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PubMed2002

Somatic and embryonic cell nucleus transfer into intact and enucleated immature mouse oocytes.

BACKGROUND: The aim of our study was to evaluate the possibility of embryonic or somatic cell haploidization after fusion with intact or enucleated immature oocytes which were subsequently cultured in vitro. Embryonic or somatic cell nuclei do not undergo premature chromosome condensation when fused to intact or enucleated immature oocytes whose maturation is prevented by dibutyryl cyclic AMP (dbcAMP). The presence of dbcAMP permits, however, the completion of DNA replication in somatic cell nuclei. METHODS AND RESULTS: The chromosomes condensed when the reconstructed cells were released from the dbcAMP block. When somatic or embryonic nuclei were introduced into intact immature meiotically competent oocytes and subsequently cultured their chromosomes assembled on a common spindle with meiotic chromosomes and proceeded through the meiotic-like division, judged according to the presence of the first polar body extruded. When embryonic cell nuclei were introduced into cytoplasts obtained from immature meiotically competent oocytes, polar bodies were extruded in about 75% of reconstructed cells but the metaphase plates were abnormal in almost all cases. When somatic cell nuclei were inserted into the above cytoplasts, polar bodies were extruded only very exceptionally and in these cells chromosomes were arranged in abortive metaphase plates. CONCLUSIONS: Our results suggest that somatic cell nuclei are unable to proceed through the reduction division (haploidization) when introduced into an immature oocyte meiotic cytoplasm.

Animals↗
PubMed2005

Number of germ cells and somatic cells in human fetal ovaries during the first weeks after sex differentiation.

BACKGROUND: This study presents the number of germ cells and somatic cells in human fetal ovaries during week 6 to week 9 post conception, i.e. the first weeks following sex differentiation of the gonads. METHODS: One ovary with attached mesonephros from each of 11 individual legal abortions was used for estimation of cell numbers. After recovery of the fetus, the ovary-mesonephric complexes were immediately isolated, fixed and processed for histology. A stereological method was utilized to estimate the total number of oogonia in all ovaries and somatic cells in seven of them. RESULTS: The number of oogonia per ovary increased from approximately 26,000 in week 6 to approximately 250,000 in week 9 and somatic cells from approximately 240,000 to approximately 1.4 x 10(6). The ratio of oogonia to somatic cells tended to increase throughout the period. The concentration of oogonia was similar in the cranial (mesonephric connected) part and the caudal part of the ovaries. CONCLUSIONS: This is the first stereological estimation of the number of oogonia and somatic cells in human fetal ovaries, and the first estimation of germ cells and somatic cells in ovaries aged <9 weeks. The number of oogonia in week 9 is comparable to the numbers previously published based on non-stereological estimations. We found early stages of meiosis in fetal ovaries from week 9.

Cell Count↗
PubMed2007

Brief report: testing the factorial invariance of the CBCL Somatic Complaints scale as a measure of internalizing symptoms for children with and without chronic illness.

OBJECTIVE: To examine the factorial invariance of the Somatic Complaints subscale of the Child Behavior Checklist as a measure of Internalizing Behavior Problems across a sample of children with and without spina bifida. METHODS: Multisample confirmatory factor analysis was used to compare mother and father report on the Somatic Complaints subscale across a sample of children with spina bifida and a matched comparison sample of able-bodied children ages 8 through 11 years (N = 68 for mother report in each group; N = 54 for father report in the spina bifida group and 53 for the able-bodied group). RESULTS: Although there were no significant between-group differences in the magnitude of factor loadings, significantly more variance in scores on the Somatic Complaints scale was unrelated to Internalizing Behavior Problems for the spina bifida group, compared to the able-bodied group. There were no between-group differences when father data were analyzed, but the latent variable of Internalizing Behavior Problems explained little variance in the Somatic Complaints scale for either group. CONCLUSIONS: Maternal report of Somatic Complaints on the CBCL does not appear to measure Internalizing Behavior Problems in the same manner across groups of children with and without spina bifida. This suggests that the Somatic Complaints subscale should be interpreted with caution when measuring Internalizing Behavior Problems within this population.

Anxiety↗
PubMed2005

Disabling musculoskeletal pain and its relation to somatization: a community-based postal survey.

BACKGROUND: Somatization (a tendency to report distress from somatic symptoms) is a little studied, but potentially important, confounder and effect modifier in occupational studies of musculoskeletal disease. AIMS: To assess the role of somatization as a risk factor for disabling regional pain. METHODS: A questionnaire was mailed to 4998 subjects of working age. Questions were asked about chronic and disabling pain in the past 12 months affecting the arm, low back, knee or combinations of these sites. Distress from physical symptoms was assessed using elements of the Brief Symptom Inventory and mental well-being was assessed using the short-form 36 (SF-36). Associations were examined by modified Cox regression and expressed as hazard ratios (HRs) with 95% confidence intervals (CI). RESULTS: Among 2632 responders, 24% reported chronic pain and 25% disabling pain at one or several sites. Risk of chronic or disabling pain increased strongly according to the number of somatic symptoms reported as bothersome. For example, the HR for chronic upper limb pain in those distressed by > or =2 somatic symptoms in the past 7 days versus none was 3.9 (95% CI 2.9-5.3), and that of disabling upper limb pain was 5.8 (95% CI 4.1-8.3). Similar patterns were found for the low back and knee, and there was a gradient of increasing risk according to the number of sites with disabling pain. In comparison, associations with SF-36 mental well-being score were weaker. CONCLUSION: Somatizing tendency should be evaluated as a possible confounder or effect modifier in studies of occupational risk factors for musculoskeletal pain.

Adult↗
PubMed1986

Differentiation of oocyte- and somatic-type 5S rRNAs in animals.

In some amphibians and bony fishes, oocyte- and somatic-type 5S rRNA genes are expressed differently in oocytes and somatic cells. In order to determine at what stage of animal evolution this differential expression system appeared and how it is regulated, the sequences of oocyte and somatic 5S rRNAs from three invertebrates (sea urchin, sea hare, and silkworm) and two vertebrates (lamprey and chick) were analyzed. It was found that the oocyte 5S rRNA from lamprey consists of two components, while its somatic 5S rRNA consists of only one. In other animals, such differential expression of 5S rRNA in oocytes and somatic cells was not seen. A phylogenetic tree of 63 animal 5S rRNAs was constructed by means of the parsimony method, and the evolution of oocyte and somatic-type 5S rRNAs was discussed.

Animals↗
PubMed2006

Determination of genetic stability in long-term somatic embryogenic cultures and derived plantlets of cork oak using microsatellite markers.

Microsatellites were used to test genetic stability in somatic embryos (SE) of Quercus suber L. The SE were obtained by a simple somatic embryogenesis protocol: leaf explants from two adult plants (QsG0, QsG5) and from two juvenile plants (QsGM1, QsGM2) were inoculated on Murashige and Skoog (MS) medium with 2,4-dichlorophenoxyacetic acid and zeatin. Calluses with primary embryogenic structures were transferred to MSWH (MS medium without growth regulators) and SE proliferated by secondary somatic embryogenesis. High morphological heterogeneity was found among cotyledonary SE. However, converted plants looked morphologically normal with well-developed rooting systems and shoots. The genetic stability of the plant material during the somatic embryogenesis process was evaluated by using six to eight nuclear microsatellites transferred from Q. myrsinifolia Blume, Q. petraea (Matts.) Liebl. and Q. robur L. Five of eight microsatellites distinguished among the genotypes analyzed, and for QsG0, QsGM1 and QsGM2, uniform microsatellite patterns were generally observed within and between SE and the respective donor genotypes. For genotype QsG5, the same pattern was observed in all samples analyzed except one, where the mutation percentage was 2.5%. We conclude that microsatellite markers can be used to assess genetic stability of clonal materials and to determine genetic stability throughout the process of somatic embryogenesis. The simple somatic embryogenesis protocol described has potential for the commercial propagation of Q. suber because it results in a low percentage of mutations.

Alleles↗
PubMed2005

Microinjection of cytoplasm or mitochondria derived from somatic cells affects parthenogenetic development of murine oocytes.

Cloned mammals are readily obtained by nuclear transfer using cultured somatic cells; however, the rate of generating live offspring from the reconstructed embryos remains low. In nuclear transfer procedures, varying quantities of donor cell mitochondria are transferred with nuclei into recipient oocytes, and mitochondrial heteroplasmy has been observed. A mouse model was used to examine whether transferred mitochondria affect the development of the reconstructed oocytes. Cytoplasm or purified mitochondria from somatic cells derived from the external ear, skeletal muscle, and testis of Mus spretus mice or cumulus cells of Mus musculus domesticus mice were transferred into M. m. domesticus (B6SJLF1 and B6D2F1) oocytes to observe parthenogenetic development through the morula stage. All B6D2F1 oocytes injected with somatic cytoplasm or mitochondria showed delayed development when compared to oocytes injected with buffer. The developmental rates were not different among injected cell sources, with the exception of testis-derived donor cells injected into B6SJLF1 oocytes (P < 0.01). The developmental rate of B6D2F1 oocytes injected with buffer alone (98.8% survival) was different from those injected with somatic cytoplasm (60.8% survival) or somatic mitochondria (56.5% survival) (P < 0.01). Conversely, injection of ooplasm into B6D2F1 oocytes did not affect parthenogenetic development (100% survival). Our results indicate that injection of somatic cytoplasm or mitochondria affected parthenogenetic development of murine oocytes. These results have further implications for in vitro fertilization protocols employing ooplasmic transfer where primary oocyte failure is not confirmed.

Animals↗
PubMed2002

Neuropathy reduces viscero-somatic inhibition via segmental mechanisms in rats.

The effect of an experimental neuropathy on the viscero-somatic inhibition was studied in lightly anesthetized rats. In controls, colo-rectal distension at noxious intensities produced a multisegmental prolongation of the withdrawal response induced by noxious stimulation of the skin. In rats with a spinal nerve-ligation induced neuropathy this viscero-somatic inhibition was significantly reduced within the neuropathic segment (the hindlimb) but not outside of it (the tail). Naloxone, an opioid antagonist, attenuated this viscero-somatic inhibition in controls and it did not restore the inhibition in neuropathic rats. The results indicate that somatic neuropathy produces a segmental attenuation of viscero-somatic inhibition and this attenuation cannot be explained by a nerve injury-induced release of endogenous opioids. The decreased inhibition of somatic signals may contribute to the hypersensitivity observed in neuropathic conditions.

Animals↗
PubMedDate not supplied

Somatization in children and adolescents.

OBJECTIVE: To review the empirical literature on somatization in the pediatric age group, emphasizing prevalence, influence on health care utilization, issues in the development of somatization, comorbidity with other psychiatric disorders, assessment, and treatment. METHOD: One hundred nineteen studies and reports addressing medically unexplained somatic symptoms in children and adolescents were compiled via MEDLINE search and extensive cross-referencing. All available controlled studies were included, as were selected case reports and collections of cases. Selected citations from the adult literature were chosen for relevance to pediatric somatization. RESULTS: Recurrent, medically unexplained physical symptoms are common in the pediatric age group, are often associated with other psychiatric symptoms, and may represent a common presentation of psychiatric disorder in the primary care setting. DSM-III-R-defined somatization disorder is rare, and pseudoneurological symptoms are unusual. Patients may be at risk for potentially dangerous, costly, and unnecessary medical investigations and treatments, and they may excessively utilize health care services. CONCLUSION: Our current understanding of pediatric somatization and its consequences is limited. Collaboration between mental health professionals and primary health care providers is essential. Consistent terminology, developmentally appropriate classification, and systematic future research will be necessary for the development of successful prevention and treatment strategies.

Adolescent↗
PubMed1995

Antisocial behavior, somatization, and sensation-seeking behavior in mothers of clinic-referred children.

OBJECTIVE: The relationship between antisocial behavior and somatization was studied in a sample of mothers of clinic-referred, school-age children. The goal was to overcome some of the methodological limitations of past research in this area and to provide a preliminary test of the theory that these disorders share a common substrate: a tendency toward behavioral disinhibition. METHOD: Structured diagnostic interviews and an objective personality measure were used to assess antisocial behavior and somatization in a sample of 90 biological mothers (mean age 34 years) of children referred to an outpatient mental health clinic. A rating scale measure of sensation-seeking behavior was used as a measure of behavioral disinhibition. Structured interviews were used to assess a history of antisocial behavior in the children and their biological fathers. RESULTS: Results indicated a link between somatization and antisocial behavior both within individuals and across generations. This intergenerational link could not be solely accounted for by assortative mating between women with somatization and antisocial men. Both somatization and antisocial behavior were correlated with scores on the sensation seeking scale. CONCLUSIONS: These findings are consistent with the theory that antisocial behavior and somatization are related syndromes and that behavioral disinhibition may be a common predisposition that underlies both.

Adolescent↗
PubMed1997

Somatic symptoms in anxious-depressed school refusers.

OBJECTIVE: To identify the most common physical complaints in a sample of adolescent school refusers with comorbid anxiety and depressive disorders. Whether somatic symptoms are more likely to be associated with high levels of anxiety or high levels of depression was also explored. METHOD: Forty-four adolescents in a treatment study were evaluated at baseline with structured psychiatric interviews and measures of anxiety, depression, and somatization. RESULTS: The most common somatic complaints were in the autonomic and gastrointestinal categories. In simple regression analyses, anxiety level as measured with the Revised Children's Manifest Anxiety Scale and depression level as measured with the Beck Depression Inventory each significantly predicted the severity of somatic symptoms. The correlation between percentage of days absent from school and severity of somatic symptoms approached significance (r = .27, p = .074). CONCLUSIONS: Knowledge that somatic complaints are commonly an expression of underlying anxiety and depression may facilitate more rapid referral for psychiatric assessment and treatment and thereby help avoid unnecessary medical workups and sequelae from school refusal.

Adolescent↗
PubMedDate not supplied

Somatization disorder in sphincter of Oddi dysfunction.

OBJECTIVE: Sphincter of Oddi dysfunction (SOD) is a biliary disorder with a recognized pathophysiology and demonstrated surgical treatment. We sought to examine whether women with SOD were overrepresented on measures of somatization, sexual and physical abuse, socially compliant attitudes, and familial psychiatric illness. METHOD: We matched 33 women with SOD to 33 normal controls by age, sex, and race, and, with a case-controlled cross-sectional questionnaire, compared the groups on the study variables. Statistics included a chi 2 for categorical variables, t tests for scores of somatization and attitudes of social desirability, and Pearson correlation coefficients for post hoc associations of variables. Bonferroni corrections were used with chi 2 values to reduce capitalization by chance. RESULTS: SOD patients exhibited excessive nongastroenterological somatic complaints compared with controls (p < .0001). There was a statistical increase in reports of childhood sexual, but not physical, abuse in the SOD women (p < .02) compared with controls. The severity of the abuse correlated strongly with the severity of somatic complaints. There were no differences in social desirability attitudes or family psychiatric histories of the two groups. CONCLUSIONS: We conclude that SOD is associated with a high degree of somatization in adulthood, and a mean rate more than four times that of controls in self-reports of sexual abuse in childhood. The severity of childhood sexual abuse is correlated with the severity of somatization in later life. A psychological model for this disorder is suggested by the data. Increased psychiatric attention is indicated in the treatment of women with this disorder.

Case-Control Studies↗
PubMedDate not supplied

Elevated levels of psychophysiological arousal and cortisol in patients with somatization syndrome.

OBJECTIVE: This study investigates psychological and psychobiological processes in patients with somatization syndrome. METHOD: We compared physiological measures (heart rate, finger pulse volume, electrodermal activity, electromyography), cortisol levels, and subjective well-being during rest and during a mental stress task as well as selective attention and memory for illness-related words in 58 patients with somatization syndrome and 21 healthy controls. RESULTS: The somatization group had higher morning salivary cortisol concentrations, higher heart rates, and lower levels of finger pulse volume. During the mental stress task, patients with somatization syndrome felt more distressed and had higher heart rates, whereas controls showed habituation to the experimental situation. We were unable to demonstrate an attention or memory bias specific for somatization. CONCLUSIONS: The results point to several psychological, psychophysiological, and psychobiological mechanisms that might be involved in the maintenance of somatization syndrome. These results are discussed from a cognitive-psychobiological perspective.

Adolescent↗
PubMedDate not supplied

Association between somatic symptoms and 24-hour ambulatory blood pressure levels.

OBJECTIVE: To clarify whether somatic complaints in healthy normotensive men are associated with differential 24-hour blood pressure and heart rate measures. METHOD: Twenty-four-hour ambulatory systolic and diastolic blood pressure was monitored in 114 healthy normotensive men, aged 28 to 63 years, engaged in similar physical work. Means were calculated for each hour, for the whole 24-hour period, and for daytime, nighttime, and work time. Subjects were interviewed about somatic complaints, demographic data, and health habits, and body mass index was measured. RESULTS: After controlling for possible confounders, a positive association was found between the somatic complaint score and 24-hour, diurnal, and work-time systolic blood pressure (p = .014, p = .007, and p = .008, respectively). The association with casual systolic blood pressure was of borderline significance (p = .089). There was a positive trend, which did not reach statistical significance, in the relationship between somatic complaint score and all measures of diastolic blood pressure. Diurnal, 24-hour, and work-time heart rates were highest in the subjects with the highest somatic scores (p < .01 for all trends). CONCLUSIONS: In healthy normotensive men, somatic complaints are associated with an increased cardiovascular load. The effects of this increase on long-term cardiovascular morbidity and mortality are uncertain and warrant additional study.

Adult↗
PubMed1986

Excess medical care of women with somatization disorder.

We compared 50 women with somatization disorder to 25 women with major depression. The somatizers had three times as many operations and hospitalizations as the depressed women. Hospitalizations for headache and backache, abdominal and pelvic operations, and radiologic studies for abdominal complaints occurred frequently in the somatizers. Important factors in the excess medical care of these women were the difficulty in diagnosis and the unwillingness of physicians to use the diagnosis of somatization disorder to limit medical care. Even though somatization disorder has been well described in the medical literature for 30 years, somatizers still receive excess medical care.

Adult↗
PubMed1989

Oocyte and somatic tyrosine tRNA genes in Xenopus laevis.

Over a period of many months, Xenopus oocytes stockpile large quantities of tRNA for use during the first few hours of embryogenesis. To test the idea that these tRNAs are transcribed from one set of genes and that another set is used by somatic cells, we used synthetic oligonucleotides to analyze the sequence and steady-state levels of unspliced tyrosine tRNA precursors in Xenopus laevis oocytes, embryos, and cultured kidney cells. These analyses identify four kinds of tyrosine tRNA genes, two oocyte-type and two somatic-type, whose unspliced transcripts are distinguishable from one another by their different 5' leader and intervening sequences. The oocyte-type tyrosine tRNA precursors are present in oocytes, very abundant in gastrula embryos, but absent from postembryonic somatic cells. The somatic-type precursors are undetectable in oocytes but are found in gastrula and later stage embryos and in somatic cells. The major switch from oocyte-type to somatic-type transcripts occurs early during embryogenesis, between the midblastula transition and the onset of neurulation, but some oocyte-type precursors are also detectable in tadpoles.

Animals↗
PubMed1992

Morphoregulatory role of thidiazuron : substitution of auxin and cytokinin requirement for the induction of somatic embryogenesis in geranium hypocotyl cultures.

Somatic embryogenesis was induced in hypocotyl explants of geranium (Pelargonium x hortorum) cultured on media supplemented with various concentrations of N-phenyl-N'-1,2,3-thiadiazol-5-ylurea (thidiazuron). In less than 2 weeks, somatic embryos were observed in treatments containing levels of thidiazuron (TDZ) ranging from 0.2 to 1.0 micromolar. The use of N(6)-benzylaminopurine in combination with indole-3-acetic acid also evoked embryogenesis, but the efficiency of somatic embryo production was significantly lower than that obtained with TDZ. Hypocotyl culture for only 2 days on TDZ-supplemented medium before transfer to a basal medium was sufficient for inducing somatic embryogenesis. This distinction between the induction and expression of embryogenesis may provide an experimental system for studying the developmental biology of somatic embryogenesis. Substitution of the auxin-cytokinin requirement for the induction of somatic embryogenesis by TDZ suggests the possibility of a novel mode of its action by modulation of endogenous growth regulators.

Journal Article↗
PubMed1994

Evidence that the clonogenic cell in multiple myeloma originates from a pre-switched but somatically mutated B cell.

There is still much controversy about the precursor cell type in multiple myeloma (MM). Some authors claim that it is a pre-B cell, others state that it is a memory B cell or plasmablast. We have recently shown that the VDJ region of the MM immunoglobulin heavy chain gene is somatically hypermutated and antigen selected, without intraclonal variation or evolution in time. By using a patient-specific PCR approach we have now obtained evidence that the premyeloma cell can be situated in the pre-switched B-cell compartment and that heavy chain switching can occur without further somatic mutation. Based on the MM immunoglobulin sequences derived from the bone marrow, patient-specific CDR2 and CDR3 oligonucleotides were designed. B lymphocytes were separated from plasma cells based on the expression of CD19 and HLA class II or surface bound IgM using immunomagnetic beads. The expressed Ig sequences were amplified by RT-PCR using patient specific CDR2 primers and isotype specific primers (C mu, C gamma, and C alpha). Myeloma-specific Ig sequences were detected by a myeloma-specific CDR3 probe and sequenced. In one out of five cases we found in the peripheral blood clonally related IgM and IgA sequences with the same somatic mutations as the MM-IgG sequence. In another case of an IgG MM we found in the bone marrow clonally related IgA sequences with the same somatic mutations. These findings, together with the fact that myeloma-Ig genes contain somatic mutations without intraclonal variation, suggest that the clonogenic cell in multiple myeloma can originate from a pre-switched but somatically mutated B cell.

Amino Acid Sequence↗