Pinprick Method Research Articles

Pin-prick (Medipin) assessment for neuropathy in diabetes: Prospective screening study in primary care

AimsDiabetic patients are at elevated risk of neuropathy; early detection is desirable to minimise the risk of complications. The Medipin pin-prick device was appraised as a screening tool for diabetic neuropathy. MethodsProspective cross-sectional comparative screening study in primary care setting, involving 389 participants with type 2 diabetes mellitus. The Medipin pin-prick method, involving dorsal application on the hallux of both feet, was compared to 10 g monofilament testing. ResultsThe ternary and semi-quantitative approach for scoring Medipin pin-prick sensation give very similar results (Spearman rho 0.67, P < 0.001). A total of 59 % patients had no signs of neuropathy (sharp sensation), 38 % reported impaired sensation (dull sensation), and an absence of sensation occurred in 3 % of patients. For the monofilament dorsal method, the figures were 79 % no neuropathy, 14 % elevated risk, and 7 % neuropathy respectively, and with the monofilament plantar method 87 % of patients had no neuropathy and 13 % did. Correlation analyses showed that taller patients and those with existing neuropathic pain are at very modest increased risk of neuropathy. ConclusionsThe Medipin pin-prick device can identify diabetic neuropathy and detects (first signs of) neuropathy in relatively more patients than 10 g monofilament testing. The differential targeting of nerve types, namely predominant small (Medipin) versus large (monofilament) fibre, likely underpins the difference in outcomes.

First Report of Mango Stem-end Rot Caused by Pestalotiopsis kenyana in Yunnan Province, China.

Stem End Rot (SER) is a devastating post-harvest disease of mango fruits causing severe losses during storage. In 22 July 2023, 31 out of 50 intact mangoes (cv. Sensation) collected from five orchards in Huaping county (26°37'N 101°15') showed typical symptoms of SER after stored for 9 d in room temperature (24-28℃). Initially, small dark brown to black spots appeared around the fruit peduncle, which rapidly expanded through the pulp tissues. The symptomatic mangoes were surface disinfected by 3% NaClO for 30 s after soaking in 75% alcohol for 3 min, and cleaned by sterile water for 3 times. Tissues were cut from the edge of lesions, dried by sterile filter paper, transferred to PDA and cultured at 28 ℃ for 5 d (Tovar-Pedraza et al., 2020). The single-spore isolation method was used to obtain pure culture. Thirty eight isolates presented four distinct kind of morphology on PDA medium. Among them, 11 isolates with same morphology were significantly distinct from common pathogens of SER. The colonies were white and pale yellow on reverse side. Mycelia grew fast and reached the edge of 90 mm Petri dish after cultured for 5d. Pycnidia were black and scattered on the mycelial mats after 15-20 d. Conidia were fusoid, straight to slightly curved, four septa, and brown. Pigmented median cells doliiform, 14.97 - 18.62(16.11 ±0.89)×5.61- 7.28 (6.61±0.51) μm. Apical cell hyaline, subcylindrical; 1-3 tubular transparent apical appendages 12.27 - 16.68 (13.65±3.78)×1.14 - 1.99 (1.59±0.36) μm. Basal cell conical with a truncate base, hyaline, and 1-2 tubulose basal appendages with 2.85 - 7.97 (5.18±1.88)×0.99 - 1.85 (1.38±0.29) μm (n=50). These fungi were described as Pestalotiopsis kenyana. based on morphological characters (Maharachchikumbura et al., 2014) which were different from isolates characterized as other common SER pathogens (Botryosphaeria, Neofusicoccum). Based on morphology, HPSX-4 was selected for further identification. ITS region, tef1-α, β-tub of HPSX-4 were amplified and sequenced (Xun et al., 2023). The sequences were deposited in GenBank (ITS:OR889126, tef1-α:OR913431, β-tub: OR913432). The ITS, tef1-α, β-tub sequence of HPSX-4 showed 100% (525/525)，99.59% (241/242), and 100% (742/742) identity to the P. kenyana CBS442.67 sequences (ITS: NR147549，tef1-α: KM199502, β-tub: KM199395), respectively. HPSX-4 clustered with P. kenyana CBS 442.67 (type strain) based on maximum likelihood method by MEGA 7.0.21(Minh et al., 2013). Pathogenicity test was performed on 12 healthy mangoes (cv. Golek) by placing mycelial plugs around the peduncle and the middle of the fruit by pin-prick method according to Feng et al.(2023). Sterile PDA were used as control (three mangoes). Every inoculated fruit was incubated at 28°C, 95% ± 3% humidity with three replicates for each treatment. The experiment was repeated three times. Typical symptoms of SER were observed. There were no symptoms in the control group. The strain was reisolated and identified as P. kenyana with the method mentioned above which fulfilled Koch's postulates. This is the first report of P. kenyana causing SER disease on Mangifera indica L.. This study expands our understanding of the pathogen range of mango SER which conducive to prevent and control the SER caused by P. kenyana.

First report of Pectobacterium aroidearum causing soft rot on leaf mustard in China.

Leaf mustard (Brassica juncea [L.] Czern. et Coss.) is widely planted in China as an important leaf vegetable. In March 2022, water-soaked and mushy rot symptoms were observed on leaf mustard plants in the field of Zhaotong (27.85°N; 105.05°E), Yunnan, China. The incidence of symptomatic leaf mustard was approximately 10%. The isolation of the causal agent followed the method of Peng et al. (2023). Briefly, infected tissues from four diseased plants were mixed and teased apart, and homogenized by vortex shaking. The bacterial suspension was diluted and spread on nutrient agar (NA). About 10 single colonies exhibiting different colony morphologies were picked and purified separately by successive streaking. A pinprick method was used for pathogenicity tests with an inoculum concentration of 108 CFU/ml (Singh et al. 2013). Among 10 isolates, only strain YKX exhibited soft rot symptoms on detached mustard leaves. In addition, ten two-month-old leaf mustard plants grown in the greenhouse were used for in vivo pathogenicity tests. Briefly, sterilized pins were dipped in the bacterial suspension, and then leaf mustard petioles were pricked with these pins. After inoculation, each plant was kept in a plastic bag for 12 hours to maintain high humidity. As expected, strain YKX caused obvious rot symptoms on eight plants at 1-2 days post-inoculation while the control group including two plants treated with sterile water showed no symptoms. The colonies of strain YKX on NA were white, roughly circular, and convex. For a preliminary identification, total DNA was extracted and used as the template in PCR amplification of 16S rDNA with the universal PCR primer pair 27F/1492R (Weisburg et al. 1991). The quality-filtered DNA sequence (871 bp) showed 100% query coverage and 99.47% identity to the 16S rDNA sequences of type strain Pectobacterium aroidearum SCRI 109T (GenBank: NR_159926) found in the NCBI rRNA/ITS database. Whole-genome sequencing of strain YKX was then performed using the Illumina and Nanopore sequencing platforms by Tsingke Biotechnology Co., Ltd. (Beijing, China). A single contig (GenBank: CP129239) with a length of approximately 4.9 Mb was obtained by de novo hybrid assembly using Unicycler v0.5.0 (Wick et al. 2017). The quality of the genomic data was evaluated by BUSCO v5.4.7 (Manni et al. 2021) against the gammaproteobacteria_odb10 dataset. A BUSCO complete score of 99.5% indicated high assembly quality. The genome sequence of strain YKX was uploaded to the Type Strain Genome Server for a genome-based taxonomic analysis (Meier-Kolthoff et al. 2022). The distance-based phylogeny showed that strain YKX and P. aroidearum L6 (GenBank: CP065044) and P. aroidearum PC1 (GenBank: NC_012917) form a clade. When comparing strain YKX with L6 or PC1, the digital DNA-DNA hybridization value (83.5-83.8%) was above the species delineation threshold (70% for DDH), clearly indicating that strain YKX should be classified as P. aroidearum. Additionally, P. aroidearum was reisolated from inoculated leaves and identified based on morphological similarities and 16S rDNA sequencing, thus fulfilling Koch's postulates. It is worth noting that a previous study reported occurrences of soft rot disease on leaf mustard attributed to Rhizopus microsporus var. chinensis (Wang et al. 2020). To our knowledge, this is the first report of P. aroidearum causing soft rot on leaf mustard in China, which expands the known host range of this pathogen and benefits the control of this disease.

Complete genome sequencing and construction of full-length infectious cDNA clone of papaya ringspot virus-HYD isolate and its efficient in planta expression.

Papaya ringspot virus (PRSV) is a devastating Potyvirus that causes papaya ringspot disease in Carica papaya plantations globally. In this study, the complete genome sequence of a PRSV isolate from Shankarpalli, Telangana, India, was reported and designated as PRSV-HYD (KP743981.1). The genome is a single-stranded positive-sense RNA comprising 10,341 nucleotides. Phylogenetic analysis revealed that PRSV-HYD is closely related to PRSV Pune (Aundh) isolate with 92 and 95% nucleotide and amino acid sequence identity, respectively. To develop infectious cDNA (icDNA), the complete nucleotide sequence of PRSV-HYD was cloned between the right and left borders in the binary vector pCB301 using BglII and XmaI restriction sites. Cauliflower mosaic virus (CaMV) double promoter (35S) was fused at the 5'-end and Avocado sunblotch viroid (ASBVd) ribozyme (RZ) sequence was fused to the 3' end to generate an authentic 3' viral end in the transcribed mRNAs. The icDNA generated was mobilized into the Agrobacterium tumefaciens EHA 105, and the agrobacterial cultures were infiltrated into the natural host C. papaya and a non-host Nicotiana benthamiana plants; both did not show any symptoms. In RT-PCR analysis of RNAs isolated from N. benthamiana, we could detect viral genes as early as 3 days and continued up to 28 days post infiltration. Alternatively, virion particles were purified from agroinfiltrated N. benthamiana plants and introduced into C. papaya by mechanical inoculation as well as by pinprick method. In both cases, we could see visible systemic symptoms similar to that of wild type by 40 days. Additionally, we studied the expression patterns of the genes related to plant defense, transcription factors (TFs), and developmental aspects from both C. papaya and N. benthamiana.

Epidemiological Dynamics of Xanthomonas xonopodis pv. phaseoli in French bean (Phaseolus vulgaris): Unraveling the Factors Governing Bacterial Blight Pathogenesis

The study was conducted during May, 2020 to June, 2021 in the Department of Plant Pathology Laboratory of Dr. Y.S Parmar University of Horticulture and Forestry, Nauni, located in Solan district of Himachal Pradesh, India to study the role of various epidemiological parameters on the development of common bacterial blight of French bean. The diseased samples were collected and the pathogen was isolated and purified on Luria Bertani (LB) agar plates. The pathogenicity test was performed on the potted plants as well as on the detached leaves using carborundum abrasion and pin prick method of inoculation. The incubation period on potted plants using carborundum abrasion and pin prick method revealed the initial symptoms after 117.60 h and 120.30 h, respectively, while on detached leaves they were visible after 72.30 h and 84.50 h of inoculation, respectively. Further to study the effect of different epidemiological factors on disease development, the leaves inoculated using carborundum abrasion were subjected to four temperature levels, five relative humidity levels and five different durations of leaf wetness under controlled conditions in relative humidity cum temperature control cabinet. The observations were recorded at different intervals and the data revealed that the pathogen caused severe symptoms on the leaves with high apparent rate of infection under 30°C temperature, 100% RH and a maximum of 24 h of leaf wetness. It was observed that the severity of disease decreased as the temperature raised from 30°C and increased with the rise in RH levels and leaf wetness duration.

In-vivo Screening of Cherry Tomato [Solanum lycopersicum L. var. cerasiforme (Dunnal) A. Gray] Genotypes and Hybrids against Fusarium Wilt in Arunachal Pradesh, India

Cherry tomato [Solanum lycopersicum L. var. cerasiforme (Dunnal) A. Gray] is becoming popular among various tribes of Arunachal Pradesh due to its unique taste, flavor and appearance. Owing high rainfall and high humidity, successful cultivation of cherry tomato is becoming restricted in this state due to various biotic factors like infection of wilt complexes as well as pests like root-knot nematodes. The aim of the present investigation was to evaluate eighteen cherry tomato genotypes (nine) and hybrids (nine) against Fungal wilt caused by Fusarium oxysporum f. sp. lycopersici resistance in the Pasighat condition of Arunachal Pradesh, India. For screening purpose, artificial pathogen inoculation method was applied through pin-prick method in the 35 days old cherry tomato seedlings that were grown in pot culture containing sterile soil and data for the pathogen infection was recorded by following Completely Randomized Design (CRD) in 60 days and 120 days after pathogen inoculation. Among 18 genotypes and hybrids, G4xG6 (DI:0.00%, PDI:0.00%), G9 (DI:0.00%, PDI:0.00%) and G5 (DI:0.00%, PDI:0.00%) showed highest resistance towards Fusarium wilt, while genotype G3 (DI:65.56%, PDI:63.60%) exhibited susceptible system. The disease incidence was ranged from 0.00%-65.56%. The use of resistant genotypes and hybrids to manage the population of fungus is a very cost effective method and can be exploited commercially in breeding programmes and for vegetable grafting.

Comparative Study of Effect of Dexmedetomidine and Fentanyl as Epidural Adjuvants in Abdominal Hysterectomies Using Combined Spinal Epidural Technique - A Randomised Prospective Study

Background: Regional anesthesia is defined as loss of sensation in a body produced by administration of anaesthetic agent to the nerves supplying that region. Regional anesthesia whether by spinal, epidural, peripheral nerve blocks offer a number of advantages. Bupivacaine has become standard agent for intrathecal anesthesia. A number of adjuvants can be added to prolong the duration of analgesia. In our study we are comparing fentanyl &amp; dexamedetomidine as an epidural adjuvant to spinal bupivacaine in abdominal hysterectomies. Materials and Methods: A total of 60 patients scheduled for abdominal hysterectomy under CSE were enrolled for this prospective randomized study which was conducted in hospital from November 2019 to October 2020. This work has been granted ethical committee approval. Results: Two groups comprising of 30 patients each were taken randomly. Haemodynamic parameters of heart rate, blood pressure, spo2, were monitored and recorded every 5 min for first 20 minutes and then every 10 minutes till the end of surgery and thereafter postoperatively. The sensory block was assessed by bilateral pin prick method and motor blockade according to Modified Bromage Scale. Duration of anesthesia was taken as time period till VAS of 4 was recorded. Conclusion: In our study we concluded that dexmedetomidine is better adjuvant as compared to fentanyl in combined spinal epidural anaesthesia with intrathecal bupivacaine.

First report of stem-end rot caused by Botryosphaeria fabicerciana on mango fruits in China.

Mango (Mangifera indica L.), belongs to the family Anacardiacea, and is one of the most popular tropical fruits in the world. Stem-end rot is a major postharvest disease of mango fruit, causing severe losses during storage in China (Chen et al., 2015). In July 2021, the mango fruits harvested from Baise Municipal National Agricultural Science and Technology Park (23.683568 N, 106.986325 E) of Guangxi province in China developed stem-end rot during storage. The disease incidence reached ca. 8.3%. The initial symptoms appeared as light brown lesions surrounding the peduncle, which quickly expanded becoming large dark-brown lesions. Small pieces of epidermis (5 mm × 5 mm) from 8 typical diseased friuts were cut from the edges of lesions surface-sterilized with 2% sodium hypochlorite and rinsed with sterile distilled water. The tissue was plated on potato dextrose agar (PDA) and incubated at 28 ℃ in the dark for 3 days. Fifteen, similarcolonies were isolated from the symptomatic tissue. The representative isolates DF-1, DF-2 and DF-3 were selected for morphological characterization, molecular identification, and pathogenicity testing. The colonies were circular with fluffy aerial mycelium, initially white turning to smoke-gray from the center in upper side and greenish black in reverse side, covering the 90 mm diameter Petri dish after 4 days of incubationon PDAat 28 ℃in dark. Pycnidia were produced on the surface of the colony after 30 days. Conidia were fusiform, aseptate, hyaline, thin-walled with granular contents, apex sub-obtuse, base subtruncate to bluntly rounded, 14.0-20.3 (16.8±1.6) μm × 3.1-7.2 (5.1±0.9) μm (n=50). The sexual stage was absent. Based on morphology, isolates were preliminarilyidentified asBotryosphaeriaspeices.To accurately identify the pathogen, genomic DNA was extracted from the mycelium of the three isolates DF-1, DF-2 and DF-3. The internal transcribed spacer of rDNA region (ITS), elongation factor 1-alpha (EF-1α) and beta-tubulin gene (TUB) genes were amplified using primers ITS1/ITS4, EF1-728F/EF1-986R and Bt2a/Bt2b, respectively (Slippers et al., 2004). The nucleotide sequences were all deposited in GenBank (ITS: OP729176-OP729178 EF-1α: OP758194-OP758196 and TUB: OP758197-OP758199). Based on the BLASTn analysis, the ITS, EF1-α and TUB sequences of three isolates were 100%, 99% and 99% similar to the Botryosphaeria fabicerciana MFLUCC 10-0098 sequences (ITS: JX646789, EF-1α: JX646854 and TUB: JX646839). Multi-locus phylogenetic analyses (ITS, EF-1α and TUB) showed that the isolate DF-1, DF-2 and DF-3 were clustered within Botryosphaeria fabicerciana clade based on the maximum likelihood , Bayesian inference, and maximum parsimony methods. The pathogenicity test was performed by placing discs mycelium around the peduncle of mature mango fruits by pin-prick method. Each treatment carried out with 12 fruits. The inoculated fruits were placed in plastic boxes at 28 ℃ with three replicates. Three days after inoculation, typical symptoms of stem-end rot were observed. The control fruits were inoculated with sterile PDA discs, and remained symptomless. The same fungus was re-isolated from the symptomatic tissue to complete Koch's postulate. Botryosphaeria fabicerciana (basionym: Fusicoccum fabicercianum) was first reported as pathogen causing senescent twig of Eucalyptus spp. in China (Chen et al., 2011; Phillips et al., 2013). To our knowledge, this is the first report of Botryosphaeria fabicerciana causing stem-end rot of Mangifera indica in China.

Genetic analysis and identification of SSR marker linked to Phomopsis blight resistance in eggplant (Solanum melongena L.)

AbstractThe present investigation was carried out to decipher inheritance of resistance and to identify linked SSR markers for Phomopsis blight resistance in eggplant. An F2 population comprising 161 plants was developed from the cross of Pusa Kranti and BR‐40‐7. Genetic analysis was carried out using Chi square test. Artificial inoculation of fruits was carried out using pin prick method, and scoring was done as per the standard scoring scale. The F2 plants segregated into 92 susceptible (77—highly susceptible, 15—susceptible): 69 resistant (17—highly resistant, 27—resistant, 25—moderately resistant) suggesting complimentary epistasis with ratio of 9:7. To identify the putatively linked markers to resistance gene, parental polymorphic markers were subjected to bulk segregant analysis (BSA), and two markers (emk03O04 and emf11A03) could differentiate resistant and susceptible bulk and co‐segregated with resistance gene. The genetic distance between the identified markers was found to be 18.12 cM using QTL IciMapping V3.2 software depicting two new QTLs on chromosome number 6. The identified QTLs have great significant importance in marker assisted breeding programme.

Effect Of Perineural Administration Of Dexmedetomidine With Bupivacaine Versus Bupivacaine Alone In Ultrasound Guided Supraclavicular Block For Upper Extremity Orthopaedic Operations.

During procedures on the upper limbs, the brachial plexus block is usually advised. To increase the length of the block, many medicines have been utilized as adjuvants. The purpose of this study was to compare the effects of dexmedetomidine plus bupivacaine against bupivacaine alone on the onset and duration of the sensory and motor block and the duration of analgesia in the supraclavicular block during upper extremity orthopaedic surgery. Sixty individuals qualified for orthopaedic operations on the upper extremities, ranging in age from 20 to 60 years, participated in this prospective, randomized investigation. The modified Bromage scale and the pinprick method were used to assess the sensory and motor block. Using a visual analogue pain scale, the postoperative pain was evaluated at 0, 6, 12 and 24 hours after surgery. In patients receiving only bupivacaine, the mean onset time of sensory and motor block was 32.84 minutes and 26.67 minutes respectively; while in those receiving bupivacaine along with dexmedetomidine, it was 23.38 minutes and 14.81 minutes (p<0.005). In the intervention group (bupivacaine and dexmedetomidine), the period between the first request for analgesia and the duration period of sensory and motor block were both longer (p<0.005). The intervention group experienced less postoperative discomfort for 24 hours (p<0.05). Dexmedetomidine added to bupivacaine perineurally prolonged both numbness and immobility while shortening the time it took for sensory and motor blocks to begin. Moreover, dexmedetomidine considerably decreased postoperative pain when combined with bupivacaine for supraclavicular blocks.

Elucidation of Physio-Biochemical Changes in Citrus spp. Incited by Xanthomonas citri pv. citri

India is one of the leading citrus−producing countries, next to China and Brazil. Despite the sizeable production, especially of lemons and limes, India contributes meagerly to the world’s citrus market. Of the various factors responsible for the restricted quantum of citrus export, citrus canker (Xanthomonas citri pv. citri) is one of the leading serious causes and directly impacts the appearance of the fruits. Considering the extent of damage, the present study aimed to understand the impact of Xanthomonas citri pv. citri (Xcc) on the physio−biochemical responses in contrasting Citrus spp. Two genotypes, each of three citrus species, namely Citrus aurantifolia (Pusa Udit and ALC−35), C. limon (Kagzi Kalan and Konkan Seedless), and C. paradisi (Redblush and Marsh Seedless), were artificially inoculated with Xcc (108 to 109 cfu/ mL) by the pinprick method. The physio−biochemical changes in the host were evaluated after 48 h post inoculation (hpi). The chlorophyll content (total, a, and b) degradation and reduction in leaf gas exchange parameters, such as photosynthetic rate (A), transpiration rate (E), stomatal conductance (gs), and intrinsic water−use efficiency (iWUE), were measured to a greater extent in susceptible than resistant genotypes. The microscopic observations also evidenced higher stomatal density with larger stomatal areas in susceptible genotypes, favoring the easier penetration of Xcc in host tissues than resistant species or genotypes. The higher activities of various antioxidant enzymes, viz., superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and glutathione reductase (GR), the contents of soluble protein, and phenolics were measured in resistant genotypes in contrast to susceptible genotypes. The activities of phenyl ammonia lyase (PAL) and polyphenol oxidase (PPO) were also higher in resistant genotypes, whilst the levels of ROS (superoxide radical and hydrogen peroxide) production were enhanced in susceptible genotypes. Based on the host–pathogen interaction, the order of resistance in a descending manner was found as Kagzi Kalan, Marsh Seedless, Redblush, Konkan Seedless, Pusa Udit, and ALC−35. Further, the results will enhance the understanding of the pathogen mechanism during pathogenesis in resistant and susceptible Citrus species.

Effect of dexmedetomidine and fentanyl on hemodynamic changes and block profile following spinal anesthesia with ropivacaine among patients with femoral fractures undergoing lower limb surgery.

The aim of this study was to compare the effect of dexmedetomidine and fentanyl on hemodynamic changes and block characteristics following spinal anesthesia with ropivacaine among patients with femoral fractures undergoing lower limb surgery. In this double-blind clinical trial, 64 patients who were candidates for lower limb surgery. Patients were divided into two groups based on the block pattern. In the first group, dexmedetomidine was prescribed. In the second group, fentanyl with ropivacaine was prescribed. Sensory and motor blocks at or above the T8 dermatome in each group were measured. Furthermore, the sensory block was evaluated every 1 minute after anesthesia with a needle (pin prick method) and also the motor block was evaluated every 5 minutes by the bromage scale. There was a statistically significant difference between the two groups in terms of the time for achieving sensory block to T8 or higher dermatome (p = 0.0001). The time elapsed until the onset of motor block was shorter in the dexmedetomidine group, and dexmedetomidine had a shorter time for achieving sensory block to T8 or higher dermatome than fentanyl. A statistically significant difference was found in terms of the time elapsed until the motor block and the time for achieving sensory block to the T8 dermatome or higher (p <0.05). The time elapsed until the onset of motor block was shorter in the dexmedetomidine group, and dexmedetomidine had a shorter time for achieving sensory block to T8 or higher dermatome than fentanyl. Our findings revealed a statistically significant difference in terms of the duration of sensory block for reaching the T12 to L1 dermatome and the duration of obtaining bromide scores 0 and 1 (p = 0.0001). The time for achieving sensory block to dermatome T12 to L1 and the time of obtaining bromage scales of 0 and 1 were longer in dexmedetomidine group (p = 0.0001). Pain in dexmedetomidine group was less than fentanyl group in 2 to 8 hours after surgery (p <0.05). The duration of analgesia was longer in the dexmedetomidine group (p = 0.001). In summary, it can be suggested that adding dexmedetomidine to the anesthetic ropivacaine may be beneficial.

First report of Pseudomonas palleroniana causing potato soft rot in China.

Slimy potato (Solanum tuberosum) tubers with a foul-smelling odor were obtained from a potato field during harvest in Huize county (25.94°N; 103.40°E), Yunnan province, China in August 2021. The incidence of symptomatic potato tubers was approximately 5% while no symptoms were observed on potato stems. To isolate the causal agent, potato tubers were surface-disinfected and infected tissues were mashed in a 15 ml sterile centrifuge tube with 2 ml sterile distilled water. After a series of dilutions, a volume of 200 μl bacterial suspension from each dilution was spread on nutrient agar (NA) medium and incubated at 28 °C for 48 h. Single colonies randomly selected from NA plates were then picked and separately subcultured in nutrient broth (NB) medium. Pure cultures were acquired by successive streaking on NA medium. The colonies with irregular shapes were white and opaque (Fig. S1A). Among all the isolates, strain Q1 exhibited pectinolytic activity on inoculated potato tubers using a pin prick method with an inoculum concentration of 108 CFU/ml while no symptoms appeared for the control group using sterile water (Fig. S1B). However, no obvious pits were observed for strain Q1 when tested on crystal violet pectin (CVP) medium. To further identify the strain Q1, total DNA was extracted using the TaKaRa MiniBEST Bacteria Genomic DNA Extraction Kit. PCR amplification of 16S rDNA of strain Q1 was performed using the universal PCR primer pair 27F/1492R and followed by Sanger sequencing. The BLASTn analysis of the sequence (NCBI accession number ON631256) based on NCBI rRNA/ITS databases revealed 100% query coverage and 99.78% identity to the 16S rDNA sequences of type strain Pseudomonas tolaasii ATCC 33618 and Pseudomonas palleroniana CFBP 4389, respectively. Whole-genome sequencing of strain Q1 was then performed using the Illumina and Nanopore sequencing platform. A single contig (NCBI accession number CP092411) with a length of approx. 6.2 MB was obtained by de novo assembly using Raven (v1.5.1) (Vaser and Šikić 2021) and Pilon (v1.24) (Walker et al. 2014). The completeness and redundancy of this assembly was evaluated by BUSCO (v5.2.2) (Manni et al. 2021), and a score of 100% completeness indicated a high quality of the assembly. The genome sequence of strain Q1 was uploaded to the Type Strain Genome Server (TYGS) for a whole genome-based taxonomic analysis (Meier-Kolthoff and Göker 2019). The distance-based phylogeny revealed that strain Q1 is well clustered together with Pseudomonas palleroniana (Fig. S1C). Furthermore, a digital DNA-DNA hybridization (dDDH) value (86.9%) between strain Q1 and P. palleroniana was above the species boundary (≥70%), indicating that strain Q1 should be classified as P. palleroniana. In addition, P. palleroniana was re-isolated from potato tubers inoculated using a pin prick method with an inoculum concentration of 108 CFU/ml and was identified by morphological similarities and 16s rDNA sequencing described above, thus fulfilling Koch's postulates. Furthermore, aerial stem rot symptoms were observed after 3-5 days post inoculation of strain Q1 with a concentration of 108 CFU/ml on stems of young potato plants using a pin prick method (Fig. S1D). Notably, a couple of Pseudomonas spp. have been reported to cause soft rot in carrot (Godfrey and Marshall 2002) and melon (Zhang et al. 2016). To our knowledge, this is the first report of P. palleroniana causing soft rot on potato tuber in China.

Prospective Study Evaluating the Association between Time since Scorpion Bite and Spinal Anaesthesia Outcomes

Introduction: The drug mainly used in Spinal Anaesthesia (SA) is Local Anaesthetic (LA) agents. It acts via sodium channel by reversibly inhibiting the nerve transmission. Scorpion venom causes mutation in receptors of sodium channel that leads to LA resistance which results in failed SA. Although, there are few studies and many case reports, which shows direct association of scorpion bite with failed SA, but association of duration since, scorpion bite on outcome of spinal anaesthesia is still not known. Aim: To evaluate the association between time since scorpion bite and multiple bites, an outcome of SA on patients posted for elective lower abdominal and lower limb orthopaedic surgeries. Materials and Methods: This monocentric, prospective, single blind observational study was conducted in the Department of Anaesthesia at Rajarshi Dashrath Autonomous State Medical College (RDASMC), Ayodhya, Uttar Pradesh, India. The duration of the study was two years, from December 2019 to December 2021. A total of 33 patients of either gender or age, between 18-70 years were taken and divided them into three groups of 11 each. Groups were divided on the basis of history of duration since scorpion bite/sting. Group A included patients with history of scorpion bite being less than eight months, group B from eight-14 months and group C more than 14 months. SA was provided with 15 mg 0.5% hyperbaric bupivacaine. The pin prick method and Bromage scale were used to detect the onst of sensory (T1), onset of motor (T2), peak of sensory (T3) and peak of motor block (T4). General Anaesthesia (GA) was given if the SA was found inadequate or failed. The mean±Standard Deviation (SD), Student’s t-test and Chi-square test were used for statistical analysis of data. Results: The mean age of the study participants of group A was 49.27 years, group B was 50.27 years and group C was 52.18 years. Demographically, all the three groups were comparable. The authors have found that, block failed completely in group A. There was statistically significant increase of T1, T2, T3 and T4 in group B than group C (p&lt;0.05). Statistically significant difference were not found between single scorpion bite and multiple scorpion bite in group B and group C in terms of SA characteristics T1, T2, T3 and T4 (p&gt;0.05). Conclusion: The present study concludes that, the duration of scorpion bite is a significant factor on the effect of SA in terms of sensory and motor block, but number of stings does not determine the sensory and motor characteristics of SA. The SA should not be given if, the history of scorpion bite is recent (less than eight months).

Salvianolic acid B combined with bone marrow mesenchymal stem cells piggybacked on HAMA hydrogel re-transplantation improves intervertebral disc degeneration.

Cell-based tissue engineering approaches have emerged as a realistic alternative for regenerative disc tissue repair. The multidirectional differentiation potential of bone marrow mesenchymal stem cells (BMSCs) to treat disc degeneration intervertebral disc degeneration has also become a viable option. We used 1% HAMA hydrogel as a carrier and co-encapsulated BMSCs and Salvianolic acid B (SalB) into the hydrogel to reduce the apoptosis of the transplanted cells. The protective effect of SalB on BMSCs was first verified in vitro using the CCK8 method, flow cytometry, and Western-Blotting, and the physical properties and biocompatibility of HAMA hydrogels were verified in vitro. The rat model was then established using the pinprick method and taken at 4 and 8 W, to examine the extent of disc degeneration by histology and immunohistochemistry, respectively. It was found that SalB could effectively reduce the apoptosis of BMSCs in vitro by activating the JAK2-STAT3 pathway. 1% HAMA hydrogels had larger pore size and better water retention, and the percentage of cell survival within the hydrogels was significantly higher after the addition of SalB to the HAMA hydrogels. In the in vivo setting, the HAMA + SalB + BMSCs group had a more pronounced delaying effect on the progression of disc degeneration compared to the other treatment groups. The method used in this study to encapsulate protective drugs with stem cells in a hydrogel for injection into the lesion has potential research value in the field of regenerative medicine.

Identification of new resistant varieties to Phomopsis fruit rot of eggplant (Solanum melongena)

Eggplant (Solanum melongena L.) is one of the supreme members of the solanaceous group which is widely grown in India. It is considered as the single and economic host of the fungus Diaporthe vexans causing fruit rot, stem canker and blight disease. The present study was carried at the vegetable research farm of ICAR-Indian Agricultural Research Institute, New Delhi during 2019–20 and 2020–21 to screen the eggplant lines (18 cv.) for resistance against this pathogen. The pathogen was isolated from the infected fruits and confirmed by morphological and sequencing information. Artificial inoculation using pin prick method revealed the presence of variability in the lines for disease. The studied lines were categorized based on Per cent disease index (PDI) and the highest PDI value of 80–100% recorded in cultivars Pusa Shyamla, Pusa Kranti, DBL-186, DBL-187, Muktokeshi and DBSR-52 and thus categorized as highly susceptible. The cultivars NDB-25, Kashi Sandesh, Kalo Solia, DBL-175 and Debjhuri Hazari were reported to be susceptible with PDI value of 60%. In the present study, first time we have report a number of highly resistant lines, viz. BR 112, Swarna Mani, BR 40-7 and IC-112992 with 0% PDI as novel source of resistance against P. vexans. These resistant lines have significant importance in large scale cultivation as resistant varieties by farmers. They can also be further utilized as a parent in population development for the identification of gene (s)/ QTL (s) associated with the resistance.

Emergency decompressive craniotomy with scalp block and closed-loop TCI sedation for acute ischemic stroke in a patient with incidental diagnosed large left ventricular mass, presenting with severe LV dysfunction.

Dear Editor, An average built 35-year-old male presented to the emergency-department with a history of a sudden onset of left-sided weakness and bi-frontal headache, with a chronic history of ischemic cardiomyopathy, and acute myocardial infarction one year back, with current functional status New York Heart Association classification (NYHA) class III. Patient underwent thrombolytic therapy with 0.9 mg/kg alteplase (r-TPA) over 60 minutes after evaluating risk factors during the precious golden hour period. Over the next 12 hours, the GCS deteriorated. There was cerebral swelling with a midline shift of 8 to 12 mm, although the patient was not intubated or ventilated. In a repeat scan and the neurosurgeon planned to proceed with emergency decompressive craniotomy. Complete hemogram was normal, electrocardiogram revealed sinus tachycardia with left anterior hemiblock. Routine trans-thoracic echocardiogram suggested global left ventricular (LV) dysfunction with an ejection fraction of 20%, large left ventricular mass (query myxoma or thrombus attached to LV apex measuring 44 mm × 36 mm), and grade III mitral regurgitation with eccentric jet. [Figure 1] Computerized tomography showed Acute Ischemic Stroke (AIS) in the right middle cerebral artery territory. After explaining due appropriate risks and benefits of surgery to the patient relatives and interdisciplinary consensus, the team decided to proceed with craniotomy under Monitored Anesthesia Care using scalp block and manual closed-loop TCI Propofol sedation under spectral entropy monitoring.Figure 1: Echocardiography (ECHO) Revealing Large Left Ventricular Mass Attached LV Apex Measuring 44 Mm × 36 Mm (A) Parasternal Long Axis View, (B) Apical Four Chamber ViewBilateral scalp anesthesia was given using 24-gauge short bevel needle with 40 ml 0.2% Ropivacaine. After assessing for adequate block by pin prick method, entropy leads were attached and secured with 1-inch plaster tape. TCI sedation with Propofol was started with the initial effect site concentration of 1 microgram/ml, later escalated to 3 microgram/ml to maintain spectral entropy between 60-80 and hemodynamic parameters. Decompressive craniotomy was completed in 2 hours 10 minutes, with no complications; patient recovery was rapid and was obeying verbal comments when effect-site concentration reached 0.5 microgram/ml. After this emergency surgery, the patient was operated for his cardiac mass after 9 months and it was found to be thrombus on histopathological diagnosis. Thrombus, tumor, and vegetation are the most common causes of intracardiac mass and must be distinguished from normal variants such as papillary muscles, false tendons, sigmoid septum, and apical trabeculation.[1] Our patient was at high risk with associated large left ventricular mass, severe left ventricular dysfunction, and ischemic cardiomyopathy and grade III mitral regurgitation for emergency craniotomy. Anesthetic management in this clinical scenario could be associated with risk of complications ranging from peri-operative myocardial infarction, pulmonary edema, LV outlet obstruction and sudden cardiac arrest, new thrombo-embolic episodes, and hemorrhagic changes in infraction.[2] Scalp block with TCI Propofol sedation prevents hemodynamic perturbances associated with general anesthesia with intubation and cardio depressant effects of anesthetic agents. Manual closed loop TCI Propofol sedation with deep anesthesia control offers optimal titration with stable hemodynamic sedation and eliminates the need for unintended protection of the airway.[3] Considering the risk and benefit ratio, craniotomy with closed loop TCI Propofol sedation outweighed the risks of general anesthesia in our patient. Though awake craniotomy with TCI sedation is safe but has a few limitations like difficulty to alter ventilation in case of intraoperative cerebral edema and airway securement may be difficult in case requirement occur intraoperatively due to positioning of patient. In spite of appropriate anticoagulation, a thrombus traveling across the cardiac cycle has a high potential to embolize. It is imperative LV masses should be surgically removed as early as possible, owing to risk of recurrent thrombo-embolic phenomenon. We conclude that craniotomy under scalp block may be performed even in an emergency craniotomy in patients with high peri-operative risk. Declaration of patient consent The authors certify that they have obtained all appropriate patient consent forms. In the form the patient(s) has/have given his/her/their consent for his/her/their images and other clinical information to be reported in the journal. The patients understand that their names and initials will not be published and due efforts will be made to conceal their identity, but anonymity cannot be guaranteed. Financial support and sponsorship Nil. Conflicts of interest There are no conflicts of interest.

A COMPARATIVE STUDY BETWEEN 0.5% BUPIVACAINE (HEAVY) AND 2- CHLOROPROCAINE (PRESERVATIVE FREE) FOR SPINAL ANAESTHESIA IN ELECTIVE SHORT SURGICAL PROCEDURES

Background: Spinal anaesthesia is the preferred anaesthetic technique in lower abdominal surgeries but in case of short surgeries (&lt; 60 min), appropriate choice of local anaesthetic is of prime importance to prevent problems like prolonged motor block, delayed ambulation and delayed micturition postoperatively. Preservative free 2-Chloroprocaine has recently come up as a good alternative as compared to commonly used local anaesthetics. Aim: The aim of this study was to compare the efcacy between 1% 2-Chloroprocaine and 0.5% Bupivacaine (heavy) in terms of early block resolution and early ambulation. Material And Methods: This prospective, randomised, double blinded, comparative study was conducted in the Department of Anaesthesiology, North Bengal Medical College and Hospital. One hundred and ten (110) patients of ASA physical status I or II and age of 18-60 yrs undergoing lower abdominal surgeries under spinal anaesthesia were chosen and randomly divided into two groups, Group A and Group B. Group A received 40 mg (4ml) of intrathecal 1% preservative free 2-Chloroprocaine and Group B received 12 mg (2.4ml) of intrathecal 0.5% Bupivacaine (heavy). After administration of intrathecal drugs, vital parameters were monitored and any intraoperative complications recorded. Onset and height of sensory block were monitored by pinprick method and degree of motor block was monitored by modied Bromage Scale – (0-3). Time to reach readiness for surgery, duration of sensory block and motor block, time to ambulation and micturition, any supplementation required and any complications were recorded. The data were analysed using appropriate statistical tests. Results: The patients in Group A had a lower duration of motor block in comparision to those in Group B [81 ± 11 min vs 142 ± 29 min (P&lt;0.001)]. The duration of sensory block was also signicantly lower in Group A as compared to Group B [100 ± 14 min vs 188 ± 39 min (P&lt;0.001)]. Also patients in Group A had lower time to ambulation than those in Group B[142 ± 24 min vs 280 ± 51 min (P&lt;0.001)] as well as lower time to micturition as compared to those in Group B[204 ± 37 min vs 365 ± 57 min (P&lt;0.001)]. Conclusion: For spinal anaesthesia in lower abdominal short surgical procedures, intrathecal 2-Chloroprocaine produces a satisfactory surgical block and is a better drug in comparison to hyperbaric bupivacaine with respect to faster block resolution and ambulation.

Abstract No.: ABS1604: A comparative study of bupivacaine versus ropivacaine in preoperative fascia iliaca compartment block in patients posted for proximal femur fracture surgeries under spinal anaesthesia

Background and Aims:Positioning for spinal anaesthesia is challenging in patients with femur/hip fractures as it causes severe pain/distress. Peripheral nerve blocks are safe adjuncts without side-effects of systemic analgesics. This study was conducted to compare the efficacy between bupivacaine and ropivacaine in ultrasound-guided fascia iliaca compartment block (FICB) for positioning of spinal anaesthesia.Methods:70 patients( 35 in each group) of American Society of Anesthesiologists physical class I-III, either sex, aged between 18-70 years, posted for proximal femur surgeries were randomly allocated in two groups. All patients in group B received 30ml of 0.25%bupivacaine and in groupR 30ml of 0.25%ropivacaine was administered for FICB.. Onset of sensory block was noted every two minutes using pinprick method. Fifteen minutes later spinal anaesthesia was performed with 0.5% hyperbaric bupivacaine in sitting position. Analgesia during positioning was assessed. The quality of patient positioning for spinal anaesthesia was recorded. Vitals were monitored before(baseline) and after block for every 5minutes until 15minutes. The time for first rescue analgesic request and 24h opioid consumption was noted.Results:Mean onset of FICB in group B was 6.17±1.403 minutes and group R was 10.51±1.772 minutes which was statistically significant (p value <0.001). There was no significant difference in visual analogue scale(VAS) during positioning and quality of positioning in both groups. Time to first rescue analgesia in group B was 409.71±85.146 minutes and groupR was 445.71±77.736 minutes (p value 0.0069) but the difference was not statistically significant. There was no significant difference in 24hour opioid consumption among the two groups.Conclusion:FICB provides analgesia which helps in positioning of the patient for spinal anaesthesia with bupivacaine having faster onset of action compared to ropivacaine but provides comparable duration of analgesia and 24 hour opioid consumption.

BIOCHEMICAL CHARACTERIZATION AND EFFECT OF BACTERIAL BLIGHT ON MICRO AND MACRO NUTRIENT CONCENTRATION IN POMEGRANATE

Bacterial blight is a serious disease of pomegranate. The disease affects the yield and quality of pomegranate fruits. In this present work, we investigated the biochemical characterization and role of micro and macro nutrients in diseased and healthy leaves of pomegranate. Pathogen was isolated infected leaves, twigs and fruits and grown on different culture media. Different biochemical tests (Gram staining, KOH, catalase, oxidase, starch hydrolysis, gelatin liquefaction) were performed to identify the bacteria. All these tests confirm the presence of Xanthomonas axonopodis pv punicae. Pathogenicity was done in the greenhouse using three different methods (Pin-pricked, infiltration and spray method) and symptoms appeared in 8 days; and infiltration method showed early symptoms compared to the spray and pin prick method. Ionic contents (Nitrogen, Potassium, Phosphorus, Magnesium, Calcium, Iron, Boron, Zinc and Copper) are reduced by using these nutrients for the growth, multiplication and survival of pathogenic bacteria. Proper application of these nutrients helps the host plant to improve its physiological and biochemical development and resistance to bacterial blight disease in pomegranate.