Purple Top Research Articles

Unveiling the Significance of MRP 1 in Achieving Complete Remission Following Induction Therapy in Acute Myeloid Leukemia.

Increased expression of MRP 1 in AML patients results in the efflux of drugs from the cells, preventing the patient from achieving remission or potentially leading to relapse. Several studies have demonstrated that early identification of ABC transporter may yield favorable outcomes. The objectives of the study were to investigate the correlation between MRP 1 gene expression and MRP 1 protein levels and the response to remission induction in AML patients. A total of 40 AML patients were recruited from March 2021 to June 2022. Peripheral blood was collected in two tubes (yellow and purple top) to assess the MRP 1 gene and protein. For MRP 1 gene assessment, RNA was isolated from blood samples, cDNA was prepared, and qRT-PCR was performed to analyze gene expression. The relationship between the gene and complete remission was determined. Identification of MRP 1 protein was conducted using ELISA, and the relationship between protein levels and complete remission (CR) was explored. Most of the patients were aged between 25 and 39 years, encompassing both males and females. This study observed a clinical correlation between MRP 1 gene expression and complete remission. The findings revealed that 69.2 percent of patients with high gene expression failed to achieve complete remission, whereas the analysis of MRP 1 protein in relation to complete remission showed no statistical significance. The MRP1 gene showed high expression (66.7%) in patients with FLT3 mutation, whereas low expression of MRP1 was associated with a high occurrence (60%) of NMP1 mutation. Further comprehensive multicenter studies with larger sample sizes are required to validate the findings of this study. It is recommended to pinpoint the mechanism and regulation of MRP 1 and its interaction with other molecular pathways.

Zebra chip and purple top: An overview on two emerging diseases of potato

Zebra chip and purple top: An overview on two emerging diseases of potato

Differential Symptomology, Susceptibility, and Titer Dynamics Manifested by Phytoplasma-Infected Periwinkle and Tomato Plants.

Phytoplasmas are intracellular pathogenic bacteria that infect a wide range of plant species, including agriculturally important crops and ornamental trees. However, our understanding of the relationship between symptom severity, disease progression, and phytoplasma concentration remains limited due to the inability to inoculate phytoplasmas mechanically into new plant hosts. The present study investigated phytoplasma titer dynamics and symptom development in periwinkle and tomato, both infected with the same potato purple top (PPT) phytoplasma strain using a small seedling grafting approach. Virescence, phyllody, and witches'-broom (WB) symptoms sequentially developed in periwinkle, while in tomato plants, big bud (BB, a form of phyllody), cauliflower-like inflorescence (CLI), and WB appeared in order. Results from quantitative polymerase chain reaction (qPCR) targeting the PPT phytoplasma's 16S rRNA gene revealed that in both host species, phytoplasma titers differed significantly at different infection stages. Notably, the highest phytoplasma concentration in periwinkles was observed in samples displaying phyllody symptoms, whereas in tomatoes, the titer peaked at the BB stage. Western blot analysis, utilizing an antibody specific to PPT phytoplasma, confirmed substantial phytoplasma presence in samples displaying phyllody and BB symptoms, consistent with the qPCR results. These findings challenge the conventional understanding that phytoplasma infection dynamics result in a higher titer at later stages, such as WB (excessive vegetative growth), rather than in the early stage, such as phyllody (abnormal reproductive growth). Furthermore, the PPT phytoplasma titer was markedly higher in periwinkles than in tomato plants, indicating differing susceptibilities between the hosts. This study reveals distinct host responses to PPT phytoplasma infection, providing valuable insights into phytoplasma titer dynamics and symptom development, with implications for the future management of agricultural disease.

The Endoplasmic Reticulum Is a Key Battleground between Phytoplasma Aggression and Host Plant Defense.

Phytoplasmas are intracellular plant pathogens that heavily rely on host cell nutrients for survival and propagation due to their limited ability to synthesize essential substrates. The endoplasmic reticulum (ER), which plays a vital role in various cellular processes, including lipid and protein biosynthesis, is an attractive target for numerous intracellular pathogens to exploit. This study investigated the impact of potato purple top (PPT) phytoplasma infection on the ER in tomato plants. Abnormal accumulation of ER-resident proteins, disrupted ER network structures, and formation of protein aggregates in the phloem were observed using confocal microscopy and transmission electron microscopy, indicating a phytoplasma-infection-induced disturbance in ER homeostasis. The colocalization of phytoplasmas with the accumulated ER-resident proteins suggests an association between ER stress, unfolded protein response (UPR) induction, and phytoplasma infection and colonization, with the ER stress response likely contributing to the host plant's defense mechanisms. Quantitative real-time PCR revealed a negative correlation between ER stress/UPR activation and PPT phytoplasma titer, implying the involvement of UPR in curbing phytoplasma proliferation. Inducing ER stress and activating the UPR pathway effectively decreased phytoplasma titer, while suppressing the ER-resident protein, binding immunoglobulin protein (BiP) increased phytoplasma titer. These results highlight the ER as an intracellular battleground where phytoplasmas exploit host components for survival and multiplication, while host plants deploy defense mechanisms to counteract the invasion. Understanding the intricate interactions between phytoplasmas and plant hosts at the subcellular level, particularly within the ER, provides valuable insights for developing new strategies to control phytoplasma diseases.

Characterization of Pectobacterium Carotovoratum Subsp. Carotovorum causing Bacterial Soft Rot Disease of Turnip in Pakistan

This study aims to identify the Pectobacterium carotovorum subsp. carotovorum (Pcc) causing bacterial soft rot disease of turnip in Pakistan through conventional and molecular techniques. During the survey, four different bacterial isolates (VRSPCC01-04) were obtained from symptomatic turnip steckling (cv. purple top) at Vegetable Research Station Sahiwal, Punjab, Pakistan, and subjected for further analysis. All the isolates were characterized and identified based on morphological, biochemical, and molecular analysis (16S Ribosomal RNA). During morphological studies, the colony characters found deep cavities or pits formation in Crystal violet pectate medium (CVP), Creamy yellowish colony color was found on Yeast Extract Glucose calcium carbonate Medium (YGC), while creamy in color with the oval-shaped colony was observed on nutrient agar (NA) medium. In addition, the Pathogenicity test proved that all the tested isolates developed a typical water-soak symptom after 24 h and initiation of soft rot began the 48 h after inoculation on turnip roots. Results of biochemical tests showed that all isolates had a positive reaction for catalase, citrate, and an adverse reaction for Phosphatase and Indole test. The 16S rRNA sequences of isolates were 99-100% similar to the corresponding 16S rRNA sequence of the Pectobacterium carotovorum (PCC) reference isolates. Based on this, all identified bacterial isolates are first time reported as a causative agent of soft rot disease of turnip in Pakistan.

Efforts of researchers and other stakeholders to manage an unfolding epidemic: Lessons from potato purple top in Ecuador

Purple top is a phytosanitary problem affecting potato farmers in Ecuador. Since its emergence, stakeholders implement – with limited success – isolated efforts to reduce the spread of this problem. Hence, this study aimed to derive stakeholders’ enabling responses to manage the spread of emerging plant diseases and pests, and to analyse the spread of purple top in Ecuador together with the responses of the stakeholders. We review 108 articles on human, animal, and plant epidemics to derive enabling responses to manage the spread of emergent diseases and pests. Then, we describe the spread of purple top and stakeholders’ responses using an innovation history analysis approach. Our results show that managing epidemics calls for collaboration and coordination, communication and raising awareness, design and implementation of interventions, funding, supporting regulations and policies, research, and surveillance. We show that the complexity of the diagnosis of causal agent(s) of purple top caused uncertainty among stakeholders. Each actor experienced the purple top’s epidemic differently depending on her or his role. Poor coordination between scientists and other stakeholders, and lack of political lobby hindered the diagnosis and development of management strategies for purple top. A delayed identification, due to the complexity of the causal agent and poor coordination, can further hinder the mobilisation of necessary resources. Our results together indicate that managing epidemics requires more than identifying the causal agent(s) and vectors. We expect that the findings of this study are useful to improve preparedness for other plant disease epidemics affecting food security crops.

Nitrogen Nutrition Affect the Growth and Bulb Yield of Garlic (Allium Sativum L.)

Nitrogen (N) is one of the critical nutrients for productivity and quality of cultivated crop species. However, its overused in crop cultivation caused several environmental concerns. Hence its proper use in agriculture is not only pre-requisite for sustainable crop cultivation but also necessary to control environmental pollution. A field trial was designed at experimental area of Department of Horticulture, Sindh Agriculture university Tando Jam, Pakistanto test the response of garlic varietiesto N nutrition in terms of growth and bulb attributed traits during the winter season, 2020-21. The N effect on growth and yield associated traits of two garlic varieties (local white and purple top) were investigated by applying four Nrates that included 0, 50 kg, 100 kg and 150 kg ha-1. The findings of the trial exhibitedthat N application at various rates had substantial positiveimpact on the growth and bulb development of garlic at (P<0.05). The garlic plantation receivedNat maximum rate (150 kg ha-1) resulted tallest plants,better neck thicknesswith more leaves plant-1. The N also positively affected the bulb yield and other yield parameters of garlic.The maximum bulb yield was obtained when plants were nourished with 150 Kg N ha-1that was 381.98 % more than the control. It was noted that increase inN rates was statistically dominant over other doses. The varietal influence on garlic indicated that variety purple top performed better than local white in terms of yield associated traits..These findings provide new insight regarding the significance of N nutrition in improving the growth and bulb yield of garlic genotypes

Cas12a-Based Diagnostics for Potato Purple Top Disease Complex Associated with Infection by ‘Candidatus Phytoplasma trifolii’-Related Strains

'Candidatus Phytoplasma trifolii' is a cell wall-less phytopathogenic bacterium that infects many agriculturally important plant species such as alfalfa, clover, eggplant, pepper, potato, and tomato. The phytoplasma is responsible for repeated outbreaks of potato purple top (PPT) and potato witches' broom (PWB) that occurred along the Pacific Coast of the United States since 2002, inflicting significant economic losses. To effectively manage these phytoplasmal diseases, it is important to develop diagnostic tools for specific, sensitive, and rapid detection of the pathogens. Here we report the development of a DNA endonuclease targeted CRISPR trans reporter (DETECTR) assay that couples isothermal amplification and Cas12a transcleavage of fluorescent oligonucleotide reporter for highly sensitive and specific detection of 'Candidatus Phytoplasma trifolii'-related strains responsible for PPT and PWB. The DETECTR assay was capable of specifically detecting the 16S-23S ribosomal DNA intergenic transcribed spacer sequences from PPT- and PWB-diseased samples at the attomolar sensitivity level. Furthermore, the DETECTR strategy allows flexibility to capture assay outputs with fluorescent microplate readers or lateral flow assays for potentially high-throughput and/or field-deployable disease diagnostics.

Breeding value of leafy and root turnip samples from the VIR collection

Turnip, Brassica rapa L. subsp. rapa is the oldest vegetable crop in Russia. Turnip varieties are fastgrowing, productive, have valuable biochemical properties, cold-resistant, relatively easy to grow. To expand the vegetable assortment, new varieties and hybrids should be created, first of all, the types of varieties that are not in the State Register of Russia. The purpose of the work: using the data of a long-term study of the VIR collection to identify valuable material for breeding.Methods. Field study of 170 samples of the VIR collection was carried out in the NPB "Pushkin and Pavlovsk laboratories of VIR" (Leningrad region) in 2001-2020. Sowing was carried out on June 26- 28. Each sample was sown on a single-row plot 2 m long in three repetitions. As a control, turnip varieties included in the State Register were used – the variety of leaf turnip Sapphire, the table turnip variety Petrovskaya-1 and the fodder turnip variety Osterzundomsky. The description was carried out according to the Methodological Guidelines for the study and maintenance of the VIR world collection, with author's additions.Results. The best leaf turnip samples, found during the study has a large plant mass (up to 1.42±0.32 kg in the Early top F1 sample (k-366, Japan), many samples exceeded the Sapphire standard for this trait. Some of the samples of table turnips also exceed the Petrovskaya-1 standard by weight of the root (Purple top (k-1345, Nepal), Pekki (vr.2176, Finland)). Samples Solovetskaya (k-803, Russia, Murmansk region), Gul Findlandsk (vr.2175, Norway) have a bright yellow flash of the root, are easy to store and resistant to damage by cabbage flies. Among the turnip samples distinguished by yield (Table 5), the most interesting are the turnips Ova Dæhnfeldt (vr.2183, Denmark) and Korova (vr.2187, Sweden), which have yellow flesh and an increased content of β-carotene in roots.Conclusion. It was possible to identify new samples belonging to groups of leaf and root turnips, that are not represented on the seed and food market of Russia. Such samples belong to morphologically different forms, are productive, carry a complex of valuable economic and biochemical characteristics and are perspective material for inclusion in programs for the selection of leaf and root crops.

Molecular biological study on the survival strategy of phytoplasma

Phytoplasmas (genus ‘Candidatus Phytoplasma’) are plant pathogenic bacteria that reside intracellularly within the plant phloem. They infect hundreds of plant species worldwide and induce a wide range of symptoms, including stunting, yellowing, witches’ broom, phyllody, virescence, proliferation, and purple top. The phytoplasma genome has unique features that overturn commonly known wisdom. For example, ATP synthase, which is thought to be essential for life, is not encoded, which provides opportunities for reconsidering the question what is life? Phytoplasmas cause dramatic changes in plant development and parasitizes not only plants but also vector insects by host switching. Therefore, the molecular mechanisms of how phytoplasmas manipulate host plants and how they infect different hosts are of particular interest. This review summarizes the history of phytoplasma discovery, unique survival strategies revealed by genome sequencing, and the molecular mechanisms by which phytoplasmas manipulate host plants.

Molecular detection of phytoplasmas in potato psyllids in ecuador

Since the potato purple top appeared and the potato psyllid was reported in Ecuador, crop losses have been significant. So far, phytoplasmas enclosed in two ribosomal groups (16SrI-F, 16SrII) and Candidatus Liberibacter solanacearum’ were molecularly identified in symptomatic potato plants. Later, Bactericera cockerelli, a potato psyllid, was also first reported and correlated to the potato purple top disease. Molecular identification of the pathogens involved in this disease has been quite elusive. Samples of potato psyllids were analyzed in this study resulting in the identification of phytoplasmas enclosed in group 16SrXI, and others related to Candidatus Phytoplasma prunorum’ and Ca. P. solani’. Potato purple top in Ecuador is a complex disease that constitutes a severe threat for the potato production in the region.

Thiamin stimulates growth and secondary metabolites in turnip (Brassica rapa L.) leaf and root under drought stress

Thiamin, an important member of the vitamin B family, is believed to play a significant role in mitigating environmental stresses including drought stress. In turnip, drought stress causes a reduced growth, biomass yield, pigment content, total phenolics and ascorbic acid (AsA), particularly at 50% field capacity (F.C.) in the two cultivars (cv) studied. However, a significant enhancement was observed in the contents of leaf proline, glycinebetaine (GB), malondialdehyde (MDA), hydrogen peroxide (H2 O2 ) and the activities of catalase (CAT) and superoxide dismutase (SOD) as well as root proline, GB, total phenolics, AsA, H2 O2 , MDA and the activities of peroxidase (POD) and SOD. However, foliar-applied thiamin significantly improved (particularly 100 mM) all the growth attributes, photosynthetic pigments, leaf and root osmoprotectants (GB and proline), AsA, total phenolics and the activities of enzymatic antioxidants such as SOD and POD as well as root CAT in both turnip cultivars under drought stress conditions. Foliar application of thiamin was effective in decreasing the leaf and root H2 O2 and MDA content in both cultivars particularly at 50% F.C. Thiamin-induced growth of both turnip cultivars, particularly of cv. Purple Top, was found to be associated with increased photosynthetic pigments, proline and GB contents and antioxidant capacity, but reduced levels of reactive oxygen species (ROS) under water deficit conditions. So, it is suggested that exogenous application of thiamin can be effective in improving drought tolerance of plants.

Ixeris Chinensis is a New Host for Peanut Witches' Broom Phytoplasma, a 16SrII-V Subgroup Strain in Taiwan.

Ixeris chinensis (Thunb. ex Thunb.) Nakai, a perennial herbaceous plant that belongs to the family of Asteraceae, is widely distributed at mid-low altitude regions in Taiwan. I. chinensis is commonly used as traditional herbal medicine for the treatment of inflammation, bronchitis, pneumonia, and diarrhea. In March 2020, disease symptoms such as shoot proliferation, phyllody, virescence, purple top, and witches' broom were observed on I. chinensis at the sansheng community park in Mailiao, Yunlin County, Taiwan. Totally, eight I. chinensis plants were checked and half of them were symptomatic. These disease symptoms are similar to those associated with peanut witches' broom (PnWB) disease identified in the same area (Liu et al. 2015). Three samples mixed with leaf, stem, and flower were tested including one healthy and two symptomatic I. chinensis. The total DNA of each sample was extracted and examined by nested PCR for the amplification of 16S rDNA with the phytoplasma universal primer pairs P1/P7 followed by R16F2n/R16R2 (Lee et al. 1993). A specific signal of expected size (1.2 kb) for 16S rDNA was only detected in the symptomatic I. chinensis, but not in healthy I. chinensis. The nucleotide sequence (accession no. MT416114) of the amplified DNA fragment using primer pairs P1/P7 from symptomatic I. chinensis is identical to that of GenBank accession NZ_AMWZ01000008 (complement [31109 to 32640]) of phytoplasma associated with PnWB disease (Chung et al. 2013). Analysis of the virtual RFLP pattern of MT416114 generated by iPhyClassifier revealed that the phytoplasma detected in symptomatic I. chinensis belongs to a 16SrII-V subgroup. The total protein of each sample was also extracted and examined by western blotting using the polyclonal antibody raised against Imp protein of purple coneflower witches' broom phytoplasma (Chien et al. 2020), which is identical with that (accession no. ADD59806) of PnWB phytoplasma. An expected signal of 19 kDa specific for Imp was detected in symptomatic I. chinensis, but not in healthy I. chinensis. Subsequent PCR, DNA sequencing and western blotting assays further confirmed that the gene encoding a SAP11-like protein was only detected in symptomatic I. chinensis, and shares 100% identity with that (accession no. EMR14684) of PnWB phytoplasma. Our results indicate that PnWB phytoplasma causes disease in I. chinensis, a common weed, which may act as an alternative natural host and facilitate the spreading of phytoplasma disease in Taiwan.

Virulence/avirulence patterns among Leptosphaeria maculans isolates determines expression of resistance, senescence and yellowing in cotyledons of Brassica napus

Blackleg, (stem canker, Leptosphaeria maculans), is the most economically important disease on oilseed rape (Brassica napus). Studies were undertaken to determine the effect of three different L. maculans isolates with different virulence/avirulence patterns on the expression of qualitative resistance and senescence on cotyledons of 11 B. napus genotypes. There were significant differences between genotypes (P Scotia > Amber x Commanche DH Line. Westar, Drummonds Purple Top, and Parkside were the most susceptible. L. maculans isolates D5 and D13 were the most pathogenic, showing the greatest disease index and lesion diameters. Genotypes such as LSF0530, Scotia, Duell and Barrel responded with a distinct hypersensitive response, whereas Parkside, Drummonds Purple Top, Amber x Commanche DH Line and Westar all showed a distinct yellow halo around the lesions, indicative of cotyledon senescence likely involving programmed cell death (PCD). However, expression of the distinct yellow halo symptom was not observed in all B. napus genotypes infected by one specific L. maculans isolate nor in one specific B. napus genotype challenged with all the isolates. We believe that this is the first report, i), showing the involvement of a distinct yellow halo (senescence/PCD) associated with both ‘typical’ lesions and with an expression of HR in cotyledons of winter-type B. napus genotypes challenged with L. maculans isolates and, ii), that expression of the distinct yellow halo is clearly dependent on the interaction between particular isolate virulence(s) x particular winter-type B. napus genotype combinations.

Uso de plantas medicinais na Umbanda e Candomblé em associação cultural no município de Puxinanã, Paraíba

A utilização das plantas medicinais em práticas ritualísticas é algo bastante comum nas religiões existentes no Brasil, diante disso, o presente estudo apresenta uma pesquisa sobre o uso das plantas medicinais na Associação Cultural de Umbanda e Candomblé localizada em Puxinanã, Paraíba. Foram realizadas 50 entrevistas com pessoas que frequentam o terreiro, no qual mostra o uso intenso das ervas naturais nos rituais, sendo as três mais citadas: a jurema preta (Mimosa tenuiflora), liamba (Vitex agnus castus L.) e Pião roxo (Jatropha gossypiifolia L.), constatou-se também que todos os entrevistados acham importante a utilização das ervas, evidenciando a necessidade das mesmas se fazerem presentes nas ritualísticas da Umbanda e Candomblé. Os resultados confirmam que os frequentadores de terreiro possuem um grande conhecimento e fazem uso nos rituais com as plantas medicinais, preservando a cultura popular na religião.

Quantitative Analysis of Lactate and Other Organic Acids in Plasma: Preanalytical Challenges

Abstract Background and Objectives We developed a gas chromatography–mass spectrometry (GC-MS) method for the analysis of organic acids (OAs) in plasma using liquid-liquid extraction with acidified methanol. This method allows for the simultaneous analysis of several metabolites associated with energy metabolism, including lactic and pyruvic acids. Here, we evaluated the effect of different preanalytical variables (anticoagulants, delayed plasma separation, fasting vs nonfasting status) on plasma organic acid quantitation. Methods Nine organic acids (lactic, pyruvic, 3-hydroxybutyric, acetoacetic, 2-ketoisovaleric, 2-keto-3-methylvaleric, 2-ketoisocaproic, succinic, and glutaric) were extracted by deproteinizing with acidified methanol, oximated to preserve ketoacids, converted to volatile trimethylsilyl derivatives, and detected by GC-MS. Compound identification was obtained by retention time and fragmentation spectra using Agilent MassHunter software. We performed several comparison studies using blood collected from self-reported healthy participants (24-60 years of age; 14 females, 13 males). The effect of fasting/nonfasting status was evaluated in 14 participants by comparing samples collected after overnight fasting and 4 hours after meal. Paired t test was used for the statistical analysis of results. Results When compared to the preferred sample type, sodium heparin (green top) plasma EDTA plasma (purple top, n = 9) and serum (red top or serum separator tube; n = 5) displayed an unacceptable variability for several analytes. The biggest differences were observed for lactic and pyruvic acids. Lactic acid was significantly lower in EDTA plasma (9%-35% decrease; P = .0001), but higher in serum (4%-63% increase; P = .014), while pyruvic acid was significantly higher in EDTA plasma (4%-145% increase; P = .0002). The measurements of lactic and pyruvic acids were also affected by delayed separation of plasma in samples kept at room temperature. We observed a marked increase in lactate and decrease in pyruvate levels (+42% and –43% mean change, respectively) in plasma separated from whole blood after 2 hours of collection compared to samples processed within 30 minutes. Plasma separated 1 hour postcollection was acceptable. Overnight fasting (12-14 hours) did not have a significant impact on ketones (3-hydroxybutyric and acetoacetic acids) or branched-chain ketoacids (2-keto-3-methylvaleric, 2-ketoisocaproic, 2-ketoisovaleric acids). In contrast, fasting samples displayed higher lactic acid concentrations compared to nonfasting samples (24% increase on average, P = .0072), although in most fasting samples (13 out of 14), lactate concentrations were still within established normal range (600-2,600 µmol/L). Conclusions We identified several preanalytical factors influencing plasma organic acid quantitation, particularly lactic and pyruvic acids. Since those are critical analytes, suggestive of primary or secondary metabolic acidosis, appropriate steps should be taken to ensure sample quality during collection and processing.

Effect of time and temperature on stability of progestagens, testosterone and cortisol in Asian elephant blood stored with and without anticoagulant.

The value of biological samples collected in the field is compromised if storage conditions result in analyte degradation, especially in warmer climates like Thailand. We evaluated the effects of time and temperature on immunoactive steroid hormone stability in Asian elephant (Elephas maximus) blood stored with and without an anti-coagulant before centrifugation. For each elephant (5 male, 5 female), whole blood was aliquoted (n = 2ml each) into 13 red top (without anticoagulant) or purple top (with anticoagulant) tubes. One tube from each treatment was centrifuged immediately and the serum or plasma frozen at -20°C (Time 0, T0). The remaining 12 aliquots were divided into stored temperature groups: 4°C, room temperature (RT, ~22°C), and 37°C, and centrifuged after 6, 24, 48 and 62h of storage. Serum and plasma concentrations of progestagens in females, testosterone in males and cortisol in both sexes were quantified by validated enzyme immunoassays. Steroid concentration differences from T0 were determined by a randomized complete block ANOVA and Dunnett's tests. The only evidence of hormone degradation was cortisol and testosterone concentrations in serum stored at 37°C. Testosterone concentrations declined by 34% at 48h and 52% at 62h, cortisol was decreased by 19% after 48h and 27% after 62h at 37°C, respectively. None of the other aliquots displayed significant changes over time at any temperature. In conclusion, steroids appear to be stable in blood for nearly 3days at room or refrigeration temperatures before centrifugation; steroids in samples with ethylenediaminetetraacetic acid were particularly stable. However, warmer temperatures may negatively affect steroids stored without anti-coagulant, perhaps due to red blood cell metabolism. Thus, under field conditions with no access to cold or freezer temperatures, collection of plasma is a better choice for elephants up to at least 62h before centrifugation.

Detection and molecular characterization of a 16SrI-F phytoplasma in potato showing purple top disease in Ecuador

Potato purple top (PPT) disease has been reported worldwide, including Canada, United States, Mexico, Russia and Pakistan. During surveys carried out in a recent epidemic in Ecuador of potato purple top disease phytoplasmas were identified in symptomatic potatoes as well as in some tissue cultured potato shoots by PCR/RFLP analyses and by sequencing and phylogenetic clustering. The majority of the samples resulted positive to phytoplasmas enclosed in the ribosomal subgroup 16SrI-F, for the first time detected in potato and in the American continent. Further studies will allow verification of the transmission and further presence in Ecuador of this phytoplasma associated with purple top disease of potatoes.

Childhood Leukemias in Khyber Pakhtunkhwa and Afghan Children Visiting to Hayatabad Medical Complex Hospital

Background: Khyber Pakhtoonkhwa and Afghanistan located in South Central Asia which is most prevalent area for childhood leukemia. In this region, no previous data or study was available on estimated cases per year of different types of childhood leukemia. The aim of the present study is to evaluate the frequency of different types of childhood leukemia in the children from different districts of Khyber Pakhtoonkhwa (KP) and Afghanistan presenting to Hayat Abad Medical Complex Hospital, Peshawar. Methods: This descriptive cross-sectional study was conducted in pathology department Hayat Abad Medical Complex hospital, Peshawar, Pakistan. Duration of the present study was, from January 2014 to December 2016. A total number of 605 children were enrolled up to 15 years of age, who were suspected to have leukemia, went through bone marrow examination. 3 ml blood was collected in EDTA tube (purple top) and complete blood count was prepared by hematology analyzer (Ruby cell dyne, Abbott, USA). By aseptic techniques bone marrow aspiration and bone marrow trephine biopsy samples were collected from all patients. Slides were papered from Bone marrow aspirates, fixed with methanol and stained with Giemsa, myeloperoxidase and periodic acid Schiff stain. Trephine biopsy slides were stained by Hematoxylene and Eosin and Reticulin stain. Immunohistochemistry was done after initially examining bone marrow aspirate slides. All data was documented and statistical analysis was performed by SPSS-20 software. Findings: Among 605 children suspected to have leukemia, 281 (46.5%) children were diagnosed with different types of childhood leukemia. Out of 281 diagnosed patients, 231(82.2%) were Pakistani children and rest of the 50 (17.8%) were Afghani children. The studied population consist of 173(61.6%) males and 108(38.4) females. Their age ranged from 3 months to 15 years with median age of 9.8 years. Out of 281 cases, 210 (74.73%) were diagnosed to have acute lymphoblastic leukemia and rest of the children were 62 (22.06%) acute myeloid leukemia, 7 (2.44%) chronic myeloid leukemia, 2 (0.71%) had Juvenile Chronic Myelomonocytic Leukemia (JMML) and 1 (0.35%) child was report with Chronic Myelomonocytic leukemia (CMML). There was no significant differences seen in prevalence among Afghan children and Pakistani children except ALL, which is more frequent in Afghan children compared to Pakistani children and no single case of CMML, were diagnosed in Afghan children. Conclusion: In the present study, acute lymphoblastic leukemia is most frequent leukemia in the children of Khyber Pakhtoonkhwa and Afghanistan. Juvenile chronic Myelomonocytic leukemia is found the less common leukemia in the present study.

Molecular Characterization of the Vitellogenin Receptor inBactericera cockerelli(Sulc)

Abstract. Potato purple top and zebra chip diseases are transmitted by potato psyllid, Bactericera cockerelli (Sulc), to tomato, Solanum lycopersicum L., and potato, Solanum tuberosum L., crops. The aim of this study was to characterize and quantify vitellogenin receptor in potato psyllid. We obtained partial characterization of the receptor by end-point PCR and sequencing using capillary electrophoresis. Relative quantification by real-time PCR was used to determine expression levels of the receptor in females and males. Similarity in vitellogenin sequence obtained in a laboratory was 89% compared with the sequence of the entire transcriptome of the species reported in GenBank. Expression analysis showed the vitellogenin receptor significantly expressed in potato psyllid females compared with males from potato plants. Further study of the vitellogenin receptor of potato psyllid is needed to determine its complete sequence and potential target to control the pest.