Anti-mycobacterial recall responses differentiate female patients with genital tuberculosis from patients with other gynecological problems

Background: Female Genital Tuberculosis (FGTB) is one form of extra pulmonary tuberculosis affecting the female reproductive organs, most commonly the fallopian tubes and the endometrium. It affects young women aged between 20 and 40 years of age and is an important cause of infertility. It often occurs as a secondary complication following pulmonary tuberculosis. Diagnosis depends mainly on clinical suspicion in countries where facilities for mycobacterial culture and histopathology are unavailable. Even in places where these facilities exist, diagnosis still remains difficult because of the lower sensitivity and specificity of the methods as well as the invasive procedure of acquiring biopsy specimens. Objective: To explore the immunological profiles of female genital tuberculosis (FGTB) patients in response to mycobacterial antigens. Methods: Twenty-five clinically suspected cases of FGTB and 12 control subjects who came to the Black Lion hospital for unrelated gynecological problems were included in the study. Peripheral blood samples were collected from each subject. Plasma was separated by centrifugation and PBMC were isolated over ficoll-hypaque and stimulated in vitro with mycobacterial antigens to examine their proliferative response as incorporation of tritiated thymidine using a β-counter. HIV status and total IgG-, IgA- and IgM- antibody levels were determined by ELISA tests.Results: In vitro recall responses to M. tuberculosis antigens (PPD and BCG sonicate) as well as plasma levels of IgGIgA- and IgM-antibodies to MPT59 showed statistically significant differences between the patients and the controls (

Evaluation of a direct colorimetric assay for rapid detection of rifampicin resistant Mycobacterium tuberculosis

No Abstract Available Ethiop.J.Health Dev. Vol.19(1) 2005: 51-5

Novel approaches toward the development of an oral post-exposure DNA vaccine for latent tuberculosis using Salmonella typhimurium ΔaroA vector

Tuberculosis remains one of the major causes of global public health problems. There is no effective vaccine for the disease until now. Many reports show that DNA vaccines are promising to induce protection against Mycobacterium tuberculosis (M. tb); however, the efficiency of DNA vaccine is limited due to inadequate delivery systems. Among others, live attenuated bacterial vectors such as Salmonella enterica typhimurium (S. typhimurium) have significant promise as efficient mucosal delivery vehicles for DNA vaccines. In this study, we constructed recombinant attenuated S. typhimurium DNA vaccines carrying genes encoding resuscitation promoting factor (Rpf)-like proteins of M. tb on eukaryotic expression plasmid agianst latent tuberculosis and evaluated the plasmid stability and growth curve assays of the recombinant Salmonella vaccine constructs in vitro. Four Rpf gene fragments (RpfB, RpfC, RpfD, RpfE) associated with latency were amplified from genomic DNA of the H37Rv strain of M. tb, cloned into eukaryotic expression plasmid (pVR1020) and verified by sequencing. In later studies, we will demonstrate the potential use of the Salmonella-mediated DNA constructs as candidate post-exposure vaccines against tuberculosis through testing their immunogenicity and effectiveness for oral delivery in eukaryotic systems.Key words: Latent tuberculosis, resuscitation promoting factor (Rpf), DNA vaccine, recombinant Salmonella typhimurium

Encapsulation of Gadolinium Oxide Nanoparticle (Gd₂O₃) Contrasting Agents in PAMAM Dendrimer Templates for Enhanced Magnetic Resonance Imaging <i>in Vivo</i>

There has been growing interest in the research of nanomaterials for biomedical applications in recent decades. Herein, a simple approach to synthesize the G4.5-Gd₂O₃-poly­(ethylene glycol) (G4.5-Gd₂O₃-PEG) nanoparticles (NPs) that demonstrate potential as dual (<i>T</i>₁ and <i>T</i>₂) contrasting agents in magnetic resonance imaging (MRI) has been reported in this study. Compared to the clinically popular Gd-DTPA contrasting agents, G4.5-Gd₂O₃-PEG NPs exhibited a longer longitudinal relaxation time (<i>T</i>₁) and better biocompatibility when incubated with macrophage cell line RAW264.7 <i>in vitro</i>. Furthermore, the longitudinal relaxivity (<i>r</i>₁) of G4.5-Gd₂O₃-PEG NPs was 53.9 s^–1 mM^–1 at 7T, which is equivalent to 4.8 times greater than to the Gd-DTPA contrasting agents. An <i>in vivo</i> <i>T</i>₁-weighted MRI results revealed that G4.5-Gd₂O₃-PEG NPs significantly enhanced signals in the intestines, kidney, liver, bladder, and spleen. In addition, the <i>T</i>₂-weighted MRI results revealed darker contrast in the kidney, which proves that G4.5-Gd₂O₃-PEG NPs can be exploited as <i>T</i>₁ and <i>T</i>₂ contrasting agents. In summary, these findings suggest that the G4.5-Gd₂O₃-PEG NPs synthesized by an alternative approach can be used as dual MRI contrasting agents

IL‑6 Antibody and RGD Peptide Conjugated Poly(amidoamine) Dendrimer for Targeted Drug Delivery of HeLa Cells

In this study, PAMAM dendrimer (G4.5) was conjugated with two targeting moieties, IL-6 antibody and RGD peptide (G4.5-IL6 and G4.5-RGD conjugates). Doxorubicin anticancer drug was physically loaded onto G4.5-IL6 and G4.5-RGD with the encapsulation efficiency of 51.3 and 30.1% respectively. The cellular internalization and uptake efficiency of G4.5-IL6/DOX and G4.5-RGD/DOX complexes was observed and compared by confocal microscopy and flow cytometry using HeLa cells, respectively. The lower IC₅₀ value of G4.5-IL6/DOX in comparison to G4.5-RGD/DOX is indication that higher drug loading and faster drug release rate corresponded with greater cytotoxicity. The cytotoxic effect was further verified by increment in late apoptotic/necrotic cells due to delivery of drug through receptor-mediated endocytosis. On the basis of these results, G4.5-IL6 is a better suited carrier for targeted drug delivery of DOX to cervical cancer cells

Bioinspired, Manganese-Chelated Alginate–Polydopamine Nanomaterials for Efficient in Vivo <i>T</i>₁‑Weighted Magnetic Resonance Imaging

Manganese-based nanomaterials are an emerging new class of magnetic resonance imaging (MRI) contrast agents (CAs) that provide impressive contrast abilities. MRI CAs that can respond to pathophysiological parameters such as pH or redox potential are also highly in demand for MRI-guided tumor diagnosis. Until now, synthesizing nanomaterials with good biocompatibility, physiochemical stability, and good contrast effects remains a challenge. This study investigated two new systems of calcium/manganese cations complexed with either alginate–polydopamine or alginate–dopamine nanogels [AlgPDA­(Ca/Mn) NG or AlgDA­(Ca/Mn) NG]. Under such systems, Ca cations form ionic interactions via carboxylic acids of the Alg backbone to enhance the stability of the synthetic nanogels (NGs). Likewise, complexation of Mn cations also increased the colloidal stability of the synthetic NGs. The magnetic property of the prepared CAs was confirmed with superconducting quantum interference device measurements, proving the potential paramagnetic property. Hence, the <i>T</i>₁ relaxivity measurement showed that PDA-complexed synthetic NGs reveal a strong positive contrast enhancement with <i>r</i>₁ = 12.54 mM^–1·s^–1 in 7.0 T MRI images, whereas DA-complexed synthetic NGs showed a relatively lower <i>T</i>₁ relaxivity effect with <i>r</i>₁ = 10.13 mM^–1·s^–1. In addition, both the synthetic NGs exhibit negligible cytotoxicity with >92% cell viability up to 0.25 mM concentration, when incubated with the mouse macrophage (RAW 264.7) and HeLa cells, and high biocompatibility under in vivo analysis. The in vivo MRI test indicates that the synthetic NG exhibits a high signal-to-noise ratio for longer hours, which provides a longer image acquisition time for tumor and anatomical imaging. Furthermore, <i>T</i>₁-weighted MRI results revealed that PEGylated AlgPDA­(Ca/Mn) NGs significantly enhanced the signals from liver and tumor tissues. Therefore, owing to the enhanced permeability and retention effect, significantly enhanced in vitro and in vivo imagings, low cost, and one-pot synthesis method, the Mn-based biomimetic approach used in this study provides a promising and competitive alternative for noninvasive tumor detection and comprehensive anatomical diagnosis