Datasets:

IanTseng

/

Med-term1

like 0

to investigate the molecular mechanisms of the inhibition of nakadenosine triphosphatase nakatpase in diabetes mellitus we incubated nakatpase purified from human placenta of six healthy nondiabetic women with plasma from six IDD diabetic iddm men and six healthy controls and with different concentrations of lysophosphatidylcholine lpc we determined the enzyme activity anthroyl ouabainbinding capacity dissociation constant kd and average lifetime values tau by the static and dynamic fluorescence of anthroyl ouabain the lipid annulus of the enzyme was studied by static and dynamic fluorescence of trimethylaminophenylphenylhexatriene tmadph moreover we studied the lipid microenvironment surrounding the nakatpase purified from the placentas of six healthy women and six insulindependent diabetic women determining the percent composition of phospholipids of the lipid annulus the addition of total and proteinfree iddm plasma to normal nakatpase significantly inhibited the enzymatic activity even at the lowest concentration studied whereas the ouabainbinding capacity kd and tau were not affected by iddm plasma the fluorescence polarization and lifetime values of tmadph were significantly decreased by diabetic plasma the incubation of nakatpase with lpc caused an inhibition of the enzymatic activity without modifications of the anthroyl ouabainbinding capacity and KB the fluorescence polarization and lifetime values of tmadph were significantly decreased by mumoll lpc the study of the phospholipids surrounding nakatpase demonstrated a significant increase in the percent lpc content in iddm patients compared with controls together with a concomitant decrease in phosphatidylcholine these observations indicate that the inhibition caused by diabetic plasma on nakatpase is not dependent on a modification of the ouabainbinding site and that it seems to mimic the effect of lpc addition a link between modification of the lipid moiety of the enzyme and nakatpase inhibition might be hypothesized

31|189

insulindependent|dissociation constant

cypa the predominant cytochrome p expressed in human liver is responsible for the metabolism of endogenous CS and many drugs on the basis of pharmacokinetic studies in patients with hormonal derangements and the effects of replacement therapy it has been suggested that iodothyronines decrease cypamediated drug metabolism whereas glucocorticoids and gh enhance cypa activity the aim of the present study using well differentiated human HCs in primary culture was to examine directly whether hormonal factors regulate cypa gene expression addition of t to primary HCs resulted in a marked reduction of cypacatalyzed testosterone betahydroxylase activity and corresponding C2 of cypa protein and messenger ribonucleic acid compared to those in untreated cells conversely both dexamethasone and gh treatment substantially increased cypa mRNA none of the hormones studied consistently altered the expression of other human cytochrome p genes we conclude that iodothyronines glucocorticoids and gh act directly on human hepatocytes to regulate the expression of cypa and these effects appear to be exerted at a pretranslational level altered regulation of hepatic cypa is therefore likely to account for previous observations concerning the effects of endocrine diseases and hormonal treatments on human cytochrome pmediated drug and steroid metabolism

16|64|84|97|120

steroids|hepatocytes|hepatocytes|levels|gene expression

the possible existence of an autocrineparacrine role for srif in normal and neoplastic thyroid PF c cells has supported the use of srif analogues in the treatment of patients with medullary thyroid carcinoma mtc in this study we have investigated the expression of srif by immunohistochemistry and rtpcr and the expression of srif receptor sstr subtypes by rtpcr in a series of mtcs srif messenger rna was detected in all cases although immunoreactive cells were only identified in sstr messenger rna was present in out of the PT expression of more than sstr AT1 was detected in PT sstr the subtype that preferentially binds to the srif analogue octreotide was the AT1 most frequently detected whereas sstr was not detected in any case these results confirm the frequent expression of both srif and its receptors in mtc the presence of different combinations of sstr subtypes in a given patient may explain the variable clinical response to srif analogues and may promote the search for more selective drugs with different affinities to the various receptor subtypes

14|87|93|97|111

parafollicular|tumors|subtype|tumors|subtype

estrogen is essential for the development and maintenance of optimal bone mass in women and men and acts through activation of estrogen receptors er we have examined the pathways of estrogen action on the skeleton by seeking to localize the classical estrogen receptor er alpha to particular cells to test the hypotheses that estrogen directly influences growth SP chondrocytes and estrogen has a principal action on bone tissue via osteoblasts er alpha messenger ribonucleic acid mrna was localized by in situ hybridization in human specimens from five males yr old two females and yr old and three growing rabbits in all of the human material examined er alpha mrna was consistently identified in chondrocytes in all of the rabbit tissue studied er alpha mrna was localized in chondrocytes of the GP and the subarticular epiphyseal growth center er alpha mrna signals were readily observed in both active osteoblasts and lining cells on trabecular surfaces of all samples no clear evidence of positive staining was detectable in osteoclasts or osteocytes in either species the distribution of er alpha mrna coincided with immunolocalization of the er protein in the human specimens these data suggest a direct action of estrogen on growth plate chondrocytes that may affect longitudinal growth and subsequent F0 of the growth plate and also on OBs to affect bone formation at trabecular sites

57|130|208|216

plate|growth plate|fusion|osteoblasts

in addition to the wellknown modulation of immune and inflammatory responses the interleukin il system has been shown to be involved in the regulation of anterior pituitary hormone secretion and growth we previously demonstrated that il receptor antagonist ilra is expressed in human pituitary adenomas cultured in vitro in the present study we investigated the regulation of ilra protein by il beta uml in human somatotroph adenomas n cultured for h il beta significantly enhanced the concentration of ilra dose dependently in the somatotroph adenoma cell lysates whereas ilra concentrations remained unchanged in the culture S9 furthermore basal ilra concentrations were significantly higher in the cell lysates compared with the corresponding SC supernatants the regulation of ilra in somatotroph adenoma cells is different from human cultured monocytes in which il beta significantly stimulated ilra secretion into the SNs and no change of intracellular ilra content was observed incubation of the somatotroph adenoma cells with uml il beta did not result in a NC of gh concentrations in the SC S9 enhancement of intracellular ilra protein by il beta may represent a mechanism intrinsic to somatotroph adenoma cells to counterregulate the response to il beta on hormone secretion or cellular growth

95|111|137|162|168|169

supernatants|culture|culture supernatants|change|culture|supernatants

several constitutively activating mutations have been demonstrated in the C6 transmembrane helix of the human lh receptor hlhr in boys with gonadotropinindependent precocious puberty in the current study we examined two unrelated brazilian boys with gonadotropinindependent precocious puberty caused by two different heterozygous activating mutations of the hlhr direct sequencing of the entire exon of the hlhr revealed a heterozygous substitution of t for g at nucleotide that converts leu to arg in the third TMH of the hlhr in one affected boy his biological parents had a normal hlhr gene CS establishing the sporadic nature of this novel leuarg mutation human embryonic cells expressing hlhr mutant lr or hlhr wildtype bound cg with high affinity however cells expressing hlhrlr exhibited significantly higher basal levels of camp to fold than cells expressing the wildtype receptor indicating constitutive activation of hlhrlr basal levels of camp in hlhrlrexpressing cells were nonetheless not as great as the C2 of camp produced by hlhr wildtypeexpressing cells incubated with a saturating concentration of cg furthermore cells expressing hlhrlr were unresponsive to further stimulation by cg this finding was confirmed in the patient by lack of an increase in ST after cg stimulation these results suggest that the conformation of hlhrlr mutant represents a different activated receptor state r than the agonistoccupied wildtype receptor we also identified the previously described alaval mutation in the third intracellular loop of the lhr in the other affected africanbrazilian boy and his normal prepubertal sister suggesting the inherited form of precocious puberty in this boy we conclude that the third transmembrane helix is a potential area for activating mutations of the hlhr that cause male precocious puberty

9|75|91|154|193

sixth|transmembrane helix|sequence|levels|serum testosterone

a possible pathogenic polymorphism in the beta adrenergic receptor gene trparg has been reported to be associated with increased body weight clinical features of insulin resistance and early development of type diabetes mellitus in several populations however such findings have not been consistent among studies making the hypothesis that this genetic marker is associated with clinical features controversial to assess the effect of the genotypes on body mass index bmi we performed a metaanalysis of the data from the literature using an extension of anova for continuous measures in a total of subgroups containing subjects with n and without n the trparg variant the summary weighted mean difference in bmi was confidence interval kgm indicating that variant carriers exhibited higher bmi on the average kgm higher than normal homozygous subjects in this case there was no significant evidence against homogeneity of the effect p this is the first metaanalysis assessing quantitative phenotypes in relation to a genetic polymorphism and the results support the hypothesis that the trparg polymorphism is associated with bmi across diverse population groups suggesting that the beta adr gene locus plays a role in genetic predisposition to increased body weight in a universal manner

180

adrenergic receptor

major surgery is accompanied by extensive proteolysis of insulinlike growth factor igfbinding protein igfbp proteolysis of igfbp is generally believed to increase igf BA due to a diminished affinity of the igfbp fragments for igfs we have investigated patients undergoing elective ileoanal jpouch surgery patients were randomized to treatment with gh iuday n or placebo n from days before to days after T0 free igfi and igfii were measured by ultrafiltration of serum and igfbp proteolytic activity was determined by a irecombinant human igfbp Kd assay in the ghtreated group total igfi increased preoperatively by postoperatively total igfi decreased by PL and gh immunoassayable igfbp decreased by placebo and gh in the placebotreated group free igfi was unchanged throughout the study in the ghtreated group free igfi increased by preoperatively and remained elevated T3 operation igfbp proteolytic activity increased by after operation the relative elevations of free igfi C2 despite decreased total igfi levels could thus relate to augmented igfbp proteolysis

23|62|84|100|133|148

bioavailability|operation|degradation|placebo|after|levels

uncoupling protein ucp is a mitochondrial protein expressed in a wide range of human tissues by uncoupling respiration from atp synthesis ucp might be involved in the control of energy expenditure we have investigated ucp mRNA in human adipose tissue in eight subjects we found a positive correlation r p between subcutaneous and visceral fat depots ucp messenger rna mrna levels suggesting that ucp mrna level in subcutaneous adipose tissue is a good index of ucp mRNA in whole body adipose tissues the effect of a day verylowcalorie diet un ucp mrna level and resting VO2 was investigated in eight obese premenopausal women there was no difference in ucp mrna levels before and during the diet after days of hypocaloric diet a positive correlation was found between adipose tissue ucp mrna level and resting VO2 adjusted for lean body mass r p these results show that verylowcalorie diet unlike shortterm fasting is not associated with an induction in ucp mrna expression and that adipose tissue ucp mrna levels may be related to variations in resting energy expenditure in humans

35|76|95|134

gene expression|gene expression|metabolic rate|metabolic rate

the notion that stress activates central and peripheral pathways to inhibit the MC is well accepted but the initial processes through which this occurs have not been investigated this T0 uses a relevant nonhuman primate model to document the cyclic endocrine effects imposed by a moderate ST AS episode in the follicular phase the stress paradigm is a day inflammatoryimmunelike challenge produced by the administration of bacterial endotoxin lipopolysaccharide lps which through the PR of endogenous cytokines and other mediators induces a physiopathological response similar to a bacterial infection lps was administered iv twice daily for days starting on days of the follicular phase the stress challenge resulted in a significant lengthening of the follicular phase in all monkeys two distinct groups were observed in group n the mean se length of the follicular phase in the lpstreated cycle was significantly increased from in control cycle to days except in one monkey that had a month amenorrheic interval in group n the length of the follicular phase significantly increased but not to exceed the duration of the lps treatment vs estradiol concentrations decreased significantly after lps in group vs pgml and remained suppressed after the challenge in group estradiol C2 remained stationary throughout the day lps treatment vs compared with control values at a similar stage of the follicular phase most lh and fsh values during lps treatment were higher than controls estradiol and gonadotropin surges were delayed by lps treatment for a varying length of time according to each grp significant differences in integrated luteal progesterone concentrations characterized control cycles of groups and group group in group there were no further effects of lps on luteal progesterone during the treatment and two postlps cycles in contrast in group integrated luteal progesterone concentrations were significantly decreased in postlps cycle to cortisol significantly increased at hour T3 each morning lps injection but the amplitude of the response decreased over the day period progesterone increased significantly by hour after the first lps i.t. but remained unchanged after subsequent lps administration our data demonstrate that a day inflammatorylike episode during the FP can delay folliculogenesis and that damage to this process is intensified in individuals who already demonstrate a subtle cyclic degradation in the form of decreased progesterone secretion in the luteal phases preceding the stress episode longterm endocrine effects in the form of decreased luteal sec activity in the first poststress cycle are observed in normally cycling individuals suggesting that inadequacy of the luteal phase may represent the first stage in the damage that a stress episode can inflict upon the normal MC

12|29|46|47|73|199|305|330|348|393|430

menstrual cycle|study|shortterm|stress|release|levels|after|injection|follicular phase|secretory|menstrual cycle

subjects with poorly controlled type diabetes are both hyperglycemic and insulin resistant to determine whether short term restoration of normoglycemia improves insulin action hyperinsulinemic approximately pmoll euglycemic clamps were performed in NIDDM T3 either overnight infusion of saline or overnight infusion of insulin in amounts sufficient to maintain euglycemia throughout the night fasting glucose concentrations vs mmoll p and rates of endogenous glucose production vs mumolkgmin p were both lower T3 overnight insulin than overnight saline insulininduced stimulation of glucose uptake to vs mumolkgmin p and inhibition of free fatty acids to vs mmoll p did not differ after overnight saline and overnight insulin in contrast EGP during the final hour of the hyperinsulinemic clamps ie when glucose concentrations were the same remained higher p after overnight saline than T3 overnight insulin vs mumolkgmin thus acute restoration of euglycemia by means of an overnight insulin infusion improves hepatic and perhaps renal but not extrahepatic insulin action

31|32|70|106|129

diabetic subjects|after|after|endogenous glucose production|after

luteolysis is associated with tissue remodeling probably involving the matrix metalloproteinases mmps and their specific tissue inhibitors timps this study investigated the expression and localization of the major mmps and timps in the human corpus luteum throughout the luteal phase and T3 luteal rescue with hcg corpora lutea n were collected at hysterectomy and were dated by serial urinary lh estimation in addition corpora lutea n were collected from women who had received daily doubling doses of hcg to mimic the hormonal changes of early pregnancy mmp mmp mmp timp timp and timp were investigated by zymography reverse zymography RNA blotting and in situ hybridization there was no change in the expression of mmp timp and timp throughout the luteal phase or T3 luteal rescue little timp could be detected in the CL mmp activity peaked in the early and late luteal phase the expression and activity of mmp were maximal in the LL phase exposure to hcg during luteal rescue in vivo was associated with a reduction p in the expression and activity of mmp messenger ribonucleic acids mrnas for mmp mmp and timp were localized to the connective tissue stroma and the thecallutein cells of the CL in contrast timp mrna was localized to the granulosalutein cells and mmp mrna was expressed in scattered cells within the steroidogenic and nonsteroidogenic cell layers in conclusion during maternal recognition of pregnancy hcg prevents the normal increase in mmp in the late luteal phase mmps can function in an environment containing large amounts of timp as they have a different cellular localization

41|99|122|132|153|198

after|northern|after|corpus luteum|late luteal|corpus luteum

excess androgens are associated with a characteristic polyfollicular ovarian morphology however it is not known to what extent this problem is due to direct androgen action on follicular development vs interference with gonadotropin release at the level of the pituitary or hypothalamus to elucidate potential androgen effects on the ovary we investigated the cellular localization of androgen receptor ar mRNA mrna in rhesus monkey using in situ hybridization to investigate the regulation of ovarian ar mRNA we compared the relative abundance of ar transcripts in monkeys during follicular and luteal phases of the menstrual cycle and in monkeys treated with testosterone to assess potential PET consequences of ar expression in the primate ovary we compared ar mrna levels with indexes of follicular cell proliferation and apoptosis in serial sections from individual follicles ar mrna expression was most abundant in granulosa cells of healthy preantral and antral follicles in the primate ovary theca interna and stromal cells also expressed ar mrna but to a lesser degree than granulosa cells no significant cycle stage effects were noted in ar mrna levels however larger numbers of animals would be necessary to definitively establish a cycle stage effect ar mrna level was significantly increased in granulosa cells and was decreased in TI and stromal cells of testosteronetreated monkeys importantly granulosa cell ar mrna abundance was positively correlated with expression of the proliferationspecific antigen ki r p and negatively correlated with granulosa cell apoptosis r p in summary these data show that primate ovary ar gene expression is most abundant in granulosa cells of healthy growing follicles where its expression is upregulated by testosterone the positive correlation between granulosa ar gene expression and cell proliferation and negative correlation with PCD suggests that androgens stimulate early primate F2 development

59|75|104|207|284|291

messenger ribonucleic acid|gene expression|functional|theca interna|programmed cell death|follicle

R2 in asparaginelinked carbohydrate chains have a major impact on tsh biological properties in particular highly sialylated tsh is characterized by impaired intrinsic bioactivity and prolonged halflife the aim of the present T0 was to investigate the changes in the degree of sialylation of circulating tsh isoforms that may occur in several physiological and clinical situations bioactivity and terminal sugar residues of immunopurified tsh were studied in normal adults day and nighttime serum pools cord serum pools from normal fetuses during the third trimester fetus with primary hypothyroidism ph th week fetus with resistance to thyroid hormone rth th and rd weeks patients with ph before and during lt treatment and patients with rth before and during triiodothyrocetic acid triac treatment nighttime tsh isoforms have an increased degree of sialylation compared to daytime tsh vs p thus accounting for the lower bioactivity biologicalimmunological tsh ratio tsh bi vs p in AD ph tsh isoforms are highly sialylated p showing an impaired bioactivity p lt therapy was accompanied by a trend toward normalization of tsh biological properties tsh bi was higher p and the degree of sialylation was lower p a significant inverse correlation between tsh bi values and the degree of sialylation was observed p in normal fetuses extremely bioactive asialotsh isoforms are circulating during the rd trimester the impaired thyroid hormone action such as that occurring in hypothyroid or rth fetuses induces an early expression of alphasialyltransferase activity within thyrotropes and results in the secretion of high amounts of sialylated tsh isoforms and a hybrid tsh with peculiar terminal sugar residues and enhanced bioactivity is circulating in patients with rth tsh bi or treatment with low doses of triac can initially reduce thyroid hormone secretion in rth mainly through the secretion of tsh isoforms with changed terminal sugar residues and reduced bioactivity tsh bi in conclusion changes in the terminal sialic acid residues modulate the biological properties of circulating tsh play a relevant physiopathological role in various situations and contribute to adjust thyroidstimulating activity to temporary needs

0|32|150

variation|study|adult

expression of the nai symporter nis gene was investigated by rtpcr in a selected series of primary thyroid carcinomas PTC follicular and anaplastic fifteen follicular adenomas cold and hot adenomas were also studied five of papillary TC did not express nis messenger ribonucleic acid mrna in all but follicular cancer nis transcript was fully detected in anaplastic tissue nis mrna was only barely detected in case all of the follicular thyroid adenomas except expressed the nis gene in contrast all tumors studied excluding the anaplastic histotype fully expressed thyroglobulin and thyroid peroxidase mrna transcripts in patients a lower expression to fold of nis mrna was found in metastasis by dot blot analysis compared with those in both normal and primary neoplastic thyroid tissue four of differentiated thyroid cancer patients selected for the presence of metastases with negative posttherapy i TBS showed the lack of nis gene expression in their primary cancer this defect at least in these cases is a somatic and intrinsic lesion of the primary cancer cells and is not due to a dedifferentiation process in the metastatic tissue the early detection of the loss of nis mRNA in the primary CA therefore may provide useful information for the management of DTC patients

19|36|139|189|193|203

papillary|thyroid cancer|total body scan|gene expression|cancer|differentiated thyroid cancer

activating mutations of the extracellular calcium caesensing receptor car gene mostly in its extracellular domain can cause both familial and sporadic HP we report a japanese family with severe hypoparathyroidism with pretreatment SS calcium ca levels of mgdl the proband presented with a seizure at days of age her older brother and mother who had also experienced seizures and tetany respectively likewise had HP a heterozygous missense mutation substituting a cysteine for the phenylalanine normally present at codon fc was identified in the cars fifth transmembrane domain and was shown to cosegregate with the disease the mutation was absent in dna from control subjects analysis of the functional properties of the mutant receptor was carried out in transiently transfected hek cells loaded with fura by assessing caeevoked increases in the cytosolic calcium concentration cai there was a leftward shift in the concentrationresponse curve for the mutant receptor ec effective concentration of cae producing half of the maximal cai response vs mmoll for the wildtype receptor hek cells cotransfected with both the wildtype and mutant cars to mimic the heterozygous state in affected family members showed an ec mmoll similar to that of the mutant car alone thus we confirm that a gain of CF mutation in the fifth TM domain of the car causes severe familial hypoparathyroidism by rendering the receptor more sensitive than normal to activation by cae some patients in the family do not experience seizures despite their severe hypocalcemia and this condition needs to be differentiated from other causes of hypoparathyroidism

21|32|63|203|208

hypoparathyroidism|serum|hypoparathyroidism|function|transmembrane

the WT1 gene wt which is deleted in some wilms tumors encodes a zinc finger transcription factor we studied wt messenger ribonucleic acid mrna in human term placenta and CTs differentiating into syncytiotrophoblasts in vitro by rtpcr the results suggest that wt mrna is expressed in the trophoblasts in a cellspecific fashion wt mrna expression has been observed to decline remarkably in trophoblast cells after h when these cells are morphologically differentiated into multinucleated syncytiotrophoblasts as it is well known that camp as a second messenger plays a significant role in cellular proliferation and differentiation of placental cells we examined the effect of bromocamp on wt mrna expression in undifferentiated cytotrophoblasts and differentiated syncytiotrophoblasts we observed that camp enhanced wt mrna expression in cytotrophoblasts but remained ineffective in altering wt mrna in syncytiotrophoblasts in summary the results of this investigation demonstrate that the wt gene is developmentally regulated during trophoblast differentiation an involvement of the campmediated system in regulating the wt gene in the trophoblast is suggested

1|29

wilms tumor suppressor|cytotrophoblasts

increased serum insulinlike growth factor igfbinding protein igfbp proteolytic activity igfbppa has been demonstrated in a number of clinical states of insulin resistance including severe illness after surgery and in noninsulindependent diabetes mellitus in the present study we assessed the role of insulin sensitivity in expression of igfbppa in SS in patients studied a significant increase in igfbppa p was demonstrated after colorectal surgery eight patients receiving an oral glucose load before surgery demonstrated a significant greater relative increase in igfbppa compared with patients not receiving glucose vs respectively p both CG had reduced insulin sensitivity after surgery p n as determined by hyperinsulinemic normoglycemic clamps however the group not receiving glucose displayed less insulin sensitivity than the oral glucose load group p multiple regression analysis demonstrated that the relative changes in igfbppa and c peptide C2 were inversely correlated p suggesting that increased igfbppa presumably increasing igf BA may be associated with decreased insulin demands interestingly insulin infusion during the h hyperinsulinemic normoglycemic clamp performed h after surgery postop resulted in a further increase in igfbppa in both CG p whereas no significant responses could be demonstrated during the preop clamp the expression of increased igfbppa was accompanied by conversion of endogenous intact kda igfbp into its kda fragmented form as determined by western immunoblotting and this conversion was virtually complete after the h postop clamp in patients displaying marked increases in igfbppa characterization of the igfbppa demonstrated that it was specific for igfbp as no degradation of igfbp and was detected and the use of various protease inhibitors demonstrated that serine proteases and possibly MMPs contribute to the increased igfbppa level after surgery we propose that igf bioavailability may be increased by the induction of igfbppa in insulinresistant subjects and that insulin regulates igfbppa in this state

49|91|136|148|179|266

serum|groups|levels|bioavailability|groups|matrix metalloproteinases

we investigated the p status and the ability of exogenous wildtype wt p to affect chemosensitivity in three anaplastic TC cell lines bht sw and kat all three cell lines had nonfunctional p treatment with mitomycin c or adriamycin did not result in accumulation of p or induction of pwafcip or mdm and did not cause rb dephosphorylation bht and kat cells had mutant p sw cells were functionally mutant because of marked downregulation of wt p messenger ribonucleic acid representing a novel mechanism of p dysfunction infection with a pexpressing adenovirus adp induced high C2 of p and mdm proteins in bht cells induction of p and mdm was evident h T3 infection in kat cells induction of p and mdm was observed day after infection and continued to increase over the ensuing h sw cells demonstrated intermediate kinetics sensitivity to the cytotoxic effect of adp paralleled the kinetics of pmdm induction bht cells were most sensitive to killing by adp with an ic of less than multiplicity of infection sw cells were intermediate in sensitivity kat cells were resistant all three cell lines became more CS to adriamycin after wt p expression with a fold decrease in ic values the latter observation may make a combination of wt p and chemotherapeutic drugs an attractive modality for treating anaplastic thyroid CA

19|95|112|187|221

thyroid carcinoma|levels|after|sensitive|cancer

antioxidant defenses play a critical role in the regulation of programmed cell death even when death is induced by nonoxidative stimuli during spermatogenesis most of the testicular germ cells degenerate by an AI process that is under hormonal control however the exact mechanisms by which hormonal signals are transduced within the cells to direct their life and whether other effectors of the apoptotic pathway for example antioxidants take part in the control of human germ cell survival are not known in the present T0 testosterone and nacetyllcysteine nac which is an antioxidant an inhibitor of apoptosis in several systems and a survival factor in human semen were found to suppress programmed cell death in human testicular germ cells in vitro the samples came from adult men undergoing ORX for prostate cancer germ cell death was induced by incubating segments of ST under SF culture conditions this apoptosis detected by southern blot analysis of dna fragmentation by dna labeling in situ and by morphological analysis under the electron microscope was significantly inhibited by testosterone at concentrations of and moll nac concentrations of and mmoll suppressed germ cell death in a dosedependent manner this inhibition was ERP during and h of incubation apoptotic cells were identified mainly as spermatocytes and early spermatids PCD was also demonstrated in late spermatids we conclude that nac which is an antioxidant plays an important role in germ cell survival in the human seminiferous tubules in vitro we also suggest nac as a possible new therapeutic factor for some men with idiopathic oligospermia

32|83|127|140|142|194|210

apoptotic|study|orchidectomy|seminiferous tubules|serumfree|effective|programmed cell death

in cirrhosis as in other conditions of protein catabolism there is a state of acquired gh resistance as defined by high circulating gh levels with low insulinlike growth factor i levels however patients with endstage liver failure respond to supraphysiological doses of gh with an increase in circulating insulinlike growth factor i levels the present study represents a detailed analysis of gh receptor ghr expression in cirrhotic liver from patients with endstage liver disease specific binding of labeled gh was identified in all cirrhotic livers studied the binding affinity for the ghr was similar in cirrhotic and normal livers but the number of binding CS per mg protein of liver membrane was variable in both normal and cirrhotic liver although it were generally lower in cirrhotic liver ghr expression was identified in cirrhotic CL by RNA blotting rtpcr and ribonuclease protection CA on RNA blotting a single transcript of kb was identified in normal and cirrhotic tissues rtpcr identified expression of both FL ghr and a truncated form of the ghr this was confirmed by ribonuclease protection CA in situ hybridization and immunohistochemistry confirmed the expression of ghr in regenerating HCs and isolated cells in fibrous tissue in conclusion the low level of ghr in cirrhotic CL may contribute to the acquired gh resistance found in cirrhotic patients the reduced expression of both fulllength and truncated ghr is compatible with the low level of ghbinding protein found in cirrhosis as this truncated receptor has previously been reported to generate large amounts of ghbinding protein and the demonstration of gh IB to cirrhotic liver explains why these patients with gh resistance may still respond to supraphysiological doses of gh

104|133|135|141|143|162|177|190|206|259

sites|liver|northern|assay|northern|fulllength|assay|hepatocytes|liver|binding

there is increasing evidence that crh which is the principal neuroregulator of the hypothalamicpituitaryadrenocortical axis is also involved in the mechanism of human labor the human myometrium has been shown to express several high affinity crh receptors although the identities of the crh receptor subtypes have yet to be identified to investigate further the expression of the crh receptor in human myometrium we used rtpcr fluorescent in situ hybridization and immunofluorescence to identify and localize the four subtypes alpha beta alpha and the variant c of the crh receptor interestingly the crh receptor subtypes in myometrium exhibit differential expression patterns in human pregnant myometrium at term all four receptorsubtypes were expressed whereas only the alpha and betareceptor subtypes were found in the nonpregnant myometrium this would suggest that crh acting via different receptor subtypes is able to exert different actions on the myometrium in the pregnant state compared to the nonpregnant state furthermore in the Pr human uterus crh receptors were localized in both smooth muscle and fibroblasts these findings suggest that crh receptor expression plays an important modulatory role in myometrial and possibly in C2 CF

155|185|186

pregnant|cervical|function

prl is synthesized by decidualized endometrial stromal cells from the midsecretory phase in a nonconception cycle and throughout pregnancy the exact role of prl in the human endometrium remains to be elucidated however the pattern of expression supports a role for prl during implantation and placentation this study investigated the site and DP of expression of prl receptors in the nonpregnant human endometrium in situ hybridization and immunohistochemistry localized expression of the receptor in the GE and a subset of stromal cells of the endometrium as judged by the intensity of staining expression of the receptor was dramatically upregulated during the secretory phase expression of the prl receptor gene in the endometrium from the sec phase of the menstrual cycle was confirmed by ribonuclease RPA using micrograms total ribonucleic acid phosphorylation of janus kinase jak stat signal transducer and activator of transcription and stat proteins in response to prl was investigated to establish the signaling pathway of prl in the human endometrium endometrial tissue was collected during the secretory phase of the menstrual cycle and incubated in the presence of ngml human prl for and min jak phosphorylation was induced by prl at min whereas stat and stat phosphorylation was apparent min T3 stimulation with prl immunohistochemistry localized the jakstat proteins in the glandular epithelial cells and a subset of stromal cells as was observed for the prl receptor sec phase stromal and glandular cells cultured separately and in the presence or absence of ngml prl confirmed the prlinduced phosphorylation of jakstat proteins at least in the glandular compartment these studies demonstrate an upregulation of expression of functional prl receptors during the secretory phase of the menstrual cycle further decidual prl through a paracrine mechanism may influence glandular epithelial functionsecretions and direct gene transcription through the jakstat pathway the target genes activated by prl in the glandular epithelium of the nonpregnant human endometrium remain to be elucidated

52|75|114|124|202|229

pattern|glandular epithelium|secretory|protection assay|after|secretory

a vietnamese girl with LS has been treated with recombinant human insulinlike growth factori for yr from age yr her height sd score increased from to without acceleration of BA isolated breast development PD despite pubertal suppression with luteinizing hormonereleasing hormone analogue which was stopped after yr because of growth deceleration facial coarsening was documented with serial photographs sequencing and in vitro analysis identified a homozygous base pair substitution in exon of the probands gh receptor ghr which changed amino acid from proline to glutamine pq and disrupted gh binding both the pqmutated human ghr and wildtype wt hghr were transiently expressed in cos cells as demonstrated by western blotting but the pqtransfected cells did not bind ihgh similarly fdcp cells transfected with wthghr bound ihgh with high affinity and proliferated in response to gh whereas the pq hghr cells did neither in chok cells cotransfected with wthghr and the egr promotor linked to a luciferase reporter gene gh evoked a fold increase in luciferase activity but there was no response in the cells carrying the pq hghr mutation from examination of the crystal structure of the ghr we suggest that the pq mutation disrupts the interdomain link between the extracellular domains of the ghr causing a conformational change that results in disruption of the gh IB site

4|29|33|216

laron syndrome|bone age|progressed|binding

the diagnosis of gh deficiency ghd in the elderly is based at present on the peak gh concentration during a stimulation test we have now evaluated the performance of urinary gh ugh urinary insulinlike growth factor i uigfi and urinary igfbinding protein uigfbp in the diagnosis of ghd in this group twenty ghd elderly patients with a history of pituitary disease and a peak gh response to Arg stimulation of less than ngml men and women age yr and controls men and women age yr were studied gh secretion was assessed by h profile and expressed as the area under the curve aucgh serum s igfi and sigfbp were measured in a single morning fasted sample urinary gh uigfi and uigfbp were measured in a h urine sample collected over the same interval as the gh profile and results were expressed as total amount excreted in h tugh nanograms tuigfi nanograms tuigfbp micrograms data are presented as the mean sd except for aucgh tugh and tuigfbp which are presented as the geometric mean tolerance factor aucgh sigfi and sigfbp were significantly lower in ghd subjects than in controls total ugh was lower in ghd subjects but tuigfi and tuigfbp excretion were not different in the two groups aucgh provided the best separation between ghd and control subjects whereas there was substantial overlap for sigfi sigfbp and tugh in both groups sigfi was correlated to sigfbp ghd r controls r both p whereas tuigfi was not correlated to tuigfbp in either group moreover tuigfi and tuigfbp were not related to their respective serum concentrations in either group total ugh was correlated with aucgh only in controls r p these data demonstrate that urinary gh and urinary and serum igfi and igfbp are not suitable diagnostic markers for ghd in elderly subjects

67

arginine

previous studies have suggested that metformin is clinically useful in the treatment of polycystic ovary syndrome pcos the aim of this study was to evaluate whether the improvement of ovarian function achieved by metformin therapy is associated with changes in leptin concentrations twentysix obese women with pcos were treated with mg MET x daily for months and women continued the therapy for months a significant decrease in the serum leptin level was observed after months of treatment in the whole T0 group ngml vs ngml p in the women treated for months the mean SS leptin concentration decreased after months ngml vs ngml p but slightly increased after months of treatment ngml not significant these results indicate that insulin sensitizing therapy with metformin decreases the leptin concentrations in obese pcos women

51|80|94

metformin|study|serum

either spontaneous or pharmacological stimulated gh secretion is reduced with advanced age this observation is an added difficulty for the PSA diagnosis of GHD in adults furthermore the combined administration of saturating doses of ghreleasing hormone ghrh plus ghreleasing hexapeptide ghrp is nowadays the most effective ghreleasing CS- tested in a variety of settings related to altered somatotroph function to understand whether the gh discharge elicited by the combined CS- declines with age normal subjects of both sexes divided into age CG adults yr aged yr and very old yr subjects were studied they were administered iv as bolus and in combination micrograms ghrh plus micrograms ghrp in the three groups the combined administration of ghrh plus ghrp elicited a gh area under the curve micrograml per min of and for adults aged and very old subjects respectively NS differences the mean gh peak was microgramsl for adults microgramsl for aged subjects and for very old subjects NS differences individually examined there were no nonresponders to the combined CS- and all subjects independently of age showed a gh peak over microgramsl the lowest peak was microgramsl and the highest peak was microgramsl in conclusion the ghrh plus ghrpinduced gh release is well preserved in aged and very old subjects which suggests that the gh secretory capability of the combined test is not reduced by age this combined test may be useful for the diagnosis of ghdeficient states in adults

20|23|47|69|81|138|157|168

biochemical|gh deficiency|stimulus|stimulus|groups|nonsignificant|nonsignificant|stimulus

aldosterone suppression by dexamethasone and high hydroxycortisol and oxocortisol levels are used to differentiate glucocorticoidremediable aldosteronism gra from other forms of primary aldosteronism these methods are time consuming expensive and impractical for C1 studies moreover diagnosis of gra requires a confirmatory genetic test we evaluated patients with primary aldosteronism referred to our centers by the use of a long pcr technique to reveal the chimeric gene of gra in of patients the response of aldosterone to dexamethasone mgday for days was also assessed none of our patients including pairs of siblings was positive for the chimeric gene the results of long pcr were confirmed by southern blotting despite a negative genetic test patients with aldosteroneproducing adenoma and with idiopathic hyperaldosteronism had plasma aldosterone suppressed by dexamethasone ie or ngdl of patients were identified as having aldosteroneproducing adenoma and as having idiopathic hyperaldosteronism in our experience the long pcr technique is a reliable and simple test to at least exclude gra in patients with primary aldosteronism a short term dexamethasone suppression test of aldosterone can be misleading in identifying gra the prevalence of gra in primary aldosteronism remains to be established

32

large

many men with idiopathic OP have reduced circulating insulinlike growth factori igfi levels the major source of circulating igfi is ghmediated production by the liver the known anabolic effects of gh on the skeleton raised the possibility of gh deficiency in these men we sought to test this hypothesis in this T0 fourteen men mean age yr range with idiopathic OP were studied mean lumbar spine bone mineral density bmd was gcm t score FA neck bmd was gcm t score distal radius bmd was gcm t score eleven of had frank reductions in serum igfi levels compared with age and sexmatched values sd vs sd gh secretion was stimulated by iv arginine infusion g over min followed h later by oral ldopa mg serum gh was measured at time t and min all patients responded to at least one stimulus with the majority n responding to both five patients responded either to arginine or to ldopa but not to both baseline gh for the entire group was ngml sem peak gh following Arg t min was ngml a fold rise peak gh following ldopa t min was ngml a fold rise no difference in maximal secretion was observed between those with low or normal igfi levels neither igfi nor igf binding protein concentrations changed significantly during the short period of gh stimulation these data suggest that men with OP and reduced igfi levels do not appear to have a deficiency in the gh axis other hormonal or local factors may be important in regulating igfi expression deficiencies of igfi production at skeletal CS may be important in the pathogenesis of this syndrome

4|51|60|74|173|229|263

osteoporosis|study|osteoporosis|femoral|arginine|osteoporosis|sites

serum prostatespecific antigen psa is produced in several female tissues and appears to be upregulated by androgens we have studied serum psa concentrations in women with different forms of hyperandrogenism focusing on the influence of changes in ovarian and adrenal function on these concentrations thirtyseven hirsute women were studied in the follicular phase of the MC basal and acthstimulated plasma samples were obtained and sampling was repeated gonadal stimulation and gonadal suppression days T3 receiving a single im mg dose of triptorelin eleven nonhyperandrogenic women served as controls hirsute women had increased psa levels compared to controls when considering the source of the hyperandrogenism ovarian patients those with increased SS androgen levels that normalized during gonadal suppression and adrenal patients those with increased androgen C2 that remained elevated during gonadal suppression presented increased psa values whereas hirsute patients without hyperandrogenemia had normal psa levels psa levels did not change during ovarian or adrenal stimulation or during gonadal suppression with respect to initial values basal psa C2 showed significant correlations with basal TT r p free androgen index r p sex hormonebinding AHG r p dehydroepiandrosterone sulfate r p hydroxyprogesterone r p and age r p when patients and controls were considered as a whole in conclusion basal psa C2 are increased in hirsute patients and correlate with the degree of HA when patients and controls are considered as a whole the adrenal and the ovary do not appear to be the source of psa suggesting that hyperandrogenism induces psa secretion in tissues other than the adrenal and the ovary

55|73|109|124|165|171|181|208|220

menstrual cycle|after|serum|levels|levels|total testosterone|globulin|levels|hyperandrogenism

twentyone hydroxylase pc is a key enzyme essential for normal zona glomerulosa and fasciculata function recently hydroxylase deficiency has been implicated in the pathogenesis of adrenocortical tumors therefore we investigated the mutational spectrum of the cypb gene and the messenger rna expression of pc in six aldosteroneproducing adenomas seven cortisolproducing adenomas two nonfunctional incidentally detected adenomas and four adrenal carcinomas dna from leukocytes and tumors was amplified by pcr using primers specific for the cypb gene the exons intron intron all other exonintron junctions and bp of the promoter region of cypb were automatically sequenced polya rna was extracted from tumor tissue dot blotted on a nylon membrane and hybridized with plabeled p sidechain cleavage p CYP17 and pc complementary dna probes we detected heterozygous germline mutations exon val leu in two patients one with a cortisolproducing adenoma and the other with an androgensecreting adrenocortical carcinoma a somatic heterozygous microdeletion was found in exon of one aldosteroneproducing adenoma the pc gene expression correlated with the clinical phenotype of the tumor with low pc messenger rna expression in nonfunctional adenomas compared with high pc expression in aldosterone and cortisolproducing adenomas and respectively vs normal adrenals in conclusion the prevalence of heterozygous germline mutations in the cypb gene was higher in patients with ACT confidence interval than in the GA european population confidence interval but this difference is questionable because of the low number of subjects in our series the pathophysiological significance of this finding in the presence of one normal cypb gene seems to be low suggesting that hydroxylase deficiency is not a major predisposing factor for adrenal tumor formation

116|211|217

alphahydroxylase|adrenocortical tumors|general

SS paraoxonase is a glycoprotein which binds to highdensity lipoproteins hdl and may prevent oxidation of ldl by hydrolyzing LOOH two polymorphisms identified in the paraoxonase gene metleu and glnarg have been associated with cardiovascular disease oxidative lowdensity lipoprotein ldl is also toxic to retinal capillary EC cells and pericytes so that mildly modified ldl may contribute to the development of diabetic retinopathy to investigate the potential significance of these polymorphisms in the pathogenesis of DR in iddm patients with DR and controls without DR were investigated in the current project the allelic frequency of leucine l was significantly higher in the group with retinopathy than without retinopathy vs p the TT ll was strongly associated with the OD of diabetic retinopathy p but a similar association was not found with glnarg leucine is a risk factor for DR

0|19|46|75|80|84|111|118|138

serum|lipid peroxides|endothelial|diabetic retinopathy|diabetic retinopathy|diabetic retinopathy|genotype|development|diabetic retinopathy

connective tissue growth factor ctgf is a cysteinerich peptide involved in human atherosclerosis and fibrotic disorders such as scleroderma ctgf has considerable nterminal sequence similarity with the insulinlike growth factor binding proteins igfbps including preservation of cysteines and has been postulated to be a member of the igfbp superfamily indeed recent studies have shown that BV generated ctgf a secreted kda protein binds igfs in a specific manner leading to the provisional renaming of ctgf as igfbp or igfbprp with immunoprecipitation and immunoblotting using polyclonal antiigfbprp antibody generated against recombinant human igfbprpbac igfbprp can be identified in the serumfree conditioned media of hst human breast cancer cells as well as in various human biological fluids such as normal sera pregnancy sera and cerebrospinal amniotic follicular and peritoneal fluids glycosylation studies with endoglycosidase f reveal that endogenous human igfbprp is a secreted glycosylated approximately kda protein with kda of nlinked sugars and a kda core there are and kda proteins that appear to be igfbprp degradation products in hst human breast CA cells transforming growth factor tgfbeta a potent growth inhibitor for these cells upregulates igfbprp mrna and protein C2 expression of hst igfbprp is significantly increased by tgfbeta treatment in a dosedependent manner with and fold increases in mrna and protein levels respectively at a tgfbeta concentration of ngml our studies indicate that igfbprp appears to be an important endocrine factor and one of the critical downstream effectors of the critical downstream effectors of tgfbeta similar to the role of igfbp in tgfbetainduced growth inhibition in human breast cancer cells

55|170|188

baculovirus|cancer|levels

the dax gene encodes an orphan nuclear hormone receptor essential for normal fetal development of the adrenal cortex recently dax has been shown to act as a transcriptional repressor of StAR gene expression star suppressing steroidogenesis we therefore investigated the expression of dax in a variety of adrenocortical tumors and compared the results with star mrna expression we found low or absent dax expression in aldosteroneproducing adenomas n normal adrenals and in aldosteroneproducing adrenocortical carcinomas n and cortisolproducing adenomas showed intermediate dax expression n as did nonaldosteroneproducing carcinomas and high dax expression was present in nonfunctional adenomas n in contrast to dax star mrna expression did not show significant variations between groups we did not detect the expected negative correlation between dax and star in adrenocortical PT these data suggest that high dax expression in ACT is associated with a nonfunctional phenotype whereas low dax expression favors mineralocorticoid secretion these effects on steroidogenesis are mediated by mechanisms other than repression of star gene expression our results indicate that dax may be one of the AF influencing the steroid biosynthesis of adrenocortical neoplasms

30|126|135|174

steroidogenic acute regulatory protein|tumors|adrenocortical tumors|factors

in a previous T0 we have isolated and characterized cdna encoding a novel short form of heparinbinding egflike growth factor sf hbegf loukianov et al in the present work we have found that cdna for sf hbegf and for fulllength hbegf are each represented by two variants which we refer to as l and p forms the l form is the previously known form of hbegf cdna and encodes a leucine in position the p form described in this report encodes a proline in codon the lp substitution is predicted to cause a significant alteration in the proregion structure of sf hbegf and hbegf

3

study

the abia gene encodes an abortive bacteriophage infection mechanism that can protect lactococcus species from infection by a variety of bacteriophages including three unrelated phage species five heptad leucine repeats suggestive of a leucine zipper motif were identified between residues and in the predicted CAA sequence of the abia protein the biological role of residues in the repeats was investigated by incorporating CAA substitutions via sitedirected mutagenesis each mutant was tested for phage resistance against three phages phi sk and c belonging to species p and c respectively the five residues that comprise the heptad repeats were designated l l a l and l three single conservative mutations of leucine to valine in positions l l and l and a double mutation of two leucines l and l to valines did not affect abia activity on any phages tested nonconservative single substitutions of charged amino acids for three of the leucines l l and l virtually eliminated abia activity on all phages tested substitution of the alanine residue in the third repeat a with a charged residue did not affect abia activity replacement of l with an alanine elimination phage resistance against phi but partial resistance to sk and c remained two single proline substitutions for leucines l and l virtually eliminated abia activity against all phages indicating that the predicted alphahelical structure of this region is important mutations in an adjacent region of basic amino acids had various effects on phage resistance suggesting that these basic residues are also important for abia activity this directed mutagenesis analysis of abia indicated that the leucine repeat structure is essential for conferring phage resistance against three species of lactococcal bacteriophages

44|62

amino acid|amino acid

rapid progress in sequencing of human and other genomes allows highresolution analysis of their gene content on the basis of comparison between species we have used a combined computer and PSA RPA to characterize kb of human genomic sequence from q and discovered a new exon gene termed nipsnap located between the neurofibromatosis type and the pk genes the nipsnap gene spans kb of genomic CS and shows to large introns in the region all exonintron junctions contain the gtag consensus splice site the putative promoter of the nipsnap gene is tataless and resides in a gcrich island characteristic of housekeeping genes the nipsnap mrna is kb is expressed ubiquitously at variable levels with the highest expression in liver is terminated by an uncommon attaaa polyadenylation site and is capable of encoding a AA protein this nipsnap protein has a strong CS similarity limited to the CE portion of a hypothetical protein acc p from chromosome iii of c elegans in which the other portions resemble a nitrophenylphosphatase domain and nonneuronal snaplike protein thus the nipsnap gene is a member of an evolutionarily well conserved novel gene family with two members in human and mouse that have now been characterized and one member in c elegans the second human gene nipsnap is localized in the vicinity of marker ds on chromosome although the function of the nipsnap protein family is unknown clues about its role may reside in the coexpression of the c elegans orthologue within an operon encoding protein motifs known to be involved in vesicular transport

30|31|65|133|141|146

biochemical|approach|sequence|aminoacid|sequence|central

the kda antigen of m tuberculosis which is an alanine dehydrogenase is a speciesspecific antigen that is potentially useful for CS identification large quantities of the purified protein are required for immunological as well as for detailed biochemical and structural characterization the aladh gene was cloned by pcr from hrv virulent and hra partially attenuated strains of m tuberculosis and their dna CS was determined a hostvector system suitable for the production of sufficient quantities of the recombinant aladh antigen was developed the aladh gene was expressed under the control of strong transcriptional bacteriophage plpr and translational atpe signals highlevel expression of soluble aladh was obtained using the recombinant e coli k strain cag pmsk which is deficient in lon protease and the heatshock response a simple twostep procedure for the rapid purification of the recombinant protein was developed the protein was purified to near homogeneity and the purified aladh showed a specific enzyme activity comparable to the native protein isolated from m tuberculosis in addition the product showed an expected CAA CS and reacted strongly to the kda aladhspecific mab hbt furthermore the epitope of the mab hbt was mapped to a AA region contrary to the published results we show that the aladh and the pnt pyridine nucleotide transhydrogenase of m tuberculosis do not share common epitopes reacting to the speciesspecific mab hbt the availability of highly purified aladh should now enable a detailed biochemical and structural characterization of this important enzyme of m tuberculosis

20|62|171|172|193

strain|sequence|amino acid|sequence|aminoacid

we have cloned and sequenced the pencoding cdna of green swordtail x helleri and southern platyfish x maculatus these two fish species are often used to produce hybrids that develop melanomas after genetic crossing computer translation of derived cdna sequences revealed that p polypeptides from these two species are virtually identical exhibiting only two conservative amino acid substitutions tp mrna expression was detected in virtually all tissues tested comparison of these fish p polypeptide sequences with those of other vertebrates including other fishes amphibians and mammals revealed that conservation is especially high in several previously defined protein domains in addition sequencing of the tp genomic region of x maculatus reveals similarity to the human tp locus in overall organization knowledge of the xiphophorus tp sequences will allow assessment of mutational alterations within PT generated from numerous fish genetic crosses

132

tumors

the reverse transcription polymerase chain reaction rtpcr with primers TPS for each of the exons of the human complement regulatory protein membrane cofactor protein mcpcd has been utilized to determine mcp mrna transcript expression in IP blood mononuclear cells pbmc an additional transcript of a larger size than predicted was consistently detected in reactions with a sense primer for exon that encodes the first alternatively spliced serinethreoninerich region sta together with an antisense exon primer rtpcr with primers for other exons both and of exon further showed that these mcp transcripts contain additional sequences immediately both and to the exon encoded sequence comparison of genomic dna with cdna by pcr in combination with CS analysis demonstrated the presence of the complete invariant sequences of both introns adjacent to exon ie intron bp and intron bp rtpcr using primers specific for the intron sequence together with southern and RNA blotting using an intron specific probe confirmed retention of this intron within a novel kb mrna transcript in human pbmc due to the presence of a stop codon within intron translation would result in a novel truncated mcp isoform mcpi containing the four invariant short consensus repeat scr regions and a unique cterminal amino acid transmembrane and cytoplasmic tail region that may promote endoplasmic reticulum PR

9|35|113|147|213

specific|peripheral|sequence|northern|retention

we had previously reported the purification and partial characterisation of four distinct odorantbinding proteins from male mouse nasal epithelium one of these named obpi appeared to be a heterodimer whose subunits ia and ib showed significant similarity in their nterminal amino acid sequences with hamster aphrodisin in this paper we report the complete CAA sequences of these two polypeptide chains as deduced from nucleotide sequences of their relative cdna these data confirm the high similarity of both proteins with hamster aphrodisin a comparison with the sequences of other known obps indicate that they are more closely related to members of class i including bovine obp rat obpi and pig obpi a putative odorantbinding site is indicated by the presence of amino acid residues conserved with respect to the bovine protein whose threedimensional structure has been recently resolved insitu hybridisation has revealed identical expression patterns for the two proteins further supporting the heterodimeric structure of these proteins in the nasal mucus

53

amino acid

we have shown that escherichia coli pcnb mutants are lysogenized by bacteriophage lambda with lower efficiency as compared to the pcnb strains our genetic analysis revealed that expression of the lambda cii gene is decreased in the pcnb mutants however using various lacz fusions we demonstrated that neither activities of pl and pr promoters nor transcription termination at tr were significantly impaired in the pcnb host on the other hand we found that oop rna an antisense rna for cii expression is involved in this regulation primer protection experiments revealed that oop rna was polyadenylated and that this polyadenylation was impaired in the pcnb mutant we found that the oop rna was more abundant in the pcnb mutant than in the pcnb strain furthermore we showed that activity of the po promoter was not stimulated in the pcnb mutant such findings indicated that Kd of oop rna in the pcnb strain was slower because of inefficient polyadenylation which could lead to more effective inhibition of cii expression by the antisense oop rna resulting in less efficient lysogenization of the host the oop rna was found previously to play a role in phage lambda development only under conditions of overproduction of this transcript here we demonstrate for the first time the physiological function of oop rna in lambda OD confirming that this short transcript plays an important role in the negative regulation of cii gene expression during lambda infection moreover polyadenylation of oop rna is one of very few known examples of TPS rna polyadenylation by pap i in prokaryotic cells and its role in gene expression regulation

143|217|251

degradation|development|specific

a p cloned insert of about kilobases kb was isolated containing four members of the human growth hormonechorionic somatomammotropin ghcs gene family and the thyroid HR interacting protein trip gene the presence of the cslike csa ghvariant and most downstream csb gene was confirmed by dna blotting and sequence analysis the trip gene was detected kb downstream of the csb gene and in the reverse transcriptional orientation to all the ghcs genes the trip gene is highly homologous to the sug gene in yeast and is evolutionarily conserved among several species based on the common location of the gh and trip or homologue genes on the same chromosome in the human pig and rat genomes we suggest that these loci are physically linked previously it was reported that a musclespecific sodium channel scna gene is located immediately UP of the pituitary growth hormone ghn gene and is linked to the gh gene locus in both humans and rats this suggests a further linkage between the scna gh and trip loci also deoxyribonuclease hypersensitive sites have been reported in and around these loci and were associated with an important locus control region for the ghcs genes unlike the ghcs genes we show using reverse transcriptasepolymerase chain reaction that the trip gene is expressed ubiquitously and through rna blotting as a kb transcript this implies an open and active chromatin structure the possible effect of this structure on the adjacent human ghcs gene locus is discussed

25|137

hormone receptor|upstream

an improved method for gene replacement in pseudomonas aeruginosa was developed the method employs several new gene replacement vectors that incorporate the counterselectable sacb marker a lacz alphaallele for bluewhite screening the puc vectors multiple cloning site with unique restriction sites an orit for conjugationmediated plasmid transfer and carbenicillin gentamicin gm and tetracycline selectable markers a cassette was constructed that contains a gmr selectable marker next to the green FL protein structural gene with both markers being flanked by flp recombinase target frt sites the frt cassette was used to insertionally inactivate the cloned p aeruginosa pabc gene encoding aminodeoxychorismate lyase after conjugal transfer into p aeruginosa plasmid integrants were selected and deletion of unwanted dna sequences was promoted by SU counterselection the frt cassette was excised with HF close to from the chromosome after conjugal transfer of a flp recombinaseexpressing plasmid this sacbcontaining plasmid was subsequently cured by sucrose counterselection resulting in an unmarked p aeruginosa delta pabc strain

69|120|128

fluorescent|sucrose|high frequencies

tubulins are highly conserved multidomain proteins that have to interact with eukaryotic chaperonins to gain their correct threedimensional conformation the prokaryotic chaperonin system of groeles is able to generate intermediate folding states but not natively folded tubulin to create a system for studying these folding intermediates tubulins from the giant amoeba reticulomyxa filosa alpha and beta tubulin were expressed in escherichia coli singly or in tandem in all cases soluble tubulin was generated in amounts of mgl culture this is the first reported expression of soluble tubulin in bacterial cells of particular interest was the observation that upon coexpression with r filosa beta tubulin proteolytic degradation of alpha tubulin was reduced and more fulllength product remained IN this observation points to a specific interaction of alpha and beta tubulins in the e coli cell the sites of interaction are most probably the same that are responsible for the binding of native alpha and beta tubulin the established expression system therefore seems well suited for further studies concerning the folding of tubulins

116

intact

the cjun nterminal kinases jnks also called SAP kinases sapks belong to the mitogenactivated protein kinase mapk gene superfamily like all the mapks jnks are G1 through dual phosphorylation of a theronine residue and a IMT residue by a dual specificity kinase such as jnkkmkksek here we report the molecular cloning and characterization of hjnkk alpha a human homolog of the recently reported murine mkk alpha hjnkk alpha belongs to the mapk kinase gene family and is expressed in many adult tissues it is nearly identical to a recently reported human jnkk at the KD but with L1 differences in both amino and carboxylterminal sequences suggesting that hjnkk alpha may be an alternative spliced form of this kinase expression of hjnkk alpha but not its related kinases jnkkmkksek mek mkk or mkk leads to strong activation of jnk in several cell lines no activation of erk or p kinases was observed with this kinase an invitro kinase assay demonstrated that jnk activation by hjnkk alpha requires phosphorylation of the theronine and tyrosine residues at positions and in jnk furthermore hjnkk alpha activated the jnkdependent ST pathway in vivo by induction of cjun and atfmediated gene transcription in conclusion we have cloned the human homolog of murine mkk alpha which may be an alternative spliced form of human jnkk involved in transducing TPS upstream signals to regulate jnk activity in vivo

7|25|35|94|97|184|221

stressactivated protein|activated|tyrosine|kinase domain|major|signal transduction|specific

translocations involving the putative protooncogene mtgeto on q are frequently found in acute myeloid leukemia to date little is known of the genomic organization of this gene here we report that the mtg gene consists of exons distributed over kb of genomic dna two polymorphic microsatellite repeats are described including one in intron three alleles heterozygosity and another in the utr alleles heterozygosity expression of mtg was detected in a variety of normal human tissues with the highest mrna levels occurring in brain and HR previously two mrna forms produced by the alternative usage of the first exon have been reported we now describe a novel abundantly expressed alternatively spliced transcript resulting from the inclusion of a bp exon designated a that changes the reading frame and introduces a premature stop codon identical alternatively spliced mrna SCV were found to be produced by the highly conserved rat gene cbfat in the mouse suggesting an evolutionary significance

84|136|146

heart|variants|homologous

the general AS response of the grampositive soil bacterium bacillus subtilis is controlled by the sigma b transcription factor sigma b activity is regulated by the newly discovered partner switching mechanism of ST which integrates the two different classes of challenges which posttranslationally activate sigma b environmental stress and energy AS our investigation of a possible sigma b homologue in the related soil bacterium b licheniformis had two goals first this study would contribute to understanding the distribution of the sigma b general stress system among grampositive bacteria second a phylogenetic comparison of regulatory systems can supplement genetic and biochemical analysis by revealing conserved features that are critical for function we report here that b licheniformis cells contain a protein that closely resembles b subtilis sigma b in size and antigenic properties the level of this potential sigma b homologue rapidly increases following environmental or energy stress and the b licheniformis genome encodes a homologue of the sigb GA stress operon including the sigma b structural gene and seven rsb regulatory genes based on these results b licheniformis possesses a general stress system likely regulated by two coupled partner switching modules that sense and integrate the two broad classes of activating stress signals

2|32|50|158

stress|signal transduction|stress|general

in the nonmotile mycelial organism streptomyces coelicolor a the sporulation gene whig encodes a protein that closely resembles rna polymerase sigma factors such as sigma d of Bs which mainly control motility and chemotaxis genes here we show that the whig gene product purified from an escherichia coli strain carrying an expression construct could activate e coli core rna polymerase in vitro to transcribe a sigma ddependent motilityrelated promoter from b subtilis such rna polymerase holoenzyme preparations could also transcribe from an s coelicolor promoter pth previously shown to require an intact whig gene for invivo transcription the invivo dependence on whig was therefore shown to be direct unusually the initiation of pth transcription in vitro depended on the provision of appropriate dinucleotides the whigdependent pth transcription unit consisted of a single gene orfth sequence comparisons suggested that the gene product was a member of a small group of proteins that include the b subtilis and e coli prox proteins though none of these proteins shared more than about of D2 primary sequence identity they had similar size and hydropathy profiles and could be aligned end to end to reveal a mosaic of similarities the prox proteins of b subtilis and e coli are implicated in glycine betaine transport in response to hyperosmotic stress however disruption of orfth did not cause any obvious phenotypic changes in growth or development on media of varying osmotic strengths

27|170

bacillus subtilis|extended

sulfite reductase sir represents a key enzyme in sulfate assimilation in higher plants the genomic dna sequence of the sir gene from arabidopsis thaliana including regulatory and structural regions was isolated and characterized the CS of a kb fragment encoding sir revealed a coding region of basepairs bp that consists of eight exons separated by seven introns between and bp in length the transcription start point was determined bp upstream of the translation start site southern analysis indicates a single locus for the sir gene that gives rise to a kb mrna in leaves and in roots the promoter region was verified by functional expression of the gusa reporter gene in transgenic a thaliana plants and was shown to provide correct expression in root and leaf

34

sequence

to determine whether protein levels of interleukin il in induced sputum reflect the degree of eosinophilic inflammation we evaluated the role of il on clinical characteristics in stable AP il level differential eosinophil count and level of eosinophil cationic protein ecp in IS were all significantly higher for asthmatics than for normal controls both eosinophil counts and ecp levels in induced sputum were inversely correlated with the degree of airflow limitation fevfvc in addition patients with measurable il in sputum had significantly more eosinophils higher C2 of ecp in sputum and lower fev percent predicted than did patients with levels of il beneath the limit of detection however we found no significant difference in il levels between atopic and nonatopic asthmatics il level in induced sputum is a good indicator of eosinophilic inflammation in atopic and nonatopic asthmatic patients

28|42|85

asthmatic patients|induced sputum|levels

two methods of defining perception of asthmarelated changes in airflow were compared and relationships to clinical opinions of severity and assessments of psychological functioning were investigated perceived breathlessness vab and peak expiratory flow pef were recorded by subjects for days perception was defined by correlation of the two values and by the ratio of maximum NC in pef and related change in vab the latter method defined poor perceivers pp and exaggerated perceivers ep in whom presence of a psychological disorder was high of pp of ep compared to of the remainder clinical severity in ep was assessed as greater than appeared warranted

55

change

we examined the effect of ambroxol on superoxide anion production before and generation after phorbolmyristate acetate pma stimulation of lung alveolar macrophages lung free cells including lung alveolar macrophages were obtained from fischer rats and guinea pigs using bronchoalveolar lavage the superoxide anion produced by lung alveolar macrophages with or without stimulation of pma was measured by lucigenindependent CL method using a photon counter ambroxol inhibited the superoxide anion production and generation by lung alveolar macrophages harvested from both f rats and guinea pigs in a dosedependent fashion approximately mumoll of ambroxol inhibited of O2- of lung alveolar macrophages in rats and guinea pigs whereas a slightly greater dose of ambroxol ie mumoll was necessary to inhibit of pmaenhanced superoxide generation by lung alveolar macrophages these results suggest that ambroxol acts as an antioxidant in mu lungs and may be a potential therapeutic option for ROS speciesassociated lung disorders including bronchial asthma

58|94|135|145

chemiluminescence|superoxide production|murine|reactive oxygen

inhaled medication is commonly prescribed for the treatment of asthma and chronic obstructive pulmonary disease copd but is often not properly used by patients a total of patients suffering from asthma or copd took part in a study that evaluated how patients utilized their metereddose inhaler mdi or dry powder inhaler using a standardized inhaler checklist two hundred eightyone patients made at least one mistake in the inhalation technique the mistakes were classified into skill and nonskill mistakes two hundred patients made one or more skill mistakes and patients only made one or more nonskill mistakes the most common skill error was not continuing to inhale slowly after activation of the canister the nonskill item most patients had difficulties with was exhale before the inhalation patients who used an mdi made significantly fewer nonskill mistakes than patients using a dry powder device older patients had more difficulty with the correct use of the inhaler than younger patients there was no difference in PE between men and women in this patient sample most patients failed to use their inhaler correctly regular instructions and checkups of inhalation technique are the responsibility of the physician and should be a standard and routine procedure

162

errors

asthma is one of the most common chronic diseases the prevalence of asthma appears to be increasing national data have suggested that oregons asthma MR has been twice as high as the national average the prevalence and impact of asthma are not systematically assessed or tracked within oregon we estimated the current asthma prevalence and mortality using data from statewide hospital discharge data the behavioral risk factor surveillance survey and school registration data from the portland area a conservative estimate of oregons current asthma prevalence is and lifetime asthma prevalence is estimated at for both adults and children it appears that current asthma prevalence increases during adolescence the willamette valley an area known for high pollen counts does not have a higher rate of hospital discharges per capita than other areas within the state oregons apparent higher asthma mortality may be the result of the oregon state health divisions aggressive policies toward accurate reporting on death certificates oregons age distribution may also contribute to the apparent higher asthma MR

24|168

mortality rate|mortality rate

there is a paucity of information on the prevalence of asthma in children in india some evidence suggests that asthma is less common in developing than in the developed countries the present T0 was carried out to estimate its current magnitude in children in delhi the questionnairebased study was carried out in two randomly selected schools in delhi all the children were eligible the age range was years the questionnaires were distributed to all the children present n to be answered by either parent the key questions were related to complaints of recurrent wheezing in the past in the last year and also wheezing exclusively induced by exercise or colds in all questionnaires were completed and returned response rate there was a slight excess of males the prevalence of current asthma was and past asthma was reported by of children giving a cumulative prevalence of exclusive exerciseinduced asthma was and that associated with colds the current prevalence of all wheezing was thus and cumulative prevalence was while there was no sexrelated difference in prevalence wheezers were the highest in the year age group a significant association was found between the prevalence of wheezing and a family history of asthma odds ratio and presence of smokers in the family odds ratio when both the above factors combined the odds ratio for risk of asthma was there was no significant association with any economic class only of asthmatics had been labeled so by their physicians the prevalence of bronchial asthma and wheezing in children in delhi is quite high and comparable to that reported from several developed countries a positive family history of asthma and presence of smokers in the family emerged as significant risk AF

32|283

study|factors

the handling of the babyhaler spacer for inhalation therapy in children years old with asthma was tested and the perceptions of the investigators with the instruction of this device to the parents was investigated it was an open multicenter study of patients with a treatment period of weeks and a week followup period thirty pediatricians in dutch local and university hospitals participated using a standardized instruction the purpose and the application of the babyhaler were explained and demonstrated and parents were asked to use the babyhaler on their own child the quality of the demonstration was scored and if necessary additional instruction was given in the following treatment period of weeks record card was completed the convenience score of the babyhaler and the symptom score were recorded at the following evaluation visit the handling of the babyhaler by the parents was reevaluated eightynine percent of the parents could use the babyhaler correctly after one instruction the instructions of the babyhaler were considered to be easy in of the cases by the parents and the investigators during the treatment period the handling score impossible to use very easy improved from to at the evaluation visit the correct use was similar to the result at the instruction visit no differences were found in the ease of handling between the three age CG and years the application of inhalation therapy via the babyhaler was favored by of the parents as compared to the former treatment method we demonstrated that the babyhaler is easy to use by the parents and that the instruction of the device by practitioners is also perceived as easy

219

groups

the indications for transfusion have never been evaluated in an adequately sized CT a pilot study was conducted to plan larger clinical trials

12

clinical trial

in the event of hemorrhage and BL platelets play a vital role in the coagulation process however there are currently no acceptable protocols for longterm storage of platelets as a first step toward testing the efficacy of stored platelets or PLT substitutes in vivo a flow cytometric technique was developed to detect human platelets in rb blood

6|40|55

blood loss|platelet|rabbit

trials evaluating granulocyte transfusions for therapy of febrile neutropenia demonstrated mixed clinical results which may in part be due to the low PMNL cell dose of the transfused components to optimize the latter variable the effect of various interface offset io settings during apheresis on granulocyte collection efficiency gce and yield was determined in normal donors given granulocytecolonystimulating factor gcsf

22

neutrophil

during the use of commercial red cell rbc acidelution kits for adsorption and elution adsorptionelution studies with antid unexpected reactive eluates Rh were obtained from d rbcs such results were not obtained with a parallel xylene method or historically with heat and ether methods

21

antid

two cases of SH a among persons exposed to the same lot of solventdetergenttreated antihemophilic factor viii concentrate were reported to a surveillance system an investigation was conducted to find additional cases and determine the source of infection

3

hepatitis

on february in italy a law number referred to as was promulgated providing economic indemnity for persons INF with hepatitis b virus hepatitis c virus and hiv via transfusion or the administration of hemoderivatives

17

infected

cells from patients with fanconi anemia fa show decreased viability and decreased chromosome stability after treatment with dna crosslinking agents compared to normal cells fa cells also show a relative accumulation at the gm transition T3 such treatment this has suggested a possible checkpoint abnormality in the studies presented here treatment with hydroxyurea caffeine or inhibitors of cell cycle kinases did not reveal abnormalities in survival or chromosome stability in faa or fad cells chromosomal breaks introduced by hydrogen peroxide or methyl methanesulfonate accumulated to the same extent in faa or fad cells as in normal cells we conclude that faa and fad cells respond normally to agents known to alter the cell cycle or introduce dna strand breaks fa cells process strand breaks and a variety of dna monoadducts normally our results are compatible with repair of dna crosslinks being slower in fa than in normal cells and fa cells having normal cell cycle checkpoints

35

after

chromosomal fragile sites are regions that are intrinsically unstable and are susceptible to experimentally induced damage in most cases the target and mechanism of induction of fragility are unknown using ectopic integration of engineered dna arrays to create new fragile sites we and others have previously shown that the transcriptionally competent u gene is necessary and sufficient for induction of fragility at the rnu locus upon infection of human cells with adenovirus in the present study we have investigated the response of the rnu locus to cytosine arabinoside arac an inhibitor of dna polymerases and a common inducer of fragile sites we demonstrate that the rnu locus is sensitive to the drug and that aracinduced fragility is dependent upon a functional u gene and on the expression of the cellular p protein our results identify a novel dna structure associated with fragile CS and suggest a role for transcription and repair processes in rnu fragility

142

sites

chemotherapy is frequently limited by the development of multidrug resistance a major cause of which is activation of the pglycoproteinencoding mdr gene we have previously developed a pglycoproteinexpressing multidrug resistant subline hle from the nonpglycoproteinexpressing human hl promyelocytic leukemia cell line a possible cause of mdr silencing in hl cells is methylation of the promoter proximal region thus demethylation occurring as a result of drug treatment may be responsible for mdr activation in the multidrug resistant subline using the bisulphite genomic sequencing technique we demonstrated that hl dna is methylated at multiple CS within two distinct areas one UP and one downstream of the transcription start point only a single site in each area was methylated in all strands examined with the remaining adjacent sites showing partial methylation in contrast dna from the multidrug resistant hle subline was unmethylated at essentially all sites in both areas thus the development of the pglycoproteinexpressing multidrug resistant subline was associated with demethylation of the mdr region

92|98

sites|upstream

in mammalian cells nonhomologous endjoining is the predominant mechanism to eliminate dna double strand breaks such events are at the origin of deletion mutagenesis and chromosomal rearrangements the hallmark of fanconi anemia an inherited cancer prone disorder is increased chromosomal breakage associated to overproduction of deletions knowing that double SB are at the origin of deletion mutagenesis the question arises whether their processing is affected in fa we set up a host cell endjoining assay to analyze the fate of double strand breaks into extrachromosomal substrates transiently replicated in normal and fad lymphoblasts although no difference in plasmid survival was found bluntended breaks were sealed with significantly lower fidelity in fa cells resulting in a higher deletion frequency and a larger deletion size the results suggest that fad and fab gene products are likely to play a role in endjoining fidelity of specific dna double strand breaks

49

strand breaks

we have applied exon amplification grail exon prediction and est database searching to a mb segment of chromosome p exp and computational methods of identifying exons were comparable in efficiency and apparent false positive rate but were complementary in gene identification revealing distinct overlapping sets of expressed sequences est searching was most powerful when we considered only those ests that show evidence of splicing relative to the genomic sequence the combination of the three gene finding methods produced a transcription map of loci in this segment of p that includes known human genes homologs of loci identified in rodents and several anonymous transcripts including a putative novel dna polymerase and a gene related to drosophila ash while most of the genes in the region have been found our data suggest that even with the entire dna CS available complete saturation of the transcript map will require additional focused experimental effort

19|136

experimental|sequence

todays society is more litigant in accidental injury occurrences with many injuries involving children malpractice suits have resulted in an influx of information and rehabilitation supports to address the growing need for legal representation in life care planning for injured persons this easily understood tool provides the injured party with a viable RPA to community interdependence and inclusion contrary to the conventionally medical model where the person with a disability is viewed as needing fixed this approach examines culture community and the long term effects of disability the individual with a disability is seen and understood within four aspects of personhood capacities gifts and passions inclusive relationships personal involvement and understanding their SC and community this life care planning process examines five eras in ones life cycle formativeschool age years early work T2 work years and retirement years the specific needs of the individual in each life era are examined and specific costs itemized the provision of adequate services and supports affords tremendous opportunities and inclusion into the community

52|112|132

approach|culture|years later

whether children with cerebral palsy cp differ from nonhandicapped nh children was studied using bodyscaled information about the passability of a doorwaylike opening there were children of whom were nh and had cp the latter consisting of who could stand and walk unaided cpwalk and confined to a wheelchair cpwheel all groups were divided into two age ranges and year and were tested on two occasions separated by an interval of months on both occasions testing involved three consecutive tasks making perceptual prejudgments about the passability of an opening varying in width actually attempting to locomote through the openings performance and another round of postjudgments judgments and performance were expressed in terms of absolute and relative outcomes the latter differing from the former in accounting for differences in body width relative to aperture width there were two main findings firstly the younger cpwalk children were the least able to employ bodyscaled information in judging aperture width and the older cpwheel and nh the most able secondly when performances in passing through the openings were adjusted for differences in body width all groups had similar outcomes this findings lends credence to the notion that when action is used in the service of perception this is beneficial for the visualspatial abilities of both nh and cp children the T0 concludes by pointing out future directions in this type of research as well as some of the clinical implications of the findings

216

study

static standing balance is commonly measured with research laboratory systems labsys or clinical systems clinsys the purposes of this study were to assess the reliability of two systems designed to measure static standing balance in nondisabled children compare the findings derived from the two systems of measurement and examine the relationship between anthropometric measures and postural sway twentyfive ND children male female ages year months to years months mean years months sd years months participated in the T0 each child stood on the labsys and the clinsys for three consecutive second measurement periods intraclass correlation coefficients icc for the three trials on each system were labsys and clinsys the level of agreement between the two systems was icc younger children exhibited more R2 and less agreement between measurement trials using the clinsys however older children demonstrated more similar sway indices when comparing the two systems of measurement twoway analysis of variance indicated that there were significant differences between sway indices measured by the two systems p and between the youngest children aged years and all other children p in addition agreement among trials for the two systems was different depending on the age group measured rho coefficients for sway index and age height weight and foot length ranged from to for the labsys p and to for the clinsys p stepwise multiple regression analysis indicated that height was the most significant predictor of sway when measured by the clinsys r p whereas age was the most significant predictor when sway was measured using the labsys r p the results suggest that the degree of postural sway and the reliability of the measurement itself are influenced by the age of the child and the measurement system employed

58|77|122|195

nondisabled|study|variability|correlation

development of an instrument to measure modifications of motor quality in handicapped children receiving PT

14

physiotherapy

cognitive rehabilitation therapy is a systematic effort to assist brain damaged individuals in developing ways to compensate for cognitive deficits although this treatment is considered standard in a field of rehabilitation access to services is limited by the availability of specialists a MM for bringing cognitive rehabilitation techniques to children with acquired brain injury within their own school setting is presented a convenience sample of public high school students with acquired brain injury received biweekly cognitive rehabilitation sessions for weeks treatment was based on a developmental model of brain functioning treatments were provided by TR school teachers under the supervision of psychologists specializing in cognitive rehabilitation students were evaluated pre and posttreatment using neuropsychological tests after treatment the students demonstrated a significant increase in general memory ability p these gains were due mostly to increases in verbal learning ability integration of cognitive rehabilitation therapy with public school services is a significant step in providing this valuable treatment to a wider range of children with acquired brain injury

42|94

model|trained

a year old boy with epilepsy and learning difficulties is reported at years of age he was diagnosed with common ALL leukaemia and received therapy according to the uk protocol ukall viii this included prophylactic cns radiotherapy and chemotherapy he did not develop cns leukaemia and CR remission was achieved at age he began to experience lethargy and learning difficulties especially problems with handwriting concentration and SM furthermore he began experiencing atypical absence seizures which were provoked by concentration at times of tiredness eeg showed BL nonspecific abnormalities with some epileptiform features over the following years several antiepileptic drugs were prescribed although with the changes in therapy initial remissions have been achieved the seizures have each time continued to relapse at age eeg was very abnormal showing frequent generalized slow or sharp waves at age mri revealed multiple discrete small highintensity lesions in the subcortical VWM of both hemispheres problems with lethargy concentration and SM persist and although multiple antiepileptic drugs have been prescribed seizures continue to occur almost daily

20|46|66|85|145|154

acute lymphoblastic|complete|memory|bilateral|white matter|memory

the present case study provides evidence that a specific deficit can be improved by a treatment specific intervention in a boy who was prenatally exposed to dilantin and presented with a learning disability T3 implementation of a focused intervention for reading standardized test scores revealed that the child demonstrated a dramatic improvement in reading performance but remained consistent in other academic areas this indicated that his reading improvement was the direct effect of specific training in reading but did not generalize to other content areas this finding is significant because it provides support for treatment specificity in contrast to treatment generalizability

33

after

premenstrual dysphoric disorder pmdd has remained in appendices of the last two editions of the diagnostic and statistical manual of mental disorders due to lack of empirical study items included in its set of research criteria are considered tentative pending evidence of diagnostic validity the present study attempts to establish the construct validity of the pmdd criteria using the rasch method to analyze the validity of individual items as contributors to the diagnosis in contrast to the usual but less precise approach of using an external validator to establish the diagnostic utility of psychiatric conditions analysis of which items best differentiate participants with and without pmdd provides an idea of the relative ability of these items to distinguish pmdd it is recommended that the Az of angerirritability depressed mood and problems in interpersonal functioning be expanded in further studies and corresponding items added to symptom checklists

124

areas

this study examines the stability of a PCS disability construct across instruments and samples the purpose is not to report a formal equating of instrument calibrations but to indicate whether such an effort would be likely to succeed theory the economics transforming health care from its orientation toward crisisdriven disease reactions to population and evidencebased preventive health management and individualized disease management demand general scalefree measures of functional independence

7

physical

figure skating uses the median rank aggregation system for determining medal winners unfortunately the system can be influenced by idiosyncratic ratings made by some judges unlike the skating ability measures from a manyfacet rasch analysis these measures are constructed to be independent as statistically possible of item difficulties judge severities and the rating scale structure a manyfacet rasch analysis was conducted on data from the controversial ladies event at the olympics the results illustrate how the idiosyncratic ratings of the judges were not accounted for by the median rank system thus biasing the selection of the gold medal winner all sports that rely on judges ratings should investigate the use of a manyfacet rasch MM in order to bring more objectivity and fairness to the winner selection process

114

model

rasch analysis was used in this paper to evaluate the motor component of the fone fim the telephone version of the FIM fim for this purpose patients discharged from an inpatient geriatric assessment and rehabilitation program were assessed by TR research assistants using the fone fim the results at weeks postdischarge were compared to the observation fims obs fims done at home weeks postdischarge these patients had an average age of years and presented with multiple complex medical problems and significant functional decline the fone fim and the obs fim were shown to share a strikingly similar item hierarchy based on rasch item difficulty measures only bladder management and climbing stairs were misfitting items as indicated by item fit statistics the same item set and point scales were shown to be psychometrically optimal for both the fone fim and the obs fim based on the person separation index further research is required to address the issue of the optimal item set and scale C2 from psychometric and clinical perspectives

21|39|163

functional independence measure|trained|levels

the purpose of this T0 was to compare the results and interpretation of the data from a performance examination when four methods of analysis are used methods are traditional summary statistics interjudge correlations generalizability theory and the multifacet rasch model results indicated that similar sources of variance were identified using each method however the multifacet rasch model is the only method that linearized the scores and accounts for differences in the particular examination challenged by a candidate before ability estimates are calculated

4

study

the motor activities of daily living component of the fone fim the telephone version of the functional independence measure fim was evaluated in a cohort of patients who had been discharged to home from a geriatric inpatient assessment and rehabilitation program in the current T0 rasch person ability measures were derived from telephone assessments weeks after discharge and inhome assessments week later concordance between the modes was shown to be satisfactory for the rasch measures based on ICC coefficients however the telephone mode consistently generated lower estimates than did the observational mode this was due to the fact that the telephone mode underestimated motor function for the majority of patients who were at higher levels of cognition and motor function but overestimated for patients who were at lower levels of cognition and motor function at the item level concordance as determined by kappa statistics was better when the fone fim responses came from the patient rather than proxy respondents and when the assessments were done by more experienced rather than less experienced raters based on these findings a mixed strategy the telephone mode for patients capable of responding to the fone fim and inhome assessments for those who are incapable is recommended

44|77

study|intraclass correlation

the purpose of this T0 was to determine the optimal categorization of a SE orderedresponse scale using the rasch analysis and compare the performance of the rasch statistics and parameter estimates with conventional statistics a item scale to measure psychomotor SE was po to a total of children including boys and girls the data analysis started by collapsing the original five adjacent categories into two three and four categories and a total of data sets were derived each of these data sets including the original one was analyzed using the rasch rating scale model and a set of rasch modeldata fit category and separation statistics and parameter estimates as well as three conventional statistics were computed and compared it was found that instead of the fivecategory construct designed the best order of category meanings of the scale in respondents perceptions was a threecategory construct the rasch threshold estimates were sensitive indexes in determining the order of the categorization and that item separation statistics were useful in determining the optimal categorization T3 its order was confirmed the commonly used coefficient alpha was found not helpful at all in determining the optimal categorization the rasch analysis was demonstrated to be a useful posthoc analytic approach in determining the optimal categorization of an orderedresponse scale

4|13|40|42|170

study|selfefficacy|selfefficacy|administered|after

the sexual experiences survey koss gidycz wisniewski is a commonly used instrument for assessing various degrees of sexual aggression and victimization among male offenders and female victims rasch analysis was used to transform qualitative raw score observations into objective linear measures using the responses of a national sample of higher education men and women across the united states aged this paper supports the construct validity of the survey through DUE of the item hierarchy fit statistics and separation indices findings confirm a dimensional perspective on rape suggesting that sexually aggressive behaviors can be scaled along a single continuum from normal to extreme sexual behavior the item hierarchy reveals an arrangement of sexually aggressive acts in an order of mild to severe which compares with the one theorized by the authors of the ses identity plots demonstrate the validity of using a common set of ses item calibrations to measure both male and female respondents for interpretation of person responses to the ses three conclusions are suggested first rasch analysis must be employed to examine item responses effectively second when the survey is po to a college sample aged the item calibrations obtained in this paper can be used to measure offenders and victims third a total raw scoretomeasure conversion is not always sufficient to interpret person measures instead a scalogram method needs be added to the rasch analysis to separate the measures of offenders and victims who CR the survey implications for future research are discussed

69|182|237

evaluation|administered|complete

this study equates the physical SF-36 subscales of the medical outcomes study short form sf and the louisiana MSU health status instruments lsu hsi data from the sfs item physical functioning scale the pf and the lsu hsis item physical functioning scale pfs were fit to separate and mixed rasch rating scale models data were provided by a convenience sample of patients waiting for appointments in a public hospital general medicine clinic difficulty estimates for a subset of similar items from the two instruments were highly correlated indicating that the items from the two scales are working together to measure the same variable the measures from the two equated instruments correlate when disattenuated for error of the two instruments the pfss error is lower model fit is better and reliability coefficients are higher both instruments measure PCS SF-36 and can do so in a common unit of measurement conversion tables are provided for transforming raw scores from either instrument into the common metric

5|18|136|137

functioning|state university|physical|functioning

the psychometric properties of the oars adl scale comprised of seven physical MICs of daily living padl and seven instrumental activities of daily living iadl items were examined using a rasch measurement approach two of the padl items failed to demonstrate acceptable goodnessoffit with the measurement model but the remaining items could be combined into a single measure of adl ability although the oars adl scale was designed to identify those communitydwelling elderly who need supports and services to continue to live in the community the scale items were found to be poorly targeted to communitydwelling elderly since almost half of our sample received maximal scores rasch analysis identified how we might improve the sensitivity of the oars adl scale but its utility in outcome and longitudinal studies remains questionable

12

activities

this paper presents an analysis of differential item functioning dif in a certification examination for a medical specialty the groups analyzed were physicians from different subspecialties within this area and physicians who qualified for the examination through two different experiential pathways the dif analyses were performed using a simple rasch MM procedure the results were shown to be readily interpretable in terms of the known differences between the CG being compared these results serve as validity evidence for the rasch model procedure as a means for evaluating dif in examinations the conclusion is drawn that complex procedures are not required to generate interpretable results if relevant differences between the groups being compared are known this suggests that the inability of many researchers to interpret results for racialethnic or gender groups is not due to inadequacies of the methods but more likely to lack of pertinent knowledge about group differences

50|68

model|groups

in order to apply the rasch MM to multiplechoice items incorrect responses to distractors are usually aggregated to a single category in doing so information of individual distractors disappears in this paper a raschtype analysis is proposed where one parameter is assigned to each distractor the information is thus preserved the proposed distractor model can be applied to investigate the performance of distractors which is useful for item revision this model is a necessary condition of the rasch model that is fitting the distractor MM will fit the rasch model but not vice versa the results of a small simulation T0 show that parameter REC of the distractor model is very satisfactory a real data set of twenty multiplechoice items was analyzed some items were found to fit the rasch model rather than the distractor model it is this diagnostic value that makes the distractor MM suitable for multiplechoice items

6|84|100|104|145

model|model|study|recovery|model

the functional assessment measure fam has been proposed as a measure of disability in postacute traumatic brain injury tbi outpatients it is comprised of the items of the FIM fimsm scored in terms of dependence and of newly designed items scored in terms of dependence items or performance items the fimsm covers the domains of selfcare sphincter management mobility locomotion communication and social cognition the new items explore the domains of community integration emotional status orientation attention readingwriting skills swallowing and speech intelligibility by addressing a set of problems quite specific for tbi outpatients the fam was intended to raise the ceiling of the fimsm and to allow a more precise estimate of their disability these claims however were never supported in previous studies we administered the fam to tbi outpatient months median from trauma rasch analysis rating scale model was adopted to test the psychometric properties of the scale the fam was reliable rasch item and person reliability and respectively two of the famspecific items were severely misfitting with the general construct and were deleted within the item refined fam scale new items and fimsm items still retained signs of misfit the fam was on average too easy the most difficult item a new one employability did not attain the average ability of the subjects also it was only slightly more difficult than than the most difficult fimsm item SM the fam does not seem to improve the fimsm as a far as tbi outpatients are to be assessed

28|230

functional independence measure|memory

as organizations begin to implement work teams their assessment will ultimately reflect compensation strategies that move away from individual assessment this will involve not only using multiple raters but also the use of multiple criteria team assessment using multiple raters and multiple criteria is therefore necessitated however this can produce differences in ratings due to the leniency or severity of the individual team raters this T0 analyzed the ratings of individual members on different teams across different criteria of team performance utilizing the manyfacet rasch model statistical differences between the teams and criteria were calculated

65

study

to test transdifferentiation of retinal pigment epithelial rpe cells in suspension culture chick embryonic rpe sheets and dissociated rpe cells were cultured for two months in a nonadherent dish for suspension culture rpe cells isolated as a sheet aggregated immediately and remained the same size with their differentiated characteristics for two months the presence of basic fibroblast growth factor bfgf at concentrations of ngml or higher induced the formation of a spherical lentoid structure which was positive for crystallin and bfgf receptor in contrast dissociated rpe cells did reaggregate but did not develop the lentoid structure even in the presence of bfgf the transdifferentiation of rpe cells to the lentoid structure in this study was in contrast to their transdifferentiation to the retina as reported in a previous T0

128

study

to study the effect of PLV plv with perfluorocarbon on ARDS groups of rabbits were examined to compare after acute respiratory failure was induced by lung lavage with sea water in of the rabbits of the rabbits were treated with conventional mechanical ventilation ac group and of the rabbits were treated with plv using perfluorocarbon ap group the remaining normal rabbits without ARDS were treated with plv with perfluorocarbon as a control group pl group in the pl group pao paco blood ph pulmonary compliance or pathological findings were not so changed T3 plv in the ac and ap CG paco significantly increased and in contrast pao and pulmonary compliance significantly decreased T3 lung lavage however these findings improved to almost the same levels as those of a control group within h T3 the plv treatment in the ap group but in the ac group these gradually deteriorated over time as for the pathological findings pulmonary vascular congestion alveolar hemorrhage and inflammatory infiltration were observed in the ac group however these findings were not observed in the specimens of the ap group from these results plv with perfluorocarbon was shown to be useful to improve gas exchange and pulmonary functions without L1 SE

5|10|62|92|99|112|132|201|202

partial liquid ventilation|acute respiratory failure|acute respiratory failure|after|groups|after|after|major|side effects

steadystate serum concentrations of carbamazepine cbz and valproic acid vpa were investigated in normal weight body mass index bmi to lean smaller than bmi and moderately obese subjects greater than bmi who received either mgday of cbz or mgday of vpa the cbz serum concentration in lean subjects was significantly higher than that in normal weight subjects however no significant differences in vpa serum concentration were found between the three CG the cbz SS concentration decreased with increases in total body weight and the vpa serum concentration decreased with increases in ideal body weight however both SS concentrations were not correlated with bmi these results suggest that vpa doses should be calculated using ideal body weight and that degree of obesity may affect cbz SS concentration rather than vpa serum concentration

70|73|96|124

groups|serum|serum|serum

the clinical implications of a wider distribution of salivary type stype isoamylase activity as compared with that of pancreatic type ptype isoamylase activity in the serum of young female adults of years old was studied a high correlation existed between the stype isoamylase levels in the initial determination and those in the second determination one year after on the same subjects indicating that the wider distribution of stype isoamylase level reflects an individual R2 the SS level of stype isoamylase was highly correlated with the stype isoamylase activity in saliva among the additional AF studied a weak positive rho was present between energy NI and the total and stype isoamylase activities in serum however there was no negative correlation between the stype isoamylase level and body mass index bmi which was reported for young male adults

73|75|93|98|103

variation|serum|factors|correlation|intake

with advances in lectin affinity electrophoresis of alphafetoprotein afp the detection of significant changes in serum afp at low levels in cirrhotics has become important for early detection of hepatocellular carcinoma SS afp C2 of healthy individuals without abnormal CL function tests or virus markers of hepatitis b and c were determined by enzyme immunoassay with imxafp dainapack using automated imx apparatus set at twice the ordinary sensitivity and compared with those of individuals with abnormal LF tests andor positive hepatitis virus markers the coefficient of variation in this assay was less than at afp C2 as low as ngml with a lower PCD limit of ngml the afp level of healthy population showed a gaussian distribution curve after logarithmic transformation with a median and percentile reference range of ngml there was no significant difference in the afp level between males and females individuals with abnormal liver function tests CT showed no significant increase in serum afp unless they were associated with positive SH virus markers

31|33|39|76|95|103|149|163

serum|levels|liver|liver function|levels|detection|alone|hepatitis

we investigated the degree of local heat and swelling of the thigh muscles produced by exercise eleven university athletes aged from to years old performed isokinetic exercise of the right knee on a cybex ii then serial determination of thigh circumference and thigh temperature up to min after exercise as well as serial magnetic resonance mr imaging up to min after exercise was performed on both thighs the circumference of the right thigh peaked at min after exercise and returned to normal at min the temperature of the right thigh peaked at min after exercise and was not normalized after min except in two subjects tweighted mr images showed a marked increase in the signal intensity of the right knee flexor and extensor muscles the signal intensity peaked immediately after exercise and subsequently decreased gradually but did not return to normal after min in some muscles changes in the thigh circumference were closely correlated with changes in the mr findings the changes in the thigh muscles T3 knee exercise could be demonstrated using mr imaging thigh circumference and thigh temperature data these parameters may provide indicators for managing muscle fatigue and recovery

166

after

an exposure to gb virus chepatitis g virus gbvchgv was studied among populations at risk for blood and sexual SE to analyze risk factor of the transmission of the virus blood samples were drawn from intravenous drug users ivdu female highclass commercial SWs csw and male outpatients mop at a sexually transmitted diseases std clinic in chiang mai thailand these blood samples were analyzed for gbvchgv rna antibodies against second env protein of gbvchgv antie antihepatitis c virus antibody hcvab HBcAb hbcab and antibodies against human immunodeficiency virus hivab prevalences of gbvchgv rna antie hcvab hbcab and hivab were and in ivdu and in csw and in mop while the prevalence of gbvchgv rna was higher in ivdu than in csw and mop comparable prevalences of antie among the three populations were found intravenous drug injection showed association with gbvchgv rna while PH of std associated with antie in conclusion intravenous drug i.t. and std were found to be risk factors for the previous SE to gbvchgv but std did not increase the risk of the gbvchgv viraemia

19|42|70|80|142|152|164

exposure|sex workers|envelope|hepatitis b core antibody|history|injection|exposure

the crosssensitization to stereotyped SMB between mazindol mzd and methamphetamine map was investigated in rats mzd and mgkgday po map and mgkgday po and saline mlkg po were administered once daily for a week challenge with mzd mgkg po on the th day caused markedly stereotyped behavior in mappretreated group compared with the salinepretreated control group map mgkg poinduced stereotyped SMB on the th day was also greater in mzdpretreated group rather than the salinepretreated control group these results suggest that repeated mzd and map administration crosssensitizes to their stereotypeproducing effects

4|60

behavior|behavior

the aim of this T0 was to find suitable methods for basic anatomic evaluation of the supratubal recess and the anterior surface of the tensor fold

4

study

to avoid the limitations of CWD surgery yet maintain the SE provided by canal wall down mastoidectomy the authors have developed a completely reversible canal wall down mastoidectomy technique the purpose of this case report is to determine the feasibility of the reversible canal wall down mastoidectomy technique in the treatment of a patient with an aural cholesteatoma

5|10

canal wall down|exposure

this T0 aimed to evaluate improved surgical outcomes from middle and external ear surgery using local vascular flaps to enhance tissue healing

1

study