IL-24/MDA-7 Suppressed the Transplantation Tumor in BALB/c Mice

It is well-documented that interleukin-24 （IL-24） can induce apoptosis in a large spectrum of human cancer derived cell lines, but the effect of MDA-7/IL-24 gene transfer on mouse melanoma cells remains unknown. The eukaryotic expressing plasmid of IL-24 （pEGFP-IL-24） was constructed by DNA recombination technique. The recombination plasmid and empty vector were transfected into B16F0 cells and the expressions of IL-24 were determined by LSM, the proliferation of B16F0 cells was measured by MTT assay, and apoptosis rate and cell-cycle distribution of B16F0 cells were measured by FCM. The inhibitory effect of IL-24 gene transfection in mouse solid tumor was observed and measured. Compared with the control, the proliferation of B16F0 cells was inhibited by transfection with pEGFP-IL-24 and the G2/M phase of the transfected cells was also increased. Moreover, the percentage of mice with detectable tumor was decreased after inoculated with B16F0 cells transfected with pEGFP-IL-24. Growth rate of tumor in mouse model was significantly inhibited in IL-24 gene therapy group compared with the control. Proliferation of B16F0 cells was inhibited by pEGFP-IL-24 transfection. The intratumor injection of pEGFP-IL-24 could inhibit the growth of solid tumor in mice remarkably. Cellular ＆ Molecular Immunology.     

Pathways Contributing to Development of Spontaneous Mammary Tumors in BALB/c-Trp53+/− Mice

Mutation and loss of function in p53 are common features among human breast cancers. Here we use BALB/c-Trp53^+/− mice as a model to examine the sequence of events leading to mammary tumors. Mammary gland proliferation rates were similar in both BALB/c-Trp53^+/− mice and wild-type controls. In addition, sporadic mammary hyperplasias were rare in BALB/c-Trp53^+/− mice and not detectably different from those of wild-type controls. Among the 28 mammary tumors collected from BALB/c-Trp53^+/− mice, loss of heterozygosity for Trp53 was detected in more than 90% of invasive mammary tumors. Transplantation of Trp53^+/− ductal hyperplasias also indicated an association between loss of the wild-type allele of Trp53 and progression to invasive carcinomas. Therefore, loss of p53 function seems to be a rate-limiting step in progression. Moreover, expression of biomarkers such as estrogen receptor α, progesterone receptor, Her2/Neu, and activated Notch1 varied among mammary tumors, suggesting that multiple oncogenic lesions collaborate with loss of p53 function. Expression of biomarkers was retained when tumor fragments were transplanted to syngeneic hosts. Tumors expressing solely luminal or basal keratins were also observed (27 and 11%, respectively), but the largest class of tumors expressed both luminal and basal keratins (62%). Overall, this panel of transplantable tumors provides a resource for detailed evaluation of the cell lineages undergoing transformation and preclinical testing of therapeutic agents targeting a variety of oncogenic pathways including cancer stem cells.Compromised function of the p53 tumor suppressor pathway remains among the most common alterations found in cancers.^1,2 Although disruption of p53 function predisposes to a broad spectrum of malignancies, the breast epithelium seems exquisitely sensitive to proper p53 function. Polymorphisms in MDM2, CHK2, and ATM alter the stability and activity of p53 and have been linked to breast cancer risk in women. The activity of p53 has also been shown to be responsive to hormones in rodent models.^3–5 Exogenous estrogen and progesterone are sufficient to render the mammary epithelium resistant to carcinogen-induced tumors, mimicking the protective effect afforded by a full-term pregnancy,⁶ and the p53 pathway participates in the hormone-induced protection.^7,8 Furthermore, breast cancer is the most prevalent tumor among women with Li-Fraumeni syndrome, which is most commonly associated with heterozygous mutations in TP53.^9–11 Reduced dosage of the p53 gene has been associated with haploinsufficiency with respect to levels of p53 protein and activity, cell cycle arrest, apoptosis, and homology-directed DNA repair.^12–14 Therefore, the level of p53 activity is a critical regulator of tumor suppressor pathways and breast cancer risk.The mechanisms by which p53 suppresses tumors are diverse. The roles of p53 in mediating cell cycle arrest and apoptosis have been described extensively.¹⁵ More recently, the activities of p53 have been expanded to include regulation of DNA repair, senescence, autophagy, cellular metabolism, and microRNA processing.^16–21 In addition, loss of p53 was shown to permit expansion of the pool of pluripotent embryonic stem cells^22,23 and cancer stem cells.^23–25 The activities of p53 that are critical for suppression of tumors vary among tissues. In the thymus, the proapoptotic activity of p53 was necessary to suppress lymphomas, whereas the cell cycle checkpoint function was dispensable.²⁶ In contrast, senescence is the principal pathway leading to regression of liver tumors after restoration of p53 function²⁷ and seems to be the prominent pathway in sarcomas as well.²⁸ Among the known breast cancer susceptibility genes there is a convergence of function highlighting the central role of homology-directed repair of DNA double-strand breaks in breast cancer risk.²⁹ Thus, fidelity of double-strand break repair may be a critical pathway controlled by p53 in breast tissue.The sequential changes that occur in normal tissue and in premalignant mammary lesions as a consequence of heterozygous mutations in TP53 provide clues to the mechanisms that initiate the carcinogenic cascade as well as the cellular origins of breast cancer. Although it is difficult to monitor sequential changes in human breast cancers, spontaneous mammary tumors are common in BALB/c-Trp53^+/− female mice,^30,31 providing a model to examine the sequence of phenotypic and genetic alterations during mammary tumorigenesis. Using this model, we demonstrate that heterozygosity for Trp53 does not increase proliferation of the epithelium or the incidence of precancerous lesions. However, loss of the wild-type allele of Trp53 was associated with the transition from hyperplastic to invasive phenotypes. In a set of 28 spontaneous tumors, histological phenotypes and expression of oncogenes were heterogeneous. The majority of tumors expressed markers of both luminal and basal epithelia (62%), suggesting that progenitor cells are the most common origin. However, significant numbers of tumors expressed purely luminal keratins (27%) or basal keratins (11%). Although distinct patterns of keratins were observed, stem cell markers were expressed similarly in tumors with only keratins associated with luminal cells (K8/18) as well as tumors expressing both luminal and basal cell keratins (K8/18 and K5/6). Therefore, it seems that tumors arise most frequently from progenitor cells, which then commit toward more differentiated lineages during progression. Lineage decisions were not associated with specific activation of either Notch1 or Her2. Because tumor phenotypes were stable after transplantation, this panel of tumors can be used to speed preclinical testing of chemotherapeutic agents.     

Effect of Andrographis paniculata as an Adjuvant in Combined Chemo-Radio and Whole Body Hyperthermia Treatment—A Preliminary Study

Modulation of immune responses to alleviate disease has been of interest for a long time. Intraperitoneal administration of Andrographis paniculata extract along with whole body hyperthermia (WBH) was found to enhance the total WBC count in cyclophosphamide (CTX) and radiation treated animals when compared to untreated control animals. Maximum inhibition in the solid tumor development was observed when the CTX and radiation exposed animals were treated with extract in combination with whole-body hyperthermia. Similarly myeloperoxidase activity in tumor tissue from CTX and radiation-treated animals was also significantly inhibited when they were administered with Andrographis paniculata extract along with whole body hyperthermia. Moreover the production of cytokines such as IL-2 and GM-CSF, which was reduced after combined CTX and radiation treatment was significantly increased by the simultaneous treatment of extract and whole body hyperthermia. The elevated level of serum Tumor Necrosis Factor (TNF-α) level, after CTX and radiation treatment was also lowered significantly after the administration of extract and simultaneous exposure of whole-body hyperthermia with respect to untreated tumor-bearing animals.     

Augmentation of Natural Killer Cell and Antibody-Dependent Cellular Cytotoxicity in BALB/c Mice by Sulforaphane,a Naturally Occurring Isothiocyanate from Broccoli through Enhanced Production of Cytokines IL-2 and IFN-γ

Effect of sulforaphane on cell-mediated immune (CMI) response was studied in normal as well as Ehrlich ascites tumor-bearing BALB/c mice. Administration of sulforaphane significantly enhanced natural killer (NK) cell activity in both normal as well as tumor-bearing animals, and the activity was observed earlier than in tumor-bearing control animals. Antibody-dependent cellular cytotoxicity (ADCC) also was enhanced significantly in both normal as well as tumor-bearing animals after sulforaphane administration compared with untreated control tumor-bearing animals. An early antibody-dependent complement-mediated cytotoxicity (ACC) also was observed in sulforaphane-treated normal and tumor-bearing animals. Administration of sulforaphane significantly enhanced the production of Interleukin-2 and Interferon-γ in normal as well as tumor-bearing animals. In addition, sulforaphane significantly enhanced the proliferation of splenocytes, bone marrow cells, and thymocytes by stimulating the mitogenic potential of various mitogens such as concanavalin A, phytohaemagglutinin, poke weed mitogen, and lipopolysaccharide.     

Behavior of two Leishmania infantum strains—evaluation of susceptibility to antimonials and expression of microRNAs in experimentally infected J774 macrophages and in BALB/c mice

Parasitology Research - Strains of the same Leishmania parasite species, isolated from different host organisms, may exhibit unique infection profiles and induce a change in the expression of...     

False-Positive Rate of a “Fourth-Generation” HIV Antigen/Antibody Combination Assay in an Area of Low HIV Prevalence

We retrospectively analyzed the performance of the Architect HIV antigen/antibody (Ag/Ab) combination assay in a tertiary health care center with a situation of low HIV prevalence. The specificity and positive predictive value (PPV) were 99.78% and 31.21%, respectively. However, the specificity and PPV could increase to 99.99% and 89.70% using an arbitrary cutoff value.Since the first HIV test was introduced in 1985, the performances of HIV screening assays have improved continuously. In particular, the HIV antigen/antibody (Ag/Ab) combination assays launched in 1997 have shortened the window period by 4 to 5 days compared to those of the previous antibody-alone enzyme immunoassays (8). After the improvement of the detection limit of the p24 antigen enzyme immunoassay (EIA) to equivalent to that of the single-antigen EIA, HIV Ag/Ab combination assays have been implemented in numerous laboratories throughout the world (7, 9).The Centers for Disease Control and Prevention (CDC) recommended the expansion of HIV antibody testing from targeted, risk-based testing to universal screening of all adults aged 13 to 64 years in health care settings (2). Although the revised recommendation could be effective in identifying the maximum possible number of HIV-infected people from a public health perspective, there are concerns about false-positive results (3, 11). False-positive HIV screening results could cause substantial psychological distress while waiting for a confirmatory test (10). Guinn estimated the positive predictive value (PPV) of rapid HIV testing in Oregon with 100% sensitivity and 99.9% specificity to be 29% (3). The rate of false-positive results could be dramatically increased in situations of extremely low HIV prevalence. Shima-Sano et al. reported that the PPV of HIV screening results in pregnant women is only 3.7% (5).According to a report by the Korea Centers for Disease Control and Prevention, 6,120 individuals have been diagnosed with an HIV infection between 1985 and 2008 in Korea (4). Although the number of newly diagnosed HIV infections has increased, the cumulative number of HIV-infected individuals and prevalence were lower than those in other countries. There has been limited study on the rate of false-positive results in HIV screening tests using an automated HIV Ag/Ab combination assay. In the present study, we retrospectively analyzed the performance of an automated HIV Ag/Ab combination assay in a tertiary health care center with a situation of low HIV prevalence.During the period of 1 January 2006 through 31 December 2009, a total of 155,339 samples were tested for HIV using Architect HIV Ag/Ab Combo (HIV Combo; Abbott Laboratories, Abbott Park, IL) in a university hospital in South Korea. The Architect HIV Combo test was performed with an automated random access instrument (Architect i2000; Abbott Laboratories) throughout the study period. HIV Combo is a microparticle-based chemiluminescent immunoassay, designed for the simultaneous detection of HIV p24 antigen and HIV-1 and HIV-2 antibodies. Assay results were presented as ratios of specimen signals to the cutoff values (S/CO), where an S/CO ratio greater than or equal to 1.00 is considered reactive. The assays were performed according to the manufacturer''s directions.Specimens that initially tested reactive were retested in duplicate, and repeatedly reactive specimens were subjected to a secondary screening test and confirmatory test. The Vitros anti-HIV 1+2 assay on the Vitros ECiQ immunodiagnostic system (Ortho Clinical Diagnostics, Raritan, NJ) was used as a secondary screening test. Due to the low sample volume, secondary screening assays were performed with 403 specimens among 507 repeatedly reactive specimens. Repeatedly reactive specimens were confirmed by Western blotting (WB) (HIV Blot version 2.2; Genelabs Diagnostics, Singapore) at the Korean National Institute of Health, Seoul, South Korea. Statistical analysis was performed using SPSS version 12.0 for Windows (SPSS Inc., Chicago, IL). This study was approved by the institutional review board of Yonsei University Health System.A total of 155,339 HIV screening tests for 132,934 patients were performed during the past 4 years; the median patient age was 42.6 years (range, 0 months to 97 years). As shown in Table ​Table1,1, 543 (0.350%) specimens were found to be reactive using the initial HIV Combo screening assay and repeatedly reactive using the duplicated retesting. By review of the previous laboratory results, 36 specimens were collected from previously confirmed HIV-infected individuals and excluded from supplementary testing such as secondary screening testing or WB analysis. The HIV antibody was confirmed in 157 specimens by WB, and 346 specimens were concluded to have false-positive results, corresponding to a specificity of 99.78% (exact binomial 95% confidence interval [CI], 99.75% to 99.80%). However, PPVs were significantly different between genders, as follows: 49.84% (95% CI, 44.09% to 55.59%) for males and 2.53% (95% CI, 0.83% to 5.80%) for females. These results could be explained by the epidemiologic characteristics of HIV infection in South Korea, showing that approximately 92% of HIV-infected individuals were male.

TABLE 1.

Specificities and positive predictive values of the Architect HIV Ag/Ab combination assay^a

Enhancement of Antitumor Effect of Tegafur/Uracil （UFT） plus Leucovorin by Combined Treatment with Protein-Bound Polysaccharide, PSK, in Mouse Models

We evaluated the antitumor effect of combined therapy with tegafur/uracil （UFT） plus leucovorin （LV） （UFT/LV） and protein-bound polysaccharide, PSK, in three mouse models of transplantable tumors. UFT/LV showed antitumor effect against Meth A sarcoma, and the antitumor effect was enhanced when PSK given concomitantly. UFT/LV showed antitumor effect to Lewis lung carcinoma and PSK alone also showed antitumor effect at high dose, but a combination of UFT/LV and PSK resulted in no enhanced antitumor effect. Colon 26 carcinoma was weakly responsive to UFT/LV, and no enhancement of antitumor effect was found even PSK was used in combination. In conclusion, while the effect of PSK varies depending on tumor, combined use of UFT/LV and PSK may be expected to augment the antitumor effect.     

Allergen-specific immunoglobulin,histamine and T-cell responses induced by soybean glycinin and β-conglycinin in BALB/c mice of oral sensitisation

Glycinin and β-conglycinin are major soybean allergens involved in food hypersensitivity. However, the mechanism of immune responses induced by glycinin and β-conglycinin has not been fully understood. Balb/c mice were oral sensitised with different doses (0.1, 1.0 and 10 mg/day) of soybean glycinin and β-conglycinin for five weeks. Allergen-specific immunoglobulin (Ig), serum histamine and T-cell responses were tested to assess the allergenic activity of glycinin and β-conglycinin. Mice sensitised with 0.1 or 1.0 mg/day allergens induced high levels of specific IgE, IgG1 and serum histamine compared with mice treated with saline. Furthermore, specific T-cell proliferation and significant up-regulation of interleukin (IL)-4, IL-5 and interferon (IFN)-γ were observed in splenocytes from mice gavaged with 0.1 or 1.0 mg/day soybean proteins. Low doses of glycinin or β-conglycinin can induce allergic reactions in BALB/c mice, which might be associated with increased IgE and cytokine production.     

External application of NF-κB inhibitor DHMEQ suppresses development of atopic dermatitis-like lesions induced with DNCB/OX in BALB/c mice

Context: Dehydroxymethylepoxyquinomicin (DHMEQ) which is originally developed as an analog of antibiotic epoxyquinomicin C is a specific and potent inhibitor of NF-κB and has been shown to possess promising potential as an anti-inflammatory and anti-tumor agent.

Objective: This study examines DHMEQ’s effect on therapeutic potential for atopic dermatitis (AD)-like lesions.

Materials and methods: AD lesions were chronically induced by the repetitive and alternative application of 2,4-dinitrochlorobenzene (DNCB) and oxazolone (OX) on ears in BALB/c mice. The mice were then externally treated with DHMEQ ointment. Macroscopic and microscopic changes of the skin lesions were observed and recorded.

Results: DHMEQ inhibited ear swelling and relieved clinical symptoms of the AD-like lesions induced by DNCB/OX in BALB/c mice. Histopathology examination illustrated that it significantly decreased DNCB/OX-induced epidermal thickness, the infiltration of inflammatory cells, and the count of mast cell. The elevated level of immunoglobulin E (IgE) in serum and the mRNA levels of interferon γ (IFN-γ), interleukin 4 (IL-4) and IL-13 in the ear tissues, were also suppressed by DHMEQ.

Discussion and conclusion: This study indicated that DHMEQ would be useful for the treatment of AD.     

The Antiasthma Effect of Neonatal BCG Vaccination Does Not Depend on the Th17/Th1 but IL-17/IFN-�� Balance in a BALB/c Mouse Asthma Model

Objective

This study aimed to determine whether the protective effects of the Mycobacterium bovis Bacillus Calmette?CGuérin (BCG) vaccination on allergic asthma are associated with the T helper (Th) 17/Th1 balance in a murine asthma model.

Methods

BALB/c neonates were vaccinated with BCG on the first day after birth, sensitized with ovalbumin, and then challenged with allergen. The resulting airway inflammation and responsiveness were measured. The levels of IL-17 and interferon (IFN)-?? in BALF and ratio of Th17/Th1 were investigated.

Results

We found that although BCG neonatal vaccination inhibited airway hyperresponsiveness and inflammation following allergen challenge in a BALB/c mouse asthma model, reduced levels of Th2 cytokines were not observed. However, BCG neonatal vaccination reduced IL-17 production and increased IFN-?? production in both the bronchoalveolar lavage fluid and the lung lymphocytes in asthmatic mice.

Conclusion

The antiasthma effects of neonatal BCG vaccination reversed the IL-17/IFN-?? imbalance in a murine asthma model but did not depend on modifying the Th17/Th1 balance.     

Effect of T0901317 on Hepatic Proinflammatory Gene Expression in ApoE−/− Mice Fed a High-fat/high-cholesterol Diet

Objective In present study, we employed cDNA-based microarray technique to investigate the effect of a synthetic LXR ligand T0901317 on hepatic gene expression of proinflammatory cytokines in apolipoprotein E knockout mice fed an atherogenic diet. Methods and results Male 8-week-old apoE−/− mice were randomly divided into four groups, baseline group, vehicle group, prevention group and treatment group. All of the mice were fed a high-fat/high-cholesterol diet with or without LXR agonist T0901317 for 8 or 14 weeks. Gene array analysis found 17 atherosclerosis-related genes with a 2- to 8-fold difference in expression level between vehicle-treated group and T0901317-treated group. It induced mRNA expression of proinflammatory cytokine tumor necrosis factor (TNF), but inhibited gene expression of several other proinflammatory cytokines including interleukin (IL)-1α, IL-6, and IL-7 in the liver. C-reactive protein, TNF, matrix metalloproteinase-9, IL-1α, IL-6, and IL-7 were verified by real-time quantitative PCR. Next, enzyme-linked immunosorbent assay analyses showed up-regulation of TNFα levels and down-regulation of IL-α, IL-6, IL-7 levels in plasma sample. Conclusion The synthetic LXR agonist T0901317 has paradoxical roles in hepatic gene expression of proinflammatory cytokines in apoE−/− mice.     

Hepato-Renal Pathology in Pkd2ws25/− Mice,an Animal Model of Autosomal Dominant Polycystic Kidney Disease

Polycystic liver diseases, the most important of which are autosomal dominant and autosomal recessive polycystic kidney diseases, are incurable pathological conditions. Animal models that resemble human pathology in these diseases provide an opportunity to study the mechanisms of cystogenesis and to test potential treatments. Here we demonstrate that Pkd2^ws25/− mice, an animal model of autosomal dominant polycystic kidney disease, developed hepatic cysts. As assessed by micro-computed tomography scanning of intact livers and by light microscopy of hepatic tissue, hepatic cystic volumes increased from 12.82 ± 3.16% (5- to 8-month-old mice) to 21.58 ± 4.81% (9- to 12-month-old mice). Renal cystogenesis was more severe at early stages of disease: in 5- to 7-month-old mice, cystic volumes represented 40.67 ± 5.48% of kidney parenchyma, whereas in older mice cysts occupied 31.04 ± 1.88% of kidney parenchyma. Mild fibrosis occurred only in liver, and its degree was unchanged with age. Hepatic cysts were lined by single or multiple layers of squamous cholangiocytes. Cystic cholangiocyte cilia were short and malformed, whereas in renal cysts they appeared normal. In Pkd2^ws25/− mice, mitotic and apoptotic indices in both kidney and liver were increased compared with wild-type mice. In conclusion, Pkd2^ws25/− mice exhibit hepatorenal pathology resembling human autosomal dominant polycystic kidney disease and represent a useful model to study mechanisms of cystogenesis and to evaluate treatment options.The polycystic liver diseases are a group of genetic disorders that may occur alone or in combination with polycystic kidney disease. The polycystic liver diseases are characterized by the presence of hepatic cysts with or without hepatic fibrosis.^1–3 We recently proposed the term “cholangiociliopathies” for these conditions because they are linked to abnormalities in the function and/or structure of cholangiocyte primary cilia.^2,4 The cholangiociliopathies include autosomal dominant polycystic kidney disease (ADPKD), autosomal recessive polycystic kidney disease (ARPKD), Meckel–Gruber syndrome, Bardet–Biedl syndrome, nephronophthisis, and Joubert syndrome. The most common polycystic liver diseases are ARPKD and ADPKD.² Presently there is no effective treatment for polycystic liver diseases. The availability of animal models of these conditions provides the opportunity to assess the molecular mechanisms of hepatorenal cystogenesis and to evaluate potential therapeutic interventions. Ideally, these models should be genetically orthologous with and exhibit the phenotype of human polycystic liver diseases.³A number of animal models to study the mechanisms of cystogenesis and to evaluate potential therapeutic strategies for hepatorenal pathology have been described.⁵ A number of these models (ie, cpk,^6,7 bpk,^8,9 and kat¹⁰ mice, and Han:SPRD-cy¹¹ and PCK rats^12–14) are spontaneous mutants while others were generated by chemical mutagenesis (jcpk mice),^8,15 transgenic technologies (orpk¹⁶ and inv^17,18 mice), or gene specific targeting (Pkd1 and Pkd2 mice).^3,19–23 In several models (eg, cpk and bpk mice and the PCK rat), the hepatic and renal pathology resemble the human phenotype (ie, cellular origin of the hepatic and renal cysts, their localization, onset of disease, etc). Some models are genetically orthologous to their human counterparts (eg, the PCK rat and inv mice), while others display features of ADPKD or ARPKD, but so far no human gene syntenic to the gene mutated in animal models has been found. The majority of animal models that exhibit liver pathology (ie, cpk, bpk, jcpk, orpk, and inv mice, and PCK rat) are the models of ARPKD. To date, the creation of mouse models of ADPKD with a liver phenotype has been less successful. Pkd1 or Pkd2 heterozygote mice usually have no distinct hepatorenal cystic phenotype, while homozygotes with mutations in either Pkd1 or Pkd2 genes die in utero or perinatally.^3,23,24In recent reviews,^3,5,25,26 all available animal models to study renal and hepatic pathology have been carefully analyzed. Based on these analyses, it appears that the best models to test experimental therapies for potential treatment of hepatorenal ciliopathies are the bpk, cpk, and Pkd2^ws25/− mice, and the PCK rat.³ Both bpk and cpk mice resemble human ARPKD and are caused by mutations in genes (Bicc1 and Cys1, respectively) that do not have human genetic orthologs.¹⁵ In the PCK rat, the gene, Pkhd1, is orthologous to the human PKHD1.²⁷ We have recently described in detail the hepatic pathology of the PCK rat¹⁴ and have used this model extensively to understand the mechanisms of hepatic cystogenesis and to test possible therapies.²⁸The Pkd2^ws25/− mouse, an animal model of ADPKD, was generated by inducing two mutations in mouse homolog Pkd2−true null mutation (ws183, designated Pkd2⁻) and an unstable recombinant-sensitive allele (ws25; designated Pkd2^ws25).²⁰ Pkd2^ws25/− mice develop liver cysts slowly and progressively, and to date, appear to be one of the best models to study the mechanisms of hepatic cystogenesis in ADPKD. Wu and colleagues²⁰ examined to a very limited degree hepatic phenotype in 10- to 11-week-old Pkd2^ws25/− mice (ie, number of cysts present in affected livers) while describing the development of this animal model.In the present paper, we study in detail the liver and kidney pathology in Pkd2^ws25/− mice throughout the course of disease progression (at ages of 5 to 8 months and 9 to 12 months) with a focus on the cystic and fibrotic volumes over time, the rate of cell proliferation, apoptosis, and ciliary morphology.     

Protective efficacy and safety of Brucella melitensis 16MΔmucR against intraperitoneal and aerosol challenge in BALB/c mice

Brucellosis is a zoonosis of nearly worldwide distribution. Vaccination against this pathogen is an important control strategy to prevent the disease. Currently licensed vaccine strains used in animals are unacceptable for human use due to undesirable side effects and modest protection. Substantial progress has been made during the past 10 years toward the development of improved vaccines for brucellosis. In part, this has been achieved by the identification and characterization of live attenuated mutants that are safer in the host but still can stimulate an adequate immune response. In the present study, the identification and characterization of the mucR mutant (BMEI 1364) as a vaccine candidate for brucellosis was conducted. BALB/c mice were vaccinated intraperitoneally at a dose of 10(5) CFU with the mutant to evaluate safety and protective efficacy against intraperitoneal and aerosol challenge. All animals vaccinated with the vaccine candidate demonstrated a statistically significant degree of protection against both intraperitoneal and aerosol challenge. Safety was revealed by the absence of Brucella associated pathological changes, including splenomegaly, hepatomegaly, or granulomatous disease. These results suggest that the 16MΔmucR vaccine is safe, elicits a strong protective immunity, and should be considered as a promising vaccine candidate for human use.     

Anti-Dopamine Antibodies: Effects on Behavior in an “Open Field,” Pain Sensitivity, CNS Monoamine Content, and Functional Activity of Immunocytes in C57Bl/6 Mice

Single i.p. doses of anti-dopamine antibodies were given to C57Bl/6 mice. This resulted in inhibition of motor activity in a large proportion of the animals in the open field test, which lasted five days. Hyperalgesia, detected 1.5 h and 1 day after doses of antibody, was replaced by analgesia on day 5. There was a sharp reduction in the levels of dopamine and its metabolites in the cerebral cortex at 1 and 5 days; the serotonin level was increased 1 day after doses of antibody, and was significantly decreased at 5 days. There was no effect on cells of the immune system. The possible mechanisms of the neurotropic action of these antibodies are discussed.     

Analysis of Treatment Costs for HIV RNA Reductions and CD4 Increases for Darunavir Versus Other Antiretrovirals in Treatment-Experienced,HIV–Infected Patients

Abstract

Background: For patients with preexisting HIV drug resistance, a wide range of antiretrovirals are used, with differences in efficacy and cost. The additional cost per incremental 25 cell rise in CD4 count, or 0.5 log reduction in HIV RNA, was calculated for 8 antiretrovirals using pivotal clinical trials data. Method: For approved antiretrovirals in HIV therapy–experienced patients, 24–week efficacy (benefit over control in HIV RNA and CD4 count) was extracted from pivotal trials in published reports and compared with the additional treatment cost versus the control arm of each trial (2006 US wholesale acquisition costs). Treatment costs in the POWER trials were calculated directly from the treatment use database. Results: Data were available from 11 clinical trials in more than 4,000 antiretroviral treatment-experienced patients: Gilead 907 (TDF vs. placebo), TORO1/2 (T-20/OBR vs. OBR), RESIST–1/2 (TPV/r vs. control PI), BMS-045 (ATV/r vs. LPV/r), CONTEXT (fAPV/r vs. LPV/r), CAESAR (3TC vs. placebo), CNA3002 (ABC vs. placebo), and POWER 1/2 (DRV/r vs. control PI). Additional cost per 0.5 log reduction in HIV RNA was $152 for ritonavir–boosted darunavir (DRV/r), $4,453 for lamivudine (3TC), $4,274 for abacavir (ABC), $4,641 for tenofovir (TDF), and $13,217 for enfuvirtide (T-20). Cost per 25 cell rise in CD4 ranged from $132 for darunavir/r to $16,464 for T-20. Conclusion: There is a wide range of costs associated with efficacy improvements across the classes of antiretrovirals used for antiretroviral treatment–experienced patients. This analysis does not account for differences in toxicity, use of concomitant medications, or long-term adherence, which could also influence value assessments.     

Number of Treatment Cycles Influences Development of Cytotoxic T Cells in Metastatic Breast Cancer Patients – A Phase I/II Study

The influence of the number of apheresis-stimulation-infusion(s) cycles, and the time in culture before the infusion (one vs. two weeks), on the generation of tumor antigen-specific cytotoxic T-lymphocytes (CTL) was investigated in a phase I/II clinical adoptive immunotherapy trial. Two previously treated metastatic breast cancer patients with no evidence of disease, in complete remission (CR), were enrolled. Each apheretic peripheral blood mononuclear cell (PBMC) sample was stimulated twice with MUC-1 before infusion back into the patients. Killer T-cells responses against MUC-1-expressing MCF-7 (CTL), nonspecific natural killer (NK) and lymphokine-activated killer (LAK) target cell lines, as well as, cytokine production were measured before each infusion. Patients received 2 infusions per month for 4 months. There were no tumor recurrences or toxicity. CTL, NK and LAK cells, type 1 cytokine, gamma-interferon (G-INF), and CD4⁺ and CD8⁺ memory T-lymphocytes were initially generated, produced or induced, respectively, and then declined. The CTL, NK and LAK cells were only induced at the first infusion of the first month. Thus, maintaining PBMC in culture longer than the first infusion was of no benefit with regards to retaining functional killer T-cells. In conclusion, this study implies that one treatment is optimal.