Novel camel milk-derived proteins and their application in the management of acne vulgaris

Acne vulgaris, caused by the bacterium P. acnes, is the most common cutaneous disorder in adolescents. Acne is not a life-threatening condition, however, it has significant psychological and physical effects on patients. P. acnes is capable of causing remarkable inflammatory responses. Topical therapy is advisable in acne treatment in mild to moderate acne conditions. Recently, available treatments for acne are mostly based on antibiotics and retinoids. The use of antibiotics has the limitations of developing bacterial resistance, while retinoids are highly teratogenic. The bacterial resistance and teratogenicity of isotretinoin led scientists to search for more potent and safer therapeutic options. Camel milk has been used in the preparation of pharmaceutical and cosmetic compositions such as anti-acne products. It was found that camel milk composition is closer to the human milk. Passive immunity is provided to new-borns by Immunoglobulins present in colostrum until its own immune system matures. The concentration in colostrum of specific antibodies against pathogens can be raised by immunizing a mammal with these pathogens or their antigens. Immunized milk products are preparations made of such hyper-immune antibodies enriched from it. Camel milk and milk-derived products have a growing potential therapeutic value. The objective of this research is to evaluate the antimicrobial and anti-inflammatory effects of immunized camel milk whey and its components against P. acnes and to develop a formula that harnesses all its beneficial properties. P. acnes was heat-killed in culture to obtain their outer membrane proteins (OMPs). The OMPs were mapped and quantified using SDS-PAGE and Bradford Assay, respectively. OMPs were used to prepare a vaccination injected into female lactating camels in timely manner. The immunized camel milk was collected then pasteurized, delipidized, de-caseinated, and freeze-dried to obtain the whey into powder form. The anti-P.acnes antibodies in the immunized camel whey were separated using Protein G and Protein A Chromatography and quantified by SDS-PAGE and ELISA, respectively. Then, the antimicrobial peptides (AMPs) including PGRPs and Lactoferrin were isolated by Heparin Affinity Chromatography, and mapped by SDS-PAGE. The antimicrobial and anti-inflammatory activity of the immunized camel whey and its constituents against P. acnes was evaluated in vitro. In vivo activity was also evaluated using rabbit ear model. After the activity of the immunized camel whey was confirmed, four topical formulae were developed and their composition and preparation protocol were patented. The findings of this research imply that the constituents of immunized camel milk whey (i.e. polyclonal anti-P. acnes antibodies, peptidoglycan recognition proteins (PGRPs), and lactoferrin) possess significant antimicrobial and anti-inflammatory activity against P. acnes, 90% and 70%, respectively. Topical formulae including; facial wash, serum, cream and gel enriched with 3% to 5% of immunized camel milk whey were developed and their activity against P. acnes was evaluated. Immunized camel milk whey that has been developed in this research exerts significant antimicrobial and anti-inflammatory activity against P. acnes and the unique composition formulated has been patented. Furthermore, the stability of the topical cream was validated to show that the antibodies in the cream formula retained around 75% of their activity.Acne vulgaris, caused by the bacterium P. acnes, is the most common cutaneous disorder in adolescents. Acne is not a life-threatening condition, however, it has significant psychological and physical effects on patients. P. acnes is capable of causing remarkable inflammatory responses. Topical therapy is advisable in acne treatment in mild to moderate acne conditions. Recently, available treatments for acne are mostly based on antibiotics and retinoids. The use of antibiotics has the limitations of developing bacterial resistance, while retinoids are highly teratogenic. The bacterial resistance and teratogenicity of isotretinoin led scientists to search for more potent and safer therapeutic options. Camel milk has been used in the preparation of pharmaceutical and cosmetic compositions such as anti-acne products. It was found that camel milk composition is closer to the human milk. Passive immunity is provided to new-borns by Immunoglobulins present in colostrum until its own immune system matures. The concentration in colostrum of specific antibodies against pathogens can be raised by immunizing a mammal with these pathogens or their antigens. Immunized milk products are preparations made of such hyper-immune antibodies enriched from it. Camel milk and milk-derived products have a growing potential therapeutic value. The objective of this research is to evaluate the antimicrobial and anti-inflammatory effects of immunized camel milk whey and its components against P. acnes and to develop a formula that harnesses all its beneficial properties. P. acnes was heat-killed in culture to obtain their outer membrane proteins (OMPs). The OMPs were mapped and quantified using SDS-PAGE and Bradford Assay, respectively. OMPs were used to prepare a vaccination injected into female lactating camels in timely manner. The immunized camel milk was collected then pasteurized, delipidized, de-caseinated, and freeze-dried to obtain the whey into powder form. The anti-P.acnes antibodies in the immunized camel whey were separated using Protein G and Protein A Chromatography and quantified by SDS-PAGE and ELISA, respectively. Then, the antimicrobial peptides (AMPs) including PGRPs and Lactoferrin were isolated by Heparin Affinity Chromatography, and mapped by SDS-PAGE. The antimicrobial and anti-inflammatory activity of the immunized camel whey and its constituents against P. acnes was evaluated in vitro. In vivo activity was also evaluated using rabbit ear model. After the activity of the immunized camel whey was confirmed, four topical formulae were developed and their composition and preparation protocol were patented. The findings of this research imply that the constituents of immunized camel milk whey (i.e. polyclonal anti-P. acnes antibodies, peptidoglycan recognition proteins (PGRPs), and lactoferrin) possess significant antimicrobial and anti-inflammatory activity against P. acnes, 90% and 70%, respectively. Topical formulae including; facial wash, serum, cream and gel enriched with 3% to 5% of immunized camel milk whey were developed and their activity against P. acnes was evaluated. Immunized camel milk whey that has been developed in this research exerts significant antimicrobial and anti-inflammatory activity against P. acnes and the unique composition formulated has been patented. Furthermore, the stability of the topical cream was validated to show that the antibodies in the cream formula retained around 75% of their activity

In Vitro anti- Helicobacter pylori and urease inhibitory effects of polyphenolic extracts of local herbs from Algeria

Gastritis and peptic ulcers are considered as major health problems worldwide with more than 80% of chronic active gastritis are due to the pathogenic bacterium H. pylori. Due to the emergence of antibiotic resistance among clinical strains of H. pylori, alternative approaches are increasingly needed. Methanolic extracts of Algerian originated Mentha rotundifolia, Eucalyptus globulus, Malva sylvestris, Inula viscosa, Achillea odorata and Utrica dioica and their contents of phenolics and flavonoids were evaluated for the in vitro antimicrobial activity against H. pylori in addition to its associated urease. The antimicrobial activity and determination of the MICs of these extracts against control strain of H. pylori was performed using standard agar diffusion method.  The highest phenolic and flavonoid contents were found in M. rotundifolia and E. globulus while M. sylvestris showed the least phenolic contents. In addition, polyphenolic fractions exhibited anti-H. pylori activity of all of the herbal extracts with  highest activity for E. globulus (MIC 0.094 mg/ml), I. viscosa, (0.375 mg/ml) and U. dioica (0.75 mg/ml), low antimicrobial activity was revield for  A. odorata, M. rotundifolia and Malva sylvestris respectively (MIC > 1 mg/ml).  For the urease activity, all extracts showed inhibitory effect at concentration of 250 mg/ml. However, the range of the urease inhibitory concentrations varied significantly among the extracts with highest activity and widest range found for E. globulus (70-90% at concentrations 8-125 mg/ml). In conclusion, the presence of potential antimicrobial activities in the polyphenolic extracts of medicinal plants against H. pylori and its associated urease would open the door for investigation of novel agents for the control of H. pylori infections