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At least 1,711 records · Page 95Linked to original sources
PubMed2003

Parallel down-regulation of chloride channel CLC-K1 and barttin mRNA in the thin ascending limb of the rat nephron by furosemide.

In the past few years the pivotal role of kidney Cl(-)channels (ClC-K) channels in maintaining salt and water homeostasis in the kidney has been established. The aim of the present study was to investigate the influence of the loop diuretic furosemide on the gene expression of the kidney chloride channel ClC-K1 and its recently described functional subunit barttin. Male Sprague Dawley rats received the loop diuretic furosemide (12 mg/kg/day) for 6 days. Rats had free access to 0.9% NaCl, 0.1%KCl solution to prevent volume depletion. Localisation and regulation of ClC-K1 and barttin mRNA was analysed by RNase protection and in situ hybridisation. Nephron-specific regulation was investigated by microdissection and real-time PCR quantification. In furosemide-treated rats ClC-K1 mRNA decreased to half in the inner medulla. In the renal cortex and outer medulla ClC-K1 mRNA levels were weak and did not change. Under furosemide treatment barttin mRNA was regulated in parallel with ClC-K1 mRNA. A significant mRNA decrease occurred after furosemide treatment in inner medulla (0.50 fold), whereas cortical and outer medulla levels remained unaffected. (35)S in situ hybridisation confirmed the regulation and distribution seen in the RNase protection assay experiments. Microdissection of the inner medullary collecting duct and thin limb of Henle's loop followed by real-time PCR revealed that CLC-K1 and barttin mRNA regulation in inner medulla was limited to the thin limb; mRNA levels in collecting ducts were not affected by furosemide treatment. Our findings imply that during furosemide treatment selective down-regulation of ClC-K1 and barttin mRNAs in thin limb plays a role in maintaining salt and water homeostasis.

Actins↗
PubMed1997

Calcium and magnesium transport in the cortical thick ascending limb of Henle's loop: influence of age and gender.

Previous studies from our laboratory have shown that Ca2+ and Mg2+ absorption in the mouse cortical thick ascending limb of Henle's loop (cTAL) is a passive, paracellular process driven by the transepithelial voltage. The passive permeability of the epithelium is enhanced by peptide hormones. The present study investigated whether divalent cation absorption in the cTAL is influenced by cell maturation and/or gender. For this purpose, mouse cTAL segments were microdissected from kidneys of female and male animals aged 4 and 8 weeks. The microdissected tubules were perfused in vitro at a luminal flow rate of 1.5 to 2.5 nl/min. Transepithelial Na+, Cl-, Ca2+ and Mg2+ net fluxes (JX, pmol.min-1.mm-1) were measured using electron microprobe analysis, and the transepithelial potential difference (PDte) was measured continuously. No differences were found in the PDte, JNa and JCl of the various animal groups but the transepithelial Ca2+ and Mg2+ transport capacity of the cTAL was higher in adults (8 weeks) than in young animals (4 weeks). Furthermore, irrespective of age, transepithelial Ca2+ net absorption was greater in male than in female animals. In contrast, the NaCl transport was maximal at 4 weeks in both genders. We conclude therefore that transepithelial divalent cation absorption in the mouse cTAL is an inductive process influenced by cell maturation and gender. The molecular basis of these inductions remains to be elucidated.

Absorption↗
PubMed2001

Influence of dehydration on glycerophosphorylcholine and choline distribution along the rat nephron.

Glycerophosphorylcholine is one of the four major organic osmolytes in renal medullary cells, changing their intracellular osmolyte concentration in parallel with extracellular tonicity during cellular osmoadaptation. In this study, the tubular content of glycerophosphorylcholine was quantified in untreated and 48-h-dehydrated male rats. A chemiluminescence ultra-micromethod was developed to measure choline at the picomolar level in single tubules microdissected from collagenase-treated kidneys. The glycerophosphorylcholine level was calculated as the difference between total choline after acid hydrolysis and the free tubular choline content. In accordance with the glycerophosphorylcholine distribution pattern in different renal zones of untreated rats, low amounts of glycerophosphorylcholine were found in all cortical and outer medullary structures (< 35 pmol/mm), whereas increasing amounts were detected towards the papillary tip (163 pmol/mm). As a percentage of total choline, the level of free tubular choline varied from 4.2% in outer medullary proximal tubules to 30.3% in the inner medullary collecting ducts adjacent to the outer medulla (IMCD1). Antidiuresis led to a nearly twofold increase in glycerophosphorylcholine content in papillary collecting ducts. The osmolality-dependent regulation of organic osmolytes in single microdissected tubules has been demonstrated for the first time. Furthermore, the high tubular glycerophosphorylcholine concentration compared to sorbitol and myo-inositol emphasizes the predominance of glycerophosphorylcholine in the inner medulla and papilla of the rat kidney.

Animals↗
PubMed2006

Analysis of glomerular VEGF mRNA and protein expression in murine mesangioproliferative glomerulonephritis.

Capillary repair is crucial in the healing of glomerulonephritis (GN). The vascular endothelial growth factor (VEGF) has pro-angiogenic properties and plays an important role in glomerular capillary regeneration. Habu Snake Venom (HSV) GN, a murine model for mesangioproliferative GN, was induced in uninephrectomized C57/BL6 mice. Glomerular damage and capillary repair were assessed using morphometry, stereology, and confocal laser scanning microscopy. Mesangiolytic glomeruli were microdissected (days 1,3,7,14) using laser capture microdissection technique. VEGF mRNA expression was analyzed by real-time polymerase chain reaction and compared to intact glomeruli of healthy controls. Spatiotemporal VEGF gene and protein expression was determined using nonradioactive in situ hybridization and immunohistochemistry. On day 1, diseased animals developed focal mesangiolysis paralleled by a significant decrease in length density of glomerular capillaries that gradually returned to baseline levels thereafter, indicating capillary growth in response to initial injury. Glomerular VEGF mRNA expression increased on day 3 and returned back to baseline and beyond at day 14 when the glomerular recovery process was completed. Similarly, glomerular VEGF protein expression tended to be higher on day 3. The present study documents temporarily increased glomerular VEGF gene and protein expression during the healing of HSV GN, suggesting a potential role of VEGF in the repair of mesangiolytic glomerular damage.

Animals↗
PubMed2002

Oligoclonality of a "composite" gastric diffuse large B-cell lymphoma with areas of marginal zone B-cell lymphoma of the mucosa-associated lymphoid tissue type.

We analyzed a case of oligoclonal Helicobacter pylori-associated, primary gastric diffuse large B-cell lymphoma (DLBCL) with areas of extranodal marginal zone B-cell lymphoma (MZBL) of mucosa-associated lymphoid tissue (MALT) type in a 61-year-old male patient. The patient had two separate tumors, in the corpus and in the antrum, each showing two morphologically distinct components (DLBCL and MZBL of MALT type). To determine the clonal relationship between the large- and small-cell components, we microdissected four samples from morphologically distinct tumor components at both tumor sites. PCR analysis revealed rearranged immunoglobulin heavy chain (IgH) genes of different sizes in three of the four microdissected samples. Cloning and sequencing of the PCR products displayed different IgH gene rearrangements in the two small cell components of the antrum and the corpus. A third genuinely differently rearranged IgH gene was found in the large-cell components of both antrum and corpus. Our results suggest that in primary gastric DLBCL with areas of MZBL of MALT type, the small-cell component may comprise more than one tumor clone. Furthermore, the large-cell component may evolve independently from coexisting MZBL.

Amino Acid Sequence↗
PubMed2002

Human hormone-refractory prostate cancers can harbor mutations in the O(2)-dependent degradation domain of hypoxia inducible factor-1alpha (HIF-1alpha).

PURPOSE: Androgen ablation, the preferred therapy for advanced prostate cancer, reduces blood flow and induces hypoxia in androgen-dependent tissues. Given the transient effectiveness of this therapy, we must consider whether a hypoxia-resistance mechanism might be involved in the development of therapeutic resistance by prostate cancer cells. The transcription factor protein, hypoxia-inducible factor 1alpha (HIF-1alpha), helps increase the expression of gene products that enable cells to survive conditions of hypoxic stress. Enhanced HIF-1alpha expression during hypoxia results from a drastic reduction of its degradation rate within a critical region of the protein referred to as the "oxygen-dependent degradation (ODD) domain". We sequenced HIF-1alpha cDNAs amplified from human prostate cancer cell lines and from hormone resistant prostate cancer specimens to determine whether prostate cancer cells might harbor mutations within the HIF-1alpha ODD domain. METHODS: HIF-1alpha cDNAs were RT-PCR amplified from three prostate cancer cell lines (LNCaP, PC-3, and DU145), from five different human hormone-resistant prostate cancers and one normal prostate, all microdissected, and were sequenced to determine whether the HIF-1alpha gene products were wildtype or mutant. One specimen containing a hormone-resistant prostate tumor that expressed a mutated HIF-1alpha cDNA was further microdissected into benign and tumorous regions and DNAs extracted from these regions were directly amplified by PCR and sequenced to determine whether the HIF-1alpha mutation was specific to the tumor. RESULTS: Although the HIF-1alpha cDNAs of all cell lines, the normal prostate, and three of the tumors were found to have a wildtype sequence, HIF-1alpha cDNAs amplified from two hormone-resistant tumors had nucleic acid substitutions that resulted in significant amino acid changes within the ODD domain of the HIF-1alpha protein. Analysis of the DNA extracted from a benign or tumorous region of one of these specimens showed that only the wildtype (nonmutated) form of the HIF-1alpha gene was amplified from the normal DNA whereas only the mutated form of the HIF-1alpha gene was amplified from the tumor. CONCLUSIONS: Some human hormone-refractory prostate cancers have mutations in a critical regulatory domain of the HIF-1alpha protein. We believe that these mutations might enable expression of this protein under inappropriate conditions and contribute to the development of therapeutic resistance by the cancer cells. This hypothesis is currently being tested.

Base Sequence↗
PubMed2005

Comparative genomic hybridization analysis for pancreatic cancer specimens obtained by endoscopic ultrasonography-guided fine-needle aspiration.

BACKGROUND: Comparative genomic hybridization (CGH) analysis of pancreatic cancer has been done exclusively for surgical and autopsy specimens, because of the difficulty of tissue sampling without surgery. To overcome this difficulty, we applied CGH technology to cells obtained by endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA). METHODS: In the present study, we performed EUS-FNA for 17 patients with pancreatic cancer before surgery. Tumor cells were selected by microdissection. DNA was extracted from the cells and amplified by degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR). Then CGH was carried out. RESULTS: In the 15 patients with tubular adenocarcinoma, the most common loci of gains (including amplification) were 5p, 8q, and 20q (60% of the patients); and 1q, 7p, and 12p (27%). The most frequent losses were 17p (73%); 9p, 18q, and 19p (47%); and 8p (33%). These findings were similar to our previously reported data. Both of the patients with acinar cell carcinoma showed gains of 2q and 5p, and losses of 1p, 9p, 9q, 11p, 11q, 14q, 17p, 17q, and 18q. CONCLUSIONS: The results of this study suggest that comprehensive genetic analysis is possible for EUS-FNA biopsy specimens, with a combination of microdissection and DOP-PCR. This analytical strategy will enable us to evaluate the biological characteristics of pancreatic cancer before treatment.

Adenocarcinoma↗
PubMed2005

Expression profiling using human tissues in combination with RNA amplification and microarray analysis: assessment of Langerhans cell histiocytosis.

Advances in molecular genetics have led to sequencing of the human genome, and expression data is becoming available for many diverse tissues throughout the body, allowing for exciting hypothesis testing of critical concepts such as development, differentiation, homeostasis, and ultimately, disease pathogenesis. At present, an optimal methodology to assess gene expression is to evaluate single cells, either identified physiologically in living preparations, or by immunocytochemical or histochemical procedures in fixed cells in vitro or in vivo. Unfortunately, the quantity of RNA harvested from a single cell is not sufficient for standard RNA extraction methods. Therefore, exponential polymerase-chain reaction (PCR) based analyses, and linear RNA amplification including amplified antisense (aRNA) RNA amplification and a newly developed terminal continuation (TC) RNA amplification methodology have been used in combination with microdissection procedures such as laser capture microdissection (LCM) to enable the use of microarray platforms within individual populations of cells obtained from a variety of human tissue sources such as biopsy-derived samples {including Langerhans cell histiocytosis (LCH)} as well as postmortem brain samples for high throughput expression profiling and related downstream genetic analyses.

Gene Expression Profiling↗
PubMed1999

Atrophic gastritis, Epstein-Barr virus infection, and Epstein-Barr virus-associated gastric carcinoma.

BACKGROUND: The developmental process of Epstein-Barr virus (EBV)-associated gastric carcinoma (EBVaGC) has not been clearly demonstrated, especially in its relation to intestinal metaplasia and epithelial EBV infection.METHODS: Gastritis and intestinal metaplasia was histologically evaluated in non-neoplastic gastric mucosa that surrounded early carcinoma of EBVaGCs ( n = 23) and EBV-negative gastric carcinomas (GCs) (intestinal type, n = 139; diffuse type, n = 44). Helicobacter pylori infection was evaluated by immunohistochemistry. EBV infection in the gastric mucosa was examined by both RNA probe in situ hybridization (ISH) and polymerase chain reaction (PCR) for the BamHI-W region of EBV DNA, the latter of which was applied to the microdissected mucosa.RESULTS: Marked grade of atrophy and moderate to marked grade of lymphocytic infiltration were significantly more frequent in EBVaGCs (74% and 78%, respectively), compared to intestinal-type (49% and 12%)and diffuse-type (27% and 12%) of EBV-negative GCs. Only 13% of EBVaGCs were surrounded by intestinal metaplasia, in contrast to 41% of intestinal-type EBV-negative GCs. Immunohistochemistry revealed nearly the same frequencies of H. pylori infection (70%) in three types of GCs. RNA probe ISH for EBV-DNA failed to identify any positive cells in nonneoplastic mucosa, including intestinal metaplasia. Two of 118 microdissected samples of EBVaGC and 5 of 62 samples of EBV-negative GCs showed amplification of EBV-DNA, consisting of 3 pyloric and 4 fundic but no metaplastic gland samples.CONCLUSIONS: EBVaGC may develop from rare EBV-infected epithelial cells with severe atrophic gastritis, but the process is not directly related to intestinal metaplasia or H. pylori infection.

Journal Article↗
PubMed2005

Papillary lesions of the breast: a molecular progression?

INTRODUCTION: Breast papillary lesions represent a heterogeneous group of tumors ranging from benign to malignant, including several intermediate forms. Malignant papillary tumors are rare and their molecular characterization is still limited. A few studies pointed to the presence of specific genetic alterations that could be relevant both for diagnostic purposes and to elucidate tumour development and progression. In order to look into the issue, we compared LOH relative frequencies of four microsatellite markers located on chromosome 16 in a set of morphologically different papillary breast lesions. LOH at TP53 locus was also analyzed throughout lesions. MATERIALS AND METHODS: Fifteen cases were analyzed. Sections including a malignant papillary lesion, a benign lesion (when available), and normal breast tissue were selected. Fifteen malignant and twelve benign areas were microdissected using the Leica laser microdissection system (AS LMD). After DNA extraction samples were tested for the following markers: TP53, D16S423, D16S310, DS163210 and D16S476, and analyzed on ABI PRISM 3100 (Applied Biosystems, Foster city CA). RESULTS: Fourteen malignant lesions and twelve paired benign areas appeared to be informative for at least one of the four markers on chromosome 16. In particular, LOH at loci 16p13 and 16q21 was detected in both benign and malignant lesions, whereas LOH at locus 16q23 was limited to malignant lesions. Nine malignant and seven benign lesions were informative for LOH at TP53 locus, that was found to be significantly associated (p=0.01) with the malignant phenotype. CONCLUSIONS: Our data suggest an involvement of chromosome 16 mutations in the early steps of breast papillary tumorigenesis. TP53 deletion and possibly LOH at 16q23 appear to play a role as progression factors, being they significantly associated with malignant transformation of breast papilloma.

Biomarkers, Tumor↗
PubMed2005

Unusual distribution pattern of telomeric repeats in the shrews Sorex araneus and Sorex granarius.

Sorex araneus and Sorex granarius are sibling species within the Sorex araneus group with karyotypes composed of almost identical chromosome arms. S. granarius has a largely acrocentric karyotype, while, in S. araneus, various of these acrocentrics have combined together by Robertsonian (Rb) fusions to form metacentrics, with the numbers and types of metacentrics differing between chromosomal races. Our studies on telomeric sequences in S. araneus and S. granarius revealed differences between chromosomes and between species. In S. araneus (the Novosibirsk race), hybridization signals were present on the telomeres of all the chromosomes after FISH with a PCR-generated telomeric probe. In addition, hybridization signals were observed at high frequencies in the pericentric regions of some but not all metacentrics formed by Rb fusion. There were fewer signals on those metacentrics formed earlier in the evolution of S. araneus. This suggests that S. araneus chromosomes retain at least some telomeric repeats during Rb fusion, but that these repeats are lost or modified over time. These results are critical for the interpretation of the well-studied hybrid zones between chromosomal races of S. araneus, given that Rb fission has been postulated in such hybrid zones and that the likelihood of Rb fission will relate to presence/absence of telomeric sequences at the centromeres of metacentrics. In S. granarius, there were strong signals at the proximal (centromeric) telomeres of the acrocentrics after FISH with a DNA telomeric probe. FISH with a PNA telomeric probe on S. granarius acrocentrics showed that the proximal telomeres were 213 kb on average, while the length of the distal telomeres was 3.8 kb on average. Two-colour FISH, using a telomeric DNA probe and a microdissected probe generated from the pericentric regions of the S. granarius chromosomes a and b, revealed regions on distinct chromatin fibres where telomeric and microdissected probes were colocalized or localized sequentially. The proximal telomeres of S. granarius are highly unusual both in their large size and their heterogeneous structure relative to the telomeres of other mammals.

Animals↗
PubMed1983

Presence of low molecular weight polypeptides in human brunescent cataracts.

Microdissected sections from opaque & brunescent lens nuclei contain low molecular weight (4,000-8,000 dalton) polypeptides not found in microdissected sections of transparent lens nuclei. Tryptic digestion of these polypeptides from different cataracts reveal similar peptide maps. Together, these results support the involvement of proteolysis in human cataractogenesis, and suggest the possibility of similar molecular mechanisms occurring in cleavage of lens polypeptides during formation of the opaque & brunescent human cataract.

Aged↗
PubMed1985

Distribution of dopamine, noradrenaline and adrenaline in coronal sections of the rat lower brainstem.

The concentration of the three major catecholamines (CAs) were determined in 500 micron thick coronal sections of the rat medulla oblongata dissected into microcubes. Noradrenaline (NA) concentrations were always found much higher than the levels of the two other CAs in the same microcube. The highest concentrations of the three CAs were found in the dorso-medial region of the lower brainstem, more exactly in the more caudally located parts of the nucleus tractus solitarii (NTS). In the ventro-lateral region, the CA concentrations were lower and, except for adrenaline (A), did not exhibit any substantial change in their rostro-caudal distribution. Conversely, in the dorso-medial region, there was a clear rostro-caudal pattern of distribution of the three CAs. This distribution was similar for the three amines, since only a small difference (about 500 micron) was found between the maximal NA and A concentrations. Since the three CAs are present in highest concentrations within the same dorso-medial or ventro-lateral groups of microcubes, a microdissection of these two areas seems suitable to study simultaneously the metabolism of the three CAs in the rat lower brainstem. These data also suggest a microdissection procedure to study A metabolism within the C2-C3 A cell bodies and within a region more caudally located, rich in A terminals.

Adrenergic Fibers↗
PubMed1996

GABAergic neuronal activity and mRNA levels for both forms of glutamic acid decarboxylase (GAD65 and GAD67) are reduced in the diagonal band of Broca during the afternoon of proestrus.

There is considerable evidence that GABAergic neurons play an important role in the regulation of gonadotropin-releasing hormone (GnRH) secretion, and that these neurons may mediate the feedback actions of gonadal steroids on GnRH neurons. The aim of the present study was to investigate whether endogenous changes in ovarian steroid secretion during the estrous cycle influenced GABAergic neuronal activity in the preoptic region of the hypothalamus, and in other steroid-sensitive brain regions. Intact, adult female rats were sacrificed at various times during the days of metestrus or proestrus. GABAergic neuronal activity was estimated by measuring the rate of accumulation of GABA in microdissected brain regions after pharmacological inhibition of GABA degradation. Concentrations of mRNA for both forms of glutamic acid decarboxylase (GAD65 and GAD67) were quantified in microdissected brain regions by a microlysate ribonuclease protection assay. In the diagonal band of Broca at the level of the organum vasculosum of the lamina terminalis (DBB(ovlt)), GABAergic neuronal activity was significantly reduced during the afternoon of proestrus compared with the morning of either proestrus or metestrus. In the lateral septal nucleus, GABAergic neuronal activity was significantly increased in the afternoon of proestrus compared with the morning. There were no significant effects of time of day or day of estrous cycle in the medial preoptic nucleus, median eminence, ventromedial nucleus, suprachiasmatic nucleus, medial septal nucleus, hippocampus (CA1 region), or cingulate cortex. In the DBB(ovlt), mRNA levels for both GAD65 and GAD67 were significantly reduced in the afternoon of proestrus compared with the afternoon of metestrus. By contrast, there was no change in GAD65 and GAD67 mRNA levels in the cingulate cortex at any of the times examined. These results demonstrate that GABAergic neuronal activity, and mRNA levels for both GAD65 and GAD67, are reduced in the DBB(ovlt) during the afternoon of proestrus. These results support the hypothesis that decreased GABAergic neuronal activity in this region plays a major permissive role in the generation and maintenance of the estrogen-induced LH surge.

Animals↗
PubMed1991

Morphometry and cell proliferation in endoscopic biopsies: evaluation of a technique.

A simple method for the quantification of intestinal epithelial cell proliferation in humans was evaluated. Endoscopic biopsy specimens were stained and microdissected, and the number of mitoses per gland or crypt was determined, as was the area of these units. The technique could readily be applied to tissue from the stomach, small intestine colon, and rectum. The number of mitoses and the size of the proliferative compartments increased caudally from the stomach to the cecum in humans. There was a good correlation between area and mitoses per gland or per crypt (r = 0.89; P less than 0.001), confirming the general equivalence between proliferative rate and compartment size. The method was validated by comparing microdissection-derived data with data previously obtained as part of an autoradiography-based study in the dog. This showed that similar differences in proliferation and crypt cell mass could be obtained but in less than one sixth of the time taken to score autoradiographs. It is concluded that this method for the estimation of gastrointestinal epithelial proliferation correlates well with other well-established techniques, confers speed as well as accuracy, has an all encompassing denominator, and can thus avoid many of the problems associated with the quantification of tissue sections.

Animals↗
PubMed1988

Retrotransposon-like sequences are expressed in Y chromosomal lampbrush loops of Drosophila hydei.

The DNA sequence family micropia consists of repeated DNA sequences that occur dispersed in the genome of Drosophila hydei. Members of this DNA sequence family were recovered from two recombinant DNA clone banks obtained by microdissection of the two Y chromosomal lampbrush loop threads and pseudonucleolus from primary spermatocyte nuclei. Nucleotide sequence analysis of two of the recombinant DNA clones revealed homology to the DNA region coding for a reverse transcriptase-like protein in retroviruses and retrotransposons. Homologous tissue-specific transcripts of a size of 1.2 x 10(3) base-pairs were found in testes. Transcript in-situ hybridization shows that at least parts of these transcripts are synthesized in these Y chromosomal lampbrush loops, which were originally used for microdissection. Also the cytoplasm of primary spermatocytes contains homologous RNA species. These observations are discussed in the context of lampbrush loop function and evolution.

Amino Acid Sequence↗
PubMed1990

Unique alterations of neuropeptide content in median eminence, amygdala, and dorsal vagal complex of 3- and 6-week-old diabetes mutant mice.

The nature of the genetic defects which define the obese (ob) and diabetes (db) loci in mice remain unknown, but both produce similar syndromes when maintained in the same strain of mice. There is some evidence suggesting a lesion in the central nervous system (CNS) in db/db mice, while ob/ob mice appear to have a primary lesion outside the CNS. In a search for further evidence of a unique central lesion in db/db mice, we have examined neuropeptide content in selected, microdissected brain areas in both of these mutants and lean controls. In order to rule out possible interactions of the db mutation with the genetic background, diabetes mice of both C57BL/KsJ and C57BL/6J strains were studied. When concentrations of nine neuropeptide immunoreactivities were examined in up to seven microdissected areas of the brain, C57BL/6J ob/ob mice showed only one reproducible alteration, a lower content of beta-endorphin-like immunoreactivity (LI) in the preoptic area at both 3 and 6 weeks of age as compared with lean littermates. In contrast, db/db mice of both C57BL/6J and C57BL/KsJ strains exhibited alterations in a total of four peptides in three brain areas: lower concentration of somatostatin-LI in median eminence, higher Met-enkephalin-LI in dorsal vagal complex of the medulla oblongata, higher substance P-LI and lower vasoactive intestinal polypeptide (VIP)-LI in amygdala. The concentrations of the peptides studied in medial basal hypothalamus, lateral hypothalamus, substantia nigra, and preoptic area were not reproducibly altered in db/db mice. These data provide preliminary evidence for unique brain abnormalities in db/db mice in specific areas that are involved in processing of neural signals that can affect the islets of Langerhans, gonadotrophin secretory patterns, and many other visceral functions.(ABSTRACT TRUNCATED AT 250 WORDS)

Amygdala↗
PubMed1996

Absence of papillomavirus DNA in normal tissue adjacent to most cervical intraepithelial neoplasms.

OBJECTIVE: To determine if human papillomavirus (HPV) DNA is present in the normal mucosa adjacent to cervical intraepithelial neoplasia (CIN). METHODS: Serial sections of 28 CIN lesions were studied. Lesional and normal epithelia and stroma were microdissected; the DNA was extracted and amplified by polymerase chain reaction (PCR) using primers designed to amplify both HPV late (L1) and human beta-globin sequences. Human papillomavirus was typed by restriction fragment length polymorphism analysis following digestion of PCR products. RESULTS: Twenty-five of 28 (89%) lesional epithelia scored positive for HPV nucleic acids. In' four of 25 (16%) HPV-positives, the normal squamous epithelium scored positive for HPV nucleic acids, two of which (8%) also scored positive in the stroma. Repeat microdissection and PCR analysis of three of these cases was performed and all were negative in both normal epithelium and stroma, suggesting laboratory contamination. CONCLUSION: Human papillomavirus nucleic acids are present uncommonly in normal-appearing squamous epithelium adjacent to CIN. This does not exclude occult infection in the natural history of CIN but indicates that when lesions develop, occult infection is not normally maintained in the normal mucosa. This is consistent with the low recurrence rates following ablation as well as low indices of HPV positivity in normal cervices during follow-up. This finding should be taken into consideration when counseling patients and is relevant to the concept of HPV testing during follow-up after cone biopsy.

DNA Probes, HPV↗