 10–20 pmol · mg−1 protein, respectively, in zoospores of a brown alga, Undaria pinnatifida (Harvey) Suringer. Cellular levels of these cyclic nucleotides did not substantially change during dark to light transition. cAMP‐stimulated protein phosphorylation was found in soluble cell‐free extracts of zoospores of Undaria pinnatifida and Laminaria angustata Kjellman.
STEROLS OF CEPHALEUROS (TRENTEPOHLIACEAE), A PARASITIC GREEN ALGA1
Authors: Patterson Glenn W. , VanValkenburg Shirley D.
Published: October 2004

24‐Ethylcholesterol, 24‐ethylcholesta‐5,7,22‐trienol, dihydrolanosterol, 24‐ethylcholesta‐7,22‐dienol, and 4‐methyl‐24‐ethylcholesta‐7,22‐dienol were identified in cultured Cephaleuros in small quantities. Cholesterol made up 19% of the total sterol. The principal sterol, making up 65% of the total sterol, was 4,24‐dimethylcholest‐7‐enol, a new algal sterol. This is the first report of this sterol as the principal sterol of any living organism.
A NOVEL TECHNIQUE FOR PREPARATION OF AXENIC CULTURES OF SYMBIODINIUM (PYRROPHYTA) THROUGH SELECTIVE DIGESTION BY AMOEBAE1
Authors: Polne‐Fuller Miriam
Published: October 2004

The marine amoeba Trichosphaerium Am‐I‐7 was used as a tool for preparing unialgal axenic cultures of nondigestible Symbiodinium and Porphyridium species. The resistance of these unicellular algae to the amoebal digestive enzymes, and the differential digestion of bacteria, protozoans, and other algae, resulted in cleansed cells of Symbiodinium and Porphyridium that remained in the amoebal food vacuoles. During multiple fission, the amoeba evacuated its food vacuoles and released the trapped and intact algae, which were then successfully cultured. This method of cleaning was especially useful with algal species that were sensitive to antibiotics or other germicidal agents.
PHAGOTROPHY IN THE FRESHWATER, PHOTOSYNTHETIC DINOFLAGELLATE AMPHIDINIUM CRYOPHILUM1
Authors: Wilcox Lee W. , Wedemayer Gary J.
Published: October 2004

The cold‐water, photosynthetic dinoflagellate Amphidinium cryophilum Wedemayer, Wilcox & Graham feeds phagotrophically on other dinoflagellate species. Food is ingested through a feeding tube, termed here the “phagopod,” which extends from the antapex. The peduncle of this organism plays no observable role in the feeding process. The phagopod is essentially a hollow cylinder composed electron‐opaque material that is possibly deposited on a membrane. No Amphidinium cytoplasmic components, including microtubules or other cytoskeletal elements, were observed in the phagopod. Prefeeding cells aggregate, in small clumps near prey organisms with their phagopods extended. Eventually some cells commence feeding, first inserting the phagopod through the prey cell‐covering and then slowly, over a period of 10 min or more, drawing cytoplasm through the phagopod and into a nascent food vacuole. Both light and electron microscopy suggest that one or more prey cell amphiesmal membranes remain intact during the feeding process. Upon completion of feeding, the Amphidinium cell swims off with a prominent food vacuole in the hypocone, leaving at least part of the phagopod attached to the prey cell. Phagotrophy in A. cryophilum seems to vary with light intensity. At low light intensities, cells feed phagotrophically and are nearly colorless, whereas at high light levels they feed much less frequently, if at all, and are brightly pigmented.
INTRON REVEALED BY NUCLEOTIDE SEQUENCE OF LARGE SUBUNIT OF RIBULOSE‐1,5‐BISPHOSPHATE CARBOXYLASE/OXYGENASE FROM CODIUM FRAGILE (CHLOROPHYTA): PHYLOGENETIC ANALYSIS1
Authors: Manhart James R. , VonderHaar Raymond A.
Published: October 2004

The coding sequence for the large subunit of ribulose‐1,5‐bisphosphate carboxylase/oxygenase (rbcL) from Codium fragile (Suringar) Hariot chloroplast DNA is 1428 bp in length and contains a 1813‐bp group II intron. The only other organisms in which introns have been found in the rbcL gene are Euglena and Astasia. The Codium intron likely had a separate origin from the Euglena and Astasia introns, based on comparisons of intron sizes and sequences. Phylogenetic analyses of rbcL nucleotide and amino acid sequences place Codium between Chlorella and two Chlamydomonas spp., indicating that the Chlorophyceae may be polyphyletic.
EVOLUTIONARY RELATIONSHIPS BETWEEN FOUR SPECIES OF CLADOPHORA (CLADOPHORALES, CHLOROPHYTA) BASED ON DNA–DNA HYBRIDIZATION1
Authors: Bot Peter V. M. , Brussaard Corina P. D. , Stam Wytze T. , van den Hoek Chris
Published: October 2004

Analysis of the reassociation kinetics of the DNA from Cladophora pellucida (Huds.) Kütz. indicates that the genome of this benthic alga is comprised of approximately 75% repetitive sequences. Single‐copy sequences reassociated with a rate constant of 1.8 × 10−3 M−1· s−1, which corresponds to a haploid genome size of 4.7 × 108 bp. Genotypic relationships between members of the form section Longiarticulatae were determined by the method of DNA–DNA hybridization. No significant divergence was observed between the single‐copy sequences of C. pellucida isolates from the East Atlantic coast and Mediterranean Sea. Cladophora feredayi Harv. and C. att. ad pellucida from Australia and C. pellucidoidea van den Hoek from the West Atlantic coast were highly and about equally divergent from C. pellucida. The data support the hypothesis that the West Atlantic–West Pacific divergence reflects the middle Miocene closure of the Mediterranean–Indo‐Pacific seaways, and the hypothesis that the Northwest Atlantic–Northeast Atlantic divergence reflects the middle Miocene thermal separation of these coasts.
RESTRICTION ENDONUCLEASE ANALYSIS OF RIBOSOMAL DNA SEQUENCE VARIATION IN LAMINARIA (PHAEOPHYTA)1
Authors: Bhattacharya Debashish , Mayes Charlene , Druehl Louis D.
Published: October 2004

The taxonomy and evolutionary relationships of species in the genus Laminaria are poorly understood. Previous studies have demonstrated significant plasticity of morphological characters used to describe taxa, and interfertility has been reported among putative species. We analyzed nuclear ribosomal DNA (rDNA) sequence variation in eight species of Laminaria (L. agardhii Kjell., L. digitata (Huds.) Lamour., L. groenlandica Rosenv. [sensu Druehl 1968], L. longicruris De la Pyl., L. longipes Bory, L. saccharina (L.) Lamour., L. setchellii Silva, and L. yezoensis Miyabe) to elucidate evolutionary relationships in this genus. Restriction maps were constructed using a small subunit rDNA probe from Costaria costata (Turn.) Saunders, an rDNA repeat from the nematode Caenorhabditis elegans, and 11 hexameric restriction endonucleases in an annealing analysis of genomic DNA. Laminaria rDNA restriction maps were compared to each other and to that of the outgroup taxon, C. costata. rDNA restriction maps of Laminaria species and C. costata were similar. Restriction fragment length polymorphisms mapped to both the coding regions and the nontranscribed spacer of rDNA. Laminaria species were distinguished with this method. The restriction maps of L. agardhii, L. saccharina, and L. longicruris were identical, supporting a previous hypothesis that these species are conspecific. Comparison of restriction maps of Laminaria species suggested that the generic subdivision of Sections Simplices and Digitatae may be invalid.
INORGANIC CARBON LIMITATION OF PHOTOSYNTHESIS IN ULVA ROTUNDATA (CHLOROPHYTA)1
Authors: Levavassur Guy , Edwards Gerald E. , Osmond C. Barry , Ramus Joseph
Published: October 2004

A computerized oxygen electrode Astern was used to make rapid and accurate measurements of photosynthetic light and dissolved inorganic carbon (DIC) response cures with a macroalga. Ulva rotundata Blid. was grown in an outdoor, continuous flow system in seawater under sunlight or 9% of sunlight at Beaufort, North Carolina. The light compensation points in the shade‐ and sun‐grown plants, measured in seawater, were at photon flux densities (PFDs) of 16 and 27 μmol. Photons·m−2·s−1, respectively but the quantum yield of O2 evolution was not significantly different. Rates of photosynthesis in seawater per unit area of thallus under saturating light and rates of dark respiration were about 1.5‐fold higher in sun‐ than in shade‐grown plants. The concentration of DIC in seawater (approximately 2 mM) limited photosynthesis at absorbed PFDs above 60–70 μmol photons·m−2·s−1 Addition of 20 mM inorganic carbon had no effect on quantum yield but caused about a 1.5‐fold increase in the light‐saturated photosynthetic rate in both shade‐ and sun‐grown Ulva. The effect of DIC supplementation was greatest in plants grown in October and least in plants grown in June. The light‐ and DIC‐saturated rate of photosynthesis in seawater was similar to the maximum rate obtained by exposing Ulva to 10% CO2, in the gas phase. The carbon isotope values (δ13C, reflecting the 13C/12C ratio compared to a standard) of Ulva grown in the same seawater supply were dependent on light and agitation. Samples from Beaufort Inlet were more negative (δ13C value, −20.03‰) than those grown in bright light with agitation (δ13C value, −17.78‰ outdoors; −17.23‰ indoors), which may indicate DIC supply limited carbon uptake in seawater.
NITROGENASE ACTIVITY, PHOTOSYNTHESIS, AND THE DEGREE OF HETEROCYST AGGREGATION IN THE CYANOBACTERIUM ANABAENA FLOS‐AQUAE1
Authors: Kangatharalingam N. , Dodds Walter K. , Priscu John C. , Paerl Hans W.
Published: October 2004

The nitrogen‐fixing cyanobacterium Anabaena flosaquae Lyngb.) De Breb. exhibited aggregation of heterocysts from different filaments in a eutrophic lake and when grown in unialgal culture. The resulting aggregated filaments formed unialgal flocculent masses having a thickness of several centimeters that apparently resulted from cohesive mucilage surrounding heterocysts. We tested the effects of heterocyst aggregation on nitrogenase activity (NA) and photosynthesis in relation to microscale environmental O2 gradients. The redox indicator 2,3,5‐triphenyl tetrazolium chloride showed that aggregated heterocysts had lower intracellular redox potential than those that were dispersed. Microelectrode measurements showed that heterocyst aggregates in actively photosynthesizing flocculent masses were surrounded by a microzone of O2 30% higher than in the surrounding water: dispersed cells exhibited no such elevated O2 microzone. Despite high levels of O2, NA was greater in aggregated than dispersed samples, Microscale irradiance measurements made with a fiber optic light probe showed that 40% of the incident light was absorbed within the first 3 mm of a 1‐cm‐thick flocculent mass. The microscale irradiance data, together with nitrogenase and photosynthesis versus irradiance data, imply that the ratio of N:C fixation is lowest in filaments on the outside of 1.5–2.0‐cm masses and increases toward the center.
FEEDING APPARATUS OF THE COLORLESS FLAGELLATE KATABLEPHARIS (CRYPTOPHYCEAE)1
Authors: Lee Robert Edward , Kugrens Paul , Mylnikov Alexander Petrovich
Published: October 2004

The feeding apparatus of Kalablepharis ovalis (isolated from a freshwater impoundment in Colorado) and Katablepharis clone G‐2 (isolated from the littoral of the Black Sea near Yalta in the Crimea) consists of inner and outer oval‐shaped arrays of microtubules that begin at the anterior end of the cell and pass into the posterior of the cell. Each array of microtubules contains groups of microtubules with two to eight microtubules per group depending on the position of the array in the cell. A specialized area of the plasma membrane, the mouth, occurs at the anterior end of the cell. The mouth is oval with the long axis oriented dorsoventrally and consists of a raised ridge surrounding a central depression. The anterior end of the microtubules of the inner and outer arrays supports the raised ridge of the mouth. In freeze‐fracture replicas, the protoplasmic face of the plasma membrane contains intramembrane particles on the raised ridge of the mouth. Three small membrane‐cisternae occur on the protoplasmic side of the plasma membrane in the area of the mouth. Katablepharis clone G‐2 also has five or six additional large membrane‐cisternae associated with the inner microtubular array in the anterior portion of the cell. These larger membrane‐cisternae do not occur in K. ovalis. Vesicles with electron‐dense contents occur in association with the microtubular arrays. Katablepharis ovalis has a second type of vesicle containing a single‐membrane profile associated with the microtubule arrays. The structure of the microtubular arrays in Katablepharis is compared with similar structures in suctorian ciliates and dinoflagellates.
NUCLEOTIDE SEQUENCE OF THE GENES FOR RIBULOSE‐1,5‐BISPHOSPHATE CARBOXYLASE/OXYGENASE LARGE SUBUNIT AND RIBOSOMAL PROTEIN S14 FROM A CHLORELLA‐LIKE GREEN ALGA1
Authors: Amberg Sean M. , Meints Russel H.
Published: October 2004

Two chloroplast genes were sequenced from an exsymbiotic strain of a eukaryotic, Chlorella‐like green alga. The genes for the large subunit of ribulose‐1,5‐bisphosphate carboxylase/oxygenase (rbcL) and the ribosomal protein S14 (rps14) were oriented in the same direction and were separated by 402 bp. The rbcLs of the exsymbiont and a free living Chlorella ellipsoidea were compared with other reported rbcL sequences. The rbcL gene of the exsymbiont is closely related to that of free‐living Chlorella ellipsoidea. This is the first published report of an rps 14 gene sequence from an alga.
NEW ENDOLITHIC CYANOBACTERIA FROM THE ARABIAN GULF. I.HYELLA IMMANISSP. NOV.1
Authors: Al‐Thukair Assad A. , Golubic Stjepko
Published: October 2004

Hyella immanis, a new species of endolithic cyanobacterium that penetrates carbonate ooid sand grains in the Arabian/Persian Gulf, is formally described. Natural populations of the new taxon were sampled in moving ooid shoals at four locations along the east coast of Saudi Arabia, together with seven other endolithic taxa. The new species was isolated, and its properties were studied under experimental conditions. Small reproductive cells (baeocytes) exhibited positively phototactic gliding motility following release. In culture they grew into colonies forming isodiametric packages (prevalent on agar) and distinct pseudofilaments (prevalent in liquid culture). Carbonate penetration of the cultured strain in liquid culture proceeded at a rate of up to 10 μ· d−1. Agitation of cultures with magnetic stirrers enhanced the frequency of borings and the initial boring rates, but it had no effect on the continuing boring activity. A fossil counterpart of the new species was identified in Upper Proterozoic (700–800 million years old) silicified oolitic and pisolitic rocks of East Greenland.
SEXUAL REPRODUCTION IN STEPHANODISCUS NIAGARAE (BACILLARIOPHYTA)1
Authors: Edlund Mark B. , Stoermer Eugene F.
Published: October 2004

We observed sexual reproduction in a clonal culture of Stephanodiscus niagarae Ehrenb. and used light and scanning electron microscopy to absent flagellated male cells, auxospore growth, initial valve structure and production, and subsequent daughter cell division. Free auxospores were spherical and nonsiliceous throughout growth, producing hemispherical initial valves devoid of spines and with nonfasciculate striae. Pregametangial cells averaged 43% of the diameter of the daughter cell population and were 1/9 the biovolume of initial, cells. This paper is the first confirmed report of sexual reproduction in S. niagarae, although it appears that specimens of Actinocyclus niagarae H. L. Smith, described from Lake Erie in 1878, are actually initial valves of S. niagarae.
Evaluation of an injection depot formulation of buprenorphine: placebo comparison
Authors: Sigmon Stacey C. , Wong Conrad J. , Chausmer Allison L. , Liebson Ira A. , Bigelow George E.
Published: October 2004

Aims  Buprenorphine is a mu‐opioid partial agonist that is marketed in a sublingual formulation as a treatment for opioid dependence. A microcapsule depot sustained‐release formulation has been developed which may offer effective treatment  of  opioid  dependence  while  also  minimizing  risks  of  illicit  diversion or patient non‐compliance. The present study examined the efficacy of depot buprenorphine in suppressing the opioid withdrawal syndrome and in attenuating the effects of exogenous opioid challenge.

Design  A placebo‐controlled, double‐blind, randomized trial.

Setting  A closed residential research facility.

Participants  A total of 15 opioid‐dependent participants were enrolled into the 6‐week study.

Intervention  Fifteen participants were randomized to receive a single subcutaneous depot injection containing buprenorphine (58 mg) or placebo. Two participants, both of whom received placebo, terminated participation after depot administration. Thirteen participants (six buprenorphine, seven placebo) completed the 6‐week study and were assessed throughout the study for signs and symptoms of opioid withdrawal and for response to weekly subcutaneous challenges with 3 mg hydromorphone.

Measurement  Subjective, physiological and observer‐rated indices of opioid withdrawal and opioid agonist effects.

Findings  Depot buprenorphine provided more effective relief from opioid withdrawal than placebo, as evidenced by significantly fewer buprenorphine participants requiring supplemental medications for withdrawal suppression after depot administration compared to participants receiving placebo. In the weekly hydromorphone challenge sessions, depot buprenorphine significantly reduced opioid response on measures of subjective effects and pupillary diameter.

Conclusions  Results from this double‐blind, placebo‐controlled study indicate that depot buprenorphine is effective in providing both withdrawal suppression and opioid blockade. Future studies examining additional doses and repeated dosing regimens with depot buprenorphine are warranted.
Point mutations of 3BP2 identified in human‐inherited disease cherubism result in the loss of function
Authors: Miah S. M. Shahjahan , Hatani Tomoko , Qu Xiujuan , Yamamura Hirohei , Sada Kiyonao
Published: October 2004

Adaptor protein 3BP2 positively regulates the high affinity IgE receptor (FcɛRI)‐mediated activation of degranulation in mast cells. Genetic study identified the point mutations of 3BP2 gene in human‐inherited disease cherubism. The multiple cysts in cherubism lesion of jaw bones are filled with the activated osteoclasts and stromal cells, including mast cells. By over‐expression study using rat basophilic leukaemia RBL‐2H3 mast cells, we have analysed the effect of the point mutations on the function of 3BP2 protein, which plays a positive regulatory role on FcɛRI‐mediated mast cell activation. Over‐expression of 3BP2 mutants suppressed the antigen‐induced degranulation and cytokine gene transcription. Antigen‐induced phosphorylation of Vav1, activation of Rac1, extracellular signal regulated kinase (ERK), c‐Jun N‐terminal kinase (JNK), p38 mitogen activated protein kinase (MAPK), inhibitor of nuclear factor κB kinase (IKK) and nuclear factor of activated T cells (NFAT) were all impaired in the cells over‐expressing the cherubism mutants of 3BP2. Furthermore, cherubism mutations of 3BP2 may abrogate the binding ability to interact with chaperone protein 14‐3‐3. These results demonstrate that over‐expression of the mutant form of 3BP2 inhibits the antigen‐induced mast cell activation. It suggests that point mutations of 3BP2 gene cause the dysfunction of 3BP2 in vivo.