Trends in thyroid function testing, neck ultrasound, thyroid fine needle aspiration, and thyroidectomies in North-eastern Italy.

PURPOSE: Evidence of an increased diagnostic pressure on thyroid has emerged over the past decades. This study aimed to provide estimates of a wide spectrum of surveillance indicators for thyroid dysfunctions and diseases in Italy. METHODS: A population-based study was conducted in North-eastern Italy, including 11.7 million residents (20% of the total Italian population). Prescriptions for TSH testing, neck ultrasound or thyroid fine needle aspiration (FNA), surgical procedures, and drugs for hypo- or hyperthyroidism were extracted from regional health databases. Proportions and rates of selected examinations were calculated from 2010 to 2017, overall and by sex, calendar years, age, and region. RESULTS: Between 2010 and 2017 in North-eastern Italy, 24.5% of women and 9.8% of men received at least one TSH test yearly. In 2017, 7.1% of women and 1.5% of men were prescribed drugs for thyroid dysfunction, 94.6% of whom for hypothyroidism. Neck ultrasound examinations were performed yearly in 6.9% of women and 4.6% of men, with a nearly two-fold variation between areas. Thyroid FNA and thyroidectomies were three-fold more frequent in women (394 and 85 per 100,000) than in men (128 and 29 per 100,000) with a marked variation between areas. Both procedures decreased consistently after 2013. CONCLUSIONS: The results of this population-based study describe recent variations over time and between surrounding areas of indicators of 'diagnostic pressure' on thyroid in North-eastern Italy. These results emphasize the need to harmonize practices and to reduce some procedures (e.g., neck ultrasound and total thyroidectomies) in certain areas.

A novel mutation in NDUFB11 unveils a new clinical phenotype associated with lactic acidosis and sideroblastic anemia.

NDUFB11, a component of mitochondrial complex I, is a relatively small integral membrane protein, belonging to the "supernumerary" group of subunits, but proved to be absolutely essential for the assembly of an active complex I. Mutations in the X-linked nuclear-encoded NDUFB11 gene have recently been discovered in association with two distinct phenotypes, i.e. microphthalmia with linear skin defects and histiocytoid cardiomyopathy. We report on a male with complex I deficiency, caused by a de novo mutation in NDUFB11 and displaying early-onset sideroblastic anemia as the unique feature. This is the third report that describes a mutation in NDUFB11, but all are associated with a different phenotype. Our results further expand the molecular spectrum and associated clinical phenotype of NDUFB11 defects.

Long-term survival, prevalence, and cure of cancer: a population-based estimation for 818 902 Italian patients and 26 cancer types.

BACKGROUND: Persons living after a cancer diagnosis represent 4% of the whole population in high-income countries. The aim of the study was to provide estimates of indicators of long-term survival and cure for 26 cancer types, presently lacking. PATIENTS AND METHODS: Data on 818 902 Italian cancer patients diagnosed at age 15-74 years in 1985-2005 were included. Proportions of patients with the same death rates of the general population (cure fractions) and those of prevalent patients who were not at risk of dying as a result of cancer (cure prevalence) were calculated, using validated mixture cure models, by cancer type, sex, and age group. We also estimated complete prevalence, conditional relative survival (CRS), time to reach 5- and 10-year CRS >95%, and proportion of patients living longer than those thresholds. RESULTS: The cure fractions ranged from >90% for patients aged <45 years with thyroid and testis cancers to <10% for liver and pancreatic cancers of all ages. Five- or 10-year CRS >95% were both reached in <10 years by patients with cancers of the stomach, colon-rectum, pancreas, corpus and cervix uteri, brain, and Hodgkin lymphoma. For breast cancer patients, 5- and 10-year CRSs reached >95% after 19 and 25 years, respectively, and in 15 and 18 years for prostate cancer patients. Five-year CRS remained <95% for >25 years after cancer diagnosis in patients with liver and larynx cancers, non-Hodgkin lymphoma, myeloma, and leukaemia. Overall, the cure prevalence was 67% for men and 77% for women. Therefore, 21% of male and 31% of female patients had already reached 5-year CRS >95%, whereas 18% and 25% had reached 10-year CRS >95%. CONCLUSIONS: A quarter of Italian cancer patients can be considered cured. This observation has a high potential impact on health planning, clinical practice, and patients' perspective.

Oesophageal cancer survival in Europe: a EUROCARE-4 study.

Oesophageal cancer survival is poor with variation across Europe. No pan-European studies of survival differences by oesophageal cancer subtype exist. This study investigates rates and trends in oesophageal cancer survival across Europe. Data for primary malignant oesophageal cancer diagnosed in 1995-1999 and followed up to the end of 2003 was obtained from 66 cancer registries in 24 European countries. Relative survival was calculated using the Hakulinen approach. Staging data were available from 19 registries. Survival by region, gender, age, morphology and stage was investigated. Cohort analysis and the period approach were applied to investigate survival trends from 1988 to 2002 for 31 registries in 17 countries. In total 51,499 cases of oesophageal cancer diagnosed 1995-1999 were analysed. Overall, European 1- and 5-year survival rates were 33.4% (95% CI 32.9-33.9%) and 9.8% (95% CI 9.4-10.1%), respectively. Males, older patients and patients with late stage disease had poorer 1- and 5-year relative survival. Patients with squamous cell carcinoma had poorer 1-year relative survival. Regional variation in survival was observed with Central Europe above and Eastern Europe below the European pool. Survival for distant stage disease was similar across Europe while survival rates for localised disease were below the European pool in Eastern and Southern Europe. Improvement in European 1-year relative survival was reported (p=0.016). Oesophageal cancer survival was poor across Europe. Persistent regional variations in 1-year survival point to a need for a high resolution study of diagnostic and treatment practices of oesophageal cancer.

Prognoses for head and neck cancers in Europe diagnosed in 1995-1999: a population-based study.

BACKGROUND: head and neck cancers are a heterogeneous group of malignancies, affecting various sites and subsites, with differing prognoses. The aim of this study was to analyse survival for European head and neck cancer patients in populations covered by population-based cancer registries (CRs), in relation to tumour subsite as prognostic factor. PATIENTS AND METHODS: we analysed 51 912 adult head and neck cancer cases (36 322 mouth-pharynx and 15 590 larynx) diagnosed from 1995 to 1999 and archived by 45 CRs in 20 countries participating in EUROCARE-4. Five-year age-standardised relative survival was estimated for mouth-pharynx and larynx sites by sex and country. Relative survival was modelled to provide estimates of relative excess risks (RERs) of death by country, adjusted for confounding factors. RESULTS: a large but site-variable proportion of tumours were incompletely specified. Five-year age-standardised relative survival was low in Slovakia and high in The Netherlands. Adjustment for subsite reduced RERs of death for most countries; 5-year relative survival increased from 1990-1994 to 1995-1999 for all subsites, while between-country differences in survival narrowed. CONCLUSION: differences in subsite distribution explain a considerable part of the survival differences for head and neck cancers, however, incomplete/inaccurate subsite reporting complicate interpretation.

Estimating regional cancer burden in countries with partial registration coverage: an application to all malignant neoplasms in Italy over the period 1970-2010.

Regional epidemiological indicators of cancer burden are essential information for cancer surveillance and health resources planning, especially in countries with partial registration coverage and geographically variable risk patterns, such as Italy. This paper presents a methodology to derive cancer incidence and prevalence at the regional and national scale and illustrates its application to all malignant neoplasms in Italy for the period 1970-2010. The method, denoted as MIAMOD, is based on a back-calculation approach and derives cancer-specific morbidity measures by using official mortality data and model-based relative survival from local Cancer Registries data. The output includes time-trends and projections of a complete set of epidemiological indicators, i.e. mortality, incidence and prevalence. Results for all cancers in Italy show different incidence patterns by gender and a pronounced regional variability among men: male incidence is estimated to decrease in almost all northern-central regions, while more stable or even rising trends are estimated in the southern regions. No incidence reduction is expected for women. Prevalence increases country-wide in both sexes. The proposed approach can be applied to derive regional up-to-date time trends of cancer burden indicators in countries with local and sparse cancer registration systems. These estimates are useful for planning health services on a national and regional basis and for highlighting regional differences.

Mutations in yeast mt tRNAs: specific and general suppression by nuclear encoded tRNA interactors.

Mutations in mitochondrial tRNA genes can produce alterations in tRNA structure resulting in defective mitochondrial protein synthesis and hence respiratory defects. Such defects are often at the origin of neurodegenerative diseases in humans and can be easily studied in yeast since respiratory deficient mutants are viable. Several nuclear encoded tRNA interactors have been shown to rescue the mitochondrial defects due to mutations in mitochondrial tRNAs. Among these, we have identified the gene for the mitochondrial protein synthesis elongation factor EF-Tu and the specific mt aminoacyl-tRNA synthetases. We also observed that the respiratory defects and the effect of the TUF1 over-expression were strongly strain dependent. The importance of the nuclear background in which the mitochondrial mutation is expressed was investigated by changing the nuclear context. Finally, we demonstrated, using the RT-PCR method, the existence of significantly variable levels of the TUF1 transcript among strains with functional and dysfunctional mitochondria.

Nucleo-mitochondrial interactions in Saccharomyces cerevisiae: characterization of a nuclear gene suppressing a defect in mitochondrial tRNA(Asp) processing.

We utilized the heat-sensitive mutant strain (Ts932), bearing a mutation at position 61 in the mitochondrial tRNA(Asp) gene, to identify nuclear genes involved in tRNA biogenesis; this mutant is defective in 3'-end processing and consequently in the production of mature mitochondrial tRNA(Asp). We transformed this strain with a yeast nuclear library and we isolated among other suppressors, an unknown, non-essential gene (called SMM1, corresponding to open reading frame YNR015w), which restored the growth on glycerol and a normal amount of processed tRNA(Asp) in the mutant. The gene contains a domain highly conserved in evolution from bacteria to human and its product has been recently shown to have dihydrouridine synthase activity.

Cancer prevalence in Central Europe: the EUROPREVAL Study.

BACKGROUND: Information on cancer prevalence is either absent or largely unavailable for central European countries. MATERIALS AND METHODS: Austria, Germany, The Netherlands, Poland, Slovakia, Slovenia and Switzerland cover a population of 13 million inhabitants. Cancer registries in these countries supplied incidence and survival data for 465 000 cases of cancer. The prevalence of stomach, colon, rectum, lung, breast, cervix uteri, corpus uteri and prostate cancer, as well as skin melanoma, Hodgkin's disease, leukaemia and all malignant neoplasms combined was estimated for the end of 1992. RESULTS: A large heterogeneity was observed within central European countries. For all cancers combined, estimates ranged from 730 per 100 000 in Poland (men) to 3350 per 100 000 in Germany (women). Overall cancer prevalence was the highest in Germany and Switzerland, and the lowest in Poland and Slovenia. In Slovakia, prevalence was higher than average for men and lower than average for women. This was observed for almost all ages. As shown by incidence data, breast cancer was the most frequent malignancy among women in all countries. Among men, prostate cancer was the leading malignancy in Germany, Austria and Switzerland, and lung cancer was the major cancer in Slovenia, Slovakia and Poland. The Netherlands had a high prevalence of both prostate and lung cancer. Time-related magnitude of prevalence within each country and the variability of such proportions across the countries has been estimated and cancer prevalence is given by time since diagnosis (1 year, 1-5 years, 5-10 years, >10 years) for each site. The weight of 1-year prevalence (248 per 100 000 among men and 253 per 100 000 among women) was <15% of total prevalence. Prevalent cases between 1 and 5 years since diagnosis represented between 22% and 34% of the total prevalence. Prevalent cases diagnosed from 5 to 10 years before (335 per 100 000 for men and 505 per 100 000 for women) represented between 17% and 23% of prevalent cancers. Finally, long-term cancer prevalence (diagnosed >10 years before), reflecting long-term survival, and number of people considered as cured from cancer were 490 per 100 000 for men and 1028 per 100 000 for women, with a range between 26% (The Netherlands, men) and 50% (Slovakia, women). CONCLUSION: It is clear from observing countries in Central Europe, that high cancer prevalence is associated with well-developed economies. This burden of cancer could be interpreted as a paradoxical effect of better treatments and thereby survival. It could also be taken as a sign for not being satisfied with the advances in treating patients diagnosed with cancer, and for supporting more primary prevention.

The yeast counterparts of human 'MELAS' mutations cause mitochondrial dysfunction that can be rescued by overexpression of the mitochondrial translation factor EF-Tu.

We have taken advantage of the similarity between human and yeast (Saccharomyces cerevisiae) mitochondrial tRNA(Leu)(UUR), and of the possibility of transforming yeast mitochondria, to construct yeast mitochondrial mutations in the gene encoding tRNA(Leu)(UUR) equivalent to the human A3243G, C3256T and T3291C mutations that have been found in patients with the neurodegenerative disease MELAS (for mitochondrial 'myopathy, encephalopathy, lactic acidosis and stroke-like episodes'). The resulting yeast cells (bearing the equivalent mutations A14G, C26T and T69C) were defective for growth on respiratory substrates, exhibited an abnormal mitochondrial morphology, and accumulated mitochondrial DNA deletions at a very high rate, a trait characteristic of severe mitochondrial defects in protein synthesis. This effect was specific at least in the pathogenic mutation T69C, because when we introduced A or G instead of C, the respiratory defect was absent or very mild. All defective phenotypes returned to normal when the mutant cells were transformed by multicopy plasmids carrying the gene encoding the mitochondrial elongation factor EF-Tu. The ability to create and analyse such mutated strains and to select correcting genes should make yeast a good model for the study of tRNAs and their interacting partners and a practical tool for the study of pathological mutations and of tRNA sequence polymorphisms.

Measuring cancer prevalence in Europe: the EUROPREVAL project.

Cancer prevalence is the proportion of individuals in a population who at some stage during their lifetime have been diagnosed with cancer, irrespective of the date of diagnosis. Cancer prevalence statistics have generally been provided by a limited number of well established cancer registries that have been in existence for several decades. The advent of systematic follow-up of life status of incident cases and the availability of new statistical methodologies, now makes it possible for registries established during the 1970s or 1980s to provide prevalence data. The main problems encountered in the estimation of prevalence are the inclusion of: (i) cases lost to follow-up; (ii) cases known only from their death certificate; (iii) cases diagnosed before the start of registration; and (iv) the treatment of multiple tumours and migrations. The main aim of this paper was to review these problems and discuss, through the experience gained with EUROPREVAL, how they can be overcome. A method is presented for the calculation of prevalence of all cancers combined in the populations covered by the 45 cancer registries participating in EUROPREVAL. Prevalence of cancer is estimated to be 2% on average, with the highest values (3%) in Sweden and the lowest in Eastern Europe, with a minimum of approximately 1% in Poland.

Reintroduction of a characterized Mit tRNA glycine mutation into yeast mitochondria provides a new tool for the study of human neurodegenerative diseases.

We report the identification and characterization of a new mutation (ts9) in the Saccharomyces cerevisiae mitochondrial genome, which was first genetically mapped in the tRNAgly region and further identified by means of sequencing as consisting of a G to A transition at position 30 in the tRNA. The mutation causes an almost complete disappearance of mature tRNAgly, while a second mitochondrial mutation with a compensatory C to T change restores it in normal quantities; this points to the importance of the strong bond between bases 30 and 40 of the anticodon stem in the stabilization of the tRNA. In addition to resulting in a clear-cut heat-sensitive phenotype, the ts9 mutation creates a new EcoRV restriction site. Both properties were used as markers to monitor the successful (re) introduction of the mutated allele into a wild-type mitochondrial genome through biolistic transformation. The mutant frequency in the progeny as well as the correct integration of the mutated allele at its proper site demonstrate the feasibility of this method for creating and investigating specific mitochondrial tRNA mutations. The method will provide important applications for the use of yeast as a model system of human mitochondrial pathologies.

Disruption of six novel genes from chromosome VII of Saccharomyces cerevisiae reveals one essential gene and one gene which affects the growth rate.

Six ORFs of unknown function located on chromosome VII of Saccharomyces cerevisiae were disrupted in two different genetic backgrounds, and the phenotype of the generated mutants was analysed. Disruptions of ORFs YGR256w, YGR272c, YGR273c, YGR275w and YGR276c were carried out using the disruption marker kanMX4 flanked by short homology regions, whereas ORF YGR255c was inactivated with a long flanking homology (LFH) disruption cassette (Wach et al., 1994). Tetrad analysis of the heterozygous disruptants revealed that ORF YGR255c, previously identified as COQ6 and encoding a protein involved in the biosynthesis of coenzime Q (Tzagoloff and Dieckmann, 1990), is an essential gene. The same analysis also revealed that sporulation of the ygr272cDelta heterozygous diploid produced two small colonies per ascus that were also G418-resistant, indicating that the inactivation of ORF YGR272c could result in a slower growth rate. This result was confirmed by growth tests of the haploid disruptants and by complementation of the phenotype after transformation with a plasmid carrying the cognate gene. No phenotypes could be associated to the inactivation of ORFs YGR256w, YGR273c, YGR275w and YGR276c. Two of these genes have recently been further characterized: ORF YGR255w, renamed RTT102, encodes a regulator of the Ty1-element transposition, whereas ORF YGR276c was found to encode the 70 kDa RNase H activity and was renamed RNH70 (Frank et al., 1999).

Cancer prevalence in Italian cancer registry areas: the ITAPREVAL study. ITAPREVAL Working Group.

AIM: To present data on cancer prevalence for the areas covered by Italian cancer registries, by using a standardized set of data collection and elaboration criteria, and a single method of data analysis. SUBJECTS AND METHODS: Data on over 250,000 patients with cancer, diagnosed between 1978 and 1992, from 11 Italian cancer registries covering about 12% of the Italian population were collected, validated and analyzed according to the unified protocol of the ITAPREVAL project. The method implemented in the PREVAL computer program was used to provide prevalence estimates for the period covered by cancer registration. The total prevalence for each registry and for the pool of all registries was then estimated by correcting for incomplete observations due to the period in which the registration was not yet activated. All prevalence estimates were for 1992. RESULTS: Prevalence figures are presented by cancer site, age, sex, years from diagnosis and registry area. For all malignancies combined, total prevalence ranged from 1,350 per 100,000 inhabitants in Ragusa to 3,650 per 100,000 inhabitants in Romagna, the ratio between these two extremes being 2.7. For the pool of the areas covered by registration cancer prevalence was 3,100 per 100,000 females and 2,250 per 100,000 males. About a third of the total female cases and about half the male cases were diagnosed in the previous five years. Among those aged over 75 years, total prevalence was higher for males than for females: 11,300 versus 8,900 per 100,000 respectively. CONCLUSIONS: This is the first large-scale estimate of the burden of cancer in Italy. It is also one of the first studies in the world which was aimed to study cancer prevalence in detail. These data are necessary for predicting health service needs and help in the evaluation of differences in health service demand by sex, age and Italian regions.

The prevalence of colorectal cancer in Italy.

AIMS: To analyze the prevalence of colorectal cancer (CRC) in different areas of Italy by age, interval since diagnosis and disease stage at diagnosis, and to estimate the prevalence of CRC. These data provide estimates of patient demand on health resources. PATIENTS AND METHODS: Eleven Italian cancer registries (CRs) provided data on 33,740 patients observed for up to 15 years. For the 1,829 cases from the specialized colorectal cancer registry of Modena we analyzed prevalence by Dukes' stage and family history. PREVAL software produced observed prevalence figures by time from diagnosis; to determine the total prevalence, correction factors were applied to the observed data. RESULTS: At the end of 1992, five-year CRC prevalence was high (close to 200 per 100,000) in Genova, Parma, Romagna and Firenze, and low (around 75 per 100,000) in the southern areas of Latina and Ragusa. For all CRs, 86 patients per 100,000 population were alive up to 2 years from diagnosis and 77 per 100,000 between 2 and 5 years from diagnosis. The 5-year prevalence of patients diagnosed with Dukes' B or C (high risk of recurrence and requiring postoperative surveillance) was 152 per 100,000; that of Dukes' A patients 36 per 100,000 (considered cured after surgery and not requiring intensive follow-up or care); that of unstaged patients plus those with distant metastasis at diagnosis was 28 per 100,000 at 5 years (requiring palliative care but not follow-up). The 12-year prevalence of HNPCC was 23 per 100,000, or about 7% of the total; for such patients knowledge of the long-term prevalence is important because they are diagnosed young and are at high risk of multiple tumor development. CONCLUSIONS: 70% of the prevalent patients diagnosed within 5 years prior to the prevalence date were likely to require care for cancer recurrence, while 13% of the prevalent cases required care for distant metastases.

The prevalence of tumors of the breast and female genital tract in Italy.

Data from 10 Italian population-based cancer registries were used to estimate the prevalence of female tumors of the breast and genital tract. The total prevalence, expressed in number per 100,000, was highest for breast cancer (1,117), followed by cancer of the corpus (264) and cervix uteri (146), ovary (110), and vagina and vulva (23). For all tumors the prevalence increased with age at diagnosis. The cancer prevalence was divided into intervals from diagnosis, expressing different health needs in terms of therapy and intensity of clinical follow-up. For all tumors considered, 1-year prevalence was higher than 1-2-year prevalence, reflecting a high death risk due to perioperative mortality and to the proportion of patients diagnosed at advanced stages. The prevalence decreased in the following intervals considered. Noticeable geographic variability was observed in the prevalence across Italy, with higher proportions being registered in the northern-central regions than in the South. The two extreme 0-5-year prevalence figures (per 100,000) were: for breast cancer 568 (Genova) and 259 (Ragusa); for corpus uteri cancer 94 (Romagna) and 21 (Latina); for cervix uteri cancer 63 (Romagna) and 26 (Latina); for ovarian cancer 49 (Parma) and 21 (Latina); for cancer of the vagina and vulva 17 (Genova) and 5 (Ragusa). This variability depends mainly on incidence and on the proportion of elderly in the general population. From 1987 to 1992 there was an increase in the prevalence of tumors of the breast, ovary and vagina and vulva, especially in the elderly. The prevalence of corpus uteri cancer decreased slightly in the elderly only, whereas that of cervix uteri cancer decreased at all ages.

Ts mutations in mitochondrial tRNA genes: characterization and effects of two point mutations in the mitochondrial gene for tRNAphe in Saccharomyces cerevisiae.

Two new mitochondrial mutations conferring heat sensitivity on glycerol medium to the cells that carry them and affecting mitochondrial protein synthesis were investigated. Both map in the mitochondrial tRNAphe gene and have C-to-U transitions, one at position 2 (ts22b16) and the other at 62 (ts1345). The latter mutation clearly affects the 3' end-maturation of tRNAphe, while the former presents normal patterns of both tRNA processing and amino-acylation. The defective phenotype resulting from the ts22b16 mutation can be corrected by over-expressing either the mitochondrial elongation factor EF-Tu or the mutated form of the tRNA. These results suggest that this mutation's primary effect might involve modified interactions during the ternary complex formation.

Suppression of a mitochondrial point mutation in a tRNA gene can cast light on the mechanisms of 3' end-processing.

We used a genetic approach to study the nuclear factors involved in the biogenesis of mitochondrial tRNAs. A point mutation in the mitochondrial tRNA(Asp) gene of Saccharomyces cerevisiae had previously been shown to result in a temperature-sensitive respiratory-deficient phenotype as a result of the absence of 3' end-processing of the tRNA(Asp). Analysis of mitochondrial revertants has shown that all revertants sequenced have a G-A compensatory change at position 53, which restores the hydrogen-bond with the mutated nucleotide. We then searched for nuclear suppressors to identify the nuclear gene(s) involved in mitochondrial tRNA 3' end-processing. One such suppressor mutation was further characterized: it restores tRNA(Asp) maturation and growth at 36 degrees C on glycerol medium in heterozygous diploids, but leads to a defective growth phenotype in haploids.

Symmetrical transcription in the tRNA region of the mitochondrial genome of Saccharomyces cerevisiae.

The occurrence of discrete transcripts originating from the non-coding strand of the yeast mitochondrial genome is described. The region under investigation is localized in the large tRNA gene cluster between the LSU ribosomal RNA and OXI 1 genes. The transcripts originating from the non-coding strand were detected in a wild-type strain and in a rho- mutant. Their size range includes transcripts of about 2000 nucleotides able to accommodate more than one "anti-tRNA". In some cases their extremities can be mapped near highly-conserved nonanucleotides that could function as origins of transcription. The involvement of the tRNA-processing machinery in the cleavage of these transcripts is also hypothesized.

A point mutation in a mitochondrial tRNA gene abolishes its 3' end processing.

A temperature sensitive mutation mapping in the tRNA region of the mitochondrial genome of S. cerevisiae has been found to abolish 3' processing of tRNA(asp). Mutant cells grown for a few generations at the non-permissive temperature were found to specifically lack mature tRNA(asp) and to accumulate 3' unprocessed precursors of this tRNA. The accumulation of precursors of other mitochondrial tRNAs was also observed under the same conditions. After longer incubation times, a generalized decrease of mitochondrial transcripts could be observed. The mutation was genetically mapped in a limited region surrounding the tRNA(asp) gene and found, by sequencing, to consist of a C- greater than T transition at position 61 of the tRNA(asp) gene.