Submit or Track your Manuscript LOG-IN

D.K. Hazra, Megha Pant, S.K. Raza and P. K. Patanjali

 

Formulation technology: key parameters for food safety with respect to agrochemicals use in crop protection
...n-degradable ingredients/adjuvants of the conventional formulations but also increase the bio-efficacy of the products through incorporating latest technologies including size reduction (WP to SC, SL to ME), increased coverage of applied surface area (EC to ME/Nano-formulations), reduced wastage (Dust/WP to Con-trolled Release Formulations) and dose rates of same pesticides applied to improve food quality with minimum pesticide residues. 

...
Mumtaz Ali Khan1, Aneela Zameer Durrani1, Sher Bahadar Khan2, Shehla Gul Bokhari1, Ikramul Haq1,*, Imdad Ullah Khan3, Naimat Ullah4, Naimat Ullah Khan4, Kashif Hussain1 and Azmat Ullah Khan2
...antified, formalized and adjuvanted with aluminium hydroxide gel. Safety, sterility and stability of the toxoid were ascertained. Bacterin toxoid of C. perfringens type D was procured from Veterinary Research Institute Peshawar. The immunogenicity of toxoid vaccines and bacterin toxoid vaccine were studied and compared initially in rabbits and then in goats and sheep. Experimental trials were conducted on 54 healthy animals of each species and were divi...
Yan-Guo Han1, Xiao-Li Peng1, Kai Li1, Yu-He-Tian Zhao1, Xun-Ping Jiang2, Guang-Xin E1, Yong-Ju Zhao1, Jun-Hua Ye3, Li Xu3, Qin-Tao Zhao3 and Yong-Fu Huang1,*
...-injection of CpG adjuvant and somatostatin (SS) DNA vaccine can enhance the effect of the vaccine and promote the growth of animals. This study investigated the enhancing effect of the fusion type of CpG motifs on SS DNA vaccine and growth of ram lambs. In this study, we constructed the ptS/2SS-asd plasmid, pCpG plasmid and the CpG-SS fusion type recombinant plasmid named ptCS/2SS-asd. Twelve ram lambs were randomly divided into three groups, na...
Hao Gu*, Zhi Yang, Weiqiang Yu, Kan Xu and Yuan-fei Fu*
...s expected to be used as adjuvant medicine to treat caries.
...
Yan-Guo Han1,2, Jun-Hua Ye1, Qin-Tao Zhao1, Yong-Jie Huang2, Kai Li2, Yong-Fu Huang2 and Li Xu1,*
...l peptide and CpG adjuvant and delivered by attenuated Salmonella choleraesuis at weeks 0, 3 and 6 of the study, and rats in control group (Group C) was orally given empty vector vaccine. Blood samples were collected before primary immunization and at weeks 3, 5 and 7 after primary immunization and body weight of offspring were weighed at weeks 0, 2 and 4 after birth. Both SS-14 and SS-28 DNA vaccines induced humoral immune r...
Bibi Nazia Murtaza1, Mazhar Saeed Chaudry2, Shamaila Inayat Nadeem1, Muhammad Shahid Nadeem3 and Abdul Rauf Shakoori4,*
...lyse before the start of adjuvant therapy. Further studies needed to confirm the functional aspects of rs112445441 and other variants involved in NHL pathogenesis. 
...
Xuya Zhou1, Ying Liu1, Deqin Xu1, Jie Bao1, Yaru Cao2 and Yong Jin3*
...n of streptozotocin. The adjuvant-induced arthritis model was established by Paraffin/BCG. We used western blot to explore the protein expression levels of COX-2 and cytochrome P450 (CYP) 4A1. The expression of CYP4A1 was detected by immunohistochemistry. Enzyme-linked immunosorbent assay (Elisa) was used to detect the prostaglandin E2 (PGE-2), 20-Hydroxyeicosatetraenoic acid (20-HETE), endothelin 1 (ET-1) and B-type natriuretic peptide (BNP) in blood serum of...
Guangyu Wang, Xu Tian, Lintao Liu, Junyang Liu, Bo Zhang and Jingming Dong*
...so that effectiveness of adjuvant treatment can be improved.

 

...

Walaa ,S. Shabanaa;Bakr, A. Abdallaa; Eman, M. ELShalamia; I.M. Redab; M. A. Shalabyb

...the Montanide ISA 50 oil adjuvanted combined FMD/RVF
vaccine was tested in sheep and compares it with single vaccines either FMD or RFV alone.The
mean of antibody titers continued with the protective level till the 32 to 36th week post vaccination,
In single vaccination either FMD or RFV alone while till to 40th WPV in combined FMD/RVF
vaccine.In the final we can conclude that the use of Montanide ISA 50 as an oi...

Hiam M. Fakhry and Assem A.A.

... conducted to reveal the adjuvant's effect of zeolite on the immune response of oil adjuvanted trivalent Foot and mouth disease (FMD) vaccine in sheep. The objective of this study was to evaluate the efficacy of Zeolite (Zeolith UF) as an adjuvant using inactivated trivalent foot and mouth disease virus, to induce protection against foot and mouth disease. This study was conducted in three...

Walaa Abd El-Fatah S. Metwally1, Ismail M. Reda2, Ahmed A. El-Sanousi2, Mohamed Abd El-Khalek Ali2

... which has been added as adjuvant using reassortant AI strain H5N1. The prepared vaccines with Montanide ISA70VG varied in the HA units per dose and also differed in the ratios between two reassortant strains (Chicken and duck strains) of AI. The results of testing humoral immunity revealed that the prepared vaccines with Montanide ISA70VG containing 350 HA unit/dose were high in antibody titer than vaccines containing 256 and 100 HA unit/dose. The prepared va...

Afaf. A. Khedr*, Ekram Salama*, A. A. Samy**, Abdella, Y.A.*** and Suzan. K. Tolba***

...combined inactivated oil adjuvant vaccines against infectious coryza and infectious bronchitis diseases were prepared, one adjuvanted with mineral oil and the other with Montanide ISA-70. Evaluation of both vaccines were done by challenge test, haemagglutination inhibition test, ELISA test for Avibacterium Paragallinarum, and ELISA test, indirect haemagglutination inhibition test and ciliostasis score for infectious Bronchit...

1Mona A. El-Manzalawy, 2Abeer A. Boseila , 3Hanan M. El Zahed

... RVF vaccine Formulation adjuvanted by Peppermint oil (PMO) with Aluminum hydroxide gel (Alum gel) or without Alum gel, then ensure the safety and sterility of these vaccines and compare between them with the local RVF vaccine with Alum gel alone in sheep. This peppermint formulation including Alum gel adjuvant was the best of choice as it induced higher immunological enhancement. In addition, synergistic action was discover...

Amir M. Walaa1 ; Ausama Yousif1; Walid H. Kilany2; Magdy El-Sayed1; Ahmed A. El-Sanousi1

... vaccines with different adjuvants except for group 1 & 2. Group 1 was kept as unvaccinated control. At 28 days all groups were challenged with a dose of 105.5 EID50/bird of Variant 2 genotype (1494) IBV through oculo-nasal rout. Blood samples were collected randomly from 10 birds from each group at day 7, 14, 21, 28, 35 and 42 . Organs of 3 birds/each group were collected (trachea, bronchi and kidneys) for PCR at 3, 7 and 10 days post vaccination and post...

Maha A. El-Abhar1, Moustafa A. Elkady1, Khaled M. Ghanem2, Hussieny A. Bosila3

...hly economic
adjuvant in FMD vaccine for cattle.
...
Hanan Sayed1, Mohamed S. Madkour1, Hussein A. Hussein2, Mounir Mohamed Elsaft 3,
Marwa F. El saied3, Ahmed A El-Sanousi2
...2) by using montanide 71 adjuvant. The prepared vaccine was
injected intramuscularly in specific pathogen free and commercial layer chicken, the injection was
repeated as a booster vaccination one month after the first vaccination. Humoral immune response was
measured by HI for both H9N2 and LaSota viruses weekly for one and a half month after the booster
vaccination, then monthly until the end of duration. Chall...

Hanan Sayed 1, Mohamed S. Madkour 1, Hussein A. Hussein 2, Ahmed A El-Sanousi 2

...2) by using montanide 71 adjuvant. The prepared vaccine was
injected intramuscularly in specific pathogen free and commercial layer chicken, the injection was
repeated as a booster vaccination after a month from the first vaccination. Humoral immune response
for IBV was measured by ELISA weekly until about one month after the booster vaccination then
periodically till the end of duration. Challenge trial for both...

Youssef A. Elgharbawy1, Mervat M. Ali1, Ayman H. El-Deeb2, Mohamed A. Shalaby2

...ding equal volume of the adjuvants (Montanide ISA 50 and Montanide ISA 206 ) to the total quantity 3ml of the vaccine. The prepared vaccine tested for determination of immunological response against the vaccine with ISA 50 and ISA 206 adjuvants and the results was compared. The results of cellular and humoral immune response and the challenge test showed that using inactivated LSDV vaccine improved by Montanide ISA 206 eleva...

Abdelaziz A. Yassin, Ayman H. El Deeb, Abeer E. Mansour, Hussein A. Hussein

... reports the role of ail adjuvant in stimulating the cellular response for inactivated vaccine.

...
Soad, E. Elsayed 1, Mohammed A. Shalaby2, Ausama A. Yousef2, Abu bakr A. Agor1,
Sherouk E. Aly1
...Montanide ISA 206 is the adjuvant used in FMDV vaccine produced in Egypt which is
w/o/w microemulsion.
Methods: We aimed to increase the efficacy of the conventional formulation by converting the
microemulsion to Bio-compatible and stable Nano emulsion. Emulsification was applied through
ultrasonication of Montanide ISA 206 in the presence of Tween 80 as a detergent. The physical
characteristics were ...
Shourok E. Aly 1, Karim Zaki1, Soad E. Elsayed2, M. Elgamal2, Ausama A. Yousef1, Mohamed A. Shalaby1
...n aluminum hydroxide gel-adjuvanted inactivated Rift Valley
fever virus (RVFV) vaccine.
Objective: Herein, we aimed to enhance the immunogenicity of the conventional vaccine by
developing a smaller particle size vaccine formulation through ultrasonic dispersion of aluminum
hydroxide adjuvant.
Methods: This was achieved by ultrasonication of aluminum hydroxide gel in...

Muhammad Ehsan Safdar1*, Samra Ammara1, Muhammad Sikander Hayyat1, Shahbaz Hussain2, Muhammad Mansoor Javaid1, Abdul Rehman1 and Amjed Ali1

...n tank mixed addition of adjuvant (alkyl ether sulfate) at Research Area, University of Sargodha during spring 2015. The studies consisted of seven herbicide treatments, viz. lactofen complete dose (180 g a.i. ha-1) alone, lactofen complete dose + adjuvant, lactofen 80% dose alone, lactofen 80% dose + adjuvant, lactofen 60% dose alone, lactofen 60% dose + adjuvant<...
Tabinda Urooj1,*, Bushra Wasim1, Shamim Mushtaq2, Syed Nudrat Nawaid Shah1, Lubna Faisal3, Moazzam Ali2, Nabeela Rasheed1 and Syed Faizan Ali Rizvi4
...ived cancer treatment as adjuvant therapy, were included as treated (Group B) cases. Regarding patient’s demographic data, mean age of enrolled patients was found to be 48.26±10.74 years. The mean age of menarche was found to be 13.39±0.89 years. The calculated BMI (body mass index) was found to be 20.4. Majority of the enrolled subjects have acquired primary or basic education and most of them were married and housewives. For the reproduct...

Sana Shakoor, Tahir Rehman Samiullah, Naila Shahid, Abdul Qayyum Rao*, Aneela Yasmeen, Sana Tahir, Ayesha Latif, Saira Azam, Ahmad Ali Shahid and Tayyab Husnain

...ng with enhancer (ferund adjuvants) was done and produced antibodies in serum was detected by immunodot blotting assay. The appearance of the dot on spotted position confirmed the specificity of anti-HA antibodies. The titer of antibody produced in immunogenic mice was determined through ELISA. The highest geometric mean titer of HA-specific serum IgG, which was greater as compared to the control.

...

Ahmed Alazzouni1, Ashraf Al-Brakati2,*, Sherif Rabie1, Mohamed Gabry1 and Basmaa Hassan1

...herapeutic agents and as adjuvants to ameliorate histopathological changes associated with leukemia.

...

Mohamed A. Hussein1, Bahgat A. Abd El-Rehman2, Kareem A. Eldin2, Mohamed Fouad Mansour1* 

...osphate nanoparticles as adjuvants and evaluate their impact on biochemical, cellular and humoral immune response in sheep, The prepared vaccines were sterile, safe, emulsion is stable and validity within acceptable limit 0.02 EDF50/ml up till 15th month with AlP-NPsV and AlHV except Ch-NPsV that was valid up to 16th month. Lymphocyte count increased from the 1st week post-vaccination till the peak at 5th, 4th and 3th months in groups vaccinated with Ch-NPsV, ...

Jianmin Liu1, Jiancheng Zhang2, Peijuan Wang3,* and Moslem Lari Najafi4

... and sex hormones in the adjuvant treatment of ovarian function decline. Ninety patients with ovarian function decline were randomly divided into 3 groups: new addition congrong tusi pill treatment group (n=30 cases), diethylstilbestrol treatment group (n=30 cases) and the control group (n=30 cases). A comparative analysis is made on sex hormone levels, ovarian artery blood flow, ovarian volume and number, treatment efficacy, glucose metabolism, ovulation and ...

Rezqita Putri Pitaloka1, Kartiawati Alipin1, Mas Rizky A.A Syamsunarno2, Gemilang Lara Utama3, Ramdan Panigoro2, Ratu Safitri1* 

...nol extract (SWEE) as an adjuvant and substitute for iron chelator. The adjuvant test group was given 60 mg/kg BW iron dextran (ID), 75 mg/kg BW deferiprone (DFP), and 50 to 200 mg/kg BW SWEE, while the substitute group was administered with 60 mg/kg BW ID and DFP:SWEE with a gradient ratio. The ID was given orally for the first 14 days at intervals of 3 days, while DFP and SWEE were administered orally every day for 28 days...
Farah Abid1,2, Mohammad Saleem1,3*, Tahir Maqbool4, Tania Ahmed Shakoori5, Faheem Hadi4, Tahir Muhammad4, Saira Aftab4, Yasir Hassan7 and Shabana Akhtar4,6*

Xuan Liang, Rongfang Zhang, Yan Xu, Hui Yang, Li Guo, Feiyang Na and Donghai Liu*

...s) signaling pathways in adjuvant treatment of children with recurrent respiratory tract infections (RRTI). A total of 138 children with RRTI admitted to our hospital from May 2017 to May 2019 were selected as the study subjects and randomly divided into 3 groups with 46 cases in each group. The 3 groups were given conventional treatment. On this basis, control group A was given spleen aminopeptide adjuvant treatment, the co...

Dalia Zaafar1*, Heba M.A Khalil2, Soha Hassanin3, Mohamed R. Mousa4, Mona G. Khalil1 

...ore it can be used as an adjuvant agent to boost the therapeutic efficacy of chemotherapy in the treatment of many types of cancer.

Keywords | Pioglitazone; Cisplatin; Ehrlich solid carcinoma; Oxidative stress; VEGF; PPARγ; anticancer agents; tumorigenesis; biochemical indices 

...

Bilal Ahmed Shah1, Muhammad Avais1*, Jawaria Ali Khan1, Masood Rabbani2, Aftab Ahmad Anjum2, Muhammad Asad Ali2, Muhammad Awais1, Sohail Ahmad3 and Shahan Azeem2*

...in-inactivated montanide adjuvanted polyvalent mastitis vaccine (FMPV), formalin-inactivated aluminum hydroxide adjuvanted polyvalent mastitis vaccine (FAPV), and formalin-inactivated montanide plus aluminum hydroxide adjuvanted mastitis vaccine (FMAPV) were prepared after confirmation for antigenicity and immunogenicity and evaluated for safety, side effects, challenge protection assay, s...

Orooba Mohammed Saeed Ibrahim1*, Rawaa Saladdin Jumaa2, Nibras Zeyad Yahya1 

... in the market for novel adjuvants. The primary initial source of many medicines used to treat diseases today was plants. This study aims to ascertain the antibacterial activities of fruit Citrus aurantium and Citrus limonum juice on isolated Staphylococcus aureus, Sterptococcus pneumoniae and Klebsiella pneumonia. The antibacterial activity of fruit juice extract on bacterial strain was determined using macro-broth dilution, agar well diffusion methods and t...

Rana Mahmood Ahmad*, Orooba M.S. Ibrahim 

...he day of Freund’s adjuvant injection with SaZnONPs and piroxicam at doses 2.5 and 5 mg/kg B.W., respectively, while (Group 1) was positive control. Blood samples were obtained using cardiac puncture at three time points during the experiment: at the beginning (zero time) and on the 12th and 21st day. These samples were collected specifically for serum collection. In the present investigation, the administration of SaZnONPs to infected rats yielded notew...

Amal Hammad*, Shaaban Gadallah, Tarik Misk, Ahmed Mourad 

...nd Yintang as a sedative adjuvant in dogs. Five adult healthy mongrel dogs were randomly assigned to receive each of four sedation protocols at one-week interval. Hence, 4 groups were evaluated: XH (xylazine high: 1mg/kg of xylazine intravenously); XL (xylazine low: 0.25mg/kg of xylazine intravenously); XLL (xylazine low combined with laser acupuncture: 0.25mg/kg of xylazine intravenously with laser stimulation at GV20 and Yintang) and XLV (xylazine low combin...

Rizk, Sonia A.; El-garf, Eman M. and Talaat, Abeer A.

...e conduced to reveal the adjuvant's effect of Ginseng Stem-leaf Saponins (GSLS) on the immune response of gel adjuvanted Bivalent Foot and Mouth Disease (FMD) vaccinated calves. This study conducted in two calve groups; goup (A) vaccinated subcutaneously with bivalent Alhydragel adjuvanted (30) % FMD vaccine, while group (B) vaccinated subcutanously with bivalent FMD vaccine

Ebied, Eman M.; Salem, Zeinab T. and Al-Imam, Hemmat S.

...) Dextran solution as an adjuvant. It was found that mares developed high haemagglutinating inhibition (HI) titre at 3 weeks post vaccination and remain with protective level up to 6 months, while their foals had moderate (HI) antibodies within  48 hours post colostral suckling, the HI antibodies rise to a level similar to their dams within one to two weeks then persisted with considerable level up to 6 months of their age.

...

Sonia, A.M.; El. Sanousi, A.A.; Saber, M.M.; Daoud, A.M.; Samira, E.K.; and Ismail I.

... by using four different adjuvants which are: Montanide ISA 206, Quil A saponine, Ginseng extract and Montanide IMS 3015. These vaccines were compared with Al (OH) 3 gel vaccine and tested in calves. The obtained results revealed that the duration of immunity elicited by gel FMD vaccine was shorter than oil adjuvanted FMD vaccines. Results also indicated that vaccine emulsified with Montanide IMS 3015 could elicit the best p...

Abdel Razek,B.Omar*and Magda M.Sayed**

...ncomplete Freund’s adjuvant. Identity tests were applied by agar gel precipitation test (AGPT) and indirect fluorescent antibody technique (IFAT). Serum neutralization test (SNT) and ELISA tests were applied for titration and evaluation of the hyperimmune sera before conjugation with fluorescent. Total protein concentration of the prepared LSDV antisera was 0.8g/dl. Separation of anti-LSDV immunoglobulins 1gG were done using ammonium sulphate followed by...

Arwa H. El -Naggar, Nirmeen G. Shafiek, and M. S. Wassel.

... and incomplete Freund's adjuvant and without adjuvant in successive doses into susceptible guinea pigs. Serum neutralization (SNT), ELISA and indirect fluorescent assay (IFA) tests were used to evaluate the hyper immune serum Before conjugation with fluorescent. The antibody titer reach 32 by SNT and 5984 by ELISA and identified by IFA to be Bovine rota. Separation of anti B. Rota immunoglobulins IgG "were done using a...

Salema Lafta Hassan1*, Taghred Jabbar Humadai1, Sabrin Ibraheem Mohsin

...culation with 0.1 ml oil adjuvant vaccine (OAV) of Pasteurella multocida twice a week for two weeks. The third group was vaccinated by I/P with 0.1 ml oil adjuvant vaccine (OAV) of Pasteurella multocida in two doses separated by two weeks. The fourth group (G4) was assigned the role of negative control at this time, the 28th day after immunization for work, serum samples were taken for passive hemagglutination test (PHA); Im...
Abdul Asim Farooq1,2*, Muhammad Arif Khan1, Hamid Akbar1
Muhammad Ashraf3, Saima Inayat4, Muhammad Usman Saleem5
Saeed Murtaza2, Maqbool Hussain Shah2 and Muhammad Arshad Javid5
... an efficient biological adjuvant for healing long bone fractures. BMA comprises bone mesenchymal stem cells. This study aims to assess the potential of autologous BMA on metacarpal and metatarsal fracture of Beetal goats presented at the surgery clinic of the University of Veterinary and Animal Sciences Lahore, Pakistan. Beetal goats were selected (n=20), and divided into four different groups. The first group was designated as bone plating with bone marrow a...

Saira Jabeen1, Muhammad Avais1*, Jawaria Ali Khan1, Kamran Ashraf2, Aftab Ahmad Anjum3

...each employing different adjuvants. To ensure the quality and safety of these vaccines, quality control assessment was carried out. The efficacy evaluation of the vaccines was performed on 24 locally bred rabbits, divided into 4 groups. Each group received a different type of vaccine, with booster doses at 15-day intervals following the priming. Serum samples were collected biweekly over a 60-day period, and the antibody titers were measured using IHA test. Ge...

Sura Ayed Radam*, Inam Badr Falih

...volume of Freund’s adjuvant, 2 doses, 14 days intervals, 2nd group was immunized s/c with Concanavalin-A Con-A (1.5 mg/ml), 3rd group was immunized with mixed S. pyogenes Ag. and Con-A, 2 doses, 14 days intervals, and 4th group was given Phosphate buffer saline (PBS) as negative control group. Results Analysis of cellular and humeral immunity recorded higher concentration of Immunoglobulin G IgG, Tumer Necrosis Factor alpha TNF-a and Interleukin 10 IL-10...

Maysaa A. Jumaah*, Ali A. Al-Iedani 

... oil) to current vaccine adjuvants (alum) using inactivated antigens (sonicated and whole-killed bacteria) of E. coli. We also aimed to reveal tthe nature of host innate and adaptive immune responses by measuring the extent of their effects on the level of pro-inflammatory (interferon γ and interleukins (IL)-12) and anti-inflammatory cytokines (IL-4 and IL-10) and innate immune responses (total white blood count, phagocytosis and respiratory burst). Our ...

Ibrahim Ali1,2, Hanan Mohamed Fathy Abdien1*, Wael Kamel Elfeil1, Mohsen Mohamed Zaky ElDemerdash1, Mohamed Ali Zain El-Abideen2,3, Walid Hamdy Kilany2,3

...ssical Oil excipients as adjuvants. We have formulated a novel monovalent experimental vaccine against the recent circulated HPAI H5N8 clade 2.3.4.4b viruses using following adjuvants (MONTANIDE™ GEL 01 PR, MONTANIDE™ IMS 1313, and MONTANIDE™ ISA70 to evaluate their impact on the vaccine efficacy, potency, with special focus to humoral and cell mediated immune response. Seven groups of specific pathogen-fre...

Mostafa R. Zaher1,2*, Amir A. Shehata1, Azza M. El Amir2, Naglaa M. Hagag1, Reham H. Tammam3**

...nts. Besides, the use of adjuvants and delivery systems to boost vaccine performance was also reviewed. Furthermore, we explore state-of-the-art methodologies such as exosome-based vaccines, cell-free protein synthesis systems, self-amplifying RNA vaccines, and the application of artificial intelligence (AI) and computational biology in vaccine design. These methodologies may offer solutions for the challenges posed by the foot-and-mouth disease virus’s ...

Diaa El-Boraey1, Mousa A. Ayoub1, Wael K. Elfeil2*, Said Ibrahim Fathalla3, Ibrahim Said Abu-alya3, Mohamed Abaza4, Ahmed R. Gado5, Mobarez A.A.6, Nehal K. Alm Eldin6

...inactivated oil in water adjuvant avian influenza vaccine (H9) in broiler chickens. Four groups of chickens were established as follow: a negative control, a positive control (vaccine only), a group receiving the vaccine with amikacin, and a group receiving the vaccine with gentamicin. Hemagglutination inhibition (HI) titers were measured to assess antibody responses in addition to performance parameters. The obtained results demonstrated a reduction in antibo...

Advances in Animal and Veterinary Sciences

November

Vol. 12, Iss. 11, pp. 2062-2300

Featuring

Click here for more

Subscribe Today

Receive free updates on new articles, opportunities and benefits


Subscribe Unsubscribe