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Diagnosis of type A Niemann-Pick's disease by conjunctival biopsy.

Electron microscopic examination of the conjunctive of four children affected with Niemann-Pick's disease revealed a widespread storage process. The epithelium, the connective tissue cells, the endothelial cells and the pericytes of the capillaries are filled with lysosomal inclusions with pleomorphic structure. The nerve fibres show destructive process of the myelin sheaths and accumulation of lipid material in the Schwann cells. The extension of the lesions is similar, as shown by morphologic and morphometric analysis in early and terminal stages of the diseases. Conjunctival biopsy allows an early and secure diagnosis of Niemann-Pick's disease, and should be performed when the clinical symptoms suggest a lysosomal disorder.

Biopsy↗

Avian pox in blue-fronted Amazon parrots.

During a 1-month period at a quarantine station, an epornitic of avian pox occurred in blue-fronted Amazon parrots (Amazona aestiva). Clinical signs included conjunctivitis, blepharitis, and varying degrees of anorexia and respiratory distress. Lesions included periocular ulcerations and scabs and necrotic plaques in the oral cavity. Histologically, the lesions consisted of epithelial hyperplasia, secondary inflammatory changes, and eosinophilic inclusions which, by electron microscopy, were shown to contain poxvirus. When chicken embryos were inoculated with material from eyelid scabs and pharyngeal plaques, lesions of avian pox developed on the chorioallantoic membrane. The death rate of infected birds was high because of secondary bacterial and fungal infections, but uncomplicated cases were usually self-limiting. Periocular lesions also developed in 2 other species of psittacine birds housed in the same facility.

Animals↗

Arthroscopic distal clavicle resection from a bursal approach.

We retrospectively reviewed 117 consecutive patients who underwent arthroscopic acromioclavicular joint (ACJ) arthroplasties. Only patients who underwent ACJ arthroplasties from a bursal approach in conjunction with subacromial decompression were included. Patients with isolated ACJ arthrosis treated with resection of the distal clavicle from a superior approach, isolated impingement with only undersurface distal clavicle debridement, prior surgery, or other shoulder pathology were excluded. Twenty-four patients met these rigid criteria for inclusion in the study. After an arthroscopic subacromial decompression, the distal clavicle was visualized and resected through a standard bursal approach. In addition, an anterosuperior portal was used in 50% of the patients to confirm adequate clavicle resection. Postoperative follow-up averaged 32.5 months (range, 24 to 70 months). Preoperative and postoperative pain were rated subjectively on a 5-point scale (1, incapacitating pain; 5, no pain). Operative reports and postoperative radiographs were reviewed to determine technical factors that may have influenced outcome. Seventeen patients had excellent results (71%), 4 good (16.5%), and there were 3 failures (12.5%). Average preoperative pain rating was 1.8 and was improved to 4.3 postoperatively. The average amount of clavicle resection was only 5.4 mm. Given smooth, even, and complete bone removal, the amount of bone resected did not correlate with outcome. Arthroscopic distal clavicle resection performed in conjunction with subacromial decompression gave excellent results, comparable to isolated ACJ procedures. In this series, additional use of an anterosuperior portal for more direct shaver placement and complete ACJ viewing allowed consistent bone resection and excellent results in a high percentage of patients.

Acromioclavicular Joint↗

Selectivity in capillary electrochromatography using native and single isomer anionic cyclodextrin reagents.

Separations of naphthalene compounds that differ in position of substitution and type of substituent were accomplished using cyclodextrin distribution capillary electrochromatography. Separation systems composed of running buffers containing mixtures of native neutral and single isomer anionic cyclodextrins (CDs) were employed yielding efficiencies of approximately 200,000 plates/meter. Solute migration rates and relative orders can be readily modified by changing CD types and concentrations. Experiments were performed to determine distribution coefficients between each of the CDs used in these studies and an aqueous running buffer. For this work, naphthalene-CD cavity inclusion is assumed to be the principal mode of interaction. The distribution coefficients for carboxymethyl-beta-cyclodextrin (CM-beta-CD), degree of substitution 1, were 10-70% larger than those for native beta-CD and 75-1800% large than those for gamma-CD. The CM-beta-CD was singly charged and yielded a narrow elution window. Nevertheless, baseline resolution was achieved for several substituted naphthalene compounds using CM-beta-CD in conjunction with beta-CD or gamma-CD. Under certain conditions, the gamma-CD system yielded an elution order that differed from that of the beta-CD system. Heptakis-(2,3-dimethyl-6-sulfato)-beta-CD with its -7 charge produced a much larger elution window. The extensive substitution with sulfonic groups at the truncated bottom of the CD seemed to inhibit inclusion as the distribution coefficients for the naphthalene compounds were generally more than an order of magnitude smaller than those for CM-beta-CD. Moreover, there was evidence that this sulfato-CD interacted with both the capillary wall and neutral beta-CD. This work differs from prior uses of CDs in that relatively complicated mixtures of neutral, achiral compounds are separated using combinations of recently developed single-isomer CDs as running-buffer additives. The single-isomer CDs, as opposed to most highly complex derivatized CD products, facilitate predictions of separation performance for multicomponent samples. In this manner, the ability to use knowledge of distribution coefficients to predict elution characteristics for a ternary CD system is demonstrated.

Anions↗

Conjunctival scrapings and impression cytology in chronic conjunctivitis. Correlation with microbiology.

To assess the utility of exfoliative cytology in the diagnosis of chronic conjunctivitis, we compared its findings with microbiological examination and established the correlation between cytology and microbiology in 32 patients with conjunctivitis lasting four weeks or more, with a nonspecific clinical picture. The material for cytology was collected by scraping and by imprint of the ocular surface on Millipore filter paper. The specimens were stained with Giemsa. Material for bacterial culture and fluorescent antibody test for viruses and Chlamydia were obtained at the same time. In 12 patients neutrophils predominated in cytology, in six eosinophils were found and in four there were more lymphocytes. One patient had a basophilic intracytoplasmic inclusion specific for Chlamydia infection. In eight patients coagulase-negative Staphylococcus species and in one Staphylococcus aureus were isolated, in two the fluorescent antibody test for Chlamydia was positive and in three for viruses. In all patients with a positive bacterial culture, a predominance of neutrophils was observed. All patients with a positive test for viruses had lymphocytosis in conjunctival scrapings. In two patients microbiology detected Chlamydia, but characteristic intracytoplasmic inclusions were observed only in one. Scrapings gave more specific results, that correlated well with microbiology in bacterial and viral infection, whereas the results of impression cytology showed minor metaplastic changes of epithelial cells that were nonspecific for the type of inflammation. Cytology remains a useful non-invasive tool and is an adjunct to clinical and microbiological examination. It proved especially helpful in allergic conjunctivitis.

Antibodies, Bacterial↗

Comparison of pre-S1 and pre-S2 proteins in hepatocytes with replication by in situ hybridization assay.

To compare the relationship of pre-S1 and pre-S2 proteins in hepatocytes with HBV replication, HBVDNA in situ hybridization was performed for a group of patients with chronic hepatitis by bio-probe, in conjunction with detection of intrahepatic HBsAg, HBcAg, pre-S1 and-S2 antigens. It was found that the expression of intrahepatic pre-s1 and pre-S2 proteins was visualized as cytoplasmic homogeneous and inclusion types, on some occasions as membranous localization. However, the positive rate for pre-S1 protein was up to 75% (15/20), being significantly higher than that of pre-S2 protein (35%; 7/20) in the same group (P less than 0.05). What in more important, the pre-S1 expression, rather than pre-s2, was in parallel with the presence of intrahepatic HBVDNA and HbcAg. All these findings suggest that the pre-S1 protein might be mainly expressed at the HBV replication phase, thus, to some extent, the detection of pre-S1 protein might complement HBe system and HBVDNA clinical routine.

Biomarkers↗

The indications for lumbar and cervical disc replacement.

Although cervical and lumbar disc replacements are in their spinal surgical infancy with regard to clinical application, the indications are already delineated. Lumbar disc arthroplasty is indicated for one- or two-level discogenic mechanical back pain primarily in the absence of radiculopathy. In contrast, cervical disc replacement can be readily applied in patients presenting with neurologic deficit, radiculopathy or myelopathy because the approach and anterior spinal cord decompression are identical for anterior cervical disc replacement and traditional Smith-Robinson cervical decompression. In addition, the application of more complex spinal osteotomies, revision of pseudarthroses and deformity correction are much more applicable to the cervical arthroplasty procedures. This is because even the most experienced vascular access surgeon has difficulty with the formidable revision through a repeat anterior lumbar procedure, whereas most experienced cervical spinal surgeons are familiar with repeat anterior cervical approaches. The end result is that surgeons will have more trepidation with multilevel lumbar arthroplasties, especially those presenting in conjunction with neurologic symptoms. At the current time radiculopathy is an exclusion criteria for the four prospective Investigational Device Exemption (IDE) Food and Drug Administration (FDA) studies on lumbar disc replacement, whereas cervical radiculopathy is an inclusion criteria for the major IDE FDA investigations of cervical arthroplasties.

Arthroplasty, Replacement↗

NMR and modelling studies of disaccharide conformation.

Long-range heteronuclear coupling constants were measured across the glycosidic linkages for a series of eight alpha- or beta-linked disaccharides in aqueous solution. Multiple 13C site-selective excitation experiments using 1H decoupling in conjunction with pulsed field gradient-enhanced spectroscopy were used to determine 3J(C,H) values. These were subsequently compared with the respective couplings calculated, using a Karplus relationship, from molecular dynamics simulations with the explicit inclusion of water.

Carbohydrate Conformation↗

Interaction of muscleblind, CUG-BP1 and hnRNP H proteins in DM1-associated aberrant IR splicing.

In myotonic dystrophy (DM1), both inactivation of muscleblind proteins and increased levels of CUG-BP1 are reported. These events have been shown to contribute independently to aberrant splicing of a subset RNAs. We demonstrate that steady-state levels of the splice regulator, hnRNP H, are elevated in DM1 myoblasts and that increased hnRNP H levels in normal myoblasts results in the inhibition of insulin receptor (IR) exon 11 splicing in a manner similar to that observed in DM1. In normal myoblasts, overexpression of either hnRNP H or CUG-BP1 results in the formation of an RNA-dependent suppressor complex consisting of both hnRNP H and CUG-BP1, which is required to maximally inhibit IR exon 11 inclusion. Elevated levels of MBNL1 show RNA-independent interaction with hnRNP H and dampen the inhibitory activity of increased hnRNP H levels on IR splicing in normal myoblasts. In DM1 myoblasts, overexpression of MBNL1 in conjunction with si-RNA mediated depletion of hnRNP H contributes to partial rescue of the IR splicing defect. These data demonstrate that coordinated physical and functional interactions between hnRNP H, CUG-BP1 and MBNL1 dictate IR splicing in normal and DM1 myoblasts.

Base Sequence↗

[Ultrastructural manifestations of early metabolic disorders in the myocardium of dogs with alloxan diabetes].

The ultrastructure of dog cardiomyocytes was studied one month after reproducing alloxan diabetes, with respect to myocardial extraction from the blood of glucose, nonesterified fatty acids, beta-lipoproteins and ketone bodies as well as to respiration in conjunction with oxidative phosphorylation of mitochondria, hexokinase and phosphorylase activity. Destructive changes in mitochondria, increased glycogen content in cardiomyocytes were revealed in the absence of glucose consumption by the myocardium which absorbed only lipoid metabolites. Lipoid inclusions were rarely seen in cardiomyocytes. The myocardium showed the increased content of lysosomes and hydrolytic phosphorylase. It is suggested that lipoid metabolites transform to glycogen, with lysosomes participating in the process.

Alloxan↗

Use of endometrial measurement as an exclusion criterion for in vitro fertilization using clomiphene citrate.

OBJECTIVE: Previous reports have indicated an association between endometrial development and pregnancy outcome for patients treated with clomiphene citrate (CC) in conjunction with intrauterine insemination or intercourse. We expanded the use of CC for ovulation induction in association with in vitro fertilization (IVF). This study was designed to determine if endometrial thickness should be used as an inclusion or exclusion criterion for CC-IVF. STUDY DESIGN: One hundred twenty-eight patients were enrolled in an ovulation-induction regimen using CC for expected IVF-ET between January 1992 and December 1992. A total of 81 patients met inclusion criteria for CC-IVF and had endometrial measurement performed prior to human chorionic gonadotropin administration. Patients were categorized on the basis of endometrial measurement as follows: (A) > 4 - < 7 mm, (B) > or = 7 - < or = 10 mm, and (C) > 10 mm. Standard IVF was performed, and pregnancy rates for each category were evaluated. RESULTS: A total of 23 pregnancies (28% per retrieval) were established. Pregnancy rates were not different by category (P > .10, Fisher's Exact Test): (A) 3/15 (20%), (B) 13/41 (32%), and (C) 7/25 (28%). CONCLUSION: These data suggest that for CC-IVF. endometrial measurement should not be used as an exclusion criterion since pregnancies occurred at comparable frequencies in all the groups.

Adult↗

Foreign-body inclusion cyst presenting on the lateral nasal sidewall 1 year after rhinoplasty.

Hemostasis and decreased risk of synechiae formation are benefits of nasal packing after sinus surgery; however, these must be weighed against the possible complications, which can include devastating toxic shock syndrome. Nasal packing is often impregnated with an ointment that allows for a less traumatic placement into and removal from the nasal cavity, as well as providing some antibiotic coverage for nasal bacterial flora. Orbital complications secondary to the petroleum-based packing have been reported. When sinus surgery is performed in conjunction with a rhinoplasty, there is a possibility of petroleum ointment migrating into intranasal incisions or osteotomy sites. A 15-year-old girl was examined 1 year after endoscopic sinus surgery combined with an open rhinoplasty had been performed at another institution. Physical examination showed a widened nasal dorsum with an open-roof deformity and a 1-cm firm mass lateral to her lateral nasal sidewall. The pathological examination after removal of the cyst showed a foreign-body inclusion cyst or lipogranuloma. The cyst contained a petroleum-based substance.

Adolescent↗

Uncoupling proteins 2 and 3 are highly active H(+) transporters and highly nucleotide sensitive when activated by coenzyme Q (ubiquinone).

Based on the discovery of coenzyme Q (CoQ) as an obligatory cofactor for H(+) transport by uncoupling protein 1 (UCP1) [Echtay, K. S., Winkler, E. & Klingenberg, M. (2000) Nature (London) 408, 609-613] we show here that UCP2 and UCP3 are also highly active H(+) transporters and require CoQ and fatty acid for H(+) transport, which is inhibited by low concentrations of nucleotides. CoQ is proposed to facilitate injection of H(+) from fatty acid into UCP. Human UCP2 and 3 expressed in Escherichia coli inclusion bodies are solubilized, and by exchange of sarcosyl against digitonin, nucleotide binding as measured with 2'-O-[5-(dimethylamino)naphthalene-1-sulfonyl]-GTP can be restored. After reconstitution into vesicles, Cl(-) but no H(+) are transported. The addition of CoQ initiates H(+) transport in conjunction with fatty acids. This increase is fully sensitive to nucleotides. The rates are as high as with reconstituted UCP1 from mitochondria. Maximum activity is at a molar ratio of 1:300 of CoQ:phospholipid. In UCP2 as in UCP1, ATP is a stronger inhibitor than ADP, but in UCP3 ADP inhibits more strongly than ATP. Thus UCP2 and UCP3 are regulated differently by nucleotides, in line with their different physiological contexts. These results confirm the regulation of UCP2 and UCP3 by the same factors CoQ, fatty acids, and nucleotides as UCP1. They supersede reports that UCP2 and UCP3 may not be H(+) transporters.

Animals↗

Outbreak of herpesviral conjunctivitis and respiratory disease in gouldian finches.

An outbreak of tracheitis, sinusitis, and conjunctivitis, originating in recently imported birds, caused high morbidity and mortality in a flock of finches in Central Illinois. Although several species were present, Gouldian finches (Erythrura [Chloebia] gouldiae) were most commonly and severely affected. Birds submitted for necropsy displayed microscopic lesions characteristic of herpesviral infection, including epithelial cytomegaly and karyomegaly with basophilic, intranuclear inclusion bodies in the nasopharynx, sinuses, trachea, parabronchi, conjunctiva, and occasionally the lacrimal gland or proximal proventricular glands. Viral particles consistent with herpesvirus were visualized within affected epithelial cells with electron microscopy. Based on a partial sequence of the viral DNA polymerase gene, this virus was found to be identical to a herpesvirus previously implicated in a similar outbreak in Canada and is most likely an alphaherpesvirus.

Animals↗

Developing an epitope-driven tuberculosis (TB) vaccine.

Epitope-driven vaccines are created from selected sub-sequences of proteins, or epitopes, derived by scanning the protein sequences of pathogens for patterns of amino acids that permit binding to human MHC molecules. We developed a prototype tuberculosis (TB) vaccine that contains epitopes derived by (1) EpiMer mapping of previously published secreted proteins derived from Mycobacterium tuberculosis (Mtb), and (2) EpiMatrix mapping of selected Mtb genome open reading frames (ORFs). Each of the epitopes contains at least three distinct class II MHC binding motif matches. These Mtb epitope selections were validated by measuring T cell responses from peripheral blood mononuclear cells (PBMC) obtained from healthy, asymptomatic tuberculin skin test-positive donors. Twenty-four validated Mtb epitopes were selected for inclusion in a DNA plasmid vector. We immunized HLA-DR B*0101 transgenic mice with this vaccine prototype augmented by co-administration of rIL-15. Following administration of three immunizations at 14-day intervals in conjunction with rIL-15, epitope-specific T cell responses were observed to eight of the 24 epitopes contained in the DNA construct, one week following the last injection. The systematic application of bioinformatics tools to whole genomes, in combination with in vitro methods for screening and confirming epitopes, may lead to the development of novel vaccines for infectious diseases like TB.

Amino Acid Sequence↗

Solitary fibrous tumor. Consistent CD34 immunoreactivity and occurrence in the orbit.

The distinction of solitary fibrous tumors (SFTs) from histologically similar neoplasms relies heavily on a characteristic microscopic appearance. No discriminating ultrastructural or immunohistochemical features are known. We evaluated 22 SFTs and 118 other tumors often considered in the differential diagnosis for immunoreactivity using a monoclonal antibody directed against the human hematopoietic progenitor cell antigen, CD34. All the SFTs (22 of 22, 100%) demonstrated strong CD34 immunoreactivity, irrespective of tumor site and histologic grade. Strong and generalized CD34 positivity was also found in most dermatofibrosarcoma protuberans (11 of 12, 92%) and occasional smooth-muscle tumors (leiomyomas 2 of 11, 18%; leiomyosarcomas 2 of 11, 18%). Variable numbers of CD34 positive cells were present in all neurofibromas (9 of 9, 100%) and focally present in most schwannomas (8 of 9, 89%). Some of the hemangiopericytomas (7 of 16, 44%) exhibited CD34 immunoreactivity, however, generally with weak intensity and patchy distribution. CD34 immunoreactivity was not observed in mesotheliomas (0 of 20, 0%), synovial sarcomas (0 of 13, 0%), fibrosarcomas (0 of 12, 0%), or spindle-cell thymomas (0 of 5, 0%). We conclude that CD34 immunoreactivity is a sensitive marker for SFT and, in conjunction with an appropriate immunohistochemical panel, may be useful in discriminating SFTs from other histologically similar neoplasms. The observation that some mesenchymal stromal cells and SFTs share a CD34-positive immunophenotype suggests a histogenetic relationship. The inclusion in this study of two cases of SFTs arising in the orbit establishes another site of origin for this tumor and provides further support for a mesenchymal histogenesis.

Antibodies, Monoclonal↗

Adenovirus infection of the cervix.

A case is recorded in which intranuclear inclusions suggesting adenovirus infection were recognized in a routine Papanicolaou smear. The patient was recalled for virologic studies and adenovirus type 19 was cultured from the cervix. The patient had a recent history of systemic illness with conjunctivitis, and a friend had conjunctivitis at the same time which was also proven to be due to adenovirus of the same serotype. This is the first reported instance in which adenovirus infection of the cervix suspected on cytologic findings has been confirmed by virus culture.

Adenoviridae Infections↗

Activation of the esophagin promoter during esophageal epithelial cell differentiation.

Esophagin is a member of the small proline-rich protein family of cell envelope precursor proteins, which are expressed during squamous cell differentiation. Esophagin is expressed at high levels in normal esophageal epithelium, but its expression is absent from esophageal squamous cell carcinomas and adenocarcinomas. Moreover, loss of esophagin expression is present in areas of dysplasia or normal mucosa adjacent to carcinomas, suggesting that absence of esophagin may constitute a harbinger of early esophageal malignant transformation. A greater understanding of transcriptional control of esophagin may provide valuable insights into esophageal malignancy. Therefore, this study was undertaken in order to isolate and carry out initial characterization of a functional promoter for esophagin. A genomic clone containing esophagin was isolated and sequenced, including 2.7 kb of the esophagin promoter region. Esophagin expression was studied in response to various treatments of primary cultured human esophageal epithelial cells and squamous cell carcinoma cell lines. Calcium was the strongest inducer of the endogenous esophagin promoter, with induction occurring at 12-72 hours. In primary cultured esophageal epithelial cells, a region spanning 116 bp upstream of the transcriptional start site to 45 bp downstream was sufficient to direct low, basal, in vitro esophagin expression. However, responsiveness of primary esophageal cells to calcium required inclusion of promoter elements 1688 bp upstream of the transcriptional start site. Site-directed mutagenesis studies suggested a putative role for C/EBP-beta, OCT-1, and OCT-3 transcription factor binding sites in the minimal promoter region. In conjunction with published human in vivo studies, these data support the hypothesis that esophagin is a biomarker of esophageal squamous cell differentiation and provide an in vitro model to evaluate regulatory factors involved in this differentiation process.

Epithelium↗