A randomized comparative clinical trial study to evaluate the efficacy of modified Mustadi Yapana basti and Bhadranimbadi Asthapana basti in the management of Prameha Poorvarupa w.s.r to prediabetes

Diabetes is spreading throughout the globe rapidly in the 21st century. At present worldwide population of 387 million (8.3% of world population) is diabetic which is predicted to reach 592 million by 2035 Evidence increasingly demonstrates that pre-diabetes is a toxic state that increases the risk for diabetes, cardiovascular disease, non-alcoholic fatty liver disease, neuropathy, chronic kidney disease, cancer and dementia, as well as all mortality due to various pathophysiological changes in several tissues and organs. Unfortunately, use of available evidence- based treatment for pre-diabetes is low. Pre-diabetes is a major health burden associated with an increased risk of subclinical metabolic disturbances and fulminant disease.The Indian diabetes program -1 (IDPP-1) has shown an annual incidence of approximately 18% among subjects with impaired glucose tolerance.There are an estimated 77.2 million people in India who are suffering from pre-diabetes. India has 10%-15% pre-diabetes prevalence with possibly world’s highest pre-diabetes into diabetes conversion rate (18%) prediabetic every year in India converts into overt diabetes Diabetes onset in India is nearly two decades earlier comparing to rest of the world. Prevalence rate of pre-diabetes in rural and urban Gujarat is 8.4% and 11.5% respectively. Hence, it is a public health menace causing morbidity, mortality and also impacting socio-economy of  the affected individual. So early detection and prevention of pre-diabetes is need of  the hour to prevent or delay diabetes and other microvascular and macrovascular disorders.                                                                                                                                                                   In additional health care system native to Indian subcontinent is found to have vivid clinical description of prameha/madhumeha, a condition mimicking diabetes. It has vast description of aetiology associated with diabetes and also has a description about its pre-clinical features(poorvarupa). Prameha is bahudoshajanya vyadhi and the main treatment principle in the condition of prediabetes is the elimination of kleda(wetness)shodhana chikitsa is the best choice of treatment for kledaharana Acharya Vagbhata has mentioned 5 types of shodhana treatment modalities which is known as panchshodhana and Niruha basti is one of them. Basti is consider to be the best treatment to normalize the vata dosha which is mainly involved in this conditions. Basti is the Ardhachikitsa among all therapeutic measures also it has vast field of action.As this condition is kapha pradhana vyadhi, the drugs of bhadranimbadi asthapana basti are ushna virya and kapha vata hara property. It regulates the samana vayu and restores the jatharagni to normal, also it activates the vyana vayu which break srotosanga and synergize the activity of basti at cellular level. Acharya Charaka mentioned mustadi yapana basti and the drugs used in mustadi yapana basti are tikta dravyas which are raktaprasadaka and therefore it also helps to break the avarodha due to kleda and improves blood sugar levels. This study investigates the effects of modifying both basti through the addition of takra. As takra acts as grahi, laghu, Deepana, also it is beneficial for kapha dosha due to its Kashaya rasa, ushna virya, vikasitva gunas and it aids in stimulating agni. In Ayurveda, panchkarma  plays an important role by restoring metabolic activities, eliminating toxins, strengthening tissue function and preventing re-occurrence of the disease.

 NEED OF STUDY:

In modern science, there isn’t a specific treatment for prediabetes, instead healthcare professionals recommend  lifestyle modifications such as dietary changes and increased physical activity to manage the condition. Many individuals diagnosed with prediabetes, despite being aware of the risks, struggle to implement necessary lifestyle modifications. The challenge often stems from entrenched habits, societal influences, and a lack of perceived urgency. Inertia sets in as people find it difficult to break free from familiar patterns. Procrastination and misconceptions about the severity of prediabetes can lead to complacency.

Limited research exists on prediabetes in Ayurveda, hindering comprehensive understanding and treatment approaches. Hence, it is the need of time to find out a safe and effective treatment for pre- diabetes and here comes the role of ayurveda. In the classical literature clinical descriptions of prameha/madhumeha, a condition closely resembling modern-day diabetes. Ancient texts provide extensive information on the various factors contributing to diabetes and even describe pre-clinical symptoms (poorvarupa). Prameha is recognized as Bahudoshajanya vyadhi.

A cornerstone of prediabetes management lies in eliminating kleda (wetness). Shodhana chikitsa, particularly Niruha basti, emerges as a prime treatment modality for kledaharana (reduction of wetness). Acharya Vagbhata outlines five primary shodhana treatment modalities, collectively known as panchshodhana. Among these, Niruha basti stands out as a particularly effective treatment for balancing vata dosha, which plays a crucial role in this condition.

Basti is revered as the "Ardhachikitsa," signifying its paramount importance among all therapeutic measures. This treatment modality exhibits a broad spectrum of therapeutic applications. In this study*,* modified Asthapana basti and yapana basti are to be administered along with some lifestyle modification for improving the quality of life                                                                                                                            Acharya Sushruta indicated Asthapana basti in prameha. ‘Asthapana’ indicates establishment of life span and age, it corrects deranged metabolism and removes excess of morbid doshas. Yapana basti is a special kind of basti which are having the property to support life and promote longetivity. Since this condition is kapha pradhana vyadhi, the drugs employed in bhadranimbadi asthapana basti possess ushna virya and exhibit kapha vata hara properties. This treatment modality regulates samana vayu, restores jatharagni to its normal state, and activates vyana vayu, which effectively breaks down srotosanga and synergistically enhances the cellular level activity of basti. Acharya Charaka has mentioned mustadi yapana basti, and the drugs utilized in this formulation are tikta dravyas, known for their raktaprasadaka properties. Consequently, mustadi yapana basti also contributes to breaking down the avarodha caused by kleda and facilitates an improvement in blood sugar levels. This study delves into the effects of modifying both basti procedures through the incorporation of takra. As takra exhibits grahi, laghu, and Deepana properties, and is also beneficial for kapha dosha due to its Kashaya rasa, ushna virya, and vikasitva gunas, it effectively aids in stimulating agni.

In ayurveda, most diseases are caused by an accumulation of ama. It can be formed as a result of reduced agni(Jatharagni and dhatvagni) this ama at a later stage of disease can lead to bodily tissues and can cause tissue disruption. This study focuses on observations of Dhatu shaithilyata due to disease & Ayu Sthapana by Asthapana Basti also, the efficacy of modified Bhadra nimbadi asthapana basti & modifiedMustadi yapana basti , on prameha poorvarupa as a supportive therapy to improve quality of life. According to bhavaprakash Nighantu takravarga, takra has been mentioned as takra aids in stimulating the agni, also as it is santarpanotthita vikara it helps in reducing kapha and pittawhile balancing vata.  The medicated takra used in both basti is probiotic in nature. The  Pilot study has been conducted on modified mustadi yapana basti andmodified bhadranimbadi asthapana basti have suggested highly significant reduction in Atiswedapravritti(60%), karapadadaha(75%), and in angasada(80%) also there was significant reduction occur in measurements of FBS and PPBS. Recent cumulative evidence collectively indicates that there is compositional shift of gut microbiome on concord with prediabetes.                                                                                                                                                                In prediabetes, there is lower diversity of butyrate producing bacteria. It improves the intestinal barrier integrity, thereby helps in preventing the progression of prediabetes to diabetes. This comparative clinical study will assess changes in specific gut bacterial populations before and after Basti Karma through stool culture tests, also pharmaceutical and analytical standardization of Modified Bhadra Nimbadi Asthapana Basti and modified mustadi yapana basti procedures.

v AIM:-

 Ø To Compare the Efficacy of Modified Mustadi Yapana basti and Modified Bhadranimbadi Asthapana Basti along with lifestyle Modification in the Management of Pre-diabetes.

                                                       v OBJECTIVES:-

 1.    To Compare the Efficacy of Modified Mustadi yapana basti and  Modified Bhadranimbadi Asthapana Basti in the management of Pre-diabetes.

2.    To assess the specific probiotic strains of lactobacillus in basti formulations through microbiological analysis.

3.    To Evaluate the role of Modified Mustadi Yapana basti and Modified Bhadranimbadi Asthapana Basti  in improving gut microbioma in pre-diabetes(through stool culture test)

4.    Standardization of modified Mustadi Yapana Basti and modified Bhadranimbadi Asthapana basti through analytical and pharmaceutical study.

HYPOTHESIS :

Null Hypothesis (HO)

Ø There is no significant effect of Modified Bhadranimbadi Asthapana Basti in management of Pre-diabetes in comparision with Modified Mustadi Yapana Basti.

Alternative Hypothesis(H1)

Ø Modified Bhadranimbadi Asthapana Basti exhibits significant effect in the management of pre-diabetes in comparison with Modified Mustadi Yapana Basti.

 Alternative Hypothesis(H2)

Ø Modified Bhadranimbadi Asthapana basti and Modified Mustadi Yapana Basti both are equally effective in the management of pre-diabetes.

Research Question:

Is Modified Mustadi Yapana  basti and Modified Bhadranimbadi Asthapana  basti effective in management of  Prediabetes and gut health?

Methodology:

Materials and Methods:-

**1.**Source of data

**A.**Literature source:

All the available literature on prameha poorvarupa with special reference to pre-diabetes in classical and modern literature will be collected, screened, reviewed and analyzed including articles from reputed scientific journals and all internet sources.

**B.**Pharmaceutical source:

Medicine will be prepared in the GMP Certified pharmacy and Modified bhadranimbadi asthapana basti and Modified mustadi yapana basti will be prepared as per proper SOP’s.

**C.**Clinical source:

OPD and IPD of Panchakarma Department

D. Instrumental source:

Ø Preparation of basti :

a) Khalva yantra

b)    Stainless steel container

c)    Gas stove

d)    Spatula

e)    Measuring cup

f)     filter

Ø Stool culture(Tools and equipments)

a)    Laminar flow

b)    Autoclave

c)    Incubator

d)    Colony counter

e)    Water still

f)     Dispensing bottles

g)    Mccartney bottles

h)    Fecal sample

i)     Culture media

j)     Water Blanks

k)    Sterile pipettes

l)     Sterile petridish

m)  Spatula

n)    Mark pens

o)    Dust coat, nose marks, and gloves

DRUG REVIEW

Modified Bhadranimbadi Asthapana Basti preparation and standardization involves following steps:

·      Bhadranimbadi Asthapana Basti: (Su.sa.chi 38/60-62)

·      (Ksheerpaka reference – Sharangdhara Samhita)

·      Takra as basti dravya: (Cha.vi.8/140 amalakaskanda)

Preparation of ksheerpaka

1.    The preparation of ksheerpaka using the kwatha dravyas like Bhadra, nimba, guduchi, sariva, and brhati about one part of each mentioned above drug will be added, eight parts of milk and thirty-two parts of water is to be added and heated till only the milk part remains. Moderate temperature will be maintained throughout the preparation of ksheerpaka.

2.    Preparation of dadhi from ksheerpaka

The mixture of ksheerpaka is filtered well, when the ksheerpaka is lukewarm little quantity of fresh curd is added and kept overnight. The medicated dadhi obtained is added with water and churned well to make it takra.

3.    Preparation of Basti

Sr.No.

Dravya

Quantity

|1

Madhu

80 ml

|2

Saindhava Lavana

12 gms

|3

Sneha-  Murchita tila tailam

60 ml

|4

Kalka- vaca, madanaphala, sunthi, bilva,

18 gms

|5

Bhadra, Nimba, Guduchi, Sariva, brhati

Processed into ksheerapaka & Takra

310 ml

| Total – Takra Basti

480 ml

Kalka will be prepared using fine powders of vaca, madanaphala, sunthi, bilva. The paste will be made by adding water.

Madhu and saindhava lavana will be mixed together and triturated in khalva yantra till it becomes homogenous followed by addition of Sneha, kalka, takra. Each drug will be added after mixing of previous drug. The mixture thus prepared acquires a physical state of emulsion.

At each step, relevant observation will be made such as rotation per minute, the change in colour of mixture after adding the successive ingredients. These parameters will be observed and documented at each step. The final product will assess the HPTLC/TLC, pharmaceutical and analytical tests

Modified Mustadi Yapana Basti Preparation and Standardization involves following steps:

 ·      Mustadi Yapana Basti: (Cha.si.12/15)

·      Takra as basti dravya: (Cha.vi.8/140 amlakaskanda)

·      Musta prayoga in santarpanotthita vikaras :(cha.su.23/12)

 Preparation of ksheerpaka

1.    The preparation of ksheerpaka using the kwatha dravyas like musta, aragvadha, haritaki, bibhitaki, rasna , guduchi, bruhati, Punarnava and gokshura about one part of each mentioned above drug will be added, eight parts of milk and thirty-two parts of water is to be added and heated till only the milk part remains. Moderate temperature will be maintained throughout the preparation of ksheerpaka.

2.    Preparation of dadhi from ksheerpaka

The mixture of ksheerpaka is filtered well, when the ksheerpaka is lukewarm little quantity of fresh curd is added and kept overnight. The medicated dadhi obtained is added with water and churned well to make it takra.

3.    Preparation of Basti

Sr. no

Dravya

Quantity

|1

Madhu

80 ml

|2

Saindhava

12 gms

|3

Murchita Tila Tailam

60 ml

|4

Shatapushpa Kalka

18 gms

|5

Musta, aragvadha, haritaki, bibhitaki, rasna, guduchi, Punarnava, brhati, gokshura processed into Ksheerapaka & Takra

310 ml

|            Total  - (Takra Basti)

480 ml

Kalka will be prepared using fine powders of shatapushpa. The paste will be made by adding water.

Madhu and saindhava lavana will be mixed together and triturated in khalva yantra till it becomes homogenous followed by addition of Sneha, kalka, takra. Each drug will be added after mixing of previous drug. The mixture thus prepared acquires a physical state of emulsion.

At each step, relevant observation will be made such as rotation per minute, the change in colour of mixture after adding the successive ingredients. These parameters will be observed and documented at each step. The final product will assess the HPTLC/TLC, pharmaceutical and analytical tests.

METHOD OF COLLECTION OF DATA:

a. SAMPLE SIZE : 20 :-

 Considering sample size of 20 patients in clinical trial which will randomize in two groups.

 GROUP A – 10 patients will be administered Modified mustadi yapana basti.

 GROUP B – 10 patients will be administered Modified bhadranimbadi asthapana basti.

  b. STUDY DESIGN : Randomized Comparative Clinical Trial Study.

  c. DURATION OF TREATMENT: 10 days for each group

d. STUDY DURATION: 18 months

e. FOLLOW UP: 16 days

DIAGNOSTIC CRITERIA:

According to WHO :

Ø FPGT : 110-125 mg/dl

Ø OGTT: 140 – 200 mg/dl

Ø HBA1C : 5.7% to 6.4% (acc. To ADA**)**

Ø Urine Sugar

Ø Urine Albumin

Ø BMI( 22.2 kg/m2- 29.9 kg/m2 ) (acc. to WHO)

·      Inclusion Criteria:

1.    Patients of either gender between age group of 18 to 60.

2.    Patients fulfilling the criteria of diagnosis according to American Diabetes Association(HbA1C – 5.7% to 6.4%)

*3.*Basti Arhas (patients suitable for basti karma)

4.    Patients having BMI 22.2 kg/m2- 29.9 kg/m2 (acc. to WHO)

5.    Patients willing to sign informed consent and able to participate in the study procedure.

 ·      Exclusion Criteria:

1.    K/C/O Diabetes Mellitus, Insulin Dependent Diabetes, Juvenile Diabetes Mellitus and Gestational Diabetes.

2.     K/C/O Major Systemic Diseases like IHD, Malignancies, Uncontrolled Hypertension.

3.    Patients on prolonged medication (>6weeks) with Corticosteroids or any other Drug that may have an influence on the outcome of the study.

4.    Pregnant and lactating women.

5.    Patients with active infective diseases like Tuberculosis, Hepatitis B.

6.    Immunocompromised patients.

*7.*Basti Anarha(Patients not suitable for basti karma)

        ·      ASSESSMENT CRITERIA:

 The patients will be examined before Basti karma and after completing the 8 days basti schedule. Changes in the patient’s condition will be noted and the following points will be taken into consideration for the assessment of the results.

The effect of the basti (Panchkarma therapy) will be assessed based on the following criteria:

ü Samyak Basti Lakshanas

ü The retention time of basti

ü Number of purisha vega

ü Possible complications after administration of basti

ü Vital data of patients

 ·      SUBJECTIVE CRITERIA:

1.    Angasada (Physical fatigability)

2.    *Sheeta Priyatvam(*Desire for cold food and environment)

3.    Atiswedapravritti (Excessive sweating)

4.    Karapadadaha (Burning sensation in hands and feet)

5.    Shithilangata (Flabbiness of body)

6.    Gala-talu shosha (Dryness of the throat and palate)

 Ø Gradation scales:(Ref- National institution of health)

ANGASADA

 

None(No
fatigue)

0

|Mild(occasionally)

1

|Moderate(Most time of day)

2

|Severe(Always)

3

  

SHEETA PRIYATVAM

 

No desire

0

|Mild(occasionally)

1

|Moderate(during hot climate/2-3 times of the day)

2

|Severe(All the time)

3

 

ATISWEDAPRAVRITTI

 

None(After heavy work or in hot
weather)

0

|Mild(Profuse sweating after moderate work)

1

|Moderate(Sweating after little work)

2

|Severe(Profuse sweating after little work)

3

 

KARAPADADAHA

 

None(No burning sensation on hands
and feet)

0

|Mild(occasionally)

1

|Moderate(Most time of the day)

2

|Severe(Always)

3

 

SHITHILANGATA

 

None

0

|Mild(Only in few areas)

1

|Moderate(flaccidity especially limbs)

2

|Severe(Major areas)

3

 

GALA-TALUSHOSHA

 

None(No dryness of the throat and
palate)

0

|Mild(occasionally)

1

|Moderate(Most time of the day)

2

|Severe(Always)

3

OBJECTIVE CRITERIA:

 •       FPGT : 110-125 mg/dl

•       OGTT: 140 – 200 mg/dl

•       HBA1C : 5.7% to 6.4% (acc. To ADA**)**

•       Urine routine/Micro

•       CBC

•       Stool culture( before and after basti treatment)

·      PHARMACEUTICAL AND ANALYTICAL STUDY FOR STANDARDIZATION:

 Ø Raw drugs – Identification and collection

Ø Assessment of genuinity of raw materials

Ø Preparation of Modified Mustadi yapana basti and modified Bhadranimbadi Asthapana basti

Ø Analytical study of three samples for its physico-chemical characteristics and HPTLC/TLC of both formulations.

 

1.
Modified Mustadi yapana basti

(SAMPLE A,B,C)

   Madhu, saindhava, Sneha – murchita tila taila, kalka- shatapushpa, ksheerpaka prepared with kwatha of musta, bibhitaki, haritaki, aragvadha,rasna,punarnava,guduchi,brhati,gokshura and is processed to takra preparation

|*2.*Modified Bhadranimbadi Asthapana Basti

(SAMPLE A,B,C)

     Madhu, saindhava, sneha – murchita tila taila kalka - vaca, madanaphala, sunthi, bilva, ksheerpaka prepared with kwatha of bhadra, nimba, guduchi, sariva, brhati, and is processed to takra preparation-

  1.    Organoleptic characters of samples

2.    Sp. Gravity

3.    Ph value

4.    Reletive Viscocity

5.    Refractive index

6.    Total solid content

7.    Ash value

8.    HPTLC/TLC

9.    Emulsion test:

TEST NAME

SAMPLE A

SAMPLE B

SAMPLE C

|Dilution

    |Conductivity test

    |Dye test

   ASSESSMENT OF CHANGES IN GUT BACTERIAL POPULATIONS BEFORE AND AFTER BASTI THERAPY:

STOOL CULTURE TEST:

1.    Collection of stool sample in clean disposable container

2.    Wash your hands with soap and running water

3.    Specific bacteria & media:

a.     G. Clostridium – BSM agar

b.    Faecalibacterium prausnitzii – Enterococcus selective agar

c.     Lactobacillus plantarum HACO1 – MRSA vancomycin agar

d.    Escherichia – Sorbitol macconkey medium

e.     Streptococcus – Blood agar media

4.    Preparation of media plates by autoclaving.

5.    Stool sample diluted to 10-5 and 10-6 using saline

6.    The same is inoculated on to plates and incubated

7.    Positive and negative control plates will be maintained at all steps

8.    Colonies of bacteria are counted

9.    The results would be noted as number of colony forming units (CFU) in each sample

To evaluate the number of each bacterial populations mentioned above ( before and after treatment)

 ENUMERATING BACTERIA

a) Serial Dilution

Usually, one gram of faecal can have millions of micro-organisms. This makes it hard to count the

colonies if they are grown without being diluted because they become too crowded on Petri dishes.

Serial dilution is therefore meant to reduce the concentration of microbes in solution for easier

counting and estimation of faecal bacteria.

b) Data Analysis and Reporting

• Determination of the number of bacterial cells in a faecal sample is done as in the

equations below:

No. of bacterial cells /1gm moist faecal = number of colonies × inverted dilution

Weight of dry faeces

 No. of bacterial cells /1gm dry faecal = number of colonies ×inverted dilution

Weight of dry faeces

• The unit of measurement here is colony forming units (CFUs)/ g of faeces, where the

colony may be the yields of the growth and multiplication of a single cell or more.

   INTERPRETATION:

 

BACTERIA

NORMAL

PRE-DIABETES

SIGNIFICANCE

|G. Clostridium

High

Low

It helps in reducing fasting glucose levels, improves insulin sensitivity, increases butyrate producing bacteria, reduced levels of it can lead to insulin resistance.

|Faecalibacterium prausnitzii

High

Low

It helps in producing butyrate and SCFA’S, supplies to gut epithelium, also helps to reduce inflammation and oxidative damage.

|Lactobacillus Plantarum HACO1

High

Low

It helps in reducing adipose tissue accumulation and improves glycemic profile

|Escherichia

Low

High

It can lead to oxidative stress and insulin resistance

|Streptococcus

Low

High

It can lead to insulin resistance.

    Methodology to assess the specific probiotic strains of lactobacillus in basti formulations through microbiological analysis:

Two samples will be collected

Sample 1 – Modified bhadranimbadi asthapana basti

Sample 2 – Modified mustadi yapana basti

Enumerating bacteria – lactobacillus strains( Lactobacillus leichmannii, lactobacillus casei, lactobacillus delbrueckii, lactobacillus brevis, lactobacillus fermentum, lactobacillus caogulans, lactobacillus acidophilus, lactobacillus lactis, lactobacillus rhamnosus)

The medium selected for lactobacillus strain was MRS vancomycin agar

A loopful of sample was streaked on sterile MRS agar petri plate by quadrant streaking method, under aseptic conditions

Then they were incubated at 370C for 24 to 48 hrs.

After incubation, colonies were restreaked on MRS agar petri plate for formation of isolated colonies.

   INTERVENTION SCHEDULE:

Ø Modified bhadranimbadi asthapana basti schedule is as follows:

METHOD OF COLLECTION OF DATA:

a. SAMPLE SIZE : 20 :-

 Considering sample size of 20 patients in clinical trial which will randomize in two groups.

 GROUP A – 10 patients will be administered Modified mustadi yapana basti.

 GROUP B – 10 patients will be administered Modified bhadranimbadi asthapana basti.

  b. STUDY DESIGN : Randomized Comparative Clinical Trial Study.

  c. DURATION OF TREATMENT: 10 days for each group

d. STUDY DURATION: 18 months

e. FOLLOW UP: 16 days

DIAGNOSTIC CRITERIA:

According to WHO :

Ø FPGT : 110-125 mg/dl

Ø OGTT: 140 – 200 mg/dl

Ø HBA1C : 5.7% to 6.4% (acc. To ADA**)**

Ø Urine Sugar

Ø Urine Albumin

Ø BMI( 22.2 kg/m2- 29.9 kg/m2 ) (acc. to WHO)

·      Inclusion Criteria:

1.    Patients of either gender between age group of 18 to 60.

2.    Patients fulfilling the criteria of diagnosis according to American Diabetes Association(HbA1C – 5.7% to 6.4%)

*3.*Basti Arhas (patients suitable for basti karma)

4.    Patients having BMI 22.2 kg/m2- 29.9 kg/m2 (acc. to WHO)

5.    Patients willing to sign informed consent and able to participate in the study procedure.

 ·      Exclusion Criteria:

1.    K/C/O Diabetes Mellitus, Insulin Dependent Diabetes, Juvenile Diabetes Mellitus and Gestational Diabetes.

2.     K/C/O Major Systemic Diseases like IHD, Malignancies, Uncontrolled Hypertension.

3.    Patients on prolonged medication (>6weeks) with Corticosteroids or any other Drug that may have an influence on the outcome of the study.

4.    Pregnant and lactating women.

5.    Patients with active infective diseases like Tuberculosis, Hepatitis B.

6.    Immunocompromised patients.

*7.*Basti Anarha(Patients not suitable for basti karma)

        ·      ASSESSMENT CRITERIA:

 The patients will be examined before Basti karma and after completing the 8 days basti schedule. Changes in the patient’s condition will be noted and the following points will be taken into consideration for the assessment of the results.

The effect of the basti (Panchkarma therapy) will be assessed based on the following criteria:

ü Samyak Basti Lakshanas

ü The retention time of basti

ü Number of purisha vega

ü Possible complications after administration of basti

ü Vital data of patients

 ·      SUBJECTIVE CRITERIA:

1.    Angasada (Physical fatigability)

2.    *Sheeta Priyatvam(*Desire for cold food and environment)

3.    Atiswedapravritti (Excessive sweating)

4.    Karapadadaha (Burning sensation in hands and feet)

5.    Shithilangata (Flabbiness of body)

6.    Gala-talu shosha (Dryness of the throat and palate)

 Ø Gradation scales:(Ref- National institution of health)

ANGASADA

 

None(No
fatigue)

0

|Mild(occasionally)

1

|Moderate(Most time of day)

2

|Severe(Always)

3

  

SHEETA PRIYATVAM

 

No desire

0

|Mild(occasionally)

1

|Moderate(during hot climate/2-3 times of the day)

2

|Severe(All the time)

3

 

ATISWEDAPRAVRITTI

 

None(After heavy work or in hot
weather)

0

|Mild(Profuse sweating after moderate work)

1

|Moderate(Sweating after little work)

2

|Severe(Profuse sweating after little work)

3

 

KARAPADADAHA

 

None(No burning sensation on hands
and feet)

0

|Mild(occasionally)

1

|Moderate(Most time of the day)

2

|Severe(Always)

3

 

SHITHILANGATA

 

None

0

|Mild(Only in few areas)

1

|Moderate(flaccidity especially limbs)

2

|Severe(Major areas)

3

 

GALA-TALUSHOSHA

 

None(No dryness of the throat and
palate)

0

|Mild(occasionally)

1

|Moderate(Most time of the day)

2

|Severe(Always)

3

OBJECTIVE CRITERIA:

 •       FPGT : 110-125 mg/dl

•       OGTT: 140 – 200 mg/dl

•       HBA1C : 5.7% to 6.4% (acc. To ADA**)**

•       Urine routine/Micro

•       CBC

•       Stool culture( before and after basti treatment)

·      PHARMACEUTICAL AND ANALYTICAL STUDY FOR STANDARDIZATION:

 Ø Raw drugs – Identification and collection

Ø Assessment of genuinity of raw materials

Ø Preparation of Modified Mustadi yapana basti and modified Bhadranimbadi Asthapana basti

Ø Analytical study of three samples for its physico-chemical characteristics and HPTLC/TLC of both formulations.

 

1.
Modified Mustadi yapana basti

(SAMPLE A,B,C)

   Madhu, saindhava, Sneha – murchita tila taila, kalka- shatapushpa, ksheerpaka prepared with kwatha of musta, bibhitaki, haritaki, aragvadha,rasna,punarnava,guduchi,brhati,gokshura and is processed to takra preparation

|*2.*Modified Bhadranimbadi Asthapana Basti

(SAMPLE A,B,C)

     Madhu, saindhava, sneha – murchita tila taila kalka - vaca, madanaphala, sunthi, bilva, ksheerpaka prepared with kwatha of bhadra, nimba, guduchi, sariva, brhati, and is processed to takra preparation-

  1.    Organoleptic characters of samples

2.    Sp. Gravity

3.    Ph value

4.    Reletive Viscocity

5.    Refractive index

6.    Total solid content

7.    Ash value

8.    HPTLC/TLC

9.    Emulsion test:

TEST NAME

SAMPLE A

SAMPLE B

SAMPLE C

|Dilution

    |Conductivity test

    |Dye test

   ASSESSMENT OF CHANGES IN GUT BACTERIAL POPULATIONS BEFORE AND AFTER BASTI THERAPY:

STOOL CULTURE TEST:

1.    Collection of stool sample in clean disposable container

2.    Wash your hands with soap and running water

3.    Specific bacteria & media:

a.     G. Clostridium – BSM agar

b.    Faecalibacterium prausnitzii – Enterococcus selective agar

c.     Lactobacillus plantarum HACO1 – MRSA vancomycin agar

d.    Escherichia – Sorbitol macconkey medium

e.     Streptococcus – Blood agar media

4.    Preparation of media plates by autoclaving.

5.    Stool sample diluted to 10-5 and 10-6 using saline

6.    The same is inoculated on to plates and incubated

7.    Positive and negative control plates will be maintained at all steps

8.    Colonies of bacteria are counted

9.    The results would be noted as number of colony forming units (CFU) in each sample

To evaluate the number of each bacterial populations mentioned above ( before and after treatment)

 ENUMERATING BACTERIA

a) Serial Dilution

Usually, one gram of faecal can have millions of micro-organisms. This makes it hard to count the

colonies if they are grown without being diluted because they become too crowded on Petri dishes.

Serial dilution is therefore meant to reduce the concentration of microbes in solution for easier

counting and estimation of faecal bacteria.

b) Data Analysis and Reporting

• Determination of the number of bacterial cells in a faecal sample is done as in the

equations below:

No. of bacterial cells /1gm moist faecal = number of colonies × inverted dilution

Weight of dry faeces

 No. of bacterial cells /1gm dry faecal = number of colonies ×inverted dilution

Weight of dry faeces

• The unit of measurement here is colony forming units (CFUs)/ g of faeces, where the

colony may be the yields of the growth and multiplication of a single cell or more.

   INTERPRETATION:

 

BACTERIA

NORMAL

PRE-DIABETES

SIGNIFICANCE

|G. Clostridium

High

Low

It helps in reducing fasting glucose levels, improves insulin sensitivity, increases butyrate producing bacteria, reduced levels of it can lead to insulin resistance.

|Faecalibacterium prausnitzii

High

Low

It helps in producing butyrate and SCFA’S, supplies to gut epithelium, also helps to reduce inflammation and oxidative damage.

|Lactobacillus Plantarum HACO1

High

Low

It helps in reducing adipose tissue accumulation and improves glycemic profile

|Escherichia

Low

High

It can lead to oxidative stress and insulin resistance

|Streptococcus

Low

High

It can lead to insulin resistance.

    Methodology to assess the specific probiotic strains of lactobacillus in basti formulations through microbiological analysis:

Two samples will be collected

Sample 1 – Modified bhadranimbadi asthapana basti

Sample 2 – Modified mustadi yapana basti

Enumerating bacteria – lactobacillus strains( Lactobacillus leichmannii, lactobacillus casei, lactobacillus delbrueckii, lactobacillus brevis, lactobacillus fermentum, lactobacillus caogulans, lactobacillus acidophilus, lactobacillus lactis, lactobacillus rhamnosus)

The medium selected for lactobacillus strain was MRS vancomycin agar

A loopful of sample was streaked on sterile MRS agar petri plate by quadrant streaking method, under aseptic conditions

Then they were incubated at 370C for 24 to 48 hrs.

After incubation, colonies were restreaked on MRS agar petri plate for formation of isolated colonies.

   INTERVENTION SCHEDULE:

Ø Modified bhadranimbadi asthapana basti schedule is as follows:

     

1st
day

2nd day

3rd day

4th day

5th day

6th day

7th day

8th day

|BAB

BAB

BAB

BAB

BAB

BAB

BAB

BAB

  BAB * -  Modified bhadranimbadi asthapana basti

 Ø Modified mustadi yapana basti schedule is as follows:

1st
day

2nd day

3rd day

4th day

5th day

6th day

7th day

8th day

|MYB

MYB

MYB

MYB

MYB

MYB

MYB

MYB

 MYB* - Modified mustadi yapana basti

 

Sr.
No.

Basti

Quantity

|1

Modified bhadranimbadi asthapana basti

480 ml

|2

Modified mustadi yapana basti

480 ml

 20 patients of pre-diabetes who fulfils the inclusion criteria will be randomly selected and assigned into 2 groups each comprising of 10 patients.

  STANDARD OPERATIVE PROCEDURE:

POORVAKARMA

PRADHANAKARMA

PASCHATKARMA

|BAB- Selection of the patient, counselling of the patient, basti preparation.

Deepana-pachana with chitrakadi vati 2-2-2 A/f

Sadhyo virechanafor koshtha shuddhi with eranda taila(10 ml)

 MYB – Selection of the patient, counselling of the patient, basti preparation.

Deepana-pachana with chitrakadi vati 2-2-2 A/f

Sadhyo virechana with eranda taila(10 ml)

  BAB – Modified Bhadranimbadi Asthapana Basti administration at Nati-kshudharta state(in the early morning, empty stomach)

MYB – Modified Mustadi Yapana Basti administration at Nati-kshudharta state (in early morning, empty stomach.)

BAB – Rest, snana, laghu bhojana advised, basti pratyagamana kala and number of vega will be noted.

MYB – Rest, snana, laghu bhojana advised*, basti pratyagamana kala* and number of vega will be noted.

DIET ADVISED: (12)

At
Wake up time :

One glass of warm water infused with one teaspoon of overnight-soaked fenugreek seeds

One cup sugar-free green or black tea

|Breakfast

One bowl of vegetables and muesli, or vegetable dalia (broken wheat porridge)

One scrambled egg white or boiled egg

One tiny apple or some papaya slices

|Mid- morning snack

A single cup of buttermilk

A small handful of nuts (walnuts or almonds)

|Lunch

One small bowl of brown rice or one or two whole wheat chapatis

One bowl of curry with mixed vegetables that uses little oil

One bowl of dal (lentil soup)

Cucumber, tomatoes, carrots, and lemon dressing on a side salad plate

One little bowl of plain yogurt

|Post lunch

One tiny bowl of roasted chana(chickpeas) or sprout salad

One cup sugar-free herbal or green tea

|Evening snacks

One tiny bowl of roasted makhana (fox nuts) or an orange or guava fruit

|Dinner

One small bowl of quinoa or two whole wheat chapatis One bowl of steamed or grilled veggies

One bowl of curry made of chickpeas or a piece of grilled chicken or fish

A tiny bowl of mixed salad greens

|Bed

• time

One cup of warm milk, ideally low-fat,

sprinkled with turmeric.