Elective oocyte cryopreservation for age-related fertility decline.

PURPOSE: Women who pursue fertility at an advanced age are increasingly common. Family planning and sexual education have traditionally focused on contraception and prevention of sexually transmitted diseases. A focus should now also be placed on fertility awareness and fertility preservation. This manuscript aims to give an update on the existing evidence around elective oocyte cryopreservation, also highlighting the need for fertility education and evidence-based, individualized counselling. METHODS: A thorough electronic search was performed from the start of databases to March 2020 aiming to summarize the existing evidence around elective egg freezing, the logic behind its use, patient counselling and education, success rates and risks involved, regulation, cost-effectiveness, current status and future perspectives. RESULTS: Clinician-led counselling regarding reproductive aging and fertility preservation is often overlooked. Elective oocyte cryopreservation is not a guarantee of live birth, and the answer regarding cost-effectiveness needs to be individualized. The existing studies on obstetric and perinatal outcomes following the use of egg freezing are, until now, reassuring. Constant monitoring of short-term and long-term outcomes, uniform regulation and evidence-based, individualized counselling is of paramount importance. CONCLUSIONS: Elective oocyte cryopreservation is one of the most controversial aspects of the world of assisted reproduction, and a lot of questions remain unanswered. However, women today do have this option which was not available in the past. Elective oocyte cryopreservation for age-related fertility decline should be incorporated in women's reproductive options to ensure informed decisions and reproductive autonomy.

Detection of mutations in the rpoB gene of rifampicin-resistant Mycobacterium tuberculosis strains inhibiting wild type probe hybridization in the MTBDR plus assay by DNA sequencing directly from clinical specimens.

BACKGROUND: The potential of genetic testing for rapid and accurate diagnosis of drug-resistant Mycobacterium tuberculosis strains is vital for efficient treatment and reduction in dissemination. MTBDR plus assays rapidly detect mutations related to drug resistance and wild type sequences allied with susceptibility. Although these methods are promising, the examination of molecular level performance is essential for improved assay result interpretation and continued diagnostic development. Therefore this study aimed to determine novel mutations that were inhibiting wild type probe hybridization in the Line probe assay by DNA sequencing. Using data collected from Line Probe assay (GenoType MTBDRplus assay) the contribution of absent wild type probe hybridization to the detection of rifampicin resistance was assessed via comparison to a reference standard method i.e. DNA sequencing. RESULTS: Sequence analysis of the rpoB gene of 47 MTB resistant strains from clinical specimens showed that 37 had a single mutation, 9 had double mutations and one had triple mutations in the ropB gene. CONCLUSIONS: The absence of wild type probe hybridization without mutation probe hybridization was mainly the result of the failure of mutation probe hybridization and the result of the novel or rare mutations. Additional probes are necessary to be included in the Line probe assay to improve the detection of rifampicin-resistant Mycobacterium tuberculosis strains.

Rapid detection of drug-resistant Mycobacterium tuberculosis directly from clinical specimens using allele-specific polymerase chain reaction assay.

Background & objectives: Rapid detection of drug resistance in Mycobacterium tuberculosis (MTB) is essential for the efficient control of tuberculosis. Hence, in this study a nested-allele-specific (NAS) PCR, nested multiple allele-specific PCR (NMAS-PCR) and multiple allele-specific (MAS) PCR assays were evaluated that enabled detection of the most common mutations responsible for isoniazid (INH) and rifampicin (RIF) resistance in MTB isolates directly from clinical specimens. Methods: Six pairs of primers, mutated and wild type, were used for the six targets such as codon 516, 526 and 531 of rpoB, codon 315 of katG and C15-T substitution in the promoter region of mabA-inhA using allele-specific (AS) PCR assays (NAS-PCR, NMAS-PCR and MAS-PCR). The performance of AS PCR method was compared with phenotypic drug susceptibility testing (DST). Results: The usefulness of AS PCR assays was evaluated with 391 clinical specimens (251 Acid fast bacilli smear positive and MTB culture positive; 93 smear negative and MTB culture positive; 47 smear positive and MTB culture negative) and 344 MTB culture positive isolates. With culture-based phenotypic DST as a reference standard, the sensitivity and specificity of the NAS-PCR, NMAS-PCR and MAS-PCR assay for drug resistance-related genetic mutation detection were 98.6 and 97.8 per cent for INH, 97.5 and 97.9 per cent for RIF and 98.9 and 100 per cent for multidrug resistance (MDR). Interpretation & conclusions: The performance of AS PCR assays showed that those could be less expensive and technically executable methods for rapid detection of MDR-TB directly from clinical specimens.

Evaluation of the nutrient profile of Trachyspermum ammi L. seed under the influence of nanoparticles during germination.

Trachyspermum ammi L. commonly known as Ajwain is an annual herb belonging to the family Apiaceae. It is enormously grown in Egypt, Iran, Pakistan, Afghanistan, and India as well as European region. Seeds of Ajwain were highly administered by traditional healers and usually employed for different ailments. Nanomaterials are known to have plant growth promoting effects, which could find applications in agriculture. In this study, the nanoparticles (NPs) showed the potential to enhance the primary metabolites when administered during germination. Therefore, nanoparticles elicitation can be used to increase the productivity, nutritional values and metabolite contents in Trachyspermum ammi L. This study aimed to provide new insight of the potential growth promoting effects of the nanoparticles () on plant system. Different concentrations of two nanoparticles, that is, iron pyrite (FeS2) and molybdenum disulphide (MoS2) at three different concentrations of 25ug/ml, 50ug/ml and 75ug/ml were tested on the seeds of Trachyspermum ammi L. The data indicated that nanoparticles enhanced the seedling growth as greener leafs and increased lengths of epicotyl and hypocotyls were seen. These nanoparticles also showed the potential to increase the contents of primary metabolites during germination and the total soluble protein content in seed was increased in nanoparticles-treated seeds as compared to control. The total protein profiling by SDS-PAGE indicated significant differences in number and molecular weights of protein bands upon exposure to nanoparticles.

Differentiation of Mycobacterium tuberculosis complex from non-tubercular mycobacteria by nested multiplex PCR targeting IS6110, MTP40 and 32kD alpha antigen encoding gene fragments.

BACKGROUND: Control of the global burden of tuberculosis is obstructed due to lack of simple, rapid and cost effective diagnostic techniques that can be used in resource poor-settings. To facilitate the early diagnosis of TB directly from clinical specimens, we have standardized and validated the use of nested multiplex PCR, targeting gene fragments IS6110, MTP40 and 32kD α-antigen encoding genes specific for Mycobacterium tuberculosis complex and non-tubercular mycobacteria (NTM), in comparison to smear microscopy, solid culture and single step multiplex PCR. The results were evaluated in comparison to a composite reference standard (CRS) comprising of microbiological results (smear and culture), clinical, radiological and cytopathological findings, clinical treatment and response to anti-tubercular therapy. METHODS: The nested multiplex PCR (nMPCR) assay was evaluated to test its utility in 600 (535 pulmonary and 65 extra-pulmonary specimens) clinically suspected TB cases. All specimens were processed for smear, culture, single step multiplex PCR and nested multiplex PCR testing. RESULTS: Out of 535 screened pulmonary and 65 extra-pulmonary specimens, 329 (61.5%) and 19 (29.2%) cases were culture positive for M. tuberculosis. Based on CRS, 450 patients had "clinical TB" (definitive-TB, probable-TB and possible-TB). Remaining 150 were confirmed "non-TB" cases. For culture, the sensitivity was low, 79.3% for pulmonary and 54.3% for extra-pulmonary cases. The sensitivity and specificity results for nMPCR test were evaluated taken composite reference standard as a gold standard. The sensitivity of the nMPCR assay was 97.1% for pulmonary and 91.4% for extra-pulmonary TB cases with specificity of 100% and 93.3% respectively. CONCLUSION: Nested multiplex PCR using three gene primers is a rapid, reliable and highly sensitive and specific diagnostic technique for the detection and differentiation of M. tuberculosis complex from NTM genome and will be useful in diagnosing paucibacillary samples. Nested multiplex PCR assay was found to be better than single step multiplex PCR for assessing the diagnosis of TB.

Tuning phonon properties in thermoelectric materials.

This review article presents a discussion of theoretical progress made over the past several decades towards our understanding of thermoelectric properties of materials. Particular emphasis is placed upon describing recent progress in 'tuning' phonon properties of nanocomposite materials for gaining enhancement of the thermoelectric figure of merit.

Identification of pathologically insignificant prostate cancer is not accurate in unscreened men.

BACKGROUND: Identification of men harbouring insignificant prostate cancer (PC) is important in selecting patients for active surveillance. Tools have been developed in PSA-screened populations to identify such men based on clinical and biopsy parameters. METHODS: Prospectively collected case series of 848 patients was treated with radical prostatectomy between July 2007 and October 2011 at an English tertiary care centre. Tumour volume was assessed by pathological examination. For each tool, receiver operator characteristics were calculated for predicting insignificant disease by three different criteria and the area under each curve compared. Comparison of accuracy in screened and unscreened populations was performed. RESULTS: Of 848 patients, 415 had Gleason 3+3 disease on biopsy. Of these, 32.0% had extra-prostatic extension and 50.2% were upgraded. One had positive lymph nodes. Two hundred and six (24% of cohort) were D'Amico low risk. Of these, 143 had more than two biopsy cores involved. None of the tools evaluated has adequate discriminative power in predicting insignificant tumour burden. Accuracy is low in PSA-screened and -unscreened populations. CONCLUSIONS: In our unscreened population, tools designed to identify insignificant PC are inaccurate. Detection of a wider size range of prostate tumours in the unscreened may contribute to relative inaccuracy.

Adalimumab (tumor necrosis factor-blocker) reduces the expression of glial fibrillary acidic protein immunoreactivity increased by exogenous tumor necrosis factor alpha in an organotypic culture of porcine neuroretina.

PURPOSE: To determine if exogenous addition of tumor necrosis factor alpha (TNFα) exacerbates retinal reactive gliosis in an organotypic culture of porcine neuroretina and to evaluate if concomitant adalimumab, a TNF-blocker, diminishes it. METHODS: Porcine retinal explants from 20 eyeballs were cultured. Cultures with 100 pg/ml TNFα, 10 µg/ml adalimumab, 100 pg/ml TNFα plus 10 µg/ml adalimumab, or controls without additives were maintained for 9 days. Freshly detached retinas were processed in parallel. TNFα levels in control culture supernatants were quantified with enzyme-linked immunosorbent assay. Cryostat sections were doubly immunostained for glial fibrillary acidic protein (GFAP), a marker for reactive gliosis, and cellular retinaldehyde-binding protein (CRALBP), a marker for Müller cells. Sections were also labeled with the isolectin IB4, a label for microglia/macrophages. RESULTS: TNFα in control culture supernatants was detected only at day 1. Compared to the fresh neuroretinal samples, upregulation of GFAP and downregulation of CRALBP occurred during the 9 days of culture. Exogenous TNFα stimulated glial cells to upregulate GFAP and downregulate CRALBP immunoreactivity. TNFα-treated cultures also initiated the growth of gliotic membranes and underwent retinal disorganization. Adalimumab inhibited the spontaneous increases in GFAP and maintained CRALBP. In combination with TNFα, adalimumab reduced GFAP expression and conserved CRALBP, with only slight retinal disorganization. No appreciable changes in IB4 labeling were observed under the different culture conditions. CONCLUSIONS: In cultured porcine neuroretina, spontaneous reactive gliosis and retinal disorganization were exacerbated by exogenous TNFα. Adalimumab reduced spontaneous changes and those induced by TNFα. Therefore, inhibiting TNFα may represent a novel approach to controlling retinal fibrosis observed in some human diseases.

Identification of novel inhibitors targeting TIRAP interactions with BTK and PKCδ in inflammation through an in silico approach.

Advanced computational tools focusing on protein-protein interaction (PPI) based drug development is a powerful platform to accelerate the therapeutic development of small lead molecules and repurposed drugs. Toll/interleukin-1 receptor (TIR) domain-containing adapter protein (TIRAP) and its interactions with other proteins in macrophages signalling are crucial components of severe or persistent inflammation. TIRAP activation through Bruton's tyrosine kinase (BTK) and Protein Kinase C delta (PKCδ) is essential for downstream inflammatory signalling. We created homology-based structural models of BTK and PKCδ in MODELLER 9.24. TIRAP interactions with BTK and PKCδ in its non-phosphorylated and phosphorylated states were determined by multiple docking tools including HADDOCK 2.4, pyDockWEB and ClusPro 2.0. Food and Drug Administration (FDA)-approved drugs were virtually screened through Discovery Studio LibDock and Autodock Vina tools to target the common TIR domain residues of TIRAP, which interact with both BTK and PKC at the identified interfacial sites of the complexes. Four FDA-approved drugs were identified and found to have stable interactions over a range of 100 ns MD simulation timescales. These drugs block the interactions of both kinases with TIRAP in silico. Hence, these drugs have the potential to dampen downstream inflammatory signalling and inflammation-mediated disease.

Ripening of tomato (Solanum lycopersicum L.). Part II: Regulation by its stem scar region.

The relationship between ripening behaviour and stem scar region of fruit in different tomato varieties (stored at 30.5 ± 1°C or 25 ± 1°C) was studied by blocking the stem scar region either completely or to different extent with high vacuum silicone grease. In 'Pusa Ruby' (a fast ripening variety) and 'Pusa Gaurav' (slow ripening variety), complete blockage of stem scar region of fruits at green mature stage showed 3-fold reduction in ripening index at 14 days after treatment (DAT) but with increased rottage. It was demonstrated in 'Pusa Ruby' and 'Pusa 120' that the extent of blockage of stem scar region has dependence on the rate of respiration and ripening suppression. The extent of climacteric rise was reduced significantly in the treated fruits. These suppressive effects of treatment were found to diminish with the advancement in the ripening stage of tomato fruit.

Anharmonic, dimensionality and size effects in phonon transport (2017J. Phys.: Condens. Matter29 505703).

Corrigendum to 2017J. Phys.: Condens. Matter29505703.

Tunable Thermal Transport Characteristics of Nanocomposites.

We present a study of tunable thermal transport characteristics of nanocomposites by employing a combination of a full-scale semi-ab inito approach and a generalised and extended modification of the effective medium theory. Investigations are made for planar superlattices (PSLs) and nanodot superlattices (NDSLs) constructed from isotropic conductivity covalent materials Si and Ge, and NDSLs constructed from anisotropic conductivity covalent-van der Waals materials MoS 2 and WS 2 . It is found that difference in the conductivities of individual materials, period size, volume fraction of insertion, and atomic-level interface quality are the four main parameters to control phonon transport in nanocomposite structures. It is argued that the relative importance of these parameters is system dependent. The equal-layer thickness Si/Ge PSL shows a minimum in the room temperature conductivity for the period size of around 4 nm, and with a moderate amount of interface mass smudging this value lies below the conductivity of SiGe alloy.

Frequency and degree of inter-trait association of maxillary Non-Metric Dental Crown Traits in the permanent dentitions of two states of India.

Non-metric Dental Crown Traits are a principal source of information in forensic dentistry. However, inadequate data on the prevalence of these traits prompted this study to determine the frequency, sexual dimorphism and degree of inter-trait association in two different populations of India. Dichotomized data on the existence of non-metric features were recorded among individuals from Odisha (n=506) and Kerala (n=536) between 15 to 30 years of age. Cusp of Carabelli is the most common trait to occur (48 %) followed by shovelling of incisors (15%) and Bushman canine (14%). Bushman canine (p=0.045) and Cusp of Carabelli (p = 0.041) were found to be significantly expressed in Odisha and Kerala populations respectively. A strong association between shovelling of central incisor and Bushman canine with a likelihood ratio of 14.041 (p=0.001) was observed. This study will help in characterizing the Indian dentition and post-mortem dental profiling.

The anharmonic phonon decay rate in group-III nitrides.

Measured lifetimes of hot phonons in group-III nitrides have been explained theoretically by considering three-phonon anharmonic interaction processes. The basic ingredients of the theory include full phonon dispersion relations obtained from the application of an adiabatic bond charge model and crystal anharmonic potential within the isotropic elastic continuum model. The role of various decay routes, such as Klemens, Ridley, Vallée-Bogani and Barman-Srivastava channels, in determining the lifetimes of the Raman active zone-centre longitudinal optical (LO) modes in BN (zincblende structure) and A(1)(LO) modes in AlN, GaN and InN (wurtzite structure) has been quantified.

Quantitative study of the enhancement of the thermal conductivity of AlN ceramics by nanoscale processing.

We have theoretically studied and quantitatively analysed the enhancement of the thermal conductivity of AlN microceramics (i.e. commercially available powder) by the insertion of AlN nanosized particles of high-specific surface area, and of Y(2)O(3) and CaO as sintering additives. We have also studied the enhancement of the thermal conductivity of AlN nanoceramics by using Y(2)O(3) as the sintering additive. Thermal conductivity calculations have been carried out by applying the Callaway theory in its full form and by incorporating a detailed and accurate account of three-phonon processes. The role of densification of the AlN ceramic samples in enhancing the thermal conductivity has been quantified at low, intermediate and high temperatures. In addition to explaining the experimentally observed room-temperature conductivity results, the conductivity variation has been predicted over a large temperature range.

Mode confinement, interface mass-smudging, and sample length effects on phonon transport in thin nanocomposite superlattices.

We employ a semi-ab initio theoretical method to investigate mode confinement, interface mass-smudging, and sample length effects on phonon transport in thin nanocomposite superlattices. We present a detailed comparative study of numerical results showing the reduction in thermal conductivity due to each of these three effects for Si/Ge nanocomposite structures with planar superlattice (SL), embedded nanowire superlattice (NWSL), and embedded nanodot superlattice (NDSL) geometries. Importantly, it is found that any of these three types of thin period systems, with small amounts of interface mass smudging, can exhibit a room-temperature conductivity significantly lower than the SiGe alloy conductivity, providing strong evidence that they could be used as efficient thermoelectric materials. It is also found that the room-temperature conductivity of each of the nanocomposite superlattices shows a weaker sample size dependence than do the component bulk conductivities.

Should vitamin D be routinely checked for all chronic obstructive pulmonary disease patients?

AIMS AND OBJECTIVES: This study aimed to compare the vitamin D levels between chronic obstructive pulmonary disease (COPD) patients and healthy controls and to describe the correlation between vitamin D levels and lung functions. METHODS: Fifty COPD patients (cases) and 30 healthy volunteers (controls) were recruited and their serum vitamin D level was measured together with lung function (forced vital capacity and forced expiratory volume in 1 s [FEV1]) by spirometry. vitamin D was categorized as ≤20 nmol/l: deficient, 21-50 nmol/l: inadequate, and ≥51 nmol/l as sufficient. RESULTS: In this case-control cross-sectional study, lower vitamin D levels were associated with lower lung function in both cases as well as controls, the effect being more pronounced in cases. Mean FEV1 at vitamin D ≤20 nmol/l (0.98 ± 0.40 vs. controls 1.93 ± 0.24 with P = 0.006), mean FEV1 at vitamin D 21-50 nmol/l (1.55 ± 0.54 vs. 2.20 ± 0.31 with P = 0.000), and mean FEV1 at vitamin D ≥51 nmol/l (2.06 ± 0.54 vs. 2.20 ± 0.31 with P = 0.002). Moreover, the severity of predicted postbronchodilator FEV1% was also much lower among COPD cohort versus healthy volunteers (mean FEV1%: cases 47.88 ± 14.22 vs. controls 58.76 ± 15.05 with P = 0.002). CONCLUSIONS: Importantly, lung function in both the groups was affected by decreased vitamin D level; decrease in FEV1 was more pronounced among COPD patients compared to controls showing more expiratory airflow limitation. Vitamin D levels are associated with changes in lung function in cases of COPD as well as healthy controls. Larger studies to confirm the association in Indian context are required and routine assessment of vitamin D may be undertaken to obviate the effects of low vitmain D level on lung function.

No high Tibetan Plateau until the Neogene.

The Late Paleogene surface height and paleoenvironment for the core area of the Qinghai-Tibetan Plateau (QTP) remain critically unresolved. Here, we report the discovery of the youngest well-preserved fossil palm leaves from Tibet. They were recovered from the Late Paleogene (Chattian), ca. 25.5 ± 0.5 million years, paleolake sediments within the Lunpola Basin (32.033°N, 89.767°E), central QTP at a present elevation of 4655 m. The anatomy of palms renders them intrinsically susceptible to freezing, imposing upper bounds on their latitudinal and altitudinal distribution. Combined with model-determined paleoterrestrial lapse rates, this shows that a high plateau cannot have existed in the core of Tibet in the Paleogene. Instead, a deep paleovalley, whose floor was <2.3 km above mean sea level bounded by (>4 km) high mountain systems, formed a topographically highly varied landscape. This finding challenges prevailing views on tectonic processes, monsoon dynamics, and the evolution of Asian biodiversity.

The major 8p22 tumor suppressor DLC1 is frequently silenced by methylation in both endemic and sporadic nasopharyngeal, esophageal, and cervical carcinomas, and inhibits tumor cell colony formation.

Identification of tumor suppressor genes (TSG) silenced by methylation uncovers mechanisms of tumorigenesis and identifies new epigenetic tumor markers for early cancer detection. Both nasopharyngeal carcinoma (NPC) and esophageal carcinoma are major tumors in Southern China and Southeast Asia. Through expression subtraction of NPC, we identified Deleted in Liver Cancer 1 (DLC1)/ARHGAP7 (NM_006094)--an 8p22 TSG as a major downregulated gene. Although expressed in all normal tissues, DLC1 was silenced or downregulated in 11/12 (91%) NPC, 6/15 (40%) esophageal, 5/8 (63%) cervical and 3/9 (33%) breast carcinoma cell lines. No genetic deletion of DLC1 was detected in NPC although a hemizygous deletion at 8p22-11 was found by 1-Mb array-CGH in some cell lines. We then located the functional DLC1 promoter by 5'-RACE and promoter activity assays. This promoter was frequently methylated in all downregulated cell lines and in a large collection of primary tumors including 89% (64/72) NPC (endemic and sporadic types), 51% (48/94) esophageal, 87% (7/8) cervical and 36% (5/14) breast carcinomas, but seldom in paired surgical marginal tissues and not in any normal epithelial tissue. The transcriptional silencing of DLC1 could be reversed by 5-aza-2'-deoxycytidine or genetic double knock-out of DNMT1 and DNMT3B. Furthermore, ectopic expression of DLC1 in NPC and esophageal carcinoma cells strongly inhibited their colony formation. We thus found frequent epigenetic silencing of DLC1 in NPC, esophageal and cervical carcinomas, and a high correlation of methylation with its downregulation, suggesting a predominant role of epigenetic inactivation. DLC1 appears to be a major TSG implicated in the pathogenesis of these tumors, and should be further tested as a molecular biomarker in patients with these cancers.

Identification of a tumor suppressive critical region mapping to 3p14.2 in esophageal squamous cell carcinoma and studies of a candidate tumor suppressor gene, ADAMTS9.

A gene critical to esophageal cancer has been identified. Functional studies using microcell-mediated chromosome transfer of intact and truncated donor chromosomes 3 into an esophageal cancer cell line and nude mouse tumorigenicity assays were used to identify a 1.61 Mb tumor suppressive critical region (CR) mapping to chromosome 3p14.2. This CR is bounded by D3S1600 and D3S1285 microsatellite markers. One candidate tumor suppressor gene, ADAMTS9, maps to this CR. Further studies showed normal expression levels of this gene in tumor-suppressed microcell hybrids, levels that were much higher than observed in the recipient cells. Complete loss or downregulation of ADAMTS9 gene expression was found in 15 out of 16 esophageal carcinoma cell lines. Promoter hypermethylation was detected in the cell lines that do not express this gene. Re-expression of ADAMTS9 was observed after demethylation drug treatment, confirming that hypermethylation is involved in gene downregulation. Downregulation of ADAMTS9 was also found in 43.5 and 47.6% of primary esophageal tumor tissues from Hong Kong and from the high-risk region of Henan, respectively. Thus, this study identifies and provides functional evidence for a CR associated with tumor suppression on 3p14.2 and provides the first evidence that ADAMTS9, mapping to this region, may contribute to esophageal cancer development.