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Background: Structured training programs for robotic colorectal surgery are limited, and there are concerns about surgical outcomes and operating times., Objective: To compare perioperative and oncological outcomes of robotic total mesorectal excision for rectal cancer performed by expert consultants and surgical trainees in a modular surgical training program., Design: Retrospective cohort study., Settings: Conducted at a colorectal training referral center for robotic surgery., Patients: Consecutive robotic total mesorectal excision cases between May 2013 and December 2017 were evaluated retrospectively from a prospectively maintained institutional database and divided into 2 groups: group I comprised expert surgeons and group II comprised supervised trainees. Robotic total mesorectal excision training modules (5 modules) were performed stepwise with increasing complexity. Patients' demographic, perioperative, and oncological data were collected., Interventions: Modular robotic training., Main Outcome Measures: Comparable R0 resection rate, lymph node harvest, and oncological outcomes between experts and trainees, suggesting good quality in oncological resection., Results: A total of 177 robotic total mesorectal excision resections were performed (group I: n = 80, group II: n = 97). Four trainees completed 37.5 modules each. Patients' age, sex, and BMI were similar between groups. Group II had a higher ASA III score (6.3% vs 25.8%, p = 0.002). Clinical TNM and neoadjuvant chemoradiotherapy rates were similar. Group II had a longer operative time (225 [197.5-297.5] vs 250 [230-300] minutes, p = 0.004). No conversion occurred. There were no differences in intra- or postoperative outcomes between groups. The rate of R0 resection and the number of harvested lymph nodes were also similar between groups. The median follow-up was 75 (64.0-81.7) and 47 (38.0-55.0) months, respectively. Local and distant recurrence rates, 5-year overall survival (81.1% group I vs 81.3% group II, p = 0.832), and 5-year disease-free survival (79.7% group I vs 80.7% group II, p = 0.725) were similar between groups., Limitations: The groups operated in 2 consecutive periods., Conclusions: The robotic total mesorectal excision modular surgical training program maximizes training experience without significantly affecting the perioperative and oncological outcomes of patients with rectal cancer. See Video Abstract., El Impacto Del Programa Modular De Entrenamiento En Escisin Mesorrectal Total Robtica En Los Resultados Perioperatorios Y Oncolgicos En La Ciruga Robtica Del Cncer De Recto: ANTECEDENTES:Los programas de entrenamiento estructurados para la cirugía colorrectal robótica están limitados debido a preocupaciones sobre los resultados quirúrgicos y los tiempos de operación.OBJETIVO:Comparar los resultados perioperatorios y oncológicos de la escisión mesorrectal total robótica para el cáncer de recto realizada por consultores expertos y aprendices de cirugía en un programa modular de entrenamiento quirúrgica.DISEÑO:Estudio de cohorte retrospectivo.AJUSTES:Realizado en un centro de referencia de entrenamiento colorrectal para cirugía robótica.PACIENTES:Se evaluaron retrospectivamente casos consecutivos de escisión mesorrectal total robótica entre mayo de 2013 y diciembre de 2017 a partir de una base de datos institucional mantenida prospectivamente y se dividieron en dos grupos: Grupo I: cirujanos expertos; Grupo II: aprendices supervisados. Los módulos de entrenamiento robótico de escisión mesorrectal total (cinco módulos) se realizaron paso a paso con complejidad creciente. Se recogieron datos demográficos, perioperatorios y oncológicos.INTERVENCIONES:Entrenamiento modular en robótica.PRINCIPALES MEDIDAS DE RESULTADO:Tasa de resección R0 comparable, extracción de ganglios linfáticos y resultados oncológicos entre expertos y aprendices que sugieren buena calidad en la resección oncológica.RESULTADOS:Se realizaron un total de 177 resecciones por escisión mesorrectal total robótica (Grupo I: n = 80, Grupo II: n = 97). Cuatro alumnos completaron 37,5 módulos cada uno. La edad, el sexo y el IMC fueron similares entre los grupos. El grupo II tuvo una puntuación más alta de la Sociedad Americana de Anestesiólogos III (6,3% frente a 25,8%, p = 0,002). Las tasas clínicas de TNM y quimiorradioterapia neoadyuvante fueron similares. El grupo II tuvo mayor tiempo operatorio (225 (197,5-297,5) vs 250 (230-300) minutos, p = 0,004). No se produjo ninguna conversión. No hubo diferencias en los resultados intra o posoperatorios entre los grupos. La tasa de resección R0 y el número de ganglios linfáticos extraídos también fueron similares entre los grupos. La mediana de seguimiento fue de 75 (64,0-81,7) y 47 (38,0-55,0) meses, respectivamente. Tasas de recurrencia local y a distancia, supervivencia general a 5 años (81,1% Grupo I vs. 81,3% Grupo II, p = 0,832) y supervivencia libre de enfermedad a 5 años (79,7% Grupo I vs. 80,7% Grupo II, p = 0,725) fueron similares entre los grupos.LIMITACIONES:Los grupos operaron en dos períodos consecutivos.CONCLUSIONES:El programa de entrenamiento quirúrgico modular para la escisión mesorrectal total robótica maximiza la experiencia de capacitación sin afectar significativamente los resultados perioperatorios y oncológicos de los pacientes con cáncer de recto. (Traducción-Dr. Aurian Garcia Gonzalez)., (Copyright © The ASCRS 2024.)

The genus Phytophthora contains more than 100 plant pathogenic species that parasitize a wide range of plants, including economically important fruits, vegetables, cereals, and forest trees, causing significant losses. Global agriculture is seriously threatened by fungicide resistance in Phytophthora species, which makes it imperative to fully comprehend the mechanisms, frequency, and non-chemical management techniques related to resistance mutations. The mechanisms behind fungicide resistance, such as target-site mutations, efflux pump overexpression, overexpression of target genes and metabolic detoxification routes for fungicides routinely used against Phytophthora species, are thoroughly examined in this review. Additionally, it assesses the frequency of resistance mutations in various Phytophthora species and geographical areas, emphasizing the rise of strains that are resistant to multiple drugs. The effectiveness of non-chemical management techniques, including biological control, host resistance, integrated pest management plans, and cultural practices, in reducing fungicide resistance is also thoroughly evaluated. The study provides important insights for future research and the development of sustainable disease management strategies to counter fungicide resistance in Phytophthora species by synthesizing current information and identifying knowledge gaps., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

The current study was carried out to screen 10 isolates (ARS-01-ARS-10) of Rhizoctonia. solani from potato tubers cv. Kuroda, which were collected from various potato fields in Multan, Pakistan. The isolates were found to be morphologically identical, as the hyphae exhibit the production of branches at right angles and acute angles often accompanied by septum near the emerging branches. Anastomosis grouping showed that these isolates belonged to AG-3. A pathogenicity test was performed against the susceptible Kuroda variety and among the isolates, ARS-05 exhibited the highest mean severity score of approximately 5.43, followed by ARS-09, which showed a mean severity score of about 3.67, indicating a moderate level of severity. On the lower end of the severity scale, isolates ARS-06 and ARS-07 displayed mean severity scores of approximately 0.53 and 0.57, respectively, suggesting minimal symptom severity. These mean severity scores offer insights into the varying degrees of symptom expression among the different isolates of R. solani under examination. PCoA indicates that the severe isolate causing black scurf on the Kuroda variety was AG-3. A comprehensive analysis of the distribution, genetic variability, and phylogenetic relationships of R. solani anastomosis groups (AGs) related to potato crops across diverse geographic regions was also performed to examine AG prevalence in various countries. AG-3 was identified as the most widespread group, prevalent in Sweden, China, and the USA. AG-5 showed prominence in Sweden and the USA, while AG-2-1 exhibited prevalence in China and Japan. The phylogenetic analysis unveiled two different clades: Clade I comprising AG-3 and Clade II encompassing AG-2, AG-4, and AG-5, further subdivided into three subclades. Although AGs clustered together regardless of origin, their genetic diversity revealed complex evolutionary patterns. The findings pave the way for region-specific disease management strategies to combat R. solani's impact on potato crops.

Objective: To evaluate the subjects of possible ocular surface dysfunction in dry eye syndrome (DES) by using Ocular Surface Disease Index (OSDI) questionnaire and correlating it with the tear film break-up time (TBUT) test and Schirmer test., Study Design: Cross-sectional, observational study. Place and Duration of the Study: Armed Forces Institute of Ophthalmology (AFIO), National University of Medical Sciences (NUMS), Rawalpindi, Pakistan, from March to August 2022., Methodology:  Demographics and detailed ophthalmological examinations were carried out for all the patients using slit lamp biomicroscopy. The questionnaire for OSDI was filled to calculate the OSDI score, Schirmer test, and TBUT test were performed for all patients. For statistical analysis, the mean test score of both eyes was used. Correlations between tests were drawn and reported., Results: This study was conducted on ninety-seven adult participants with mean age of 31.3 ± 10.7 years, comprising of forty-five (46.4%) females and fifty-two (53.6%) males. The mean score for OSDI, TBUT, and Schirmer test was found to be 16.03 ± 14.22 (range 0 - 62.5), 9.63 ± 4.54 seconds (range 2.5 - 22.5), and 24.6 ± 10.85 mm (range 4.5 - 35.5), respectively. An inverse correlation was found between the OSDI and Schirmer, and OSDI and TBUT test scores which was also statistically significant. Schirmer and TBUT test scores also showed significant correlation., Conclusion: The OSDI is quick, precise, feasible for self-assessment, and non-invasive standardised tool for evaluating symptoms of dry eye disease, hence it can aid in the diagnosis of DES., Key Words: Dry eye syndrome, Ocular surface, Tear flim break-up time, Schirmer test.

Background: Babesia infections in sheep can cause a wide range of clinical and laboratory presentations. Changes in blood parameters are a meaningful manifestation of physiological and pathological changes in an organism., Aim: Therefore, the present study was conducted to analyze and compare hematological and biochemical parameters between blood profiles of Lohi sheep naturally infected and uninfected with Babesia ovis , the main causative agent of ovine babesiosis., Methods: Initially, blood and serum samples from 67 Lohi sheep were collected, DNA was extracted and babesial infection was detected through polymerase chain reaction. The overall infection rate of B. ovis was 37% (25/67). Sixteen infected (experiment group) and 16 uninfected (control group) sheep that were apparently healthy with no history of previous treatment for babesiosis, were selected for hemato-biochemical analysis. Blood samples were analyzed through an automatic CBC analyzer, while serum collected from gel vacutainers was analyzed for blood urea, blood urea nitrogen (BUN), creatinine, and total bilirubin. Each parameter was compared between infected and uninfected animals using a paired t -test in Minitab Express™ software for statistical analyses., Results: Erythron comparison showed a highly significant ( p < 0.0001) decrease in RBC, hemoglobin, and Hct. A nonsignificant increase in mean corpuscular volume (MCV), red cell distribution width (RDW), and RDW-standard deviation (RDW-SD), while a nonsignificant decrease in mean corpuscular haemoglobin (MCH) and MCH concentration (MCHC) values was recorded in infected sheep. Leukon comparison showed a significantly low level of total leukocyte ( p < 0.001) in infected sheep. Platelet (Plt) along with platelet crit (Pct) and platelet distribution width (PDW) were nonsignificantly higher, whereas a nonsignificant decrease in mean Plt volume was recorded in infected sheep as compared to uninfected animals. Among biochemical parameters, blood urea, BUN, and total bilirubin showed significant differences ( p < 0.05), while creatinine showed a nonsignificant difference., Conclusion: To the best of our knowledge, this is the first report on hemato-biochemical changes associated with babesiosis in the Lohi breed. Consistent with hemolytic anemia, these data would justify physical examination and, together with the medical history, would provide an excellent basis for the diagnosis of babesiosis., Competing Interests: The authors declare no conflict of interest.

Heavy metals contamination is posing severe threat to the soil health and environmental sustainability. Application of industrial and sewage waste as irrigation and growing urbanization and agricultural industry is the main reason for heavy metals pollution. Therefore, the present study was planned to assess the influence of different irrigation sources such as industrial effluents, sewage wastewater, tube well water, and canal water on the soil physio-chemical, soil biological, and enzymatic characteristics. Results showed that sewage waste and industrial effluents affect the soil pH, organic matter, total organic carbon, and cation exchange capacity. The highest total nickel (383.71 mg kg -1 ), lead (312.46 mg kg -1 ), cadmium (147.75 mg kg -1 ), and chromium (163.64 mg kg -1 ) were recorded with industrial effluents application. Whereas, industrial effluent greatly reduced the soil microbial biomass carbon (SMB-C), soil microbial biomass nitrogen (SMB-N), soil microbial biomass phosphorus (SMB-P), and soil microbial biomass sulphur (SMB-S) in the winter season at sowing time. Industrial effluent and sewage waste inhibited the soil enzymes activities. For instance, the minimum activity of amidase, urease, alkaline-phosphatase, β-glucosidase, arylsulphatase and dehydrogenase activity was noted with HMs contamination. The higher levels of metals accumulation was observed in vegetables grown in soil contaminated with untreated waste water and industrial effluent in comparison to soil irrigated with canal and tube well water. The mean increase in soil microbial parameters and enzyme activities was also observed in response to the change in season from winter to spring due to increase in soil mean temperature. The SMB-C, SMB-N, SMB-P and SMB-S showed significant positive correlation with soil enzymes (amidase, urease, alkaline-phosphatase, β-glucosidase, arylsulphatase and dehydrogenase). The heavy metals accumulation in soil is toxic to microorganisms and inhibits enzyme functions critical for nutrient cycling and organic matter decomposition and can disrupt the delicate balance of soil ecosystem and may lead to long-term damage of soil biological health., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Objective: To compare and correlate corneal keratometry (K), central corneal thickness (CCT), and anterior chamber depth (ACD) using a partial coherence interferometry device and a Scheimpflug camera system device in patients planned for cataract surgery., Study Design: Comparative cross-sectional study. Place and Duration of the Study: Armed Forces Institute of Ophthalmology, from December 2021 to June 2022., Methodology: Patients planned for cataract surgery underwent measurement of central corneal thickness (CCT), anterior chamber depth (ACD), and keratometric values (K) by Wavelight OB 820 biometer and Allergo Oculyzer II, preoperatively. Correlation and inter-device agreement were assessed between two devices., Results: There were 115 patients with a mean age of 64.23 ± 9.26 years. All anterior segment parameters showed excellent agreement and high degree of correlation between the two optical devices. Correlation coefficient (r) for central corneal thickness was r = 0.958 (p<0.001) while correlation coefficient for anterior chamber depth was r = 0.965 (p<0.001), r = 0.966 (p<0.001) for flat K, and r = .0969 (p<0.001) for steep keratometry, respectively., Conclusion: Ophthalmic parameters assessed by both devices were comparable and are recommended to be used in the clinical practice of keratorefractive and cataract surgeries interchangeably., Key Words: ACD, CCT, Keratometry, Cataract, OB 820 biometer, Oculyzer II.

Rhizoctonia solani is one of the most common soil-borne fungal pathogens of legume crops worldwide. We collected rDNA-ITS sequences from NCBI GenBank, and the aim of this study was to examine the genetic diversity and phylogenetic relationships of various R. solani anastomosis groups (AGs) that are commonly associated with grain legumes (such as soybean, common bean, pea, peanut, cowpea, and chickpea) and forage legumes (including alfalfa and clover). Soybean is recognized as a host for multiple AGs, with AG-1 and AG-2 being extensively investigated. This is evidenced by the higher representation of sequences associated with these AGs in the NCBI GenBank. Other AGs documented in soybean include AG-4, AG-7, AG-11, AG-5, AG-6, and AG-9. Moreover, AG-4 has been extensively studied concerning its occurrence in chickpea, pea, peanut, and alfalfa. Research on the common bean has been primarily focused on AG-2, AG-4, and AG-1. Similarly, AG-1 has been the subject of extensive investigation in clover and cowpea. Collectively, AG-1, AG-2, and AG-4 have consistently been identified and studied across these diverse legume crops. The phylogenetic analysis of R. solani isolates across different legumes indicates that the distinct clades or subclades formed by the isolates correspond to their specific anastomosis groups (AGs) and subgroups, rather than being determined by their host legume crop. Additionally, there is a high degree of sequence similarity among isolates within the same clade or subclade. Principal coordinate analysis (PCoA) further supports this finding, as isolates belonging to the same AGs and/or subgroups cluster together, irrespective of their host legume. Therefore, the observed clustering of R. solani AGs and subgroups without a direct association with the host legume crop provides additional support for the concept of AGs in understanding the genetic relationships and evolution of R. solani .

Agrochemicals are products of advanced technologies that use inorganic pesticides and fertilizers. Widespread use of these compounds has adverse environmental effects, leading to acute and chronic exposure. Globally, scientists are adopting numerous green technologies to ensure a healthy and safe food supply and a livelihood for everyone. Nanotechnologies significantly impact all aspects of human activity, including agriculture, even if synthesizing certain nanomaterials is not environmentally friendly. Numerous nanomaterials may therefore make it easier to create natural insecticides, which are more effective and environmentally friendly. Nanoformulations can improve efficacy, reduce effective doses, and extend shelf life, while controlled-release products can improve the delivery of pesticides. Nanotechnology platforms enhance the bioavailability of conventional pesticides by changing kinetics, mechanisms, and pathways. This allows them to bypass biological and other undesirable resistance mechanisms, increasing their efficacy. The development of nanomaterials is expected to lead to a new generation of pesticides that are more effective and safer for life, humans, and the environment. This article aims to express at how nanopesticides are being used in crop protection now and in the future. This review aims to shed some light on the various impacts of agrochemicals, their benefits, and the function of nanopesticide formulations in agriculture., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Masson SAS. All rights reserved.)

Elicitors are stressors that activate secondary pathways that lead to the increased production of bioactive molecules in plants. Different elicitors including the fungus Aspergillus niger (0.2 g/L), methyl jasmonate (MeJA, 100 µM/L), and silver nanoparticles (1 µg/L) were added, individually and in combination, in a hydroponic medium. The application of these elicitors in hydroponic culture significantly increased the concentration of photosynthetic pigments and total phenolic contents. The treatment with MeJA (methyl jasmonate) (100 µM/L) and the co-treatment of MeJA and AgNPs (silver nanoparticles) (100 µM/L + 1 µg/L) exhibited the highest chlorophyll a (29 µg g -1 FW) and chlorophyll b (33.6 µg g -1 FW) contents, respectively. The elicitor MeJA (100 µM/L) gave a substantial rise in chlorophyll a and b and total chlorophyll contents. Likewise, a significant rise in carotenoid contents (9 µg/g FW) was also observed when subjected to meJA (100 µM/L). For the phenolic content, the treatment with meJA (100 µM/L) proved to be very effective. Nevertheless, the highest production (431 µg/g FW) was observed when treated with AgNPs (1 µg/L). The treatments with various elicitors in this study had a significant effect on flavonoid and lignin content. The highest concentration of flavonoids and lignin was observed when MeJA (100 mM) was used as an elicitor, following a 72-h treatment period. Hence, for different plant metabolites, the treatment with meJA (100 µM/L) and a co-treatment of MeJA and AgNPs (100 µM/L + 1 µg/L) under prolonged exposure times of 120-144 h proved to be the most promising in the accretion of valuable bioactive molecules. The study opens new insights into the use of these elicitors, individually or in combination, by using different concentrations and compositions.

Citrus viroid infection is emerging as a serious threat because of its efficient systemic movement within the host plant and its quick spread due to contaminated pruning tools. A survey was conducted to investigate the primary distribution and molecular characterization of Citrus bent leaf viroid (CBLVd) and its variants in different citrus cultivars. A total of 154 symptomatic citrus samples were collected and detected by RT‒PCR with newly designed specific primers with the incidence of 36.33%. During biological indexing study on Etrog citron, expressions of reduced leaf size, yellowing with a light green pattern, and bending were observed. Amplified products were sequenced and analyzed using a nucleotide BLAST search, which showed 98% homology with other CBLVd isolates. The results of the phylogenetic tree analysis showed the presence of two main groups (A and B), with the predominant variants of CBLVd, i.e., CVd-I-LSS (Citrus viroid Low Sequence Similarity) sequences, clustering in subgroup A1 along with newly detected CVd-I-LSS from Palestinian sweet lime (Citrus limettioides), which has been identified as a new host of CVd-I-LSS in Pakistan. Further analysis of the sequences in subgroup A1 showed that the variant of CVd-I-LSS infecting citrus cultivars had a close relationship with isolates reported from China, Japan, and Iran, which may have resulted from the exchange of planting material. This study also unveiled the variability in nucleotide sequences of CBLVd, which made it unable to be detected by old primers. The results of this study indicate that the widespread presence of divergent variants of CBLVd is a major concern for the citrus industry in Pakistan and other countries where virulent isolates of CBLVd are prevalent. These findings suggest the need for future research on effective management and quarantine measures to stop the spread of CBLVd., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Ali, Umar, Naqvi, Shakeel, Tahir, Khan, Altaf, Ölmez, Dababat, Haq, Nadeem, Hatipoğlu, Baloch and Chung.)

Rhizoctonia solani is a species complex composed of many genetically diverse anastomosis groups (AG) and their subgroups. It causes economically important diseases of soybean worldwide. However, the global genetic diversity and distribution of R. solani AG associated with soybean are unknown to date. In this study, the global genetic diversity and distribution of AG associated with soybean were investigated based on rDNA-ITS sequences deposited in GenBank and published literature. The most prevalent AG, was AG-1 (40%), followed by AG-2 (19.13%), AG-4 (11.30%), AG-7 (10.43%), AG-11 (8.70%), AG-3 (5.22%) and AG-5 (3.48%). Most of the AG were reported from the USA and Brazil. Sequence analysis of internal transcribed spacers of ribosomal DNA separated AG associated with soybean into two distinct clades. Clade I corresponded to distinct subclades containing AG-2, AG-3, AG-5, AG-7 and AG-11. Clade II corresponded to subclades of AG-1 subgroups. Furthermore, AG and/or AG subgroups were in close proximity without corresponding to their geographical origin. Moreover, AG or AG subgroups within clade or subclades shared higher percentages of sequence similarities. The principal coordinate analysis also supported the phylogenetic and genetic diversity analyses. In conclusion, AG-1, AG-2, and AG-4 were the most prevalent AG in soybean. The clade or subclades corresponded to AG or AG subgroups and did not correspond to the AG's geographical origin. The information on global genetic diversity and distribution will be helpful if novel management measures are to be developed against soybean diseases caused by R. solani ., Competing Interests: The authors declare no conflict of interest.

Background Peripheral neuropathy is a controversial but serious complication of beta-thalassemia (β-Th). Although few studies have reported no relationship between neuropathy and thalassemia, many have linked it with increasing age, iron overload, and iron chelator toxicity. This study aims to investigate the presence of neuropathy in β-Th using corneal nerve fibers. Methodology A cross-sectional study was conducted using corneal confocal microscopy on individuals with intermediate and major β-Th who were compared to healthy individuals. The main outcome variables were corneal main nerve and branch nerve densities which were calculated using Image J software. The comparison between groups was done using the independent-samples F-test and Bonferroni post-hoc analysis. Results There was reduced corneal main nerve and branch nerve density in β-Th intermediate and major patients compared to the control group, and the results were statistically significant (p-value <0.05). However, a significant correlation was not observed between serum ferritin levels and corneal nerve parameters. Conclusions The reduction in corneal nerve parameters in β-Th patients compared to healthy controls can be an indication of peripheral neuropathy in β-Th. Further work is needed to confirm these findings., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2022, Khan et al.)

Nanotechnology is a burning field of scientific interest for researchers in current era. Diverse plant materials are considered as potential tool in green chemistry based technologies for the synthesis of metal nanoparticles (NPs) to cope with the hazardous effects of synthetic chemicals, leading to severe abiotic climate change issues in today's agriculture. This study aimed to determine the synthesis and characterization of metal-based nanoparticles using extracts of the selected plant Calotropis gigantea and to evaluate the enzyme-inhibition activities and antibacterial and antifungal activity of extracts of metal-based zinc nanoparticles using C. gigantea extracts. The crystal structure and surface morphology were characterized by X-ray diffraction (XRD) and scanning electron microscopy (SEM). C. gigantea was examined for antimicrobial activity against clinical isolates of bacteria and fungi. The water, ethanolic, and acetone extracts of C. gigantea were studied for their antagonistic action against bacterial strains ( E. coli , S. aureus , P. multocida , and B. subtilis ) and selected fungal strains ( A. paracistic , F. solani , A. niger , S. ferrugenium, and R. nigricans ). In vitro antimicrobial activity was determined by the disc diffusion method, where C. gigantea wastested for AChE and BChE inhibitory activity using Ellman's methodology. The kinetic analysis was performed by the proverbial Berthelot reaction for urease inhibition. The results showed that out of all the extracts tested, ethanolic and water extracts possessed zinc nanoparticles. These extracts showed the maximum zone of inhibition against F. solani and P. multocida and the lowest against S. ferrugenium and B. subtilis . A potential source of AChE inhibitors is certainly provided by the abundance of plants in nature. Numerous phyto-constituents, such as AChE and BChE inhibitors, have been reported in this communication. Water extract was active and has the potential for in vitro AChE and BChE inhibitory activity. The urease inhibition with flower extracts of C. gigantea revealed zinc nanoparticles in water extracts that competitively inhibited urease enzymes. In the case of cholinesterase enzymes, it was inferred that the water extract and zinc nanoparticles have more potential for inhibition of BChE than AChE and urease inhibition. Furthermore, zinc nanoparticles with water extract are active inthe inhibition of the bacterial strains E. coli , S. aureus , and P. multocida and the fungal strains A. paracistic , F. solani , and A. niger .

Bitter gourd (Momordica charantia L.) is an important vegetable crop of the Cucurbitaceae family widely cultivated in Pakistan and around the world. In October 2020, a nutrition management trial of Bitter gourd cv. Seminis-200) was conducted on an area of 10,860 sq. ft. (99×110 feet) at the Agricultural Research farm of Bahauddin Zakariya University, Multan (30.2601° N, 71.5158° E), Pakistan. Symptoms of large, brown necrotic leaf spots were observed on the leaves of bitter gourd vines. The disease started from the yellowing of leaves within the reticulate venation and turned brown. Irregular brown leaf spots coalesced to form large necrotic areas followed by foliar chlorosis then wilting that occurred very late. There were no crown rot symptoms although there was slight discoloration of roots and when cut longitudinally, browning of tissues was observed. The disease was assessed visually with 37% incidence which resulted in poor quality and yield in terms of reduced size and yellowing of fruit. Infected vines along with the roots were collected for the isolation of pathogen. A total of 34 leaves and 22 root samples were collected from the field for isolation. The leaf, collar and root portions were cut into 0.5 to 1 cm in length and surface disinfected with 1% sodium hypochlorite (NaOCl) for 2-3 minutes followed by washing twice with autoclaved distilled water and after drying, placed on potato dextrose agar (PDA) medium, and incubated at 25±2 °C for one week. The fungal colonies of fluffy white growth with light orange pigment were isolated. For morphological characterization, a total of 4 pure cultures were isolated from leaves, collar region and root by single spore technique on carnation leaf agar (CLA) medium after 15 days of incubation at 25±2℃. Curved and thick-walled macroconidia with elongated or pointed apical characteristic foot-shaped basal cells were produced in sporodochia. Macroconidia with 5-7 septa measured 22.50-41.80 μm × 2.90-4.20 μm (n = 60). Thick, brown with roughened walls and subglobose ellipsoidal chlamydospores were observed in clumps or chains with the dimension of 5.8 to 10.8 μm (n = 20). On morphological characteristics, the fungus was identified as Fusarium equiseti (Corda) Sacc. according to Leslie and Summerell (2006). Two single spore isolates were used for molecular identification by amplifying ribosomal DNA of the internal transcribed spacer (ITS) region with ITS1/ITS4 primers (White et al. 1990) and for β-tubulin gene region, primers T1/Bt-2b (O'Donnell and Cigelnik, 1997) were used. The obtained sequences were deposited in GenBank with accession numbers MW880179 and MW880198 from the ITS region and BLAST search in GenBank showed 100 and 98.11% alignment with previously published sequences of F. equiseti with accessions OM992323.1and MT558569.1 respectively. Accession number OM867571from the β-tubulin region showed 100% sequence similarity with F. equiseti with accession MN653163.1. For pathogenicity, macroconidia from 2-week-old cultures on CLA medium were harvested to prepare spore suspension (1 × 106 conidia/ml). Koch's postulates were confirmed on nine bitter gourd plants (cv. Seminis-200) by applying spore suspension of fungal inoculum at 3-4 leaf stage separately on leaves by automizer, on collar region after making incision spore suspension was applied and in the root zone, 20ml spore suspension was added whereas distilled water was used as a control with three replications. Plants were kept under controlled conditions in the greenhouse with 65% to 75% humidity and the temperature was maintained at 32±2 °C for one week. After 7-8 days, inoculated plants began to exhibit symptoms of brown, necrotic leaf spots on the leaves of bitter gourd vines followed by yellowing of leaves that eventually turned brown. Roots showed slight discoloration and browning of vascular bundles and finally, the plants wilted after four weeks. while control plants remained symptomless. The symptoms resembled those noticed in the field. The fungus was re-isolated from leaves, collar region and roots, followed by morphological identification, and finally confirmed as F. equiseti. To the best of our knowledge, this is the first report of a leaf spot caused by F. equiseti in a bitter gourd from Pakistan. If the disease is not managed properly, it may cause a drastic effect on yield under favorable environmental conditions. The pathogen may also damage other cucurbitaceous crops cultivated in the area.

Plant tissue culture technique employed for the identification and isolation of bioactive phytocompounds has numerous industrial applications. It provides potential benefits for different industries which include food, pharmaceutical and cosmetics. Various agronomic crops i.e., cereals, fruits, vegetables, ornamental plants and forest trees are currently being used for in vitro propagation. Plant tissue culture coupled with biotechnological approaches leads towards sustainable agricultural development providing solutions to major food security issues. Plants are the rich source of phytochemicals with medicinal properties rendering them useful for the industrial production of pharmaceuticals and nutraceuticals. Furthermore, there are numerous plant compounds with application in the cosmetics industry. In addition to having moisturizing, anti-ageing, anti-wrinkle effects; plant-derived compounds also possess pharmacological properties such as antiviral, antimicrobial, antifungal, anticancer, antioxidant, anti-inflammatory, and anti-allergy characteristics. The in vitro propagation of industrially significant flora is gaining attention because of its several advantages over conventional plant propagation methods. One of the major advantages of this technique is the quick availability of food throughout the year, irrespective of the growing season, thus opening new opportunities to the producers and farmers. The sterile or endangered flora can also be conserved by plant micro propagation methods. Hence, plant tissue culture is an extremely efficient and cost-effective technique for biosynthetic studies and bio-production, biotransformation, or bioconversion of plant-derived compounds. However, there are certain limitations of in-vitro plant regeneration system including difficulties with continuous operation, product removal, and aseptic conditions. For sustainable industrial applications of in-vitro regenerated plants on a large scale, these constraints need to be addressed in future studies., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Hasnain, Naqvi, Ayesha, Khalid, Ellahi, Iqbal, Hassan, Abbas, Adamski, Markowska, Baazeem, Mustafa, Moustafa, Hasan and Abdelhamid.)

Cotton ( Gossypium hirsutum ) is a global cash crop which has gained importance in earning foreign exchange for each country. Bacterial blight caused by Xanthomonascitri subsp. malvacearum ( Xcm ) has been a seriousdisease in Pakistan's cotton belt on multiple occasions. Bacterium was isolated and identified through various biochemical and diagnostic tests wherehypersensitivity reaction, Gram staining, KOH (potassium hydroxide), catalase, starch hydrolysis, lecithinase and Tween 80 hydrolysis tests confirmed bacterium as Gram-negative and plant pathogenic. Xcm perpetuation assays wereevaluated on various cotton varieties under glasshouse conditions in completely randomized design by three different methods, wherein the scratch method proved to be the best upon CIM-496 and showed 83.33% disease incidence as compared with the other two methods, where Bt-3701 responded with 53.33% incidence via the spray gun method, and 50% with the water splash method on CIM-616, as compared with the control. Similarly, for disease severity percentage, Bt-3701 was pragmatic with 47.21% through scratch method, whereas, in the spray gun method, 45.51% disease severity was noted upon Bt-802, and 31.27% was calculated on Cyto-179 through the water splash method. Owing to the unique antibacterial properties of aqueous plant extracts, the poison food technique showed Aloe vera , Mentha piperita , Syzygiumcumini and Azadirachta indica with 17.77, 29.33, 18.33 and 20.22 bacterial colonies counted on nutrient agarmedium petri plate, respectively, as compared with the control. Measurement of the inhibition zone by disk diffusion technique showed Mentha piperita , Syzygiumcumini , Citrus limon , Moringa oleifera and Syzygium aromaticum to present the most promising results by calculating the maximum diameter of the inhibition zone, viz., 8.58, 8.55, 8.52, 8.49 and 8.41 (mm), respectively, at the highest tested concentration (75 ppm, parts per million) compared with the control. It is probable that the decoction's interaction with the pathogen population on the host plant will need to be considered in future experiments. However, at this moment, more research into the effective management of cotton bacterial blight by plant extracts in terms of concentration determination and development of biopesticides will provide future avenues to avoid environmental pollution.

Plants often face incompatible growing environments like drought, salinity, cold, frost, and elevated temperatures that affect plant growth and development leading to low yield and, in worse circumstances, plant death. The arsenal of versatile compounds for plant consumption and structure is called metabolites, which allows them to develop strategies to stop enemies, fight pathogens, replace their competitors and go beyond environmental restraints. These elements are formed under particular abiotic stresses like flooding, heat, drought, cold, etc., and biotic stress such as a pathogenic attack, thus associated with survival strategy of plants. Stress responses of plants are vigorous and include multifaceted crosstalk between different levels of regulation, including regulation of metabolism and expression of genes for morphological and physiological adaptation. To date, many of these compounds and their biosynthetic pathways have been found in the plant kingdom. Metabolites like amino acids, phenolics, hormones, polyamines, compatible solutes, antioxidants, pathogen related proteins (PR proteins), etc. are crucial for growth, stress tolerance, and plant defense. This review focuses on promising metabolites involved in stress tolerance under severe conditions and events signaling the mediation of stress-induced metabolic changes are presented., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Chen, Mubeen, Hasnain, Rizwan, Adrees, Naqvi, Iqbal, Kamran, El-Sabrout, Elansary, Mahmoud, Alaklabi, Sathish and Din.)

Plants in nature are under the persistent intimidation of severe microbial diseases, threatening a sustainable food production system. Plant-bacterial pathogens are a major concern in the contemporary era, resulting in reduced plant growth and productivity. Plant antibiotics and chemical-based bactericides have been extensively used to evade plant bacterial diseases. To counteract this pressure, bacteria have evolved an array of resistance mechanisms, including innate and adaptive immune systems. The emergence of resistant bacteria and detrimental consequences of antimicrobial compounds on the environment and human health, accentuates the development of an alternative disease evacuation strategy. The phage cocktail therapy is a multidimensional approach effectively employed for the biocontrol of diverse resistant bacterial infections without affecting the fauna and flora. Phages engage a diverse set of counter defense strategies to undermine wide-ranging anti-phage defense mechanisms of bacterial pathogens. Microbial ecology, evolution, and dynamics of the interactions between phage and plant-bacterial pathogens lead to the engineering of robust phage cocktail therapeutics for the mitigation of devastating phytobacterial diseases. In this review, we highlight the concrete and fundamental determinants in the development and application of phage cocktails and their underlying mechanism, combating resistant plant-bacterial pathogens. Additionally, we provide recent advances in the use of phage cocktail therapy against phytobacteria for the biocontrol of devastating plant diseases.

The study objective was to assess the correlation of time outdoors in sunlight hours with spherical equivalent refraction (SER) and other variables. This was a cross-sectional study carried out in the Fauji Foundation Hospital, Rawalpindi, between January and April 2019. A total of 110 Individuals were included via convenient sampling, with myopia defined as having at least -0.5 D (SER ≤ -0.5 D). Collection of Questionnaires with interviews regarding eye care behaviour along with auto-refraction was done, followed by calculation of spherical equivalent refraction. Those having high myopia of 6 D or more (SER ≤ - 6 D), hyperopia of 0.5 D or more (SER ≥ +0.5 D), and history significant for ocular trauma and surgeries, were excluded. After data cleaning and their entry, analyses were done using SPSS version 26. The mean age data, available for 105 individuals, was 15.4 ± 3.69 years. Spearman's correlation was used for right SER with time outdoors in summers (r= +0.25, p=0.008) and winters (r= +0.243, p=0.010), indicating an inverse relationship with myopia. Females had more myopic refraction than males, but the independent sample t-test was not significant. Logistic regression was used and a protective effect was found for both summer and winter time outdoors, while education level was associated with myopia. Key Words: Myopia, Spherical equivalent refraction (SER), Sunlight exposure.

Background  Minimal access surgery is associated with improved cosmetic and other short-term outcomes. Conventionally, an abdominal incision is made for specimen extraction. We assessed the feasibility of specimen extraction through one of the natural orifices and analyzed its impact on short-term outcomes. Methods  A prospectively collected data were reviewed on consecutive patients who underwent natural orifice specimen extraction (NOSE) after laparoscopic colorectal surgery. The results were compared with a matched group who had transabdominal extraction (TAE) of the specimens. A systematic literature review was performed to compare our results. Results  The combined median operating time for right and left colectomies was significantly higher in the NOSE group as compared with TAE group (260 vs. 150). There was no mortality in either group and no conversions to TAE in the NOSE group. No local metastasis or major iatrogenic injuries were reported at the time of retrieval. The results were comparable to those of a meta-analysis of randomized controlled trials. Conclusion  The results of NOSE are comparable to those of TAEs. The absence of a minilaparotomy for specimen extraction may lead to a speedy recovery and better cosmesis., Competing Interests: Conflict of Interest None declared., (The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. ( https://creativecommons.org/licenses/by/4.0/ ).)

Objective: To evaluate the success rate of Transcanalicular Diode laser assisted Dacryocystorhinostomy (TC-DCR) in cases of acquired nasolacrimal duct obstruction (NLDO)., Methods: This Quasi-Experimental study was conducted in Armed Forces Institute of Ophthalmology (AFIO) Rawalpindi, from July 2018 to April 2019. In this study, 73 patients of chronic dacryocystitis secondary to acquired NLDO were treated by TC-DCR under general anaesthesia. Follow up examination was done after one day, one week and three months. Silicone tubes were removed three months after surgery and syringing of lacrimal system done to confirm patency of lacrimal passages. Success of the procedure was documented as absence of epiphora and patent nasolacrimal duct on syringing., Results: Seventy three patients (males 27; females 46) were included in this study. Mean age of these patients was 51.6+21 years. On completion of this study which was three months after surgery, we found subjective improvement (absence of epiphora) in 86.3% patients and objective improvement (successful irrigation of lacrimal passages) in 93.2% patients., Conclusion: TC- DCR is a minimally invasive technique of doing Dacryocystorhinostomy and imparts more than 90% success rate. It has additional advantages of good cosmetic results, low complication rate and short surgery and convalescence time., Competing Interests: Conflict of Interest: No conflict of Interest., (Copyright: © Pakistan Journal of Medical Sciences.)

Citrus tristeza virus (CTV) by killing millions of citrus cultivars grown on sour orange rootstock worldwide has become one of the most dangerous viral pathogen. Characterization of 12 CTV isolates was analyzed by biological indexing. Infected samples of citrus were collected from citrus growing areas of Pakistan and CTV was detected by symptoms on indicator plants and confirmed by direct tissue blot immunoassay (DTBIA). CTV positive samples were graft inoculated on six biological indicator hosts in the study. A standardized protocol was deployed to study biological characteristics of these isolates. All biological indicators induced mild and from mild to moderate reactions against all of the CTV isolates tested. About two isolates produced stem-pitting symptoms from moderate to severe on Mexican lime. CTV strains were further characterized and confirmed by the analysis of p25 gene of CTV isolates using single-strand conformation polymorphism (SSCP) assay. SSCP analysis revealed that most isolates confined only one predominant sequence variant. SSCP profiles of PCR amplified products from CTV isolates showed bands patterns corresponding to mild and sever strain. Three isolates (4MF, 8KBS and 10GS) from different regions and cultivars were identified as potential source of mild strains for cross protection. These results are the best base for mild strain cross protection (MSCP) in the country.

Huanglongbing (HLB), also known as citrus greening disease, is the most devastating disease of citrus across the world, caused by the phloem limited fastidious bacterium 'Candidatus Liberibacter spp.'. This research was conducted on HLB infected 10-year-old Kinnow orchard located at Multan, Pakistan. Different classes of antibiotics in various combinations were applied on HLB-infected trees. The antibiotic treatments were applied before flowering in February, during fruit setting in April and at fruit growth stage in June. The different antibiotics combinations used were Ampicillin sodium + Rifampicin, Cefalexin + Rifampicin, Ampicillin sodium + Cefalexin, Ampicillin sodium + Cefalexin + Rifampicin and Control (distilled water). Different fruit qualitative and quantitative attributes were examined. The application of antibiotics significantly decreased 2-11% in flower, June and pre-harvest drops as compared to control. Further, antibiotics increased fruit weight and yield by five times while the juice content, total soluble solids, ripening index, total sugars, phenolic and vitamin C content were also increased in fruits. In addition, total soluble proteins, peroxidase and catalase activities were increased in fruits harvested from antibiotic treated plants compared to control, however the superoxidase dismutase activity was decreased in fruits of antibiotic treated plants. Finally, it is concluded that application of different antibiotics combinations helps in improving the fruit yield and different quality attributes of HLB infected Kinnow trees.

Objective: To determine the role of hypertension, hyperlipidemia, smoking and positive family history of diabetes and hypertension in the development of diabetic retinopathy., Methods: This prospective cohort study was conducted at the Department of Chemical Pathology, Armed Forces Institute of Pathology, Rawalpindi over 2 years period from June 2014 to June 2016. One hundred consecutive diabetic patients with no signs of diabetic retinopathy and good glycemic control (HbA1c<6.5%) were registered by non-probability convenient sampling after taking written informed consent. They were evaluated for hypertension, hyperlipidemia and smoking status. These patients were then followed 6 monthly for 2 years to look for the development of diabetic retinopathy., Results: The mean age of the patients was 50.72±9.29 years and there were 57 (57%) male and 43 (43%) female patients. Majority (82%) of the patients had NIDDM. The mean duration of diabetes was 8.31±6.83 years. 11% of the patients were smoker, 37% were hypertensive, 6% had hyperlipidaemia, 62% had family history of diabetes and 30% had family history of hypertension. At the end of follow-up, 9 (9.0%) patients had diabetic retinopathy. The frequency of diabetic retinopathy increased with increasing age of the patient; however, the difference was statistically insignificant. A comparatively higher frequency of diabetic retinopathy was also seen in patients with IDDM and those with positive family history of diabetes and hypertension yet again, the difference was statistically insignificant. Also, no significant difference was noted among male and female genders and smokers vs. non-smoker. However, the frequency of diabetic retinopathy increased significantly with increasing duration of diabetes. It was also higher among those with hypertension and hyperlipidemia., Conclusion: Higher patient age (≥50 years), increasing duration of diabetes (≥20 years), insulin dependent diabetes mellitus, hypertension, hyperlipidemia, and positive family history of diabetes and hypertension were found to be associated with increased frequency of diabetic retinopathy.

Aims  Rectal prolapse is a debilitating and unpleasant condition adversely affecting the quality of life. Laparoscopic ventral mesh rectopexy (LVMR) is recognized as one of the treatment options. The aim of this study was to evaluate the functional outcomes after a standardized LVMR. Methods  A cohort of patients who underwent LVMR from 2011 to 2015 were contacted and asked to fill questionnaires about their symptoms before and after the surgery. Three questionnaires based on measurement of Wexner fecal incontinence (WFI), obstructive defecation syndrome (ODS), and Birmingham Bowel and Urinary Symptom (BBUS) scores were used to assess the changes in postoperative functional outcomes. Some additional questions were also added to further assess bowel dysfunction. Results  There were 58 female patients with a mean age of 62.74 ± 15.20 (26-86) years in this cohort. About 70% of the patients participated in the study and returned the filled questionnaires. There was a significant overall improvement across all three scores (WFI: p  = 0.001, ODS: p  = 0.001, and BBUS: p  = 0.001). Some individual components in the scoring systems did not improve to patient's satisfaction. No perioperative complication or conversion to an open procedure was reported in this study. Three recurrences were seen in the redo cases. Conclusion  LVMR is a promising way of dealing with rectal prolapse. A careful patient selection, appropriate preoperative workup, and a meticulous surgical technique undoubtedly transform the postoperative outcomes.