Upregulation of hepatic LDL receptor-related protein in nephrotic syndrome: response to statin therapy

Nephrotic syndrome (N-S) is associated with elevated plasma concentration and impaired clearance of VLDL, chylomicrons (CM), and their atherogenic remnants. These abnormalities are largely due to lipoprotein lipase, hepatic triglyceride lipase, and VLDL receptor deficiencies and impaired HDL-mediated shuttling of apoE and apoC between the nascent and remnant VLDL and CM. LRP is a multifaceted endocytic receptor that is heavily expressed in the liver. LRP recognizes at least 30 different ligands including VLDL and CM remnants. These observations prompted the present study to discern the effect of N-S on hepatic LRP gene and protein expressions. The study further sought to explore the effect of lipid-lowering therapy on LRP expression in N-S. Sprague-Dawley rats were randomized to the N-S (given ip injections of puromycin aminonucleoside; 130 mg/kg on day 1, 60 mg/kg on day 14) and placebo-injected control groups. On day 14, animals were subdivided into statin-treated (rosuvastatin; 20 mg x kg(-1) x day(-1) mixed with powdered chow) and untreated groups and studied on day 28. The untreated N-S group exhibited severe proteinuria, hypoalbuminemia, hypercholesterolemia, hypertriglyceridemia, and marked elevation of hepatic tissue LRP mRNA and protein abundance. Statin administration for 2 wk resulted in significant improvements of plasma lipid profile, proteinuria, and hypoalbuminemia as well as hepatic LRP mRNA and protein abundance. In contrast, statin administration had no significant effect on either plasma lipids or hepatic LRP levels in the normal control rats. In conclusion, N-S results in marked upregulation of hepatic LRP expression that is partly reversed with statin administration. These findings exclude depressed hepatic LRP expression as the primary cause of elevated plasma lipoprotein remnants in N-S.     

Tuberculosis masquerading as malignancy: a multimodality approach to the correct diagnosis – a case report

BACKGROUND: Extrapulmonary tuberculosis is one of the great mimickers of medicine, and often masquerades as malignancy. As a result, patients may be referred to oncologists and surgeons for further evaluation and management, delaying the institution of appropriate anti-tuberculous drug therapy. CASE PRESENTATION: We present the case of a 21 year old man with tuberculous osteomyelitis, who was referred to the Bone and Soft Tissue Sarcoma Service at our institution with a provisional diagnosis of malignancy. Further investigation revealed extensive retroperitoneal abdominal and pelvic lymphadenopathy. The recognition of certain patterns on imaging, and finally the isolation of Mycobacterium tuberculosis from tissue samples obtained under image guidance, enabled the correct diagnosis to be made. CONCLUSION: This case highlights the importance of remaining cognisant of the protean manifestations of extrapulmonary tuberculosis, and illustrates the advantage of a clinically directed multi-modality imaging approach to diagnosis.     

Comparison of Ionized Calcium-binding Adapter Molecule 1-Immunoreactive Microglia in the Spinal Cord Between Young Adult and Aged Dogs

Microglia are main form of active immune defense, and they are constantly moving and analyzing the CNS for damaged neurons and infectious agents. In this study, we compared microglia in the spinal cord of the young adult (1–2 years old) and aged (10–12 years old) German Shepherd dogs via immunohistochemistry and western blot analysis for ionized calcium-binding adapter molecule 1 (Iba-1), a microglial marker. In addition, we also observed the interferon-γ (IFN-γ), a pro-inflammatory cytokine, and interleukin-1β (IL-1β), produced by activated microglia/macrophage, protein levels in these groups. At first, we found that neuronal nuclei (NeuN, a neuronal marker)-immunoreactive neurons were distributed throughout the grey mate of the spinal cord, and there were no significant differences between the adult and aged groups. Most of Iba-1-immunoreactive microglia were morphologically ramified microglia (resting form) in the adult group, while some Iba-1-immunoreactive microglia were morphologically activated microglia in the aged group. In western blot analysis, Iba-1, IFN-γ and IL-1β expression were increased in the aged group. This result may be associated with age-dependent changes in the spinal cord.     

Comparison of Newly Generated Doublecortin-immunoreactive Neuronal Progenitors in the Main Olfactory Bulb among Variously Aged Gerbils

In the present study, we investigated age-related differences in neuronal progenitors in the gerbil main olfactory bulb (MOB) using doublecortin (DCX), a marker for neuronal progenitors which differentiate into neurons in the brain. No difference in the number of neuronal nuclei (NeuN)-immunoreactive neurons was found in the MOB at variously aged gerbils. At postnatal month (PM) 1, DCX immunoreaction was detected in all layers of the MOB except for the olfactory nerve layer. At this time point, DCX-immunoreactive cells (neuronal progenitors) were very abundant; however, they did not have fully developed-processes. From PM 3, the number of DCX-immunoreactive neuronal progenitors was decreased with age. At PM 6, DCX-immunoreactive cells showed very well-developed processes. In western blot analysis, DCX protein level in the MOB was highest at PM 1. Thereafter, levels of DCX protein were decreased with age. In the subventricular zone of the lateral ventricle, the number of Ki-67-immunoractive cells (proliferating cells) was also significantly decreased with age. In addition, increases of α-synuclein-immunoreactive structures were observed in the MOB with age. These results suggest that decrease in DCX-immunoreactive neuronal progenitors and its protein levels in the MOB with age may be associated with reduction of cell proliferation in the SVZ and with an increase in α-synuclein in the MOB.     

Cyclosporine A Reduces Dendritic Outgrowth of Neuroblasts in the Subgranular Zone of the Dentate Gyrus in C57BL/6 Mice

In the present study, we observed the effects of cyclosporine A (CsA), an efficient immunosuppressant, on cell proliferation and neuroblast differentiation in the subgranular zone of the dentate gyrus (SZDG) in normal C57BL/6 mice using Ki67 and doublecortin (DCX) immunohistochemical staining, respectively. At 8 weeks of age, vehicle (physiological saline) or CsA was daily administered (40 mg/kg, i.p.) for 1 week. Animals were sacrificed at 2 weeks after last administration. CsA treatment did not show any influences in neurons, astrocytes and microglia based on immunohistochemistry for its markers, respectively. However, in the CsA-treated group, Fluoro-Jade B, a marker for neurodegeneration, positive cells were found in the SZDG, not in the vehicle-treated group. In the vehicle-treated group, Ki67 immunoreactive (⁺) nuclei were clustered in the SZDG, whereas in the CsA-treated group Ki67⁺ nuclei were scattered in the SZDG, showing no difference in cell numbers. Numbers of DCX⁺ neuroblasts with well-developed processes (tertiary dendrites) were much lower in the CsA-treated group than those in the vehicle-treated group; however, numbers of DCX⁺ neuroblasts with secondary dendrites were similar in both the groups. These results suggest that CsA significantly reduces dendritic outgrowth and complexity from neuroblasts in the SZDG without any affecting in neurons, astrocytes and microglia in normal mice.     

EphA2 Mutation in Lung Squamous Cell Carcinoma Promotes Increased Cell Survival,Cell Invasion,Focal Adhesions,and Mammalian Target of Rapamycin Activation

Non-small cell lung cancer (NSCLC) has a poor prognosis and improved therapies are needed. Expression of EphA2 is increased in NSCLC metastases. In this study, we investigated EphA2 mutations in NSCLC and examined molecular pathways involved in NSCLC. Tumor and cell line DNA was sequenced. One EphA2 mutation was modeled by expression in BEAS2B cells, and functional and biochemical studies were conducted. A G391R mutation was detected in H2170 and 2/28 squamous cell carcinoma patient samples. EphA2 G391R caused constitutive activation of EphA2 with increased phosphorylation of Src, cortactin, and p130^Cas. Wild-type (WT) and G391R cells had 20 and 40% increased invasiveness; this was attenuated with knockdown of Src, cortactin, or p130^Cas. WT and G391R cells demonstrated a 70% increase in focal adhesion area. Mammalian target of rapamycin (mTOR) phosphorylation was increased in G391R cells with increased survival (55%) compared with WT (30%) and had increased sensitivity to rapamycin. A recurrent EphA2 mutation is present in lung squamous cell carcinoma and increases tumor invasion and survival through activation of focal adhesions and actin cytoskeletal regulatory proteins as well as mTOR. Further study of EphA2 as a therapeutic target is warranted.     

Carrier-mediated delivery of peptidic drugs for cancer therapy

Protein and peptide drugs are used for treatment of a variety of ailments. However, their wider use has been hindered by issues such as poor bioavailability in vivo and the cost involved in producing these drugs. This review discusses the various carrier-mediated methods used for delivery of peptide and protein drugs, with emphasis on liposomal and microspherical drug delivery systems. A brief look at the types of peptidic drugs currently in use clinically, and a brief discourse on several novel ideas for better protein delivery systems for cancer therapy is included.     

Paenibacillus hemolyticus, the first hemolytic Paenibacillus with growth-promoting activities discovered

Paenibacillus spp. are Gram-positive, facultatively aerobic, bacilli-shaped endospore-forming bacteria. They have been detected in a variety of environments, such as soil, water, forage, insect larvae, and even clinical samples. The strain 139SI (GenBank accession No.: JF825470.1) from three strains of Paenibacillus isolates investigated here was chosen as the type strain of the proposed novel species. The other two similar strain isolates investigated were 140SI (JF825471.1) and 141SI (JQ734548.1). These strains were identified as members of the genus Paenibacillus on the basis of phenotypic characteristics, phylogenetic analysis and 16S rRNA G+C content. Surprisingly, these strains exhibited a strong hemolytic activity on 5% sheep blood agar. Their crude extracts also showed positive growth-promoting activities in colon cancer and Vero cell lines. To our knowledge, this is the first Paenibacillus with hemolytic and growth-promoting activities reported, and the name Paenibacillus hemolyticus for this novel species is proposed. The capability of this novel species in hemolytic and cell growth activities suggests its potential in both clinical and pharmacological implications.     

Replacement of microglial cells using Clodronate liposome and bone marrow transplantation in the central nervous system of SOD1(G93A) transgenic mice as an in vivo model of amyotrophic lateral sclerosis

Disease progression of amyotrophic lateral sclerosis (ALS) is partially mediated by the toxic microenvironment established by microglia. In the present study, we used SOD1G93A transgenic mice as an in vivo ALS model and replaced microglia expressing mutant SOD1 (mSOD1) with microglia expressing wild-type SOD1 (w/tSOD1) to modulate the toxic microenvironment. Stereotactic injection of Clodronate liposome, a selective toxin against the monocyte/macrophage system, into the fourth ventricle of the brains of 12-week-old asymptomatic ALS mice reduced the number of microglia effectively in the central nervous system. Subsequent bone marrow transplantation (BMT) with bone marrow cells (BMCs) expressing w/tSOD1 and GFP leads to replacement of the endogenous microglia of the ALS mice with microglia expressing w/tSOD1 and GFP. The expression of mSOD1 in the other neural cells was not influenced by the replacement procedures, and immunological side effects were not observed. The replacement of microglia significantly slowed disease progression and prolonged survival of the ALS mice compared with the ALS mice treated by stereotactic injection of PBS-liposome and BMT with BMCs expressing mSOD1 or w/tSOD1. These results suggest that replacement of microglia would improve the neural cell microenvironment, thereby slowing disease progression. The mechanisms and functional implications of this replacement require further elucidation.