Molecular Biology and Biotechnology

Phenylhydrazine (PHZ), a potent chemical causes toxicity on various tissues at various levels. Administration of phenylhydrazine mainly causes haematotoxicity which leads to the haemolytic anemia. In mammals PHZ induced anemia increased the iron absorption in spleen, liver and duodenum and finally iron metabolism was altered. Local demand and supply of Fe would increase erythropoeitic activity of the spleen so the size of spleen was increased that create the splenomegaly. PHZ induced anemia activate immune response which triggers phagocytosis in the spleen and liver. Apart from this administration of PHZ interfere the binding of erythropoietin (EPO) with erythropoietin receptors (EPOR) so that JAK-STAT would be affected.PHZ also showed genotoxic effect by creating single strand DNA damage.

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A fingerprint of maize hybrid DHM 121 and its parental inbreds was generated with sitxty six SSR markers covering
all the ten chromosomes. Out of these markers, twenty two markers were polymorphic while fourty four were
monomorphic. The size of the allele in polymorphic and monomorphic state represent the original molecular
character of the hybrid as well as their parental inbreds which helps in identification and distinction of hybrids and
its parental inbreds from other known varieties. Furthermore, DUS characterization can serve as additional
information.
Key words: Maize, Finger printing, Marker, Allele, Monomorpism, Polymorphism

Wild edible mushrooms (WEM) are known for its medicinal and nutritional value across the globe. WEM have become one of the most prized after food especially in the developed countries where people are mostly health conscious. The present study throws light on the diverse flora of WEMs of Nagaland and how it can be income generator for the tribal people with proper research in this aspect. Till now, the knowledge of distinguishing between edible and non-edible varieties is only confined to people who go for mushroom hunting. As such the indigenous knowledge remains with only those few people involved. The current data can pave the way for future research work and also make people aware of the many varieties of WEMs available in the state. A total of 33 WEMs were collected and identified during the peak mushroom season of the state i.e. from end May to September of every study year.

Content and purpose of study
An ethno medicinal survey of plants used by the  Idoma people of  Benue  State,
Nigeria, was carried out using a structural questionnaire via oral interview with
Traditional  Medicine  Practitioners  (TMPs)  and  knowledgeable  respondents  by
random  sampling  at  different  sites  within  the  Local  Government  Area.  Fifty
knowledgeable respondents were consulted for information on the plants used for
curative purposes.
Main findings
A  total  of  63  plants  belonging  to  36  families  were  identified.  Their  botanical
names,  ailments  treated,  plant  parts  used,  mode  of  administration  and  their
pharmaceutical  forms  were  provided.  Some  plants  like Azadirachta  indica,
Telferia  occidentalis  and Ocimum  gratissimum  are  used  for  the  treatment  of
common ailments such as malaria, anemia and stomach upset respectively among
the people.
Brief summary and potential implication
The need to document, establish the cultivation and sustainable harvesting as well
as  incorporation  of  traditional  medicinal  plants  into  the  primary  health  care
system in Nigeria were also highlighted. The data were analyzed in the forms of
‘specific  flora’  and  ‘general  floral’.  The  selectivity  of  a  plant  for  a  specific
ailment was done by comparing between the expected and observed values of the
proportion of citation of a plant for a specific disease. The difference (D) between
the  two  proportions  was  then  used  to  define  the  performance  index  (Ip),  which
ranged from 0 to 3.

We cordially announce the 21st Euro Biotechnology Congress to be held during October 11-12, 2018 Moscow, Russia. The attending delegates include Editorial Board Members of related international group journals. This is a fabulous open door for the representatives from universities and institutes to communicate with the world class scientists from all across the globe.

Abstract: Nanotechnology is considered to be a development way to gain efficient and potential approaches towards pest
control. The effect of both neem and peppermint oils as bulk, nano and loaded nano-emulsions have been studied against the
different biological aspects of the 2nd larval instar of Agrotis ipsilon (Hufn.) under laboratory conditions. Their effects on
enzymatic activities were investigated for the 2nd and 4th larval instars. The effects of adding the different formulations of either
neem or peppermint oils to artificial diet of 2nd instar larvae show significant elongation of the larval duration; percentage
mortalities were increased as well as larval malformations. It was recorded that nano-formulations (emulsion or loaded)
significantly decreased the pupal weight and significantly increased the pupal duration, percentage pupal mortalities and pupal
malformation. Adult longevity show insignificant effect, while female fecundity and percentage egg fertility show significant
decrease in comparison with the control. Results of enzymatic activities show marked effects of the three formulations of either
neem or peppermint oil. Significant inhibitions were recorded for amylase, invertase, trehalase, protease and alkaline
phosphatase. However, significant increase in the activities of cuticle phenoloxidase and chitinase were recorded. Marked
variations were recorded between nano-formulations and the bulk form.

With the adverse effects of insecticides used to control mosquitoes, development of an environment friendly, cost effective bio-control method is of utmost importance. Bacillus thuringiensis (Bt) provides an ideal solution which is quite effective against different kinds of mosquito larvae. The efficiency of using two local isolates namely Bt4 and Bt6e was evaluated and compared with standard Bt strain Bti HD-14. The effect of Bt strains against third instar larvae of Culex quinquefasciatus, Anopheles stephensi and Aedes aegypti was examined. Toxicity of Bti HD-14 was determined by obtaining mortality rate to find the effective concentrations against each mosquito type. A series of bioassays was carried out to determine mortality rate of three types of mosquitoes. Colony forming ability in LB medium was measured for each Bt strain to find out the growth pattern of two local strains and to compare with the standard. Toxicity level of LC 50 and LC 90 for Bti HD-14 against C. quinquefasciatus were 0.0026 and 0.0046 (mg/ml) respectively. There was no significant difference in mortality rate among three species of mosquito larvae within 24 hours of post treatment by Bt4 and Bt6e strains (p=0.01). Mortality rates of two local strains were similar though there was a difference between patterns of mortality rates during 9-18 hours after inoculation. Further, there was no significant difference between Bt standard strain and local strains in mortality rates (p=0.05). Based on the findings of the study it can be concluded that two local strains can be utilized as mosquito larvicides against the 3 rd instar larvae of tested mosquitoes.

Monoclonal antibodies (mAbs) are an effective tool in therapeutics and diagnostics. A novel approach called the single-cellRT-PCR-linked in vitro expression system (SICREX) enables the high-hroughput generation and screening of mAbs from single B cells. In this paper, instead of using Bcells, cDNAs were synthesized from single plasma cells of animmunized mouse spleen.The light chain (Lc) and the Fd portion of the heavy chain (Hc) genes of each cell were amplified separately and followed by overlapping PCR to add a T7 promoter, a ribosome-binding site, and a T7 terminator. The paired Lc and Hc genes were simultaneously expressed by an Escherichia coli in vitro transcription and translation system followed by ELISA to measure their affinity for the antigen. A Fab fragment with affinity against the antigen was obtained from plasma cells of an immunized mouse with hepatitis B surface antigen (HBsAg).

The potential role of PTI-1, in the natural story of prostate adenocarcinoma remains to be fully determined. PTI-1 expression was evaluated in human prostate cancer cell lines and in paraffin-embedded archive tissues. PTI-1 expression was found in Mycoplasma infected but not in non-infected cells. The lack of PTI-1 expression was also confirmed in fixed and paraffin-embedded human cancer prostate biopsies. The overall data indicate that, in prostate tumor cell lines, PTI-1 presence parallels Mycoplasma infection suggesting that PTI-1 might not necessarily play a major role in the onset of prostate tumorigenesis.

The Eukaryotic Translation Elongation Factors are a large protein family involved in the elongation step of eukaryotic translation but it has also various moonlight functions inside the cell both in normal and in pathological conditions. The proteins included in this family are EEF1A1, EEF1A2, EEF1B2, EEF1D, EEF1G, EEF1E1, enclosed their various isoforms, i.e. PTI-1, CCS-3, HD-CL-08, and MBI‐eEF1A. They are proteins all bound to cancer development and progression and show gene amplification, genomic rearrangements, and alteration of expression levels in many kinds of cancers. These abnormalities have undoubtedly repercussions on cellular biology and cellular behaviour in the various step of transformation and progression of cancer but surely should be considered also for the enhancement of invasiveness and for the metastasis. Thus, the Eukaryotic Translation Elongation Factors may possibly useful biomarkers for human cancers although more studies are needed to better elucidate their exact contribution as diagnostic, prognostic, and progression markers

Protein phosphatase 6 regulatory subunit 3 (PPP6R3) is a regulatory subunit of the PP6 holoenzyme complex involved in the turnover of serine and threonine phosphorylation events during mitosis. PPP6R3 shows abundant mRNA splicing variants and numerous functional protein isoforms. PPP6R3 gene is often involved in abnormal chromosomal translocations and it is found as a fusion gene partner in different kind of cancers.

Eukaryotic translation elongation factor 1 delta, alias EEF1D, is a protein-coding gene that plays a role in the elongation step of translation and considering its importance it is found frequently overexpressed in human cancer cells. This review collects the data on DNA/RNA, on the protein encoded and on the diseases where EEF1D is involved.

Eukaryotic translation elongation factor 1 beta 2, alias EEF1B2, is a protein-coding gene that plays a role in the elongation step of translation: In fact, it mediates GDP/GTP exchange on eEF1A. Considering its importance it is found frequently overexpressed in human cancer cells. This review collects the data on DNA/RNA, on the protein encoded and on the diseases where EEF1B2 is involved.

Presentazione dal titolo "Introduzione alla biologia molecolare di Mycoplasma sp." Contenuti: - Biologia molecolare dei micoplasmi - Cenni di epidemiologia - Meccanismo di infezione delle cellule - Principali patologie nell'uomo - Micoplasmi e trasformazione neoplastica - PTI-1 - Relazione tra PTI-1 e infezione da micoplasma [ENG] Presentation entitled "Introduction to the molecular biology of Mycoplasma sp." Contents: - Molecular biology of mycoplasmas - Outlines of epidemiology - Mechanism of cell infection - Main pathologies in humans - Mycoplasms and neoplastic transformation - PTI-1 - Relationship between PTI-1 and mycoplasma infection

The potential role of PTI-1, in the natural story of prostate adenocarcinoma remains to be fully determined. PTI-1 expression was evaluated in human prostate cancer cell lines and in paraffin-embedded archive tissues. PTI-1 expression was found in Mycoplasma infected but not in non-infected cells. The lack of PTI-1 expression was also confirmed in fixed and paraffin-embedded human cancer prostate biopsies. The overall data indicate that, in prostate tumor cell lines, PTI-1 presence parallels Mycoplasma infection suggesting that PTI-1 might not necessarily play a major role in the onset of prostate tumorigenesis.

Eukaryotic translation elongation factor 1 gamma, alias eEF1G, is a protein that plays a main function in the elongation step of translation process but also covers numerous moonlighting roles. Considering its importance in the cell it is found frequently overexpressed in human cancer cells and thus this review wants to collect the state of the art about EEF1G, with insights on DNA, RNA, protein encoded and the diseases where it is implicated.

The current study aimed to obtain staphylokinase (Sak) protein as a fibrinolytic agent from Escherichia coli as a nonpathogenic bacterium after transforming it with a cloned vector. From 630 clinical samples, 98 isolates of S. aureus were obtained and 81 (82.65%) were Sak producers. PCR amplification of the sak gene identified a single DNA band with the expected length of 408 bp in three S. aureus isolates. E. coli DH5 was successfully transformed with the sak gene. PCR amplification showed that the recombinant gene comprised 408 bp, which was the same size as the native gene amplified from S. aureus. Bacterial proteins of genetically engineered BL21(DE3) with cloned pET-28a-sak were separated according to their molecular weight and recombinant Sak appeared as a band with an estimated molecular weight ca 15.5 KDa. A non-induced sample with cloned vector was included as control. The fibrinolytic activity of recombinant Sak was higher than that of the native Sak. We found that the gene expression level and Sak production rate in the genetically engineered BL21(DE3) were similar to those of the native S. aureus.

Oocyte expressed protein, alias OOEP, is a component of the subcortical maternal complex (SCMC) that play its roles in oocytes and in early stages of embryogenesis. In this review it is done an insight on its DNA, its RNA, its protein encoded and on the diseases where OOEP is involved.

Genetic variation was surveyed at nine microsatellite loci and the mitochondrial control region (868 bp) to test for the presence of genetic stock structure in young-of-the-year Atlantic bluefin tuna (Thunnus thynnus thynnus) from the Mediterranean Sea. Bluefin tuna were sampled over a period of 5 years from the Balearic and Tyrrhenian seas in the western basin of the Mediterranean Sea, and from the southern Ionian Sea in the eastern basin of the Mediterranean Sea. Analyses of multilocus microsatellite genotypes and mitochondrial control region sequences revealed no significant heterogeneity among collections taken from the same location in different years; however, significant spatial genetic heterogeneity was observed across all samples for both microsatellite markers and mitochondrial control region sequences (FST = 0.0023, P = 0.038 and ΦST = 0.0233, P = 0.000, respectively). Significant genetic differentiation between the Tyrrhenian and Ionian collections was found for both microsatellite and mitochondrial markers (FST = 0.0087, P = 0.015 and ΦST = 0.0367, P = 0.030, respectively). These results suggest the possibility of a genetically discrete population in the eastern basin of the Mediterranean Sea.

BACKGROUND:

The human methionine synthase gene (MTR) is located on chromosome 1q43; it is of 105.24 kb and is made up of 33 exons. Methionine synthase is a cytoplasmic enzyme that requires methylcobalamin for activity and catalyzes the remethylation of homocysteine to methionine. In this reaction, the methyl group of 5-methyltetrahydrofolate is transferred to the enzyme bond cob(I) alamin to generate methylcobalamin, followed by the transfer of the methyl group to homocysteine to reform methionine.

MATERIALS AND METHODS:

The frequencies of the polymorphisms of MTR 2756A>G and MTR 2758C>G have been determined in this study in a sample of 491 individuals collected from all regions of Jordan and representing the Jordanian population. The different alleles and genotypes at the two polymorphic sites were identified using the Polymerase Chain Reaction - Restriction Fragment Length Polymorphism (PCR-RFLP) analysis.

RESULTS:

Showed that the percentages of the polymorphic alleles at the MTR 2756 position in the north, middle and south regions were 90.38, 92.65 and 83.69%, respectively, for the MTR 2756A allele, and were 9.61, 7.34 and 16.30%, respectively, for the MTR 2756G allele, with overall percentages in the whole Jordanian population of 90.73 and 9.27% for the MTR 2756A and MTR 2756G alleles, respectively. The percentages of the genotype MTR 2756AA were 82.90% in the northern region, 86.72% in the middle region and 71.73% in the southern region, and an overall percentage of MTR 2756AA in the whole Jordanian population was 83.50%. The frequencies of MTR 2756AG genotype in the northern, middle and southern regions were 14.95, 11.84 and 23.91%, respectively, with an overall percentage of 14.46% in the whole Jordanian population. The percentages of the genotype MTR 2756GG in the northern, middle and southern regions were 2.13, 1.42 and 4.34%, respectively, with an overall percentage of 2.04% in the whole Jordanian population. Only the wild type allele (C) of the MTR 2758C>G polymorphism was detected in this study. In addition, the association of MTR 2756A>G and MTR 2758C>G polymorphisms with the development of neural tube defects (NTDs) was examined using 17 cases of mothers from the northern part of Jordan, who gave birth to NTD affected children during the period of this study. Results showed no association between these two examined polymorphisms and the increase in maternal risk for giving birth to NTD children.

CONCLUSION:

results of this study recommend that examination should be done on larger populations to arrive at better conclusions. Also, more studies on gene–gene interaction should be done to examine the associations with NTDs.

Genet structure of an ectomycorrhizal (ECM) fungus Laccaria laccata was studied during a fruiting season and 9 months after the fruiting season in the volcanic desert on Mount Fuji. Below-ground samples of L. laccata were analyzed using SSR (microsatellite) markers. During fruiting season, most of the below-ground genets were identical to sporocarp genets of each sampling plot. In contrast, nine months after the sporocarp formation, many small genets were observed in each plot but previous year's sporocarp genet was not detected in 60% plots. Majority of new genets shared at least one common allele with the previous year's sporocarp in every SSR locus. This indicates, genets of L. laccata tend to disappear after sporocarp formation (within few months) and new offspring genets are generated from the spores disappeared from the sporocarp to the vicinity. The size and number of genet were significantly smaller and more in 'after fruiting season'. This result suggests, most of the newly generated genets disappear through competition with a sporocarp-producing genet that becomes dominant in the 'fruiting season.