1.An Improved Methodology to Overcome Key Issues in Human Fecal Metagenomic DNA Extraction
Kumar JITENDRA ; Kumar MANOJ ; Gupta SHASHANK ; Ahmed VASIM ; Bhambi MANU ; Pandey RAJESH ; Chauhan Singh NAR
Genomics, Proteomics & Bioinformatics 2016;14(6):371-378
Microbes are ubiquitously distributed in nature, and recent culture-independent studies have highlighted the significance of gut microbiota in human health and disease. Fecal DNA is the primary source for the majority of human gut microbiome studies. However, further improvement is needed to obtain fecal metagenomic DNA with sufficient amount and good quality but low host genomic DNA contamination. In the current study, we demonstrate a quick, robust, unbiased, and cost-effective method for the isolation of high molecular weight (>23 kb) metagenomic DNA (260/280 ratio >1.8) with a good yield (55.8 ± 3.8 ng/mg of feces). We also confirm that there is very low human genomic DNA contamination (eubacterial:human genomic DNA marker genes=227.9:1) in the human feces. The newly-developed method robustly performs for fresh as well as stored fecal samples as demonstrated by 16S rRNA gene sequencing using 454 FLX+. Moreover, 16S rRNA gene analysis indicated that compared to other DNA extraction methods tested, the fecal metagenomic DNA isolated with current methodology retains species richness and does not show microbial diversity biases, which is further confirmed by qPCR with a known quantity of spike-in genomes. Overall, our data highlight a protocol with a balance between quality, amount, user-friendliness, and cost effectiveness for its suitability toward usage for cultureindependent analysis of the human gut microbiome, which provides a robust solution to overcome key issues associated with fecal metagenomic DNA isolation in human gut microbiome studies.
2.Three-dimensional computer navigation in the reconstruction of complex unilateral orbital fractures: evaluation and review of applications
Parampreet Singh SAINI ; Rajesh KUMAR ; Manu SAINI ; Tarush GUPTA ; Sunil GABA ; Ramesh Kumar SHARMA
Archives of Craniofacial Surgery 2024;25(4):161-170
Background:
The eyes are the central aesthetic unit of the face. Maxillofacial trauma can alter facial proportions and affect visual function with varying degrees of severity. Conventional approaches to reconstruction have numerous limitations, making the process challenging. The primary objective of this study was to evaluate the application of three-dimensional (3D) navigation in complex unilateral orbital reconstruction.
Methods:
A prospective cohort study was conducted over 19 months (January 2020 to July 2021), with consecutive enrollment of 12 patients who met the inclusion criteria. Each patient was followed for a minimum period of 6 months. The principal investigator carried out a comparative analysis of several factors, including fracture morphology, orbital volume, globe projection, diplopia, facial morphic changes, lid retraction, and infraorbital nerve hypoesthesia.
Results:
Nine patients had impure orbital fractures, while the remainder had pure fractures. The median orbital volume on the normal side (30.12 cm3; interquartile range [IQR], 28.45–30.64) was comparable to that of the reconstructed orbit (29.67 cm3; IQR, 27.92–31.52). Diplopia improved significantly (T(10) = 2.667, p = 0.02), although there was no statistically significant improvement in globe projection. Gross symmetry of facial landmarks was achieved, with comparable facial width-to-height ratio and palpebral fissure lengths. Two patients reported infraorbital hypoesthesia at presentation, which persisted at the 6-month follow-up. Additionally, five patients developed lower lid retraction (1–2 mm), and one experienced implant impingement at the infraorbital border.
Conclusion
Our study provides level II evidence supporting the use of 3D navigation to improve surgical outcomes in complex orbital reconstruction.
3.Three-dimensional computer navigation in the reconstruction of complex unilateral orbital fractures: evaluation and review of applications
Parampreet Singh SAINI ; Rajesh KUMAR ; Manu SAINI ; Tarush GUPTA ; Sunil GABA ; Ramesh Kumar SHARMA
Archives of Craniofacial Surgery 2024;25(4):161-170
Background:
The eyes are the central aesthetic unit of the face. Maxillofacial trauma can alter facial proportions and affect visual function with varying degrees of severity. Conventional approaches to reconstruction have numerous limitations, making the process challenging. The primary objective of this study was to evaluate the application of three-dimensional (3D) navigation in complex unilateral orbital reconstruction.
Methods:
A prospective cohort study was conducted over 19 months (January 2020 to July 2021), with consecutive enrollment of 12 patients who met the inclusion criteria. Each patient was followed for a minimum period of 6 months. The principal investigator carried out a comparative analysis of several factors, including fracture morphology, orbital volume, globe projection, diplopia, facial morphic changes, lid retraction, and infraorbital nerve hypoesthesia.
Results:
Nine patients had impure orbital fractures, while the remainder had pure fractures. The median orbital volume on the normal side (30.12 cm3; interquartile range [IQR], 28.45–30.64) was comparable to that of the reconstructed orbit (29.67 cm3; IQR, 27.92–31.52). Diplopia improved significantly (T(10) = 2.667, p = 0.02), although there was no statistically significant improvement in globe projection. Gross symmetry of facial landmarks was achieved, with comparable facial width-to-height ratio and palpebral fissure lengths. Two patients reported infraorbital hypoesthesia at presentation, which persisted at the 6-month follow-up. Additionally, five patients developed lower lid retraction (1–2 mm), and one experienced implant impingement at the infraorbital border.
Conclusion
Our study provides level II evidence supporting the use of 3D navigation to improve surgical outcomes in complex orbital reconstruction.
4.Three-dimensional computer navigation in the reconstruction of complex unilateral orbital fractures: evaluation and review of applications
Parampreet Singh SAINI ; Rajesh KUMAR ; Manu SAINI ; Tarush GUPTA ; Sunil GABA ; Ramesh Kumar SHARMA
Archives of Craniofacial Surgery 2024;25(4):161-170
Background:
The eyes are the central aesthetic unit of the face. Maxillofacial trauma can alter facial proportions and affect visual function with varying degrees of severity. Conventional approaches to reconstruction have numerous limitations, making the process challenging. The primary objective of this study was to evaluate the application of three-dimensional (3D) navigation in complex unilateral orbital reconstruction.
Methods:
A prospective cohort study was conducted over 19 months (January 2020 to July 2021), with consecutive enrollment of 12 patients who met the inclusion criteria. Each patient was followed for a minimum period of 6 months. The principal investigator carried out a comparative analysis of several factors, including fracture morphology, orbital volume, globe projection, diplopia, facial morphic changes, lid retraction, and infraorbital nerve hypoesthesia.
Results:
Nine patients had impure orbital fractures, while the remainder had pure fractures. The median orbital volume on the normal side (30.12 cm3; interquartile range [IQR], 28.45–30.64) was comparable to that of the reconstructed orbit (29.67 cm3; IQR, 27.92–31.52). Diplopia improved significantly (T(10) = 2.667, p = 0.02), although there was no statistically significant improvement in globe projection. Gross symmetry of facial landmarks was achieved, with comparable facial width-to-height ratio and palpebral fissure lengths. Two patients reported infraorbital hypoesthesia at presentation, which persisted at the 6-month follow-up. Additionally, five patients developed lower lid retraction (1–2 mm), and one experienced implant impingement at the infraorbital border.
Conclusion
Our study provides level II evidence supporting the use of 3D navigation to improve surgical outcomes in complex orbital reconstruction.
5.Three-dimensional computer navigation in the reconstruction of complex unilateral orbital fractures: evaluation and review of applications
Parampreet Singh SAINI ; Rajesh KUMAR ; Manu SAINI ; Tarush GUPTA ; Sunil GABA ; Ramesh Kumar SHARMA
Archives of Craniofacial Surgery 2024;25(4):161-170
Background:
The eyes are the central aesthetic unit of the face. Maxillofacial trauma can alter facial proportions and affect visual function with varying degrees of severity. Conventional approaches to reconstruction have numerous limitations, making the process challenging. The primary objective of this study was to evaluate the application of three-dimensional (3D) navigation in complex unilateral orbital reconstruction.
Methods:
A prospective cohort study was conducted over 19 months (January 2020 to July 2021), with consecutive enrollment of 12 patients who met the inclusion criteria. Each patient was followed for a minimum period of 6 months. The principal investigator carried out a comparative analysis of several factors, including fracture morphology, orbital volume, globe projection, diplopia, facial morphic changes, lid retraction, and infraorbital nerve hypoesthesia.
Results:
Nine patients had impure orbital fractures, while the remainder had pure fractures. The median orbital volume on the normal side (30.12 cm3; interquartile range [IQR], 28.45–30.64) was comparable to that of the reconstructed orbit (29.67 cm3; IQR, 27.92–31.52). Diplopia improved significantly (T(10) = 2.667, p = 0.02), although there was no statistically significant improvement in globe projection. Gross symmetry of facial landmarks was achieved, with comparable facial width-to-height ratio and palpebral fissure lengths. Two patients reported infraorbital hypoesthesia at presentation, which persisted at the 6-month follow-up. Additionally, five patients developed lower lid retraction (1–2 mm), and one experienced implant impingement at the infraorbital border.
Conclusion
Our study provides level II evidence supporting the use of 3D navigation to improve surgical outcomes in complex orbital reconstruction.
6.Predictive Validity of Some Common Animal Models of Bipolar Disorder Using Lithium and Lamotrigine Therapy: An Attempt towards a Battery-Based Approach for the Evaluation of Mood Stabilizers.
Manu KUMAR ; Chakra Dhar TRIPATHI ; Veena VERMA ; Biswa Mohan PADHY ; Girish Gulab MESHRAM ; B ABHILASH
Psychiatry Investigation 2016;13(4):434-439
OBJECTIVE: To determine the predictive validity of some of the commonly employed models of mania and depression using standard drugs i.e. lithium (70 mg/kg) and lamotrigine (5 mg/kg) in male Wistar rats. METHODS: The depression facet of bipolar disorder was evaluated using forced swim test, tail suspension test, and chronic mild stress test. The models used to evaluate the mania facet of bipolar disorder were isolation-induced aggression test, saccharine preference test, and morphine-sensitized hyperlocomotion test. RESULTS: The immobility time was significantly (p<0.05) reduced by lamotrigine in the tail suspension test and the forced swim test, while lithium caused significant (p<0.05) reduction only in the tail suspension test. Rats exposed to chronic mild stress showed the maximal increment of 1% sucrose consumption at the 3rd week of treatment in both the lithium (p<0.001) and lamotrigine (p<0.01) groups. In the isolation-induced aggression test, the aggressive behaviour of rats was significantly reduced by both lithium [approach (p<0.001), attack (p<0.01), and bite (p<0.01)] and lamotrigine [approach (p<0.001), and attack (p<0.05)]. Neither of the drugs were effective in the saccharine preference test. Only lithium was able to significantly (p<0.05) reduce the crossing parameter in morphine-sensitized rats. CONCLUSION: Our study identifies the chronic mild stress test and isolation-induced aggression test of having the highest predictive validity in the depression and mania facets of bipolar disorder, respectively, and should be a part of a battery of tests used to evaluate novel mood stabilizers.
Aggression
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Animals*
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Bipolar Disorder*
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Depression
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Exercise Test
;
Hindlimb Suspension
;
Humans
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Lithium*
;
Male
;
Models, Animal*
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Rats
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Rats, Wistar
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Saccharin
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Sucrose