CT histogram analysis to distinguish between acute intracerebral hemorrhage and cavernous hemangioma.

OBJECTIVE: Acute intracerebral hemorrhage (AICH) and cerebral cavernous hemangioma (CCM) are two common cerebral hemorrhage diseases with partially overlapping CT findings and clinical symptoms, making it hard to distinguish between them. The current study used histogram analysis based on CT images to differentiate between CCM and AICH and test its diagnosis performance. METHODS: This retrospective study included 158 patients with CCM and 137 patients with AICH. The histograms of brain CT plain scan images of both groups were extracted using Python code and included 18 histogram parameters of the lesions. The most effective parameters were selected by univariate logistic regression analysis and Spearman correlation analysis and included in the final multivariate logistic regression model. The sample was randomly divided into the training set and the validation set by 7:3. The ROC curve was constructed to evaluate the discriminant efficiency of the final logistic regression model in distinguishing between AICH and CCM. RESULTS: The univariate analysis identified seven significant histogram parameters with the following final logistic regression model: F = 3.731 + 2.6411 × 10-9 × Energy-1.192 × Kurtosis-0.003 × Minimum-1.449 × Skewness + 2.5002 × 10-10 × Total Energy-1.103 × Uniformity+0.009 × Variance. The model showed good diagnostic performance in distinguishing between AICH and CCM, with an AUC of 0.876, sensitivity of 70.8%, and specificity of 91.9% in the training set, and an AUC of 0.870, sensitivity of 82.9%, and specificity of 85.1% in the validation set. CONCLUSIONS: The histogram analysis of brain CT images can be used as an auxiliary method to distinguish between AICH and CCM effectively.

[Prospective study of relationship between metabolic diseases and stroke in Jinchang Cohort].

Objective: To understand the incidence of stroke in the population of Jinchang Cohort and the relationship between metabolic diseases and stroke, and provide scientific evidence for the prevention and treatment of stroke in the population. Methods: The epidemiological investigation data and physical examination data of the 33 042 follow-up participants in Jinchang Cohort were collected for a prospective cohort study. Restricted cubic splines functions was used to analyze the dose-response relationship between metabolic indexes and the risk of stroke incidence. Results: 1) The incidence rate of stroke in Jinchang Cohort was 1.59%, and the standardized incidence rate was 3.99%. 2) Hypertension (male HR=2.20, female HR=4.45) and dyslipidemia (male HR=1.49, female HR=1.79) were the risk factors of stroke incidence in the population and diabetes had influence on the incidence of stroke only in the males (HR=1.79), while obesity had influence only in the females (HR=1.64). The more kinds of metabolic diseases, the higher risk of stroke incidence was. 3) Systolic blood pressure had a non-linear dose-response correlation with the risk of stroke incidence, while diastolic blood pressure had a positive linear correlation with the risk of stroke incidence. Conclusions: The incidence of stroke in Jinchang Cohort population was high compared with both domestic level and oversea level. The patients with metabolic diseases were the population at high-risk for stroke, and more attention should be paid to them in the prevention and treatment of stroke. Diastolic blood pressure might be more closely related to stroke.

[Risk factors analysis and a new risk scoring system predicting hepatocarcinogenesis for chronic genotype C HBV infected patients].

Objective: To investigate the effects of hepatitis B virus (HBV) genotype and mutations on the development of hepatocellular carcinoma (HCC) and to establish a new qualified HCC risk scores. Methods: A cohort study enrolling patients with chronic HBV infection was conducted. HBV genotypes were identified by nested multiplex PCR. HBV mutations in the basic core promoter region and PreS region were sequenced after PCR amplification. Scores on risk factors were set based on nomogram. Results: Totally, 1 525 patients were followed-up in this research. A total of 1 110 patients infected with genotype C were followed-up for 8.52 (Q(R): 5.36-11.68) years on average, of whom the incidence of HCC was 11.93/1 000 person-years. In genotype C HBV infected patients, male gender, aged 40 years and over, and four DNA mutations (T1674CG, A1762T/G1764A, A3120T, and A2962G) can increase the risk of HCC (P<0.05); interferon therapy can reduce the risk of HCC (P<0.05). A new HCC predicting model was established according to the results. After validation, the predicted disease-free survival rate was consistent with the real one. Conclusions: Hepatitis B virus genotypes and mutations were closely associated with HCC. The new risk scoring system can well predict HCC occurrence in genotype C HBV infected patients.

Pharmacovigilance in China: issues of concern identified through an analysis of the Chinese Adverse Drug Reaction Information Bulletin 2001 to 2014.

WHAT IS KNOWN AND OBJECTIVE: The Adverse Drug Reaction Information Bulletin (ADRIB), issued by China Food and Drug Administration (CFDA), is a major source of information on drugs causing safety concerns in China. As the publication is only published in Chinese, we undertook an analysis of the reports in the ADRIB since its first publication in 2001 to give international readers a better appreciation of the pharmacovigilance issues addressed. METHODS: Every issue of the ADRIB was scrutinized, and the issues addressed as well as the drugs involved are summarized and discussed. RESULTS AND DISCUSSION: From 2001 to 2014, 109 items of ADR information have been reported. The antimicrobial agents were most often the subject of discussion. There were 28 traditional Chinese medicines (TCMs) discussed. Among the ADRs addressed, the adverse reactions of the skin and its appendages were most frequent. About two-fifths of the ADRs arose from the inherent properties of the active substance, and a majority of the ADRs were caused by off-label use, irrational drug combinations and misuse in special populations. WHAT IS NEW AND CONCLUSION: Many of the pharmacovigilance issues addressed were similar to those considered by Western Drug Regulatory Agencies. The pharmacovigilance issues relating to Chinese traditional medicines are less well addressed internationally, and these would be of particular value as the use of such medicines increases in the West.

The comparisons of the efficacy and toxicity between gefitinib and docetaxel for patients with advanced nonsmall-cell lung cancer: a meta-analysis from randomized controlled clinical trials.

BACKGROUND: The extent of the benefit of gefitinib in the treatment of advanced nonsmall-cell lung cancer (NSCLC) is till controversial, when compared with docetaxel. We performed this meta-analysis to compare the efficacy and toxicity of gefitinib with docetaxel for different patients with advanced NSCLC. MATERIALS AND METHODS: We searched PubMed, Cochrane Library, and identified 5 randomized controlled clinical trials published within 2000-2013. After further full-text screening, 4 clinical trials were included in the final meta-analysis. RESULTS: The outcomes of treatment efficacy included progression-free survival (PFS), overall survival (OS) and objective response rate (ORR). Comparing gefitinib to docetaxel for advanced NSCLC patients, the pooled hazard ratio (HR) of PFS was 0.91, (95% confidential index [CI] = 0.83-0.99), the pooled HR of OS was 1.02, (95% CI = 0.93-1.13), the pooled risk ratio of ORR was 1.57, (95% CI = 1.01-2.47). CONCLUSIONS: Gefitinib was found to significantly improve patients' PFS and response rate compared with docetaxel. There is no difference of OS between gefitinib and docetaxel.

Cellular immune responses to feline immunodeficiency virus (FIV) induced by dual-subtype FIV vaccine.

Vaccine-induced T cell responses to FIV were assessed by measuring FIV-specific cytokine and cytotoxic-effector molecule production. A total of 22 cats at 10-12 weeks of age received either dual-subtype FIV vaccine (n=12), uninfected cell lysate (n=5) consisting of cells used to produce vaccine viruses, or no immunization (n=5). Significant increases in mRNA and protein production of T-helper 1 (Th1) cytokines (IL-2, IFNgamma), mRNA production of a cytotoxic-effector molecule (perforin), and lymphoproliferation response were observed in peripheral blood mononuclear cells (PBMC) from dual-subtype FIV-vaccinated cats after in vitro stimulation with inactivated FIV. In contrast, no statistically significant increase in FIV-stimulated mRNA production of Th2 cytokines (IL-4, IL-6) or other cytotoxic-effector molecules (TNFalpha, FasL) was observed in the PBMC from dual-subtype vaccinated cats. Moreover, no FIV-specific increases in the IFNgamma, IL-2, and perforin mRNA productions and in the IFNgamma bioactivity and lymphoproliferation responses were observed in the PBMC from cell-immunized cats. These observations suggest that IFNgamma induction, lymphoproliferation, and significant portion of IL-2 and perforin productions in the PBMC from dual-subtype vaccinated cats are clearly specific for viral antigens. Overall, dual-subtype FIV vaccine elicited strong Th1 response (IFN(, IL-2), which may contribute to the vaccine protection by enhancing the perforin-mediated cytotoxic-cell activity against FIV.

FIV vaccine development and its importance to veterinary and human medicine: a review FIV vaccine 2002 update and review.

Feline immunodeficiency virus (FIV) is a natural infection of domestic cats that results in acquired immunodeficiency syndrome resembling human immunodeficiency virus (HIV) infection in humans. The worldwide prevalence of FIV infection in domestic cats has been reported to range from 1 to 28%. Hence, an effective FIV vaccine will have an important impact on veterinary medicine in addition to being used as a small animal AIDS model for humans. Since the discovery of FIV reported in 1987, FIV vaccine research has pursued both molecular and conventional vaccine approaches toward the development of a commercial product. Published FIV vaccine trial results from 1998 to the present have been compiled to update the veterinary clinical and research communities on the immunologic and experimental efficacy status of these vaccines. A brief report is included on the outcome of the 10 years of collaborative work between industry and academia which led to recent USDA approval of the first animal lentivirus vaccine, the dual-subtype FIV vaccine. The immunogenicity and efficacy of the experimental prototype, dual-subtype FIV vaccine and the efficacy of the currently approved commercial, dual-subtype FIV vaccine (Fel-O-Vax FIV) are discussed. Potential cross-reactivity complications between commercial FIV diagnostic tests, Idexx Snap Combo Test and Western blot assays, and sera from previously vaccinated cats are also discussed. Finally, recommendations are made for unbiased critical testing of new FIV vaccines, the currently USDA approved vaccine, and future vaccines in development.

Dual-subtype FIV vaccine protects cats against in vivo swarms of both homologous and heterologous subtype FIV isolates.

OBJECTIVE: To evaluate the immunogenicity and efficacy of an inactivated dual-subtype feline immunodeficiency virus (FIV) vaccine. DESIGN: Specific-pathogen-free cats were immunized with dual-subtype (subtype A FIV(Pet) and subtype D FIV(Shi)) vaccine and challenged with either in vivo- or in vitro-derived FIV inocula. METHODS: Dual-subtype vaccinated, single-subtype vaccinated, and placebo-immunized cats were challenged within vivo-derived heterologous subtype B FIV(Bang) [10--100 50% cat infectious doses (CID(50))], in vivo-derived homologous FIV(Shi)(50 CID(50)), and in vitro- and in vivo-derived homologous FIV(Pet)(20--50 CID(50)). Dual-subtype vaccine immunogenicity and efficacy were evaluated and compared to single-subtype strain vaccines. FIV infection was determined using virus isolation and proviral PCR of peripheral blood mononuclear cells and lymphoid tissues. RESULTS: Four out of five dual-subtype vaccinated cats were protected against low-dose FIV(Bang) (10 CID(50)) and subsequently against in vivo-derived FIV(Pet) (50 CID(50)) challenge, whereas all placebo-immunized cats became infected. Furthermore, dual-subtype vaccine protected two out of five cats against high-dose FIV(Bang) challenge (100 CID(50)) which infected seven out of eight single-subtype vaccinated cats. All dual-subtype vaccinated cats were protected against in vivo-derived FIV(Pet), but only one out of five single-subtype vaccinated cats were protected against in vivo-derived FIV(Pet). Dual-subtype vaccination induced broad-spectrum virus-neutralizing antibodies and FIV-specific interferon-gamma responses along with elevated FIV-specific perforin mRNA levels, suggesting an increase in cytotoxic cell activities. CONCLUSION: Dual-subtype vaccinated cats developed broad-spectrum humoral and cellular immunity which protected cats against in vivo-derived inocula of homologous and heterologous FIV subtypes. Thus, multi-subtype antigen vaccines may be an effective strategy against AIDS viruses.

Immunohistochemical and in situ hybridization studies of gonadotropin releasing hormone (GnRH) and its receptor in rat digestive tract.

GnRH(LH-RH) is first discovered in the hypothalamus and found to have a role in regulation of reproduction. With the study on it deepening, GnRH was demonnstrated that it also exists in a number of organs beyond the hypothalamus and acts on extrapituitary organs. To study whether digestive tract synthesizes GnRH and its receptor and, if it does, by what cells. In the experiment, the locallizations of GnRH and its receptors in rat digestive tract were studied using immunohistochemistry and in situ hybridization. The parietal cells of gastric gland, the villous and glandular epithelium in small and large intestine and parasympathetic ganglion cells of myenteric plexus showed GnRH immunoreactivity; GnRH mRNA hybridization signal was detected. The epithelium of gastric pit and the cells above in digestive tract showed GnRH receptor immunoreactivity; GnRH receptor mRNA hybridization signal was detected. The immunoreactive and signal materials distributed in cytoplasm of all positive cells, with nuclei being immunonegative and with no hybridization signal. These results suggested that the digestive tract can produce GnRH and express GnRH receptor; GnRH may also be a gastrointestinal hormone.

Presence of gonadotropin-releasing hormone (GnRH) and its mRNA in rat pancreas.

Gonadotropin-releasing hormone (GnRH) has been reported to exist in many non-hypothalamic tissues, such as the placenta, gonads, and mammary glands, while there still have been no reports concerning the existence and expression of GnRH in the mammalian digestive system. Immunocytochemistry and in situ hybridization results show that GnRH molecule and GnRH mRNA are both exclusively distributed in exocrine pancreas, and RT-PCR result further proves that GnRH transcription units do exist in the pancreas, which possess the same sequence as the hypothalamus GnRH mRNA. Quantitative analysis indicates that mRNA levels in rat pancreas remain at a low level (less than 10% of that in hypothalamus) without sexual or developmental difference. This is the first report suggesting the existence and gene expression of GnRH in rat pancreas.

Cortical actin filaments form rapidly during photopolarization and are required for the development of calcium gradients in Pelvetia compressa zygotes.

Previous research has shown that cortical gradients of cytosolic Ca(2+) are formed during the photopolarization of Pelvetia compressa zygotes, with elevated Ca(2+) on the shaded hemisphere that will become the site of rhizoid germination. We report here that the marine sponge toxin, latrunculin B, which blocks photopolarization at nanomolar concentrations, inhibited the formation of the light-driven Ca(2+) gradients. Using low concentrations of microinjected fluorescent phalloidin as a tracer for actin filaments, we found that exposure to light induced a striking increase in actin filaments in the cells as indicated by an increase in fluorescence. The increase was quantified in the cortex, where it was most apparent, and the fluorescence there was found to increase by about a factor of 3. This increase in cortical phalloidin fluorescence was inhibited by latrunculin B at the same concentration required to inhibit Ca(2+) gradient formation and photopolarization. The distribution of the increasing phalloidin fluorescence was uniform with respect to the developing rhizoid-thallus axis during the formation of the axis, and no intense patches of fluorescence were observed. After germination, fluorescence suggestive of an apical ring of actin filaments was seen near the rhizoid tip. Finally, inhibitor studies indicated that myosin may be involved in the photopolarization process.

MHC-restricted protection of cats against FIV infection by adoptive transfer of immune cells from FIV-vaccinated donors.

The role of cellular immunity in vaccine protection against FIV infection was evaluated using adoptive cell transfer studies. Specific-pathogen-free cats received two adoptive transfers of washed blood cells from either vaccinated or unvaccinated donors with varying MHC compatibility at 1-week intervals, and a homologous FIV(Pet) challenge 1 day after the first adoptive transfer. FIV-specific CTL, IFN-gamma production, and proliferation responses were detected in the PBMC from the vaccinated donors. Seven of eleven (64%) recipients of cells from half-matched/vaccinated donors remained negative for FIV-antibodies after FIV challenge and four of those were completely protected. Two of two recipients of cells from MHC-identical/vaccinated donors were completely protected. All recipients of cells from unrelated/vaccinated, half-matched/unvaccinated, or unrelated/unvaccinated donors were unprotected. Thus, protection mediated by adoptive transfer of immunocytes from vaccinated cats was MHC-restricted, occurred in the absence of antiviral humoral immunity, and correlated with the transfer of cells with FIV-specific CTL and T-helper activities.

The ran GTPase regulates mitotic spindle assembly.

Ran is an abundant nuclear GTPase with a clear role in nuclear transport during interphase but with roles in mitotic regulation that are less well understood. The nucleotide-binding state of Ran is regulated by a GTPase activating protein, RanGAP1, and by a guanine nucleotide exchange factor, RCC1. Ran also interacts with a guanine nucleotide dissociation inhibitor, RanBP1. RanBP1 has a high affinity for GTP-bound Ran, and it acts as a cofactor for RanGAP1, increasing the rate of GAP-mediated GTP hydrolysis on Ran approximately tenfold. RanBP1 levels oscillate during the cell cycle [4], and increased concentrations of RanBP1 prolong mitosis in mammalian cells and in Xenopus egg extracts (our unpublished observations). We investigated how increased concentrations of RanBP1 disturb mitosis. We found that spindle assembly is dramatically disrupted when exogenous RanBP1 is added to M phase Xenopus egg extracts. We present evidence that the role of Ran in spindle assembly is independent of nuclear transport and is probably mediated through changes in microtubule dynamics.

Symmetry breaking in the zygotes of the fucoid algae: controversies and recent progress.

Despite its many advantages as an experimental system for the study of the epigenesis of polarity, it is obvious that the fucoid zygote also presents many problems. The development of polarity proceeds largely independently of direct gene action and thus may be considered a problem in cellular physiology. Ca2+ appears to play an important role in the process, but the optical properties of the zygotes (opacity and autofluorescence) hamper the use of modern methods of visualizing the distribution of Ca2+ and other ions. Likewise, other approaches, such as injection of fluorescent-labeled G-actin, in order to study the dynamics of actin filaments, are subject to the same limitations. It may be that the application of two-photon microscopy will enable experimenters to avoid some of these problems. This technique uses excitation wavelengths that are twice the wavelength of maximum absorption by fluorophores, and sufficient photon density for absorption is achieved only in a thin section. The fucoid zygotes are considerably more transparent to longer wavelengths, so attenuation of the exciting light and autofluorescence should be significantly reduced. Perhaps we will then be able to see further into these opaque cells. Another problem concerns the use of different species and genera. This may be unavoidable; for example, those of us who are land-locked tend to rely on Pelvetia, as it travels and stores better than the various species of Fucus and is less seasonal. Our colleagues fortunate enough to work near the ocean prefer to use the species that are locally available. Nevertheless, it is important to be careful about cross-genus and cross-species generalizations. While it is unlikely, based on what we know, that there are fundamental differences in physiological mechanisms among species, there may be small but still important differences in details. Obviously, investigators should directly compare results in more than one species whenever possible. The area of greatest disagreement, perhaps, concerns the mechanism of polarity formation, as opposed to its overt manifestation, germination. Are Ca2+ and actin involved or not? Assuming Ca2+ is involved, is the source internal or external? One basis for the different findings may be the differences in the strength of the polarizing signal provided to the zygotes. Clearly, the cells have powerful mechanisms for amplifying a faint asymmetry and developing an axis in response to an external signal. Furthermore, the fucoids generally develop in the intertidal zone and thus must be adapted to meeting the challenge of a widely varying external environment. They may have alternate mechanisms for responding to unilateral light. We have adopted the approach of presenting the cells with a fairly weak light signal--the minimum required to induce a considerable degree of organization of a population of zygotes. We then determine the effects of various inhibitors on photopolarization. One advantage of this approach is that it has allowed us to find treatments that increase the sensitivity of the zygotes to light, something that would not be possible if the untreated controls were fully polarized. Some of the differences between our results and those of others may be related to their use of a stronger light stimulus. It may be that if given a strong stimulus, a sufficient trace is left in the cells so that they can organize an axis when an inhibitor is removed. Careful consideration of this point may help to reconcile apparently contradictory findings. Despite these difficulties, the fucoid zygotes are likely to continue to be an important experimental system. Technology, including the development of more specific inhibitory reagents, may allow some of the shortcomings of the system to be overcome, and careful consideration of experimental conditions may resolve some of the points of disagreement.

Feline bone marrow transplantation: its use in FIV-infected cats.

The use of autologous and allogenic bone marrow transplantations (BMT) in FIV-infected and uninfected cats is a novel therapy for feline hematopoietic diseases and retroviral infections. A total of 13 specific pathogen-free (SPF) cats received either autologous or allogenic BMT and seven of these cats were also infected with FIV before autologous or allogenic BMT. All BMT recipients received total body irradiation of 900 cGy just before BMT. Two FIV-infected and four uninfected cats received autologous uninfected BM cells cryopreserved before BMT. Five infected and two uninfected cats received BM cells from allogenic uninfected donors (RBC-, MHC-, and cross-matched). MHC-matching was based on mixed leucocyte reaction (MLR) and the donor-recipient combination which was compatible by MLR analysis, was used in this study. Recipients were monitored for hematology, immunology, virology, and clinical signs. All FIV-infected and uninfected recipients of autologous BMT had complete engraftment with minimal complications. Uninfected recipients of allogenic BMT had a more severe clinical episode with slower rate of engraftment. None of these BMT groups had mortality. In contrast, only two of the five infected recipients of allogenic BMT survived for a significant period of time (23 and 50 weeks) and rest of the cats succumbed to transfusion reactions. Both infected BMT groups had persistent CD4/CD8 inversion, low CD4+ cell counts, and FIV infection of engrafted peripheral blood mononuclear cells (PBMC). Overall, successful autologous and allogenic BMTs were performed in FIV-free cats. All infected recipients of autologous BMT had compete engraftment and are currently alive, with thelongest survival time being over 1 year. Thus, BMT in combination with antiviral drug therapies may be an alternative therapy against retroviral infection.

Cytoplasmic calcium gradients and calmodulin in the early development of the fucoid alga Pelvetia compressa.

The predicted existence of cytoplasmic Ca2+ gradients during the photopolarization of the zygotes of the brown algae, Pelvetia and Fucus, has proved to be difficult to establish, and the downstream targets of the putative gradients are not known. We have used quantitative microinjection of the long excitation wavelength Ca2+ indicator, Calcium Crimson, and of antibodies against calmodulin to investigate these matters in the zygotes and early embryos of Pelvetia. We found that there is a window of cytoplasmic Calcium Crimson concentration that gives an adequate signal above autofluorescence yet allows normal development of the zygotes. As Calcium Crimson is not a ratiometric indicator, we injected other zygotes with a Ca2+-insensitive dye, rhodamine B, and imaged the cells at the same time that Calcium Crimson-injected cells were imaged. Ratios were calculated by dividing the averaged pixel values of Calcium Crimson images by the averaged pixel values of corresponding rhodamine B images. By this method, we observed the formation of a cytoplasmic Ca2+ gradient within one hour of the exposure of the cells to unilateral blue light during the photosensitive period. The region of high Ca2+ was localized to and predictive of the site of future rhizoid formation. We validated this somewhat indirect method by applying it to the growing rhizoid, where the existence of a tip-localized Ca2+ gradient is well established. The method clearly revealed the known gradient. The injection of ungerminated zygotes with antibodies made against Dictyostelium calmodulin inhibited germination, and this inhibition was abolished if the calmodulin antibodies were coinjected with an excess of purified maize calmodulin. Likewise, the growth of the rhizoids was inhibited by calmodulin antibody injections. The fungus-derived calmodulin antagonist, ophiobolin A, which has previously been shown to be a potent inhibitor of germination, also inhibited rhizoidal growth. Our results provide evidence that a cytoplasmic Ca2+ gradient is present during photopolarization and that calmodulin acts as a mediator of Ca2+ gradients throughout the early developmental processes of germination and rhizoidal growth in Pelvetia compressa.

Ubc9p and the conjugation of SUMO-1 to RanGAP1 and RanBP2.

The yeast UBC9 gene encodes a protein with homology to the E2 ubiquitin-conjugating enzymes that mediate the attachment of ubiquitin to substrate proteins [1]. Depletion of Ubc9p arrests cells in G2 or early M phase and stabilizes B-type cyclins [1]. p18(Ubc9), the Xenopus homolog of Ubc9p, associates specifically with p88(RanGAP1) and p340(RanBP2) [2]. Ran-binding protein 2 (p340(RanBP2)) is a nuclear pore protein [3] [4], and p88(RanGAP1) is a modified form of RanGAP1, a GTPase-activating protein for the small GTPase Ran [2]. It has recently been shown that mammalian RanGAP1 can be conjugated with SUMO-1, a small ubiquitin-related modifier [5-7], and that SUMO-1 conjugation promotes RanGAP1's interaction with RanBP2 [2,5,6]. Here we show that p18(Ubc9) acts as an E2-like enzyme for SUMO-1 conjugation, but not for ubiquitin conjugation. This suggests that the SUMO-1 conjugation pathway is biochemically similar to the ubiquitin conjugation pathway but uses a distinct set of enzymes and regulatory mechanisms. We also show that p18(Ubc9) interacts specifically with the internal repeat domain of RanBP2, which is a substrate for SUMO-1 conjugation in Xenopus egg extracts.

Efficacy evaluation of prime-boost protocol: canarypoxvirus-based feline immunodeficiency virus (FIV) vaccine and inactivated FIV-infected cell vaccine against heterologous FIV challenge in cats.

OBJECTIVE: To evaluate the immunogenicity and prophylactic efficacy of immunization schemes employing a recombinant canarypoxvirus ('ALVAC')-based feline immunodeficiency virus (FIV) vaccine alone or in combination with an inactivated FIV-infected cell vaccine against homologous and heterologous FIV challenges in cats. METHODS: Specific pathogen-free cats were given a total of three immunizations with subtype A vaccines and challenged 4 weeks after the final immunization with 50 median animal infectious doses (ID50) of FIV-Petaluma, a subtype A isolate. Following the initial challenge, protected cats received a second challenge with 75 ID50 of FIV-Bangston, a subtype B isolate. FIV-specific humoral and cell-mediated responses were measured to determine the immune correlates of protection. RESULTS: Two of three cats immunized with the ALVAC FIV recombinants alone were protected from homologous FIV challenge in the presence of FIV-specific cytotoxic T-lymphocyte (CTL) responses but in the absence of FIV-specific humoral responses. All three cats immunized with the ALVAC-FIV recombinant and boosted with FIV-infected cell vaccine were also protected from homologous FIV challenge in the presence of both FIV-specific CTL and humoral responses. Partial to full protection was observed in ALVAC-FIV/FIV-infected cell vaccine-immunized cats against a heterologous FIV challenge given 8 months after the initial challenge. Two out of three cats had transient infection and the remaining cat had no sign of FIV infection at a dose at which all three control cats were readily infected. CONCLUSIONS: Immunization schemes employing ALVAC-based FIV vaccines in combination with inactivated FIV-infected cell vaccine generate protective immune responses that can cross-react with FIV isolates that are genetically distinct from the vaccine strains.

The balance of RanBP1 and RCC1 is critical for nuclear assembly and nuclear transport.

Ran is a small GTPase that is essential for nuclear transport, mRNA processing, maintenance of structural integrity of nuclei, and cell cycle control. RanBP1 is a highly conserved Ran guanine nucleotide dissociation inhibitor. We sought to use Xenopus egg extracts for the development of an in vitro assay for RanBP1 activity in nuclear assembly, protein import, and DNA replication. Surprisingly, when we used anti-RanBP1 antibodies to immunodeplete RanBP1 from Xenopus egg extracts, we found that the extracts were also depleted of RCC1, Ran's guanine nucleotide exchange factor, suggesting that these proteins form a stable complex. In contrast to previous observations using extracts that had been depleted of RCC1 only, extracts lacking both RanBP1 and RCC1 (codepleted extracts) did not exhibit defects in assays of nuclear assembly, nuclear transport, or DNA replication. Addition of either recombinant RanBP1 or RCC1 to codepleted extracts to restore only one of the depleted proteins caused abnormal nuclear assembly and inhibited nuclear transport and DNA replication in a manner that could be rescued be further addition of RCC1 or RanBP1, respectively. Exogenous mutant Ran proteins could partially rescue nuclear function in extracts without RanBP1 or without RCC1, in a manner that was correlated with their nucleotide binding state. These results suggest that little RanBP1 or RCC1 is required for nuclear assembly, nuclear import, or DNA replication in the absence of the other protein. The results further suggest that the balance of GTP- and GDP-Ran is critical for proper nuclear assembly and function in vitro.