The Role of Vitamin D, A, and Beta Carotene in Tuberculosis Patients With Vitamin D Receptor Gene Polymorphism
Phase: NA
Conditions: Tuberculosis
Sponsor: Universitas Sumatera Utara

1 
A Randomized Control Trial: The Role of Vitamin D, A, and Beta 
Carotene in Tuberculosis Patients with Vitamin D Receptor Gene 
Polymorphism 
 
Dina Keumala Sari1*, Ridha Dharmajaya2, Mutiara Indah Sari3, Dewi Masyithah4 
1Tropical Medicine Study Program, Faculty of Medicine, Universitas Sumatera Utara 
2Neurosurgery, Department, Faculty of Medicine, Universitas Sumatera Utara 
3Biochemistry, Department, Faculty of Medicine, Universitas Sumatera Utara 
4Parasitology, Department, Faculty of Medicine, Universitas Sumatera Utara 
*Correspondence: dina@usu.ac.id; Tel.: +62-813-971-77-693 
 
Keywords. 25(OH)D serum, calcium, counseling, FokI, retinol, TaqI 
Abstract. Recent studies showed that vitamin D and A has an effect in improving sputum conversion in 
tuberculosis. This study aims to find out the effect of vitamin D 1000 IU and A 6000 IU supplementation on 
Tuberculosis patients with vitamin D receptor gene polymorphism, who live in North Sumatera, Indonesia. This 
study is a randomized control clinical trial, with 48 tuberculosis patients with vitamin D receptor gene 
polymorphism which are TaqI and FokI participating, divided into two groups, each with 24 participants, which 
are treatment group (I) which receives nutritional counseling, vitamin D 1000 IU, vitamin A 6000 IU, and 
control group (C) which only receives nutritional counseling for 28 days. Patients who participated was found to 
be heterozygous with TaqI (T>C) or FokI (C>T) genotype variants. The result of this study showed that at the 
start, serum 25(OH)D levels in group I were lower compared to group C (19.746.59 ng/mL vs 25.217.57 
ng/mL). Group I showed significant correlation between vitamin D level categories with sputum conversion 
(mean: standard deviation= 2.250.68 weeks). Supplementation of vitamin D 1000 IU provides an accelerated 
sputum conversion in tuberculosis patients with vitamin D receptor gene polymorphism. 
1 
Introduction  
Prevalence of tuberculosis (TB) is rising each year and is one of the causes of mortality in 
Indonesia and is also becoming a health problem in the world. Governments of various 
countries with the highest prevalence have also performed initiation and implementation of 
DOTS (directly observed treatment, short course) as well as possible and Stop TB Strategy all 
over the world, but new cases always appear and relapse cases because of resistance emerge. 1 
This condition is worsened by susceptibility from Mycobacterium tuberculosis infection. This 
susceptibility is caused by environmental, nutritional, and genetical factors. 2-4 Based on 
previous studies, it’s stated that existence of vitamin D receptor gene polymorphism has a 
firm influence on the high risk of tuberculosis disease. 2 
Vitamin D receptor gene polymorphism is related to vitamin D deficiency and is related to 
an increase risk of tuberculosis disease. Several of the related genes are FokI and TaqI which 
are related to high vitamin D deficiency and in the end is related to the role of immune 
regulation in the body.2, 4, 5 Vitamin D supports the induction of pleiotropic antimicrobial 
response in tuberculosis patients. 6, 7 Supplementation of vitamin D may accelerate sputum 
conversion and support improvement of tuberculosis patients’ clinical condition and the 
mechanism of action is through an increase in immune cells by activating 25-hydroxyvitamin 
D receptor.7, 8 Vitamin D does not work alone, it performs interaction with other vitamins or 
minerals 9, 10. There is a decrease of several vitamins, for example vitamin A and carotenoid 

2 
which is caused by low nutritional intake because of loss of appetite and lost from urine 
secretion because of severe infection 10, 11. 
The role of vitamin A and beta carotene also affects the improvement of tuberculosis 
patients’ clinical condition 9, 10, 12. Previous studies showed that interaction between vitamin A 
and D in improving tuberculosis patients’ clinical condition, low level of vitamin A, which is 
retinol, is affected by acute phase response (APR). During that APR process, processes of 
transcription and translation of positive acute phase response proteins (such as C-reactive 
protein/CRP) experience an increase 1, 13, 14. While the process occurs, the role of protein is 
highly necessary, which is retinol binding protein, with the role as the primary transport 
protein to move vitamin A from liver storage towards peripheral tissues and during the APR 
process 9. Based on this, then the role of protein in the improvement process of tuberculosis 
patients’ clinical condition is highly necessary, tuberculosis patients’ malnutrition condition 
with low level of protein in the body 7, 15. 
Vitamin D deficiency is found in tropical areas like Indonesia, it even occurs in healthy 
individuals. It’s predicted that lack of vitamin D does not occur because there is enough 
amount of sun light, although aside from sun-evading lifestyle, in addition to lack of vitamin 
D intake, then the prevalence of vitamin D deficiency is increasing, although several previous 
studies stated that vitamin D levels do not affect calcium levels. Calcium has a role in 
regulation of tuberculosis infection through its role in cellular activity in tuberculosis 
infection. 
Vitamin D’s mechanism of action is among others by modulating the body’s immune 
response starting from the increase of vitamin D receptor (VDR) activity, and after that it will 
activate monocytes as the body’s immune cell 7, 16, 17. Another mechanism is the active form 
of vitamin D which is 1,25(OH)2D3 will hinder the proliferation of T cell and increase the 
abilities of pathogen-activated macrophages 15, 17. 
Vitamin D receptor is a member of the nuclear receptor of transcription factors, with 
activation of 1.25(OH)2D3, binding DNA response element and form heterodimer with 
retinoic acid receptor (RXR). This heterodimer will link the relation between vitamin D and 
vitamin A or carotenoid, including beta carotene. 
Because of the big role of both vitamins and carotenoid, a clinical randomized trial will be 
performed with supplementation of vitamin D 1000 IU and vitamin A 6000 IU for 28 days on 
new cases of tuberculosis patients in Public Health Center (Pusat Kesehatan Masyarakat) as 
the referral center of tuberculosis cases in the City of Medan. Tested parameters are levels of 
25(OH)D, retinol, beta carotene, and calcium before and after treatment. 
2 
Methods 
The aim of this study is to find the effect of vitamin D 1000 IU and vitamin A 6000 IU 
supplementation along with nutritional counseling on sputum conversion, for 28 days on new 
cases of tuberculosis patients who visit three Public Health Centers, which are Teladan Public 
Health Center, Amplas Public Health Center, and Belawan Public Health Center, which are 
located in the City of Medan, North Sumatera, Indonesia. The location of sample collection is 
in Sumatera Island with latitude 3.57 N and longitude 98.65 E, with enough amount of sun 
light, temperature of ± 32 oC (90 oF). The study was performed during the period of May to 
September 2019. This study is a controlled randomized clinical trial, after going through 
inclusion and exclusion criteria, 48 patients participated in the study, divided into 24 patients 
in the treatment group (I) and the control group (C) has 24 patients (Figure 1).  
Research subjects were males or females, new cases tuberculosis patients with AFB (+) 
who fulfills inclusion and exclusion criteria, chosen through purposive sampling technique. 
The inclusion criteria are new case lung tuberculosis patient with acid fast bacilli (AFB) result 

3 
(+), aged 18-60 years, heterozygote genotype test on one of the genes TaqI and FokI, willing 
to participate in the study and complete the informed consent. The exclusion criteria are 
pregnant, found to have comorbidities such as diabetes mellitus or Human Immunodeficiency 
Virus (+), and Tuberculosis on category II. Research subjects will drop out if they don’t 
consume both supplements for more than 14 days and if they experience worsening clinical 
condition, although they will still be included when blood samples were drawn at the end of 
the study. 
Randomization process 
Sample sized determined by using formula based on tuberculosis prevalence in Medan City. 
In order to avoid bias in the study, this was double blind trial. Randomization was performed 
through block randomization. This action was also taken in order to remove selection bias, 
names of research subjects were assigned codes, and later a block randomization process was 
performed by using block table and divided by using closed envelope in order to divide 
treatment and control groups.  
Treatment 
The treatment group were given vitamin D 1000 IU supplementation in the form of soft gel 
and vitamin A 6000 IU in the form of tablet, along with nutritional counseling. Meanwhile, 
the control group only received nutritional counseling, but no supplementation. 
Supplementation were given once a day for 28 days, monitoring and administration of 
supplementation were given every two weeks in order to inquire adherence towards 
supplementation consumption and perceived side effects. All research subjects went through a 
clearance period of one week and were requested not to consume any supplements. 
Afterwards, measurements were taken before and after treatment, covering four tests, which 
are for 25(OH)D, calcium, retinol level, and beta carotene serum level. 
Laboratory examinations for vitamin D, calcium, vitamin A, and beta carotene 
At the end of the study, analysis was performed on the whole sample data, storage of samples 
was done at a temperature of -80oC beforehand. Laboratory examiners did not know which 
specimens were case or control samples, samples were assigned codes and they were given 
randomly. Measurement of 25(OH)D serum level by using chemiluminescent immunoassay 
(CLIA) technology (Diasorin, Stillwater, MN) device, the measurement range was 4.0 and 
150 ng/mL. The lowest level was 4.0 ng/mL based on inter assay precision of 3.90% CV. 
Range of categories of vitamin D levels are <20 ng/mL considered deficiency, 20-30ng/mL 
(insufficiency), 30-100 ng/mL (sufficiency).18 Serum calcium level was measured by using 
ADVIA Bayer Assayed Chemistry Controls with reaction measurement 545/658 nm, and 
normal calcium serum level was 8.3-10.6mg/dL. Measurement of retinol and -carotene 
serum levels in this study was performed by using high-performance liquid chromatography 
(HPLC) device, with interassay coefficient variation for retinol and beta carotene was <5%. 
The normal range of retinol level was >0.7mol/L, while the normal range for beta carotene 
serum level was 0.3–0.6 mol/L.  
Patient sputum tests were performed in the laboratories of Teladan, Amplas, and Belawan 
Public Health Centers by using the Ziehl Neelsen method. Sputum collection was performed 
using the SPS (Sewaktu-Pagi-Sewaktu/Anytime-Morning-Anytime) 3 times method. 
Administered Anti Tuberculosis medications were Rifampicin, INH, Pyrazinamide, 
Streptomycin, Ethambutol with dosages adjusted to the patients’ body weight. AFB tests were 
performed before treatment, on week 2, 4, 6 and week 8 during the intensive phase treatment, 
afterwards data was collected and noted.  

4 
Continuous variables were expressed as continuous variables using means±SDs. In order to 
find differences of vitamin D, retinol, beta carotene, and calcium levels between the two 
groups before and after the treatment, analysis was performed by using the t test independent. 
Categorical variables were expressed as percentage proportions and in order to show the 
relationships between vitamin D deficiency and sputum conversion, chi-square test was used, 
and Fischer test if the data did not meet the criteria. The p values <0.05 were considered 
statistically significant. We used SPSS program (version 11.5; SPSS Inc, Chicago, IL) to 
perform the analysis. This study was carried out after ethical approval was obtained from the 
Health Research Ethics. Committee of Sumatera Utara University Medical School (No. 
135/TGL/KEPK FK USU-RSUP HAM/2019) and all participants were given written 
informed consent to the study procedures. 
3 
Results and Discussions 
This study has 48 research subjects taking part, after completing the screening process, 
through inclusion and exclusion criteria (Figure 1), from a total of 63 research subjects. After 
signing informed consent and going through block randomization, they were divided into two 
groups, which are the treatment and control groups each with 24 research subjects. During the 
study period, two research subjects couldn’t participate until the end of the period because 
they experienced nausea and didn’t routinely consume the supplements, although both 
research subjects participated in having their blood drawn at the end of the study as part of the 
research (intention to treat). 
Baseline characteristic 
Baseline characteristic of all research subjects showed that most of the tuberculosis 
prevalence was experienced by the Batak Tribe (this is probably because the location of the 
study is dominated by this tribe), married, male, younger age group, and low body weight 
(Table 1). Statistic testing did not show any difference between the groups, did not indicate 
any significant difference between the treatment and the control groups. For baseline 
characteristic, both groups are homogenous in nature. 
Polymorphism of vitamin D receptor gene influences 25(OH)D serum level in the body. 
Two widely studied vitamin D receptor genes were TaqI and FokI, especially in relation to 
tuberculosis. This study included vitamin D receptor gene polymorphism for all research 
subjects, with heterozygote genotype on vitamin D receptor gene which are TaqI (rs731236) 
or FokI (rs10735810). Hopefully, with vitamin D and A supplementation, both vitamins 
would synergize in improving clinical condition of tuberculosis patients aside from providing 
routine anti-tuberculosis regimens. In previous studies, FokI receptor gene polymorphism was 
related to increased risk of tuberculosis 2, 4, 5, 19-21. This study showed that patients with VDR 
gene polymorphism would exhibit vitamin D deficiency and insufficiency. Administration of 
vitamin D supplements showed an increase in vitamin D level, but it still did not show 
significant differences of 25(OH)D serum levels in both groups. The differences in vitamin D 
levels in both groups were significant, therefore administration of vitamin D supplementation 
was highly necessary in tuberculosis patients with VDR gene polymorphism. At the end of 
this study, significant result in sputum conversion was shown.  
Categories and laboratory parameter levels 
Table 2 shows that before the study, the percentage of vitamin D deficiency and insufficiency 
were found in higher numbers from all research subjects. None of the research subjects was 
categorized as normal, only reaching the sufficient category. Although, when compared to the 

5 
parameter categories from before the study, then vitamin D and calcium exhibited 
homogenous data. Meanwhile, study results for vitamin A and beta carotene categories 
showed significant differences between both groups for vitamin A (p=0.001) and beta 
carotene (p=0.001) before the start of the study. These results showed that from the laboratory 
test category before the study, vitamin D and calcium indicated no difference in both groups 
categories, but for vitamin A and beta carotene categories, there were differences. 
Other factors were body fat mass and profession. In this study, the body fat mass of 
tuberculosis patients was mostly low-fat level. Although high level of fat was still found, the 
percentage of vitamin D level categories were still considered as vitamin D deficiency and 
insufficiency. Low vitamin D levels at the start of the study was probably caused by the 
vitamin D receptor gene polymorphism, aside from other possibilities such as disease 
progressivity, sun-evading lifestyle, and lack of high vitamin D and vitamin A nutrition intake 
9, 22, 23. Types of profession also affected vitamin D levels. Jobs in which the person is mostly 
protected from sunlight such as indoor jobs would cause more pronounced vitamin D 
deficiency 24-27. 
The roles of vitamin A and beta carotene towards vitamin D activities hopefully would 
help with clinical improvement of tuberculosis patients 13, 14. The mechanism of action of 
vitamin A and beta carotene were through the role of retinoic acid, in which retinoic acid 
would be converted into 9-cis retinoic acid and into ligand which could activate RXR. RXR 
would then interact with VDR. That heterodimer would change protein conformation so that it 
may activate or repress molecules through gene transcription 9.  
The dimer forms of VDR-RXR would then bind on a specific sequence in the target 
promoter region called vitamin D response elements (VDRE). Several genes involved in the 
regulation of calcium, phosphor homeostasis, and vitamin D metabolism were related to 
VDRE. This transcription process would affect enzyme production involved in vitamin D 
metabolism such as enzymes involved in vitamin D synthesis. Increased vitamin D level in 
circulation would affect VDR activities in T lymphocytes, macrophages, and thymus tissues.  
Table 2 showed that after treatment, there was also no categorical difference in all groups, 
but not for laboratory parameter levels, which are for vitamin D, calcium, vitamin A, and beta 
carotene levels. This indicated that there was an increase in laboratory parameter levels 
though still not reaching normal values (Table 3). Table 3 showed that before the treatment, 
the data was inhomogeneous, which are for 25(OH)D serum level and beta carotene level, 
there were significant differences between both groups. In 25(OH)D serum level, it was seen 
that average 25(OH)D serum level was very low in the treatment group, while in the control 
group the average was high. Table 4 showed significant difference for the delta or difference 
of 25(OH)D serum levels during treatment. This indicated that with supplementation of 
vitamin D 1000 IU showed a significant increase in the treatment group compared to the 
control group, although there were other factors which made the levels unable to reach normal 
vitamin D category. This is probably caused by vitamin D receptor gene polymorphism. 
In this study, the low retinol levels found did not make way for a change in retinol serum 
levels compared to vitamin D levels after treatment. This was probably caused by the 
preparation of vitamin A supplements provided for the treatment group. Vitamin D 
administration in gel preparation provided good absorption. Bioavailability of the 
supplement’s preparation showed an increase of 25(OH)D serum levels, but the preparation of 
vitamin A supplement as tablets did not exhibit optimum bioavailability. The result of this 
study indicated that there was no significant increase in serum retinol. Whether or not this was 
caused by the tablet preparation or lack of food sources of vitamin A intake, which mostly 
came from animal produce such as liver and fish oil not being a frequently consumed food 
products in this study.  

6 
In this study, nutritional counseling was also performed, in which the benefits of vitamin A 
and provitamin A were also explained. Therefore, intake of food sources of provitamin A 
which contained a lot of carotenoid was increased. Significant improvement was shown on 
beta carotene levels in group C, although they didn’t receive beta carotene supplements, beta 
carotene intake probably may have been gained from easily acquired and affordable food 
sources. Food sources of beta carotene such as carrots, sweet potatoes, mango, sources from 
yellow-orange colored fruits, and dark green colored vegetables. This study did not find a 
significant difference between both groups, but it was shown through the increased levels, 
there was possibly a mechanism in which beta carotene held a role in sputum improvement. 
In Table 2, vitamin A categories showed significant differences between both groups 
before the study, but vitamin A levels did not exhibit differences before and after the study on 
both groups (Table 3). During the study, vitamin A levels did not show significant differences 
between both groups, this indicated that vitamin A supplementation did not show a difference 
on both groups (Table 4). This also applied for calcium and beta carotene levels which did 
not have significant differences on both parameters (Table 3 and 4). Beta carotene categories 
before the study showed significant differences on both groups, although not showing 
significant differences during treatment between both groups. 
Relationship of vitamin D status and sputum conversion 
Using chi-square analysis, conclusion was drawn that there was significant relationship 
between vitamin D categories and the week of sputum conversion (Table 5). It can be 
concluded that the fastest sputum conversion rely on the highest vitamin D level in the blood.  
Provision of counseling regarding increase of vitamin D food sources was also given. 
Although, vitamin D food sources belonged to expensive food products, such as codfish oil, 
salmon, or mushrooms. Fortification of vitamin D in milk is a good choice in fulfilling 
vitamin D intake. Sunlight is a renewable source in the tropical area, although jobs in which 
the workers are mostly covered from sunlight might make tuberculosis patients could not 
acquire adequate sunlight exposure in order to activate vitamin D from below the skin. 
Therefore, vitamin D supplementation in tuberculosis patients may provide more optimum 
synergistic effects by increasing beta carotene food sources18, 28-30.  
This study did not show any relations between calcium and vitamin D. Calcium intake also 
did not exhibit an increase in this study. Dietary sources of calcium such as milk, sour milk, 
or cheese are food products not commonly a part of daily meals for tuberculosis patients. This 
lack of calcium may be caused by the lack of consumption of such products by tuberculosis 
patients. This may be caused by high prices of those food or milk not being a commonly 
consumed food product. Other forms of calcium food sources are fish consumed along with 
their bones, such as anchovies31, 32. This form is a kind of food product which may be 
provided to tuberculosis patients in order to fulfill calcium requirements. 
This study did not show low serum calcium levels in the blood, even though the mean of 
serum 25(OH)D serum levels were low. In tuberculosis patients, there were found low levels 
of 25(OH)D, which should have an interaction between vitamin D and calcium. If the level of 
vitamin D in the blood is high, calcium absorption will increase. However, this mechanism 
did not occur in this study. Although 1000 IU of vitamin D had given per day for 28 days, and 
there had been an increased in vitamin D levels, but it has not reached normal range, and this 
did not affect calcium levels in the blood. Calcium which has a vital role in bone remodeling, 
support by vitamin D, which converted to calcitriol, would help the absorption of calcium in 
the intestine.  
About 300-500 mg of calcium derived from extracellular calcium as much as 900 mg, 
meaning that in the process of calcium, especially for bone remodeling, required 300-500 mg 
calcium. This amount will add to calcium intake from food sources, so it ranges from 1000-

7 
1500 mg33-35. However, this condition put the serum calcium in a homeostatic (balanced) 
state. Phosphorus also plays a role in the balance of calcium levels in the blood and the rate of 
calcium storage in the intestine. The process of calcium absorption with the support of 
vitamin D is significant for the metabolism of calcium and phosphorus36. Calcium absorption 
by the digestive tract will be increased, calcium and phosphorus from the bone mobilized, 
vitamin D controls expenditure and balance of minerals in the blood35. Although the calcium 
and vitamin D metabolism appeared to be altered in tuberculosis, there was no association 
between vitamin D and calcium categorization in this study. Previous study confirmed that 
serum calcium is raised in tuberculosis but the effect may be reduced by a low calcium 
intake37. 
However, with the length of this study, administration of vitamin D along with nutritional 
counseling, especially high intake of beta carotene food would show improved sputum 
conversion in the treatment and control group. Significant increase in vitamin D levels, 
although not exhibiting a difference between treatment and control group at the end of this 
study, showed clinical improvement of tuberculosis patients.  
This study has several limitations, which are not measuring parathyroid hormone levels 
and other clinical parameters of tuberculosis improvement, such as chest X-ray and blood 
inflammation parameters in the blood. 
4 
CONCLUSION 
Based on the result of this study, it can be concluded that tuberculosis patients with vitamin D 
receptor gene polymorphism (FokI and TaqI), may experience an increase in vitamin D level 
from supplementation of vitamin D 1000 IU and vitamin A 6000 IU along with nutritional 
counseling, which affected the acceleration of sputum conversion. 
ACKNOWLEDGMENTS 
The authors gratefully acknowledge that the present research is supported by Ministry of 
Research and Technology and the Higher Education Republic of Indonesia, Research and 
Community Service, Universitas Sumatera Utara. The support is under the research grant 
DRPM KEMENRISTEKDIKTI of the Year 2019 Contract Number: 11/UN5.2.3.1/PPM/KP-
DRPM/2019. 
The authors would also like to thank all research subjects, Head of the Health Office of the 
City of Medan, Heads of the Public Health Centers of Teladan, Amplas, Belawan, medical-
paramedical team, and students who have supported this research. 
COMPETING INTERESTS 
There are no funding or other conflicts of interest to declare for this research 
 
 
 

8 
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Indian J Clin Biochem 2014;29(3):377-81. 
 
 

10 
Table 1. Demographical characteristic description of all research subjects before treatment (n=48) 
Variable 
n 
Percentage 
Tribe 
Batak 
Javanese 
Mandailing 
Malay 
Other (Acehnese, Minang) 
 
28 
12 
2 
2 
4 
 
58.3 
25.0 
4.2 
4.2 
8.4 
Marita status 
Single 
Married 
 
17 
31 
 
 
35.4 
64.6 
Sex 
Male 
Female 
 
37 
11 
 
77.1 
22.9 
Age Categories (years) 
20-30 
30-40 
40-50 
50-60 
>60 
 
19 
16 
5 
6 
2 
 
39.6 
33.3 
10.4 
12.5 
4.2 
Body mass index 
Underweight  
Normal 
Overweight 
Obese 
 
16 
8 
15 
9 
 
33.4 
16.7 
31.3 
18.8 
 
 

11 
Table 2. Categories of vitamin D, calcium, vitamin A, and beta carotene levels of all research subjects 
before and after the study (n=48) 
Categories 
n 
Percentage 
Categories of vitamin D levels 
Before 
Deficiency   (<20) 
Insufficiency (20-32) 
Sufficiency   (32-54) 
After 
Deficiency   (<20) 
Insufficiency (20-32) 
Sufficiency   (32-54) 
 
Categories of Calcium 
Before 
Low 
Normal 
After 
Low 
Normal 
 
Categories of Vitamin A 
Before 
Low 
Normal  
After 
Low 
Normal  
 
Categories of Beta Carotene 
Before 
Low 
Normal 
High  
After 
Low 
Normal 
High 
 
 
19 
23 
6 
 
7 
29 
12 
 
 
 
9 
39 
 
11 
37 
 
 
 
13 
35 
 
8 
40 
 
 
 
6 
9 
33 
 
1 
6 
41 
 
 
39.6 
47.9 
12.5 
 
14.6 
60.4 
25.0 
 
 
 
18.8 
81.3 
 
22.9 
77.1 
 
 
 
27.1 
72.9 
 
16.7 
83.3 
 
 
 
12.5 
18.8 
68.8 
 
2.1 
12.5 
85.4 
 
 

12 
Table 3. Differences of vitamin D, calcium, vitamin A, and beta carotene levels in both groups before 
and after the study 
Parameter  
I  
(n=24) 
C  
(n=24) 
p 
Before 
Vitamin D 
Calcium 
Vitamin A  
Beta Carotene 
 
19.75 ± 6.59 
9.05 ± 0.51 
1.29 ± 0.81 
1.20 ± 1.27 
 
25.20 ± 7.57 
9.18 ± 0.49 
1.44 ± 0.69 
3.67 ± 2.06 
 
0.01 
0.35 
0.49 
0.001 
After 
Vitamin D 
Calcium 
Vitamin A 
Beta Carotene 
 
25.91 ± 4.96 
8.87 ± 0.54 
1.53 ± 0.77 
1.47 ± 1.22 
 
27.52 ± 6.44 
9.26 ± 0.56 
1.59 ± 0.75 
4.04 ± 2.04 
 
0.34 
0.17 
0.78 
0.001 
Note: data presented in the form of meanstandard deviation 
Analysis of differences between groups using t independent test. 
 

13 
 
Table 4. Change of vitamin D, calcium, vitamin A, and beta carotene levels in both groups 
Delta during the 
study 
I 
(n=24) 
C 
(n=24) 
p 
Delta Vitamin D 
Delta Calcium 
Delta Vitamin A 
Delta Beta Carotene 
6.16 ± 3.27 
-0.07 ± 0.44 
0.24 ± 0.15 
0.26± 0.22 
2.31 ± 2.62 
0.07 ± 0.23 
0.15 ± 0.16 
0.36 ± 0.23 
0.001 
0.14 
0.55 
0.16 
Data shown are means ± standard deviation 
1Analysed using independent t-test 
 

14 
Table 5. Relationship of vitamin D status and sputum conversion in the treatment group 
Vitamin D category 
Sputum conversion 
p value 
Second n (%) 
Fourth n (%) 
      Deficiency  
      Insufficiency 
      Sufficiency 
2 (8.3) 
9 (37.5) 
10 (41.7) 
 
2 (8.3) 
1 (4.2) 
0 (0) 
 
0.03 
Note: analyzed using chi square test