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  Original Article  Clinical Efficacy of a West African  Sorghum bicolor  -BasedTraditional Herbal Preparation Jobelyn ShowsIncreased Hemoglobin and CD4 + T-Lymphocyte Counts in HIV +  Patients Godwin I. Ayuba, MD, 1 Gitte S. Jensen, PhD, 2 Kathleen F. Benson, PhD, 2 Ademola M. Okubena, BA, 3 and Olajuwon Okubena, FCMA 3 Abstract Objectives:  The purpose of this study was to evaluate a traditional herbal preparation, Jobelyn,   for its effectson anemia and CD4 +  T-cell counts in human immunodeficiency virus–positive (HIV + ) patients in Nigeria. Design:  An open-label pilot study involving 10 confirmed HIV +  patients who were not receiving antiretroviraltherapy (ARVT) was performed, in which the patients consumed Jobelyn for 8 weeks, at a dose of 500mg twicedaily. The pilot study was followed by a controlled trial involving 51 patients, all confirmed HIV + , where thepatients with CD4 +  T-cell counts below 350 cells/ l L were receiving ARVT. The eight patients with baselineCD4 +  T-cell counts above 350 cells/ l L received Jobelyn. The remaining patients who all received ARVT wererandomized to ARVT alone versus ARVT +  Jobelyn for 12 weeks. Results:  Patients receiving ARVT showed a statistically significant improvement in their CD4 +  T-cell countsacross the 12-week study period (  p < 0.01). Patients receiving ARVT +  Jobelyn showed a faster improvement,reaching a high level of statistical significance compared to baseline already at 6 weeks (  p < 0.001), and remainedhighly significant at 12 weeks (  p < 0.001). Conclusions:  This is the first controlled study conducted to evaluate efficacy of Jobelyn on immune status inHIV +  patients. The data suggest that consumption of Jobelyn contributed to improved hemoglobin levels andincreased CD4 +  T-cell counts in Nigerian HIV +  patients. Further studies are needed to examine similar effectsin other populations, and to elaborate on the underlying mechanisms, specifically, whether the consumption of  Jobelyn supported multiple aspects of bone marrow function. Introduction T here is a worldwide recognition  of the vital roles of the need for integrated interventions, including herbalmedicines, as a necessary tool in global health care, particu-larly in third-world countries with poor access to pharma-ceutical therapeutic strategies that are widely used inindustrialized countries. With a focus on the human immu-nodeficiency virus (HIV), the World Health Organizationencourages the use of available resources for primaryhealth care. It further recommends that governments givehigh priority to the use and incorporation of proven tradi-tionalremediesintoacomprehensivehealthcare. 1 Despitethewell-developed pharmaceutical industry in industrializedcountries, the standard-of-care in most third-world countriesplaces emphasis on the use of local resources such as tradi-tionally used medicinal plants. Jobelyn  is a natural product developed from tropicalplants found within the Nigerian flora. Toxicological evalua-tion in laboratory animals has shown a satisfactory safety andside-effects profile. 2 It has been found to induce rapid andmarked improvementinanemia. 3–6 The suggested mechanismof action was an immune-boosting effect, which is interestingin light of the presence of antiviral peptides in some parts of the sorghum plant, 7 as well as unique polyphenol compoundsidentified in Jobelyn. 8,9 Evidence  in vitro  for many biologicalproperties of Jobelyn was recently published. The data includeimmune modulation, activation of natural killer cells involved 1 Department of Pathology, 68 Nigerian Army Reference Hospital, Yaba, Lagos, Nigeria. 2 NIS Labs, Klamath Falls, OR. 3 Health Forever International, Ikeja, Lagos, Nigeria. THE JOURNAL OF ALTERNATIVE AND COMPLEMENTARY MEDICINEVolume 0, Number 0, 2013, pp. 1–4 ª  Mary Ann Liebert, Inc.DOI: 10.1089/acm.2013.0125 1  in antiviral immune defense actions, and up-regulating of theproduction of antiviral chemokines. 10  Jobelyn’s effects on anemia 3–6 lead to the speculationwhether the effect on red blood cell production as well asproduction of other cell types may be positively affected.Limited use in uncontrolled trials in antiretroviral-naı¨ve HIV-infected subjects as well as patients on highly active anti-retroviral therapy (HAART) has suggested some improve-ment in several hematological parameters and quality of life. 11 HAART uses a combination of reverse transcriptase inhibitorsand protease inhibitors and is the current standard drugtreatment of HIV/acquired immune deficiency syndrome(AIDS). Appropriate and consistent use of HAART can helpreduce viral load and allow for some immune system recon-stitution, including an increase in CD4 +  T-cell counts. How-ever, even with heavy subsidy, HAART is neither accessiblenor affordable to a majority of people living with HIV/AIDSin third-world countries. Use of HAART is also limited bypotential toxicity and side-effects, as well as development of resistant strains of HIV. Only patients whose biologic profilesfit the eligibility criteria are typically treated with HAART.These limitations emphasize the need for continued search forviable alternatives or adjuncts to HAART.Increasing use of Jobelyn by many HIV +  patients andpeople with anemia justifies further evaluation of its efficacyfor support of production of lymphocytes and red blood cellsin people with bone marrow suppression. This study wastherefore designed to address these questions. Two clinicalpilot studies were performed on patients with HIV in Lagos,Nigeria. An initial pilot study involved 10 HIV +  patientswho also suffered from anemic conditions. A subsequentcontrolled study continued to evaluate the effects of CD4 + T-cell counts in HIV +  patients, some of whom also receivedantiretroviral treatment. The controlled study aimed atevaluating the role (if any) of Jobelyn in the management of HIV in the presence or absence of HAART. Methods Nutritional supplement  The consumable nutritional supplement Jobelyn wasgrown, harvested, and manufactured by Health ForeverProducts, Ikeja, Lagos, Nigeria, using seed stock from a re-cently domesticated variant of West African  Sorghum bicolor .One lot was used for the open-label pilot study, and a secondlot was used for the controlled trial. Both lots had similarchemistry profiles showing a very high content of polyphe-nols, particularly apigeninidin, which was present at ap-proximately 4% of the dry weight of the botanical material(Table 1). This level is consistent with other lots tested forimmune-modulating activities. 10 Trial 1 Ten patients (4 men and 6 women, age range 23–49 years)with confirmed HIV + , a reduced number of CD4 +  T Table  1.  Concentration of Five Polyphenol Marker Compoundsin the Two Product Lots Used for the Two Clinical Studies Trial Lot number Unit Apigeninidin Luteolinidin Apigenin Luteolin Naringenin Trial 1 06-4812  l g/g 38,650 323 6020 585 230Trial 2 09-0590  l g/g 47,380 890 6460 673 166 The quantification of these five polyphenol marker compounds was evaluated by the highly sensitive LC/MS-MS methodology, and theirstructures were confirmed by CAD-MS/MS analysis. The testing was performed by Brunswick Laboratories, Norton, MA. Table  2.  Data from Trial  1  Involving  10  Human Immunodeficiency Virus–PositivePatients Showing  CD4 +  T-Cell Counts and Hemoglobin Levels CD4 +  T-cell counts a  Hemoglobin  b Vol. no. Gender Age ARVT JOB Week 0 Week 4 Week 8 Week 0 Week 4 Week 8 1 M 25 No Yes 399 617 708 11.6 12.6 13.32 M 49 No Yes 656 704 824 12.3 12.8 13.03 M 35 No Yes 452 662 724 12.1 12.8 13.04 M 31 No Yes 518 530 560 12.0 13.0 14.05 F 23 No Yes 352 390 564 10.3 12.0 12.56 F 45 No Yes 460 617 669 10.6 11.0 11.07 F 31 No Yes 499 550 550 10.3 11.2 11.48 F 47 No Yes 830 1082 1203 9.8 10.3 11.29 F 31 No Yes 350 461 475 8.9 9.6 10.610 F 40 No Yes 385 458 622 10.2 11.3 12.0Average 490 607 690 10.8 11.7 12.2SEM 47.64 61.29 65.51 0.36 0.37 0.36  p -value c < 0.01  < 0.001  < 0.001  < 0.001 a CD4 +  T-cell counts are shown as cells/ l L whole blood.  b Hemoglobin levels are shown as g/dL. c Data analysis utilized the within-subject two-tailed paired  t -test to compare each person’s changes for Week 4 and Week 8 to baselinevalues.ARVT, anti-retroviral therapy; JOB, Jobelyn,  which was consumed at a dose of 500mg twice daily; SEM, standard error of mean. 2 AYUBA ET AL.  lymphocytes, who did not qualify for antiretroviral therapy(ARVT), and who consumed Jobelyn, were followed throughthe Police Hospital in Lagos, Nigeria for 8 weeks. Potentialstudy participants were regular patients of Dr. N. U. Eguaeand were invited to participate by use of written informedconsent. The number of CD4 +  T lymphocytes and the he-moglobin level were evaluated at baseline, 4, and 8 weeks. Trial 2  Based on these pilot data, a controlled trial was performedat the Clinical Research Division of the Military Hospital,Ikoyi, Lagos, Nigeria. Fifty-one patients (28 men and 23women, age range 18–67 years) were enrolled in the studyupon written informed consent, as approved by the IkoyiMilitary Hospital Institutional Review Board. Potential studyparticipants were regular patients of Dr. Ayuba, and invitedto participate, by use of written informed consent. All studyparticipants who at the time of screening had CD4 +  T-cellcounts below 350 cells/ l L were treated with ARVT, whichconsisted of nevirapine (200mg), lamivudine (150mg), andstavudine (40mg) twice daily. All 51 patients who partici-pated in this clinical trial were monitored for CD4 +  T-cellcounts at baseline, 6, and 12 weeks. Eight patients who at thetime of screening had a CD4 +  T-cell count at 350 or higherwere ineligible for ARVT, and received Jobelyn for the sameduration of 12 weeks. The remaining 43 patients were ran-domized into two groups, where one group of 16 people wasonly treated with ARVT, and the other group of 27 peoplewas treated with ARVT and also consumed Jobelyn daily for12 weeks. There was no significant difference in the baselineCD4 +  T-cell counts between these two groups. Data wereanalyzed using ‘‘within-subject’’ analysis using the pairedtwo-tailed  t -test, as well as ‘‘between-groups’’ analysis, usingthe unpaired two-tailed  t -test. Results Trial 1 A statistically significant increase in CD4 +  T lymphocytes(  p < 0.01) and hemoglobin (  p < 0.001) was observed within 4weeks, using the paired two-tailed  t -test. Among the 10 par-ticipants, everyone showed an improvement in both CD4 +  T-cell count and in hemoglobin levels (Table 2). All 10 patientshad hemoglobin levels below normal for their gender at baseline. All of them showed an improvement in hemoglobinlevels. After 8 weeks of consuming Jobelyn, all 10 patients hadhemoglobin levels near the lower normal range. Trial 2  Among the 51 patients participating in this controlled trial,43 patients had baseline CD4 +  T-cell counts at or below 350cells/ l L, and received ARVT.As expected, people on ARVT showed a statistically sig-nificant improvement in their CD4 +  T-cell counts across the12-week study period ( n = 16,  p < 0.01) (Fig. 1). However,patients receiving ARVT +  Jobelyn showed a more robustimprovement, which reached a high level of statistical sig-nificance compared to baseline already at 6 weeks ( n = 27,  p < 0.001), and remained highly significant at 12 weeks(  p < 0.001) (Fig. 1). Analysis comparing the two groupsshowed that the CD4 +  T-cell counts in the group receivingARVT +  Jobelyn was significantly higher than the group re-ceiving ARVT alone, both at 6 and 12 weeks (  p < 0.001).Within the study population of 51 people, only 3 patientshad lower CD4 +  T-cell counts after the 12-week study than atstudy baseline; all 3 patients received ARVT, and 2 of them alsoconsumed Jobelyn. These three people experienced concurrentinfections (tuberculosis, pneumonia) during the study. Patients’appetite and weight increased and there was improvement intheir general well-being. The maculopapular skin rash of threeof the patients who had herpes zoster remitted.The third group of patients (i.e., those eight people whoreceived Jobelyn only, since they were not eligible for ARVTdue to a higher CD4 +  T-cell count at baseline) also showedan improvement during the weeks of Jobelyn consumption(Fig. 1). This improvement was statistically significant from baseline after 12 weeks (  p < 0.01). Discussion The core result presented here is the improvement inCD4 +  T-cell numbers over time in the two groups that FIG. 1.  The changes in CD4 +  T-cell counts from Trial 2 (acontrolled study with 51 HIV +  patients, where HIV +  statuswas confirmed by electrophoresis) are shown as the groupaverage – standard error of the mean. Improvements wereseen for all three groups: Eight patients who started thestudy with CD4 +  T-cell numbers higher than 350 cell/ l L,and were not eligible for antiretroviral therapy (ARVT)consumed Jobelyn (JOB) for 12 weeks. Improvementreached statistical significance at 12 weeks (  p < 0.01). Forty-three patients with baseline CD4 +  T-cell counts lower than350 cells/ l L were eligible for ARVT, and were randomizedto either ARVT alone or ARVT and Jobelyn. There was nostatistical significance in the CD4 +  T-cell counts at baseline between these two groups. The group receiving ARVTalone showed the expected improvement in CD4 +  T-cellcounts, which reached statistical significance already at 6weeks (  p < 0.01), and continued to improve at 12 weeks(  p < 0.001). The group that received ARVT and also con-sumed Jobelyn showed a more robust improvement, whichreached a high level of significance already at 6 weeks, andremained highly significant at 12 weeks (  p < 0.001). Dataanalysis utilized the within-subject two-tailed paired  t -testto compare each person’s changes for week 6 and week 12compared to baseline values, and also the between-groupsunpaired  t -test to compare the data for the ARVT +  JOB + and the ARVT +  JOB- groups at 12 weeks. Levels of statis-tical significance are indicated by asterisks: **for  p < 0.01,and ***for  p < 0.001. JOBELYN IMPROVES CD4 +  T-CELL COUNTS IN HIV +  PATIENTS 3  consumed Jobelyn when compared to the group treated withconventional antiretroviral treatment only. An additionalobservation from the initial pilot study is the improved he-moglobin levels seen in HIV +  patients consuming Jobelynfor 8 weeks.The parallel increase in hemoglobin and CD4 +  T-cellcounts seen in the pilot study suggests that consumption of  Jobelyn may support several aspects of bone marrow func-tion. The data from the controlled study involving 51 peoplesuggest support of T-cell production. The primary mecha-nism likely includes support of bone marrow function sinceanemic conditions were also reduced, possibly combinedwith secondary mechanisms of enhanced immune defenseagainst viral infection. In addition, an antiviral effect may besuggested, based on the observations that Jobelyn providesseveral aspects of antiviral immune support  in vitro,  whichincludes activation of natural killer cells and production of chemokines MIP-1a, MIP-1b, and RANTES. 10 Future clinical studies need to address whether Jobelyn hassimilar effects in populations with different demographics,diet, and lifestyle, as well as different prevalence of concom-itant infectious diseases. Also, the suggested mechanisms of action may be further evaluated through testing of serumcytokine/chemokine profiles, including growth factors withknown effects on bone marrow function, and antiviral che-mokines. These effects by Jobelyn may suggest a role for itsuse in broader health management, and not limited to HIV orHIV-associated anemia alone. Clinical trials are currently on-going to examine the effects of Jobelyn on anemia in non-HIV +  study populations. Given the significant effects seenover a relatively short study period in a West African popu-lation, consideration should be given to expanding its use inother countries with different health challenges, to help aug-ment health improvements in acute and chronic viral illnesses. Acknowledgments The two studies presented here were conducted at thePolice Hospital and the Military Hospital, both in Lagos,Nigeria. The data analysis was performed at NIS Labs,Oregon, United States. The studies were sponsored byHealth Forever International, Inc. Disclosure Statement G.S. Jensen and K.F. Benson are associated with NIS Labs,and Colonel G.I. Ayuba was employed as staff physician atthe Military Hospital at the time this study was conducted.None of these people have any commercial interest in thissubject matter. A. and O. Okubena are directors at HealthForever International, Inc., the sponsor of this work. References 1. WHO, UNAIDS, and UNICE, 2011 Global HIV/AIDS re-sponse: Epidemic update and health sector progress towardsuniversal access, World Health Organization, Geneva, Swit-zerland, 2011.2. Eniojukan JF, Aina BA. Toxicological profiles of commercialherbal preparation Jobelyn  . Int J Health Res 2009;2:369–374.3. Erah PO, Asonye CC, Okhamaf AO. Response of Trypano-soma brucei brucei–induced anaemia to a commercial herbalpreparation. Afr J Biotechnol 2003;2:307–311.4. Ogwumike OO. Hemopoietic effect of aqueous extract of theleaf sheath of Sorghum bicolor in albino rats. Afr J BiomedRes 2002;5:69–71.5. Oladiji AT, Jacob TO, Yakubu MT. Anti-anaemic potentialsof aqueous extract of Sorghum bicolor (L.) moench stem bark in rats. J Ethnopharmacol 2007;111:651–656.6. Akande IS, Oseni AA, Biobaku OA. Effects of aqueous ex-tract of Sorghum bicolor on hepatic, histological andhaematological indices in rats. J Cell Animal Biol 2010;4:137–142.7. Camargo Filho I, Cortez DA, Ueda-Nakamura T, NakamuraCV, Dias Filho BP. Antiviral activity and mode of action of apeptide isolated from Sorghum bicolor. Phytomedicine2008;15:202–208.8. Geera B, Ojwang LO, Awika JM. New highly stable dimeric3-deoxyanthocyanidin pigments from Sorghum bicolor leaf sheath. J Food Sci 2012;77:C566–C572.9. Kayode APP, Nout MJR, Linnemann AR, Joseph D. Houn-houigan JD, Berghofer E, Siebenhandl-Ehn S. Uncommonlyhigh levels of 3-deoxyanthocyanidins and antioxidant ca-pacity in the leaf sheaths of dye sorghum. J Agric FoodChem 2011;59:1178–1184.10. Benson KF, Beaman JL, Ou B, Okubena A, Okubena O, Jensen GS. West African Sorghum bicolor leaf sheaths haveanti-inflammatory and immune-modulating propertiesin vitro. J Med Food 2013;16:230–238.11. Awodele Olayemi SO, Adeyemo TA, Sanya TA, Dolapo DC.Use of complementary medicine amongst patients on anti-retroviral drugs in an HIV treatment centre in Lagos, Ni-geria. Curr Drug Saf 2012;7:120–125. Address correspondence to: Olajuwon Okubena, FCMA Health Forever International11 Dipeolu StreetOff Obafemi Awolowo WayIkeja, Lagos 100001NigeriaE-mail:  okubena@health-forever.com 4 AYUBA ET AL.
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