Lactoferrin/pectin nanocomplex encapsulating ciprofloxacin and naringin as a lung targeting antibacterial nanoplatform with oxidative stress alleviating effect.

Pseudomonas aeruginosa is an opportunistic Gram-negative bacterium with adaptive metabolic abilities. It can cause hospital-acquired infections with significant mortality rates, particularly in people with already existing medical conditions. Its ability to develop resistance to common antibiotics makes managing this type of infections very challenging. Furthermore, oxidative stress is a common consequence of bacterial infection and antibiotic therapy, due to formation of reactive oxygen species (ROS) during their mode of action. In this study we aimed to alleviate oxidative stress and enhance the antibacterial efficacy of ciprofloxacin (CPR) antibiotic by its co-encapsulation with naringin (NAR) within a polyelectrolyte complex (PEX). The PEX comprised of polycationic lactoferrin (LF) and polyanionic pectin (PEC). CPR/NAR-loaded PEX exhibited spherical shape with particle size of 237 ± 3.5 nm, negatively charged zeta potential (-23 ± 2.2 mV) and EE% of 61.2 ± 4.9 for CPR and 76.2 ± 3.4 % for NAR. The LF/PEC complex showed prolonged sequential release profile of CPR to limit bacterial expansion, followed by slow liberation of NAR, which mitigates excess ROS produced by CPR's mechanism of action without affecting its efficacy. Interestingly, this PEX demonstrated good hemocompatibility with no significant in vivo toxicity regarding hepatic and renal functions. In addition, infected mice administrated this nanoplatform intravenously exhibited significant CFU reduction in the lungs and kidneys, along with reduced immunoreactivity against myeloperoxidase. Moreover, this PEX was found to reduce the lungs´ oxidative stress via increasing both glutathione (GSH) and catalase (CAT) levels while lowering malondialdehyde (MDA). In conclusion, CPR/NAR-loaded PEX can offer a promising targeted lung delivery strategy while enhancing the therapeutic outcomes of CPR with reduced oxidative stress.

Modulation of autophagy and apoptosis can contribute to the anticancer effect of Abemaciclib/Celecoxib combination in colon cancer cells.

Drug resistance and recurrence represent a great challenge in colorectal cancer management, highlighting the urgent need for novel therapeutics. Our objective is to evaluate the influence of Abemaciclib, Celecoxib, and their combination on both the autophagic and apoptotic machinery in an attempt to unravel the interplay between them in HCT-116 and Caco-2 cell lines. The MTT assay was used to assess the GI50 of the drugs. ELIZA was used to determine the protein levels of Beclin-1, LC3, Cox-2, and Bcl-2. Active Caspase-3 was determined by a colorimetric assay. Gene expression levels of ATG5, LC3, Beclin-1, and p62 were assessed by quantitative real-time PCR. In HCT-116 cells, the GI50s for Abemaciclib and Celecoxib were 15.86 and 92.67 µM, respectively, while for Caco-2 cells, the GI50s were 7.85 and 49.02 µM for Abemaciclib and Celecoxib, respectively. Upon treatment of HCT-116 and Caco-2 cells with Abemaciclib, Celecoxib, and their combinations, ATG5, p62, LC3, and Beclin-1 gene expression levels were up-regulated. The protein levels of Beclin-1, LC3, and Caspase-3 were significantly increased, while Bcl-2 was decreased in both cell lines due to single and combined treatments. Both drugs, either alone or in combination, decreased the migration ability of the cells in both cell lines. To conclude, the treatment protocol has the potential to induce cell cycle arrest, diminish the potentiality of cells for migration, and initiate apoptotic and autophagic cell death. Further research is recommended to unravel the potential antitumor effects of Abemaciclib/Celecoxib combination in different cancer types.

Electrospun polyvinyl alcohol/Withania somnifera extract nanofibers incorporating tadalafil-loaded nanoparticles for diabetic ulcers.

Background: Impaired inflammation and vascularization are common reasons for delayed diabetic wound healing. Nanoparticles (NPs)-in-nanofibers composites can manage diabetic wounds. A multifunctional scaffold was developed based on tadalafil (TDF)-loaded NPs incorporated into polyvinyl alcohol/Withania somnifera extract nanofibers. Materials & methods: TDF-loaded NPs were prepared and fully characterized in terms of their physicochemical properties. Extract of ashwagandha was prepared and a blend composed of TDF-loaded NPs, herbal extract and polyvinyl alcohol was used to prepare the whole composite. Results: The whole composite exhibited improved wound closure in a diabetic rat model in terms of reduced inflammation and enhanced angiogenesis. Conclusion: Results suggest that this multifunctional composite could serve as a promising diabetic wound dressing.

Chitosan Nanoparticles Alleviated the Adverse Effects of Sildenafil on the Oxidative Stress Markers and Antioxidant Enzyme Activities in Rats.

Sildenafil (SF) is widely used for erectile dysfunction and other conditions, though with limitations regarding oral absorption and adverse effects. Despite nanotechnological improvements, the effect of nanocarriers on SF hepatotoxicity has not been documented to date. This study aimed at assessing the impact of chitosan nanoparticles either uncoated (CS NPs) or Tween 80-coated (T-CS NPs) on the effects of SF on oxidative stress markers and antioxidant enzyme activities in rats. Test SF-CS NPs prepared by ionic gelation were uniform positively charged nanospheres (diameter 178-215 nm). SF was administered intraperitoneally to male rats (1.5 mg/kg body weight) in free or nanoencapsulated forms as SF-CS NPs and T-SF-CS NPs for 3 weeks. Free SF significantly suppressed the activity of the antioxidant enzymes glutathione S-transferase (GST), glutathione peroxidase (GPx), glutathione reductase (GR), catalase (CAT), and superoxide dismutase (SOD), as well as the levels of glutathione (GSH) and thiobarbituric acid reactive substances (TBARS) as in an indirect measure of free radicals. Interestingly, SF-CS NPs and T-SF-CS-NPs treatments significantly attenuated the inhibitory effects of SF on the activity of these enzymes whereas, GST activity was inhibited. Moreover, the protein expression of GST was downregulated upon treatment of rats with free SF, SF-CS-NPs, and T-SF CS-NPs. In contrast, the activity and protein expression of GPx was induced by SF-CS NPs and T-SF-CS-NPs treatments. The histopathological study showed that SF induced multiple adverse effects on the rat liver architecture which were markedly suppressed particularly by T-SF-CS NPs. In conclusion, chitosan nanoencapsulation of SF counteracted the adverse effects of SF on the activity of antioxidant enzymes and liver architecture. Findings might have significant implications in improving the safety and efficacy of SF treatment of the widely expanding disease conditions.

Naringin-loaded Arabic gum/pectin hydrogel as a potential wound healing material.

Wound healing is a complicated cellular process with overlapping phases. Naringin (NAR); a flavanone glycoside, possesses numerous pharmacological effects such as anti-inflammatory, antioxidant and anti-apoptotic effects. In the current study, Arabic gum (AG)/pectin hydrogel was utilized to encapsulate NAR. Drug-loaded AG/pectin hydrogel exhibited excellent EE% of about 99.88 ± 0.096 and high DL% of about 16.64 ± 0.013. The formulated drug-loaded hydrogel was characterized using Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), scanning electron microscopy (SEM) and Zetasizer analyzer, besides determination of equilibrium degree of swelling (EDS%). Afterwards, wound healing potential of NAR-loaded AG/pectin hydrogel was evaluated in an in vivo animal model. Results manifested that NAR-loaded AG/pectin hydrogel was able to accelerate wound healing in terms of enhanced angiogenesis, re-epithelialization and collagen deposition. Furthermore, it significantly (P < 0.001) down-regulated the mRNA expression of inflammatory mediators (TNF-α) and apoptosis (BAX). In addition, NAR-loaded AG/pectin hydrogel was found to possess potent antioxidant activity as it enhanced the levels of SOD and GSH, besides decreasing the levels of MPO, MDA and nitrite. These data suggest that NAR-loaded AG/pectin hydrogel could be utilized in wound healing applications.

GANT61/BI-847325 combination: a new hope in lung cancer treatment.

Despite the huge efforts employed to implement novel chemotherapeutic paradigms for lung cancer, the disease still remains a major concern worldwide. Targeting molecular pathways as Hedgehog (Hh) and Mitogen-activated protein kinase (MAPK) represent a new hope in lung cancer treatment. This work was undertaken to evaluate the antitumor effects of GANT61 (5 µM), BI-847325(30 µM), and GANT61 (5 µM)/BI-847325(30 µM) combination on A549 adenocarcinoma lung cancer cell line. The growth inhibition 50 (GI50) for both drugs was performed using MTT. The protein levels of Caspase-3, Bcl-2-associated X protein (Bax), Myeloid cell leukemia sequence 1 (MCL-1), cyclin D1, vascular endothelial growth factor (VEGF), extracellular signal-regulated kinases (ERK), p-Akt, and phosphohistone H3 (pHH3) were measured using ELISA. Glioma-associated oncogene homolog 1(Gli1) gene expression was assessed by quantitative real-time PCR. The GI50 for GANT61 and BI-8473255 were 5 µM and 30 µM, respectively. Caspase-3 and Bax protein levels were significantly elevated while MCL-1, cyclin D1, VEGF, ERK 1/2, p-Akt, and pHH3 levels were significantly reduced by both drugs and their combination relative to the control group. Gli1 gene expression was down-regulated in all groups relative to the control group. GANT61, BI-847325 and their combination inhibited proliferation and angiogenesis but activated the apoptotic pathway. Both drugs conferred a profound negative impact on the crosstalk between each of Hh and MAPK pathways and Phosphoinositide 3 -kinases (PI3K)/Akt/Mammalian target of Rapamycin (mTOR). To the best of our knowledge, the antitumor effects of BI-847325/GANT61 combination have not been tested before. Further in-vitro and in-vivo studies are warranted to support the findings.

Unraveling the therapeutic potential of GANT61/Dactolisib combination as a novel prostate cancer modality.

Aberrant activation of several signaling pathways has been implicated in prostate cancer (PCa) progression to castrate-resistant prostate cancer (CRPC). Phosphoinositide-3-kinase/Protein Kinase B/mechanistic Target of Rapamycin (PI3K/AKT/mTOR) and Hedgehog/GLI (Hh/GLI) pathways are major participants in progression to CRPC. In this sense, the current work aims to assess the potential antitumor effects resulting from co-targeting the aforementioned pathways in PC3 cells with Dactolisib as a dual PI3K/mTOR inhibitor and GANT61 as a GLI1 antagonist. Three replica of PC3 cells were assigned for four treatment groups; vehicle control, Dactolisib-treated, GANT61-treated, and combination-treated groups. GLI1 gene expression was determined by quantitative real-time PCR while active caspase-3 was determined colorimetrically. P-AKT, p70 ribosomal s6 protein kinase 1 (pS6K1), cyclin D1, vascular endothelial growth factor 1 (VEGF1), and Microtubule-associated proteins 1A/1B light chain 3 (LC3) protein levels were determined by ELISA technique. GLI1 gene expression was down-regulated as a result of Dactolisib, GANT61, and their combination. Additionally, both drugs significantly reduced p-AKT, pS6K1, cyclin D1, and VEGF1 protein levels. Dactolisib elevated LC3 protein levels and GANT61 augmented Dactolisib effect on LC3. Moreover, only Dactolisib/GANT61combination significantly increased active caspase-3 level. To sum up, Dactolisib/GANT61 combination was shown to be promising in PCa treatment. Further in-vitro and in-vivo studies are warranted to support our findings.

The biopolymer ulvan from Ulva fasciata: Extraction towards nanofibers fabrication.

As a biocompatible polymer, ulvan has applications in countless fields. Therefore, the following study intends to extract ulvan from Ulva fasciata, emphasizing its use for biomedical and industrial applications in the manufacture of nanofibrous webs. The extracted ulvan was characterized using FT-IR, DSC, XRD, GPC, and NMR. The extracted ulvan's FT-IR spectra confirmed that it is a sulfated polysaccharide. The HPLC analysis showed that the extracted ulvan is composed of rhamnose, xylose, glucose and glucuronic acid. NMR showed that the proton chemical shifts at 1.3 are due to methyl protons of rhamnose 3-sulfate in the ulvan samples. The X-ray diffractograms suggested that the extracted ulvan is semi-crystalline polymer with major crystalline reflection at 2θ of 29.4°. Deionized water has been successfully used to produce ulvan/polyvinyl alcohol (ulvan/PVA) nanofibers as an eco-friendly solvent. It was found that the ulvan-to-PVA (1:2) ratio results in nanofiber that is well handled and smooth. In addition to pretreated ones, the ulvan extracted without organic solvent pretreatment showed bead free nanofibers. It is concluded that pretreatment with organic solvent in ulvan extraction, particularly in the manufacture of nanofibers, is not recommended. In addition, the resulting nanofibrous mat has sufficient mechanical properties for various applications to be incorporated.

Dual targeting of Notch and Wnt/ß-catenin pathways: Potential approach in triple-negative breast cancer treatment.

Despite the continuously growing repertoire of new and improved anti-cancer therapies, triple-negative breast cancer (TNBC) remains a clinical challenge to treat. In this sense, targeting signaling pathways such as Notch and Wnt/ß-catenin have attracted growing attention. This work aimed at investigating the possible antitumor effects of IMR-1 as a Notch inhibitor, PRI-724 as a Wnt/ß-catenin inhibitor, as well as their combination and to explore the possible crosstalk between Notch and Wnt/ß-catenin signaling pathways in MDA-MB-231 TNBC cell line. Microculture tetrazolium test (MTT) was used to determine the drug growth inhibition (GI50), and the results were analyzed using CompuSyn 3.0.1 software. MDA-MB-231 cells were divided into four treatment groups including positive control, IMR-1-treated, PRI-724-treated, and combination-treated groups. Sandwich enzyme-linked immunosorbent assay (ELISA) was used for the determination of the protein levels of hairy and enhancer of split-1 (HES-1), Notch-1, ß-catenin, cyclin-D1, and vascular endothelial growth factor (VEGF1). HES-1 gene expression was assessed by quantitative real-time polymerase chain reaction. Statistical analyses were performed using GraphPad Prism Software. The GI50 for IMR-1 and PRI-724 were 15.3 µM and 0.69 µM, respectively. Upon treatment of MDA-MB-231 cells with these drugs, HES-1 gene expression was up-regulated due to single and combined treatments. Moreover, the protein levels of cyclin-D1, VEGF1, HES-1, and Notch-1 were reduced, while those of active ß-catenin and active caspase-3 were elevated. IMR-1/PRI-724 combination augmented IMR-1- and PRI-724-mediated effects on MDA-MB-231 cells by initiating apoptotic cell death. Further in vitro and in vivo studies are warranted to support our findings.

Association of matrix metalloproteinases 3 and 9 single nucleotide polymorphisms with breast cancer risk: A case-control study.

Two single nucleotide polymorphisms (SNPs) of matrix metalloproteinase (MMPs) 3 and 9 are functionally implicated in the progression of various types of cancer, including breast cancer (BC). However, the roles of these SNPs remain controversial. In addition, they also vary between one population and another. Therefore, the present study aimed to investigate the possible association between MMP3-1171 5A/6A and MMP9-1562 CT SNPs and the risk of BC among Egyptians, and to elucidate the alteration of MMP3 and MMP9 gene expression in patients with BC. The present case-control study enrolled 162 patients with BC and 146 control subjects. Restriction fragment length polymorphism-PCR was performed for analysis of the selected SNPs, gene expression of MMP3 and MMP9 was also assessed in 50 patients and 50 control subjects by reverse transcription-quantitative PCR. The frequencies of 5A/6A genotype and 5A allele of MMP3 were significantly higher in patients with BC compared with in healthy subjects. On the other hand, the distributions of MMP9 genotypes and alleles were not significantly different among patients and healthy subjects. Compared with healthy subjects, the expression levels of the two genes were found to be upregulated in patients with BC. Therefore, the present study indicated that MMP3-1171 5A/6A SNP, not MMP9-1562 C>T SNP may be a risk factor for developing BC among Egyptian females.

Recombinant N-terminal outer membrane porin (OprF) of Pseudomonas aeruginosa is a promising vaccine candidate against both P. aeruginosa and some strains of Acinetobacter baumannii.

Pseudomonas aeruginosa is an evolving pathogen which can cause serious infections especially to immunocompromised patients. Its high resistance profile to antibiotics results in difficulty, and sometimes impossibility, in treating afflicted patients. Developing an effective vaccine against P. aeruginosa is an important approach to tackle this problem. A similar problematic situation exists for Acinetobacter baumannii. Several vaccine candidates have been investigated up till now but still there is no approved vaccine in the market. One important antigen of P. aeruginosa is the outer membrane protein F (OprF) which functions as a porin with relevant important roles in virulence. Previous studies focused mainly on the C-terminal peptidoglycan binding domain of OprF as a vaccine candidate. In the current study, we have investigated the N-terminal porin domain of OprF as a potential vaccine candidate against P. aeruginosa. Histidine-tagged recombinant N-terminal OprF (amino acid range 25-200; OprF25-200) was overexpressed in Escherichia coli and purified using metal affinity chromatography. Swiss albino mice were immunized with OprF25-200 adjuvanted with Bacillus Calmette-Guérin (BCG) and alum and the immune response was evaluated. Immunized mice developed antigen-specific IgG1 and IgG2a and were protected against challenge by both P. aeruginosa and a clinical isolate of A. baumannii expressing OprF. Serum from OprF25-200-immunized mice showed cross-reactivity with both pathogens using western blotting and whole cell enzyme-linked immunosorbent assay (ELISA). To our knowledge, this is the first report to demonstrate that the N-terminal domain of OprF is sufficiently immunogenic to protect against the two pathogens.

Magnetically Guided Self-Assembled Protein Micelles for Enhanced Delivery of Dasatinib to Human Triple-Negative Breast Cancer Cells.

Magnetic nanocarriers are useful in targeted cancer therapy. Dasatinib (DAS)-loaded magnetic micelles were prepared for magnetically guided drug delivery. The magnetic nanoplatform is composed of hydrophobic oleic acid-coated magnetite (Fe3O4) core along with DAS encapsulated in amphiphilic zein-lactoferrin self-assembled polymeric micelles. Transmission electron microscope analysis manifested formation of these magnetic micelles with a mean diameter of about 100 nm. In addition, drug-loaded magnetic micelles displayed a saturation magnetization of about 10.01 emu.g-1 with a superparamagnetic property. They also showed good in vitro serum stability and hemocompatibility accompanied with a sustained release of DAS in acidic pH. More importantly, they exhibited 1.35-fold increase in their in vitro cytotoxicity against triple-negative human breast cancer cell line (MDA-MB-231) using an external magnetic field compared to drug-loaded magnetic micelles in the absence of a magnetic field. Enhanced inhibition of p-c-Src protein expression level and in vitro cellular migration under the effect of magnetic field was noted owing to the dual-targeting strategy offered by the presence of a magnetic sensitive core, as well as the active targeting property of lactoferrin corona. Taken all together, these results suggest that DAS-loaded magnetic micelles possess a great potential for targeted therapy of breast cancer.

The Enhancement of the Pasteurella's Bacterin by Propolis Extracts.

BACKGROUND: Pasteurella multocida continues to pose a danger to prone farm and wild animals all over the world. Chemotherapeutic treatments are progressively losing their effectiveness, last for long time, and cost a lot of money, as well as being toxic to human consumers. Therefore, clearing the way for immunization as a big-wheel alternative against the economic grain. Yet, the vaccines available in the market do not confer the necessary protection against the pathogen. The integration of the well adjuvanted killed vaccine with the attenuated vaccines proved to offer an effective protection to the host animals. However, the bare use of the killed bacterin to provide protection from the possible harm of the live attenuated vaccine was doubtful. METHODS: In the present study, propolis extracts were used to ameliorate the immunogenicity of the Pasteurella bacterin. The cellular and humoral activities were assessed for the different bacterin formulations. RESULTS: Propolis extracts adjuvants proved to broaden and extend the IgG potency, as well as to induce a unique mucosal protection against the bacterium. Simultaneously it offered an anti-inflammatory effect that increased the tolerability to the bacterin. While the cellular activity was relatively reduced with propolis extracts. CONCLUSION: These results confirm the effectiveness of the formulation of the bacterin with propolis to offer a potent homologous primary protection to the animals against the long-life use of the attenuated Pasteurella vaccines.

Self-assembled amphiphilic zein-lactoferrin micelles for tumor targeted co-delivery of rapamycin and wogonin to breast cancer.

Protein-based micelles have shown significant potential for tumor-targeted delivery of anti-cancer drugs. In this light, self-assembled nanocarriers based on GRAS (Generally recognized as safe) amphiphilic protein co-polymers were synthesized via carbodiimide coupling reaction. The new nano-platform is composed of the following key components: (i) hydrophobic zein core to encapsulate the hydrophobic drugs rapamycin (RAP) and wogonin (WOG) with high encapsulation efficiency, (ii) hydrophilic lactoferrin (Lf) corona to enhance the tumor targeting, and prolong systemic circulation of the nanocarriers, and (iii) glutaraldehyde (GLA)-crosslinking to reduce the particle size and improve micellar stability. Zein-Lf micelles showed relatively rapid release of WOG followed by slower diffusion of RAP from zein core. This sequential release may aid in efflux pump inhibition by WOG thus sensitizing tumor cells to RAP action. Interestingly, these micelles showed good hemocompatibility as well as enhanced serum stability owing to the brush-like architecture of Lf shell. Moreover, this combined nano-delivery system maximized synergistic cytotoxicity of RAP and WOG in terms of tumor inhibition in MCF-7 breast cancer cells and Ehrlich ascites tumor animal model as a result of enhanced active targeting. Collectively, GLA-crosslinked zein-Lf micelles hold great promise for combined RAP/WOG delivery to breast cancer with reduced drug dose, minimized side effects and maximized anti-tumor efficacy.

The detection of antigenic determinants of Acinetobacter baumannii.

BACKGROUND: Acinetobacter baumannii continues to pose a threat to burdened patients in ICUs all around the world. Lately, infection control techniques are not sufficient to curb A. baumannii's progression and chemotherapeutics are losing their potency against it. Thus, immunization became a key player in providing an ideal solution to the dilemma. None of the vaccines under investigation have reached the market and the search for a tailored vaccine remains a challenge. The notion of unravelling the bacterial antigens to design a novel epitope-based vaccine proved its merits. METHODS: In this work, the propitious polysaccharide and protein antigenic determinants of A. baumannii were mapped by mimicking the infection. The immune response was evaluated by western blot, ELISA, and cellular proliferation assay techniques. RESULTS: The screening showed that OMPs induced the most eminent sustained IgG response. In addition, OMP gave the highest cellular proliferation and a fold increase in ELISA that reached up to 10-fold by week 6. Whilst, the LPS gave a rapid IgM response, that reached 5-fold and the response was visible from week 1 in the western blot. The OMPs had a more pronounced effect in eliciting a cellular immune response. CONCLUSION: The results elaborated the valuable role of using pure OMPs and detoxified LPS together; as a major cornerstone in designing an ideal vaccine against A. baumannii.

Development of immunization trials against Pasteurella multocida.

Pasteurellosis is one of the most important respiratory diseases facing economically valuable farm animals such as poultry, rabbit, cattle, goats and pigs. It causes severe economic loss due to its symptoms that range from primary local infection to fatal septicemia. Pasteurella multocida is the responsible pathogen for this contagious disease. Chemotherapeutic treatment of Pasteurella is expensive, lengthy, and ineffective due to the increasing antibiotics resistance of the bacterium, as well as its toxicity to human consumers. Though, biosecurity measures played a role in diminishing the spread of the pathogen, the immunization methods were always the most potent preventive measures. Since the early 1950s, several trials for constructing and formulating effective vaccines were followed. This up-to-date review classifies and documents such trials. A section is devoted to discussing each group benefits and defects.

Development of a new trend conjugate vaccine for the prevention of Klebsiella pneumoniae.

Klebsiella pneumoniae is a major cause of nosocomial pneumonia, septicemia and urinary tract infections, especially in newborns, blood cancer patients, and other immunocompromised candidates. The control of K. pneumoniae is a complicated issue due to its tight pathogenesis. Immuno-prophylactic preparations, especially those directed toward the bacterium O-antigen, showed to be the most successful way to prevent the infection incidence. However, all previously proposed preparations were either of limited spectrum or non-maternal, and hence not targeting the main Klebsiella patients. Moreover, all preparations were directed only to prevent the respiratory diseases due to that pathogen. This article addresses the development of a method originally used to purify the non-capsular bacterial-endotoxins, as a new and easy method for vaccine production against K. pneumoniae. The application of this method was preceded by a biotechnological control of capsular polysaccharide production in K. pneumoniae. The new produced natural conjugate between the bacterial O-antigen and its outer membrane proteins was evaluated by physicochemical and immunological methods to investigate its purity, integrity, safety and immunogenicity. It showed to be pure, stable, safe for use, and able to elicit a protective immunoglobulin titer against different Klebsiella infections. This immune-response proved to be transferable to the offspring of the vaccinated experimental rabbits via placenta.

Development of immunization trials against Klebsiella pneumoniae.

Klebsiella pneumoniae is the most common cause of nosocomial respiratory tract and premature intensive care infections, and the second most frequent cause of Gram-negative bacteraemia and urinary tract infections. Drug resistant isolates remain an important hospital-acquired bacterial pathogen, add significantly to hospital stays, and are especially problematic in high impact medical areas such as intensive care units. Many investigations worldwide proved the increasing resistance of such pathogen, resulting in an average rate of 1.63 outbreak every year. A variety of preventive measures were applied to reduce such incidences. Immunotherapy and passive immunization researches as well found their way to the treatment of Klebsiella. During the last 40 years, many trials for constructing effective vaccines were followed. This up-to-date review classifies such trials and documents them in a progressive way. A following comment discusses each group benefits and defects.

Measurement of IgG levels can serve as a biomarker in newly diagnosed diabetic children.

This study was undertaken to determine humoral immune response to the presence of anti-immunoglobulin antibodies in children with newly diagnosed type 1 diabetes mellitus, using as a target cow immunoglobulins, in an attempt to elucidate further complex immuno-pathogenetic interactions of the disease. Serum immunoglobulin G (IgG) concentrations were measured by ELISA in 30 children with type 1 diabetes mellitus and 30 healthy matched normal children. It was found that normal children had a mean IgG level of 7.41 mg/ml while diabetic individuals had a mean IgG level of 8.52 mg/ml (p<0.00004). On the contrary, the mean level of IgG in diabetic sera after purification from anti-cow immunoglobulins was determined to be 7.52 mg/ml. Therefore, there was no significant difference in IgG level in patients with type 1 diabetes mellitus after removal of anti-cow immunoglobulin antibodies compared to normal children (p<0.58). Visualization of IgG and immuno-precipitation confirm that anti-cow immunoglobulins antibodies, which were unrelated to antigen, were co-precipitated with the antigen-antibody complex. A circulating immunoglobulin reacting with other immunoglobulins is thus present in children with type 1 diabetes and may well play a part in the complex immuno-pathogenetic interactions.

The influence of tobacco smoking on humoral immune response in insulin dependent diabetic pregnancy.

Tobacco smoking products have a heavy impact on the public health of developed as well as non-developed countries by being a main etiologic factor for the induction of cardiovascular diseases and tobacco-related cancer. The purpose of this study was to determine the influence of tobacco smoking on the measurement of the humoral immune response in Egyptian pregnant women with type 1 diabetes. Concentrations of serum immunoglobulin A, G and M in 35 smoking, 35 non-smoking pregnant women with type 1 diabetes and 35 matched normal women were measured by ELISA. Women were matched by age and working life with controls. Measurements suggested that diabetic smokers had decreased levels of IgG and IgM in their sera. It was found that normal individuals had mean IgA, IgG and IgM levels of 2.80 mg/ml, 9.33 mg/ml and 1.66 mg/ml, respectively while non-smoker women suffering from type 1 diabetes had mean levels of 3.47 mg/ml, 10.97 mg/ml and 2.05 mg/ml (p<0.0004,p<0.0001 andp<0.0002). However, the mean level of IgA, IgG and IgM in diabetic smoker sera was determined to be 3.33 mg/ml, 8.07 mg/ml and 1.31 mg/ml, respectively (p<0.003,p<0.0001 andp<0.0001). The obtained results suggest that toxic smoke components were immuno-suppressant and may well play a part in the complex immuno-pathogenesis interaction. The increased risk of smoking in insulin dependent diabetic pregnant women during pregnancy is a further reason to encourage pregnant women to quit tobacco smoking.