A <i>Medicago truncatula</i> Autoregulation of Nodulation Mutant Transcriptome Analysis Reveals Disruption of the SUNN Pathway Causes Constitutive Expression Changes in Some Genes, but Overall Response to Rhizobia Resembles Wild-Type, Including Induction of <i>TML1</i> and <i>TML2</i>

Nodule number regulation in legumes is controlled by a feedback loop that integrates nutrient and rhizobia symbiont status signals to regulate nodule development. Signals from the roots are perceived by shoot receptors, including a CLV1-like receptor-like kinase known as SUNN in Medicago truncatula. In the absence of functional SUNN, the autoregulation feedback loop is disrupted, resulting in hypernodulation. To elucidate early autoregulation mechanisms disrupted in SUNN mutants, we searched for genes with altered expression in the loss-of-function sunn-4 mutant and included the rdn1-2 autoregulation mutant for comparison. We identified constitutively altered expression of small groups of genes in sunn-4 roots and in sunn-4 shoots. All genes with verified roles in nodulation that were induced in wild-type roots during the establishment of nodules were also induced in sunn-4, including autoregulation genes TML2 and TML1. Only an isoflavone-7-O-methyltransferase gene was induced in response to rhizobia in wild-type roots but not induced in sunn-4. In shoot tissues of wild-type, eight rhizobia-responsive genes were identified, including a MYB family transcription factor gene that remained at a baseline level in sunn-4; three genes were induced by rhizobia in shoots of sunn-4 but not wild-type. We cataloged the temporal induction profiles of many small secreted peptide (MtSSP) genes in nodulating root tissues, encompassing members of twenty-four peptide families, including the CLE and IRON MAN families. The discovery that expression of TML2 in roots, a key factor in inhibiting nodulation in response to autoregulation signals, is also triggered in sunn-4 in the section of roots analyzed, suggests that the mechanism of TML regulation of nodulation in M. truncatula may be more complex than published models

Challenges in screening for sickle cell disease among newborns from the tribal region of Palghar, Maharashtra during the COVID-19 pandemic

Background & objectives: Despite several adversities imposed by the COVID-19 pandemic, it was crucial to sustain research having public health relevance such as investigations around sickle cell disease (SCD). Against this background, an ongoing ICMR-multicentric study for newborn screening of SCD in the tribal population at Model Rural Health Research Unit (MRHRU-Dahanu) in Palghar District, Maharashtra constituted the current study setting. This was a descriptive study wherein, certain measures were undertaken and strategies were developed in view of the challenges in newborn screening for SCD due to the COVID-19 pandemic during December 2019-September 2021 at Sub District Hospital, MRHRU-Dahanu. Methods: During the onset of the pandemic, (December 2019-March 2020), the follow up was possible in 26.7 per cent (20/75) of the newborns. Subsequently, challenges such as travel restrictions, fear of COVID-19, shortage of staff were experienced with respect to enrolment and follow up visits. Results: After implementing certain pragmatic strategies (ASHA involvement, usage of virtual platform and flexible visits), follow up rate increased to 47.5 per cent (66/139) between July 2020-April 2021 (post first lockdown) and to 66 per cent (65/98) during the second wave (May 2021-August 2021). Interpretation & conclusions: The study emphasizes the importance of network building, use of virtual platform and engaging health workers in tribal settings. Such pragmatic approaches have the potential to pave a path for further implementation research involving specific interventions to improve health outcomes in tribal settings

Laser Capture Microdissection Transcriptome Reveals Spatiotemporal Tissue Gene Expression Patterns of Medicago truncatula Roots Responding to Rhizobia

We report a public resource for examining the spatiotemporal RNA expression of 54,893 Medicago truncatula genes during the first 72 h of response to rhizobial inoculation. Using a methodology that allows synchronous inoculation and growth of more than 100 plants in a single media container, we harvested the same segment of each root responding to rhizobia in the initial inoculation over a time course, collected individual tissues from these segments with laser capture microdissection, and created and sequenced RNA libraries generated from these tissues. We demonstrate the utility of the resource by examining the expression patterns of a set of genes induced very early in nodule signaling, as well as two gene families (CLE peptides and nodule specific PLAT-domain proteins) and show that despite similar whole-root expression patterns, there are tissue differences in expression between the genes. Using a rhizobial response dataset generated from transcriptomics on intact root segments, we also examined differential temporal expression patterns and determined that, after nodule tissue, the epidermis and cortical cells contained the most temporally patterned genes. We circumscribed gene lists for each time and tissue examined and developed an expression pattern visualization tool. Finally, we explored transcriptomic differences between the inner cortical cells that become nodules and those that do not, confirming that the expression of 1-aminocyclopropane-1-carboxylate synthases distinguishes inner cortical cells that become nodules and provide and describe potential downstream genes involved in early nodule cell division. [Graphic: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license

Predictors of bisexual behaviour among MSM attending intervention sites may help in prevention interventions for this bridge to the heterosexual epidemic in India: data from HIV sentinel surveillance.

Indian cultural tradition demanding marriage, many MSM howsoever they self-identify are likely to be married or have sex with women. To consolidate India's HIV prevention gains, it is important to understand and address the interaction between the MSM and heterosexual epidemics in India and create specific interventions for bisexual MSM. The challenge is to identify and intervene this hard to reach population. Data from HIV Sentinel Surveillance 2011 among MSM in four Indian states were analyzed to assess predictors and prevalence of bisexual behaviour in MSM.Between March-May 2011, 4682 men (15-49 years) who had anal/oral sex with a male partner in the past month, attending intervention sites and consenting for an un-linked anonymous survey answered an 11- item questionnaire and provided blood for HIV test by finger stick at 19 designated surveillance sites.Of 4682 MSM tested overall, 5% were illiterate, 51% reported only receptive anal intercourse, 21% only penetrative and 28% both. 36% MSM had ever received money for sex. Overall 6.8% were HIV infected. 44% MSM were bisexual in the last six months. On multivariate analysis, 'being bisexual' was found to be independently associated with 'older age': 26-30 years [AOR = 3.1, 95% CI(2.7, 3.7)], >30 years [AOR = 6.5, 95% CI(5.5, 7.7)]; 'reporting penetrative behaviour alone' with other men [AOR = 5.8, 95% CI(4.8, 7.0), p<0.01] and 'reporting both penetrative and receptive behaviour' [AOR = 2.7, 95% CI(2.3, 3.1) p<0.01]. Those who both paid and received money for sex [AOR = 0.49, 95% CI (0.38, 0.62)] were significantly less likely to be bisexual.A substantial proportion of men receiving services from Targeted Intervention programs are bisexual and the easy opportunity for intervention in this setting should be capitalised upon. Focusing on older MSM, as well as MSM who show penetrative behaviour with other men, could help in reaching this population

Characterization of the recombination activities of the \u3ci\u3eEntamoeba histolytica\u3c/i\u3e Rad51 recombinase

The protozoan parasite responsible for human amoebiasis is Entamoeba histolytica. An important facet of the life cycle of E. histolytica involves the conversion of the mature trophozoite to a cyst. This transition is thought to involve homologous recombination (HR), which is dependent upon the Rad51 recombinase. Here, a biochemical characterization of highly purified ehRad51 protein is presented. The ehRad51 protein preferentially binds ssDNA, forms a presynaptic filament and possesses ATP hydrolysis activity that is stimulated by the presence of DNA. Evidence is provided that ehRad51 catalyzes robust DNA strand exchange over at least 5.4 kilobase pairs. Although the homologous DNA pairing activity of ehRad51 is weak, it is strongly enhanced by the presence of two HR accessory cofactors, calcium and Hop2-Mnd1. The biochemical system described herein was used to demonstrate the potential for targeting ehRad51 with two small molecule inhibitors of human RAD51. We show that 4,4′-diisothiocyanostilbene-2,2′-disulfonic acid (DIDS) inhibited ehRad51 by interfering with DNA binding and attenuated encystation in Entamoeba invadens, while B02 had no effect on ehRad51 strand exchange activity. These results provide insight into the underlying mechanism of homology-directed DNA repair in E. histolytica

Demographic and socio-economic characteristics of MSM in states in west-central India (Year 2011).

<p>Note: MSMO: Men having Sex with Men Only, MSMW: Men having Sex with Men and Women, IDU: Injecting Drug Use, *Approx. 0.3%–1.3% data on these variables is missing overall.</p><p>Demographic and socio-economic characteristics of MSM in states in west-central India (Year 2011).</p

Predictors of HIV infection among MSM in west-central India (Year 2011).

<p>Note: Column 2: Denominators for percentages are respective figures in 1st column. OR: Odds Ratio; AOR: Adjusted Odds Ratio; IDU: Injecting Drug Use. * ‘Received Money’ = Only sold sex, ‘Paid’ = Only bought sex; ‘Received+Paid’ =  Bought and sold sex.</p><p>Predictors of HIV infection among MSM in west-central India (Year 2011).</p

Predictors of bisexual behaviour among MSM in west-central India (Year 2011).

<p>Note: Column 2: Denominators for percentages are respective figures in 1st column. OR: Odds Ratio; AOR: Adjusted Odds Ratio. * Missing information about bisexuality for 32 records; IDU: Injecting Drug Use, MSMW: Men having Sex with Men and Women. * ‘Received Money’ =  Only sold sex, ‘Paid’ = Only bought sex; ‘Received+Paid’ =  Bought and sold sex.</p><p>Predictors of bisexual behaviour among MSM in west-central India (Year 2011).</p

Characterization of the recombination activities of the Entamoeba histolytica Rad51 recombinase

The protozoan parasite responsible for human amoebiasis is Entamoeba histolytica. An important facet of the life cycle of E. histolytica involves the conversion of the mature trophozoite to a cyst. This transition is thought to involve homologous recombination (HR), which is dependent upon the Rad51 recombinase. Here, a biochemical characterization of highly purified ehRad51 protein is presented. The ehRad51 protein preferentially binds ssDNA, forms a presynaptic filament and possesses ATP hydrolysis activity that is stimulated by the presence of DNA. Evidence is provided that ehRad51 catalyzes robust DNA strand exchange over at least 5.4 kilobase pairs. Although the homologous DNA pairing activity of ehRad51 is weak, it is strongly enhanced by the presence of two HR accessory cofactors, calcium and Hop2-Mnd1. The biochemical system described herein was used to demonstrate the potential for targeting ehRad51 with two small molecule inhibitors of human RAD51. We show that 4,4′-diisothiocyanostilbene-2,2′-disulfonic acid (DIDS) inhibited ehRad51 by interfering with DNA binding and attenuated encystation in Entamoeba invadens, while B02 had no effect on ehRad51 strand exchange activity. These results provide insight into the underlying mechanism of homology-directed DNA repair in E. histolytica