The present invention relates to the field of hyper-blebbing Gram-negative bacterial cells which are genetically modified by modifying the rpsA gene, the rpsA operon and/or 30S ribosomal protein S1 and to native outer membrane vesicles (nOMVs) obtained or obtainable from said genetically modified bacterial cells.

The present invention describes a second-generation tetanus toxoid vaccine and a process for the preparation thereof, comprising the steps of: inducing an E. Coli culture OD 600=0.5 by adding 0.2 mM IPTG; growing the culture at 14-16? C. for 14 to 20 hours; suspending the culture in 25 mM phosphate buffer containing 200 mM sodium chloride; adding 1% of triton-X-100 to the phosphate buffer, and adding the buffer to the culture; sonicating the culture for a period of 3 minutes (at 5 sec on/off pulse) at 4? C. on cold beads; centrifuging the culture for 60 to 90 minutes; collecting and purifying a supernatant using Ni-NTA affinity column with an eluant; and combining the supernatant into a pool with contaminated bands and concentrating using Centriprep-30 centrifuge filters (30 kDa pores).

The present invention describes a second-generation tetanus toxoid vaccine and a process for the preparation thereof, comprising the steps of: inducing an E. Coli culture OD 600=0.5 by adding 0.2 mM IPTG; growing the culture at 14-16? C. for 14 to 20 hours; suspending the culture in 25 mM phosphate buffer containing 200 mM sodium chloride; adding 1% of triton-X-100 to the phosphate buffer, and adding the buffer to the culture; sonicating the culture for a period of 3 minutes (at 5 sec on/off pulse) at 4? C. on cold beads; centrifuging the culture for 60 to 90 minutes; collecting and purifying a supernatant using Ni-NTA affinity column with an eluant; and combining the supernatant into a pool with contaminated bands and concentrating using Centriprep-30 centrifuge filters (30 kDa pores).

The present disclosure provides a kit and method for constructing a long-acting depression animal model. The present disclosure adopts Bacille Calmette-Guerin (BCG) (or Mycobacterium bovis (M bovis)) and low-dose pertussis toxin (PTX) for combined induction, which greatly reduces modeling cost. A mouse model constructed by the method of the present disclosure can clearly distinguish a time interval of pathological behaviors from a time interval of depression-like behaviors and has prominent face validity and predictive validity. The method of the present disclosure is simple and convenient and does not require a large number of tedious daily operations as the depression model construction by chronic unpredictable stress, chronic restraint stress, social defeat, mother-child separation, or the like. The mouse model of the present disclosure can provide a valuable long-term intervention window for research on antidepressant therapy.

The present disclosure provides a kit and method for constructing a long-acting depression animal model. The present disclosure adopts Bacille Calmette-Guerin (BCG) (or Mycobacterium bovis (M bovis)) and low-dose pertussis toxin (PTX) for combined induction, which greatly reduces modeling cost. A mouse model constructed by the method of the present disclosure can clearly distinguish a time interval of pathological behaviors from a time interval of depression-like behaviors and has prominent face validity and predictive validity. The method of the present disclosure is simple and convenient and does not require a large number of tedious daily operations as the depression model construction by chronic unpredictable stress, chronic restraint stress, social defeat, mother-child separation, or the like. The mouse model of the present disclosure can provide a valuable long-term intervention window for research on antidepressant therapy.

A mutated Bordetella strain comprising at least a mutated ptr gene, a deleted or mutated dnt gene and a heterologous ampG gene is provided. The attenuated mutated Bordetella strain can be used in an immunogenic composition or a vaccine for the treatment or prevention of a Bordetella infection. Use of the attenuated Bordetella strain for the manufacture of a vaccine or immunogenic composition, as well as methods for protecting mammals against infection by Bordetella are also provided.

A mutated Bordetella strain comprising at least a mutated ptx gene, a deleted or mutated dnt gene and a heterologous ampG gene is provided. The attenuated mutated Bordetella strain can be used in an immunogenic composition or a vaccine for the treatment or prevention of a Bordetella infection. Use of the attenuated Bordetella strain for the manufacture of a vaccine or immunogenic composition, as well as methods for protecting mammals against infection by Bordetella are also provided.

The present invention discloses methods and materials for delivering a cargo compound into a cancer cell. Delivery of the cargo compound is accomplished by the use of protein transduction domains derived from cupredoxins. The invention further discloses methods for treating cancer and diagnosing cancer.

The present invention discloses methods and materials for delivering a cargo compound into a cancer cell. Delivery of the cargo compound is accomplished by the use of protein transduction domains derived from cupredoxins. The invention further discloses methods for treating cancer and diagnosing cancer.

The present invention describes a second-generation tetanus toxoid vaccine and a process for the preparation thereof, comprising the steps of: inducing an E. Coli culture OD 600=0.5 by adding 0.2 mM IPTG; growing the culture at 14-16? C. for 14 to 20 hours; suspending the culture in 25 mM phosphate buffer containing 200 mM sodium chloride; adding 1% of triton-X-100 to the phosphate buffer, and adding the buffer to the culture; sonicating the culture for a period of 3 minutes (at 5 sec on/off pulse) at 4? C. on cold beads; centrifuging the culture for 60 to 90 minutes; collecting and purifying a supernatant using Ni-NTA affinity column with an eluant; and combining the supernatant into a pool with contaminated bands and concentrating using Centriprep-30 centrifuge filters (30 kDa pores).