bluuebunny/crossref_metadata_2025_split huggingface dataset:
Genetic Manipulation of Mammalian Cells by Microinjection: Figure 1.
Authors: Rose David W. 
Published: May 2007

INTRODUCTIONMicroinjection allows the introduction of molecules into a defined population of cells at a defined concentration, and the timing of the experiment can be controlled stringently, minimizing problems associated with overexpression. Delivery by microinjection can be used for any type of cell that is adherent in culture, including primary cells. Perhaps its most powerful aspect is the ability to introduce several types of reagents into cells simultaneously, including DNA constructs, a labeled dextran to mark injected cells, antibodies, short interfering RNAs (siRNAs), and peptides. Because siRNA can be generated easily and rapidly for any target gene, it is relatively simple to assess many effects of the knockout of any gene in any type of adherent cell in a matter of days. However, microinjection is not trivial to learn, and requires attention to detail. Also, the limited number of cells involved often does not permit the subsequent analysis of effects on some biochemical parameters. While the major limitation of the approach is the small amount of material obtained, as the sensitivity of analysis techniques increases, so does the usefulness of this method. We emphasize here recent advances in the use of microinjection in combination with siRNA technology.
Embryo Dissection and Micromanipulation Tools: Figure 1.
Authors: Sive Hazel L. , Grainger Robert M. , Harland Richard M. 
Published: May 2007

INTRODUCTIONThis article describes the creation and maintenance of tools for use in dissection and micromanipulation of embryos. All tools must be kept clean and rinsed with 70% ethanol to keep them sterile.
Housing and Feeding of Xenopus laevis
Authors: Sive Hazel L. , Grainger Robert M. , Harland Richard M. 
Published: May 2007

INTRODUCTIONGood animal husbandry is vital for maintaining a healthy frog population. This requires some effort but is generally rewarded by high-quality egg and embryo production. A healthy frog is placid, with moderately slimy skin and a nice pear shape. Jumpy frogs, frogs with dry or excessively slimy skin, bloated frogs, and frogs that look gray and thin or reddish are not healthy and should not be used for egg collection, as this would lead to further deterioration of the animals’ condition, and the resulting eggs would be generally unsuitable for experimental purposes. This article describes proper husbandry techniques for Xenopus laevis in the laboratory.
The importance of unmanaged coastal wetlands to waterbirds at coastal Ghana
Authors: Gbogbo Francis 
Published: May 2007

AbstractGhana's 550 km coastline has about 100 wetlands out of which five large ones are managed as Ramsar sites, the rest being unmanaged. Recent pollution and misuse of unmanaged wetlands have necessitated a study into their roles in the support of waterbirds. Waterbird survey on four wetlands between September 2005 and April 2006 revealed that 51 waterbird species make use of the managed compared to 44 in the unmanaged wetlands. Average numbers of waterbirds were higher on the managed wetlands (10,510 ± 4862) than on the unmanaged wetlands (1348 ± 602) (P < 0.05) but mean population density of waterbirds (n/ha) was the same for both the managed (1860 ± 310) and unmanaged (1400 ± 220) (P > 0.05) wetlands. The population densities of waterbirds belonging to guilds 1, 5 and 7 were significantly higher on the managed wetlands (P < 0.05). In contrast, guilds 2 and 4 have significantly higher population densities on the unmanaged wetlands while the population density of guild 3 was the same (P > 0.05). There was an indication that during mid and late periods of the nonbreeding season, the unmanaged wetlands altogether may support a larger number of waterbirds belonging to guilds 2, 3 and 4 than the five managed wetlands.
Abnormal puberty in paediatric Cushing's disease: relationship with adrenal androgen, sex hormone binding globulin and gonadotrophin concentrations
Authors: Dupuis C. C. , Storr H. L. , Perry L. A. , Ho J. T. F. , Ahmed L. , Ong K. K. , Dunger D. B. , Monson J. P. , Grossman A. B. , Besser G. M. , Savage M. O. 
Published: May 2007

SummaryObjective  Paediatric Cushing's disease is frequently associated with abnormal puberty. We addressed the hypothesis that prepubertal patients show excessive virilization and pubertal patients show suppression of LH and FSH secretion.Design and measurements  Serum androstenedione (A4), dehydroepiandrosterone sulphate (DHEAS), testosterone (T), and sex hormone binding globulin (SHBG) were determined at diagnosis and converted to standard deviation scores. LH, FSH concentrations were also determined. Severity of CD was assessed from the sleeping midnight cortisol concentration. Puberty was staged and excessive virilization defined as advance in pubic hair stage for breast stage or testicular volume (TV).Patients  Twenty‐seven CD patients (17 male, 10 female), median age 13·4 years (range 5·9–17·8) were studied.Results  In the CD group as a whole, A4, DHEAS, T standard deviation scores (SDS) values were normal. SHBG SDS values (n = 19) were low (median –1·93, –4·32–0·86) correlating with BMI (r = –0·49). A4, DHEAS, T, SHBG, LH and FSH did not correlate with midnight cortisol, but A4 and T SDS correlated with ACTH at 09.00 h (both r = 0·51). Thirteen patients (11 male, 2 female) had excessive virilization with increased A4 (P = 0·033), DHEAS (P = 0·008), testosterone (P = 0·033) and decreased SHBG (P = 0·004) compared with subjects without excessive virilization. Pubertal boys (TV ≥ 4 ml) (n = 7) and girls (breasts ≥ stage 2) (n = 8) had low median LH and FSH. Boys had an LH concentration of 1·2 mU/l (0·3–3·5), FSH, 0·9 mU/l (0·2–6·4) and median T SDS, –1·95 (–3·8–4·65), while girls had an LH concentration of 1 mU/l (0·3–7·4).Conclusions  Many patients had abnormal puberty and excessive virilization associated with increased adrenal androgens and decreased SHBG. Pubertal patients had low LH and FSH suggesting impaired pituitary–gonadal axis function.
The adaptation and relationship of FGF‐23 to changes in mineral metabolism in Graves’ disease
Authors: Park Se Eun , Cho Mi Ae , Kim Se Hwa , Rhee Yumie , Kang Eun Seok , Ahn Chul Woo , Cha Bong Soo , Lee Eun Jig , Kim Kyung Rae , Lee Hyun Chul , Lim Sung‐Kil 
Published: May 2007

SummaryObjective  The aim of this study was to observe the changes in bone and mineral metabolism and to confirm the regulation of fibroblast growth factor‐23 (FGF‐23) in untreated Graves’ disease.Patients and measurements  The study comprised 39 patients, with or without Graves’ disease. The Graves’ disease group was made up of 21 newly diagnosed patients, enrolled before starting treatment. Their disease was determined by biochemical and radiological means. The control group was composed of 18 people who were proven to be euthyroid without any diseases affecting bone and mineral metabolism. FGF‐23, calcium, phosphate, PTH, 25‐hydroxyvitamin D [25(OH)D] and 1,25‐dihydroxyvitamin D [1,25(OH)2D] levels and bone turnover markers were compared between these groups.Results  Serum calcium and phosphate, plasma FGF‐23 and free T4 were significantly higher in the Graves’ disease group than in the healthy control group (P < 0·05). The bone turnover markers serum osteocalcin and C‐terminal cross‐linked telopeptide of type 1 collagen (s‐CTx) were also significantly elevated in the Graves’ disease group, and had a positive correlation with free T4 levels. However, there was no significant decrease in PTH and 1,25(OH)2D in the Graves’ disease group. Plasma levels of FGF‐23 exhibited a positive correlation with serum phosphate levels and with free T4 levels (P < 0·05).Conclusions  These findings suggest that FGF‐23 is physiologically related to serum phosphate homeostasis, as indicated indirectly by the changes in bone and mineral metabolism, in untreated Graves’ disease.
Reduced maternal corticosteroid‐binding globulin and cortisol levels in pre‐eclampsia and gamete recipient pregnancies
Authors: Ho Jui T. , Lewis John G. , O’Loughlin Peter , Bagley Christopher J. , Romero Roberto , Dekker Gus A. , Torpy David J. 
Published: May 2007

SummaryObjective  To measure and contrast maternal cortisol and corticosteroid‐binding globulin (CBG) levels in pregnancies with normal outcomes, pre‐eclampsia, intrauterine growth restriction (IUGR) and in gamete recipients.Study design  Prospective study of 93 women at high risk of pre‐eclampsia, including gamete recipients (n = 22) and 33 controls. Plasma total and free cortisol and CBG were measured every 2 weeks from 16 weeks’ gestation until delivery.Results  Forty‐two per cent of the high‐risk group had complications, including pre‐eclampsia (n = 11), gestational hypertension (n = 16) and small‐for‐gestational‐age (SGA) neonates (n = 12). There were no complications in the controls. In all groups, plasma CBG concentrations increased progressively across gestation (P < 0·05), in parallel to total cortisol, but fell significantly from 36 weeks’ gestation onwards, with a corresponding rise in free cortisol concentrations. In pre‐eclampsia and gestational hypertension, plasma CBG, and total and free cortisol concentrations were lower from 36 weeks onwards (P < 0·05). In IUGR, plasma CBG concentrations were suppressed from 28 weeks’ gestation until delivery (P < 0·05), but with no significant difference in plasma total and free cortisol. Gamete recipients had significantly lower plasma CBG from 20 weeks’ gestation onwards, and plasma total and free cortisol were reduced at 24 and 32 weeks onwards, respectively.Conclusions  Maternal plasma CBG, total and free cortisol concentrations are reduced in pre‐eclampsia/gestational hypertension, and markedly reduced in gamete recipients. Low CBG may be due to reduced synthesis or enhanced inflammation‐driven degradation. Low maternal cortisol may be due to a lack of placental corticotropin‐releasing hormone or reduced maternal ACTH, driving cortisol production. Low maternal cortisol may influence the foetal hypothalamic–pituitary–adrenal axis and disease patterns later in life following complicated pregnancy.
Thyroid cysts: a new extra‐adrenal site of aldosterone synthase expression and increased aldosterone content
Authors: Greenman Yona , Trostanetsky Yana , Ben‐Shemen Sarit , Grazas Nili , Limor Rona , Osher Etty , Lewicka Sabina , Vecsei Paul , Stern Naftali 
Published: May 2007

SummaryBackground  The rapid re‐accumulation of fluid following aspiration of thyroid cystic lesions suggests that active transport of sodium and water may be involved in volume regulation of these lesions. In this study we address the possibility that aldosterone may take part in this process.Subjects and methods  Thirty‐one patients (29 women and two men), with a mean age of 52·7 ± 13·2 years (range: 27–77 years) underwent evaluation for thyroid nodules that had a sonographic cystic component. Cystic fluid obtained by FNA biopsy was sent for cytological examination and biochemical measurements. In 10 patients, material was collected for RNA extraction and determination of aldosterone synthase expression by RT‐PCR amplification.Results  All lesions were benign, cystic, colloid nodules. Cyst fluid aldosterone levels as measured by routine radioimmunoassay (RIA) were elevated above the normal plasma levels in all but five patients. Mean aldosterone levels were 27·1 ± 22·9 ng/dl (SD) (range: 5·9–117·5 ng/dl). In contrast, cyst cortisol values were in the low, low normal serum range (6·2 ± 2·9 µg/dl, range: 0·2–10·2 µg/dl). Sodium, chloride and potassium levels were 137 ± 4·7 mEq/l, 98 ± 5 mEq/l and 4·9 ± 1·4 mEq/l, respectively. Plasma aldosterone levels were normal in all patients tested. To confirm these results, 12 samples were assayed after extraction and chromatography using a highly specific antibody. Cyst aldosterone levels in this group were elevated above the normal serum range in all but one patient (mean concentration: 24·5 ± 14·6 ng/dl, range: 8·72–40·1 ng/dl). In this group, 18(OH)B levels were within the normal plasma range (12–55 ng/dl) in all but one patient (34·9 ± 17 ng/dl). Furthermore, aldosterone synthase mRNA expression was found in aspirates of four of 10 patients.Conclusions  The increased aldosterone concentration and the presence of aldosterone synthase expression suggest that aldosterone may be locally produced and secreted in thyroid tissue. The pathophysiological implications of this finding remain to be established.
Genetic analysis of the PAX8 gene in children with congenital hypothyroidism and dysgenetic or eutopic thyroid glands: identification of a novel sequence variant
Authors: Tonacchera Massimo , Banco Maria Elena , Montanelli Lucia , Di Cosmo Caterina , Agretti Patrizia , De Marco Giuseppina , Ferrarini Eleonora , Ordookhani Arash , Perri Anna , Chiovato Luca , Santini Ferruccio , Vitti Paolo , Pinchera Aldo 
Published: May 2007

SummaryObjective  To analyse the coding region of PAX8 in individuals with congenital (CH) or post neonatal hypothyroidism due to dysgenetic (TD) or eutopic thyroid glands.Design and patients  Forty‐three children with CH and TD (13 agenesis, 23 ectopia, and seven hypoplasia), one subject with post neonatal onset of hypothyroidism and thyroid ectopia, 15 children with CH and eutopic thyroid glands and six euthyroid adults with thyroid hemiagenesis were enrolled as cases, along with 120 healthy individuals as controls.Measurements  Exons 2–8 of the PAX8 were directly sequenced. HeLa and HEK293 cells were transfected with PAX8 wild‐type (PAX8‐WT), mutant PAX8, p300, thyroid transcription factor 1 (TTF‐1) and thyroglobulin promoter pGL3 (TG prom‐pGL3). Synergism of TTF‐1 with PAX8‐WT vs. mutant and activity of PAX8‐WT vs. mutant in accompaniment with p300 on TG prom‐pGL3 were also assessed. The luminescence produced by PAX8‐WT and mutant PAX8 was measured.Results  Among patients and controls only a 15‐year‐old girl with thyroid ectopia showed a heterozygous transition of cytosine to thymine at position 674 in exon 6, which changed a conserved threonine at position 225 to methionine (PAX8‐T225M). Her father and sister harboured PAX8‐T225M without abnormal thyroid phenotypes. PAX8‐T225M and PAX8‐WT similarly increased luciferase activity and had a similar synergistic effect with TTF‐1. At 500 ng p300, however, PAX8‐T225M could not significantly increase TG promoter activity when compared to PAX8‐T225M alone, while PAX8‐WT +500 ng p300 induction was significantly higher than PAX8‐WT alone (P < 0·001). Cotransfection of TTF‐1 together with PAX8‐T225M resulted in rescuing of the lack of synergism with p300.Conclusions  PAX8 mutations in congenital hypothyroidism due to dysgenetic or orthotopic thyroid glands are rare. PAX8‐T225M is probably a rare variant.
Quantitative measurement of thyroid blood flow for differentiation of painless thyroiditis from Graves’ disease
Authors: Ota Hisashi , Amino Nobuyuki , Morita Shinji , Kobayashi Kaoru , Kubota Sumihisa , Fukata Shuji , Kamiyama Naohisa , Miyauchi Akira 
Published: May 2007

SummaryObjective  Differentiation between destruction‐induced thyrotoxicosis and Graves’ thyrotoxicosis is important for selection of proper therapy. It is, however, often difficult to make this distinction without measurement of radioactive iodine uptake. We investigated the possibility that assessment of thyroid blood flow would allow differentiation between the two entities.Patients and measurements  One hundred and fourteen untreated patients with thyrotoxicosis (56 Graves’ disease, 28 painless thyroiditis, 30 subacute thyroiditis) and 25 normal controls were examined. Serum levels of freeT4 (FT4), freeT3 (FT3) and TSH were measured by chemiluminescent immunoassay, and anti‐TSH receptor antibodies (TSH‐binding inhibitory immunoglobulin, TBII) were measured by enzyme‐linked immunosorbent assay. Thyroid volume and blood flow (TBF) were measured quantitatively by ultrasonography.Results  TBF was significantly higher in Graves’ disease (mean ± 1SD: 14·9 ± 6·4%, P < 0·0001) than in painless thyroiditis (0·8 ± 0·5%), subacute thyroiditis (0·9 ± 0·7%) and in normal controls (0·8 ± 0·5%). All patients with Graves’ disease had TBF values of more than 4% and all patients with painless thyroiditis and subacute thyroiditis had TBF values less than 4%. TBF values significantly correlated with values of radioactive iodine uptake (RAIU) either at 3 h (r = 0·492, P < 0·01) or 24 h (r = 0·762, P < 0·001) within the Graves’ disease and painless thyroiditis groups. There was no relationship between TBF values and thyroid volumes or values of TBII in the Graves’ disease group. All patients with Graves’ disease had positive TBII of 15% or more. Three of 28 patients with painless thyroiditis and one of 30 patients with subacute thyroiditis had positive TBII.Conclusion  TBF was quantitatively measured by power Doppler ultrasonography and was more effective than TBII for differentiation between destruction‐induced thyrotoxicosis (painless or subacute thyroiditis) and Graves’ thyrotoxicosis. TBF values of less than 4% in untreated thyrotoxic patients are laboratory signals of destruction‐induced thyrotoxicosis and if these are determined, the radioactive iodine uptake test can be omitted for differential diagnosis of these two types of thyrotoxicosis.
Lack of electrocardiographic changes in women with polycystic ovary syndrome
Authors: Orio Francesco , Palomba Stefano , Cascella Teresa , Manguso Francesco , Vuolo Laura , Tafuri Domenico , Vigorito Carlo , Lombardi Gaetano , Liguori Vincenzo , Colao Annamaria , Giallauria Francesco 
Published: May 2007

SummaryObjective  The aim of the present study was to investigate the potential alterations in electrocardiographic (ECG) pattern in patients with polycystic ovary syndrome (PCOS).Patients  Fifty PCOS patients and 50 age‐ and body mass index‐matched healthy women were studied.Methods  We assessed hormonal and metabolic pattern, and performed ECG analysis for evaluating PQ interval, QRS duration, minimum and maximum QT interval corrected for heart rate (QTcmin and QTcmax, respectively), corrected QT dispersion (QTcd), corrected J point/T‐wave interval (JTendc), corrected JTmax interval (JTmaxc), and corrected Tmax‐end interval (Tmax‐endc).Results  QTcmin (399 ± 21 vs. 396 ± 25 ms, P = 0·51); QTc max (445 ± 25 vs. 443 ± 27 ms, P = 0·70); and QTcd (46 ± 13 vs. 47 ± 15 ms, P = 0·72); JTendc (337 ± 14 vs. 336 ± 16 ms1/2, P = 0·74); and JTmaxc (256 ± 22 vs. 258 ± 21 ms1/2, P = 0·64); Tmax‐endc (81 ± 18 vs. 78 ± 19 ms1/2, P = 0·42) were not significantly different between PCOS and healthy women.Conclusion  Despite profound differences in hormonal and metabolic pattern, our data demonstrate no significant difference in ECG pattern in PCOS compared to healthy controls.
The shrubs of the Western Ghats (South India): floristics and status
Authors: Krishnan Rani M. , Davidar Priya 
Published: May 2007

Abstract. This study examines the understorey shrub community in a wet evergreen forest of South India. The shrub community in the evergreen forests of the Western Ghats, Eastern Ghats, Sri Lanka, South‐east Asia, Neotropics, Africa and Madagascar are compared. The shrub community is richer in the Old World Tropics as compared to the Neotropics. The common families in the Old World Tropics are: Rubiaceae, Acanthaceae, Euphorbiaceae, Myrsinaceae and Annonaceae. The number of families constituting the understorey shrub community falls from forty‐six (Western Ghats) to thirteen (Central Amazon). The study concludes that the evergreen forests in the Western Ghats probably have the richest understorey shrub community.Résumé. L'étude concerne la communauté d'arbustes de sous‐bois d'une forêt sempervirente du Sud de l'lnde. Les communautés d'arbustes dans les forèts sempervirentes des Ghâts Occidentaux, des Ghâts Orientaux, du Sri Lanka, du Sud‐Est Asie, des Néotropiques, de l'Afrique et de Madagascar sont comparées. La communauté d'arbustes est plus riche dans le Vieux Monde Tropical que dans les Néotropiques. Les families communes dans le Vieux Monde Tropical sont; Rubiaceae, Acanthaceae, Euphorbiaceae, Myrsinaceae et Annonaceae. Le nombre de families constituant la communauté d'arbustes de sous‐bois varie de 46 (Ghâts Occidentaux) à 13 (Amazonie Centrale). L'étude conclue que les forêts sempervierentes des Ghàts Occidentaux ont probablement la plus riche communauté d'arbustes de sois‐bois.Mots clés. Communauté d'arbustes de sous‐bois, forêts sempervirentes, Ghâts Occidentaux, endémisme, Inde.
The vegetational history of the northern Apennines, Italy: information from three new sequences and a review of Regional vegetational change
Authors: Watson Clare S. 
Published: May 2007

Abstract. The results of pollen analysis and radiocarbon dating are presented from three northern Apennine sequences; Lago Padule in northern Tuscany and Lago Pratignano and Ospitale in the Emilia‐Romagnan Apennines. This is the first detailed pollen stratigraphic information from sites in Emilia‐Romagna and north Tuscany and extends eastwards the area from which information on vegetational history is available.The sequence from Lago Padule is one of the most complete Holocene records known from the northern Apennine region. Lago Pratignano has the deepest sequence of organic sediments (1544 cm) and the fastest rates of sediment accumulation (up to 28 cm per 100 years) providing the highest resolution record for the mid to late Holocene periods in the region. High rates of sedimentation have also occurred at Ospitale where organic sediments are 780 cm deep and began to form at approximately 5500 bp.The record from Lago Padule is compared with Holocene records from two nearby sites and a series of Regional Pollen Assemblage Zones is defined for the eastern area of the northern Apennines. The main features of vegetational change identified are: (i) a‘pioneer’phase of rapid forest development during the early Holocene followed by the establishment of an upper forest belt dominated by Abies, and a belt of mixed deciduous forest at lower altitudes; (ii) the appearance and rapid expansion of Fagus between approximately 5200 bp and 2900 bp forming a mixed Abies‐Fagus association in the upper forest belt; and (iii) the overall reduction of forest cover, and dominance of Fagus in the arboreal vegetation from around 2900 bp.The scheme of Regional PAZs for the eastern area is used as a framework for the review of pollen stratigraphic information and radiocarbon dates from other sites in the northern Apennine region. Characteristics of pollen records which can be identified in sequences from across the region are identified and the chronology of similar changes in pollen stratigraphy is examined. The scheme of four regional PAZs for the Holocene period is shown to be valid for the entire northern Apennine region. The examination of a series of pollen records shows that Fagus appeared earlier in the western than the eastern part of the region during the mid Holocene and became dominant in the northern Apennine forests post 3000 bp. The impact of anthropogenic activity and climatic change on the spread and development of Fagus are discussed. The palynological evidence which is now available from the northern Apennines is compared with information for the region shown in the‘European pollen maps’of Huntley & Birks (1983). This study provides an updated review of the representation of different tree taxa in Holocene pollen records from the northern Apennines and illustrates the role of the northern Apennines as a refugium for trees during the Wurmian Lateglacial.