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International Journal of Pharmacy and Biological Sciences- ISSN: 2321-3272 (Print) IJPBS | Volume 3 | Issue 2 | APR – JUN | 2013 | 01-05 Research Article – Biological Sciences

PHYTOCHEMICAL ANALYSES OF ETHANOL AND WATER EXTRACTS OF MILLETTIA ABOENSIS, CUSCUTA REFLEXA, DANIELLA OLIVERI AND SYNCLISIA SCABRIDA Adonu Cyril C*, Ugwueze Mercy E & Ozioko Chizoba A Department of Pharmaceutics, Universiy of Nigeria,Nsukka, Enugu state, Nigeria.

*Corresponding Author Email: adonucyril@gmail.com

ABSTRACT Phytochemical constituents are responsible for medicinal effects produced by plants. The aim of this study is to evaluate the phytochemical components in ethanol and water extracts of Millettia aboensis, Cuscuta reflexa, Daniella oliveri and Synclisia scabrida. The roots of Millettia aboensis and Synclisia scabrida, aerial parts of Cuscuta reflexa, and leaves of Daniella oliveri were shade dried, pulverized and extracted with alcohol (90% v/v) and sterile distilled water. The phytochemical analyses of these extracts were carried out using standard procedures. The results of phytochemical analysis of both extracts of Millettia aboensis root show that the aqueous extract contained high amount of saponnins, resins, proteins and amino acid and carbohydrate while ethanolic extract has high amount of flavonoids, alkaloids, steroids, and terpenoids. The ethanolic extracts of Cuscuta reflexa, Daniella oliveri and Synclisia scabrida contain copious amount of flavonoids,alkaloids and steroids.

KEY WORDS Phytochemical, Millettia aboensis, Cuscuta reflexa, Daniella oliveria Synclisia scabrida flavonoids,alkaloids.

INTRODUCTION Phytochemicals are non-nutritive chemicals that have protective or disease preventive property. It refers to every naturally occurring chemical presents in plants.

Plants are also the source for many modern pharmaceuticals (drugs). The most important of these phytochemicals are alkaloids, flavonoids, tannins and phenolic compounds [1]. Many of these indigenous plants are used as spices and food plants. Current research has shown that polyphenols contribute to the prevention of cardiovascular diseases, cancers, osteoporosis and antioxidant character with potential health benefits [2-4]. They are known to have beneficial effects on cardio vascular system [5-7]. and have a role in the prevention of neurodegenerative diseases and diabetes mellitu.

Medicinal plants such as Millettia aboensis Cuscuta reflexa, Daniella oliveri and Synclisia scabrida are of great importance to the health of individuals and communities.

Millettia aboensis are small trees of 30–40 feet high and up to 2 feet in girth but usually 12 m high with reddish-brown pubescence on the petioles, branches, International Journal of Pharmacy and Biological Sciences

inflorescence and fruits. They are found commonly in low land rain forest. The flowers are purple in erect woody racemes up to 18 in. long. It has conspicuously rusty-hairy leaves and handsome purple flowers in erect terminal racemes at branch .Almost all the part of Millettia aboensis (uturuekpa) has medicinal properties. The leaf is used by traditional herbalist for general healing including ulcer healing and laxatives while the root is used in treating gastro intestinal disturbances and liver disease. Also the leaf, stem and roots mixed with other plant materials (herbs) is used to cure veneral diseases such as gonorrhoea, syphilis etc.

Cuscuta reflexa is identified by its thin stems appearing leafless, with the leaves reduced to minute scale. It has very low levels of chlorophyll and can photosynthesize slightly. It belongs to the morning glory family, Convolvulaceae, on the basis of the work of the Angiosperm Phylogeny Group.[8] It is found throughout the temperate to tropical regions of the world, with the greatest species diversity in subtropical and tropical regions .It has common names as devil's ringlet, goldthread, hailweed, Adonu Cyril C* et al

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ISSN: 2230-7605 (Online); ISSN: 2321-3272 (Print) Int J Pharm Biol Sci. airweed, hellbine, love vine, devil's guts, devil's hair, angel hair, and witch's hair.

The petroleum ether extract of C. reflexa may be useful in treatment of androgen induced alopecia by inhibiting the enzyme 5alpha-reductase. [9]. The Cuscuta reflexa has been investigated to have the following medicinal effects-bradycardiac [10], antisteroidogenic [11], antihypertensive, psychopharmacological [12] antiviral and anticonvulsant [13] effects. Daniella oliveri is a deciduous tree up to 25-35m tall; bole straight and cylindrical up to 200m in diameter,without buttresses; bark surface smooth. Daniella oliveri is a fast growing tree which occur in tree savanna, bush savanna and in more open grassland on any type of soil, its gum and bark and to a lesser extent the root and the leaves, are widely used in traditional medicine. It has many vernacular names- West African copal tree, Africa copaiba balsam, ilorin balsam, Accra copal, Benin gum copal. Synclisia scabrida Miers(family MENISPERMACEAE) is aslender liane of the rainforest , occurring in Southern Nigeria and Western camerouns to Angola. The root is used in Southeast Nigeria to treat malaria and in some other parts of Nigeria, by local doctors for mental disorders. The study was conducted to analyse ethanol and water extracts of Millettia aboensis, Cuscuta reflexa, Daniella oliveri and Synclisia scabrida.

MATERIALS AND METHODS Plant Material The roots of Millettia aboensis and Synclisia scabrida were harvested from Ehandiagu, while the aerial parts of Cuscuta reflexa, and leaves of Daniella oliveri were got from uzouwani, both in Nsukka, Enugu State, Nigeria. The plants materials were authenticated by Mr. A. Ozioko of Bioresource Development and Conservative Programme (BDCP) Nsukka, Enugu State.

Chemicals: The chemicals used in this study were of analytical grade products of BDH, England nd Sigma Aldrich, Germany.

Extraction of plant materials: The plants parts were shade - dried and pulverized to coarse powder using an electrically operated mill. They were all extracted with 90 % ethanol in soxhlet extractor, concentrated under vacuum. The aqueous extraction was done by 24 h maceration, then filtered and dried.

Phytochemicals analysis The phytochemical analyses of all the plants parts were carried out using standard procedures [14, 15].

Determination of yield of the extracts The percentage yield of the extracts were determined by weighing the coarse plants parts before extraction and after concentration and calculated using the formula

Percentage yield (%) = Weight (g) of the concentrated extracts X 100 Weight (g) of the ground

RESULTS Percentage Yield of the Extracts Table 1: Results of Percentage Yield of all the Extracts Studied.

Plant Part Percentage yield of the extracts (%) Percentage yield of the extract (%) Ethanol Water Millettia aboensis Root 45.93 49.02 Synclisia Scarbrida Root 14.87 19.60 Cascuta reflexa Aerial Parts 6.03 11.33 Daniella oliveri Leaves 17.64 28.94 The percentage yields of all the water extracts are greater than those of the ethanol extracts. The yields of the root extracts of Millettia aboensis are the greatest compared with all other extracts.

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PHYTOCHEMICAL ANALYSIS Preliminary phytochemical investigationThe preliminary phytochemical investigation of the extracts revealed the phytoconstituents presented in Tables 2 & 3 Adonu Cyril C* et al

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ISSN: 2230-7605 (Online); ISSN: 2321-3272 (Print) Int J Pharm Biol Sci. Table 2: Result of Phytochemical Composition of the Roots Extracts of Millettia aboensis and Synclisia scarbrida.

Phytoconstituent Millettia aboensia Synclisis scarbrida EM AM ES AS Flavonoids + +++ ++ +++ Saponins + +++ + +++ Glycosides +++ +++ __ ++ Tannins __ __ ++ __ Carbohydrates ++ +++ __ ++ Steroids ++ + + + Proteins + +++ ++ + Acidic Compounds __ __ __ ___ Alkaloids +++ ++ +++ Resins ___ __ __ + Terpenoids +++ ++ __ + KEY: E.M = Ethanol extracts of Millettia aboensis; AM = Aqueous Extract of Millettia aboensis; ES = Ethanol extracts of Synclisia scarbrida; AS = Aqueous extract of Synclisia scarbrida.

Table 3: Results of Phytochemical Composition of Leaf Extracts of Daniella Oliveri and Aerial Parts of Cuscuta Reflexa.

Phytoconstituent Daniella oliveri Cuscuta reflexa ED AD EC AC Flavonoids Saponins Glycosides Tannins Carbohydrates Steroids Proteins Acidic Compound Alkaloids Resins Terpenoids

+ +++ __ +++ + + __ __ ++ + +

+ +++ __ __ ++ + ++ + ++ ++ +

++ ++ __ __ __ + + + + __ ++

++ ++ __ __ + __ + __ + __ ++

KEY: E.D=Ethanol extracts of Daniella oliveri;A.D=Aqueous extract of Daniella oliveri; EC=Ethanol extracts of Cuscuta reflexa; AC=Aqueous extracts of Cuscuta reflexa

DISCUSSION The results of phytochemical analyses of all the extracts are shown in Tables 2 & 3. The aqueous root extract of Millettia aboensis contained high amount of saponnins, resins, proteins and amino acid and carbohydrate while ethanolic extract has high amount of flavonoids, alkaloids, steroids, and terpenoids. Both extracts have equal amount of glycosides and reducing sugar and absence of International Journal of Pharmacy and Biological Sciences

tannins, acid compounds and fats and oil. The presence of these aforementioned phytocomponents bestow high medicinal importance on Millettia aboensis.

Daniella oliveri leaf water extract revealed the presense of saponins, terpenoids proteins carbohydrates, acidic and lipids components while the ethanol extracts reveals the presence of alkaloids, steroids, and tannins in copious amounts, Adonu Cyril C* et al

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ISSN: 2230-7605 (Online); ISSN: 2321-3272 (Print) Int J Pharm Biol Sci. saponins and flavonoids in moderate amounts and terpenoids, reducing sugars, resins, lipids and carbohydrate in slight amounts. Proteins and glycosides were not found in the ethanol extracts.

The good medicinal qualities possessed by Daniella oliveri leaves are accounted for by the presence of such phytochemicals as alkaloids, terpenoids flavonoids saponins and steroids.

The aqueous and ethanolic extracts of aerial parts of Cuscuta reflexa revealed the presence of steroids, saponins, terpenoids and flavonoids. The presence of these phytochemicals in Cuscuta reflexa accounts [16] [9] for the diuretic , antispasmodic and many pharmacologic activities found with this plant.

The aqueous and ethanolic root extracts of Synclisia scabrida showed the presence of copious amount of alkaloids, flavonoids and tannins. The ethanolic extracts,in addition, showed the presence of proteins, saponins and lipids in moderate to slight amounts.

Many researchers have discovered plants to be rich in secondary metabolites like tannins, alkaloids, flavonoids, phenols, steroids, and volatile oils, which are responsible for their therapeutic activities (17,18).

Most natural alkaloids and their derivatives are used as basic medicinal agents for their analgesic and antibacterial effects.

They exibit marked physiological activity when administered to animals.

Flavonoids are potent water soluble antioxidants and free radical scavengers which prevent oxidative cell damage [19]. Flavonoids in the intestinal tract lower the risk of diseases associated with oxidation (heart disease) [19]. Reports have indicated that flavonoids contributed to the hepatoprotective effect of the plant extracts.

CONCLUSION Both the ethanol and aqueous extracts of Millettia aboensis, Cuscuta reflexa, Daniella oliveri and Synclisia scabrida revealed the presence many phytconstituents with good medicinal qualities. The medicinal value of these plants lies in those components t produce a definite physiological action on the human body.

REFERENCES 1. Lambert J. D, Hong J, Yang G, Liao J and Yang C. S (2005) Inhibition of carcinogenesis by polyphenols:

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evidence from laboratory investigations. The journal Clinical Nutrition. 81, 284–291.

2. Joseph J.A, Shukitt-Hale B and Casadesus G. (2005) reversing the deleterious effects of aging on neuronal communication and behavior: beneficial properties of fruit polyphenolic Compounds. Am. J. Clin.

Nutr. 81, 313– 316.

3. Keen C.L, Holt R.R, Oteiza P.I, Fraga C. G, Schmitz H.H, (2005). Cocoa antioxidants and cardiovascular health. Am. J. Clin. Nutr, 81: 298-303.

4. Sies H, Schewe T, Heiss C, Kelm M, (2005). Cocoa polyphenol and inflammatory mediators. Am. J. Clin.

Nutr, 1: 305-312.

5. Vita J.A (2005) Polyphenols and cardiovascular disease: effects on endothelial and platelet function. Am. J Clin Nutr. 81, 292–297.

6. Scalbert A, Manach C, Morand C, Rémésy C and Jiménez L (2005). Dietary polyphenols and the prevention of diseases.

Critical Rev. The journal of Food Sci. Nutr. 45(4), 287-306.

7. Benjamin L, Shneider S, Philip M. (2008). Pediatric Gastrointestinal Disease, Connecticut publication, PMPH-USA, pp 751. ISBN 1-55009-364-9.

8. Stefanovic', Saša and Olmstead, Richard G. (2004).

Testing the Phylogenetic Position of a Parasitic Plant (Cuscuta, Convolvulaceae, Asteridae): Bayesian Inference and the Parametric Bootstrap on Data Drawn from Three Genomes Syst Biol 53(3): 384-399 doi:10.1080/10635150490445896 9. Pandit S, Chauhan N.S, Dixit V.K., (2008). Effect of Cuscuta reflexa Roxb on androgen-induced alopecia." J Cosmet Dermatol. Sep; 7(3):199-204.

10. Gupta M., Mazumdar U,K, Pal D.K and Bhattacharya S.,( 2003). Anti-Steroidogenic activity of methanolic extract of Cuscuta reflexa roxb. Stem & Corchorus olitorius Linn. Seed in mouse ovary, Ind. J.Exp.Biol.41 (6),6.

11. Singh G.S., and Garg K.N,(1973),Some pharmacological studies on Cuscuta reflexa plant,Ind.J.Pharm(2),344345 12. Pal D, Panda C.,Sinhababu S., Dutta A. and BhattacharyaS., (2003).

Evaluation of psychopharmacologic activity of petroleum ether extract of C. reflexa Roxb. Stem in mice, Pol .Pharm. 60 (6), 481-486.

13. Gupta M., Mazumdar U, K, Pal D.K and Bhattacharya S. and Chakrabarty S ,( 2003). Studies on brain biogenic amines of methanolic extract of Cuscuta reflexa roxb.

Stem & Corchorus olitorius Linn. Seed treated mice,Acta, Pol, Pharm(3),207-210 th 14. Trease G. E. and Evans; (1989). Pharmacognosy, 13 Edition. ELBBS Oxford university press, London, UK. Pp 245-26

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15. Harbone J.B,(1998). Phytochemical methods (3 Edition) Thompson Science, 2-6 Boundary Row, London,UK. PP 107-150.

16. Sharma S, Hullati k.k., Prasanna S. M.,Kuppast I.J.,Sharma P,(2009). Comparative Study of Cuscuta reflexa Cassytha filiformis for diuretic acivity.Phcog Res 1: 327-30 17. Cowman M.W. (1999). Plant products as antimicrobial agents, Clin. Microb, Rev, 12 564 – 582

18. Rabe T. and Vanstoden J. (2000). Isolates of an antimicrobial sequiterpeniod from Warburgie salitaris.

J. Ethnopharmacol, 2: 113-118.

19. Salah N, Miller N. J, Pagange G, Tburg L, Bolwell G. P, Rice E, Evans C, 1995).

Polyphenolic flavonoids as scanvengers of aqueous phase radical as chain breaking antioxidant. Arch. Biochem. Bioph.; 2: 339 – 346.

*Corresponding Author: Adonu Cyril C* Email: adonucyril@gmail.com

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International Journal of Pharmacy and Biological Sciences- ISSN: 2321-3272 (Print) IJPBS | Volume 3 | Issue 2 | APR – JUN | 2013 | 06-11 Research Article – Biological Sciences

Central Nervous System Anomalies in Teaching Hospital Hyderabad Rameswarapu Suman Babu1 & Sujatha Pasula2 1

Associate professor , Department of Anatomy, Mediciti Institute of Medical Sciences, Ghanpur, Ranga Reddy District-501401.A.P, India 2*, Assistant Professor Department of Biochemistry, D D Institute of Medical Science and Research, Thiruvalluri, Tamilnadu *Corresponding Author Email: drsujathapasula@gmail.com

ABSTRACT Objective: The present study was undertaken to explore the incidence of central nervous system anomalies in present geographical area. Method: Study includes 2000 consecutive pregnancies that have come for check up in second and third trimester with clinically relevant CNS malformations detectable by ultrasonography. Results: Analysis has revealed that there are 74 fetal anomalies in 34 fetuses with CNS anomalies and 18 fetuses with neural tube defects. Conclusion: The overall incidence of congenital foetal anomalies in the present study is 1.7 % which needs to supplementation of folic acid and vitamin B12 during peri-conceptional period and early diagnosis by available protocol.

KEY WORDS Central nervous system anomalies, Neural tube defect, Anencephaly

INTRODUCTION Congenital anomalies of the fetuses are a great concern since time immemorial. Etiological factors for these anomalies are plenty. Environmental pollution, radiation, exposure to different chemicals, teratogenic drugs are some of the reasons for these anomalies.

There is a strong correlation between the congenital anomalies of the fetuses and chromosomal abnormalities, either structural or numerical. Central nervous system (CNS) malformations are some of the most common of all congenital abnormalities (1) The Central nervous system is of ectodermal in origin and appears as the neural plate at the middle of the 3rd week. After the edges of the plate become folded, the neural folds approach each other in the midline to fuse into the neural tube. The cranial end closes approximately at day 25, and the caudal end closes at day 27. The CNS then forms a tubular structure with a broad cephalic portion, the brain, and a long caudal portion, the spinal cord. The various developmental anomalies of the central nervous system are anencephaly, menigomylocele, spina bifida, encephalocele and hydrocephalus.

An opening in the spinal cord or brain is called Neural tube defect which amounts to about 1–2 cases per 1000 births, the most frequent CNS malformations.

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Anencephaly is one of most common anomaly characterized by failure of the cephalic part of the neural tube to close. As a result, the vault of the skull does not form, leaving the malformed brain exposed.

Later, this tissue degenerates, leaving a mass of necrotic tissue(2).The incidence of intracranial abnormalities with an intact neural tube is uncertain as probably most of these escape detection at birth and only become manifest in later life. Long-term follow-up studies suggest however that the incidence may be as high as one in 100 births (3). Gaining hands on experiences show that, birth defects pose multifarious social, economic and cultural problems as well as mental trauma to the whole humanity. It is widely noticed that, many mothers are not aware of the impact of factors in causing congenital defects in the fetuses. For better understanding of the etiological factors of congenital anomalies, knowledge of embryology, teratology, clinical genetics and diagnostic ultrasonography is very important (4,5). Highly advanced imaging techniques such as 3-D and 4-D ultrasound have been helping largely in diagnosing and treating birth defects in the fetus during the antenatal period. It is strongly advised that, antenatal ultrasonography has to be conducted compulsorily for a minimum of two times to all antenatal mothers.

Detection of the CNS anomalies in the prenatal period Rameswarapu Suman Babu & Sujatha Pasula

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ISSN: 2230-7605 (Online); ISSN: 2321-3272 (Print) Int J Pharm Biol Sci. is very important for the prognosis of fetus because they result in abortions and still births.

Further, early detection of CNS anomalies identification enables us to do corrective surgeries for these fetuses. As a result we can give a healthy and normal baby to the mother. Identification of these abnormalities also has become easy with the invention of ultrsonography without causing any discomfort either to the mother or to the fetus. Also medical management which includes termination of pregnancy and counseling the eligible couple will result in betterment of the society by attaining eugenics. All these reasons created interest to study the CNS anomalies and to show the incidence of their abnormalities. Anencephaly, menigomyelocele, spina bifida, encephalocele are categorized under neural tube defects.

MATERIALS AND METHODS The present study was undertaken to explore the incidence of CNS fetal anomalies during the antenatal period. The present study is based on the ultrasonographic diagnosis of 2000 pregnancies in 2011-12 which were studied in the department of Radiology, government maternity hospital attached to Osmania medical college, Hyderabad from. Cases were selected from women in second and third trimesters with age of 20-35 years who came for obstetric services.

The indications for scanning were hydramnios, history of suspected anomaly on clinical examination, determination of EDD, multiple pregnancy, oligohydramnios, history of antenatal bleeding in second and third trimesters, aged mothers, young mothers, history of anomalies in other siblings, for fetal well being and routine. Women were selected irrespective of age, parity and from all socio-economic

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classes. A detailed history of anomalies in other siblings, history of consanguinity, age of the mother and socio-economic status were also noted.

RESULTS On 2000 consecutive antenatal ultrasound examinations it is reported that detection of 34 CNS anomalies in screening of these subjects. All cases of congenital anomalies were arranged anomalie wise and tabulated. Where ever possible sonographic print outs along with clinical photographs which were followed with termination were taken, these babies were injected formalin for fixation and later autopsy was done in Osmania medical college. Detailed history, period of gestation and results of present study were noted in proforma.

Total number of detected fetal anomalies by ultrasound out of 74 congenital anomalies, 34 were with CNS anomalies.Babies with NTD were 18 in incidence i.e 9 /1000 births. Following classification is made based on anomalies observed.

A) Anencephaly was seen in 10 cases (13.51%) in USG and planned for termination .On examination of fetuses 2 cases with meningocele (Figure-1), 2 cases associated with absence of supra renal glands (Figure2), 1 case with diaphrmatic hernia and others with only anencephaly were observed. B) Spina bifida with meningocele was seen in 1 case with lubosacral meningocele (Figure-4), 1 case with myelocele. C) Encephalocele was observed in 2 cases (Figure-3), 1 case with associated deformity of heart and lungs. D) Hydrocephalus seen in 8 cases (Figure-5). E) Multisystem involvement in 10 cases ,1 case with hydrocephalus, meningocele, omphalocele ,renal agenesis and other with spinal bifida with dextrocardia, exompholos(Figure-5).

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Maternal age in years 20 - 25 26 - 30 30-35 >35 Total(34)

Anencephaly 4 2 2 2 10

Table-1: In relation to Maternal age Spina bifida with Other CNS anomalies Meningo\ myelocele 1 2 3 1 3 2 2 2 9 7

Associated multiple anomalies 2 4 1 1 8

More cases of CNS anomalies especially NTD is more in younger age group i.e 20-25 yrs.

2 trimester rd 3 trimester

8 2

Table-2: In relation to Gestational age Spina bifida with Other CNS Meningo\ myelocele anomalies 4 2 4 4

Total (34)

10

8

Gestational age nd

Anencephaly

Most of the cases were detected in 2

Degree of consanguinity 0

Associated multiple anomalies 7 3

6 nd

10

trimester when they come for antenatal check up.

Table-3: In relation to degree of consanguinity.

Anencephaly Other anomalies 8 10

1

2

2

2

1

5

3

2

4

Total (34)

13

21

There is significance of consangunity in the group.

Gender

Anencephaly

Female Male Total (34)

7 3 10

Table-4: In relation to Gender of fetus Spina bifida with Meningo\ Other CNS myelocele anomalies 5 4 5 2 10 6

Associated multiple anomalies 2 6 8

Females are more affected than males

CNS anomalies Anencephaly Meningo\ myelocele Spina bifida Encephalocele Hydrocephalus Multi system involvement Total (74)

Table-5: The distribution of the CNS anomalies No. of fetuses affected 10 4 2 2 8 8 34

Percentage 13.51 5.41 2.70 2.70 10.81 10.81 45.94 %

In the present study 2000 USG scans recorded out of which 34 CNS anomalous cases were detected. Thus the all over incidence o f the CNS anomalies was 1.7 %.

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Figure-1: Showing anencephaly with meningocele.

Figure-2: Showing encephalocele and associated deformity of heart and lungs.

Figure-3: Lumbo sacral meningocele

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Figure-4: Hydrocephalus

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Figure-5: Multi system involvement with spina bifida with dextrocardia, exompholos

DISCUSSION There are several reports that suggested the incidence, particularly pattern of congenital central nervous system (CNS) anomalies which may vary in different geographical locations (6–9). However, the extent to which such variations reported were attributable to differences in genetic inheritance, environmental factors or diagnostic accuracy is uncertain. Studies on the incidence and pattern of different types of congenital abnormalities can provide valuable information for planning health care services, including preventive programs for better future of mankind.

In this study as per the data (Table-1) collected, the age of the mother does not carry much weightage because most of the mothers come under eligible couple age and none below18 years. But genetic factor shows significance here, though karyotyping and gene studies were not conducted (Table-2).

From Table-3 there is history of consanguinity in 10 out of 34 cases with anomalies and in the rest of the cases environmental factors probably play a role in causing CNS defects which correlates with Al-Gazali et al (10).

Prevalence is high in females compared to males in our study which is in correlation with Mahadevan et.al (Table-4) (11).

The CNS anomalies are detailed in Table No.5 which show 34 fetuses with central nervous system anomalies out of which neural tube defects (NTD) were diagnosed in 18/2000 and among them anencephaly was seen in 10 /2000. Reddi rani et al(12) reported 21 CNS cases of which 14 were with NTD in polyhydramnios selected by USG and also International Journal of Pharmacy and Biological Sciences

Sania et al (13) showed 46/3310 cases of NTD .

Dhapate S.S et al (14) and Bala kumar et al (15) studied CNS anomalies using USG showed incidence of 48% and 40% respectively which is in much correlation to our study which is 45.94% .Our study evenly correlates with other studies also (16,17).

Increased frequency of NTD could be due to maternal malnutrition and low socioeconomic status could be responsible for it. Advances in genetics and molecular biology have led to a better understanding the control of central nervous system (CNS) development. It is possible to classify CNS abnormalities according to the developmental stages at which they occur. The careful assessment of patients with these abnormalities is important in order to provide an accurate prognosis.

Counseling should be given to couples with consanguinity and nutritional deficiencies, regarding recurrent risk of genetic defects and educating the mothers about environmental factors responsible for occurrence of neural tube defects help to decrease its occurrence. In addition, these families provide a good opportunity to map and identify the responsible genes in order to improve our knowledge about the mechanisms of normal and abnormal development of the CNS. Hence it is said that “Prevention is better than cure”.

Health services planning should be to supplementing folic acid and vitamin B12 during peri-conceptional period and early diagnosis by available protocol.

CONCLUSION The prevalence of CNS anomalies in the present population at risk is 1.7%. A tool used here is Rameswarapu Suman Babu & Sujatha Pasula

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ISSN: 2230-7605 (Online); ISSN: 2321-3272 (Print) Int J Pharm Biol Sci. antenatal ultrasound method which has an advantage of detecting neural tube defects more accurately. In conclusion, this study has provided a detailed analysis of CNS abnormalities in a population with consanguinity. Careful and detailed analysis of these anomalies is required for accurate diagnosis and genetic advice.

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ACKNOWLEDGEMENTS This study was undertaken in the department of Anatomy, Osmania Medical College, (OMC) Hyderabad, under the guidance of Dr. T.K. Rajasree, Professor of Anatomy and with supervision by Dr. M. Ravinder, Professor and Head, Department of Anatomy, OMC. Our sincere thanks also to Dr. S. Sreelatha, Associate Professor and to Dr. Krishnaveni, Professor of Radiology, OMC. The authors would also like to thank HOD and Professor B.R.Potturi (Mediciti Institute of Medical Sciences) for his valuable advice in giving the final shape to this manuscript.

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Pietrzyk JJ, Grochowski J, Kanska B. CNS malformations in the Krakow region: birth prevalence and seasonal incidence during 1979–1981. Am J Med Genet 1983: 14: 181–188.

Al-Gazali LI, Sztriha L, Dawodu A, Bakir M, Varghese M, Varady E, Scorer J, Abdulrazzaq Ym, Bener A, Padmanabhan R. Pattern of central nervous system anomalies in a population with a high rate of consanguineous marriages. Clin Genet 1999: 55: 95– 102. mahdevan et.al :neural defects in pondicherry :Indian J pediatr 2005;72(7);557-559.

Reddi rani et al.clinical and USG evaluation of polyhydramnious :Journal of Obs and Gyn of India :2003:53(2);145-148 Sania tanveer etal .Incidence of risk factors for NTD in Peshawar ;Journal of Medical sciences :2008:6(1)1-4 Dhapate S.S ,etal:Early diagnosis of anenecephaly value of USG in rural area :Journal of anatomical society of India 2007:56(2)4-7.

Balakumar K.Major anotommical fetal anomalies in northern kerela ; Journal of Obs and Gyn of India 2007:57(4) 311-315.

Sharada B.menasinkai et al:A study of neural tube defects :J anat soc india: 2010: 59(2)162-167 Nakling J et al Adverse obstetric outcome in fetuses that are smaller than expected at second trimester routine ultrasound examination.:Acta Obstet Gynecol scand. 2005 Nov;84(11):1042-8

*Corresponding Author: Rameswarapu Suman Babu & Sujatha Pasula Email:drsujathapasula@gmail.com

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International Journal of Pharmacy and Biological Sciences- ISSN: 2321-3272 (Print) IJPBS | Volume 3 | Issue 2 | APR – JUN | 2013 | 12-14 Review Article – Pharmaceutical Sciences

INTRODUCTION TO DIFFERENT TYPES OF IMMUNO ANALYZERS AND ITS USEFULNESS IN DIFFERENT TYPES OF IMMUNO ASSAYS H.Patel*1, B.Patel1 & S.Parmar2 1

P.G.Deparment of Pharmaceutical Sciences, Sardar Patel University,V.V.Nagar,Gujarat,India 2 P.G.Deparment of Pharmaceutical Sciences,Sardar Patel University,V.V.Nagar,Gujrat,India

*Corresponding Author Email: hiral.pharma@yahoo.in

ABSTRACT ImmunoAnalyzerare used for the estimation of metabolic activity of vitamins, hormones, for performing immuno assays, detection of blood clot etc. by using Human plasma. Immuno Analyzer like, Abbott AxSym is used for efficient therapeutic drug monitoring and for serology tests. It has the capability to add tests while the instrument is operating. The throughput rates ranges from 80 tests/hour up to 120-tests / hour average. In addition to automation, the IMx system offers rapid assay times and calibration curve storage without sacrificing clinical efficacy. The fully automated IMx immunoassay analyzer was used to develop a system for the detection of IgG and IgM antibodies to rubella virus for immune status screening and diagnosis of primary infections. Antihuman IgG or IgM antibody coupled to alkaline phosphatase enzyme followed by methylumbelliferyl phosphate substrate was used to detect the presence or absence of antibodies specific to the antigens on the solid phase. To evaluate the efficacy of the IMx rubella IgG assay, immune status screening was performed with a clinical patient population of 501 sera. When compared to an IgG specific enzyme immunoassay and passive hemagglutination assay the agreement was greater than 99%. The IMx rubella IgM assay was utilized to determine the presence of rubella specific IgM antibodies in 462 sera. These results were compared to IgM specific enzyme immunoassay results and also demonstrated greater than 99% agreement. Serocon version following rubella vaccination of susceptible individuals was demonstrated by IgG and IgM antibody responses as early as two weeks post vaccination. By the use of LIASON Immunoanalyzer Measles and Mumps IgG Assays can be rapidly performed.

The BioPlex 2200 Antiphospholipid Syndrome (APLS) IgG, IgM, and IgA kits are the first and only fully-automated, randomaccess multiplex flow immunoassays for the semi-quantitative detection of IgG, IgM, and IgA antibodies to cardiolipin (CL) and beta-2 glycoprotein I (β2GPI). Vitamins assays can also be performed very rapidly. The FDA-cleared STA-StaclotdRVV Screen and Confirm are fully automated assays for detecting lupus anticoagulants (LA) in patient plasma by the diluted Russell viper venom method. The Ecarin Chromogenic Assay (ECA) is an enhancement of the Ecarin Clotting Time (ECT), the reference method for determining direct thrombin inhibitors (DTIs) such as argatroban. DPC Immulite is a Diagnostic Products Corporation (now a division of Siemens) manufactured immunoassay system, offering fully integrated operation.

KEY WORDS Abbott AxSym, IgG, IgM, IgA, Liason, BioPlex 2200, STA-Staclotd RVV

INTRODUCTION Different types of immuno analyzers: 1) AbottAxSym: AbottAxSymImmuno Analyzer AbottAxSym which is manufactured by Abbott Diagnostics has a high capacity and reliable chemistry analyzer designed to meet the needs of today’s laboratories.

Abbott AxSym is used for efficient therapeutic drug monitoring and for serology tests. This thirdInternational Journal of Pharmacy and Biological Sciences

generation immunoassay system with broad testing features has random, continuous-access, and STAT processing capabilities. Abbott AxSYM utilizes three technologies such as fluorescence polarization immunoassay, microparticle enzyme immunoassay, and the new ion-capture immunoassay. With the use of AxSym device, you can run a wide range of immunodiagnostic tests at a time. Moreover, it has the capability to add tests while the instrument is

H.Patel* et al

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ISSN: 2230-7605 (Online); ISSN: 2321-3272 (Print) Int J Pharm Biol Sci. operating. The throughput rates ranges from 80 tests/hour up to 120-tests / hour average.

 

AbottAxSym 2) Immulite 2500 Immunoassay System:

         

    Immulite 2500 Immunoassay System Immulite 2500 Immunoassay System is manufactured

by Siemens Medical Solutions Diagnostics. It is approved by US FDA, CE. Its throughput rate is 200 Tests/Hour and process time is 15, 35 and 65 minutes.

Available tests are Anemia, Allergy, Cardiovascular, Congenital Assays, Fertility, Infectious Disease, Metabolic Function, Oncology, Therapeutic Drug Monitoring and Thyroid. It is continuous, random, and STAT access. It can Clot and liquid level detection.

FEATURES  Unique user interface software for enhanced operation  Onboard refrigeration  Automatic monitoring of reagents, supplies and waste  Minimal attendance, easy access and limited maintenance  Provides a loading capacity of up to 200 patient samples  Reduces sample handling by allowing use of clinical chemistry instrument racks  Allows continuous and immediate access to patient samples

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

Real-time instrument monitoring via the Internet, 7 days a week, 24 hours a day Online QC management from any Internetequipped computer Throughput of up to 200 tests/hour 24 resident assays Proprietary onboard dilution Automatic re-assay of out-of-range samples Reflex testing for additional clinical information Primary, secondary and micro tube sampling LIS interface, including host query Remote-test ordering via LIS STAT capability Open architecture to interface with lab automation or work cell configurations via the SMS Clot detection Proprietary wash technique for high sensitivity Third-generation assays Extensive menu encompassing routine and esoteric assays, for faster clinical diagnostic information Reagents, controls, and consumables may be available.

Different types of assays: 1) LIAISONMeasles and Mumps IgG Assays: The LIAISON Measles and Mumps IgG Assays are based upon the chemiluminescent immune assay. This assay used to detect IgG antibodies to measles and mumps viruses in human serum. This helps in determination of serological status to measles and mumps virus. It gives first result in 35 minutes and throughput of 90 results/hour, these chemiluminescent immune assays complete the MMRV IgG assay panel.

2) 25- hydroxy Vitamin D Assay: Diazyme's 25-Hydroxy Vitamin D assay measures the true total 25-hydroxy vitamin D and the sum of both D3 + D2. It has a wide dynamic range with improved precision and eliminates time-consuming washing steps. This assay reduces testing time and cost. The test is user-friendly and can be run manually. Easily adapted for use on a wide range of fully automated microtiter plate readers, making it suitable for use in laboratories of all sizes and testing needs.

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ISSN: 2230-7605 (Online); ISSN: 2321-3272 (Print) Int J Pharm Biol Sci.

3) STA-StaclotdRVV Screen and Confirm assay for detecting lupus anticoagulants: The FDA-cleared STA-StaclotdRVV Screen and Confirm are fully automated assays. It detects lupus anticoagulants (LA) in patient plasma by the diluted Russell viper venom method. Each reagent is barcoded for the ease of use. The STA Staclotd RVV Screen uses a reagent with a low phospholipid concentration; it will enhance its sensitivity and prolong the clotting time.

The STA StaclotdRVV Confirm uses a reagent with a high phospholipid concentration. This neutralizes the LA present in the plasma, which will shorten the clotting time. These tests are not affected by contact factor deficiencies, factor VIII, and IX deficiencies, or other specific inhibitors, as well as by samples containing therapeutic levels of heparin. STA Staclotd RVV Screen and Confirm in combination with Staclot LA are complimentary in fulfilling the recommendations established by ISTH. 4) Ecarin Chromogenic Assay: The Ecarin Chromogenic Assay (ECA) enhances the Ecarin Clotting Time (ECT).It is the reference method for determining direct thrombin inhibitors (DTIs) such as argatroban. For this many high-risk patients are required to minimize dangerous side effects.

5) BioPlex 2200 APLS IgG, IgM, and IgA Kits for diagnosis of antiphospholipid syndrome:

The BioPlex 2200 Antiphospholipid Syndrome (APLS) IgG, IgM, and IgA kits are the first fully-automated, random access multiplex flow immunoassays for the semi-quantitative detection of IgG, IgM, and IgA antibodies to cardiolipin (CL) and beta-2 glycoprotein I (β2GPI). These kits helpsin the diagnosis of antiphospholipid syndrome (APS). These kits consolidate six traditional single-analyte tests into one disease-state panel. The through put of this assay is up to 200 tests / hour. These kits also offer results in 45 minutes. The BioPlex 2200 APLS IgG, IgM, and IgA help in autoimmune and infectious disease tests on the BioPlex 2200 system.

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www.abbott.com international.abbottdiagnostics.com en.wikipedia.org/wiki/abbott-axsym Smith J., OsikowiczG., AbbottAxsym random & continuous access Immuno assay system for Improved work flow in the clinical laboratory, in the journal of clinical chemistry, Oct 1993, 39(10), 2063-2069. www.diasorin.com/measles...mumps www.captodayonline.com/immuno H Tuokko,Comparision of non specific reactivity in indirect and reverse Immuno assays for measles & mumps Immunoglobulin M antibodies, Journal of clinical microbiology,1984. www.biopacific.net/Diazyme-25-OH www.aacc.org

*Corresponding Author: H.Patel * Email: hiral.pharma@yahoo.in

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