Genomic characterization of Puccinia triticina using molecular marker technology / Caracterização genômica de Puccinia triticina usando tecnologia de marcadores moleculares

Leaf rust, caused by Puccinia triticina, is the most common rust disease of wheat. The fungus is an obligate parasite capable of producing infectious urediniospores. To study the genetic structure of the leaf rust population 20 RAPD primers were evaluated on 15 isolates samples collected in Pakistan. A total of 105 RAPD fragments were amplified with an average of 7 fragments per primer. The number of amplified fragments varied from 1 to 12. GL Decamer L-07 and GL Decamer L-01 amplified the highest number of bands (twelve) and primer GL Decamer A-03 amplified the lowest number of bands i.e one. Results showed that almost all investigated isolates were genetically different that confirms high genetic diversity within the leaf rust population. Rust spores can follow the migration pattern in short and long distances to neighbor areas. Results indicated that the greatest variability was revealed by 74.9% of genetic differentiation within leaf rust populations. These results suggested that each population was not completely identical and high gene flow has occurred among the leaf rust population of different areas. The highest differentiation and genetic distance among the Pakistani leaf rust populations were detected between the leaf rust population in NARC isolate (NARC-4) and AARI-11and the highest similarity was observed between NARC isolates (NARC-4) and (NARC-5). The present study showed the leaf rust population in Pakistan is highly dynamic and variable.

A ferrugem da folha, causada por Puccinia triticina, é a ferrugem mais comum do trigo. O fungo é um parasita obrigatório, capaz de produzir urediniósporos infecciosos. Para estudar a estrutura genética da população de ferrugem da folha, 20 primers RAPD foram avaliados em 15 amostras de isolados coletadas no Paquistão. Um total de 105 fragmentos RAPD foram amplificados com uma média de 7 fragmentos por primer. O número de fragmentos amplificados variou de 1 a 12. GL Decamer L-07 e GL Decamer L-01 amplificaram o maior número de bandas (doze), e o primer GL Decamer A-03 amplificou o menor número de bandas, ou seja, um. Os resultados mostraram que quase todos os isolados investigados eram geneticamente diferentes, o que confirma a alta diversidade genética na população de ferrugem da folha. Os esporos de ferrugem podem seguir o padrão de migração em distâncias curtas e longas para áreas vizinhas. Os resultados indicaram que a maior variabilidade foi revelada por 74,9% da diferenciação genética nas populações de ferrugem. Esses resultados sugeriram que cada população não era completamente idêntica e um alto fluxo gênico ocorreu entre a população de ferrugem da folha de diferentes áreas. A maior diferenciação e distância genética entre as populações de ferrugem da folha do Paquistão foram detectadas entre a população de ferrugem da folha no isolado NARC (NARC-4) e AARI-11 e a maior similaridade foi observada entre os isolados NARC (NARC-4) e (NARC-5). O presente estudo mostrou que a população de ferrugem da folha no Paquistão é altamente dinâmica e variável.

Physical, Psychological and Social Impact of COVID-19 Pandemic on Healthcare Workers at a COVID Designated Bangladeshi Public Hospital.

The world has been devastated facing the outbreak of a novel infectious disease known as Corona virus disease (COVID-19). This has been declared as a pandemic by the World Health Organization. The frontline health care workers, who are directly involved in the diagnosis, treatment and care of patients with COVID-19, are taking significant personal risks on their own health and those of their family members. Objectives of the study include establishing the physical, psychological and social impact experience by the healthcare workers serving in public hospitals of Bangladesh. This prospective cross-sectional observational study was carried out at Kuwait Bangladesh Friendship Government Hospital, the first Covid-19 designated hospital of Bangladesh between the 1st June and the 31st August, 2020. A total of 294 doctors, nurses, ward boys and ailed healthcare workers were included in this study via purposive sampling. The study found statistically significant (p value 0.024) difference of medical co-morbidities between Covid-19 positive and Covid-19 negative groups of health care professionals. Significant association was found between duration of work and presence during aerosol generating procedure with COVID infectivity of the study subjects. 72.8% respondents experienced public fear of contracting the virus from them and 69.0% noticed negative attitude of the society towards them. Eighty five percent (85.0%) did not get any community support during this pandemic crisis. The health care professionals engaged in COVID-19 treatment have been taking significant personal risk on their life in terms of physical, psychological and social perspective. Providing safeguard to the health care workers are integral components of public health measures for addressing the COVID-19 pandemic. Special interventions to promote their physical wellbeing and arrangement of adequate psychological training need to be immediately implemented to cope up this critical situation.

Vigilancia y manejo en consulta de los tumores de vejiga Ta de bajo grado / Surveillance and office management of low-grade Ta bladder tumors

Objetivo: Los pacientes con cáncer de vejiga (CV) Ta, de grado bajo (GB) 1-2, presentarán con frecuencia una «recidiva», aunque rara vez una progresión a un estadio más avanzado. Si bien las directrices actuales mencionan la vigilancia y el manejo en consulta para estos tumores nuevos o recurrentes, la resección transuretral es el tratamiento realizado con mayor frecuencia. El objetivo de este estudio es determinar la seguridad de la vigilancia y/o la cauterización ambulatoria. Materiales y métodos: Este estudio se llevó a cabo como un análisis retrospectivo de una serie de casos de 45 pacientes que tenían CV recurrente con apariencia de GB Ta y que fueron tratados principalmente con vigilancia y/o cauterización en la consulta. Se excluyeron los pacientes con carcinoma in situ. La variable de valoración principal fue la progresión a un estadio más avanzado. Resultados: La mediana de seguimiento fue de 62 meses. En 41 (91%) pacientes no hubo progresión del estadio. Tres pacientes recidivaron con CV grado alto (GA) T1; uno de ellos está recibiendo inmunoterapia sistémica. Un paciente desarrolló un CV GA T2 y fue tratado con un protocolo preservador de la vejiga. A 40 (89%) pacientes se les realizó cauterización en consulta. Once recibieron BCG y 26 recibieron quimioterapia intravesical tras la cauterización. Cinco (11%) pacientes desarrollaron CV GA durante el seguimiento. Ningún paciente falleció. Ninguno de los 17 (38%) pacientes hispanos tuvo progresión.ConclusionesLa vigilancia activa y/o la cauterización ambulatoria para pacientes con tumores vesicales pequeños GB Ta es segura, reduce costes y mejora la calidad de vida al evitar las resecciones transuretrales. (AU)

Objective: Patients with low-grade (LG), grade 1-2, Ta bladder cancer (BC) will frequently have a “recurrence”. However, they rarely progress in stage. Although current guidelines mention surveillance and office management for these new or recurrent tumors, transurethral resection is the most common treatment. The purpose of this study is to determine if surveillance and/or office cautery is safe. Materials and methods: This study was conducted as a retrospective case series analysis of 45 patients who had recurrent LG Ta appearing BC and were managed primarily with surveillance and/or office cautery. Patients with carcinoma in-situ were excluded. The primary outcome was stage progression. Results: Median follow-up was 62 months. Forty-one (91%) patients did not progress in stage. Three patients recurred with high grade (HG) T1 BC; one is receiving systemic immunotherapy. One patient developed HG T2 BC and was treated with a bladder preservation protocol. Forty (89%) patients underwent office cauterization. Eleven received BCG and 26 received post-cautery intravesical chemotherapy. Five (11%) patients developed HG BC during follow-up. No patients died. None of the 17 (38%) Hispanic patients had progression. Conclusions: Active surveillance and/or office cautery for patients with small recurrent LG Ta bladder tumors is safe, reduces cost and improves quality of life by avoiding transurethral resections. (AU)

Surveillance and office management of low-grade Ta bladder tumors.

OBJECTIVE: Patients with low-grade (LG), grade 1-2, Ta bladder cancer (BC) will frequently have a "recurrence". However, they rarely progress in stage. Although current guidelines mention surveillance and office management for these new or recurrent tumors, transurethral resection (TURBT) is the most common treatment. The purpose of this study is to determine if surveillance and/or office cautery is safe. MATERIALS AND METHODS: This study was conducted as a retrospective case series analysis of 45 patients who had recurrent LG Ta appearing bladder cancer (BC) and were managed primarily with surveillance and/or office cautery. Patients with carcinoma in-situ were excluded. The primary outcome was stage progression. RESULTS: Median follow up was 62 months. 41 (91%) patients did not progress in stage. Three patients recurred with HG T1 BC; one is receiving systemic immunotherapy. One patient developed HG T2 BC and was treated with a bladder preservation protocol. 40 (89%) patients underwent office cauterization. Eleven received BCG and 26 received post-cautery intravesical chemotherapy. Five (11%) patients developed HG BC during follow up. No patients died. None of the 17 (38%) Hispanic patients had progression. CONCLUSIONS: Active surveillance and/or office cautery for patients with small recurrent LG Ta bladder tumors is safe, reduces cost and improves quality of life by avoiding TURBTs.

Genomic characterization of Puccinia triticina using molecular marker technology.

Leaf rust, caused by Puccinia triticina, is the most common rust disease of wheat. The fungus is an obligate parasite capable of producing infectious urediniospores. To study the genetic structure of the leaf rust population 20 RAPD primers were evaluated on 15 isolates samples collected in Pakistan. A total of 105 RAPD fragments were amplified with an average of 7 fragments per primer. The number of amplified fragments varied from 1 to 12. GL Decamer L-07 and GL Decamer L-01 amplified the highest number of bands (twelve) and primer GL Decamer A-03 amplified the lowest number of bands i.e one. Results showed that almost all investigated isolates were genetically different that confirms high genetic diversity within the leaf rust population. Rust spores can follow the migration pattern in short and long distances to neighbor areas. Results indicated that the greatest variability was revealed by 74.9% of genetic differentiation within leaf rust populations. These results suggested that each population was not completely identical and high gene flow has occurred among the leaf rust population of different areas. The highest differentiation and genetic distance among the Pakistani leaf rust populations were detected between the leaf rust population in NARC isolate (NARC-4) and AARI-11and the highest similarity was observed between NARC isolates (NARC-4) and (NARC-5). The present study showed the leaf rust population in Pakistan is highly dynamic and variable.

The impact of fluid optimisation before induction of anaesthesia on hypotension after induction.

Intra-operative hypotension is a known predictor of adverse events and poor outcomes following major surgery. Hypotension often occurs on induction of anaesthesia, typically attributed to hypovolaemia and the haemodynamic effects of anaesthetic agents. We assessed the efficacy of fluid optimisation for reducing the incidence of hypotension on induction of anaesthesia. This prospective trial enrolled 283 patients undergoing radical cystectomy and randomly allocated them to goal-directed fluid therapy (n = 142) or standard fluid therapy (n = 141). Goal-directed fluid therapy patients received fluid optimisation based on stroke volume response to passive leg raise before induction; those with positive passive leg raise received intravenous crystalloid fluid boluses until stroke volume was optimised. Baseline mean arterial pressure was measured on the morning of surgery and on arriving in the operating theatre. This post-hoc analysis defined haemodynamic instability as either a > 30% relative drop in mean arterial pressure compared with baseline or absolute mean arterial pressure < 55 mmHg, within 15 min of induction. Forty-two (30%) goal-directed fluid therapy patients underwent fluid optimisation after finding an intravascular fluid deficit via passive leg raise testing; 106 (75%) goal-directed fluid therapy and 112 (79%) standard fluid therapy patients met criteria for haemodynamic instability. There was no significant difference in the incidence of haemodynamic instability between the goal-directed fluid therapy and standard fluid therapy groups using absolute mean arterial pressure drop below 55 mmHg (p = 0.58) or using pre-surgical testing or pre-surgical mean arterial pressure values as baseline (p = 0.21, p = 0.89, respectively); however, the difference in the incidence of haemodynamic instability was significant using the operating theatre baseline mean arterial pressure (p = 0.004). We conclude that fluid optimisation before induction of general anaesthesia did not significantly impact haemodynamic instability.

IDENTIFICATION OF QTLs ON CHROMOSOME 1B FOR GRAIN QUALITY TRAITS IN BREAD WHEAT (TRITICUM AESTIVUM L.).

The present study was designed considering the importance of grain quality traits, genetic diversity and marker-trait association analysis in wheat. A significant amount of genetic diversity was found for various seed traits though the genotypes included in the study were found structured. The extent of polymorphism was high with a range of 2-13 alleles and average of 6.5 alleles per locus. Population structure was detected with 30 unlinked SSRs that divided the population of 92 genotypes in three sub-populations. Extensive LD extent was found on chromosome 1B with 42 SSRs specific for 1B chromosome. Marker-trait associations were determined using mixed linear model, where, population structure and kinship calculated on the basis of unlinked markers were covariated with 1B specific markers and traits data. Eight QTLs for five traits including protein, gluten contents, test weight bread and chapati making quality. Protein content, test weight, bread quality and Glu-B1 were found significantly associated with primers WMC419 (32 cM); WMC128 (30 cM), WMC419 (32 cM); WMC818 (17 cM) and WMC416 (44 cM), respectively.

Cost trajectories for cancer patients.

BACKGROUND: Health care spending is known to be highly skewed, with a small subset of the population consuming a disproportionate amount of health care resources. Patients with cancer are high-cost users because of high incremental health care costs for treatment and the growing prevalence of cancer. The objectives of the present study included characterizing cancer-patient trajectories by cost, and identifying the patient and health system characteristics associated with high health system costs after cancer treatment. METHODS: This retrospective cohort study identified Ontario adults newly diagnosed with cancer between 1 April 2009 and 30 September 2010. Costs of health care use before, during, and after cancer episodes were used to develop trajectories of care. Descriptive analyses examined differences between the trajectories in terms of clinical and health system characteristics, and a logistic regression approach identified predictors of being a high-cost user after a cancer episode. RESULTS: Ten trajectories were developed based on whether patients were high- or low-cost users before and after their cancer episode. The most common trajectory represented patients who were low-cost in the year before cancer, survived treatment, and continued to be low-cost in the year after cancer (31.4%); stage ii cancer of the male genital system was the most common diagnosis within that trajectory. Regression analyses identified increases in age and in multimorbidity and low continuity of care as the strongest predictors of high-cost status after cancer. CONCLUSIONS: Findings highlight an opportunity to proactively identify patients who might transition to high-cost status after cancer treatment and to remediate that transition.

The case in favour of probiotics before, during and after pregnancy: insights from the first 1,500 days.

Successful human reproduction requires microbial homeostasis in the female reproductive tract, and colonisation of the newborn with beneficial microbes. In order to prevent several complications associated with dysbiosis, the administration of probiotics is more often being considered. The objective of the enclosed review was to examine the rationale for probiotic utility before and during pregnancy and in the early phase of infant life. The conclusions emerged from a panel of researchers who met during the International Scientific Association for Probiotics and Prebiotics (ISAPP) workshop held in Washington, DC, USA in 2015. The group concluded based upon the current literature, that a case can be made for the use of a specific sets of probiotic organisms during the first 1,500 days of life, with the goal of a healthy pregnancy to term, and a healthy start to life with lowered risk of infections and inflammatory events. The key to successfully translating these recommendations to practice is that products be made available and affordable to women in developed and developing countries.

Priapism of the Newborn - A Case Report.

Neonatal Priapism is a rare condition and its causes differ from that in childhood and adult. Management also is challenging as the ultimate goal is the preservation of normal erectile function. Most of the cases are idiopathic. A spontaneous detumescence occurs in majority of cases, so initially conservative non-surgical treatment is advocated. Here we report a case of a newborn presenting with priapism on the 1st day of life. Detumescence was achieved on the 8th day of life with conservative treatment.

Application of SCAR markers linked with mungbean yellow mosaic virus disease-resistance gene in Pakistan mungbean germplasm.

Mungbean yellow mosaic virus disease (MYMVD) caused by single-stranded DNA begomovirus is the most prominent threat to the mungbean crop in Pakistan. Mungbean genotypes consisting of 127 varieties/lines were screened for MYMVD under natural field conditions. No genotypes were found to be immune or highly resistant against MYMVD. Genotypes showing resistant and tolerant responses in field screening trials were screened using sequence characterized amplified region (SCAR) markers linked with the MYMVD-resistance gene. SCAR markers amplified the desired band only in the resistant and tolerant genotypes, while no amplification was observed in susceptible genotypes. SCAR markers will be useful for future breeding and varietal developmental programs and mungbean genotypes can be screened on the molecular level.

Supratentorial glioblastoma multiforme metastasizing to the cervical spinal cord.

Cases of glioblastoma multiforme (GBM) metastasizing to the leptomeninges or the intramedullary spine are rare and their prognosis are relatively poor. We present a case of supratentorial glioblastoma WHO grade IV which was later diagnosed to have cervical intramedullary metastasis 7 months after the primary surgery.

A young man with solitary trichoepithelioma of upper lip.

A 25-year-old man presented with a long-standing history of asymptomatic, skin-colored papule on cutaneous part of upper lip. Histopathologic examination of the papule demonstrated trichoepithelioma. The patient had no family history. He had also no history of similar lesion at same site or elsewhere. The patient was otherwise asymptomatic. Clinician excised the lesion as a part of treatment. Clinical differential diagnosis was sebaceous cyst and basal cell carcinoma.

Identification and authentication of Rosa species through development of species-specific SCAR marker(s).

Roses (Rosa indica) belong to one of the most crucial groups of plants in the floriculture industry. Rosa species have special fragrances of interest to the perfume and pharmaceutical industries. The genetic diversity of plants based on morphological characteristics is difficult to measure under natural conditions due to the influence of environmental factors, which is why a reliable fingerprinting method was developed to overcome this problem. The development of molecular markers will enable the identification of Rosa species. In the present study, randomly amplified polymorphic DNA (RAPD) analysis was done on four Rosa species, Rosa gruss-an-teplitz (Surkha), Rosa bourboniana, Rosa centifolia, and Rosa damascena. A polymorphic RAPD fragment of 391 bp was detected in R. bourboniana, which was cloned, purified, sequenced, and used to design a pair of species-specific sequence-characterized amplified region (SCAR) primers (forward and reverse). These SCAR primers were used to amplify the specific regions of the rose genome. These PCR amplifications with specific primers are less sensitive to reaction conditions, and due to their high reproducibility, these species-specific SCAR primers can be used for marker-assisted selection and identification of Rosa species.

Molecular characterization of urdbean (Vigna mungo) germplasm related to resistance against urdbean leaf crinkle virus.

Urdbean (Vigna mungo) is an important pulse crop grown worldwide. Urdbean leaf crinkle virus (ULCV) is a pathogen of urdbean found in Pakistan that causes huge losses in yield. Forty urdbean varieties/lines were screened against the virus under field conditions during spring season 2009. None of the lines appeared to be highly resistant or resistant. On the basis of a 0-5 disease rating scale and disease severity index, genotypes varied significantly in their reaction to ULCV. Four lines (M-6206, IAM-382-15, IAM-133, and Mash-1) were moderately resistant, eight were rated as moderately susceptible, and 21 as susceptible; the remaining seven lines were highly susceptible. RAPD analyses revealed an extensive amount of variation, which could be used for cultivar identification. Genetic differentiation among urdbean genotypes was similar to the field screening data. The varieties 6065-3 and 6206 were highly susceptible and moderately resistant, respectively, to ULCV under field conditions, confirmed by the RAPD analysis. These varieties were the most diverse varieties in the similarity matrix (67.2%), while the varieties IAM-382-9 and 07M003 were the most similar (98.4%). This information will help in the recognition of available resistant germplasms that can resist this disease and will be utilized for urdbean improvement in Pakistan.

Elucidation of thermotolerance diversity in cotton (Gossypium hirsutum L.) using physio-molecular approaches.

Cotton (Gossypium hirsutum) is an important cash crop, but high temperature during its growing season is one of the major factors that limit its productivity. This problem compels plant breeders to breed for heat tolerance, which can help to overcome this challenge. It is very important to make a comprehensive screening of heat-tolerant genotypes so that only the best are chosen. Here we report the combined use of several techniques that can help breeders to screen their germplasm. Twelve cultivated cotton genotypes were evaluated for thermotolerance, using assays that included electrolyte leakage, chlorophyll accumulation and protein profiling, as well as RAPDs to assess genetic diversity. Two genotypes (B-557 and NIAB-78) showed tolerant behavior in three thermotolerance assays. RAPD analysis results showed maximum similarity in a range of 86.7-66.7% between the genotypes MNH-554 and CIM-443. We conclude that combined use should be made of relative electrolyte leakage, chlorophyll stability and differential display with SDS-PAGE to aid in screening for stress tolerance. RAPD-based diversity analysis will further help to improve the efficiency of breeding programs.

Estimation of genetic distance based on RAPDs between 11 cotton accessions varying in heat tolerance.

The genetic distance of 11 cotton genotypes varying in heat tolerance was studied using RAPD markers. Fifty-three random decamer primers were used for the estimation of genetic distance. Among the 53 RAPD primers, which were custom synthesized by GeneLink Inc., UK, 32 were polymorphic and 21 were monomorphic. The 32 polymorphic primers produced 273 fragments, with a mean of 8.3 fragments per primer. The number of polymorphic bands produced in the 11 cotton accessions ranged from 1 to 31. Primer GLC-20 produced 31 polymorphic bands, while two primers, GLB-5 and GLC-12, produced one polymorphic band each. A range of 88.89 to 42.48% genetic similarity was observed among the 11 cotton accessions. The highest genetic similarity was observed between FH-945 and BH-160 (88.89%), whereas the lowest value was found between NIAB-801/2 and FH-945 (42.48%). Unique amplification profiles were produced by most of the cultivars; the differences were sufficient to distinguish them from other genotypes. This confirms the efficacy of RAPD markers for the identification of plant genotypes. An accumulative analysis of amplified products generated by RAPDs was sufficient to assess the genetic diversity among the genotypes. This information should be helpful for formulating breeding and genome mapping programs.

Evaluation of irradiation in foods using DNA Comet assay.

Comet assay is a rapid, inexpensive and sensitive biological technique to detect DNA damage in food stuffs by irradiation. In this study the Comet assay is applied on foods of plant and animal origins. Samples were irradiated by using Co-60 gamma-radiation source. The applied doses were 2, 6 and 10 kGy for food of plant origin and 0.5, 1 and 2 kGy for meat items. The un-irradiated and irradiated samples were clearly differentiated on the basis of DNA fragmentation. During the electrophoresis study, it was found that in un-irradiated cells DNA remained intact and appeared as Comets without tail whereas in irradiated cells Comets with tails were visible due to stretching of fragmented DNA. Moreover, it was also revealed that the DNA tail length was dose dependent. Dry food stuffs (seeds) showed good results as compared to moist foods (meat, fruits and vegetables) due to the absence of background damage.

Adults with chronic cough in urban Bangladesh: healthcare utilization and management of cases by private practitioners.

Tuberculosis (TB) case detection under the Directly Observed Treatment - Short-course (DOTS) strategy largely relies upon care seeking of chronic coughers and the actions taken by their healthcare providers. This study aimed to describe the healthcare utilization of people 15 years of age with a chronic cough in urban areas of Bangladesh and to understand their management by private practitioners. A community-based, household survey included 60,382 persons ≥ 15 years of age from two administrative areas of Dhaka City. A total of 1138 (2%) were identified to have had a cough for 3 weeks or more. This survey was linked to interviews of licensed and unlicensed practitioners in Dhaka and the Chittagong City Corporation of Bangladesh. Among identified coughers, 1046 (92%) were interviewed, of whom 648 (62%) reported having sought care from any provider. Among care seekers, 16% directly attended a DOTS facility. The remaining 84% sought care from the private sector, where less than 1% reported referral to a DOTS facility. Bivariate and multivariate assessments showed that care seeking from a licensed private practitioner or a DOTS centre was significantly associated with severity of the disease and previous diagnosis of TB.

Kala azar in Pregnancy.

Kala azar (KA) is one of the most important tropical diseases. More so is the issue of KA in pregnancy. In this retrospective study we tried to find out the out come of pregnant patient treated with Sodium Antimony Gluconate (SAG) for KA in one of the most endemic area of Bangladesh, Fulbaria. SAG is the available and registered drug for treatment of KA patient with pregnancy in Bangladesh. A total of 16 pregnant women presented with KA during the study period of 2005 to 2009. Out of the 16 patients 11 had abortion and they were all with in the 16 to 22nd week of pregnancy. The abortion took place mostly on the 22nd to 24th day of treatment when the patient had become afebrile. Rest of the 5 patients was in their 30 to 34th week of pregnancy and had good obstetric outcome. All the 16 patients were clinically cured at the end of treatment. No follow up records were available and there was no data regarding the 5 children. It is of our opinion that the abortions were induced by SAG and therefore we recommend that SAG should not be used in early or mid pregnancy for treating KA.