Development of EST-SSR markers in Cenchrus ciliaris and their applicability in studying the genetic diversity and cross-species transferability

Most of the grasses of the genus Cenchrus (20–25 species) and Pennisetum (80–140 species) are distributed throughout the tropical and subtropical regions of the world and reproduce both by sexual and apomictic modes. However, the relationships among the Cenchrus–Pennisetum species are not very clear yet. Molecular markers like expressed sequence tag-simple sequence repeats (EST-SSRs) have been reported to be a better choice for resolving the phylogenetic relationships and to estimate the genetic diversity. The present study describes the identification of EST-SSR markers based on the transcriptome data of Cenchrus ciliaris inflorescence and illustrates the genetic diversity and phylogenetic relationships among these species. Of the 378 primer pairs used across 33 accessions of 21 Cenchrus, Pennisetum, and related grass (Bothriochloa, Dichanthium and Panicum) species, 116 EST-SSR markers were found to be polymorphic with an average polymorphism information content (PIC) of 0.49. Fifty-one EST-SSR loci and 520 alleles showed that where the PIC value is >0.5 there the GAG repeat motif was highly polymorphic. Two EST-SSR markers, CcSSR_80 and CcSSR_102, are polymorphic among the Cenchrus species, while they are absent in Pennisetum and the allied species. Five SSR markers (CcSSR_75, CcSSR_85, CcSSR_87, CcSSR_88 and CcSSR_114) showed 100% cross-transferability among the 21 Cenchrus–Pennisetum species. Species-specific alleles could also be detected for seven species of Cenchrus, Pennisetum and Panicum across 10 SSR markers. Assay of polymorphism across these agamic complexes showed that the three SSR markers (CcSSR_26, CcSSR_97 and CcSSR_109) were associated with Cenchrus–Pennisetum complex, and one (CcSSR_47) with Bothriochloa–Dichanthium complex. Markers with high discriminating power, namely CcSSR_4, CcSSR_38, CcSSR_48, CcSSR_66, CcSSR_67 and CcSSR_70, can be used to estimate the allelic sequence divergence across the sexual and apomictic lineages. Genetic diversity analysis using neighbour-joining (NJ) and principal co-ordinate analysis (PCoA) based approaches showed six and five clusters for the 33 accessions, respectively, having congruence in the pattern of clustering. These accessions were grouped according to their mode of reproduction. Cenchrus and Pennisetum species were grouped separately within the same clade, implying monophyletic group within a ‘bristle clade’. Thus, this study showed high discrimination power of microsatellite (EST-SSR) markers to resolve the phylogenetic relationships.

Phase Changing Material for Therapeutic Hypothermia in Neonates with Hypoxic Ischemic Encephalopathy – A Multi-centric Study

Objective: To assess the feasibility and safety of coolingasphyxiated neonates using phase changing material baseddevice across different neonatal intensive care units in India.Design: Multi-centric uncontrolled clinical trial.Setting: 11 level 3 neonatal units in India from November 2014 toDecember 2015.Participants: 103 newborn infants with perinatal asphyxia,satisfying pre-defined criteria for therapeutic hypothermia.Intervention: Therapeutic hypothermia was provided usingphase changing material based device to a target temperature of33.5±0.5oC, with a standard protocol. Core body temperature wasmonitored continuously using a rectal probe during the coolingand rewarming phase and for 12 hours after the rewarming wascomplete.Outcome measures: Feasibility measure - Time taken to reachtarget temperature, fluctuation of the core body temperatureduring the cooling phase and proportion of temperaturerecordings outside the target range. Safety measure - adverseevents during coolingResults: The median (IQR) of time taken to reach targettemperature was 90 (45, 120) minutes. The mean (SD) deviationof temperature during cooling phase was 33.5 (0.39) ºC.Temperature readings were outside the target range in 10.8%(5.1% of the readings were <33oC and 5.7% were >34oC). Mean(SD) of rate of rewarming was 0.28 (0.13)oC per hour. Thecommon adverse events were shock/ hypotension (18%),coagulopathy (21.4%), sepsis/probable sepsis (20.4%) andthrombocytopenia (10.7%). Cooling was discontinued before 72hours in 18 (17.5%) babies due to reasons such as hemodynamicinstability/refractory shock, persistent pulmonary hypertension orbleeding. 7 (6.8%) babies died during hospitalization.Conclusion: Using phase changing material based coolingdevice and a standard protocol, it was feasible and safe to providetherapeutic hypothermia to asphyxiated neonates across differentneonatal units in India. Maintenance of target temperature wascomparable to standard servo-controlled equipment.

Clinical Profile of Swine Flu in Children at Puducherry.

Objective To study the clinical profile and outcome of children screened and diagnosed for Swine flu at a tertiary care hospital. Methods All Children with suspicion of swine flu infection attending our hospital during the epidemic from August 2009 through January 2010 were screened and categorized into A, B and C as per guidelines of National Health and family welfare. Patients in Category A were advised home quarantine, Category B tested and treated with ostelamivir and Category C were hospitalized. Results Among 424 cases screened for Swine flu, 79 were children in the age group 0–12 years of age (18.6%). The median age of presentation was 5 years. Children belonging to Category A were 43(54.4%), Category B were 31 (39.2%) and Category C were 5(6.3%). Out of the cases that were screened, 19 children were confirmed positive for H1N1 infection(30.2%).Out of positive cases 8 were in category A (42%), eight were in category B (42%) and five were in category C (26.3%). Conclusions During the epidemic of swine flu at puducherry, majority of cases were category A with mild symptoms. Home quarantine and preventive measures during the epidemic were found to be far more important than testing and treating with Oseltamivir.

Growth hormone in intra-uterine growth retarded newborns.

OBJECTIVE: To study growth hormone levels in IUGR and healthy controls and its association with birth weight and ponderal index. METHODS: We studied 50 Intra uterine growth retarded (IUGR) and 50 healthy newborns born at term by vaginal delivery in JIPMER, Pondicherry, India. Cord blood was collected at the time of delivery for measurement of growth hormone. RESULTS: When compared with healthy newborns, IUGR newborns had higher growth hormone levels (mean +/- SD, 23.5 +/- 15.6 vs 16.2 +/- 7.61 ngm/ml, P = 0.019). A negative correlation was identified between growth hormone levels and birth weight (r2 = - 0.22, P = 0.03) and ponderal index (r2 = - 0.36, P = 0.008). Correlation of growth hormone levels was much more confident with ponderal index than with birth weight. CONCLUSION: At birth IUGR infants display increased growth hormone levels which correlate with ponderal index much more confidently than with birth weight.

Neurobehavioral assessment of appropriate for gestational and small for gestational age babies.

he present study was conducted to evaluate the neurobehaviour of term appropriate for gestational aland small for gestational age babies during the first two weeks of life in a tertiary care hospital. Forty eight appropriate and thirty small for gestation age babies were evaluated using Brazelton Neurobehavioural Assessment Scale on 3rd, 7th and 14th day of life. The behaviour of AGA babies is characterized by optimal performance in habituation, range of state, regulation of state and autonomic stability. The behavior is at low to mid-range in orientation and in motor clusters. All the behavior clusters showed improvement over first 14 days except for regulation of state which showed a lower performance on day 7 and 14. The behavior performance of SGA babies on day 3, compared to AGA babies, was lower in all the clusters except orientation where they performed much better. The percentage improvement of scores in SGA babies is higher than in AGA babies and by day 14 SGA babies are scoring higher than AGA babies in orientation, autonomic stability and regulation of state. The difference in the neurobehavior pattern of babies in relation to their intrauterine growth suggests need for appropriate care.