Cytohistological correlation and discrepancy of conventional Papanicolaou smear test with corresponding histopathology: a retrospective study over a 5-year period.

The accuracy of Papanicolaou (Pap) smear testing can be measured using the cytohistological correlation and discrepancy method. This study aimed to evaluate and compare the cytohistological correlation and discrepancy of the conventional Pap test with the corresponding histopathology and to compare the data with other similar studies. A retrospective study was performed at the pathology department of a referral hospital in Muscat, Oman, over a 5-year period. Of 6000 Pap smears, 162 had matching histopathology results (abnormal smear rate 2.7%) but 10 were unsatisfactory for histological diagnosis. Cytohistological correlation was seen in 96/152 (63.2%), while discrepancy was seen in 56/152 (36.8%). False negatives and false positives were found for 7 and 49 cases respectively. The findings of this study confirm the role of conventional Pap testing as a screening test for the diagnosis of cervical lesion but not for management of patients. In comparison with other studies, we also report a low percentage of abnormal Pap smears.

Cytohistological correlation and discrepancy of conventional Papanicolaou smear test with corresponding histopathology:a retrospective study over a 5-year period

The accuracy of Papanicolaou [Pap] smear testing can be measured using the cytohistological correlation and discrepancy method. This study aimed to evaluate and compare the cytohistological correlation and discrepancy of the conventional Pap test with the corresponding histopathology and to compare the data with other similar studies. A retrospective study was performed at the pathology department of a referral hospital in Muscat, Oman, over a 5-year period. Of 6000 Pap smears, 162 had matching histopathology results [abnormal smear rate 2.7%] but 10 were unsatisfactory for histological diagnosis. Cytohistological correlation was seen in 96/152 [63.2%], while discrepancy was seen in 56/152 [36.8%]. False negatives and false positives were found for 7 and 49 cases respectively. The findings of this study confirm the role of conventional Pap testing as a screening test for the diagnosis of cervical lesion but not for management of patients.In comparison with other studies, we also report a low percentage of abnormal Pap smears

L'exactitude du test de Papanicolaou classique peut être mesurée à l'aide de la méthode de la corrélation et de la discordance cytohistologiques. La présente étude visait à évaluer et à comparer la corrélation et la discordance cythohistologiques du test de Papanicolaou classique à l'histopathologie correspondante et à comparer les données à celles d'autres études similaires. Une étude rétrospective a été menée dans le service de pathologie d'un hôpital de recours à Mascate [Oman], sur une période de cinq ans. Sur un total de 6000 frottis vaginaux, 162 présentaient des résultats histopathologiques concordants [taux de frottis anormaux : 2,7 %] mais 10 d'entre eux n'étaient pas satisfaisants pour le diagnostic histologique. Une corrélation cytohistologique a été observée dans 96/152 des cas [63,2 %], tandis qu'une discordance a été observée pour 56/152 cas [36,8 %]. Des faux négatifs et des faux positifs ont été observés dans 7 cas et 49 cas, respectivement. Les résultats de cette étude confirment le rôle du test de Papanicolaou classique en tant que test de dépistage pour le diagnostic d'une lésion cervicale mais pas pour la prise en charge des patientes. Par rapport à d'autres études, nous avons aussi observé un faible pourcentage de frottis vaginaux anormaux

A specific mitochondrial DNA deletion (mtDNA4977) is identified in a pedigree of a family with hearing loss.

This paper presents a family pedigree of sensorineural hearing loss in patients with a mitochondrial DNA (mtDNA) deletion. Genomic DNA screenings including myo 15 and connexin 26 were normal. MtDNA deletions are associated with many pathophysiologic conditions, including neurological disorders, sensorineural hearing loss, ischemia, cardiomyopathies and aging. Several mitochondrial disorders secondary to mutations or deletions in mtDNA have been identified in association with deafness. The present study describes a pedigree of five individuals with hearing loss who harbor a 4977 bp common aging deletion, in their mtDNA. Chromosomal analysis was normal in all affected individuals. Audiologic and molecular biologic findings of these patients suggest that the common aging deletion of mtDNA may be a predisposing factor in sensorineural hearing loss in this family.

Biologic activity of mitochondrial metabolites on aging and age-related hearing loss.

HYPOTHESIS: Compounds that upregulate mitochondrial function in an aging model will improve hearing and reduce some of the effects of aging. BACKGROUND: Reactive oxygen metabolites (ROM) are known products of oxidative metabolism and are continuously generated in vivo. More than 100 human clinical conditions have been associated with ROM, including atherosclerosis, arthritis, autoimmune diseases, cancers, heart disease, cerebrovascular accidents, and aging. The ROM are extremely reactive and cause extensive DNA, cellular, and tissue damage. Specific deletions within the mitochondrial DNA (mtDNA) occur with increasing frequency in age and presbyacusis. These deletions are the result of chronic exposure to ROM. When enough mtDNA damage accrues, the cell becomes bioenergetically deficient. This mechanism is the basis of the mitochondrial clock theory of aging, also known as the membrane hypothesis of aging. Nutritional compounds have been identified that enhance mitochondrial function and reverse several age-related processes. It is the purpose of this article to describe the effects of two mitochondrial metabolites, alpha-lipoic acid and acetyl L-carnitine, on the preservation of age-related hearing loss. METHODS: Twenty-one Fischer rats, aged 24 months, were divided into three groups: acetyl-l-carnitine, alpha-lipoic acid, and control. The subjects were orally supplemented with either a placebo or one of the two nutritional compounds for 6 weeks. Auditory brainstem response testing was used to obtain baseline and posttreatment hearing thresholds. Cochlear, brain, and skeletal muscle tissues were obtained to assess for mtDNA mutations. RESULTS: The control group demonstrated an expected age-associated threshold deterioration of 3 to 7 dB in the 6-week study. The treated subjects experienced a delay in progression of hearing loss. Acetyl-l-carnitine improved auditory thresholds during the same time period (p<0.05). The mtDNA deletions associated with aging and presbyacusis were reduced in the treated groups in comparison with controls. CONCLUSIONS: These results indicate that in the proposed decline in mitochondrial function with age, senescence may be delayed by treatment with mitochondrial metabolites. Acetyl-l-carnitine and alpha-lipoic acid reduce age-associated deterioration in auditory sensitivity and improve cochlear function. This effect appears to be related to the mitochondrial metabolite ability to protect and repair age-induced cochlear mtDNA damage, thereby upregulating mitochondrial function and improving energy-producing capabilities.

Mitochondrial DNA deletions associated with aging and presbyacusis.

BACKGROUND: The membrane hypothesis of aging proposes an association between reactive oxygen metabolites and aging processes. Reactive oxygen metabolites are a normal by-product of oxidative phosphorylation and are also formed under conditions of ischemia, hypoperfusion, and as a result of environmental contaminants. Among the many detrimental activities of reactive oxygen metabolites, also known as free oxygen radicals, is direct damage to mitochondrial DNA. Progressive accumulation of mitochondrial DNA damage renders cells unable to conduct oxidative phosphorylation reactions effectively, thereby leading to a bioenergetically deficient cell. Over time, mitochondrial DNA damage accumulates and leads to cellular dysfunction with subsequent organ failure, aging, and ultimately, death. This sequence forms the basis of the membrane hypothesis of aging. OBJECTIVE: To determine if the membrane hypothesis of aging may be involved in the development of presbyacusis. DESIGN: Fischer rats from 4 age groups were tested for auditory sensitivity using the auditory brainstem response. Brain, stria vascularis, and auditory nerve tissues were harvested and mitochondrial DNA was amplified to identify the highly conserved cytochrome b and ND1-16S ribosomal RNA segment of the NADH genes, as well as a 4834-base pair (bp) deletion associated with aging. SUBJECTS: Fischer rats (n=28) from 4 age groups were used: young (2-4 months [n=9]), mid-young (9-11 months [n=5]), mid-old (18-20 months [n=5]), and old (30-34 months [n=9]). RESULTS: The results demonstrate a progressive reduction in auditory sensitivity with age. The mitochondrial DNA studies identify a significant increase in the presence of the 4834-bp deletion in the aged subjects compared with the young. CONCLUSIONS: These findings raise the possibility that the 4834-bp deletion may be associated with presbyacusis, as well as with aging.

Mitochondrial DNA deletions associated with aging and possibly presbycusis: a human archival temporal bone study.

HYPOTHESIS: We attempted to determine if the common mitochondrial DNA aging deletion is also associated with presbycusis. BACKGROUND: Presbycusis is the most common cause of deafness in adults in the United States, affecting approximately 40% of the population older than 75 years of age. The ability to identify a gene(s) or a specific genetic deficit(s) associated with presbycusis has significant clinical importance. METHODS: The current study examined mitochondrial DNA (mtDNA) from cochlear sections of 34 human temporal bones: 17 with normal hearing and 17 with presbycusis. DNA was extracted from celloidin-embedded temporal bone sections; and specific oligonucleotide primers were designed to amplify the cytochrome b gene and a 4,977 base pair (bp) deletion of the mtDNA. Polymerase chain reaction (PCR) was used to amplify the base pair products that correspond to targeted gene regions, and sequencing was used to verify the products. RESULTS: Fourteen of the 17 patients with hearing loss showed the 4,977 bp deletion and this deletion was present in only eight of the 17 human specimens with normal audiograms. The cytochrome b gene was amplified from all specimens. CONCLUSIONS: The current study demonstrates the presence of a 4,977 bp deletion in human mitochondrial DNA genome that is associated with aging and with some forms of presbycusis. These results, coupled with previous animal studies, suggest that this 4,977 deletion may be associated with presbycusis.

Association of mitochondrial DNA deletions and cochlear pathology: a molecular biologic tool.

The purpose of these experiments was to develop a method of isolation, amplification, and identification of cochlear mitochondrial DNA (mtDNA) from minute quantities of tissue. Additionally, studies were designed to detect mtDNA deletions (mtDNA del) from the cochlea that previously have been amplified from other organ systems and tissues. MtDNA del have been associated with many pathologies, including neurological disorders, sensorineural hearing loss, ischemia, cardiomyopathies, and aging. DNA was extracted from rat and human tissues, and polymerase chain reaction was used to amplify mtDNA sequences. A 360 base pair (bp) cytochrome-b gene product and the highly conserved ND1-16S ribosomal ribonucleic acid regions found only in mtDNA were amplified from all tissues. Preliminary studies have identified a 4834 bp mtDNA del in aged rats and a corresponding 4977 bp mtDNA del in aged humans. Additionally, preliminary results in human archival temporal bone studies reveal the presence of the 4977-bp mtDNA deletion in two out of three patients with presbycusis. The deletion was not evident in age-matched control patients without a history of presbycusis. This technique of mtDNA identification makes it possible to investigate specific mtDNA defects from a single cochlea, promoting the study of hereditary hearing loss and presbycusis at a molecular biologic level.

SinI modulates the activity of SinR, a developmental switch protein of Bacillus subtilis, by protein-protein interaction.

SinR, a 111-amino-acid DNA-binding protein, is a pleiotropic regulator of several late growth processes in Bacillus subtilis. It acts as a developmental switch, positively regulating genes for competence and motility and repressing aprE and stage II sporulation genes. It is encoded by the second gene in a two gene operon, but previous results have also indicated that these two genes are differently regulated. We show in this discussion that the product of sinI, the first open reading frame (ORF) of this operon, interferes with the function of SinR. In vivo experiments have demonstrated that overexpression of sinI results in phenotypes that are observed in cells with a null mutation of sinR. A chromosomal in-frame deletion of sinI gives rise to a phenotype associated with higher levels of SinR. Thus, SinI acts as an antagonist to SinR. In vitro experiments have shown that the interaction between these two proteins is a direct one. SinI prevents SinR from binding to its target sequence on aprE, and the two proteins form a complex that can be immunoprecipitated with antibodies to either SinR or SinI.

Early spo gene expression in Bacillus subtilis: the role of interrelated signal transduction systems.

The early spo genes are subject to a number of different control mechanisms. We found that at least one histidine kinase, SpoIIJ, is important for the expression of early spo genes but that two others, ComP and DegS, also affect sporulation, especially when SpoIIJ is absent. This indicates the existence of a signal transduction network which may gather information from several sources to feed into the sporulation pathway. Early spo gene expression is inhibited by overproduction of two response regulators, SpoOF and ComA. This effect is eliminated by the elevated presence of their cognate histidine kinases, SpoIIJ and ComP, respectively. This suggests that the unphosphorylated response regulators cause the inhibition of sporulation.

Sin, a stage-specific repressor of cellular differentiation.

Sin is a Bacillus subtilis DNA-binding protein which is essential for competence, motility, and autolysin production but also, if expressed on a multicopy plasmid, is inhibitory to sporulation and alkaline protease synthesis. We have now examined the physiological role of Sin in sporulation and found that this protein specifically represses three stage II sporulation genes (spoIIA, spoIIE, and spoIIG) but not the earlier-acting stage 0 sporulation genes. sin loss-of-function mutations cause higher expression of stage II genes and result in a higher frequency of sporulation, in general. Sin binds to the upstream promoter region of spoIIA in vitro and may thus gate entry into sporulation by directly repressing the transcription of stage II genes. In vivo levels of Sin increase rather than decrease at the time of stage II gene induction, suggesting that posttranslational modification may play a role in downregulation of negative Sin function.

Temporal regulation of the Bacillus subtilis early sporulation gene spo0F.

The initiation of sporulation in Bacillus subtilis depends on seven genes of the spo0 class. One of these, spo0F, codes for a protein of 14,000 daltons. We studied the regulation of spo0F by using spo0F-lacZ translational fusions and also measured Spo0F protein levels by immunoassays. spo0F-lacZ and Spo0F levels increased as the cells entered the stationary phase, and this effect was repressed by glucose and glutamine. Decoyinine, which lowers GTP levels and allows sporulation in the presence of normally repressing levels of glucose, induced spo0F-lacZ expression and raised Spo0F levels. The expression of spo0F-lacZ was dependent on spo0A, -0B, -0E, -0F, and -0H genes, a spo0H deletion causing the strongest effect. In most respects, the spo0F gene was regulated in a manner similar to that of spoVG. However, the presence of an abrB mutation did not relieve the dependence of spo0F gene expression on spo0A, as it does with spoVG (P. Zuber and R. Losick, J. Bacteriol. 169:2223-2230, 1987).

A purified 124-kDa oat phytochrome does not possess a protein kinase activity.

The presence of protein kinase activity in the purified phytochrome preparations [Wong, et al. (1986) J. Biol. Chem. 261, 12089-12097] has been re-examined. The phytochrome preparations having SAR (specific absorbance ratio, A668/A280 for the Pr form as a measure of phytochrome purity) values of greater than 0.95 were homogeneous on SDS gel, but could be further purified to a SAR value of 1.07 by repeated gel filtrations on a Bio-Gel A-0.5 m column. The protein kinase activity remained in the phytochrome preparations having SAR values less than 1.05, but it became undetectable in the phytochrome preparation with a SAR value of 1.07. Two dimensional gel electrophoresis of the phytochrome preparation (SAR, 0.89) showed that a phytochrome band with pl 5.8 had no kinase activity. Phosphorylating activity of the protein kinase was enhanced to some extent by polycations, polylysine and histone. Phytochrome served as a good substrate for this enzyme. The present data indicate that phytochrome has no intrinsic protein kinase activity, but a protein kinase is present in highly purified phytochrome preparations.

An efficient and prolonged in vitro translational system from isolated cucumber etioplasts.

Etioplasts were isolated from dark grown cucumber cotyledons pretreated with kinetin and gibberellic acid. When incubated in a cofactor enriched medium these etioplasts incorporated [35S] methionine into a hot trichloroacetic acid-insoluble fraction; this incorporation was linear for 8 h of incubation and was inhibited by chloramphenicol but not by cycloheximide. Over the same time period, the etioplasts showed continued linear synthesis of the chlorophyll precursors protochlorophyllide, Mg-protoporphyrin and protoporphyrin IX. Analysis of products of in vitro protein synthesis by etioplasts and cotyledons showed the thylakoid membrane polypeptide profiles to be identical. Continued incorporation of [35S] methionine into the large subunit of ribulose bisphosphate carboxylase/oxygenase (RuBisCO) for 8 h has been confirmed further by immunoprecipitation with anti-spinach RuBisCO. This competent in vitro translation system should be useful for future studies of chloroplast protein synthesis and gene expression.

Lochia and menstrual patterns in women with postpartum IUCD insertions.

PIP: 185 postpartum women were inserted with 30-mm Lippes loops on the third or fourth postpartum day to determine the effects of lochia and menstrual patterns. 100 postpartum without IUDs were used as a control group. Hemoglobin and bacteriological studies were done periodically in both groups and amount of lochial discharge and menstrual patterns were recorded. The period of lochia averaged 31.2 days in the study group and 23.3 days in the control group. The amount of lochia was excessive in 40% of the study group as compared with 11% of the controls, and menorrhagia was more often seen in the study group (20% and 7%, respectively). No difference in hemoglobin levels or bacterial count was noted.^ieng

Early post-partum intrauterine contraceptive device insertions.

PIP: 130 Lippes-loop insertions were performed (in India) during early puerperium and patients were followed up for 2-18 months. Of these insertions, 85 were done with the standard Margulies cannula and cervical guard, and in 45 the standard inserter was modified so that the length of the inserter up to the point of ecto-cervix was 5.8 cm, 1 cm longer than the standard length. Patients were aged 15-40 and were para 1-13. Expulsions were 14 and 3 respectively after insertion with the standard and longer instruments. 20 patients who had no living male child accepted the device. Among the 91 patients who returned for follow-up visits 13 complained of menorrhagia. Preliminary results in an associated study of 42 of these women and 42 control women suggest early postpartum IUD insertions are unassociated with infection or postpartum hemorrhage. It is concluded that the postpartum period should be utilized as insertions are easy and painless and expulsions may be minimized by use of a longer inserter.^ieng