A missense mutation encoding cys(67) --> gly in neurophysin ii is associated with early onset autosomal dominant neurohypophyseal diabetes insipidus.

Autosomal dominant neurohypophyseal diabetes insipidus (ADNDI) is an inherited disorder in which progressive degeneration of magnocellular neurons of the hypothalamus impairs production of arginine vasopressin (AVP). ADNDI is caused by mutations in the arginine vasopressin-neurophysin II (AVP-NPII) gene. These mutations are hypothesized to trigger neurodegeneration via disruption of preproAVP-NPII processing. Affected individuals usually develop diabetes insipidus between 1 and 6 years of age. Here we report a novel mutation of the AVP-NPII gene in a family with unusually early presentation of ADNDI. The index case developed symptoms of diabetes insipidus at 1 month of age, her mother at 9 months of age, and the maternal grandfather in early childhood. Each was found to be heterozygous for the missense mutation 1665T > G encoding the amino acid substitution C67G within NPII. This mutation helps to define two homologous regions of the AVP-NPII precursor bounded by disulfide bridges between C13 and C27 and between C61 and C73 that have structural homology and contain the majority of amino acid substitutions associated with ADNDI. The early onset of symptomatic diabetes insipidus in this family suggests that the C67G substitution may be particularly deleterious to magnocellular neurons and may provide a valuable model for study of dominantly inherited neurodegeneration.

Peripartum cardiomyopathy presenting as lower extremity arterial thromboembolism. A case report.

BACKGROUND: Although venous thromboembolism has been associated with peripartum cardiomyopathy, there have been no prior reports of lower extremity arterial thromboembolism complicating cardiac failure. CASE: A 38-year-old woman, gradiva 5, para 5, presented on postpartum day 9 with left pedal parasthesia. Lower extremity angiography found acute thrombotic emboli in the left popliteal artery, right tibial artery and right peroneal artery. When respiratory decompensation ensued, a transthoracic echocardiogram revealed global hypokinesis and a left ventricular ejection fraction of 30%. The patient had an uneventful recovery after treatment with digoxin, furosemide and intravenous heparin. CONCLUSION: Lower extremity arterial thromboembolism may be the initial manifestation of peripartum cardiomyopathy.

Bladder rupture associated with uterine rupture. A report of two cases occurring during vaginal birth after cesarean.

BACKGROUND: Uterine rupture occurs in < 1% of patients undergoing a trial of labor after cesarean section. Associated injury to adjacent organs within the maternal pelvis has likewise been very rarely reported. CASE: Two cases of posterior bladder wall rupture occurred in association with rupture of low transverse uterine incisions. CONCLUSION: Bladder rupture may be associated with uterine rupture during attempted vaginal birth after cesarean. The potential for bladder injury should be included in the patient's antepartum counseling.

In vivo transfer of bacterial marker genes results in differing levels of gene expression and tumor progression in immunocompetent and immunodeficient mice.

To optimize gene delivery for the treatment of malignant mesothelioma, expression of the beta-galactosidase marker gene was examined in a murine model of intraperitoneal malignant mesothelioma. The beta-galactosidase gene was delivered to the peritoneal cavity of tumor-bearing mice by various plasmid-liposome complexes or by replication-incompetent retrovirus, used alone or complexed to liposomes. In tumor samples from immunodeficient nude mice, moderate levels of gene expression were achieved by liposome-complexed plasmids. Retroviral gene delivery was more effective, and was increased nearly 10-fold by complexing the retrovirus to liposomes. In contrast, in tumor samples from immunocompetent CBA mice treated with the same vectors, no marker gene expression was detected. In immunodeficient mice, tumor growth was not affected by beta-galactosidase gene transfer. However, immunocompetent mice showed a significant decrease in tumor size and increase in survival time after beta-galactosidase delivery. Induction of cytotoxic T cells capable of lysing beta-Gal-transfected tumor cells suggests that tumor cells transduced with the bacterial beta-galactosidase gene may be eliminated in immunocompetent hosts. Our findings also indicate that plasmid-liposome complexes, which achieve a low level of gene expression, and retrovirus-liposome complexes, which result in nearly 100 times higher levels of gene expression in tumor cells in vivo, are similarly effective in inducing an antitumor immune response.

The effects of jet nebulisation on cationic liposome-mediated gene transfer in vitro.

Nebulisation is currently the most acceptable and practical delivery system for repeated applications of gene therapy to the lower airways of cystic fibrosis (CF) patients. We have assessed whether this route of administration offers other benefits with regard to respiratory gene transfer. A standard jet nebuliser (Acorn System 22, Medicaid) was used to transfer the reporter gene beta-galactosidase complexed with the cationic liposome DC-Chol/DOPE to three epithelial cell lines in vitro, two non-CF and one CF, using a novel collection system. In all three cell lines, nebulisation resulted in significantly (P < 0.05) improved transfection efficiency compared with instillation. At a constant DNA: liposome ratio of 1:5 (wt:wt), transfection efficiency was inversely related to increasing concentrations of DNA-liposomes before nebulisation. This effect was not related to the amount of DNA delivered and measurements of both zeta potential and mean aerodynamic particle size before and after nebulisation did not show concentration-related differences. The increased transfection efficiency did not relate either to the physical consequences of the nebulisation processes nor the effects of nebulisation on the complexes before instillation. Significantly increased transfection efficiency was seen following nebulisation with 95% O2/5% CO2 in comparison with 21% O2/78% N2 (air); this did not relate to changes in either the pH or temperature of the solution bathing the cells. The data confirm that nebulisation is appropriate for gene delivery to the lower airways in clinical practice and points to factors that may optimise gene transfer efficiency.

Genomic structure and chromosome location of the murine PDE1B phosphodiesterase gene.

Cyclic nucleotide phosphodiesterases (PDEs) catalyze the hydrolysis of cAMP and cGMP, thereby participating in regulation of the intracellular concentrations of these second messengers. The PDE1 family is defined by regulation of activity by calcium and calmodulin. We have cloned and characterized the mouse PDE1B gene, which encodes the 63-kDa calcium/calmodulin-dependent PDE (CaM-PDE), an isozyme that is expressed in the CNS in the olfactory tract, dentate gyrus, and striatum and may participate in learning, memory, and regulation of phosphorylation of DARPP-32 in dopaminergic neurons. We screened an I-129/SvJ mouse genomic library and identified exons 2-13 of the PDE1B gene that span 8.4 kb of genomic DNA. Exons range from 67 to 205 nucleotides and introns from 91 to 2250 nucleotides in length. Exon 1 was not present in the 3 kb of genomic DNA 5' to exon 2 in our clones. The mouse PDE1B gene shares many similar or identical exon boundaries as well as considerable sequence identity with the rat PDE4B and PDE4D genes and the Drosophila dunce cAMP-specific PDE gene dnc, suggesting that these genes all arose from a common ancestor. Using fluorescence in situ hybridization, we localized the PDE1B gene to the distal tip of mouse Chromosome (Chr) 15.

The effect of mucolytic agents on gene transfer across a CF sputum barrier in vitro.

Trials of gene transfer for cystic fibrosis (CF) are currently underway. However, direct application to the airways may be impeded by the presence of airway secretions. We have therefore assessed the effect of CF sputum on the expression of the reporter gene beta-galactosidase complexed with the cationic liposome DC-Chol/DOPE in a number of cell lines in vitro. Transfection was markedly inhibited in the presence of sputum; the effect was concentration dependent and was only partially ameliorated by removal of sputum with phosphate-buffered saline (PBS) washing before gene transfer. However, treatment of the sputum-covered cells with recombinant human DNase (rhDNase, 50 micrograms/ml) but not with N-acetylcysteine, Nacystelyn, lysine (all 20 mM) or recombinant alginase (0.5 U/ml) significantly (P < 0.005) improved gene transfer. Adenovirus-mediated gene transfer efficiency in the presence of sputum was similarly inhibited, and again, treatment with rhDNase before transfection significantly improved gene transfer (P < 0.005). Transfection of Cos 7 cells in the presence of exogenous genomic DNA alone demonstrated similar inhibition to that observed with sputum and was also ameliorated by pre-treatment of DNA-covered cells with rhDNase. In a separate series of experiments performed in the absence of added sputum or genomic DNA, increasing concentrations of rhDNase resulted in a concentration-related decline in transfection efficiency. However, even at the highest concentration (500 micrograms/ml of rhDNase), transfection efficiency remained more than 50% of control. Thus, pre-treatment of CF airways with rhDNase may be appropriate before liposome or adenovirus-mediated gene therapy.

A de novo mutation in the coding sequence for neurophysin-II (Pro24-->Leu) is associated with onset and transmission of autosomal dominant neurohypophyseal diabetes insipidus.

The molecular basis of autosomal dominant neurohypophyseal diabetes insipidus, a hereditary deficiency of vasopressin, was determined by nucleotide sequence analysis of the arginine vasopressin-neurophysin-II gene. A C-->T mutation at nucleotide 1761 was detected in one allele of this gene in each affected individual in three generations of one family. This mutant gene encodes a normal arginine vasopressin peptide, but predicts a substitution of leucine for proline at amino acid 24 of neurophysin-II, the arginine vasopressin carrier protein. This mutation was not detected 50 control individuals, thus demonstrating that it is not a common silent genetic polymorphism. The disease arose in the second generation of the studied family, and the chromosome 20 carrying this new mutation was identified by polymorphic CA microsatellite haplotype analysis. The first affected individual inherited this chromosome segment from her mother, who had neither the disease nor this mutation in her somatic cell DNA. Third generation individuals who subsequently inherited this mutation were affected. These data demonstrate that this amino acid substitution in neurophysin-II causes the disease. Two possibilities to explain the mechanism by which clinical deficiency of arginine vasopressin develops even in the presence of one normal arginine vasopressin-neurophysin-II allele are discussed.

Mechanically induced pelvic pain and organic dysfunction in a patient without low back pain.

Previous reports have identified mechanical disorders of the lumbar spine as a cause of pelvic pain and organic dysfunction (PPOD) in patients with low back pain. Less common however, are reports of mechanically induced pelvic pain and organic dysfunction in patients without accompanying low back pain. This report details the examination findings and treatment response of a patient with pelvic pain, organic dysfunction and clinical evidence of lower sacral nerve root compression (LSNRC) in whom low back pain was not an accompanying finding. Despite the absence of low back pain however, clinical evaluation revealed the characteristic findings of mechanically induced pelvic pain and organic dysfunction secondary to lower sacral nerve root irritation or compression as a result of a mechanical disorder of the low back. As in long standing cases of mechanically induced pelvic pain and organic dysfunction in which low back pain is present, this case also exhibited severe and widespread involvement of the pelvic organs. In spite of numerous failed attempts at treatment directed at the symptomatic component of the patients disorder, complete resolution of symptoms was achieved by manipulative treatment directed at the mechanical disorder of the lumbar spine.

The recognition of mechanically induced pelvic pain and organic dysfunction in the low back pain patient.

Mechanical disorders of the lumbar spine have been identified as a cause of pelvic pain and organic dysfunction (PPOD). Categorically, the clinical features indicative of mechanically induced PPOD fall into three areas: the history of the development or onset of pelvic symptomatology attributable to lower sacral nerve root compression (LSNRC), identification of related symptomatology on presentation, and the recognition of clinical findings indicative of mechanically induced PPOD on examination. Characteristic features of each category are presented. The clinical signs that most reliably indicate the presence of PPOD secondary to a mechanical lesion of the low back are of a sensory nature, and the disappearance or lack of improvement of these signs closely parallels the patient's overall response to manipulative treatment. Without a thorough understanding of the salient features of mechanically induced PPOD, the practitioner is likely to overlook this as a diagnostic possibility. As a result, efforts to document chiropractic spinal manipulative therapy in relieving disorders of pelvic organic function may be hampered. The empirical efficacy of chiropractic spinal manipulative therapy for treating disorders of pelvic organic function would be enhanced if more chiropractors were apprised of the salient features indicating the presence of mechanically induced PPOD.

Chiropractic distractive decompression in treating pelvic pain and multiple system pelvic organic dysfunction.

No data are available on the incidence of pelvic pain and organic dysfunction (PPOD) in patients suffering from low back pain. PPOD is not an uncommon finding in the low back pain patient. Women appear to be more frequently involved than men. The results of recent electrophysiologic investigations indicate that many patients with urological, bowel or anorectal dysfunction demonstrate evidence of denervation neuropathy in muscles innervated by the branches of the pudendal nerve. Six patients with low back pain meeting predetermined criteria, indicating the presence of PPOD as a result of suspected lower sacral nerve root compression (LSNRC) secondary to a mechanical disorder of the low back were treated with chiropractic distractive decompressive manipulation of the lumbar spine. Symptoms of PPOD, and indicators of LSNRC were assessed prior to the onset, and following the termination of treatment. It appears that selected indicators of LSNRC represent the most sensitive clinical signs of identifying the presence of PPOD which may respond to manipulative treatment, and may also provide the most sensitive measure of overall PPOD response. These findings, although preliminary suggest a possible etiology of PPOD in the low back pain patient with evidence of LSNRC. Further work in this area is encouraged.

Chiropractic distractive decompression in the treatment of pelvic pain and organic dysfunction in patients with evidence of lower sacral nerve root compression.

Chiropractic theory postulates that organic dysfunction could be the result of neurological disorganization secondary to mechanical disorders of the spine. Few studies have documented the efficacy of chiropractic manipulative therapy in treating mechanically induced organic dysfunction. Lower sacral nerve root compression (LSNRC) as the result of lumbar disc lesion has been identified as a cause of pelvic pain and organic dysfunction (PPOD). Ten cases of PPOD with accompanying evidence of LSNRC in patients with low back pain as a result of a clinically established lumbar disc lesion are presented with symptomatology prior to and following treatment with distractive decompressive manipulation. A report of one of the cases is detailed. LSNRC is often overlooked as a cause of PPOD. Recognition of associated symptomatology in patients with evidence of LSNRC and confirmation through pain provocation examination is emphasized. Chiropractic distractive decompression may be effective in treating PPOD in patients with evidence of LSNRC as a result of a clinically established lumbar disc lesion.

Pelvic pain and organic dysfunction in a patient with low back pain: response to distractive manipulation: a case presentation.

Many patients with low back pain demonstrate pelvic symptomatology attributable to lower sacral nerve root compression. Lower sacral nerve root compression has been identified as a cause of pelvic pain and pelvic organ dysfunction. Pelvic symptomatology secondary to lower sacral nerve root compression is given. Lower sacral nerve root compression is most commonly the result of lumbosacral disc lesion. A case of low back pain accompanied with pelvic symptomatology is presented along with its response to distractive manipulation. chiropractic treatment may be an effective means of treating pelvic disorders secondary to lower sacral nerve root compression provided that the underlying disc lesion is dealt with, although further study is needed.