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Results for “Developmental stage”

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PubMed1990

Biotin as a probe of the surface of Ascaris suum developmental stages.

Sulfo-NHS-biotin (aqueous soluble) and NHS-biotin (organic soluble) labeled similar SDS-2ME (sodium dodecyl sulfate/beta-mercaptoethanol) soluble cuticular proteins of second stage larvae (L2) and third stage Ascaris suum larvae (L3). Comparable analysis of biotin-labeled fourth stage larvae (L4), young adults, and mature adult Ascaris suum revealed strong labeling of several SDS-2ME soluble cuticular proteins with NHS-biotin, while sulfo-NHS-biotin appeared to strongly label a single SDS-2ME soluble cuticular protein. Both biotin probes labeled only cuticular proteins, since no evidence of internal labeling was observed in any developmental stage examined by either electroblot analysis or by electron microscopy. Our data suggest a greater cuticular permeability to the organic soluble biotin reagent in the later developmental stages (greater than L3) of A. suum than to the aqueous soluble biotin reagent, and may indicate the presence of a hydrophobic barrier in the cuticle of the later stages of the parasite.

Animals↗

PubMed1998

Venous tone in the developmental stages of spontaneous hypertension.

The initial stages of hypertension in the spontaneously hypertensive rat (SHR) are characterized by an increase in cardiac output. Venous capacitance plays an important role in the control of cardiac output. This study tested the hypothesis that venous tone is elevated in the developmental stages of spontaneous hypertension. Male SHR or normotensive Wistar-Kyoto (WKY) rats were instrumented for the measurement of arterial pressure (FAP) and intrathoracic vena caval pressure (FVP). A latex-tipped catheter was advanced into the right atrium via the jugular vein. Mean circulatory filling pressure (MCFP), an index of integrated venomotor tone, was calculated as MCFP=FVP+(FAP-FVP)/VAR. FAP and FVP were recorded after 5 seconds of right atrial balloon inflation. The venous to arterial compliance ratio (VAR) was estimated as 76 for WKY and 106 for SHR. Mean arterial pressure (MAP), heart rate, and MCFP were recorded in conscious rats of 4 to 6 and 8 to 10 weeks of age. In 4- to 6-week-old rats, both MAP and MCFP were significantly elevated in the SHR (MAP, 129+/-6 mm Hg; MCFP, 6.6+/-0.4 mm Hg) compared with the age-matched WKY (MAP, 91+/-6 mm Hg; MCFP, 5.4+/-0.4 mm Hg), whereas heart rate was not significantly different. The elevations in MAP (SHR, 144+/-4 mm Hg; WKY, 102+/-3 mm Hg) and MCFP (SHR, 7.7+/-0.3 mm Hg; WKY, 6.0+/-0.2 mm Hg) in SHR were exaggerated at 8 to 10 weeks of age. After ganglionic blockade (chlorisondamine; 10 mg/kg), the differences in MCFP were no longer statistically significant between SHR and WKY at both 4 to 6 weeks of age (3.9+/-0.2 versus 4.0+/-0.3 mm Hg) and 8 to 10 weeks of age (5.0+/-0.3 versus 4.3+/-0.3 mm Hg, respectively). The differences in MAP at 4 to 6 weeks of age (79+/-7 versus 67+/-5 mm Hg, respectively) also were not statistically significant after ganglionic blockade. However, a significant difference in MAP between strains remained after ganglionic blockade in 8- to 10-week-old rats (90+/-5 versus 63+/-3 mm Hg, respectively). These findings indicate that venous tone is increased via autonomic effector systems during the developmental stages of spontaneous hypertension. These data also suggest that autonomic mechanisms predominate at very early stages, whereas nonautonomic mechanisms assume more importance in maintaining the elevated MAP as hypertension progresses.

Animals↗

PubMed1987

Levels and patterns of 125I-labeled transferrin binding in mouse embryonic teeth and kidneys at various developmental stages.

The iron-transporting serum glycoprotein, transferrin, is necessary for the cell proliferation, morphogenesis, and differentiation of mouse embryonic teeth and kidneys in organ culture. The stimulatory effect of transferrin is mediated by the binding of transferrin to its specific cell-surface receptor and by receptor-mediated endocytosis. Since, in both teeth and kidneys, the requirement for and responsiveness to transferrin depend on the developmental stage of the organ, we studied the binding of transferrin at various stages of tooth and kidney development by incubating tissues with 125I-labeled transferrin. The amount of bound transferrin was determined by measuring the tissue-incorporated radioactivity, and the binding sites were localized by autoradiography. During tooth development in vitro, the requirement for exogenous transferrin is lost as the teeth proceed from the early cap stage to the bell stage. The level of transferrin binding was found to decrease simultaneously, and in bell-stage teeth, the transferrin receptors were concentrated in the areas of most active cell proliferation. In kidneys, the number of transferrin receptors was highest at the stage during which the undifferentiated kidney mesenchyme becomes responsive to transferrin. These receptors were located in both the ureter epithelium and the metanephric mesenchyme, and they dramatically decreased in number with advancing kidney differentiation. The results of the present study indicate that, during the embryonic development of teeth and kidneys, the amount and localization of transferrin binding are correlated with cell proliferation. The number of transferrin receptors is highest during the developmental stages when cell proliferation is most active, and decreases with advancing differentiation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed2005

Effects of Developmental Stage and Sex on Lower Extremity Kinematics and Vertical Ground Reaction Forces During Landing.

Context: The presence or absence of biomechanical differences between the sexes before puberty may provide clues about the onset of adult landing pattern differences, which may help to explain the greater number of anterior cruciate ligament injuries in females than in males and provide the basis for interventions to reduce those injuries.Objective: To identify developmental sex-related and biomechanical differences during vertical jump landings.Design: A 2 x 2 developmental stage (prepubescent or postpubescent) x sex (male or female) between-subjects design.Setting: Controlled laboratory setting.Patients or Other Participants: Thirty prepubescent subjects (15 boys, age = 9.63 +/- 0.95 years; 15 girls, age = 9.19 +/- 1.00 years) and 28 postpubescent subjects (14 men, age = 23.57 +/- 3.23 years; 14 women, age = 24.22 +/- 2.27 years).Intervention: Subjects performed a vertical jump to a target set at 50% of their maximum vertical jump height ability.Main Outcome Measure(s): Hip and knee kinematics of the dominant lower extremity and vertical ground reaction forces during impact were analyzed.Results: We found significant main effects for developmental stage. Children demonstrated greater knee valgus and less hip flexion at initial contact and at maximum vertical force, less knee flexion at maximum vertical force, greater maximum vertical force and impulse, and a shorter time to maximum vertical force than the adults. No sex differences were found among the biomechanical variables measured.Conclusions: The presence of significant biomechanical differences between children and adults suggests that physical development influences landing patterns. Sex does not appear to influence landing patterns during a 50% maximum vertical jump landing. These findings add to the body of knowledge regarding developmental and sex comparisons in a functional landing task.

Journal Article↗

PubMed1993

Tumor induction by ras and myc oncogenes in fetal and neonatal brain: modulating effects of developmental stage and retroviral dose.

Introduction into fetal rat brain cells of a replication-defective retroviral vector harboring v-Ha-ras and v-gag-myc rapidly causes the induction of highly malignant undifferentiated neuroectodermal tumors following transplantation into the brains of syngeneic hosts [Wiestler, et al. (1992) Cancer Res. 52: 3760-3767]. In the present study, we have investigated the modulating effect of the developmental stage of neural target cells and of the dose of the retroviral vector used in the grafting experiments. Exposure of fetal cells from embryonic day (E)12 or E14 produced a 100% incidence of malignant neuroectodermal tumors which led to the death of recipient animals after a median latency period of 32 days. A 100-fold reduction of the virus dose from 2.062 x 10(6) to 2.062 x 10(4) focus-forming units/ml resulted in a lower tumor incidence of 25%. Of six neural grafts exposed to v-Ha-ras and v-myc at E16, only one showed evidence of tumorigenesis (low-grade astrocytoma and hemangioma). All other transplants were morphologically normal for observation periods of 26 weeks, indicating a marked loss of transforming activity of ras and myc in more advanced stages of brain development. In retrovirus-exposed donor cells which caused the development of neural tumors in recipient rats, malignant transformation was also evident during culture in vitro, usually after 9-12 days. Oncogene complementation was also studied in the newborn rat brain. After microinjection of the retroviral vector into the brain at postnatal day (P)0, P1 and P3, 5 out of 20 animals (25%) developed a total of seven brain tumors. Histopathologically, three of these neoplasms were malignant neuroectodermal tumors which, in contrast to those induced in fetal brain transplants showed evidence of focal glial and/or neuronal differentiation. In addition, we observed one oligodendroglioma, two hemangiomas and a malignant hemangioendothelioma. These data indicate that neural precursor cells and endothelia of the rat brain represent the major target cells for the complementary action of ras and myc and that the use of target cells from later developmental stages (E16 and postnatal) leads to the induction of both primitive and more differentiated neoplasms.

Animals↗

PubMed1987

High K+-mediated survival of spinal sensory neurons depends on developmental stage.

Elevated concentrations of K+ (35 mM) have previously been shown to support the survival of most embryonic chick sympathetic neurons in vitro (Wakade et al., Exp cell res 144 (1983) 377, [23]) and to be interchangeable with nerve growth factor (NGF) as a survival-promoting agent for these cells (Wakade & Thoenen, Neurosci lett 45 (1984) 71 [21]). In the present study, we show that dorsal root ganglion (DRG) neurons from embryonic day 6 do not survive in the presence of high K+, although both NGF and brain-derived neurotrophic factor (BDNF) each support the survival of more than 50% of the cells at this developmental stage. At E6, high K+ appears to have a cytotoxic effect on BDNF-dependent neurons, and there is also considerable inhibition of neurite outgrowth. At a later developmental stage (E12), high K+ supports the survival of about 40% of DRG cells. This subpopulation of neurons is distinct from that supported by NGF (as evidenced by the additivity of these two agents), but partially overlaps with that supported by BDNF (i.e., the two agents are less than additive). At E12, only approx. 20% of the cells can be supported by either NGF or BDNF, with the rest depending exclusively on one or the other of these factors. This is in contrast to the situation at E6, where there is considerable overlap between NGF- and BDNF-dependent populations.

Animals↗

PubMed2004

Molecular characterization of the glutamine synthetase gene in the Pacific oyster Crassostrea gigas: expression study in response to xenobiotic exposure and developmental stage.

In this study, we characterized the full-length cDNA and genomic sequence of the gene encoding cytosolic glutamine synthetase (CgGSII) in the Pacific oyster, Crassostrea gigas. A phylogenetic analysis of GS sequences showed that CgGS clustered with the invertebrate group as expected. We analyzed the expression of mRNA CgGSII using RT-PCR to follow the expression of this gene in gills and digestive gland of oysters exposed, under experimental conditions, to hypoxia and to several contaminants (hydrocarbons and two pesticide treatments, glyphosate and a mixture of atrazine, diuron and isoproturon). We also investigated the expression of CgGSII in different developmental stages of C. gigas. Our results show that CgGSII expression was highly regulated in xenobiotic-exposed oysters compared to the control for all the treatments. Likewise, CgGSII expression was highly regulated according to the developmental stage of C. gigas. Finally, use of CgGSII as a possible marker to monitor xenobiotic exposure in disturbed ecosystems is discussed.

Amino Acid Sequence↗

PubMed1989

Occupational allergy in an entomological research centre. II. Identification of IgE-binding proteins from developmental stages of the blowfly Lucilia cuprina and other species of adult flies.

Sera from 30 workers in an entomological research institute and from five Sydney asthmatics, all with serum IgE antibodies to sheep blowfly (Lucilia cuprina) antigens, were studied with the aim of identifying individual IgE-binding proteins in extracts of L. cuprina adult flies, larvae and eggs, and in extracts of related species of flies from the order Diptera. Using protein blotting, 21, 18 and nine different IgE-binding components were identified in extracts of L. cuprina adult flies, larvae and eggs, respectively. A component(s) of MW 67 kD found in all three developmental stages, showed the highest frequency of IgE-binding; reacting, for example, with 70% of the sera tested with the adult fly extract. Some components were detected in only one of the three developmental stages. Investigations of possible allergenic cross-reactivity between L. cuprina and extracts from six other related species of flies revealed IgE-binding bands in each of the extracts as well as in an extract of Agrotis infusa, a cutworm not belonging to the order Diptera. One strongly reacting component of MW 20 kD was detected in extracts of five different fly species as well as in the A. infusa extract. The results suggested that allergenic cross-reactivity between some fly species exists, and may extend to taxonomically unrelated insect species.

Animals↗

PubMed1996

Survival rates and sex ratio of bovine IVE embryos frozen at different developmental stages on day 7.

Two experiments were designed to determine the effects of stage of development on Day 7 of in vitro-produced bovine embryos on survival after deep freezing and on sex ratio. Bovine IVF embryos and bovine oviductal epithelial cells (BOEC) were co-cultured in TCM-199 and, on Day 7 after insemination (Day 0), were morphologically evaluated and divided into groups by developmental stage. In Experiment 1, embryos classified as early blastocysts, blastocysts and full-expanding blastocysts were randomly subdivided into 2 groups by replicate: 50% of the embryos were placed immediately in a new BOEC co-culture (fresh group), while the other 50% were frozen, thawed and placed in a new BOEC co-culture (frozen/thawed group). Embryos were frozen in 1.5 M glycerol using a standard slow cooling technique. Fresh and frozen/thawed embryos were compared for survival rate (embryos hatching/hatched) in BOEC co-culture over the following 3 d (i.e., Days 7 to 10). The overall survival of the 425 embryos (early to full-expanding blastocysts) was 33% and was not different between fresh (35%) and frozen/thawed (30%) embryos. Survival of embryos cultured fresh or after freezing/thawing was higher for full-expanding blastocysts than for early blastocysts or for blastocysts, both of which were not different. In Experiment 2, all frozen/thawed embryos used in Experiment 1 plus all morulae and hatched blastocysts collected and frozen on Day 7 without regard to survival were sexed utilizing the polymerase chain reaction (PCR) technique. Sex of the embryos, by stage of development on Day 7, was determined in order to compare the rate of development in BOEC co-culture with the sex ratio (percentage of males). A total of 235 embryos was sex-determined with an overall percentage of males of 51%, which was not different from the expected 1:1 sex ratio. Both full-expanding blastocysts and hatched blastocysts had a significantly higher (P < 0.05) proportion of males (68 and 100%, respectively), while morulae had a significantly lower proportion of males (24%). Early blastocysts and blastocysts did not differ from a 1:1 sex ratio. The results indicate that male embryos develop faster in vitro than female embryos. The higher survival rate of full-expanding blastocysts after freezing/thawing, and the production of a higher number of males than females among embryos of this developmental stage suggest that a greater number of male fetuses may result from the successful freezing and transfer of in vitro-produced bovine embryos.

Journal Article↗

PubMed2001

Identification of living oligodendrocyte developmental stages by fractal analysis of cell morphology.

The Mandelbrot's fractal dimension (D), a measure of shape complexity, has been used to quantify the complex morphology of living cells. Previous studies on glial cells have shown that as cells increase in morphological complexity, their "D" value increases, suggesting that "D" could be used to estimate their stage of differentiation. In the present study the box-counting method was used to calculate the "D" values of rat cerebellar oligodendrocytes during their differentiation in primary culture. These values were correlated with the immunoreactivity of cells to antigenic markers commonly used for assessing their stages of differentiation: A2B5, O4 and anti-galactocerebroside (Gal-C). Our results show that changes of the fractal dimension during differentiation follow the well known pattern of markers expression by these cells. These results demonstrate that A2B5-, O4-, and Gal-C-expressing oligodendrocytes can be confidently estimated from their respective fractal dimension values. Based on this immunocytochemical calibration, the calculation of "D" allows an easy and fast determination of the developmental stage of living (unstained) oligodendrocytes before the study of their physiological characteristics. Using this method we precisely identified living oligodendrocyte progenitors and early pro-oligodendrocytes expressing voltage-activated sodium currents that is a common characteristic of these two immature developmental stages (Sontheimer et al. [1989b] Neuron 2:1135-1145).

Aging↗

PubMed1988

Developmental-stage-dependent radiosensitivity of neural cells in the ventricular zone of telencephalon in mouse and rat fetuses.

Pregnant ICR mice were treated with single whole-body X-radiation at a dose of 0.24 Gy on day 10, 13, or 15 of gestation. Fetuses were obtained from mothers during 1 and 24 hours after irradiation. Pyknotic cells in the ventricular zone of telencephalon were counted in serial histological sections. Incidence of pyknotic cells peaked during 6 and 9 hours after irradiation in each gestation day group. Then, dose-response curves were obtained 6 hours after 0-0.48 Gy of irradiation. All three dose-response curves showed clear linearity in the dose range lower than 0.24 Gy. Ratios of radiosensitivity estimated from the slopes of dose-response curves in day 10, 13, and 15 groups were 1, 1.4, and 0.4, respectively. These demonstrated that ventricular cells in the day 13 fetal telencephalon were the most radiosensitive among the three different age groups. In order to confirm the presence of the highly radiosensitive stage common to mammalian cerebral cortical histogenesis, pregnant F344 rats were treated with single whole-body gamma-irradiation at a dose of 0.48 Gy on day 13, 14, 15, 17, or 19 of gestation. The incidence of pyknotic cells in the ventricular zone of telencephalon was examined microscopically during 1 and 24 hours after irradiation. The peak incidence was shown 6 hours after irradiation in all the treated groups, and the highest peak incidence was shown in day-15-treated group. The developmental stage of telencephalon of day 15 rat fetuses was comparable to that of day 13 mouse fetuses. Thus, the highest radiosensitivity in terms of acute cell death was shown in the same developmental stage of brain development, i.e., the beginning phase of cerebral cortical histogenesis, in both mice and rats.

Animals↗

PubMed1978

Developmental staging and thalidomide teratogenicity in the green monkey (Cercopithecus aethiops).

The developmental stages of 11 Cercopithecus aethiops embryos, 24 to 45 days post-mating, are described. The onset of organogenesis in this species is approximately five to seven days later than that reported for macaques and baboons. This temporal difference in the embryonic period is an important factor in the analysis of the subsequent teratogenic study. Oral administration of thalidomide in single or multiple (3-day) treatment periods to pregnant green monkeys between days 28 and 33 resulted in defects of the limbs which resemble those observed in macaques and baboons. However, the sensitive period occurs approximately four days later than that reported for another nonhuman primates. These results indicate that the sensitive period of the limbs to thalidomide coincides with their earliest development.

Abnormalities, Drug-Induced↗

PubMed2005

System, trends and perspectives of proteomics in dicot plants Part II: Proteomes of the complex developmental stages.

This review is devoted to the proteomes of the complex developmental stages of dicotyledoneous (dicot) plant materials. The two core technologies, two-dimensional gel electrophoresis (2-DGE) and mass spectrometry (MS), independently or in combination with each other, are propelling dicot plant proteomics to new discoveries and functions, with the establishment of tissue-specific and organelle proteomes, mostly in Arabidopsis thaliana and Medicago truncatula, revealing their complexity and specificity. These experimental proteomes have provided a good start towards the establishment of high-density 2-DGE reference maps and peptide mass fingerprint databases, for not only the model dicot plants, A. thaliana and M. truncatula, but also other important dicot plants, which will serve as a basis for proteomes of many other dicot plants and plant materials.

Arabidopsis↗

PubMed1984

Secretory IgA responses in rats to antigens of various developmental stages of Nippostrongylus brasiliensis.

Intestinal and faecal IgA responses against different developmental stages of Nippostrongylus brasiliensis were studied in Hooded Lister rats vaccinated subcutaneously with infective larvae or intragastrically with 4th-stage larvae, adult nematodes, adult worm somatic and metabolic antigens. It was found that intragastric administration of L4 or adult worms produced as high resistance to challenge as normal infection. Oral vaccination with adult nematode excretory-secretory (ES) and somatic antigens produced 69 and 17% protection respectively. All vaccination regimes induced a considerable IgA response against somatic and metabolic antigens of the intestinal stages. However, no correlation could be seen between the IgA levels and degree of protection. Analysis of intestinal mucosal, intestinal contents and faecal IgA specific to larval or adult antigens suggests that the IgA is probably induced by antigen(s) common to all intestinal stages. Also secretory IgA responses against surface antigens of intestinal L4 did not show stage specificity, although this was detected in sera.

Animals↗

PubMed1989

Molecular cloning and characterization of a Ca2+ + Mg2+-dependent adenosine triphosphatase from rat cardiac sarcoplasmic reticulum. Regulation of its expression by pressure overload and developmental stage.

To investigate the regulation of expression of cardiac Ca2+ + Mg2+-dependent ATPase (Ca2+-ATPase) in sarcoplasmic reticulum (SR), we isolated cDNA (pHA6) encoding a Ca2+-ATPase of rat cardiac SR. The clone consisted of 2,311 mRNA-derived nucleotides, which covered half the coding region and the entire 3'-untranslated regions. The nucleotides and deduced amino acid sequences of pHA6 showed striking homology, 89 and 98%, respectively, to those of rabbit Ca2+-ATPase of the slow-twitch form. Northern blot analyses revealed that the mRNA levels of Ca2+-ATPase were decreased by pressure overload and became 32% of sham in 1 mo. During the developmental stage the mRNA levels were very low in the fetal period and steeply increased around birth. These changes in mRNA levels were correlated with the corresponding protein levels. These results suggest that the expression of cardiac Ca2+-ATPase in SR is regulated by pressure overload and the developmental stage, at least in part, at the pretranslational level.

Aging↗

PubMed1997

Different pattern of interleukin-1 beta-(IL-1 beta), interleukin-1 receptor antagonist- (IL-1ra) and interleukin-1 receptor type I- (IL-1R tI) mRNA-expression in single preimplantation mouse embryos at various developmental stages.

The interleukin-1 (IL-1) system has been shown to play an important role in human and murine embryo implantation. Recent studies have documented immunohistochemical evidence of interleukin-1 beta (IL--1 beta), interleukin-1 receptor antagonist (IL-1ra) and interleukin-1 receptor type I (IL-1R tI) in human preimplantation embryos and protein levels of interleukin-1 alpha (IL-1 alpha), IL-1 beta and IL1ra in human preimplantation embryo culture fluid have been correlated with successful implantation and pregnancy. Our aim in this study was to detect IL-1 beta, Il-1ra and Il-1R tI mRNA in single preimplantation mouse embryos and to describe the frequency of positive mRNA-expression at different developmental stages. B6C3F1-mice, 12 weeks old were pregnant mare serum gonadotropin (PMSG)/human chorionic gonadotropin (hCG)-stimulated and mated. Animals were sacrificed at day 0.5, and zygotes were flushed from the tubes and cultured in HAMs-F10 medium. 2-cell- (2C-), 8-cell- (8C-), morula- (M-), early blastocyst- (EB-) and hatching blastocyst- (HB-) stage embryos were examined by one round of reverse transcriptase (RT) followed by two rounds of polymerase chain reaction (PCR) carried out on individual mouse embryos for beta-actin (internal standard), IL-1 beta, IL-1ra and IL-1R tI-mRNAs. The frequencies of positive mRNA-expressions were as follows (2C/8C/M/EB/HB); beta-actin: 91/96/100/100/98%; IL-1b: 0/0/2.5/6.25/19; IL-1ra; 0/5/30/41/74% and IL-1R tI: 0/0/10/20/25%. The incidence of IL-1ra mRNA expression increased with developmental stage. IL-1ra mRNA seems to be expressed in a very high percentage (74%) of embryos near the time of implantation, whereas the percentage of IL-1 beta-mRNA positive embryos is surprisingly low (19%).

Animals↗

PubMed2006

Developmental stages and germ cell lineage of the loach (Misgurnus anguillicaudatus).

The staging of embryonic and larval development, and the germ cell lineage of the loach, Misgurnus anguillicaudatus, are described. Fertilized eggs were obtained by artificial insemination. For the convenience of detailed observation and photography of the external appearance, we use dechorionated embryos. Through a series of operations, these embryos were cultured at 20 degrees C in an incubator. Embryonic and larval development of the loach was divided into five periods: cleavage, blastula, gastrula, segmentation, and hatching. Stages were assigned within each of these periods. Developmental stages were determined and named by morphological features and somite number. The staging series were photographed and tabulated. The germ cell lineage was then elucidated by whole mount in situ hybridization of mRNA expression of the germ-cell-specific marker vasa and histological analysis. Primordial germ cells (PGCs) of the loach derived from the cleavage furrows of 8-cell stage embryos began proliferation in the late blastula period and migrated to the gonadal anlagen through a migration pathway similar to that of the zebrafish. However, it is characteristic of the loach that PGCs migrate a long distance and stay in the posterior part of the yolk-extension region.

Animals↗

PubMed1999

Supplemental grant to continue cooperative agreements with 10 predictor variables grantees and their research coordinating center. CSAP, SAMHSA, HHS. Supplement to support an additional year of intervention follow-up and data collection among the Predictor Variables by Developmental Stage grantees and their Research Coordinating Center.

This notice is to inform the public that an estimated $1.65 million will be available to support up to 11 supplemental awards to 10 existing Predictor Variables Study Sites and one existing Research Coordinating Center in FY 1999. The purpose of the award is to support enhancement of current programs and allow the collection and analyses of additional follow-up data for children currently included in these studies. CSAP will make the awards based on the recommendations of the initial review group and the CSAP National Advisory Council. Supplemental awards will be made in Fiscal Year 1999, by September 30, 1999. The studies funded under this supplement are projected to end September 30, 2000. Eligibility is limited to existing SAMHSA/CSAP's Predictor Variables by Developmental Stage study sites, and their Research Coordinating Center. All currently active Predictor Variables grantees are eligible to apply for supplemental funds under this GFA. Given the short implementation time frame and limited funds available for this activity, as well as existing research protocols that limit the scope of new activities that could be introduced at this point in the study, the existing Predictor Variables projects are the only projects that can effectively implement the required booster sessions and follow-up data collection activities. These studies have already demonstrated that they can make a positive impact on children within selected developmental parameters. It is important to document that this impact on these same children can be maintained as they enter the next developmental stages. The Research Coordinating Center has put considerable effort into developing cross-site rapport and collecting process data from the individual sites; an effort that would be redundant and not cost-effective if attempted by another entity at this point in the project. Additionally, it is important to the continuity of the study that the Research Coordinating Center be able to continue its current analyses and be able to conduct secondary analyses based on the totality of the data submitted throughout the life of the study.

Child↗