Clinical and radiological spectrum of acquired inflammatory demyelinating diseases of the central nervous system in a tertiary care center.

Objectives: Demyelinating diseases of central nervous system (CNS) are a broad spectrum of conditions with autoimmune process against myelin. In a resource limited country like India, it is imperative to perform proper clinical evaluation, neuroimaging to differentiate among various categories of CNS demyelinating diseases to decide regarding further workup and treatment. The objective of our study was to determine clinical presentation, imaging findings, serology results, diagnosis, and treatment outcome of primary demyelinating disorders of CNS. Materials and Methods: In this prospective study, a total of 44 patients were enrolled over a period of 1 year. After proper evaluation, patients were categorized into different groups applying newer diagnostic criteria. Patients were treated with steroids, appropriate immunomodulatory therapy, and outcomes were analyzed. Results: The majority of cases were of neuromyelitis optica spectrum disorder (NMOSD) (45.5%) with an overall female-to-male ratio of 3.4:1 and mean age of presentation was 30.5 ± 11.15. Myelitis (52.3%) followed by optic neuritis (45.5%) was the most common initial presentation. The most common site of involvement on magnetic resonance imaging was the spinal cord (particularly the cervicodorsal cord). The majority showed good response to therapy (77.27%) and two patients did not survive. Conclusion: Higher disability observed among seropositive NMOSD patients warrants aggressive treatment during the first attack itself. It is important to suspect myelin oligodendrocyte glycoprotein antibody disease in patients with preceding viral infection. A good outcome in the majority is likely due to the availability of serological assays and aggressive immunomodulatory therapy.

Attenuation of a virulent Aeromonas hydrophila with novobiocin and pathogenic characterization of the novobiocin-resistant strain.

AIM: To determine whether novobiocin resistance strategy could be used to attenuate a virulent Aeromonas hydrophila AH11P strain and to characterize the growth and pathogenic differences between the novobiocin-resistant strain and its virulent parent strain AH11P. METHODS AND RESULTS: A novobiocin-resistant strain AH11NOVO was obtained from a virulent Aer. hydrophila strain AH11P through selection of resistance to novobiocin. AH11NOVO was found to be avirulent to channel catfish (Ictalurus punctatus), whereas AH11P was virulent. When AH11NOVO vaccinated channel catfish were challenged with AH11P at 14 days postvaccination, relative per cent of survival of vaccinated fish was 100%. The cell proliferation rate of AH11NOVO was found to be significantly (P < 0.05) less than that of AH11P. In vitro motility assay revealed that AH11NOVO was nonmotile, whereas AH11P was motile. AH11NOVO had significantly (P < 0.05) lower in vitro chemotactic response to catfish mucus than that of AH11P. Although the ability of AH11NOVO to attach catfish gill cells was similar to that of AH11P, the ability of AH11NOVO to invade catfish gill cells was significantly (P < 0.05) lower than that of AH11P. CONCLUSIONS: The novobiocin-resistant AH11NOVO is attenuated and different from its parent AH11P in pathogenicity. SIGNIFICANCE AND IMPACT OF THE STUDY: The significantly lower chemotactic response and invasion ability of AH11NOVO compared with that of its virulent parent strain AH11P might shed light on the pathogenesis of Aer. hydrophila.

Requirements for both Rac1 and Cdc42 in membrane ruffling and phagocytosis in leukocytes.

Specific pathways linking heterotrimeric G proteins and Fcgamma receptors to the actin-based cytoskeleton are poorly understood. To test a requirement for Rho family members in cytoskeletal events mediated by structurally diverse receptors in leukocytes, we transfected the full-length human chemotactic peptide receptor in RAW 264.7 cells and examined cytoskeletal alterations in response to the chemotactic peptide formyl-methionyl-leucyl-phenylalanine (FMLP), colony stimulating factor-1 (CSF-1), IgG-coated particles, and phorbol 12-myristate 13-acetate (PMA). Expression of Rac1 N17, Cdc42 N17, or the GAP domain of n-chimaerin inhibited cytoskeletal responses to FMLP and CSF-1, and blocked phagocytosis. Accumulation of F-actin- rich "phagocytic cups" was partially inhibited by expression of Rac1 N17 or Cdc42 N17. In contrast, PMA-induced ruffling was not inhibited by expression of Rac1 N17, but was blocked by expression of Cdc42 N17, indicating that cytoskeletal inhibition by these constructs was nonoverlapping. These results demonstrate differential requirements for Rho family GTPases in leukocyte motility, and indicate that both Rac1 and Cdc42 are required for Fcgamma receptor- mediated phagocytosis and for membrane ruffling mediated by structurally distinct receptors in macrophages.

Effect of recombinant fibroblast interferon and recombinant immune interferon on growth and the antigenic phenotype of multidrug-resistant human glioblastoma multiforme cells.

To study the effect of drug resistance on the response of stage IV astrocytomas to interferon, a human glioblastoma multiforme cell line, GBM-18, was transfected with an expression-vector plasmid containing a human multidrug resistance (MDR) gene (pHaMDR1/A), and clones surviving in colchicine were isolated. GBM-18 multidrug-resistant subclones displayed cross-resistance to other chemotherapeutic agents, including vincristine, doxorubicin, and dactinomycin. The multidrug-resistant phenotype was reversible when GBM-18 multidrug-resistant cells were cultured in colchicine and the calcium-channel blocker verapamil. The level of the MDR1 gene (also known as PGY1) message was increased in GBM-18 multidrug-resistant cells selected for increased resistance to colchicine, and this effect was not correlated with an amplification of the MDR1 gene. In both parental GBM-18 and GBM-18 multidrug-resistant cells, growth was suppressed to a greater degree when cultures were treated with the combination of fibroblast interferon (IFN-beta) and immune interferon (IFN-gamma). Parental cells and multidrug-resistant subclones varied in their de novo and/or interferon-modulated expression of HLA class I and class II antigens, a high-molecular-weight melanoma-associated antigen, and intercellular adhesion molecule 1 (ICAM-1). Of the antigens tested, ICAM-1 and HLA class I antigens were the most sensitive to enhanced expression induced by IFN-beta and IFN-gamma when used alone or in combination. The results of the present study indicate that multidrug-resistant human glioblastoma multiforme cells retain their increased sensitivity to the antiproliferative activity of the combination of IFN-beta plus IFN-gamma, and differences in antigenic phenotype are apparent in independent multidrug-resistant glioblastoma multiforme clones.

Cloning and expression in Escherichia coli of an esterase-coding gene from the oil-degrading bacterium Acinetobacter calcoaceticus RAG-1.

A putative esterase gene (est) from Acinetobacter calcoaceticus RAG-1 has been cloned into Escherichia coli. Esterase-positive clones exhibited high levels of esterase activity even in intact cells. In addition, expression of the est gene conferred on E. coli the ability to grow on simple triglycerides such as triacetin (TAC). The original esterase-positive plasmid pRA17 carried a 2.2-kb insert from a partial MboI digest of RAG-1 DNA, which gave a single band with RAG-1 DNA following Southern hybridization. By subcloning and sequencing the est gene was found to contain a sequence of 870 bp which could be translated to yield a protein of Mr 32,700. In support of the sequencing results was the finding that when pRA17 was expressed in minicells, a unique peptide of Mr 32,500 was identified. This peptide was not found in minicells transformed with esterase-negative plasmids, such as pRA176, which contained a Tn5 insertion in the est gene. The fact that the production of active esterase depended on the orientation of the est gene within the vector suggested that transcription proceeded from the tet promoter in pBR322.

Rat urate oxidase: cloning and structural analysis of the gene and 5'-flanking region.

The structural gene (UOX) encoding rat urate oxidase (UOX) spans at least 23 kb and is composed of eight exons and seven introns. All of the exon-intron splice junction sequences conformed to the GT/AG consensus established for eukaryotic genes. The transcription start point (tsp) was determined using S1-type nuclease protection riboprobe, and assigned to an adenine 54 nucleotides (nt) upstream of the ATG start codon. A 456-bp 5'-terminal fragment, starting at the ATG codon, carries a putative TATA (ATAAAA) sequence at -32, and two putative 'CAAT box' sequences at -62 and -71 bp upstream from the tsp. No sequence resembling 'GC' box hexanucleotides (GGGCGG or CCGCCC) was found. The structural features of the 5'-flanking region of the UOX gene are distinct from the 5'-flanking sequences of peroxisomal beta-oxidation system genes which contain one or more 'GC' box elements but lack TATA- and CAAT-like features [Osumi et al., J. Biol. Chem. 262 (1987) 8138-8143; Ishii et al., J. Biol. Chem. 262 (1987) 8144-8150]. The 5'-flanking region of the UOX gene reveals a sequence, TTAGTAATT at nt -276 from the tsp, which appears to be complementary to the underlined part of the liver-specific LF-B1/HNF-1 consensus sequence, GTTAATNATTAAC (where N = A, C, T, G or no nt).

Growth factor receptor gene and protein expressions in the human lens.

In this study the mRNAs encoding epidermal growth factor receptor (EGFR), basic fibroblast growth factor receptor (FGFR-2) and insulin-like growth factor receptor (IGFR-1) genes of the human normal lenses at ages varying from 0.5 to 72 years, were identified by semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR). Regulation of EGFR gene expression in the lens did not change with aging, and of FGFR-2 and IGFR-1 genes also remained unaltered up to age 53 years. However, expressions of FGFR-2 and IGFR-1 genes were decreased at ages above 60 years. EGFR, FGFR-2 and IGFR-1 proteins were detected by immunoblot analysis in the epithelial cell membranes of lens at age varying from 40 to 72 years. There was no detectable amount of EGFR protein in fiber cell membranes of the lens, and the levels of FGFR-2 and IGFR-1 proteins were much lower than those in the epithelial cell membranes. The low levels of these receptor proteins in the fiber cell membranes of lens, suggest their possible role in keeping the differentiated function of these unique transparent cells. The findings of the increased protein levels with age of EGFR with the appearance of some degradation products at age 48 years and higher, and the increased FGFR-2 protein at age 60 years and higher in the epithelial cell membranes of lens, were of interest. It appears that this could be a compensatory protective response of the lens to aging process for lifelong continuation of normal growth by proliferation and differentiation of its epithelial cells into new fiber cells in the germinative zone at the equatorial region. Thus, these results could provide a basis for further studies on growth factor receptor gene and protein regulations in the mechanism of lens aging and progression of age-related human cataract.

Synthesis and biological activity of some new furan quaternary salts.

A series of new N-(5-substituted 2-furfuryl)-N,N-dimethyl-N-aryloxyalkyl quaternary ammonium salts relating to general structure IV has been synthesized by reacting 5-substituted 2-(N,N-dimethylaminomethyl)furans IIa-d with appropriate aryloxyalkyl bromides III. The resulting compounds are tested for in vitro antimicrobial activity. A simpler synthesis of 5-nitro-2-(N,N-dimethylaminomethyl)furan (IId) involving the reduction of N,N-dimethyl-5-nitro-2-furamide (Ib) with diborane is described. A new compound, 5-bromo-2-(N,N-dimethylaminomethyl)furnan (IIc), is prepared in a similar way. Many of these compounds (22, 28, 34, 37-42, 44, and 45) indicate high activity against Staphylococcus aureus, Streptococcus faecalis, Klebsiella pneumoniae, and Pseudomonas aeruginosa and are more active than nitrofurantoin, Compounds 22, 34 and 41 exhibit the highest in vitro antibacterial activity in the series. Some of these quaternary salts (22, 25, 37, 37-41, and 60) possess appreciable activity against Mycobacterium tuberculosis H37Rv. None of these compounds show significant antifungal activity. Eight compounds (18, 21, 22, 26-28, 32, and 34) have high in vitro antibacterial activity were inactive when tested for anthelmintic activity in rats against Nippostrongylus brasiliensis and Hymenolepis nana.

Novel euglycemic and hypolipidemic agents. 1.

A series of [[(heterocyclyl)ethoxy]benzyl]-2,4-thiazolidinediones have been synthesized by the condensation of corresponding aldehyde 1 and 2,4-thiazolidinedione followed by hydrogenation. Both unsaturated thiazolidinedione 2 and its saturated counterpart 3 have shown antihyperglycemic activity. Many of these compounds have shown superior euglycemic and hypolipidemic activity compared to troglitazone (CS 045). The indole analogue DRF-2189 (3g) was found to be a very potent insulin sensitizer, comparable to BRL-49653 in genetically obese C57BL/6J-ob/ob and 57BL/KsJ-db/db mice. Pharmacokinetic and tissue distribution studies conducted on BRL-49653 and DRF-2189 (3g) indicate that these drugs are well-distributed in target tissues. On the basis of euglycemic activity as well as enhanced selectivity against reduction of triglycerides in plasma, DRF-2189 (3g) has been selected for further evaluation.

Induction of immune response to bovine herpesvirus-1 with anti-idiotypic antibodies.

Previously, we prepared rabbit anti-idiotypic (anti-Id) antibodies against murine monoclonal antibodies (MAbs) specific for the major bovine herpesvirus-1 (BHV-1) envelope glycoproteins. Glycoprotein III (gIII) contains neutralization epitopes and may be the virus attachment protein. Anti-Id antibodies to a neutralizing MAb that reacts with gIII were purified by sequential immunoaffinity chromatography. Immune responses to the purified anti-Id reagent and BHV-1 were compared in mice. Both groups of mice produced BHV-1-specific neutralizing antibodies. However, lymphocyte proliferative responses and interferon and interleukin-2 production were specific for the respective immunizing antigens. These results suggest that the anti-Id reagent may bear an internal image of a B-cell-stimulating epitope of glycoprotein gIII; however, this epitope does not stimulate a virus-specific cellular immune response in mice.

Effects of bovine respiratory disease viruses and isoprinosine on bovine leukocyte function in vitro.

Peripheral blood mononuclear cells obtained from 4- to 6-month-old-calves were inoculated in vitro with bovine herpesvirus-1, parainfluenza-3, or bovine virus diarrhea viruses. No increase in infectious virus progeny was observed; however, the viruses were detected in the cells for at least 96 h post-infection without any significant reduction in cell viability. The three viruses, either alone or in combination, suppressed phytohemagglutinin-induced proliferation of the mononuclear cells. The greatest suppression was observed in cultures inoculated with bovine virus diarrhea virus. Addition of isoprinosine partially restored this viral-induced suppression of proliferative response, and the efficiency of reversal was greater in bovine virus diarrhea virus-infected cells. Interleukin-2 activity was higher in cultures of virus-infected mononuclear cells than in cultures of non-infected cells.

Effect of bovine herpesvirus-1 on expression of interleukin-2 receptors and effect of interleukin-12 on lymphocyte proliferation.

Expression of the interleukin-2 receptor alpha chain (IL-2R alpha) by peripheral blood mononuclear cells (PBMC) from Holstein calves, both experimentally-infected with bovine herpesvirus-l (BHV-l) and controls, was measured by flow cytometry. Expression of IL-2R alpha was 35 percent and 23 percent higher in infected calves than controls, on days 2 and 3 postinfection (PI), respectively. Concurrent with this increase in IL-2R alpha expression, a significant decrease (P < 0.001) was observed in the PHA-induced proliferative responses of PBMC from infected compared with control calves. In vitro treatment with recombinant human (rhu) IL-12 enhanced PHA-induced proliferative responses of PBMC from both infected and control calves. This rhuIL-12 enhancement of mitogen-induced proliferative responses was significant (P < 0.001) in infected calves on day 2 PI and was sufficient to abrogate the decrease observed due to BHV-1 infection. Since the expression of the beta and gamma chains of IL-2R was not measured it is difficult to speculate as to the status of high affinity receptor expression during BHV-1 infection. However results of the present study suggest that the decrease in proliferative responses observed during infection may not be due to a decrease in IL-2R alpha expression but may possibly be due to a selective down-regulation of signal transduction through IL-2R and/or by modulation of the expression of other cytokines involved in lymphocyte activation and proliferation.

Isolation and characterization of monoclonal antibodies to recombinant bovine interleukin-1 beta.

We describe the preparation of monoclonal antibodies (Mabs) directed against recombinant bovine interleukin-1 beta (rBoIL-1 beta). These anti-IL-1 beta Mabs were designated SA10, SA12, SA13, SA15, and SA22, and were characterized on the basis of their epitope specificity and cross-reactivity with homologous and heterologous cytokines in enzyme-linked immunosorbent assays and immunoblot analyses. Additionally, the ability of these Mabs to neutralize IL-1 beta was tested in thymocyte costimulation assays. The ELISA titers of all Mabs ranged from 9.4 x 10(6) to 1 x 10(7). Data indicate that Mabs SA10, SA12, SA15, and SA22 neutralized both bovine macrophage-derived IL-1 (1:4) and rBoIL-1 beta (1 ng ml-1). All the Mabs against rBoIL-1 beta (SA10, SA12, SA13, SA15, SA22) were specific and did not cross-react with other cytokines tested, except recombinant human IL-1 beta (rHuIL-1 beta). This finding suggests that these Mabs recognize epitopes common to human and bovine IL-1 molecules. Competition experiments suggested that Mab SA22 recognized a different epitope and Mabs SA10, SA12, SA13, and SA15 recognized the same epitope on the rBoIL-1 beta molecule. These observations suggest that these Mabs could be useful reagents for developing immunoassays to measure bovine IL-1 beta from biological fluids and to study the immunoregulatory role of IL-1 in the bovine immune system.

Changes in the leukocyte phenotype profile of goats infected with the caprine arthritis encephalitis virus.

The proportions of different sub-populations of leukocytes in five healthy goats and five goats infected with the caprine arthritis encephalitis virus (CAEV) were examined using immunofluorescence and flow cytometry. A panel of monoclonal antibodies that identified a monocytegranulocyte marker (GMI); the CD4, CD8, IgM, MHC Class I, MHC Class II and T19 antigens, and the gamma delta (gamma delta) T cell receptor was used. We observed a significant (P = 0.016) reduction in the proportion of monocytes in the peripheral blood of infected (5.98%) compared with healthy control goats (9.92%). There was also a decrease in the proportion of CD4+ T lymphocytes that approached significance (P = 0.076) accompanied by a slight increase in the proportion of CD8+ T lymphocytes, in infected compared with uninfected animals. Consequently, three of the five infected animals had lower CD4:CD8 ratios than any of the healthy animals and two of these three ratios were inverted. Approximately 14% of T cells in the peripheral blood of healthy goats was identified as gamma delta T cells and all expressed the T19 antigen. A significantly elevated level of gamma delta T cells (P = 0.030) and an elevated level of T19 cells were observed in infected, compared with healthy animals. The proportion of leukocytes expressing surface IgM (B cells) was also elevated, although not significantly, in CAEV-infected compared to healthy controls. The changes in peripheral blood leukocyte subsets in infected goats suggest that immune responses to the infection are probably altered in these animals with eventual progression to severe disease and death.

Immunopotentiation of bovine respiratory disease virus vaccines by interleukin-1 beta and interleukin-2.

Three experiments, using 85 crossbred beef calves, were conducted to evaluate the adjuvanticity of single, multiple, and combined doses of recombinant bovine IL-1 beta (rBoIL-1 beta) and recombinant bovine IL-2 (rBoIL-2), with a modified-live bovine herpesvirus-1/parainfluenza-3 (BHV-1/PI-3) virus vaccine and a killed bovine viral diarrhea (BVD) virus vaccine. Cytokines were administered intramuscularly at vaccination but at different injection sites. All cytokine treatments increased non-major histocompatibility complex (MHC)-restricted cytolytic capability of peripheral blood mononuclear cells (PBMC) against virus-infected target cells and serum neutralizing (SN) antibody titers to BHV-1 and BVD virus. Multiple, consecutive injections of rBoIL-2 generally showed the greatest adjuvant effect, and no additive effect was observed when rBoIL-1 beta and rBoIL-2 were administered together. In a challenge experiment, calves were vaccinated with a modified-live BHV-1/PI-3 vaccine and infected with BHV-1 on Day 21. Cytokine-treated calves had higher SN antibody titers to BHV-1 than did the control calves at the time of challenge. Calves that were administered rBoIL-2 on 5 consecutive days shed less BHV-1 and had the highest SN antibody titer to BHV-1 (Day 28). These data suggest that rBoIL-1 beta and rBoIL-2 may be useful immunoadjuvants for bovine respiratory disease virus vaccines.

Bovine recombinant interleukin-2 augments immunity and resistance to bovine herpesvirus infection.

The in vivo administration of bovine recombinant interleukin-2 (rIL-2) was evaluated in calves vaccinated and then challenged with bovine herpesvirus-1 (BHV-1). In Experiment 1, 24 calves were allotted to four groups: control; bovine rIL-2; BHV-1 vaccine (modified-live); and bovine rIL-2 + BHV-1 vaccine. Serum neutralizing antibody titers to BHV-1 were increased sixfold, and virus shedding was fourfold less in calves vaccinated and treated with rIL-2 (25 micrograms/kg, intramuscularly) when compared to calves that received vaccine only. Treatment with rIL-2 induced lymphokine-activated killer activity that was eliminated by pretreating effector cells with complement and a monoclonal antibody (B26A) specific for the sheep red blood cell receptor. The rIL-2 treatment in BHV-1-vaccinated calves increased the calves' ability to withstand a BHV-1 challenge. However, during treatment with rIL-2, calves developed diarrhea and mild fever that abated after IL-2 treatment was stopped. A second experiment was then conducted to determine a dose of rIL-2 that would enhance immunity to BHV-1 without causing adverse side effects. Twenty-five calves were allotted to five groups that received injections of rIL-2 at 0.0, 25.0, 2.5, 0.25, or 0.025 micrograms kg-1 day-1 for 5 days. All calves received a modified-live BHV-1 vaccine. Calves treated with 25.0 micrograms kg-1 day-1 showed similar adverse side effects as in the first experiment but all other calves were normal. Compared to control calves, those treated with 25.0, 2.5, and 0.25 micrograms kg-1 day-1 of rIL-2 had higher (P less than 0.05) serum antibody titers to BHV-1 and following challenge lower (P less than 0.05) BHV-1 titers in nasal secretions; additionally, clinical disease as evidenced by nasal and ocular discharge was less severe (P less than 0.05). In vitro cytotoxic responses against BHV-1-infected bovine kidney cells were increased (P less than 0.05) in calves treated with rIL-2 in a dose dependent manner. These data suggest that bovine rIL-2 at 2.5 to 0.25 micrograms/kg may be an effective adjuvant to immunization.

Effect of stress on interleukin-2 receptor expression by bovine mononuclear leukocytes.

Holstein calves given three consecutive i.m. injections of dexamethasone (DEX) (0.04 mg kg-1) showed lymphopenia and neutrophilia with increased numbers of mature neutrophils on post-injection Days 1 and 2, but these values returned to normal levels by post-injection Day 3. Interleukin-2 receptor (IL-2R) expression by peripheral blood mononuclear cells (PBMC) was evaluated by flow cytometry using a monoclonal antibody specific for bovine IL-2R alpha. Treatment with DEX significantly decreased expression of IL-2R alpha in Concanavalin A (Con A)-activated PBMC on Day 1 (P < 0.02) and on Day 2 (P < 0.1). On Day 3, expression of IL-2R alpha by PBMC was similar in control and DEX-treated calves. This decrease in IL-2R alpha expression correlated with decreased proliferative responses of PBMC to the T-cell mitogens, phytohemagglutinin (PHA) and Con A. Following in vitro treatment with recombinant human (rhu) interleukin-12 (IL-12) Con A-induced proliferative responses of PBMC tended to be higher in both groups. However, the rhu IL-12 induced increase of Con A activated proliferative responses were significantly greater in DEX-treated calves than in control calves. IL-2R alpha expression by PBMC was found to be less in calves transported 800 km in a truck as compared to that in PBMC from controls. These data suggest that stress-induced immunosuppression in calves may involve decreased IL-2R alpha expression and decreased IL-12 production. Serum chemistry results indicated a trend toward higher creatine kinase (CK) levels in DEX-treated calves. This may be due to the lysis of corticosteroid sensitive lymphocytes.

Facial nerve rehabilitation after radical parotidectomy.

OBJECTIVE: Examine functional outcomes in patients undergoing radical parotidectomy and facial nerve grafting. Identify factors that may affect rehabilitation in these patients. STUDY DESIGN: Retrospective chart review and photographic analyses of 12 patients undergoing radical parotidectomy with interposition nerve grafts for facial nerve reconstruction. METHODS: Data obtained for each patient regarding age, sex, histology of parotid neoplasm, cable graft source, administration of postoperative radiotherapy, and treatment for eye rehabilitation. Functional outcomes were assessed with the House-Brackmann grading system at 6 months, 1 year, and 2 years after surgery. RESULTS: All nerve grafts were harvested from cervical plexus sensory nerves with microscopic epineural repair performed for all neurorrhaphies. Overall, 9 of 12 patients achieved a grade III 2 years after surgery. All patients under age 30 obtained a grade III. Of the seven patients receiving postoperative radiation, five achieved a grade III. Older patients often required surgical procedures to facilitate eye closure. CONCLUSIONS: Facial nerve rehabilitation after radical parotidectomy can be successfully achieved with cervical plexus interposition nerve grafts. Postoperative radiotherapy did not appear to affect return of function, and younger patients consistently achieved good functional outcomes after nerve grafting. Older patients frequently require surgical procedures for eye rehabilitation after radical parotidectomy.

Comparison of culture, multiplex, and 5' nuclease polymerase chain reaction assays for the rapid detection of Yersinia enterocolitica in swine and pork products.

Bacteriological culture was compared with multiplex and fluorogenic (TaqMan) polymerase chain reaction (PCR) assays for the detection of attachment invasion locus (ail)-bearing Yersinia enterocolitica in market weight swine, chitterlings, and ground pork. The TaqMan assay detected 1 pg of purified Y. enterocolitica DNA, whereas conventional gel-based PCR detected I ng of the same. The presence of ail-bearing Y. enterocolitica was tested in pork and feces artificially inoculated with Y. enterocolitica strain NADC 5561. The sensitivity limits of culture, multiplex, and TaqMan PCR assays were 4 x 10(3), 4 x 10(2), and 0.4 CFU/g, respectively, for the artificially inoculated pork. The sensitivity limits were 4 x 10(2), 4 x 10(2), and 0.4 CFU/g, respectively, for feces after a 48-h enrichment in a Yersinia selective broth. By the culture method, Y. enterocolitica was not detected in any of the swine specimens (n = 2,403) examined. By contrast, it was detected in 48 (2%) of the swine samples screened using the multiplex PCR and in 656 (27.2%) of these samples using the TaqMan assay. Using the culture method, Y. enterocolitica was detected in 8% of chitterling samples (n = 350) and in none of the ground pork samples (n = 350). It was identified in 27% of the chitterling samples using multiplex PCR and in 79% of these samples using the TaqMan assay. Ten percent of the ground pork samples contained Y. enterocolitica, as determined by the multiplex PCR, and 38% based on the TaqMan assay. The results suggest that pork products harbor more ail-bearing Y. enterocolitica than selected organs of freshly slaughtered hogs and that the TaqMan assay is more sensitive than either the multiplex PCR or traditional culture methods.

Measurement of peripheral vestibular blood flow in a gerbil model of endolymphatic hydrops.

Presently, many investigators believe that the dysfunction of microcirculatory mechanisms may be responsible for vestibular symptoms in Meniere's disease. This study, using intravital microscopy (IVM), a technique that provides in vivo microcirculatory measures, was designed to determine whether impaired vestibular blood flow exists in endolymphatic hydrops. Hydrops was induced in the gerbil model by obliteration of the vestibular aqueduct and was confirmed histologically after IVM. Posthydrops gerbil subjects at 8 weeks as well as control animals were prepared for IVM. With a customized intravital microscope, red blood cell velocity and vessel diameter measurements were calculated for individual arterioles and capillaries from the microvascular bed at the horizontal ampulla. Mean arteriole diameter was significantly larger in the control group than in the hydrops group, whereas mean capillary diameters were similar for both groups. No significant difference was observed for mean red blood cell velocity in capillaries or arterioles between control and hydrops animals.