Conservation of the 3'-untranslated regions of calmodulin mRNAs in cetaceans.

Three separate calmodulin (CaM) genes (I, II and III) encoding an identical CaM protein but differing in the 5'- and 3'-untranslated regions of each of the three mRNAs are present and highly conserved in all mammals (so far examined). Primers complementary to the 3'- untranslated region (3'UTR) of each of the three mRNAs occurring in human, rat and mouse were synthesized and used to amplify regions of the 3'UTR from genomic DNA isolated from cetaceans, specifically from the bottled-nosed dolphin (Tursiops truncates), the pygmy sperm whale (Kogia breviceps) and the humpback whale (Megaptera novaeangliae). Using several primers and PCR conditions, the three CaM genes were identified in all three species by this method with one exception. The sequenced regions of the 3'UTRs of the three genes of the cetaceans exhibited a high percentage identity when compared to the corresponding regions of these three CaM mRNAs isolated from humans (85-96%). These partial sequences of the 3'UTR regions and the corresponding regions for humans, rats and mice that were available from the database were aligned and a phylogenetic tree was constructed. The three CaM genes from all species showed a close phylogenetic relationship based on these 3'UTR sequences. Such high conservation of the 3'UTRs suggests a specialized and significant function for this region in mammals.

Sequence homology of the 3'-untranslated region of calmodulin III in mammals.

Calmodulin (CaM) has three separate nonallelic genes that encode for three identical proteins. These genes differ considerably in the 5'- and 3'-untranslated regions (UTR) and in the promoter region. The sequence of the 3'-UTR of CaM III gene for rat and mouse was completed and compared to the human sequence. The rat and mouse 3'-UTR region had an identity of approximately 80% with the human. Three common polyadenylation signals in the 3'-UTR may account for the multiple CaM III transcripts. Although the untranslated regions are distinctively different for the three CaM genes, these regions are highly conserved among mammals. This high sequence conservation suggests an important function in the localization or regulation of CaM mRNA.

Evolutionary aspects of calmodulin.

Calmodulin (CaM) is a major cellular sensor of calcium signaling, interacts with numerous proteins associated with cellular second messenger systems (e.g., cyclic AMP, nitric oxide), and is associated with neurosecretory activity. An identical CaM protein consisting of four helix-loop-helix regions that arose by gene duplication is encoded by three nonallelic mammalian genes that are some of the most highly conserved genes known. Differential tissue and cellular expression of each CaM suggest unique functions that promote strong selective preservation of these replicate, yet distinct, CaM genes in mammals. Each gene displays the same exon-intron arrangement but is characterized by distinct promoter elements and by unique 5'- and 3'-untranslated regions that are highly conserved among human, rat, and mouse. These distinct untranslated regions may permit regulation of CaM levels at discrete cellular sites during differentiation and in highly specialized cell types such as neurons.

Chronic fatigue syndrome and fibromyalgia: clinical assessment and treatment.

Chronic fatigue syndrome (CFS) and fibromyalgia (FM) are closely related illnesses of uncertain etiology. This article reviews the research literature on these biobehavioral conditions, with an emphasis on explanatory models, clinical evaluation of comorbid psychiatric disorders, assessment of stress factors, pharmacologic and alternative therapies, and cognitive-behavioral treatment studies. Furthermore, clinical protocols suitable for professional practice are presented based on an integration of the authors' clinical observations with published data. The article concludes with the recognition that mental health professionals can offer substantial help to these patients.

Calculation and verification of the ages of retroprocessed pseudogenes.

To verify the existence of processed pseudogenes in different primates and their correlation with the estimated age of divergence, selected regions of processed pseudogenes of alpha-enolase, calmodulin II (CALMII), and argininosuccinate synthetase (AS) were amplified by the polymerase chain reaction (PCR) using DNA of blood samples. Published primate divergence times from the accepted paleontological records and the age of the pseudogenes based on molecular clock calculations were compared to data obtained by detection of PCR products exhibiting the expected amplicon size of the pseudogene region. For the alpha-enolase and the CALMII pseudogenes Psi(2), and Psi(3), calculated divergence times were 11, 19, and 36 Myr, respectively. For the AS pseudogenes Psi(1), Psi(3), and Psi(7), the divergence times were calculated to be 21, 25, and 16 Myr, respectively. Primer design and the annealing temperature are critical factors in the detection of pseudogenes in different species and impact greatly on the interpretation of the PCR analysis. The estimated divergence times of the selected pseudogenes utilizing calculations based on the molecular clock theory correlated well with experimental PCR detection of the selected pseudogenes represented in this study.

Symptom patterns in long-duration chronic fatigue syndrome.

OBJECTIVE: Our objective was to evaluate symptom patterns in patients with chronic fatigue syndrome (CFS) who were ill for 10 or more years. METHODS: This cross-sectional self-report study compared patient groups with long-duration (median = 18 years; n = 258) and short-duration (median = 3 years; n = 28) CFS to a group of healthy significant others (n = 79) on symptomatic, neurocognitive, and psychological variables. Data were gathered from a 574-item postal questionnaire. RESULTS: A principal-components analysis of CFS symptom data yielded a three-factor solution: cognitive problems; flu-like symptoms; and neurologic symptoms. Compared with the short-duration CFS group, the long-duration group had significantly higher CFS symptom severity scores (p < 0.04), largely attributable to increased cognitive difficulties. A subgroup comparison of subjects ill for < 3 years versus those ill 4-7 years suggested that denial coping strategies were more likely in those participants with the shorter illness duration. Significant differences between both CFS groups and healthy controls were found in a number of comorbid disorders. Participants with CFS most often endorsed immune/viral abnormalities and persistent stress as important perceived causes of their illness. CONCLUSION: Participants with long-duration CFS reported a large number of specific cognitive difficulties that were greater in severity than those reported by participants with short-duration CFS. The pattern of comorbid disorders in the CFS groups was consistent with hypersensitivity and viral reactivation hypotheses.

Sequence of the human CALM II calmodulin gene promoter region.

A clone containing a portion of the promoter region for the human bona fide CALM II gene was isolated using a human Promoter Finder DNA Walking Kit. This promoter region contains, putatively, a GC box (common in housekeeping genes), a CRE-binding site, a TATA like box and AGGGA sequences. The latter are reported to be present in genes for Ca2+ binding genes. This human promoter region exhibits overall 85% sequence identity to the corresponding region of the rat CALM II promoter but shows no identity to the corresponding region of the human CALM I or CALM III promoters.

Politics, science, and the emergence of a new disease. The case of chronic fatigue syndrome.

Chronic fatigue syndrome (CFS) emerged as a diagnostic category during the last decade. Initial research suggested that CFS was a relatively rare disorder with a high level of psychiatric comorbidity. Many physicians minimized the seriousness of this disorder and also interpreted the syndrome as being equivalent to a psychiatric disorder. These attitudes had negative consequences for the treatment of CFS. By the mid-1990s, findings from more representative epidemiological studies indicated considerably higher CFS prevalence rates. However, the use of the revised CFS case definition might have produced heterogeneous patient groups, possibly including some patients with pure psychiatric disorders. Social scientists have the expertise to more precisely define this syndrome and to develop appropriate and sensitive research strategies for understanding this disease.

Accuracy of indirect estimates of maternal mortality: a simulation model.

A simulation model was developed to test the accuracy of indirect estimates of maternal mortality (the sisterhood method). The model generated a first generation of grandmothers, a second generation of mothers (with brothers and sisters), and a third generation of children (births). In the second generation, maternal mortality was introduced. Empirical values for the parameters of fertility and mortality were taken from a prospective survey in Senegal (Niakhar). Results based on 100 simulations of the same situation revealed several limitations of the sisterhood method: The indirect estimates could fall as far as 33 percent from the true values on individual cases; the indirect estimates tended to be systematically higher than the direct estimates; their range was wider, as were their confidence intervals; and biases were particularly strong for the younger age groups of respondents. Reasons for these biases are explored.

PIP: A simulation method was used to assess the accuracy of indirect estimates of maternal mortality (sisterhood method). The model generated a first generation of grandmothers, a second generation of mothers (with brothers and sisters), and a third generation of children (births). Maternal mortality was introduced in the second generation. Fertility and mortality parameters for the simulations were derived from a prospective study conducted in Niakhar, Senegal, in 1983-89. Results based on 100 simulations of the same situation revealed several limitations of the sisterhood method. The indirect estimates were as far as 33% from the true values on individual cases, despite the assumed perfect quality of the data. The indirect estimates tended to be systematically higher than the direct estimates, with wider ranges and confidence intervals. Biases were especially marked for the younger age group (15-39 years). The only justification for use of indirect rather than direct estimates seems to be the avoidance of questions about age differences between the respondent and the sister and the age of the sister at time of death, when applicable. Recommended, instead of indirect estimates, are two methods of computing direct estimates of maternal mortality: 1) direct computation of the maternal mortality quotient, followed by conversion to the maternal mortality rate through use of the total fertility rate, and 2) use of information on parity by age from maternity histories to compute the maternal mortality rate in each age group directly.

Sequence motifs for calmodulin recognition.

Calmodulin (CaM) is recognized as a major calcium sensor and orchestrator of regulatory events through its interaction with a diverse group of cellular proteins. Many investigations have focused on defining the region of interaction between CaM and its cellular targets and the action of CaM on target protein function. Because CaM can bind with high affinity to a relatively small alpha-helical region of many proteins, success in clearly defining the essential elements of CaM binding motifs seems feasible and should provide a means of identifying CaM binding proteins. Three recognition motifs for CaM interaction are discussed in the context of experimental investigations of a variety of CaM target proteins. A modified version of the IQ motif as a consensus for Ca2+-independent binding and two related motifs for Ca2+-dependent binding, termed 18-14 and 1-5-10 based on the position of conserved hydrophobic residues, are proposed. Although considerable sequence diversity is observed among the different binding regions, these three classes of recognition motifs exist for many of the known CaM binding proteins.

Cognitive functioning and depression in patients with chronic fatigue syndrome and multiple sclerosis.

OBJECTIVE: To assess cognitive function in patients with chronic fatigue syndrome (CFS) and multiple sclerosis (MS) and to evaluate the role of depressive symptoms in cognitive performance. DESIGN: Case-control. All subjects were given a neuropsychological battery, self-report measures of depression and fatigue, and a global cognitive impairment rating by a neuropsychologist "blinded" to clinical diagnosis. Patients with MS and CFS were additionally evaluated with a Structured Clinical Interview for DSM-III-R (Diagnostic and Statistical Manual of Mental Disorders, Revised Third Edition) disorders. SETTING: Institutional and private neurological practices and the community at large. PATIENTS: Twenty patients with CFS diagnosed in accord with the Centers for Disease Control and Prevention-revised criteria who had cognitive complaints; 20 patients with clinically definite MS who were ambulatory and were matched for fatigue severity, age, and education to CFS subjects; and 20 age- and education-matched healthy controls. RESULTS: Patients with CFS had significantly elevated depression symptoms compared with patients with MS and healthy controls (P < .001) and had a greater lifetime prevalence of depression and dysthymia compared with MS subjects. Patients with CFS, relative to controls, performed more poorly on the Digit Symbol subtest (P = .023) and showed a trend for poorer performance on logical memory (P = .087). Patients with MS compared with controls had more widespread differences of greater magnitude on the Digit Span (P < .004) and Digit Symbol (P < .001), Trail Making parts A (P = .022) and B (P = .037), and Controlled Oral Word Association (P = .043) tests. Patients with MS also showed a trend of poorer performance on the Booklet Category Test (P = .089). When patients with CFS and MS were directly compared, MS subjects had lower scores on all measures, but the differences reached significance only for the Digit Span measure of attention (P = .035). CONCLUSIONS: Patients with CFS compared with MS have more depressive symptoms but less cognitive impairment. Relative to controls, a subset of CFS subjects did poorly on tests of visuomotor search and on the logical memory measure of the Wechsler Memory Scale-revised. Poor performance of logical memory in CFS appears to be related to depression, while visuomotor deficits in CFS are unrelated. Cognitive deficits in patients with MS are more widespread compared with those in patients with CFS and are independent of depressive symptoms.

A comparison of cognitive behavioral treatment for chronic fatigue syndrome and primary depression.

To evaluate the effect of cognitive behavioral intervention on chronic fatigue syndrome (CFS), we studied three patient groups: a CFS-treatment group (n = 22), a primary depression-treatment group (n = 20), and a no-treatment control group of subjects with CFS (n = 22). For the CFS-treatment group, a trend toward reduced depression-symptom scores was noted, but there were no significant changes in stress-related symptoms or fatigue severity. For the most depressed treated subjects with CFS, significant score reductions were observed in measures of depression, stress, fatigue severity, and fatigue-related thinking. In the depression group, significant reductions in depression, stress, and fatigue severity scores were found. No significant changes in any measure were observed in the CFS control group. A new fatigue-related cognitions scale, developed to assess cognitive and emotional reactions to fatigue, showed a significant reduction in such reactions in the CFS-treatment group, a finding suggesting that depression in this group was mediated by maladaptive thinking. The results suggest that a subset of CFS patients with cognition-related depressive symptomatology may respond to short-term behavioral intervention.

A comparison of neuropsychiatric characteristics in chronic fatigue syndrome, multiple sclerosis, and major depression.

Chronic fatigue syndrome (CFS), a controversial clinical entity characterized by severe fatigue and constitutional symptoms, has been associated with a variety of psychiatric disorders. To further understand the psychiatric profile of CFS, the authors compared patients with CFS, multiple sclerosis (MS), and major depression by using diagnostic interviews and self-report measures of Axis I disorders and personality disorders. CFS patients differed from patients with major depression, with significantly less depression and fewer personality disorders. Compared with MS patients, CFS patients did not differ with regard to personality disorders. However, they did have significantly more frequent current depression than MS patients, particularly following onset of their illness.

Multiple mRNAs encoding human calmodulin.

In humans, as in rats, four distinct molecular weight species of mRNA encoding calmodulin exist, i.e. 1.6 Kb for L, 1.4 Kb for T2 and 2.5 and 1.0 Kb for T1. They result from the expression of three genes. Each of these mRNAs codes for a calmodulin identical in amino acid sequence. The 5' and 3' untranslated regions of these mRNAs, however, differ extensively, and oligonucleotide probes specific to these regions were used in this study. The poly A+ mRNA was isolated from human erythroleukemia cells and also from human B cells infected with EBV.

Species comparison of calmodulin sequences.

No amino acid substitutions can be located when the calmodulin produced in various vertebrate species (human, rat, chicken, toad) are compared. However, multiple substitutions exist in calmodulin derived from non-vertebrates. Here, we have determined the residues for which no alterations in sequence are allowed. The protein from each species exhibits a sequence identity from residue 27 to residue 53, i.e., residues spanning a small part of the Ca2+ binding loop I and the adjacent interloop region. The analogous sequence (residues 100 to 129) abutting the Ca2+ binding loop III also exhibits only a few differences. Furthermore, negatively charged side chains at residues 82-84 in the central alpha-helix are conserved.

[Results of surgical treatment of vulvar cancer]. / Resultate der operativen Behandlung des Vulvakarzinoms.

129 patients with carcinoma of the vulva were treated at the Dept. of Obstetrics and Gynaecology from 1966-1985. FIGO stage I was observed in 21% of the cases, FIGO stage II in 50%, FIGO stage III in 10% and FIGO stage IV in 8% of the patients. In 10% of the patients, definite classification was not possible. Mean age at the onset of the disease was 66.2 years, the mean time of observation 63 months. Kraurosis of the vulva or leukoplakia were simultaneous phenomena recorded locally in 67% of the patients. Histological investigation showed squamous cell carcinoma in 93% of the cases. The tumours were most frequently observed on the labia and the clitoris. 98% of the patients underwent surgery, 64% radical vulvectomy with inguinal lymph node disection. 56% of the patients of this group had a 5-year survival rate, which was 47% for the entire group of patients. Wound healing disorders were the most frequently observed postoperative complications. Metastasis to the inguinal lymph nodes at the time of diagnosis is the critical point of the prognosis. In the absence of lymph node involvement, 68% of the patients achieved a 5-year survival, in the presence of lymph node involvement, the 5-year survival rate was only 13%. Results obtained by this study support the view, that radical surgery at the earliest possible time is the treatment of choice for carcinoma of the vulva.