Structural Modeling and Validation of Growth/Differentiation Factor 15 [NP_004855] Associated with Pregnancy Complication- Hyperemesis Gravidarum.

Background: Hyperemesis Gravidarum (HG) is acommon pregnancy complication that occurs in0.3–2% of pregnancies. Growth/Differentiation Factor(GDF) 15 serum levels are abnormally high in patientsassociated with HG. In silico analysis providesinformation about structure and function of GDF15.Aim and Objectives: The aim of this study was to enlistbiochemical and functional properties of GDF15protein and determine its three-dimensional structure,as GDF15 is known to be associated with risk of HG.Material and Methods: The PDB file of GDF15[NP_004855] was created by RaptorX structureprediction server. The UCLA-DOE server was used tovisual analysis of crystal structure of protein. Thevalidation for structure models was performed by usingPROCHECK. Model quality estimates were based onthe QMEAN and ProSA. Results: The model showedgood stereo-chemical property in terms of G-factorvalue -0.64, that indicates geometry of modelcorresponds to probability conformation with 95%residue in the favored region of Ramachandran plot,showing high accuracy of model prediction. The Zscore of -4.04 predicted by ProSA represents the goodquality of the model. The energy plot shows the localmodel quality by plotting knowledge-based energies asa function of amino acid sequence position.Conclusion: The generated model could be supportiveto understand the structure and functionalcharacteristics of Homo sapiens growth/differentiationfactor 15 [NP_004855]. As abnormal high serum levelsof GDF15 were observe in patients associated with HG.Therefore, the structure model of GDF15 [NP_004855]is useful to understand its role in development of HG. InSilico docking study could be explain the molecularassociation of GDF15 [NP_004855] with HG and newdrug designing, for that structure model is very useful.

Study of Neuromuscular Transmission Under (i) Phenomenon of Fatigue, (ii) Site of Fatigue, (iii) Neuromuscular Blocking in an in-situ Rat Nerve Muscle Preparation: ANovel Approach to Nerve Muscle Physiology Experiment Teaching

Purpose of the study: Nerve-muscle physiology is a very basic and vital module in undergraduate physiologycurriculum. The practical sessions on this system are demonstrated on amphibian (frog) nerve-musclepreparation, which becomes a limiting factor in most of the medical colleges of India. In this study, wepropose an alternative approach by using mammalian (rat) model for nerve-muscle physiology undergraduatepractical experiments.Materials and methods: Rat in-situ sciatic nerve-soleus muscle preparation was used to study neuromusculartransmission. Stimulation of nerve and recording of muscle contraction (force) were done by using digitalrecording system. To demonstrate fatigue, repetitive electrical stimulation was applied to nerve-musclepreparation and muscle twitches were recorded. A reduction in amplitude of contraction to 50% of their basalrecording was considered as onset of fatigue. To demonstrate site of fatigue, the muscle was stimulateddirectly after the fatigue was observed with nerve stimulation and muscle twitch recorded. To observe theeffect of neuromuscular blocking drug Pancuronium bromide, on neuromuscular transmission, in a separateset up the drug was injected in muscle belly at multiple sites and nerve was stimulated to elicit muscletwitch. The response (amplitude of muscle twitch) was compared with control (injection of 0.9% saline).Main findings: On repeated stimulation of nerve muscle preparation, initially there was an increase inamplitude of contraction but progressively amplitude went on decreasing. After development of fatigue ondirect muscle stimulation, amplitude recorded was same as the initial twitch amplitude. This demonstratesthat the site of fatigue is not the muscle. Further, as nerve is also non- fatigable, site of fatigue wasneuromuscular junction.Injection of Pancuronium bromide showed that the twitch amplitude decreased substantially than the control(saline injection) on stimulation of nerve, but on direct muscle stimulation amplitude of contraction remainedsame as initial, confirming the effect of drug on neuromuscular junction.

Demonstration of Nerve Muscle Preparation in Rats: For Nerve-Muscle Physiology Teaching

Purpose of the study: Physiology practical complement learning of concepts taught in didactic theorylectures. Therefore, they are instrumental to comprehensive learning. Learning of physiology of excitabletissues is supplemented by observing the same phenomena in action in the laboratory. We developed anin-situ rat nerve-muscle model to demonstrate nerve-muscle physiology phenomenon to undergraduate andpostgraduate students. This mimics the physiological conditions unlike the Rana tigrina experiments whichwere done earlier. Further, frequent unavailability of frogs envisaged us to find an alternative for demonstrationof these concepts.Materials and methods: We developed and standardized an in-situ rat soleus muscle-sciatic nerve modelto demonstrate the phenomenon of nerve-muscle properties like simple muscle twitch, effect of strength ofstimulus, effect of two successive stimuli and tetanus to the undergraduate and postgraduate students atour institute.Results: Simple muscle twitch was recorded and effect of increasing strength of stimuli was observed.Thereafter, effect of two successive stimuli and genesis of tetanus was recorded in the same experimentalset up. We successfully standardised this in-situ model for teaching nerve and muscle physiology to our students.Conclusion: We propose an easy and effective experimental rat in-situ model for nerve-muscle physiologyteaching to students

Correlation of High Sensitive C-reactive Protein levels with Mean Arterial Pressure and Serum Calcium in Women with Mild and Severe Pre-eclampsia

Aim:To compare the mean results of serum levels of hs-CRP and calcium in mild and severe pre-eclamptic women with healthy pregnant women and to investigate the relationship, if any between levels of serum hs-CRP with mean arterial pressure (MAP) and serum calcium. Material and Methods:A case control study was performed on 50-50 primigravidas with mild and severe PE as cases as per clinical guidelines and 50 healthy pregnant women as controls. They were all age and parity matched primigravidas at the third trimester of pregnancy. Results:By using ANOVA, statistically highly significant mean values of hs-CRP and serum calcium were observed among all groups (healthy pregnant women, mild pre-eclamptic and severe pre-eclamptic women). Significant positive correlation is found between hs-CRP with MAP, while the negative association is detected among hs-CRP and serum calcium.Conclusion:Serum hs-CRP may be feasible to be used as a sensitive biomarker for determining women at risk of PE. There is a significant influence of calcium supplementation during the antenatal period and the occurrence of pregnancy induced hypertension.

Rat bone marrow stromal cell transplantation ameliorates complete spinal cord injury induced sensorimotor dysfunctions and associated neurotransmitters

Traumatic spinal cord injury (SCI) leads to sensorimotor dysfunction with significant impact on the patient and their family’s quality of life, social, and economic status. There is no complete restorative treatment so far. Bone marrow stromal cells (BMSCs) have anti-inflammatory and neuroprotective effects and recently emerged as a therapeutic candidate for SCI repair. Here, we examined the role of rat BMSCs transplantation on thoracic (T11) complete SCI induced dysfunctions, namely hyperalgesia, allodynia, locomotion, spinal reflexes, and spinal neurotransmitters in rats. Pre-labelled BMSCs were injected on day 9 after SCI locally. We observed that BMSCs transplantation facilitate locomotor recovery (week 2-8) and attenuated hyperalgesia and allodynia to varying sensory stimuli (week 6-8) after SCI. In addition, spinal reflexes and neurotransmitters were affected significantly by complete SCI, which were partially restored by BMSCs transplantation. Histological analyses also revealed the presence of BMSCs at the injury site and appear to fill the lesion cavities, thereby significantly reducing the lesion volume. Our data shows the beneficial effects of BMSCs transplantation on complete SCI-induced sensorimotor functional deficits in rats.

Association of BCL11A genetic variant (rs11886868) with severity in β-thalassaemia major & sickle cell anaemia.

Background & objectives: The amount of foetal haemoglobin that persists in adulthood affects the clinical severity of haemoglobinopathies including β-thalassaemia major and sickle cell anaemia (SCA). The present study was undertaken to analyse β-thalassaemia as well as SCA patients for the single nucleotide polymorphism (SNP), rs11886868 (T/C) in BCL11A gene and to evaluate the association between this polymorphism and severity of β-thalassaemia major and SCA. Methods: A total of 620 samples (420 β-thalassaemia major and 200 SCA cases) were analysed before blood transfusion using basic screening tests like complete blood analysis and osmotic fragility and further confirmed by high performance liquid chromatography (HPLC), amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) and reverse dot blot techniques. All patients were transfusion dependent. Patients with β-thalassaemia and SCA were classified into mild, moderate, severe according to the severity score based on Hb levels, age of onset, age at which patients received their first blood transfusion, the degree of growth retardation and splenectomy. β-thalassaemia as well as SCA patients were analysed for the SNP, rs11886868 (T/C) in BCL11A gene and association between this polymorphism and severity of β-thalassaemia major as well as SCA was evaluated. Results: There was a significant difference in genotypic and allelic frequencies of BCL11A gene polymorphism between mild and moderate and mild and severe cases in both the groups. A significant (P<0.001) difference was observed in the mean HbF levels between the three genotypes in different severity groups. HbF levels were found to be high in CC genotype bearing individuals followed by TC and TT in β-thalassaemia major as well as SCA. Interpretation & conclusions: This study confirms that the T/C variant (rs11886868) of the BCL11A gene causing downregulation of BCL11A gene expression in adult erythroid precursors results in the induction of HbF and ameliorates the severity of β-thalassaemia as well as SCA.

Prenatal music stimulation facilitates the postnatal functional development of the auditory as well as visual system in chicks (Gallus domesticus).

Rhythmic sound or music is known to improve cognition in animals and humans. We wanted to evaluate the effects of prenatal repetitive music stimulation on the remodelling of the auditory cortex and visual Wulst in chicks. Fertilized eggs (0 day) of white leghorn chicken (Gallus domesticus) during incubation were exposed either to music or no sound from embryonic day 10 until hatching. Auditory and visual perceptual learning and synaptic plasticity, as evident by synaptophysin and PSD-95 expression, were done at posthatch days (PH) 1, 2 and 3. The number of responders was significantly higher in the music stimulated group as compared to controls at PH1 in both auditory and visual preference tests. The stimulated chicks took significantly lesser time to enter and spent more time in the maternal area in both preference tests. A significantly higher expression of synaptophysin and PSD-95 was observed in the stimulated group in comparison to control at PH1-3 both in the auditory cortex and visual Wulst. A significant inter-hemispheric and gender-based difference in expression was also found in all groups. These results suggest facilitation of postnatal perceptual behaviour and synaptic plasticity in both auditory and visual systems following prenatal stimulation with complex rhythmic music.

Role of sound stimulation in reprogramming brain connectivity.

Sensory stimulation has a critical role to play in the development of an individual. Environmental factors tend to modify the inputs received by the sensory pathway. The developing brain is most vulnerable to these alterations and interacts with the environment to modify its neural circuitry. In addition to other sensory stimuli, auditory stimulation can also act as external stimuli to provide enrichment during the perinatal period. There is evidence that suggests that enriched environment in the form of auditory stimulation can play a substantial role in modulating plasticity during the prenatal period. This review focuses on the emerging role of prenatal auditory stimulation in the development of higher brain functions such as learning and memory in birds and mammals. The molecular mechanisms of various changes in the hippocampus following sound stimulation to effect neurogenesis, learning and memory are described. Sound stimulation can also modify neural connectivity in the early postnatal life to enhance higher cognitive function or even repair the secondary damages in various neurological and psychiatric disorders. Thus, it becomes imperative to examine in detail the possible ameliorating effects of prenatal sound stimulation in existing animal models of various psychiatric disorders, such as autism.

Effect of magnetic field on food and water intake and body weight of spinal cord injured rats.

Chronic (2h/d × 8 weeks) exposure to magnetic field (MF; 50 Hz, 17.9 T) in complete spinal cord (T13) transected rats restored food intake (FI), water intake (WI) and body weight (BW) which were decreased in the spinal cord injured rats. The results suggest a significant beneficial effect of chronic exposure to magnetic field of paraplegic rats.

Amygdalar neuronal responses to peripheral nociceptive stimuli in rats.

Amygdala plays a very important role in the mediation of pain. In the present study the behaviour of the amygdalar neurons in response to various peripheral noxious stimuli was observed. Noxious mechanical, thermal, electrical, chemical and the non-noxious stimuli (touch) were applied individually to the animal and then the neuronal responses to these stimuli were recorded. Our results showed that the majority of amygdalar units recorded from medial, lateral and basolateral nuclei, responded to different peripheral noxious (thermal, electrical, chemical mechanical) and non-noxious stimuli by excitation. However few neurons decreased their activity on stimulation. Some of these neurons also exhibited after discharge following application of higher intensity of noxious stimuli.

Effect of prenatal species-specific and music stimulation on the postnatal auditory preference of domestic chick.

Perinatal sensory experience plays an important role in the development of perceptual preferences. In the present study prenatal enrichment with sound stimulus was given to see its effect on the development of postnatal auditory preference. Auditory stimulation with either species-specific (chicken maternal and hatching calls) or music (slow and fast sitar music) sounds was provided to two separate sets of fertilized eggs from the day 10 of incubation. The postnatal auditory preference of the chicks to either species-specific or music sounds was then tested at different time periods after hatching. All the chicks, irrespective of the type of prenatal exposure, showed preference for species-specific maternal calls. Notably, the music stimulated chicks did not show preference for either slow or fast music. In both the experimental groups, the number of chicks responding to the species-specific maternal calls was significantly (P<0.001) more at 24 h and 48 h post hatch, when compared with the unstimulated control group. Comparison of the species-specific stimulated group with the music stimulated group, for auditory preference to the maternal calls, did not show any significant difference. Further, in the species-specific sound stimulated groups, there was a significant (P<0.001) increase in the number of chicks responding to maternal calls at 60 h of age with repeated testing. However, there was no effect of peer imprinting on the auditory preference of the chicks, in both the experimental groups. The results indicate that prenatal auditory experience with either species-specific or non-specific music enhances the postnatal auditory preference of chicks for the species-specific sounds.

Dystrophin gene deletions in South Indian Duchenne muscular dystrophy patients.

66 unrelated patients from Southern India with Duchenne Muscular Dystrophy (DMD) were studied for intragenic deletion in 18 exons and Pm region of the DMD gene using multiplex PCR. Of these 41 (62.1%) showed intragenic deletions. 78% of the deletions were located at the distal hotspot region (44-55 exons) and 22% of the deletions were located at the proximal region (exon 2-19). Exon 50 is most frequently deleted. Deletions in isolated cases were significantly more compared to familial cases. The lower incidence reported from South India compared to North India, is suggestive of variations in the Southern and Northern population.

Analgesia in phasic and tonic pain tests in a pharmacological model of autotomy.

Self-mutilation or self-injurious behaviour is a well known behavioural disorder in humans. The proposition that this behaviour in animals is a response to chronic pain of peripheral nerve injury has been met with controversy. In the present study a pharmacological model, which produces no sensory or motor loss was used to study how autotomy is related to pain. In a group of rats autotomy was induced by amphetamine in phenoxybenzamine and reserpine treated animals. The pain tests, both phasic and tonic were then performed. The results of this study showed that a total analgesia was produced in both phasic and tonic pain tests, in animals that exhibited autotomy. Injection of naloxone in these animals prevented autotomy. A correlation between autotomy and no pain is suggested in this pharmacological model of autotomy.