Genes and pseudogenes for human U2 RNA. Implications for the mechanism of pseudogene formation.

Three loci, designated U2/4, U2/6 and U2/7, which contain sequences related to human U2 RNA, have been studied. The U2/6 locus contains a tandem array of bona fide U2 genes. U2/4 and U2/7, in contrast, contain pseudogenes of whose sequences deviate significantly from that of mammalian U2 RNA. The two pseudogenes appear to have been created by different mechanisms. The sequences that flank the pseudogene in the U2/4 locus lack homology to the corresponding sequences in functional human U2 genes, except for 10 base-pairs immediately following the 3' end. The conserved 3'-flanking segment is homologous to those nucleotides that are present in a U2 RNA precursor. No direct repeats flank the pseudogene in the U2/4 locus. The observations thus suggest that a complementary DNA copy of the U2 RNA precursor was inserted into a blunt-ended chromosomal break to generate the U2/4 locus. The U2/7 locus, in contrast, revealed flanking sequence homology when compared to functional U2 genes, both on the 5' and 3' sides of the pseudogene. The homology was interrupted on both sides by repetitive sequences belonging to the Alu family. On the 5' side the homology continues beyond the Alu repeats whereas on the 3' side it ends precisely at the Alu repeat. This Alu repeat is inserted in a region where a homocopolymeric region of alternating C and T residues is located in functional U2 loci. The observed organization of the U2/7 locus suggests that a previously functional U2 locus was invaded by Alu repeats and subsequently accumulated base substitutions to become a pseudogene.

Association of susceptibility to multiple sclerosis in Sweden with HLA class II DRB1 and DQB1 alleles.

The association of MS with HLA class II alleles was studied by PCR-based typing of the DQA1, DQB1, DRB1, and DPB1 loci in 94 Swedish patients with relapses and remissions of the disease. The haplotype DRB1*1501-DQA1*0102-DQB1*0602 was found to be positively associated and three haplotypes were found to be negatively associated with MS. Linkage disequilibrium makes it difficult to assess whether DRB1 or DQB1 plays the primary role in the disease association, while the association with DPB1 and DQA1 appears to be secondary to that of DQB1 and DRB1. Two of the three haplotypes negatively associated with MS carry the DQB1*0301 allele. Also, the negatively associated DRB1*0401-DQA1*0301-DQB1*0301 haplotype differs from those with nonassociated DRB1*0401-DQA1*0301-DQB1*0302 haplotype only at DQB1. These results suggest that DQB1 alleles, as well as some DRB1 alleles, are involved in susceptibility and protection to MS. In searching for sequence motifs in the DR beta chain associated with MS susceptibility, all DRB1 alleles on haplotypes positively associated with MS, including the DRB1*1501, were found to encode a Val at position 86 of the DR beta chain. Also, DRB1 alleles that are negatively associated with MS all encode a Gly at position 86, suggesting that the residue at position 86 may be critical in conferring susceptibility and protection to MS. Finally, when the effect of the DRB1*1501 haplotype was removed there was no support for the hypothesis that MS is associated with a putative DQ-alpha beta heterodimer, encoded for by certain DQA1 and DQB1 alleles.

DNA typing of forensic material with mixed genotypes using allele-specific enzymatic amplification (polymerase chain reaction).

Biological material in forensic casework frequently contains a mixture of genotypes, with a predominance of material from the victim and only trace amounts from the person committing the crime. Physical separation of the two genotypes or preferential lysis of different cell types may sometimes be possible. However, it is often difficult to achieve complete separation due to the lysis of cells or lack of material. We have developed an enzymatic amplification system for the HLA DQA1 locus, that will allow the presence of individual alleles in a sample with mixed genotypes to be determined, independent of their initial proportion in the sample. This system permits the identification of an allele representing less than 10(-4) of the background genotype. Use of polymerase chain reaction (PCR) with general primers allows only alleles representing more than about 1% to be detected, while the allele-specific amplification represents up to a 1000-fold increase in sensitivity. This method was applied to a rape case and a combined rape and murder case; in both cases the biological evidential materials contained a mixture of alleles from the victim and the rapist. Allele-specific PCR revealed the presence of alleles identical to those of the suspect using DNA from a vaginal swab taken after a rape incident, whereas by using general primers in the PCR only trace amounts of alleles other than those of the victim were found. Similarly, allele-specific amplification of DNA from vaginal swabs from the murder case revealed the presence of alleles identical to those of the suspect, while standard PCR only indicated the presence of genetic material from the victim.

Renal, thyroid and parathyroid function during lithium treatment: laboratory tests in 207 people treated for 1-30 years.

A total of 207 patients (diagnoses revised according to DSM-III-R) attended our outpatient clinic and were treated with lithium for 1-30 years. They were subjected to conventional renal, thyroid and parathyroid function tests. With increasing treatment duration, the renal tests showed only moderate deviations from expected reference values. No patient developed renal insufficiency. Oversubstitution (thyrotropin < or = 0.1 mU/l) was suspected in 25% of the patients on thyroxine. Cross-sectionally unrecognized hypothyroidism was found in 6% of the patients. Elevated ionized serum calcium was found in 25% and elevated serum intact parathyroid hormone in 23% of the patients.

The effect of lithium treatment on manic symptoms and levels of monoamine metabolites in cerebrospinal fluid of manic depressive patients.

Clinical effects, levels of 5-hydroxyindoleacetic acid (5-HIAA) and homovanillic acid (HVA) in cerebrospinal fluid (CSF) and lithium levels in serum were examined in 13 manic depressive patients acutely admitted because of a manic or hypomanic episode. Patients were examined before and 12 days after the beginning of lithium treatment. Manic scores were significantly reduced during treatment. The levels of 5-HIAA as well as HVA increased significantly during treatment. The HVA to 5-HIAA ratio was significantly reduced, indicating a more pronounced change in 5-HIAA than in HVA. The 5-HIAA and HVA levels before as well as after 12 days of treatment were significantly correlated. No significant correlation was found between manic scores and monoamine metabolites in CSF or between lithium level in serum and reduction of manic scores or elevation of monoamine metabolites in CSF in the relative small number of patients studied.

Triglyceride turnover, lipoprotein lipase activity, and fat cell size in adipose tissue of rats during the first 2 weeks of pregnancy.

Understanding the biological function of the fat retained during pregnancy is important when estimating energy needs during reproduction. Women as well as rats are often considered to gain fat at specific anatomical sites during pregnancy and use this fat as a source of energy during lactation. However, mobilized body fat covers only a minor part of the energy needed by the lactating rat dam. In this paper, fat cell size, lipoprotein lipase activity, as well as triglyceride turnover in parametrial and subcutaneous adipose tissues were studied during the first 2 weeks of gestation and in virginal controls to further explore the metabolic and physiological basis for changes in body fat during reproduction in rats. Pregnancy increased the size of parametrial but not of subcutaneous adipocytes. The lipoprotein lipase activity in subcutaneous adipocytes was not increased by pregnancy. The accumulation in adipose tissue of 14C from orally administered 14C-oleic acid was higher in virginal than in pregnant rats. No effect of pregnancy on the rate of lipid turnover in parametrial or subcutaneous adipocytes was found. The findings are in accordance with the contention that body fat gained during rat pregnancy, to a large extent, is a consequence of a general growth of maternal tissues rather than the result of a stimulating effect of pregnancy on fat accumulation by adipocytes from specific body sites.

Schizoaffective psychoses. A long-term follow-up.

The study comprises 20 men who had shown admixtures of schizophrenic and affective symptoms during their first hospitalisation in a psychiatric hospital in Stockholm 1946-1957. The observation period was between 17 and 30 years. At the time of follow-up the group was rediagnosed on the basis of all information obtained. It was shown that this originally relatively homogeneous group was made up of a heterogeneous collection of illnesses. Definite schizophrenia formed the largest group (nine patients). Three patients had developed a definite manic-depressive disease and three severe alcoholism. The remaining five patients showed prominent symptoms of both schizophrenic and affective type. After the first hospitalisation 19 patients were recovered or improved. At the end of the follow-up period the corresponding number was eight. Twelve patients showed social recovery, i.e. were working or studying at the end of the follow-up period. Eighteen patients were at some time re-hospitalised for psychosis. The group, as a whole, spent 16% of the observation period hospitalised in a mental hospital - 6% in the outcome group recovered + improved and 21% in the outcome group were unimproved. The course was difficult to predict with the aid of factors usually considered prognostically favourable. The study stresses the need for prospective controlled studies based on a systematised after-care programme for patients who show symptoms of both schizophrenia and affective illness in their first attack.

Mortality in 497 patients with affective disorders attending a lithium clinic or after having left it.

The impact of lithium prophylaxis on mortality has been studied in 497 patients, 405 bipolars and 92 unipolars, who attended the same out-patient lithium clinic for up to 30 years. In order to avoid preselection, no minimum period of lithium treatment was required in our study. Of a total of 6014 patient-years, 4330 were spent in regular contact with the study clinic. General mortality due to natural causes was not significantly increased; among cardiovascular diseases, only pulmonary embolism showed an excess mortality. No patients died of lithium intoxication or chronic renal insufficiency. Patients were divided into three groups: Group A, 277 patients, attended the study clinic until death or the end of the study, Group B, 86 patients, left the clinic but continued to take lithium, and Group C, 134 patients, both left the clinic and stopped taking lithium. Among bipolars, the suicide rate compared to the general population was in excess in all three groups. Among unipolars, suicides occurred only after the patients had left the study clinic and stopped taking lithium. A special analytical method was used for intergroup comparisons of suicide rates. Bipolars in Group A attending the study clinic regularly had a suicide rate of 3.5 per 1000 patient-years. The rate increased to 6.3 or by 80 % if patients had left the clinic and did not take lithium any longer as in Group C. The suicide rate in Group C increased by 45% compared to Group B, patients who left the clinic but continued to take lithium. Our results support the hypothesis that lithium has a significant antisuicidal effect in bipolars as well as in unipolars. The suicide mortality can be further reduced by regular attendance in a specialised mood disorder clinic.

Complex genetic control in a rat model for rheumatoid arthritis.

We have defined previously five quantitative trait loci controlling development of pristane-induced arthritis in a cross between E3 and DA rats. To define new loci controlling the disease we have mapped three recombinant inbred strains between DA and E3 and analysed an F2 cross between DA rats and one of these RI strains. Two novel loci affecting disease severity are identified on chromosome 1 (Pia8) and chromosome 4 (Pia7) respectively. We could also reproduce the earlier identified Pia3 locus on chromosome 6 associated with arthritis onset. In the original E3xDA F2 cross, neither of the loci Pia7, Pia8, or Pia1 showed any association with arthritis. To investigate the possibility of interacting loci preventing the phenotypic expression of other loci, the E3xDA F2 cross was re-analysed with a model for a two locus interaction, knowing the presence of these newly identified loci. We found suggestive evidence for an interaction where an effect of Pia7 and Pia1 on disease severity depends on DA homozygosity at specific loci, which themselves do not confer susceptibility. This shows that additional disease associated loci can be identified if other loci are neutralized. This will be of importance for understanding the complex genetics controlling rheumatoid arthritis.

Group therapy with schizophrenic patients in outpatient departments.

Twelve schizophrenic patients were treated with neuroleptic drugs and psychoanalytically oriented group therapy during a period of 2 years. Twelve other patients, matched with regard to the state of their disease, sex, age, civil status and social situation, were given neuroleptic drugs and contact therapy during the same period. All patients were evaluated by the same test procedures. The Rorschach test, the Defence Mechanism Test (DMT), interviews and a self-evaluation test, were performed before and after 2 years of treatment. The dates of discharge, number of days in hospital and neuroleptic drugs prescribed were recorded for all patients over a 2-year period before, during and after treatment. Half of the patients improved, regardless of the treatment they received. No evaluation instrument used before the start of treatment could predict the patients who later improved. After 2 years of treatment, it was assessed that the patients who improved required a further period of insight therapy.

HLA DQ-DR haplotype and susceptibility to cervical carcinoma: indications of increased risk for development of cervical carcinoma in individuals infected with HPV 18.

The association of HLA class II DQB1 and DRB1 alleles with the development of cervical carcinoma was studied in 150 Swedish patients using PCR-based HPV and HLA typing. The association of cervical carcinoma with alleles encoding the DQ3 antigen, previously found among German and Norwegian patients, was not observed in the Swedish patients. Five DQ-DR haplotypes were indicated to be positively associated with development of cervical carcinoma in the Swedish patients. Two of these HLA associations were specific for HPV 18 infected patients, suggesting that the ability of the oncogenic HPV 18 to cause more rapid-transit tumors than other high risk HPV types may be due to a deficiency in antigen presentation by the HLA molecules encoded by carried on these haplotypes.

Repeated application of EMLA cream 5% for the alleviation of cannulation pain in haemodialysis.

The analgesic effect and the occurrence of local reactions after repeated application of a lidocaine/prilocaine cream (EMLA 5%) were investigated in 31 haemodialysis patients. The cream was used for alleviation of cannulation pain prior to the haemodialysis (HD) for a period of 1-1.5 years. In each patient 300 to 312 applications of the cream were made during the period. Local skin reactions were recorded after the cream applications and the analgesic effect was evaluated with double-blind, crossover placebo controls at regular intervals. Seventeen patients completed the study. EMLA gave considerable pain relief and was significantly better than placebo at all effect evaluations but one. The frequency of local reactions was low and not correlated to the number of applications of EMLA. Two patients, however, interrupted their treatment due to local irritation.