The selectivity of the FBBB test was evaluated by analyzing 5 other cannabinoids, herbs, spices, essential oils, tobacco, and hops. None of these substances produced color like that of FBBB + THC nor fluorescence observed. For the colorimetric calibration experiments, it was shown that when the ratio of THC:CBD is above 1, a red color forms indicating that there is marijuana present. These experiments also found that the absolute LODs for THC on the PSPME substrates was as low as 500 ng, which is significantly lower than the LOD for the D-L test . The THC LOD for the 4-AP test is not currently known but expected to be >500 ng. This study demonstrates that the FBBB test is very selective and sensitive for THC, forming a red color and an intense fluorescence that can be distinguished from other chromophores. In addition, this chromophore is long lasting, allowing the color and fluorescence to be observed long after the test is performed. This long-lasting color is attributed to the nature of the FBBB being a diazonium salt, which are known to be stable and even used to form dyes in textiles . One limitation that was discovered for the FBBB test is that the reagent is not stable at room temperatures over more than a few days, losing its color and producing no reaction with THC or CBD. The FBBB reagent and the preloaded FBBB substrate were stable in the refrigerator/cooler for at least 45 days. The temperature instability is not ideal for field work since a kit using the Fast Blue BB test would likely be exposed to temperatures above 4 ◦C. For this reason, future work will focus on determining a method to maintain the FBBB stable at ambient temperatures. Malaria is a tropical parasitic disease of plasmodium notorious for the increased death among children and pregnant women especially in the poor country of the world. The severity of the disease is based on the species of the plasmod- ium . Malaria has always been a health threat that has relentlessly taken life for millennia.The use of herbal alternatives was necessitated by the development of an effective drug for the global control of this life-threatening disease.
As a result, there is an urgent need to develop antimalarial drugs from chemical, synthetic, and natural sources to mitigate the effects of this tropical disease. However, in the design of antiparasitic drugs, such as in the case of the plasmodium parasite, re- searchers tend to consider bioactive agents with the potential to alter cellular processes that are harmful to these organisms by binding on strategic target sites. Cannabis extract is renowned for its therapeutic properties due to the presence of a high proportion of antioxidant phytochemicals, particularly coumarins.Coumarin and its synthetic derivatives have been shown to inhibit the proliferation of malarial parasite by targeting microtubule dynamics.Tubulin inhibitors, such as coumarin, impede malarial parasite proliferation and growth.The vital role of microtubules in cellular processes on the erythrocytic cycle of P. falciparum, include the following: merozoite formation,invasion of erythrocytes by daughter merozoites, and nuclear division. Furthermore, the protein biosynthesis function of the microtubule has made it a desirable drug target site. The clinical symptoms and manifestations of malarial infection are during the parasite’s erythrocytic stage which is also the asexual period of the development of the parasite.The ‘silent phase,’ also known as the pre-erythrocytic stage of the infection, has no symptoms and affects only a few hepatic cells . One of the aspirations of the African Union’s Agenda 2063 is for a prosperous Africa, based on inclusive growth and sustainable development goals, attainable through a high standard of living, quality of life, and well-being for all citizens through a focus on health and nutrition. As a result, one of the identified challenges to this goal is malaria infection, which is endemic in Africa and has been linked to the majority of deaths among pregnant women and children under the age of five. This eventually led to poverty, which is blamed for Africans’ low standard of living. This current study could be used to develop a low-cost and effective malaria treatment option for the African continent. This research investigated the anti-malarial properties of cannabis grow equipment in vivo as well as possible inhibitory effect of a cannabis GC–MS compounds on the plasmodium parasite’s microtubule target protein in silico. The anti-plasmodial function of medicinal plants is due to the presence of bioactive ingredients in the plants, which have been reported to inhibit growth or cell replication.
These bioactive ingredients exhibit this therapeutic property by bind- ing to protein receptors or by blocking the binding site of an enzyme in a critical pathway for the survival of the organism. Symptoms of plasmodium infection in mice include anemia, hypoglycemia, weight loss, and a drop in body temperature.The positive control group showed an increase in weight because the mice in this group were not infected but given food and water only . The negative control group showed a decrease in weight compared to the standard drug which showed an increase in weight . Also, the mice treated with 100 mg/kg BW and 200 mg/kg BW respectively showed a decrease in weight and the mice treated with 400 mg/kg BW showed an increase in weight . There was a significant decrease in the percentage parasitaemia level in the mice treated with the extract on the 10th and 12th day post infection treatment compared to the infected untreated mice . The standard drug, chloroquine treated mice however showed a significant decrease in the percentage parasitaemia level from the 4th day post infection treatment compared to the mice in the infected untreated mice . On the 10th and 12th days of post-infection, mice treated with extract had between 80 and 96 percent inhibition in Plasmodium berghei NK-65 in infected mice compared to mice infected and treated with chloroquine which had 100 percent inhibition at the same time . Certain criteria, however, are required to track the therapeutic progress of drugs or medicinal plants. The increase in percentage weight change could be attributed to the therapeutic effect of cannabis extract.Also the therapeutic efficacy of antimalarial drugs is determined by their ability to rid the host organisms of parasites in the blood or the tissue. Furthermore, Plasmodium parasite merozoites invade red blood cells, where they feed on hemoglobin and multiply, thereby increasing the parasite’s population.If no antimalarial agent is applied, the parasite multiplies rapidly in the blood, increasing the parasite count by up to 20 times in 48. The parasitaemia level in the blood is an important criterion for determining the severity of malaria infection or an indicator of antimalarial drug therapeutic efficacy. The lower percentage of parasitaemia reported in this study could be due to the extract’s antiplasmodial activity. The percent- age inhibition of plasmodium parasites is a measure of how well an antimalarial drug prevents them from multiplying and growing. This study found that a high concentration of the antimalarial bioactive compound inhibited parasite growth in a dose-dependent manner, implying that a higher dose of the extract contained a higher concentration of the antimalarial bioactive compound. Furthermore, isolated cannabis compounds were found to have mild antimalarial activity in vitro with 4-acetoxycannabichromene, 5-acetyl-4-hydroxycannabigerol, and −1 _ S hydroxycannabinol . In another study, Akinlola et al. reported mild antimalarial activity in infected mice fed cannabis compounded feed . The use of crude extract in this study, on the other hand, results in a high antimalarial activity. Moreover previous researches have linked disease condition and therapeutic effect to changes on some hematological parameter.
There was a significant decrease in the WBC volume in the positive control, the groups administered 400 mg/kg of the extract and CQ treated groups compared to the normal control while groups administered 200 and 400 mg/kg body weight of the extract showed a significant increase . There was a significant decrease in the RBC level in the mice in the positive control and significant increase in the treatment groups and standard drugs compared to the normal control . There was no significant change in the hemoglobin concentration in all the groups compared to the normal control . There was a significant increase in the HCT level in the groups treated with 100 and 400 mg/kg body weight of the extract compared to the normal control . There was no significant change in the treatment groups, positive control and standard drug compared to the normal control . Mean Corpuscular Volume, and Mean Corpuscular Hemoglobin did not show significant change across groups compared to the normal control . MCHC showed significant increase across the groups compared to the normal group . There was a significant decrease in platelet across the groups compared to the normal control . No significant change was observed in the level of lymphocyte in all the groups compared to the normal control . Mixed Cell Count showed a significant increase in the positive control. There was a significant increase in level of plasma Platelet Crit Circulation in the infected and treatment groups compared to the normal control.Because the parasite’s primary target is RBC, the decrease in RBC can be attributed to either the invaded parasite destroying RBC or the spleen rapidly removing parasitized RBC from circulation.The normal RBC volume in the treated groups supported the antiplasmodial activ- ity by the cannabis extract. Red cell volume is measured by the haematocrit . The parasite’s invasion of the RBC and subsequent destruction of the RBC will eventually overwhelm the RBC production rate, resulting in a lower haematocrit. The HCT volume in the treated group was normal because there was no or a low level of parasitemia, which could destroy the RBC. The infection did not affect the levels of HGB, MCV, MCH, or MCHC, which may be attributed to some physiological compensation by the variant red blood cell . Platelet reduction is linked to malaria, mobile grow system and thrombocytopenia may be a sign of plasmodium infection.In response to increased parasitized red cells, platelets protect the erythrocyte by releasing platelet factor 4 molecules, as was demonstrated in previous studies. The plasmodial activity of platelet factor 4 molecules has been documented . The platelet’s antiplasmodial activity could kill the platelet, which could result in a decrease in PCT. The PCT was not affected in this study, which was in contrast to the findings of Asare et al., who reported a decrease in PCT . Platelet distribution width is a measure of platelet volume variability, and it increases when platelet anisocyto- sis is present. It is used to explicitly quantify platelet size variability, changes with platelet activation, and represents platelet morphology heterogeneity. In contrast to Yusuf et al. who observed increased PDW due to increased development of larger platelets as mitigation for excess platelet destruction in malarial conditions, this study found no increase in PDW in infected mice as compared to non-infected or treated infected mice.
The neutrophil is abundant in the peripheral blood, where it performs phagocytic action against antigens or microorganisms in the same environment as parasitized red blood cells. The decrease in neutrophil levels recorded in this study backs up a previous study that found a decrease in leukocyte levels in the blood stage of plasmodium infection and an increase in leukocyte levels in the liver stage of infection.The presence of blood parasitaemia is indicated by a decrease in the number of leukocytes in the blood. The most widely studied markers of platelet activation are mean platelet volume.The infection by plasmodium parasite in the present study did alter the level of MPV; this may imply that the infection did not induce increased development of new megakaryocytes. Furthermore, the antiplasmodial effect observed in this study demonstrated the extract’s therapeutic efficacy against plasmodium infection. This suggests that the extract’s compounds are acting synergistically to inhibit plasmodium parasite growth and proliferation in infected mice, possibly through molecular interactions between these compounds and some target macromolecules such as proteins. Also Bioinformatics techniques such as molecular docking and drug-likeness prediction are commonly used to predict and rank the potentiality of a compound in drug discovery. The in silico methods are less time consuming and effective at screening a large number of compounds. Literature abounds to justify the use of in silico methods for the screening of plant compounds targeted at the manage- ment of several human diseases. Such studies often probe into the interactions between plant compounds and protein tar- gets to identify those with favourable binding characteristics. In this study, we combined molecular docking of compounds from Cannabis crude extract with Plasmodium tubulin to predict drug-likeness of the compounds.