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Indian Journal of Experimental Biology

 

ISSN : 0019-5189

CODEN : IJEB (A6) 42(10)  945-1036 (2004)

VOLUME 42

NUMBER 10

OCTOBER 2004

 

CONTENTS 

Review Article

 

Liposomes as adjuvant for anti-mycobacterial vaccine development

949

Indu Verma, Rajesh Pandey & G K Khuller

 

 

 

Papers

 

Influence of malaria parasite (P. vivax) on erythrocyte aggregation: A study based on dynamic imaging and analysis

955

Sanjay Jayavanth & Megha Singh

 

 

 

Effect of fly ash inhalation on biochemical and histomorphological changes in rat lungs

964

V Suresh Kumar, U Mani, A K Prasad, Kewal Lal, V Gowri & Abhishek Gupta 

 

[IPC Code: Int. Cl.7 A61]

 

 

 

Biokinetics of 65Zn in liver and whole body and its bio-distribution in nickel treated protein deficient rats

969

Pardeep Sidhu, M L Garg & D K Dhawan

 

[IPC Code: Int. Cl.7 A61K]

 

 

 

Secretion of metastasis related gangliosides by mouse B16-melanoma in circulation in vivo and in culture media in vitro

976

S Saha & K C Mohanty 

 

 

 

Antitumour activity of some plants from Meghalaya and Mizoram against murine ascites Daltonís lymphoma

981

G Rosangkima & S B Prasad

 

[IPC Code: Int. Cl.7  A61P]

 

 

 

Antioxidant effect of Boerhavia diffusa L. in tissues of alloxan induced diabetic rats

989

M Amarnath Satheesh & L Pari

 

[IPC Code: Int. Cl.7 A61P]

 

 

 

Effect of propolis extract on acute carbon tetrachloride induced hepatotoxicity

993

Sangeeta Shukla, Monika Bhadauria & Anjana Jadon

 

[IPC Code: Int. Cl.7 A61P]

 

 

 

Modulatory potential of Spirulina fusiformis on testicular phosphatases  in Swiss albino mice against mercury intoxication

998

 Preeti Suman Saxena & Madhu Kumar

 

 

Toxicity of Nerium indicum Miller seed extract on bandicoot rat,
Bandicota bengalensis Gray

1003

K Saravanan, S Senthilkumar, M Elayaraja & B Suresh

 

 

 

Histology of vitamin A induced ectopic limbs and normal hind limbs in tadpoles of Polypedates maculatus

1007

Pravati K Mahapatra, Sagarika Mishra & Sushil K Dutta

 

 

 

Notes

 

Evaluation of secondary poisoning of difethialone, a new second-generation anticoagulant rodenticide to Barn owl, Tyto alba Hartert under captivity

1013

K Saravanan & R Kanakasabai

 

 

 

Enzymatic changes induced by some organophosphorous pesticides in female rats

1017

Sukhdeep Kaur & C K Dhanju 

 

 

 

Production of solasodine by Solanum laciniatum using plant tissue culture technique

1020

Pooja Bhatnagar, Madhurima Bhatnagar, Amarjit K Nath & D R Sharma

 

[IPC Code: Int. Cl.7 A01H1/00]

 

 

 

Protective effect of vilva juice on glycoconjugate levels in experimentally induced constipation in rats

1024

R Padmini, K E Sabitha & C S Shyamala Devi

 

[IPC Code: Int. Cl.7 A61P]

 

 

 

Mini transposon vector mediated foreign gene expression in Mesorhizobium huakuii subsp. rengei

1028

D Balachandar, S Kannaiyan, H Ono & Y Murooka

 

 

 

Identification of glandular (preputial and clitoral) proteins in house rat (Rattus rattus) involved in pheromonal communication

1032

G Archunan, S Kamalakkannan, S Achiraman & R Rajkumar 

 

 

 

Announcements

1036

ICMR Awards and Prizes for 2002 and 2003; Ramanbhai Foundation 2nd International Symposium; National Symposium on Biodiversity and Insect Pest Management

 

Author Index
 
Keyword Index

 

 

 

Indian Journal of Experimental Biology

Vol. 42, October 2004, pp. 949-954

 

 

Review Article

  

Liposomes as adjuvant for anti-mycobacterial vaccine development

Indu Verma, Rajesh Pandey & G K Khuller

 

Mycobacteria are intracellular pathogens that invade and reside inside the macrophages. Recent advances in controlled delivery systems for vaccines such as liposomes have sparked a renewed interest in their potential application for the prevention of mycobacterial infections. The versatility of liposomes in the incorporation of hydrophilic/hydrophobic components, their non-toxic nature, biodegradability, biocompatibility, adjuvanticity, induction of cellular immunity, property of sustained release and prompt uptake by macrophages, makes them attractive candidates for the delivery of antigens. This review focuses on liposome research in the area of mycobacterial diseases and highlights how the various mycobacterial components may be exploited as powerful antigens with liposomes as adjuvants.

 

 

 

Indian Journal of Experimental Biology.

Vol 42, October 2004, pp. 955-963

 

Papers

  

Influence of malaria parasite (P. vivax) on erythrocyte aggregation:
A study based on dynamic imaging and analysis

Sanjay Jayavanth & Megha Singh

 

Malaria due to P. vivax (PV) is prevalent in many countries. The present work is aimed to determine the cell-cell interaction through formation of aggregates under dynamic conditions. Blood samples are obtained from patients (n=11) suffering from PV malaria, and the normal subjects (n=10) in test tubes containing citrate phosphate dextrose (10:1.4), as an anticoagulant. The signature analysis of infected erythrocytes shows significant alterations in their shape and membrane. For aggregation analysis, erythrocyte suspension in plasma at hematocrit 5%, was placed in a glass chamber and mounted vertically on the stage of the video-microscope system. The aggregate images thus acquired show erythrocytes adhering with each other to form mash-like structures. With increase in parasitaemia, the erythrocytes show hyper-aggregation compared to that of normal cells. By processing of the sequence of recorded images during sedimentation, the various aggregation parameters are obtained. These parameters show that the formed aggregates are compact which produce distinct changes in sedimentation pattern with significantly higher sedimentation velocity compared to that in healthy blood samples. These changes in malaria could partly be responsible for alteration in blood flow through microcirculatory system.

 

 

 

Indian Journal of Experimental Biology

 Vol. 42, October 2004, pp. 964-968

 

Effect of fly ash inhalation on biochemical and histomorphological changes in rat lungs

V Suresh Kumar, U Mani, A K Prasad, Kewal Lal, V Gowri & Abhishek Gupta

 

Effect of respirable fly ash particles inhalation on lungs of rats was investigated by exposing them to respirable aerosols of size classified power plant fly ash at average concentrations of up to 14.4Ī1.77 mg/m3 for 4 hr/day for 28 consecutive days. A remarkable increase was found in blood eosinophil counts of fly ash exposed animals. Biochemical indicators of pulmonary damage viz. lactate dehydrogenase (cytoplasmic enzyme used as a measure of cell injury), γ-glutamyl transferase (Clara cell damage) and alkaline phosphatase (potential measure of Type II cell secretions) in broncho alveolar lavage fluid (BALF) of fly ash exposed group showed significant elevation. Clumping of fly ash particles in the lungs was observed as evidenced by fly ash ladened macrophage accumulation in the alveolar region. The results suggest a damage, local inflammation and remodelling of lung as indicated by hypertrophy and hyperplasia. These changes reflect the toxic effects of the fly ash inhalation.

 

 

 

Indian Journal of Experimental Biology

 Vol. 42, October 2004, pp. 969-975

 

Biokinetics of 65Zn in liver and whole body and its bio-distribution
in nickel treated protein deficient rats

Pardeep Sidhu, M L Garg & D K Dhawan

 

This study was designed to determine the effect of nickel treatment on biological half-lives of 65Zn in whole body and liver as well as on distribution of 65Zn in different organs of protein deficient rats. Nickel sulfate at a dose level of 800mg/l in drinking water was administrated to normal control as well as to protein deficient rats for 8 weeks. A significant increase was found in fast and slow components of biological half lives of 65Zn in whole body and only fast component in liver of protein deficient rats. Interestingly, slow component in whole body and fast component in liver of nickel treated protein deficient rats were not different from normal controls though they were significantly elevated in protein deficient rats. On the other hand, slow component of 65Zn was also not altered in nickel treated protein deficient rats, which however, was significantly decreased in nickel treated rats. Protein deficiency led to a marked elevation in per cent uptake of 65Zn in brain and caused significant depression in liver, kidney and intestine. However, uptake of 65Zn in brain showed a significant depression in nickel treated rats, whereas the uptake was elevated in brain in nickel treated protein deficient rats. In conclusion, protein deficient conditions seem to be playing a dominant role in context with the distribution of 65Zn in different organs when nickel is administered to protein deficient rats. However nickel alone is seen to cause adverse effect on the distribution of 65Zn.

 

 

 

Indian Journal of Experimental Biology

 Vol. 42, October 2004, pp. 976-980

  

Secretion of metastasis related gangliosides by mouse B16-melanoma
in circulation in vivo and in culture media in vitro

S Saha & K C Mohanty

 

Mouse B16LuF1 melanoma cells of lower metastatic potential to lung were treated in vitro with same concentration (50ĶM) of gangliosides prepared from plasma of mice bearing lung metastasis of B16LuF5, B16LuF9 or B16LuF10 melanoma cell lines of increasing metastatic potential to lung(LuF1< LuF5 < LuF9< LuF10) and injected to normal mice through tail vein. The number of metastatic tumor nodules formed in lung increased gradually in mice receiving B16LuF5, B16LuF9 and B16LuF10-ganglioside-treated B16LuF1 cells compared to mice receiving B16LuF1 cells without any ganglioside treatment. Similarly, mouse B16LuF1 melanoma cells treated in vitro with 50 ĶM concentration of gangliosides prepared from spent culture media of B16LuF5, B16LuF9 or B16LuF10 cells cultured in vitro were injected to normal mice through tail vein. The number of metastatic tumor nodules formed in lung increased gradually in mice receiving B16LuF5, B16LuF9 and B16LuF10-ganglioside-treated B16LuF1 cells compared to mice receiving B16LuF1 cells without any ganglioside treatment. The results indicated that metastasis-associated gangliosides present in plasma and culture media of B16-melanoma of increasing metastatic potential to lung enhanced metastatic potential of B16LuF1 cells. The increasing concentration of metastasis-associated gangliosides present in plasma and in culture media of B16-melanoma of increasing metastatic potential possibly determined increase in metastatic potential of B16LuF1-melanoma cells.

 

 

 

Indian Journal of Experimental Biology

Vol. 42, October 2004, pp. 981-988

 

Antitumour activity of some plants from Meghalaya and Mizoram
against murine ascites Daltonís lymphoma

G Rosangkima & S B Prasad

 

Total five plants, three from Mizoram (Dillenia pentagyna, Ageratum conyzoides, Blumea lanceolaria) and two from Meghalaya (Potentilla fulgens, Taxus baccata) were studied for their antitumour activity against murine ascites Daltonís lymphoma (DL) in vivo. Only three plants showed the different magnitude of antitumour activity. Out of these three plants, the antitumour activity was maximally observed with the methanol extract of the stem bark of D. pentagyna as compared to the aqueous extract of the roots of A. conyzoides and aqueous extract of the root of P. fulgens. An increase in glutathione levels in Daltonís lymphoma cells was observed during tumour growth. Changes in glutathione and protein levels were also investigated in the liver and Daltonís lymphoma cells of tumour-bearing mice following the treatment with the extract of D. pentagyna which showed the highest antitumour activity as compared to the other two plant extracts. Glutathione in the liver and DL cells of treated tumour-bearing mice was found to be decreased. The protein concentration in liver and DL cells decreased mainly at 96 hr of treatment. It may be concluded that the natural product of D. pentagyna promises to be more active against Daltonís lymphoma than others and the decrease in glutathione level may be one of the important steps in resulting this antitumour effect.

 

 

 

Indian Journal of Experimental Biology

 Vol. 42, October 2004, pp. 989-992

 

Antioxidant effect of Boerhavia diffusa L. in tissues of alloxan induced diabetic rats

M Amarnath Satheesh & L Pari

 

Administration of B. diffusa leaf extract (BLEt; 200 mg/kg) for 4 weeks resulted in a significant reduction in thiobarbutric acid reactive substances and hydroperoxides, with a significant increase in reduced glutathione, superoxide dismutase, catalase, glutathione peroxidase and glutathione ĖS- transferase in liver and kidney of alloxan induced diabetic rats. The results suggest that BLEt has remarkable antidiabetic activity and can improve antioxidant status in alloxan induced diabetic rats.

 

 

 

Indian Journal of Experimental Biology

 Vol. 42, October 2004, pp. 993-997

 

Effect of propolis extract on acute carbon tetrachloride induced hepatotoxicity

Sangeeta Shukla, Monika Bhadauria & Anjana Jadon

 

Ethanolic extract of propolis was administered to rats intoxicated by carbon tetrachloride. Administration of bolus dose of CCl4 (1.5 ml/kg, ip) resulted in elevation of serum transaminases and serum alkaline phosphatase activities. Levels of hepatic lipid peroxidation were significantly increased. On the contrary, there was significant decrease in hepatic reduced glutathione level. The propolis extract (100 and 200 mg/kg, po) exhibited recoupment in both pre- and post-treatment (prophylactic and curative studies) of biochemical changes induced by CCl4. The post treatment of 200 mg/kg, po extract showed most significant hepatoprotective effect. Histopathological studies showed damage in hepatocytes and disturbed chord arrangement after toxicant administration. Propolis extract (200 mg/kg, po) was found to be more effective in restoring CCl4 induced histopathological alterations.

 

 

 

Indian Journal of Experimental Biology

 Vol. 42, October 2004, pp. 998-1002

 

Modulatory potential of Spirulina fusiformis on testicular phosphatases
in Swiss albino mice against mercury intoxication

Preeti Suman Saxena  & Madhu Kumar

 

Administration of mercuric chloride (HgCl2; 5.0 mg/kg body weight) to male Swiss albino mice resulted in significantly higher levels of testicular acid phosphatase (ACP) and alkaline phosphatase (ALP) activities as compared to control. In combination group where S. fusiformis (800 mg/kg body weight) was given before and after HgCl2 treatment, the mercury induced toxicity reduced in terms of decreased levels of ACP and ALP activities in the testis. The animal treated with only Spirulina did not show any alteration in ACP and ALP values. It is suggested that oral administration of Spirulina can modulate mercury induced testicular toxicity.

 

 

 

Indian Journal of Experimental Biology

Vol. 42, October 2004, pp. 1003-1006

  

Toxicity of Nerium indicum Miller seed extract on bandicoot rat,
Bandicota bengalensis Gray

K Saravanan, S Senthilkumar, M Elayaraja & B Suresh

 

Crude extract (12.5 ml/kg) of N. indicum seed gave 100% mortality of B. bengalensis. Humanness assessment study revealed that this plant orgin chemical caused low pain and sufferings to the target pests.

 

 

 

Indian Journal of Experimental Biology

 Vol. 42, October 2004, pp. 1007-1012

 

Histology of vitamin A induced ectopic limbs and normal hind limbs
in tadpoles of Polypedates

Pravati K Mahapatra, Sagarika Mishra & Sushil K Dutta

 

Marked histological similarities were observed between normal and vitamin A induced ectopic limb buds of P. maculatus. However, close association of nephric tubule and lateral plate mesoderm, as seen in normal hind limb bud does not seem to be essential for ectopic limb development. The ectopic limbs tend to develop in pairs.

 

 

 

Indian Journal of Experimental Biology

Vol. 42, October 2004, pp. 1013-1016

 

maculatus

 

 Evaluation of secondary poisoning of difethialone, a new second-generation anticoagulant rodenticide to Barn owl, Tyto alba Hartert under captivity

K Saravanan & R Kanakasabai

 

Secondary toxcity of difethialone to Barn owl (Tyto alba) has been investigated. Difethialone was fed to owls for successive periods of 1 (phase 1), 3 (phase 2) and 6 (phase 3) days via-rodenticide dosed rats. The owls survived after the treament of rodenticide on phase 1 and phase 2 experiments but they died during phase 3 experiment. The results suggest that the difethialone could cause more secondary toxicity to owls.

 

 

 

Indian Journal of Experimental Biology

Vol. 42, October 2004, pp. 1017-1019

 

Enzymatic changes induced by some organophosphorus pesticides in female rats

Sukhdeep Kaur & C K Dhanju

 

An evaluation of toxic effects of three organophosphorus pesticides viz. monocrotophos, methyl parathion and dimethoate given orally daily for 90 days was done in terms of enzymatic changes in plasma and liver of female albino rats. A significant decrease was observed in the level of esterases in plasma with all the three pesticides. The activity of acid and alkaline phosphatases and aminotransferases increased significantly in plasma and significantly or marginally in liver with these pesticides. The results are thus indicative of the cellular toxicity of these organophosphates even after their subchronic administration in low doses for a long period.

 

 

Indian Journal of Experimental Biology

Vol. 42, October 2004, pp. 1020-1023

 

 Production of solasodine by Solanum laciniatum using plant tissue culture technique

Pooja Bhatnagar, Madhurima Bhatnagar, Amarjit K Nath & D R Sharma

  

Leaf and hypocotyl explants of 15 days old aseptically grown seedlings of Solanum laciniatum were cultured on MS medium supplemented with NAA (2 mg/l) and kinetin (0.5 mg/l) for callus initiation. For maintenance and proliferation of callus MS medium supplemented with 2,4-D (1 mg/l) and kinetin (0.5 mg/l) was used. The growth of the calli derived from hypocotyls increased with time of incubation and remained almost constant after 45 days. The solasodine content in callus culture was maximum after 30 days of incubation. Addition of L-arginine in the medium (50-150 mg/l) increased growth as well as chlorophyll content in the callus culture. The solasodine content also increased up to 1.2 to 1.4 times in these cultures. High frequency shoot regeneration was obtained in MS medium having BA (4 mg/l) and IBA (0.25 mg/l). For shoot multiplication, MS medium having BA (4mg/l) was used. Shoots rooted on the same medium. Organogenesis promoted solasodine accumulation in the cultures. Regenerated shoots yielded higher solasodine content than undifferentiated as well as organogenic callus. Solasodine contents in the regenerated shoots was found to be 10 times higher than the callus culture and approached towards the field grown plants. Thin layer chromatography revealed the presence of three compounds. The most predominant spot (Rf 0.789) corresponded to the reference solasodine.

 

 

 

Indian Journal of Experimental Biology

Vol. 42, October 2004, pp. 1024-1027

 

Protective effect of vilva juice on glycoconjugate levels in experimentally
induced constipation in rats

R Padmini, K E Sabitha & C S Shyamala Devi

 

Efficacy of vilva, a polyherbal formulation was evaluated in morphine induced constipated rats. Vilva juice, at a dose of 1.5 ml/100 g body wt was given orally for a period of 7 days. Morphine sulfate was injected to induce constipation on 8th day, 45 min before the experiments. Protein bound glycoconjungates were estimated in intestinal tissue. Altered levels of glycoconjugates were maintained at near normalcy when pretreated with vilva juice in morphine induced rats. Histological changes were observed in the colon tissue. The damage to crypts of Liberkunn in constipated rats were found to be reduced in vilva pretreated rats. Vilva, thus, offered significant protection against morphine induced constipation by way of augmenting mucus secretion.

 

 

 

Indian Journal of Experimental Biology

Vol. 42, October 2004, pp. 1028-1031

 

Mini transposon vector mediated foreign gene expression in
Mesorhizobium huakuii subsp. rengei

D Balachandar, S Kannaiyan, H Ono & Y Murooka

 

Among the transposable elements, mini-Tn5 transposon vector has proven to be of greater utility for insertion mutagenesis of variety of Gram negative bacteria. The mini-Tn5 vector containing promoter less egfp gene and gentamycin resistant gene was used for the present study. The transposon vector was introduced to M. huakuii from E. coli S17 by conjugation. The conjugants were screened for stable expression of egfp both in free-living and in nodules of Astragalus sinicus. The result showed that the conjugant #3 showed stable expression of green fluorescent both in free-living and bacteroid stage. The visualization of sym plasmid of wild strain and conjugants showed that conjugant #3 had a fragmentation of large sized plasmid into two but without affecting the nodulating ability. These results clearly indicated that mini-Tn5 vectors (Transposon vectors) the best alternate tools for plasmid vectors for integration of foreign genes in chromosomal DNA or symbiotic plasmid and expression, both in free-living and bacteroid stage of Rhizobium.

 

 

 

Indian Journal of Experimental Biology

 Vol. 42, October 2004, pp. 1032-1035

 

 Identification of glandular (preputial and clitoral) proteins in house rat (Rattus rattus)
involved in pheromonal communication

G Archunan, S Kamalakkannan, S Achiraman & R Rajkumar

 

Proteins (18-20 kDa) belonging to lipocalin family have been reported to act as carriers for ligands binding to pheromones in mouse urine, pig saliva, hamster vaginal fluid and human sweat, that are involved in pheromonal communication. As the preputial gland is a major pheromonal source, the present study was aimed to detect the specific protein bands (around 18Ė20 kDa) in the preputial and clitoral glands of the house rat, R. rattus. The amount of protein was higher in preputial gland of the male than that of female (clitoral) gland. A 20 kDa protein was noted in male and female glands; however, the intensity of the band was much higher in male than in female. In addition, 70, 60, 35 kDa bands, identified in male preputial gland, were absent in females. The presence of higher concentration of glandular proteins in the male preputial gland suggests that male rats may depend more on these glandular proteins for the maintenance of reproductive and dominance behaviours. The results further suggest that these glandular proteins (20 kDa) may act as a carrier for ligand binding.

 

 

Author Index

 

Achiraman S

1032

Jadon Anjana

993

Ono H

1028

Archunan G

1032

Jayavanth Sanjay

955

 

 

 

 

 

 

Rajkumar R

1032

Balachandar D

1028

Kamalakkannan S

1032

Rosangkima G

981

Bhadauria Monika

993

Kanakasabai R

1013

 

 

Bhatnagar Madhurima

1020

Kannaiyan S

1028

Sabitha K E

1024

Bhatnagar Pooja

1020

Kaur Sukhdeep

1017

Saha S

976

 

 

Khuller G K

949

Saravanan K

1003, 1013

Devi C S Shyamala

1024

Kumar Madhu

998

Satheesh M Amarnath

989

Dhanju C K

1017

Kumar V Suresh

964

Saxena Preeti Suman

998

Dhawan D K

969

 

 

Senthilkumar S

1003

Dutta Sushil K

1007

Lal Kewal

964

Sharma D R

1020

 

 

 

 

Shukla Sangeeta

993

Elayaraja M

1003

Mahapatra Pravati K

1007

Sidhu Pardeep

969

 

 

Mani U

964

Singh Megha

955

Garg M L

969

Mishra Sagarika

1007

Suresh B

1003

Gowri V

964

Mohanty K C

976

 

 

Gupta Abhishek

964

Murooka Y

1028

Verma Indu

949

 

 

 

 

 

 

 

 

Nath Amarjit K

1020

 

 

 

 

Keyword Index

 

Abnormal tail

1007

 Gangliosides

976

Parasitaemia

955

Adjuvant

949

Glandular proteins

1032

Pheromones

1032

Alloxan diabetes

989

Glutathione

981

Plasmodium vivax

955

Anticoagulant

1013

Glycoconjugates

1024

Polypedates maculatus

1007

Antigen

949

 

 

Propolis

993

Antitumour activity

981

Hepatoprotection

993

Protein deficiency

969

 

 

 

 

 

 

B16-melanoma

976

Inhalation

964

Rat

964, 1017,
1032

Bandicota bengalensis

1003

 

 

Rhizobium

1028

Barn owl

1013

L-Arginine

1020

Rodenticide

1013

Biopesticide p10

1003

Lipid peroxidation

989

 

 

Boerhavia diffusa

989

Lipocalin

1032

Secondary poisoning

1013

 

 

Liposome

949

Seed extract

1003

Callus culture

1020

Lung

964

Shape parameters

955

Carbon tetrachloride (CCl4)

993

 

 

Solanum laciniatum

1020

Conjugation

1028

Macrophage

949

Solasodine

1020

Constipation

1024

Malaria

955

Spirulina

998

 

 

Mercury intoxication

998

sym Plasmid

1028

Daltonís lymphoma

981

Metastatic potential

976

 

 

Difethialone

1013

Modulatory potential

998

Testicular phosphatases

998

Dillenia pentagyna

981

Morphine sulfate

1024

Transposon vectors

1028

 

 

 

 

Tuberculosis

949

Ectopic limb

1007

Nerium indicum

1003

Tumour

976

Enzymic antioxidants

989

Nickel

969

Tyto alba

1013

Erythrocyte aggregation

955

Normal hind limb

1007

 

 

 

 

 

 

Video-microscopic images

955

Fly ash

964

Organogenesis

1020

Vilva juice

1024

 

 

Organophosphates

1017

Vitamin A

1007

 

 

 

 

 

 

 

 

 

 

65Zn

969