Assessment of radiation-induced sterility, longevity, and reproductive characteristics of the Aedes aegypti (Diptera: Culicidae) strain of Swat: A step towards the implementation of SIT in Pakistan.

BACKGROUND OBJECTIVES: Biological approaches for vector mosquito control such as sterile insect technique (SIT) requires sterilization of male mosquitoes through specific radiation doses for sterility induction in males. Under the SIT program, the males used must be compatible with the wild males in vigor, flight and selection of mate. Much of this potential is determined by the diet provided in the larval stages and optimizing the irradiation doses that cause complete sterility but have very minimal effect on the lives of irradiated males. The current study is designed to evaluate gamma radiation doses for inducing sterility in male Aedes aegypti mosquitoes with a genomic background from Pakistan (Swat district) and to assess the effects of radiation exposure on the competency of irradiated males and other life traits of irradiated mosquitoes. METHODS: Sterilization of 17-hour-old male pupae (groups of 50 pupa/cup in three replicates) of the Ae. aegypti Swat strain was conducted using radiation doses of 30, 60, 70, 90, and 105 gray (Gy) from (Co60) at the Nuclear Institute for Food and Agriculture (NIFA) in Peshawar, Pakistan. Post-irradiated pupae were released in adult cages. Virgin females of the same cohort were released (1:1) to mate with the irradiated males. Mortality, longevity, mating competitiveness of males, and female fecundity were recorded. RESULTS: An average of 71 eggs per female was recorded in control with 86% hatch rate. Individual females mated with a 60 Gy treated males produced 60 ± 0.6 eggs per female with 17% hatch rate, and those mated with 70 Gy males produced 42 ± 0.01 eggs with a nil hatch rate, whereas females mated with males treated with ≥70 Gy doses did not reproduce to next generation. Females in groups of 50 mated with 60, 70 Gy treated males (equal pairs), produced 369±1.3 and 98±0.01 eggs with 15% and zero hatch rate. Significant dose dependent reduction in longevity was observed for >30 Gy doses. The matting competence of irradiated males was about half that of un-irradiated males. INTERPRETATION CONCLUSION: Aedes aegypti with a Pakistani genomic background treated with a ≥70 Gy dose of gamma radiation induced complete sterility in males and provided the first- step foundation for SIT application in Pakistan. Further extensive studies are required to optimize the SIT techniques so that fully sterile males with very minor quality changes can be produced on large scales for field trials.

Evaluation of Insect Growth Regulators (IGRs) as biological pesticides for control of Aedes aegypti mosquitoes.

BACKGROUND OBJECTIVES: Insect growth regulators (IGRs) are biological hormone analogue or mimics used as pesticides to inhibit the growth of larva during their molting and skin shedding. This study aimed to test the effect of IGRs on the eggs hatching and post-hatching inhibition of Aedes mosquitoes and understanding its effect in the mosquito breeding habitats for reduction in adult emergence. METHODS: Experiments on the evaluation of three insect growth regulators (IGRs) for the control of different stages of Aedes aegypti was carried out during 2020-21. Each experiment consisted of four treatments viz., Pyriproxyfen, Novaluron, and Larvicol at 1.0 ppm and distilled water as a control. All experiments were carried out in completely randomized design (CRD) except eggs which were carried out in factorial design each with three replications. RESULTS: All tested IGRs performed better in affecting eggs, larval and pupal stages of Ae. aegypti. Highest eggs hatching inhibition (80%) of fresh eggs occurred in Pyriproxyfen followed by Novaluron (66%) and lowest in Larvicol (62%). Eggs hatch inhibition of embryonated eggs was lower than fresh eggs. Pyriproxyfen caused 69%, Novaluron 59% and Larvicol 39% eggs hatch inhibition of embryonated eggs. Both Pyriproxyfen and Novaluron performed better in causing 98-100% larval mortality followed by Larvicol (39%). Larval development to pupal stage was completely prevented by both Pyriproxyfen and Novaluron. Although Larvicol resulted in lowest eggs hatch and larval inhibition but prevented pupae to emerge as adults. Results further showed 70-89% mortality of 3rd instar larvae of Ae. aegypti when exposed to Pyriproxyfen and Novaluron solutions after 30 days storage at lab. temperature (27±2°C), RH 70±5. INTERPRETATION CONCLUSION: None of the IGRs was more effective at the pupal stage but showed carry-on activity of growth inhibition and mortality of the successive stages of development when used against eggs stages. Therefore, we recommend early application of IGRs at mosquito habitats during the beginning and onset of the season when very early stages of mosquitoes are available in the field.

Cutting Edge Developments in Spine Surgery at the University of Missouri.

The treatment of spinal pathologies has evolved significantly from the times of Hippocrates and Galen to the current era. This evolution has led to the development of cutting-edge technologies to improve surgical techniques and patient outcomes. The University of Missouri Health System is a high-volume, tertiary care academic medical center that serves a large catchment area in central Missouri and beyond. The Department of Neurosurgery has sought to integrate the best available technologies to serve their spine patients. These technological advancements include intra-operative image guidance, robotic spine surgery, minimally invasive techniques, motion preservation surgery, and interdisciplinary care of metastatic disease to the spine. These advances have resulted in safer surgeries with enhanced outcomes at the University of Missouri. This integration of innovation demonstrates our tireless commitment to ensuring excellence in the comprehensive care of a diverse range of patients with complex spinal pathologies.

A clinical model to predict postoperative improvement in sub-domains of the modified Japanese Orthopedic Association score for degenerative cervical myelopathy.

PURPOSE: The modified Japanese Orthopedic Association (mJOA) score consists of six sub-domains and is used to quantify the severity of cervical myelopathy. The current study aimed to assess for predictors of postoperative mJOA sub-domains scores following elective surgical management for patients with cervical myelopathy and develop the first clinical prediction model for 12-month mJOA sub-domain scores.Please confirm if the author names are presented accurately and in the correct sequence (given name, middle name/initial, family name). Author 1 Given name: [Byron F.] Last name [Stephens], Author 2 Given name: [Lydia J.] Last name [McKeithan], Author 3 Given name: [W. Hunter] Last name [Waddell], Author 4 Given name: [Anthony M.] Last name [Steinle], Author 5 Given name: [Wilson E.] Last name [Vaughan], Author 6 Given name: [Jacquelyn S.] Last name [Pennings], Author 7 Given name: [Jacquelyn S.] Last name [Pennings], Author 8 Given name: [Scott L.] Last name [Zuckerman], Author 9 Given name: [Kristin R.] Last name [Archer], Author 10 Given name: [Amir M.] Last name [Abtahi] Also, kindly confirm the details in the metadata are correct.Last Author listed should be Kristin R. Archer METHODS: A multivariable proportional odds ordinal regression model was developed for patients with cervical myelopathy. The model included patient demographic, clinical, and surgery covariates along with baseline sub-domain scores. The model was internally validated using bootstrap resampling to estimate the likely performance on a new sample of patients. RESULTS: The model identified mJOA baseline sub-domains to be the strongest predictors of 12-month scores, with numbness in legs and ability to walk predicting five of the six mJOA items. Additional covariates predicting three or more items included age, preoperative anxiety/depression, gender, race, employment status, duration of symptoms, smoking status, and radiographic presence of listhesis. Surgical approach, presence of motor deficits, number of surgical levels involved, history of diabetes mellitus, workers' compensation claim, and patient insurance had no impact on 12-month mJOA scores. CONCLUSION: Our study developed and validated a clinical prediction model for improvement in mJOA scores at 12 months following surgery. The results highlight the importance of assessing preoperative numbness, walking ability, modifiable variables of anxiety/depression, and smoking status. This model has the potential to assist surgeons, patients, and families when considering surgery for cervical myelopathy. LEVEL OF EVIDENCE: Level III.

Combined efficacy of Cinnamomum zeylanicum and doxorubicin against leukemia through regulation of TRAIL and NF-kappa B pathways in rat model.

BACKGROUND: Recent discoveries in cancer therapeutics have proven combination therapies more effective than individual drugs. This study describes the efficacy of the combination of Cinnamomum zeylanicum and doxorubicin against benzene-induced leukemia. METHODS AND RESULTS: Brine shrimp assay was used to assess the cytotoxicity of C. zeylanicum, doxorubicin and their combination. After AML induction in Sprague Dawley rats, the same drugs were given to rat groups. Changes in organ weight, haematological profile, and hepatic enzymes were determined. Real-time PCR was used to elucidate the effect on the expression of STMN1, GAPDH, P53 and various TRAIL and NF-kappaB components. C. zeylanicum reduced the cytotoxicity of doxorubicin. The combination treatment showed better anti-leukemic results than any of the individual drugs as evident from STMN1 expression (p < 0.001). It was particularly effective in reducing total white blood cell counts and recovering lymphocytes, monocytes and eosinophils along with hepatic enzymes ALT and AST (p < 0.001). All doses recovered relative organ weights and improved blood parameters. The combination therapy was particularly effective in inducing apoptosis, inhibition of proliferation marker GAPDH (p < 0.001) and NF-kappaB pathway components Rel-A (p < 0.001) and Rel-B (p < 0.01). Expressions of TRAIL components c-FLIP (p < 0.001), TRAIL ligand (p < 0.001) and caspase 8 (p < 0.01) were also altered. CONCLUSION: Cinnamomum zeylanicum in combination with doxorubicin helps to counter benzene-induced cellular and hepatic toxicity and improves haematological profile. The anti-leukemic effects are potentially due to inhibition of GAPDH and NF-kappa B pathway, and through regulation of TRAIL pathway. Our data suggests the use of C. zeylanicum with doxorubicin to improve anti-leukemic therapeutic regimes.

Evaluation of Acacia cyanophylla for their analgesic, anti-pyretic and anti-inflammatory potentials.

The present paper presents results of analgesic, antipyretic activity and anti-inflammatory potential of extract obtained from Acacia cyanophylla when tested at different doses. Analgesic potential of the crude methanolic extract tested by acetic acid assay was dose dependent and maximum activity of 61.60% was measured at 400 mg/kg. Analgesic activity by hot plate method revealed that maximum activity of 36.98% was noted when the mice were exposed to 90 minutes at higher dose of 400 mg/kg. Similar pattern for antipyretic activity was observed as noted for analgesic activity. Anti-inflammatory activity was dose and time dependent when evaluated by Carrageenan-induced paw edema and Xylene-induced ear edema model. Maximum anti-inflammatory activity (43.32%) was shown by crude methanolic extract of Acacia cyanophylla at 400mg/kg-1 after 5 hours on Carrageenan-induced paw edema model. Similarly, maximum (68.80%) anti-inflammatory activity was noted when accessed by Xylene-induced ear edema model at the dose of 200mg/kg after 60 minutes. No in vivo toxicity of the extracts up to the dose of 2000mg/kg was observed using albino mice.

Consanguinity, the driving force behind inheritance of HbS-ß thalassemia in Southern Districts of KP.

OBJECTIVE: To determine the clinical, haematological and genetic factors responsible for variable phenotypes of sickle haemoglobin, sickle haemoglobin-beta, and beta-thalassemia patients. METHODS: The study was conducted in Bannu, Lakki, Tank and Dera Ismail Khan districts of Khyber Pakhtunkhwa province of Pakistan from September 2016 to November 2017, and comprised sickle haemoglobin, sickle haemoglobin-beta, and beta-thalassemia patients. Clinical, haematological and genetic determinants were evaluated using haemoglobin electrophoresis and allele-specific primers through polymerase chain reaction to determine alpha and beta thalassemia, and CgT substitution at position -158 (referred to as Xmn-I polymorphism) in gamma-globin gene. Data was analysed using SPSS 20. RESULTS: Eight b-thalassemia mutations were identified that included IVS I-5(G C), codon 8/9 (+G), codon 30 (G C), -88 (C T), Cap+1(A G), codon 41/42 (-TCTT), IVS I-1(G T) and codon 16(-C). Codon 30 (G C) and -88 (C T) were found only in Pashtoon subjects, Cap+1(A G) and IVS I-1(G T) in Balochi subjects, while 75% of IVS I-5(G C) mutation cases were found in Punjabi ethnic group. In the Pashtoon group, 13 sickle haemoglobin homozygous patients were identified for the first time. Both alpha thalassemia and Xmn-I polymorphism in homozygous condition were common among those with mild phenotype. CONCLUSION: Phenotypic expression of sickle haemoglobin beta thalassemia was found to be extremely variable and alpha thalassemia and Xmn-I polymorphism in homozygous condition were found to be additional genetic modifiers of the disease.

Comparison of 3 Days Amoxicillin Versus 5 Days Co-Trimoxazole for Treatment of Fast-breathing Pneumonia by Community Health Workers in Children Aged 2-59 Months in Pakistan: A Cluster-randomized Trial.

BACKGROUND: Globally, most deaths due to childhood pneumonia occur at the community level. Some countries are still using oral co-trimoxazole, despite a World Health Organization recommendation of oral amoxicillin for the treatment of fast-breathing pneumonia in children at the community level. METHODS: We conducted an unblinded, cluster-randomized, controlled-equivalency trial in Haripur District, Pakistan. Children 2-59 months of age with fast-breathing pneumonia were treated with oral amoxicillin suspension (50 mg/kg/day) for 3 days in 14 intervention clusters and oral co-trimoxazole suspension (8 mg trimethoprim/kg and 40 mg sulfamethoxazole/kg/day) for 5 days in 14 control clusters by lady health workers (LHW). The primary outcome was treatment failure by day 4 for intervention clusters and by day 6 for control clusters. The analysis was per protocol. RESULTS: Out of the 15 749 cases enrolled in the study, 9153 cases in intervention and 6509 cases in control clusters were included in the analysis. Treatment failure rates were 3.6% (326) in intervention clusters and 9.1% (592) in control clusters. After adjusting for clustering, the risk of treatment failure was lower in intervention clusters (risk difference [RD] -5.5%, 95% confidence interval [CI] -7.4--3.7%) than in control clusters. Children with incomplete adherence had a small increase in treatment failure versus those with complete adherence (RD 2.9%, 95% CI 1.6-4.1%). No deaths or serious adverse events occurred. CONCLUSIONS: A 3-day course of oral amoxicillin, administered by LHWs, is an effective and safe treatment for fast-breathing pneumonia in children 2-59 months of age. A shorter course of amoxicillin improves adherence to therapy, is low in cost, and puts less pressure on antimicrobial resistance. CLINICAL TRIALS REGISTRATION: ISRCTN10618300.

Prediction of Oswestry Disability Index (ODI) using PROMIS-29 in a national sample of lumbar spine surgery patients.

PURPOSE: The primary purpose was to examine the measurement properties of the PROMIS-29 to better understand its use in patients undergoing spine surgery. A secondary objective was to calculate a predictive equation between PROMIS-29 and ODI, to allow clinicians and researchers to determine a predicted ODI score based on PROMIS short form scores. METHODS: 719 patients with PROMIS v2.0 and ODI responses were queried from the quality outcomes database. Validity was assessed using coefficient omega, ceiling/floor effects, and confirmatory factor analysis. Multivariable regression predicting ODI scores from PROMIS-29 domains was used to create a predictive equation. Predicted ODI scores were plotted against ODI scores to determine how well PROMIS-29 domains predicted ODI. RESULTS: Results showed good reliability and validity of PROMIS-29 in patients undergoing lumbar spine surgery: convergent and discriminant validity, low floor/ceiling effects, and unidimensional domains. The conversion equation used 6 PROMIS-29 domains (ODI% =  37.847- 1.475*[PFraw] + 1.842*[PAINraw] + 0.557*[SDraw] - 0.642*[SRraw] + 0.478*[PIraw] + 0.295*[DEPraw]). Correlation between the predicted and actual ODI scores was R = 0.88, R2 = 0.78, suggesting that the equation predicted ODI scores that are strongly correlated with actual ODI scores. CONCLUSIONS: Good measurement properties support the use of PROMIS-29 in spine surgery patients. Findings suggest accurate ODI scores can be derived from PROMIS-29 domains. Clinicians who want to move from ODI to PROMIS-29 can use this equation to obtain estimated ODI scores when only collecting PROMIS-29. These results support the idea that PROMIS-29 domains have the potential to replace disease-specific traditional PROMs.

Epidemiological trends and risk factors associated with dengue disease in Pakistan (1980-2014): a systematic literature search and analysis.

BACKGROUND: Dengue is becoming more common in Pakistan with its alarming spreading rate. A historical review needs to be carried out to find the root causes of dengue dynamics, the factors responsible for its spread and lastly to formulate future strategies for its control. METHODS: We searched (January, 2015) all the published literature between 1980 and 2014 to determine spread/burden of dengue disease in Pakistan. RESULTS: A total of 81 reports were identified, showing high numbers of dengue cases in 2010, 2011, and 2013. The tendency of dengue to occur in younger than in older age groups was evident throughout the survey period and all four serotypes were recorded, with DENV1 the least common. Most dengue hemorrhagic fever (DHF) cases fell in the 20-45 years age range. High frequencies tended to be observed first in the Southern coastal region characterized by mild winters and humid warm summers and then the disease progressed towards the lowland areas of the Indus plain with cool winters, hot summers and monsoon rainfall. Based on this survey, new risk maps and infection estimates were identified reflecting public health burden imposed by dengue at the national level. CONCLUSIONS: Our study showed that dengue is common in the three provinces of Pakistan, i.e., Khyber Pakhtunkhwa (KP), Punjab and Sindh. Based on the literature review as well as on our study analysis the current expansion of dengue seems multifactorial and may include climate change, virus evolution, and societal factors such as rapid urbanization, population growth and development, socioeconomic factors, as well as global travel and trade. Due to inadequate remedial strategies, effective vector control measures are essential to target the dengue vector mosquito where high levels of human-vector contact occur. The known social, economic, and disease burden of dengue is alarming globally and it is evident that the wider impact of this disease is grossly underestimated. An international multi-sectoral response, outlined in the WHO Global Strategy for Dengue Prevention and Control, 2012-2020, is now essential to reduce the significant influence of this disease in Dengue endemic areas. Overall gaps were identified in knowledge around seroprevalence, dengue incidence, vector control, genotype evolution and age-stratified serotype circulation.

High-intensity focused ultrasound: past, present, and future in neurosurgery.

Since Lynn and colleagues first described the use of focused ultrasound (FUS) waves for intracranial ablation in 1942, many strides have been made toward the treatment of several brain pathologies using this novel technology. In the modern era of minimal invasiveness, high-intensity focused ultrasound (HIFU) promises therapeutic utility for multiple neurosurgical applications, including treatment of tumors, stroke, epilepsy, and functional disorders. Although the use of HIFU as a potential therapeutic modality in the brain has been under study for several decades, relatively few neuroscientists, neurologists, or even neurosurgeons are familiar with it. In this extensive review, the authors intend to shed light on the current use of HIFU in different neurosurgical avenues and its mechanism of action, as well as provide an update on the outcome of various trials and advances expected from various preclinical studies in the near future. Although the initial technical challenges have been overcome and the technology has been improved, only very few clinical trials have thus far been carried out. The number of clinical trials related to neurological disorders is expected to increase in the coming years, as this novel therapeutic device appears to have a substantial expansive potential. There is great opportunity to expand the use of HIFU across various medical and surgical disciplines for the treatment of different pathologies. As this technology gains recognition, it will open the door for further research opportunities and innovation.

A predictive model and nomogram for predicting return to work at 3 months after cervical spine surgery: an analysis from the Quality Outcomes Database.

OBJECTIVEBack pain and neck pain are two of the most common causes of work loss due to disability, which poses an economic burden on society. Due to recent changes in healthcare policies, patient-centered outcomes including return to work have been increasingly prioritized by physicians and hospitals to optimize healthcare delivery. In this study, the authors used a national spine registry to identify clinical factors associated with return to work at 3 months among patients undergoing a cervical spine surgery.METHODSThe authors queried the Quality Outcomes Database registry for information collected from April 2013 through March 2017 for preoperatively employed patients undergoing cervical spine surgery for degenerative spine disease. Covariates included demographic, clinical, and operative variables, and baseline patient-reported outcomes. Multiple imputations were used for missing values and multivariable logistic regression analysis was used to identify factors associated with higher odds of returning to work. Bootstrap resampling (200 iterations) was used to assess the validity of the model. A nomogram was constructed using the results of the multivariable model.RESULTSA total of 4689 patients were analyzed, of whom 82.2% (n = 3854) returned to work at 3 months postoperatively. Among previously employed and working patients, 89.3% (n = 3443) returned to work compared to 52.3% (n = 411) among those who were employed but not working (e.g., were on a leave) at the time of surgery (p < 0.001). On multivariable logistic regression the authors found that patients who were less likely to return to work were older (age > 56-65 years: OR 0.69, 95% CI 0.57-0.85, p < 0.001; age > 65 years: OR 0.65, 95% CI 0.43-0.97, p = 0.02); were employed but not working (OR 0.24, 95% CI 0.20-0.29, p < 0.001); were employed part time (OR 0.56, 95% CI 0.42-0.76, p < 0.001); had a heavy-intensity (OR 0.42, 95% CI 0.32-0.54, p < 0.001) or medium-intensity (OR 0.59, 95% CI 0.46-0.76, p < 0.001) occupation compared to a sedentary occupation type; had workers' compensation (OR 0.38, 95% CI 0.28-0.53, p < 0.001); had a higher Neck Disability Index score at baseline (OR 0.60, 95% CI 0.51-0.70, p = 0.017); were more likely to present with myelopathy (OR 0.52, 95% CI 0.42-0.63, p < 0.001); and had more levels fused (3-5 levels: OR 0.46, 95% CI 0.35-0.61, p < 0.001). Using the multivariable analysis, the authors then constructed a nomogram to predict return to work, which was found to have an area under the curve of 0.812 and good validity.CONCLUSIONSReturn to work is a crucial outcome that is being increasingly prioritized for employed patients undergoing spine surgery. The results from this study could help surgeons identify at-risk patients so that preoperative expectations could be discussed more comprehensively.

Role of Intra-operative MRI (iMRI) in Improving Extent of Resection and Survival in Patients with Glioblastoma Multiforme.

Multiple intraoperative aids have been introduced to improve the extent of resection (EOR) in Glioblastoma Multiforme (GBM) patients, avoiding any new neurological deficits. Intraoperative MRI (iMRI) has been debated for its utility and cost for nearly two decades in neurosurgical literature. Review of literature suggests improved EOR in GBM patients who underwent iMRI assisted surgical resections leading to higher overall survival (OS) and progression free survival (PFS). iMRI provides real time intraoperative imaging with reasonable quality. Higher risk for new postoperative deficits with increased EOR is not reported in any study using iMRI. The level of evidence regarding prognostic benefits of iMRI is still of low quality..

Prognostic significance of IDH 1 mutation in patients with glioblastoma multiforme.

Focus of brain tumour research is shifting towards tumour genesis and genetics, and possible development of individualized treatment plans. Genetic analysis shows recurrent mutation in isocitrate dehydrogenase (IDH1) gene in most Glioblastoma multiforme (GBM) cells. In this review we evaluated the prognostic significance of IDH 1 mutation on the basis of published evidence. Multiple retrospective clinical analyses correlate the presence of IDH1 mutation in GBM with good prognostic outcomes compared to wild-type IDH1. A systematic review reported similar results. Based on the review of current literature IDH1 mutation is an independent factor for longer overall survival (OS) and progression free survival (PFS) in GBM patients when compared to wild-type IDH1. The prognostic significance opens up new avenues for treatment.

Role of 5-ALA in improving extent of tumour resection in patients with Glioblastoma Multiforme.

Goal of surgery for patients with Glioblastoma Multiforme (GBM) is gross total resection with no new neurological deficits. Surgical resection is often restricted due the difficulty in differentiating the tumour from surrounding normal brain using either naked eye, or standard intra-operative white light microscopy. GBM uptakes orally administered 5-ALA becomes fluorescent when viewed by a special light, and this property has been used to improve intra-operative tumour identification. This technique should therefore allow better extent of tumour resection. The hypothesis has been tested through several studies and even though most studies are of low quality, they strongly favour the use of 5- ALA in improving the extent of resection when compared to white light microscopy. A systematic review on the topic had a similar conclusion. Few studies have also hinted on a high false negative rate with the use of this technique..

Comparison between endoscopic and microscopic approaches for surgery of pituitary tumours.

Surgical techniques for resection of pituitary tumours have come a long way since it was first introduced in late 18th century. Nowadays, most pituitary surgeries are performed through trans-nasal trans-sphenoidal approach either using a microscope, or an endoscope. Herein the authors review the literature and compare these two instruments with regards to their outcomes when used for resection of pituitary tumours. .

Single primer-mediated circular polymerase chain reaction for hairpin DNA cloning and plasmid editing.

We developed and validated a universal polymerase chain reaction (PCR) method, single primer circular (SPC)-PCR, using single primer to simultaneously insert and amplify a short hairpin sequence into a vector with a high success rate. In this method, the hairpin structure is divided into two parts and fused into a vector by PCR. Then, a single primer is used to cyclize the chimera into a mature short hairpin RNA (shRNA) expression vector. It is not biased by loop length or palindromic structures. Six hairpin DNAs with short 4-nucleotide loops were successfully cloned. Moreover, SPC-PCR was also applied to plasmid editing within 3 h with a success rate higher than 95%.

Brevibacillus sediminis sp. nov., isolated from a hot spring.

Strain YIM 78300T, a novel Gram-stain-positive, moderately thermophilic, endospore-forming, rod-shaped, motile bacterium, was recovered from the sediment of a hot spring in the Tagejia Geothermal Field, Angren, Tibet province, western China. Optimum growth was observed at 50-55 °C, at pH 7.0 and with 0-1.5 % (w/v) NaCl. Phylogenetic analysis of the 16S rRNA gene sequence of strain YIM 78300T indicated that it belongs to the genus Brevibacillus. Similarity levels between the 16S rRNA gene sequences of the new isolate and those of the type strains of Brevibacillus members were 96.9-96.3 %; highest sequence similarity was with Brevibacillus thermoruber DSM 7064T. The predominant menaquinone was MK-7 and the major cellular fatty acids were iso-C15 : 0 and iso-C17 : 0. The major polar lipids were phosphatidyl-N-methylethanolamine, phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, two unidentified phospholipids, an unidentified aminophospholipid and two unidentified polar lipids. The G+C content of the genomic DNA of strain YIM 78300T was 57.9 mol%. Based on phylogenetic analyses, and physiological and biochemical characteristics, strain YIM 78300T is considered to represent a novel species of the genus Brevibacillus, for which the name Brevibacillus sediminis sp. nov. is proposed. The type strain is YIM 78300T ( = DSM 29928T = CPCC 100738T).

Abyssicoccus albus gen. nov., sp. nov., a novel member of the family Staphylococcaceae isolated from marine sediment of the Indian Ocean.

A Gram-stain positive, aerobic, non-motile, asporogenous, coccoid shaped bacterium, designated YIM M12140(T), was isolated from a marine sediment sample collected from the Indian Ocean. Phylogenetic analysis showed that strain YIM M12140(T) forms a separate clade within the family Staphylococcaceae. Strain YIM M12140(T) shares high 16S rRNA gene sequence similarity with Macrococcus brunensis DSM 19358(T) (92.9 %). The isolate was found to grow at 0-10 % (w/v) NaCl (optimum, 2-3 %), pH 6.0-10.0 (optimum, pH 8.0) and temperature 5-40 °C (optimum, 28 °C). The polar lipids were identified as diphosphatidylglycerol, phosphatidylglycerol, one unidentified aminophospholipid and two unidentified polar lipids. The major cellular fatty acids of the strain were identified as anteiso-C15:0, -C17:0, iso-C16:0, anteiso-C19:0 and C20:0. The respiratory menaquinones were found to be MK-6 (94 %) and MK-7 (6 %). The cell wall amino acids were found to contain Lys, Ala, Glu, Gly, Asp, Ser and Thr. Whole cell sugars were identified as mannose, ribose, rhamnose, glucose, galactose and xylose. The G+C content of the genomic DNA of strain YIM M12140(T) was determined to be 42.4 mol %. Based on phenotypic, chemotaxonomic data and phylogenetic analysis, it is proposed that strain YIM M12140(T) represents a novel species of a new genus in the family Staphylococcaceae, for which the name Abyssicoccus albus gen. nov., sp. nov. is proposed. The type strain is YIM M12140(T) (= DSM 29158(T) = CCTCC AB 2014213(T)).

Posterior Transdural Approach for Treatment of Spontaneous Intradural Cervical Disc Herniation-Causing Brown-Séquard Syndrome: A 2-Dimensional Operative Video.

Intradural disc herniations (IDH) account for 0.27% of all disc herniations. Cervical IDH account for 3% of these, with only 47 cases reported in the literature, making it an extremely rare diagnosis. Brown-Séquard syndrome is the most common presentation of cervical IDH. Emergent decompression is usually necessary. We present the case of a 56-year-old woman who presented with Brown-Séquard syndrome secondary to a spontaneous intradural C5-6 disc herniation. A posterior transdural approach was recommended to arrest her neurological deficit and promote improvement. The patient consented to the procedure. Institutional Review Board approval was not necessary, given this treatment was necessary and indicated. A standard posterior cervical exposure and C5-6 laminectomies were performed. The dura was opened with a "peeling" technique. The compressive disc fragments were removed. A dural defect was identified along the C6 nerve root sleeve. Fluoroscopy was used to confirm the communication of the defect with the C5-6 disc space. A single 5-0 Prolene suture was used to repair the defect, approximating the anterior dura with a flap from the nerve root sleeve. The patient had an uneventful postoperative course. At 7-month follow-up, her neurological deficits had nearly resolved. Surgeons should consider IDH in the differential diagnosis for sudden neurologic decline in the setting of an intradural mass and should be familiar with the association between cervical IDH and Brown-Séquard syndrome. The posterior transdural approach provides excellent exposure, easier management of dural defects, and ability to handle a variety of intradural pathologies and may avoid the need for concomitant fusion.