The CRISPR-Cas9 induced CCR5 Δ32 mutation as a potent gene therapy methodology for resistance to HIV-1 variant: a review.

Human Immunodeficiency Virus (HIV) has continuously been the greatest epidemic for humanity over a period spanning almost five decades. With no specific cure or treatment available to date despite extensive research, the C-C Chemokine Receptor 5, Delta 32 (CCR5 Δ32) allele genetic point mutation plays an imperative role in the prevention of acquired immunodeficiency syndrome (AIDS). This comprehensive study aims to review the induction of the homozygous recessive deletion genotype using the Clustered Regularly Interspaced Short Palindromic Repeats, Cas 9 Enzyme (CRISPR-Cas9), and hematopoietic stem cell transplantation under positive selection pressure for active immunity in seropositive patients' populations as the phenotype. A methodology is proposed to trigger a significant increase in the expression of Delta 32 beneficial mutant alleles within controlled modern healthcare facilities utilizing totipotent stem cells through somatic gene therapy. It acts upon two dysfunctional CCR5 genes, translating mutant G protein-coupled co-receptors, whose primary function is similar to that of C-X-C Motif Chemokine receptor 4 (CXCR4), by blocking the entry of viral RNA into the CD4+ T helper lymphocytes, halting infection and seizing viral life cycle. This modification is endemic in Northern Europe, where it naturally pertains to the Caucasian descent population samples in the form of polymorphism, p (X=0.01), where X is the probability of frequency of complete immunity against HIV-1 in population samples. The epigenetics of the single nucleotide polymorphism (SNP) are analyzed as they play a significant role in immunity distribution. Furthermore, a comparative analysis within the ethical boundaries of CRISPR-Cas9 is conducted to discuss the practical aspects and challenges of the presented methodologies and treatment alternatives. Additionally, the study assembles all available data and summarizes preexisting research while providing a promising solution to this ethical dilemma. Finally, a methodology is devised to answer the question of whether the variant-specific epidemic of AIDS caused by HIV-1 can be cured via artificially inducing immunity by CRISPR-Cas9.

Unusual High Hardness and Load-Dependent Mechanical Characteristics of Hydrogenated Carbon-Nitrogen Hybrid Films.

Mechanical components are exposed to a rigorous environment in a number of applications including engineering, aerospace, and automobiles. Thus, their service lifetime and reliability are always on the verge of risk. Protective coatings with high hardness are required to enhance their service lifetime and minimize the replacement cost and waste burden. Hydrogenated amorphous carbon including nitrogen-incorporated films, that are commonly deposited by plasma-enhanced chemical vapor deposition, are widely used for commercial protective coating applications. However, their mechanical hardness still falls into the moderate hard regime. This needs to be substantially enhanced for advanced applications. Here, we report the synthesis of very hard nanostructured hydrogenated carbon-nitrogen hybrid (n-C:H:N) films. The optimized n-C:H:N film displays a hardness of about 36 GPa, elastic modulus of 360 GPa, and reasonably good elastic recovery (ER) of 62.7%. The mechanical properties of n-C:H:N films are further tailored when nitrogen pressure is tuned during the growth. The realized remarkably improved mechanical properties are correlated with the films' structural properties and experimental growth conditions. We also conducted density functional theory calculations that show the trend for the elastic modulus of the amorphous carbon films with varying nitrogen concentrations matches well with experimentally measured values. Finally, we probed load-dependent mechanical properties of n-C:H:N films and found an anomalous behavior; some of the mechanical parameters, for instance, ER, reveal an irregular trend with indentation load, which we explain in the framework of the film-substrate composite concept. Overall, this work uncovers many unknown and exciting mechanical phenomena that could pave the way for new technological developments.

Electrosurgery Dissection versus Sharp Dissection: Effect on Early Postoperative Wound Complications in Modified Radical Mastectomy.

Modified radical mastectomy (MRM) is still the most commonly performed operation for breast cancer, despite the trends toward breast-conserving treatment. Since 1970s, electrosurgery (also known as electrocautery, diathermy) has been a widespread surgical tool to raise flaps and excise the breast specimen in order to perform a bloodless mastectomy. Use of diathermy has been well blamed for wound complications. To prevent undue delay in the adjuvant treatment, it is important to minimize the surgical complications. This quasi experimental study was conducted in the Department of Surgery, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka, Bangladesh to evaluate the effects of electrosurgery and sharp dissection on early postoperative wound complications in MRM. 46 patients were included in the study (23 in each group). Data analysis done in SPSS version 23.0 and 'p' value <0.05 considered significant at 95% confidence interval. In Electrosurgery Dissection (ED) group mean duration of drainage was 7.8±1.2 days and in Sharp Dissection (SD) group 6.4±1.0 days (p value 0.000). Mean total drainage in ED group found 1082±287ml and in SD group 693±194ml (p value 0.000). Seroma formation found 7(30.4%) in ED group and 3(13.0%) in SD group (p value 0.004). Wound dehiscence found 5(21.7%) in ED group and 2(8.7%) in SD group (p value 0.013). Flap necrosis rate was 4(17.4%) and 1(4.4%) in ED and SD group respectively (p=0.003). Demographic and clinical variables were similar or differences were not statistically significant in two groups. No difference found in operating time and wound infection rate in two groups. The result of the study showed that, electrocautery dissection caused early postoperative wound complications more than the sharp dissection.

Pigtail Catheter in the Management of Liver Abscess.

Amoebic liver abscess is a common condition in Bangladesh. Though conservative treatment plays a major role, refractory patients can be successfully treated with minimally invasive approach. This prospective study was carried out to evaluate the prospect of sono-guided percutaneous catheter drainage for liver abscess patients not responding to conservative treatment. This study was done at Dhaka Medical College Hospital from January 2005 to June 2006 among patients with liver abscess that were treated conservatively outside and got admitted in different surgery and medicine units. Included 35 patients were evaluated clinically and through relevant investigations. Sono-guided percutaneous pigtail catheter was introduced for drainage and patients were followed up for two weeks on average at hospital indoor by amount of drainage, clinical improvement and serial follow up USG. After discharge from the hospital, all patients were followed up monthly and assessed clinically and ultrasonographically up to two months on outdoor basis. Thirty five patients underwent pigtail catheter drainage that was refractory to conservative treatment or needle aspiration. Following insertion of catheter patients who were pyrexic, fever subsided in two to three days and never back during the postoperative period and follow up. Antimicrobials were changed according to the report of the culture and sensitivity of the aspirate that was done routinely. Maximum total (4300 ml) amount of pus was drained in a patient. Average duration of catheter drainage in this study was 8 days. With a very low morbidity and zero mortality rates, minimum treatment costs and early return to regular life style proved this minimally invasive procedure to be a rational treatment option for amoebic liver abscess in selective cases.

Time-dependent tegumental surface changes in juvenile Fasciola gigantica in response to triclabendazole treatment in goat.

Triclabendazole (TCBZ), the anthelmintic drug active against both mature and immature liver flukes, was used to investigate the effect of in vivo treatment on the tegumental surface of juvenile Fasciola gigantica. Five goats were infected with 150 F. gigantica metacercariae each by oral gavage. Four of them were treated with single dose of TCBZ at 10mg/kg at four weeks post-infection. They were euthanized at 0 (untreated), 24, 48, 72 and 96h post treatment. Juvenile flukes were manually retrieved from the goat livers and processed for scanning electron microscopy. In control flukes, the anterior region was adorned with sharply pointed spines projecting away from the surface, while in the posterior region, spines become shorter and narrower, loosing serration and with the appearance of distinct furrows and papillae. The dorsal surface retained the same pattern of surface architecture similar to that of ventral surface. Flukes obtained from 24h post-treatment did not show any apparent change and were still very active. However, there were limited movements and some blebbing, swelling, deposition of tegumental secretions and some flattening displayed by the flukes of 48h post-treatment. All the worms were found dead 72h post-treatment and showed advanced level of tegumental disruptions, consisting of severe distortion of spines, sloughing off the tegument to expose the basal lamina, formation of pores and isolated patches of lesions. By 96h post-treatment, the disruption was extremely severe and the tegument was completely sheared off causing deeper lesions that exposed the underlying musculature. The disruption was more severe at posterior than anterior region and on ventral than dorsal surface. The present study further establishes the time-course of TCBZ action in vivo with 100% efficacy against the juvenile tropical liver fluke.

Sub-100 nm patterning of TiO2 film for the regulation of endothelial and smooth muscle cell functions.

Previous studies have shown that fundamental cell functions such as adhesion, proliferation, and morphology are regulated by the interaction of cells with basement membrane nano- and micron- scale surface topography. By taking the basement membrane as a guiding principle, the surface can be designed with biophysical cues in the form of surface topographies to modulate the cellular functions of vascular endothelial and smooth muscle cells, which are crucial for vascular diseases. Titanium-based materials whose surfaces are covered by a thin layer of titanium dioxide (TiO2) are utilized in several regenerative medicine applications such as vascular prosthetics. The utilization of TiO2-covered materials makes it essential to understand the interaction of cells with the TiO2 layer to control the cell response. While it has been shown by means of patterned micro- and nano-topography that it is important to regulate cell functions on non-metallic materials, it would be of interest in the field of regenerative medicine to study the cell response on patterned TiO2 surface layers. Previous studies have mostly focused on studying the cell response on random micro- and nano-roughened metallic and metal oxide surfaces as it is challenging to fabricate patterned TiO2 surface layers. Here, the biocompatibility of a method that is capable of the rational patterning of a continuous TiO2 surface layer of sub-100 nm resolution scale using thermally curable resin-based nanoimprinting was studied. The responses of human umbilical vein endothelial cell (HUVEC), such as proliferation, morphology and focal adhesions, and smooth muscle cell (SMC) proliferation and morphology to the nano-patterned TiO2 layer were investigated. Overall, the TiO2 layer with surface nano-gratings showed enhanced proliferation of HUVECs, while it significantly lowered the proliferation of SMCs as compared to the unpatterned control. The HUVECs and SMCs were shown by topography to be sensitive to the 70 nm gratings as evident by the regulation of proliferation and cell shape. A significantly lower focal adhesion density was found of HUVECs on TiO2 nano-gratings, while a significantly higher average focal adhesion size of HUVECs was seen on TiO2 nano-wells and nano-gratings, compared to the unpatterned controls.

A rabbit eye model for in vivo transformation of progenetic metacercariae of Clinostomum complanatum into ovigerous adult worms.

Clinostomum complanatum is a digenetic trematode that causes yellow grub disease in some fish species and also shows zoonotic potential by sporadically infecting humans. In this study, progenetic metacercariae of C. complanatum were obtained from the fish Trichogaster fasciatus, and were aseptically placed in conjunctival incisions made in the superior and inferior fornices of the eye of rabbits, which served as the experimental hosts. Worms were harvested without necropsy of the host on days 4 and 8 post infection, to observe in vivo transformation of the progenetic metacercariae into ovigerous adult worms. The worms appeared to cause minimal damage to the host although they were tenaciously attached. In vivo maturation was evident by the development of the vitellaria, enlargement of gonads, the presence of a large number of shelled eggs in a distended uterus and ramifications of the intestinal caeca. Obtaining mature ovigerous worms without sacrificing the host clearly gives the rabbit eye model an advantage over those described previously. Due to the relative advantage of the short time required for maturation and the prolific egg production by C. complanatum, it is suggested that this host-parasite system could be used as an excellent model for classroom teaching of trematode biology and to investigate the cues involved in in vivo transformation and host-parasite interactions.

Diagnostic evaluation of supraclavicular lymphadenopathy.

Patients presented with the supraclavicular lymphadenopathy in the medicine department have a strong suspicion of serious illness like tuberculosis, sarcoidosis, toxoplasmosis and malignancy of lymphnode, blood, lung, upper GIT, breast, ovary, testes, and other sites of body. This prospective type of observational study carried out in the indoor and out patient department of medicine of Mymensingh Medical College Hospital over a period of 6 month from April 2011 to September 2011 to diagnose the causes of supraclavicular lymphadenopathy. Patient of either sex, 18 years or above presented with supraclavicular lymphadenopathy were included. Biopsy or FNAC were done. The study showed that mean age of the patient of supraclavicular lymphadenopathy that finally diagnosed as malignant was 49.7 years and that of non malignant was 33.7 years. Male patient have suffered more (60%) from malignant disease than that of female patient (40%). Discrete, hard, non tender either fixed or non fixed supraclavicular lymphadenopathy was found malignant (18 of 18 cases, 100%) and discrete, firm, tender lymphnode were found non malignant (5 of 5 cases, 100%). Increased frequency (11 of 28, 39.3%) of granulomatous inflammation from the tuberculoid lymphadenitis were found among the patient undergone supraclavicular lymphnode biopsy. FNAC result was also of simillar type and finally it was found that frequency of tuberculosis (20 of 53, 37.7%) was highest and bronchial carcima was the second most frequent diagnosis (14 of 53, 26.4%). This study showed that supraclavicular lymphadenopathy is associated mostly with serious disease like tuberculosis and malignancy.

Proteomics and in silico approaches to extend understanding of the glutathione transferase superfamily of the tropical liver fluke Fasciola gigantica.

Fasciolosis is an important foodborne, zoonotic disease of livestock and humans, with global annual health and economic losses estimated at several billion US$. Fasciola hepatica is the major species in temperate regions, while F. gigantica dominates in the tropics. In the absence of commercially available vaccines to control fasciolosis, increasing reports of resistance to current chemotherapeutic strategies and the spread of fasciolosis into new areas, new functional genomics approaches are being used to identify potential new drug targets and vaccine candidates. The glutathione transferase (GST) superfamily is both a candidate drug and vaccine target. This study reports the identification of a putatively novel Sigma class GST, present in a water-soluble cytosol extract from the tropical liver fluke F. gigantica. The GST was cloned and expressed as an enzymically active recombinant protein. This GST shares a greater identity with the human schistosomiasis GST vaccine currently at Phase II clinical trials than previously discovered F. gigantica GSTs, stimulating interest in its immuno-protective properties. In addition, in silico analysis of the GST superfamily of both F. gigantica and F. hepatica has revealed an additional Mu class GST, Omega class GSTs, and for the first time, a Zeta class member.

Direct nanoimprint lithography of Al2O3 using a chelated monomer-based precursor.

Nanostructuring of Al2O3 is predominantly achieved by the anodization of aluminum film and is limited to obtaining porous anodized aluminum oxide (AAO). One of the main restrictions in developing approaches for direct fabrication of various types of Al2O3 patterns, such as lines, pillars, holes, etc, is the lack of a processable aluminum-containing resist. In this paper, we demonstrate a stable precursor prepared by reacting aluminum tri-sec-butoxide with 2-(methacryloyloxy)ethyl acetoacetate, a chelating monomer, which can be used for large area direct nanoimprint lithography of Al2O3. Chelation in the precursor makes it stable against hydrolysis whilst the presence of a reactive methacrylate group renders it polymerizable. The precursor was mixed with a cross-linker and their in situ thermal free-radical co-polymerization during nanoimprinting rigidly shaped the patterns, trapped the metal atoms, reduced the surface energy and strengthened the structures, thereby giving a ~100% yield after demolding. The imprinted structures were heat-treated, leading to the loss of organics and their subsequent shrinkage. Amorphous Al2O3 patterns with line-widths as small as 17 nm were obtained. Our process utilizes the advantages of sol-gel and methacrylate routes for imprinting and at the same time alleviates the disadvantages associated with both these methods. With these benefits, the chelating monomer route may be the harbinger of the universal scheme for direct nanoimprinting of metal oxides.

Observations on in vitro and in vivo antimicrofilarial effects of Bishop's weed (Trachispermum ammi).

The antimicrofilarial efficacy of Trachispermum ammi extacts in vitro and in vivo using Setaria cervi as a model, was investigated. T. ammi seed extracts were prepared using different solvents (with increasing order of polarity of the solvent) including petroleum ether, diethyl ether, chloroform, ethyl acetate, acetone, ethanol, and methanol. The extracts were tested for in vitro antimicrofilarial activity. The ethanolic and the methanolic extracts showed maximum activity in causing flaccidity in the microfilariae. The extracts were potent even at concentrations as low as 5 µl/ml. When orally administered to experimentally infected rats, the extracts eliminated circulating microfilariae within 2 weeks. It is inferred that the antimicrofilarial molecule(s), are polar in nature. They induce flaccidity in the microfilariae, by possibly inhibiting monoamine oxidase. This communication supplements the ethnopharmacological information for the use of T. ammi as an antihelminthic, and indicates that T. ammi could be used as a potential source of antimicrofilarial drugs.

Direct imprinting of high resolution TiO(2) nanostructures.

We demonstrate a different approach to direct nanoimprint lithography of oxides, in particular TiO(2), using the metal methacrylate route which not only gives very high resolution ( approximately 20 nm) but also provides yields of approximately 100% over areas > 1 cm x 1 cm. TiO(2) was imprinted using a polymerizable liquid 'TiO(2) resin' consisting of a mixture of titanium methacrylate, ethylene glycol dimethacrylate, and azobis-(isobutyronitrile). The resin underwent free radical polymerization when imprinted using a silicon mold at 110 degrees C with pressures as low as 10 bar. Polymerization strengthens the imprinted structures, thereby giving approximately 100% yield after demolding. Heat-treatment of the imprinted structures at 400 degrees C resulted in the loss of organics and their subsequent shrinkage ( approximately 75%) without the loss of integrity or aspect ratio, and converted them to TiO(2) nanostructures as small as approximately 20 nm wide. Furthermore, our method demonstrates that large imprinted areas of sub-100-nm features can be achieved by sub-micron molds which translate into huge cost savings with the added flexibility of direct patterning of urinary as well as multi-component oxides.

Upper abdominal mass with diagnostic dilemma.

A 32 years old gentleman presented with a lump occupying the epigastric, left hypochondriac and umbilical region for the last 3 months which gradually increased in size & dull aching pain. He also complained of low grade fever and early satiety. On examination, he had a lump on upper abdomen which was irregular, firm, mildly tender, lobulated surface, restricted mobility, dull on percussion. USG examination of HBS revealed hepatomegaly and SOL in liver. CT abdomen revealed soft tissue mass in left upper abdomen. FNA revealed cells of mesenchymal origin. Per operatively a large lobulated, myxomatous highly vascular mass arising from the fundal part of the stomach was found and removed. Histopathology report comments about two differential diagnosis-leiomyosarcoma and gastrointestinal stromal tumour. CD117 immunostaining confirmed it was gastrointestinal stromal tumor (GIST). Post operatively patient was treated by Imatinib and is now under regular follow up.

Trapped charge dynamics in a sol-gel based TiO(2) high- k gate dielectric silicon metal-oxide-semiconductor field effect transistor.

We have studied the response of a sol-gel based TiO(2), high k dielectric field effect transistor structure to microwave radiation. Under fixed bias conditions the transistor shows frequency dependent current fluctuations when exposed to continuous wave microwave radiation. Some of these fluctuations take the form of high Q resonances. The time dependent characteristics of these responses were studied by modulating the microwaves with a pulse signal. The measurements show that there is a shift in the centre frequency of these high Q resonances when the pulse time is varied. The measured lifetime of these resonances is high enough to be useful for non-classical information processing.

Characterization of the glutamate dehydrogenase activity of Gigantocotyle explanatum and Gastrothylax crumenifer (Trematoda: Digenea).

Glutamate dehydrogenase (GLDH) (EC 1.4.1.3) is a ubiquitous enzyme, which is present at the protein and carbohydrate metabolism crossroads. The enzyme activity was investigated in biliary and rumen amphistomes, Gigantocotyle explanatum and Gastrothylax crumenifer, respectively, infecting the Indian water buffalo Bubalus bubalis. The enzyme activity was consistently higher in G. explanatum as compared to G. crumenifer, where NAD(H) was utilized as coenzyme and the pH optima was recorded at 8. The K(m) and V(max) values for α-ketoglutarate were 2.1 mM and 9.09 units in G. explanatum, whereas 3.03 mM and 1.90 units in G. crumenifer, respectively. Among the allosteric modulator nucleotides, AMP, ADP, ATP, GMP, CMP and UMP, only AMP enhanced GLDH activity in G. crumenifer while ADP was stimulatory in G. explanatum. The amino acid leucine stimulated the GLDH activity in both the amphistomes while alanine was stimulatory only in G. crumenifer. Pronounced interspecific differences in response to different metabolic inhibitors like diethyldithiocarbamate, semicarbazide hydrochloride and mercurial ions were also observed. The osmotic stress alters the enzyme activity, particularly in hypertonic saline the GLDH activity increased significantly (p < 0.01) in G. explanatum, while insignificant effects were observed in rumen dwelling G. crumenifer. Histoenzymology revealed region/tissue specific distribution of GLDH with prominent staining in tissues like vitellaria, lymph system and tegument/subtegument, thus showing specific distribution of GLDH indicating differential metabolic state. Such intergeneric differences in GLDH activity could also be a consequence of occupying different microenvironments within the same host.

TiO2 patterning using electro-hydrodynamic lithography.

An increasing number of technologies require the fabrication of micro- and nanostructures over large areas. Soft lithographic methods are gaining in popularity for the manufacture of low-cost micrometre and sub-micrometre structures. Increasingly, these methods developed to structure organic resists can also be used to pattern inorganic materials. Here we introduce a simple lithographic technique that is able to pattern ceramic TiO micro- and nanostructures with high fidelity. Our method makes use of an electrohydrodynamic (EHD) film instability that is controlled by a laterally modulated electric field. A spin-coated film of a stabilized metal alkoxide precursor material was patterned using EHD lithography followed by a heat treatment at 400 °C to yield crystalline TiO micropatterns. Our technique is rather general and can be extended to a number of single- and multicomponent oxide systems.

Partial purification and characterization of Gigantocotyle explanatum somatic antigens.

Soluble extracts of Gigantocotyle explanatum, isolated from the liver of buffalo Bubalus bubalis were fractionated on Sephadex G-200 columns. Nine major fractions referred to as F1, F2, F3, F4, F5, F6, F7, F8 and F9 were separated. Each fraction was tested by ELISA for antigenicity using sera from G. explanatum-infected field buffaloes. Fractions F1 and F2 were highly antigenic, F3, F4, F6 and F7 were moderately antigenic and F5, F8 and F9 were poorly antigenic. Analyses by SDS-PAGE revealed that each fraction comprised several polypeptide(s) in the molecular weight range of <29 to >205 kDa. Results of Western blotting indicated that not all polypeptides which appeared in the SDS-PAGE were antigenic. The antigenic molecules of each fraction were mostly in the low molecular weight range of <14 to >94 kDa with the polypeptides in the range of >14, 14, 18, 21-25 and 34-36 kDa.

Single crystals of single-walled carbon nanotubes formed by self-assembly.

We report the self-assembly of single crystals of single-walled carbon nanotubes (SWCNTs) using thermolysis of nano-patterned precursors. The synthesis of these perfectly ordered, single crystals of SWCNTs results in extended structures with dimension on the micrometer scale. Each crystal is composed of an ordered array of tubes with identical diameters and chirality, although these properties vary between crystals. The results show that SWCNTs can be produced as a perfect bulk material on the micrometer scale and point toward the synthesis of bulk macroscopic crystalline material.

Analysis of potential antigens of protoscoleces isolated from pulmonary and hepatic hydatid cysts of Bubalus bubalis.

The aqueous soluble proteins of the protoscoleces of Echinococcus granulosus isolated from the pulmonary and hepatic hydatid cysts of Bubalus bubalis were partially purified on Sephadex G-200 column. The isolated fractions were tested for their antigenicity by immunodot using the sera collected from experimentally infected puppies during the prepatent period of infection. Six protein profiles (F1-F6) were recovered from both the isolates but the polypeptide recovery of each fraction of the two isolates were some what different, particularly protein concentration of F5 in the liver isolate was greater than the lung isolate. Contrary to this, the concentration of F1 polypeptides of the lung origin protoscoleces is greater as compared to that of the liver. In addition, some antigenic dissimilarity has also been observed in the two isolates. A weak IgG response against F1, F2 and F6 polypeptides was observed in the 4th day post infection (p.i.) sera. With the age of infection the response against these antigens increased as revealed by the intensity of the spot in immunodot analysis. Our studies show that the differences in the elution profile and antigenic profile of the lung and liver isolates, as revealed by gel filtration and immunodot analysis, might be either due to the influence of the microhabitat or these may be two different strains. Further studies are certainly required in this direction.

Partial purification and characterization of Gastrothylax crumenifer somatic antigens.

The soluble extracts of Gastrothylax crumenifer isolated from the rumen of buffalo (Bubalus bubalis) were fractionated on a Sephadex G-200 column. A total of eight major fractions (F1, F2, F3, F4, F5, F6, F7, and F8) were separated from the whole homogenate of G. crumenifer, and each of these fractions was tested for their antigenicity by ELISA against rabbit hyperimmune sera. It was observed that F1, F2, F3 and F4 were highly antigenic, F6 and F7 were moderately antigenic and F5 and F8 were poorly antigenic. The individual fractions analysed after SDS-PAGE and Western blotting indicated that the antigenic fractions of G. crumenifer are of low molecular weight, in the range of <14-50kDa, and predominant antigenic components which were evident in most of the Sephadex profiles were of Mr 15, 18, 19, 23-24 and 28-32kDa.