Drosophila and C. elegans.Sexual differentiation often has significant effects on life span and aging phenotypes. For example, males and females of several species have different life spans, and genetic and environmental manipulations that affect life span often have different magnitude of effect in males versus females. Moreover, the presence of a differentiated germ-line has been shown to affect life span in several species, including Drosophila melanogaster. Drosophila females heterozygous for the tudor[1] mutation produce normal offspring, while their homozygous sisters produce offspring that lack a germ line. To identify additional sexual differentiation genes that might affect life span, the conditional transgenic system Geneswitch was employed, whereby feeding adult flies or developing larvae the drug RU486 causes the over-expression of selected UAS-transgenes.Experiments were conducted to determine how alterations in sexual differentiation gene activity might affect the life span of tudor[1] mutation was examined in a long-lived genetic background, and was found to increase life span in males but not in females, consistent with previous reports. Fitting the data to a Gompertz-Makeham model indicated that the maternal tudor[1] mutation increases the life span of male progeny by decreasing age-independent mortality. The Geneswitch system was used to screen through several UAS-type and EP-type P element mutations in genes that regulate sexual differentiation, to determine if additional sex-specific effects on life span would be obtained. Conditional over-expression of transformer female isoform (traF) during development produced male adults with inhibited sexual differentiation, however this caused no significant change in life span. Over-expression of doublesex female isoform (dsxF) during development was lethal to males, and produced a limited number of female escapers, whereas over-expression of dsxF specifically in adults greatly reduced both male and female life span. Similarly, over-expression of fruitless male isoform A (fru-MA) during development was lethal to both males and females, whereas over-expression of fru-MA in adults greatly reduced both male and female life span.In this study germ-line ablation caused by the maternal Manipulation of sexual differentiation gene expression specifically in the adult, after morphological sexual differentiation is complete, was still able to affect life span. In addition, by manipulating gene expression during development, it was possible to significantly alter morphological sexual differentiation without a significant effect on adult life span. The data demonstrate that manipulation of sexual differentiation pathway genes either during development or in adults can affect adult life span. C. elegans, elimination of reproduction by ablation of the germ line extended life span by up to +60%  mutation was tested in a particularly long-lived genetic background, and was found to increase the life span of male offspring, but to have neutral or negative effects on female life span.In  females . The eliffspring . Interesife span , and theDrosophila soma, sexual differentiation is determined by the X chromosome to autosome (X:A) ratio and the on/off status of the master regulatory gene Sex-lethal (Sxl)  Figure , howeverthe soma -22. GeneDrosophila larval development, or specifically in the adult stages. The Drosophila sex determination hierarchy consists of pre-mRNA splicing factors encoded by the genes Sex-lethal (Sxl), transformer (tra), and transformer-2 (tra-2)  Figure . The tiscterized ,31. Act-Drosophila culture and life span assays were performed essentially as previously described  males were also crossed to Act-GS-255B virgins, and the progeny were cultured on food supplemented with drug (+RU486) or with ethanol only, as the controls.UAS-p value were calculated using R 2.6.2. Analysis of mortality rate was performed using the WinModest statistical package  mutation were crossed to Oregon-R wild-type male flies to produce control offspring and offspring lacking the germ line, respectively , while the initial mortality rate (rate constant a) and mortality rate increase with time (rate constant b) were not significantly affected  during development was lethal, and over-expression of fru MA in adults greatly reduced both male and female mean life span, by -61.97% and -64.5%, respectively , when expressed during development or in adults , whereas female life span was not affected. In a previous report, germ line ablation using the tudor[1] mutation increased the median life span of male flies by +8.6% , and the control males and control females had median life spans of 82 days and 80 days, respectively  gene in adult flies, which caused full sterility by day 7 post-eclosion in females, and GC depopulation in the 3rd instar larval (L3) stage or later in males. The presence of a complete germ line in the earlier stages of development followed by GC loss at a later stage could affect life span in a different way, compared with the lack of a germ line from the beginning of embryogenesis, such as is produced by the tudor mutation. For example, the tudor mutation also partially inhibits formation of the somatic gonad, while ablation of GCs using bam mis-expression at later developmental stages allows for complete differentiation of the somatic gonad. Consistent with this idea, in C. elegans, extension of life span by ablation of the germ line requires the presence of the somatic gonad  mutant data to a Gompertz-Makeham model, which allowed us to separate early mortality from the age-dependent and age-independent mortality. The analysis indicated that germ line ablation in male Drosophila extended life span by decreasing the age-independent mortality. That implies that germ line ablation provides a benefit for survival in male flies, and this beneficial effect is constant over the adult life span. This might occur through altered IIS as reported for C. elegans hermaphrodites [The hrodites .tra during development significantly inhibited sexual differentiation in males, yet produced no significant change in adult life span. Expression of tra in males has previously been found to feminize locomotor activity patterns [The other manipulations of the sexual differentiation pathway tested produced either neutral or negative effects on adult fly life span. For example, conditional over-expression of patterns . These ddsxF during development was lethal to males and produced a limited number of females, while over-expression of dsxF in adults greatly reduced both male and female life span. Over-expression of dsxM during development was toxic to males and females, whereas over-expression of dsxM in adults produced only a small, if any, reduction in male and female life span. This indicates that in adults, where sexual differentiation is already complete, changes in expression of specific sex-determination pathway genes are still able to have significant effects on life span.Over-expression of fru male isoform A (fruMA) during development was lethal, and over-expression of fruMA in adults greatly reduced both male and female mean life span. However, expression of an RNAi construct specific for fruMA during development or in adults significantly decreased female life span, but did not give a consistent change in male life span. The reason for this effect of fruMA-RNAi in females is not yet clear. In females, fru P1 transcripts are produced, but not translated [fruMA-RNAi could still function through fru P1 transcripts to affect female life span, or could function through targets other than fru.Interestingly, over-expression of anslated . PossiblfruMA and dsxF shortened life span to a similar extent in males and females. This may be because the sex-specificity of these factors is created during development by means of sex-specific expression, and in our experiments we are forcing expression in each sex, thereby eliminating sex-specificity of action. One possible exception is the developmental lethality caused by over-expression of dsxF during larval development, which was complete in males, where it is not normally expressed, and only partial in females, where it is normally present.Taken together with life span results of other assayed genes, our data show little correspondence between the sex-specific mode of action of sexual differentiation genes in development and sex-specific effects of their over-expression or knockdown on adult lifespan. For example, adult-specific over-expression of fru is essential to generate several male-specific behaviors, including male courtship behavior [Sexual differentiation genes play important roles in germ cell development -22. Therbehavior . Althougbehavior ,10,11.The data demonstrate that manipulation of sexual differentiation gene expression specifically in the adult, after morphological sexual differentiation is complete, is able to affect life span. In addition, by manipulating gene expression during development, it was possible to significantly alter morphological sexual differentiation without a significant effect on adult life span. The data demonstrate that manipulation of sexual differentiation pathway genes either during development or in adults can affect adult life span, sometimes with sex-specific effects, and suggest it should be possible in the future to investigate how the sexual differentiation pathway interacts with specific life span regulatory genes to produce sex-specific differences in longevity.The authors declare that they have no competing interests.tudor mutant offspring. JS and GL conducted experiments with gene over-expression and RNAi. JS performed statistical analyses, with help from DF and GL. JT conceived of and supervised the project, and JT and JS wrote the paper. All authors have read and approved the final manuscript.DF conducted life span assay of The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2318/9/56/prepub


Cancer of the esophagus is a deadly malignancy, and development of biomarkers that predict survival is an urgent need. The apoptotic pathways have been hypothesized as important in progression of esophageal squamous cell carcinoma (ESCC). We investigated a panel of proteins that regulate apoptosis as candidate of biomarkers of prognosis in ESCC.Tissue microarray (TMA) including 313 surgically-resected cases of ESCC specimens was built for immunohistochemical interrogation. We evaluated seven genes in the FasL-Fas apoptotic pathway - FasL, Fas, FAS-associated death domain protein (FADD), phosphorylated-FADD, and caspase 8 and 10, and the antiapoptotic protein bcl-2. We studied pathway integrity and relations to risk and clinical factors, and determined the prognostic significance of each marker.Five markers showed strong inter-marker correlations , including FasL, Fas, FADD, and caspases 8 and 10. FasL and FADD also showed modest correlations with one or more cancer risk factors, but none of the markers was significantly associated with either tumor stage or lymph node metastasis, the only two clinical factors that predicted survival in these ESCC cases. Multivariate-adjusted proportional hazard regression models showed no association between protein expression and risk of death for any of the seven markers examined.Individual biomarkers in the apoptosis pathway do not appear to predict survival of patients with ESCC. FADD) and under-expression of Fas and caspase 8  on cases using a structured interview with a questionnaire administered by a nurse in the hospital prior to surgery. Clinical data was abstracted from hospital records after surgery. Demographic, lifestyle and clinical data for cases included in this study are shown in Table Details of the TMA construction were previously described . In brieThe TMA sections were stained with antibodies to FasL , Fas , FADD , p-FADD , caspase 8 , caspase 10 , and bcl-2 . Slides were stained according to manufacturer's protocols. The immunohistochemical staining patterns were validated against previously described patterns of staining for each marker on a separate TMA, including appropriate positive and negative controls. Internal positive and negative controls, including normal squamous epithelium of the esophagus from non-cancer patients were utilized as available to further support the staining patterns.Fas and FasL was expressed exclusively in cell membranes, whereas immunoreactivity of FADD and bcl-2 was cytoplasmic. Caspase 8 and 10 expression was detected in both cytoplasm and nucleus. p-FADD was primarily expressed in the nucleus of cells. Staining results were scored based on: (i) percent of positive tumor cells in tumor tissue: zero (0%), 1 (1-25%), 2 (26-50%), 3 (51-75%) and 4 (76-100%); and (ii) signal intensity: zero , 1 (weak), 2 (moderate) and 3 (marked). An overall score was calculated by multiplying the percent cells positive score by the intensity score (range zero to 12).P-values were two-sided and considered statistically significant if P < 0.05.All statistical analyses were performed using Statistical Analysis Systems (SAS) . Spearman correlation coefficients were used to assess associations between the seven different apoptosis biomarkers, and between the seven markers and risk factors/clinico-pathologic features. Overall survival time was calculated as the date of surgery to the date of death or the date last known alive. Survival was examined graphically with Kaplan-Meier curves and analyzed statistically with log-rank tests and proportional hazards regression models (SAS PHREG procedure) adjusted for lifestyle and tumor characteristics as covariates as previously described . All P-vOverall protein expression for the seven markers evaluated here showed low levels of positivity, ranging from zero to 15% positive Table . The SpeWe successfully contacted 261 of 265 ESCC cases or their families during follow-up for vital status; 181 cases died and 80 were still alive at the end of follow-up. Median overall survival was 677 days  and 15 cases were still alive five or more years post-surgery; the longest survivor was still alive six years and two months after surgery. Kaplan-Meier graphs and log-rank analyses of individual markers did not reveal differences in overall survival by protein expression positivity (data not shown). Analyses adjusted for demographic attributes and potential confounding factors in Cox proportional hazard regression models also failed to identify significant associations between markers and survival time /BAK (Bcl-2-antagonist/killer1) proteins, which induce the permeabilization of the outer mitochondrial membrane, a crucial step for apoptotic cell death . Caspaseet al. also previously reported that Fas and FasL were not related to disease-free survival in ESCC [et al. observed significantly longer ESCC-free survival in patients with Fas-positive (versus Fas-negative) tumors, and in FasL-negative (versus FasL-positive) tumors [et al. reported that Fas expression was an independent prognosticator for recurrence-free survival, but that FasL expression did not influence ESCC survival.[In accord with our findings, Xue  in ESCC . In cont) tumors . Shibakisurvival. Studies survival.. There aet al [et al. reported that FADD expression correlated with decreased survival in ESCC [et al. found that bcl-2 expression correlated with better survival, and was an independent prognostic factor after multivariate analysis[et al. and Ohbu et al., however, in both studies bcl-2 was not an independent prognostic value after adjustment for other variables in multivariate analysis [FADD did not predict survival in ESCC in our study, a finding similar to that of Chang et al . In cont in ESCC . Inducti in ESCC . Consist in ESCC ,14. Chan analysis. Similaranalysis ,16.FasL expression showed significant correlation with three risk factors including male sex, tobacco use, and family history of UGI cancer Table . Fas hadWhile the current study evaluated only protein expression in relation to survival and not the potential use of these biomarkers in the early detection of ESCC, the generally low prevalence of expression positivity indicates that they would not be suitable candidates for early detection markers. We were unable to identify a role for biomarkers in the FasL-Fas apoptotic pathway and prognosis in ESCC.The authors declare that they have no competing interests.NH, PRT, and SMH designed the study. NH, JZS, and QHW were responsible for management of patients' data and tissues. MT and SMH performed IHC and provided the scoring data. CG, NH, and PRT performed statistical analysis. MT and SMH wrote the manuscript, and NH and PRT helped editing the manuscript. All authors read and approved the final version of the manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2407/9/310/prepub


Hazardous health effects stemming from exposure to radiofrequency electromagnetic waves (RF-EMW) emitted from cell phones have been reported in the literature. However, the cellular target of RF-EMW is still controversial. This review identifies the plasma membrane as a target of RF-EMW. In addition, the effects of RF-EMW on plasma membrane structures  and voltage-gated calcium channels are discussed. We explore the disturbance in reactive oxygen species (ROS) metabolism caused by RF-EMW and delineate NADH oxidase mediated ROS formation as playing a central role in oxidative stress (OS) due to cell phone radiation . This review also addresses: 1) the controversial effects of RF-EMW on mammalian cells and sperm DNA as well as its effect on apoptosis, 2) epidemiological, in vivo animal and in vitro studies on the effect of RF-EMW on male reproductive system, and 3) finally, exposure assessment and dosimetry by computational biomodeling. The health effects of radiofrequency electromagnetic waves (RF-EMW) emitted from cell phones have been debated greatly ,2. ReseaRecent studies demonstrated microwaves emitted from the cell phone, i.e., RF-EMW do not produce thermal effect at specific absorption rate (SAR) of 1.6 Watts/Kg -7. HowevAmong the effect of RF-EMW on various body organs, effect of RF-EMW on brain is the most researched area -24. AddiIn this article, we review the effects of RF-EMW on free radical metabolism and carcinogenesis as well as the epidemiological, in vitro animal and in vitro human studies that have assessed the effect of RF-EMW on male fertility. We also briefly discuss the novel computational biomodeling for in vitro study on human semen currently being performed at our center.et al suggested that RF-EMW may affect the plasma membrane signal transduction, independent of thermal effect [et al later showed that exposure to 900 MHz RF-EMW was associated with a significant increase in annexin V-positive human lymphocytes (without any changes in apoptotic cells) and in the mitochondrial membrane potential [The literature contains controversial reports on the effects of RF-EMW on various cellular organelles. Of particular note, researchers have consistently demonstrated that RF-EMW affects the plasma membrane in cells ,11,12,26otential -34. Thiset al. reported that RF-EMW stimulates plasma membrane NADH oxidase (of HeLa cells) and causes production of extracellular superoxide [Recently, Friedman peroxide . This efRao et al recently provided new evidence supporting the theory that RF-EMW affects the plasma membrane . They stThese studies suggest that the plasma membrane might be the target of RF-EMW and that other observed effects of RF-EMW might be secondary to the effect on the plasma membrane.in vivo animal studies have shown that OS develops in response to cell phone radiation [Free radicals are a group of highly reactive molecules consisting of unpaired electrons in the outer orbit. Free radicals that are derived from oxygen metabolism are known as reactive oxygen species (ROS) . ROS areadiation -42. RF-Eadiation -42. Chroadiation ,41,43-45adiation ,47.in vitro studies. As a consequence of increased levels of free radicals, various cellular and physiological processes can be affected including gene expression, release of calcium from intracellular storage sites, cell growth, and apoptosis.We suggest that RF-EMW induces NADH oxidase enzyme stimulation , which mThe effects of RF-EMW on DNA damage have been reported in various studies in the last decade ,48-51. LLai and Singh demonstrated that treating rats with free radical scavengers blocked the effects of EMW on DNA . This suet al reported that human endothelial cells showed changes in the expression of several genes after exposure to 900 MHZ but not 1800 MHz [et al reported that 1950 MHz RF-EMW exposure for 24 hour at a SAR of 0.05 W/kg induced genotoxic effects in vitro in human fibroblasts but not in lymphocytes [However, the results of more recent studies are controversial regarding the effects of RF-EMW on DNA ,51,58-611800 MHz . Schwarzphocytes . In a stphocytes . On the phocytes . Recent phocytes ,65. Thusphocytes .DNA damage in cells may have an important implication as it is cumulative. Normally, DNA is capable of repairing itself. Through a homeostatic mechanism, cells maintain a delicate balance between DNA damage and repair. DNA damage accumulates if this balance is altered. Most cells can repair single-strand DNA breaks. However, DNA double strand breaks, if not properly repaired, are known to lead to cell death or apoptosis.It has been suggested that RF-EMW might affect the apoptosis pathway. However, the effect of RF-EMW on apoptosis is also controversial ,28,41,67A number of recent reports have suggested a possible link between cell phone use and male infertility ,46,68,69et al. studied 371 men undergoing infertility evaluations and reported that the duration of possession and the daily transmission time of cell phones correlated negatively with the proportion of rapidly progressive motile spermatozoa, suggesting that prolonged use of cell phones might have negative effects on sperm motility [et al. (2002), in a small prospective study involving 13 men with normal semen analysis, also found that using cell phones for 6 h a day for 5 days decreased the rapid progressive motility of spermatozoa [in vivo human exposure study to investigate the effects of cell phone radiation on semen parameters is not feasible due to ethical issues.An initial study from our group involving 361 men who had attended an infertility clinic suggested that the use of cell phones adversely affects semen quality by decreasing sperm count, motility, viability and morphology, which might contribute to male infertility. Similarmotility . Davoudiet al. demonstrated a decrease in seminiferous tubular diameter and epithelium thickness after applying RF-EMW of 869 to 894 MHz [2 at a frequency of 2.45 GHz for 30-40 minutes resulted in significant degeneration of the seminiferous epithelium in mice [In addition to the epidemiological studies, the effects of RF-EMW are well studied in animal studies and "in vitro" studies on human semen. Many studies have indicated that EMW decreases the size of the testicular organs. A decrease in the diameter of the seminiferous tubules ,44 has b 894 MHz . These r in mice . However in mice ,45. Wang in mice . RF-EMW- in mice .Previously, we discussed that RF-EMW can lead to OS. Human spermatozoa are highly susceptible to OS due to high contents of poly unsaturated fatty acids (PUFA) in their cell membrane . RF-EMW 2+ [2+[PKC is a multifunctional protein kinase and is structurally a group of proteins with at least 10 isoenzymes identified thus far . PKC iso2+ ,83. In f2+ [2+PKC is highly concentrated in the equatorial segment, suggesting a specific compartmentalized role for PKC in human sperm physiology. Studies have reported declines in sperm motility with decreases in PKC activity ,84. ReceRF-EMW may also alter intracellular calcium homeostasis by acting on plasma membrane calcium channels . Rao et Previous reports have suggested that electromagnetic fields alter the proliferation rate of cells as well as the rate of DNA, RNA, and protein synthesis ,90. HoweWe suggest the possible pathways of carcinogenesis by RF-EMW.As we have discussed, the plasma membrane may be a target of RF-EMW. RF-EMW might trigger uncontrolled cell proliferation by its action on various plasma membrane enzymes and receptors. Short-term exposure to RF-EMW can lead to increase in the activity of plasma membrane NADH oxidase enzyme, which increases ROS formation . The ROSMoreover, various reports have suggested that ODC may be a target for microwave radiations -95. ODC Although RF-EMW exposure is known to affect PKC isoenzyme expression, the literature contains controversial reports regarding the role of change in PKC activities in carcinogenesis ,97. PhorChanges in intracellular calcium levels and activities of ODC and PKC are interrelated as well as can be secondary to RF-EMW exposure. The literature also contains a report of ROS-mediated changes in the activities of PKC and ODC . Thus, Rin vivo.Carrying a cell phone in a pant pocket (or cell phone clipped to a belt on the waist) exposes the testes to high-power density mobile radiation. Similarly, this occurs when a person is talking on a Bluetooth headset (or any other hands-free accessories) with the cell phone in the pant pocket. Recent use of hands-free accessories might decrease radiation exposure to the human head  but it mAs per WHO research agenda for radiofrequency fields, in vitro studies are important for the health risk assessment due to RF-EMW . We haveWe have reviewed the literature to better understand the effects of cell phone radiation on human health, especially on fertility and in relation to cancer. Commercially available cellular phones might affect cell function via non-thermal effects. We hypothesized that the plasma membrane might be the target of cell phone radiation. RF-EMW can increase ROS formation by increasing the activity of plasma membrane NADH oxidase. Prolonged exposure to RF-EMW can also cause DNA damage (by prolonged OS), which may accelerates neuronal and spermatozoal cell death and promote neurodegenerative processes as well as promote brain and testicular carcinogenesis. Any tumor promoting effects of RF-EMW might be due to the effect it has on PKC, ODC, intra cellular calcium spikes and stimulation of stress kinase. Stimulation of plasma membrane NADH oxidase might play central role in above mentioned effects.OS and changes in PKC activity might lead to the RF-EMW related infertility observed in numerous studies. Hence, RF-EMW from commercially available cell phones might affect the fertilizing potential of spermatozoa. Therefore, the SAR limit (maximum acceptable exposure limit) should be lowered for cellular phones. However, more studies are necessary to provide definitive evidence against cell phone radiation, which can be provided by in vitro studies combined with computational biomodeling.RF-EMW: Radiofrequency electromagnetic waves; SAR: Specific absorption rate; FDTD: Finite difference time domain; MDA: Malonaldehyde; Hsp: Heat shock protein; PKC: Protein Kinase C; SOD: Superoxide dismustase; GSH-Px: Glutathione peroxidase; CAT: catalase; OS: Oxidative stress; MAPK: Mitogen activated protein kinase; ERK: Extracellular signal regulated kinases; ODC: Ornithine decarboxylase.The authors declare that they have no competing interests.NRD has substantial contribution in designing the article and drafting 50-60% article. KK has substantial contribution in conception and designing article and drafting 20-30% article. AA has critical contribution in drafting article, revision and final approval. All the authors read and approved the final manuscript.


Myodes rufocanus, Microtus fortis and Microtus oeconomus captured in the Baikal area of Buryatia, Russian Federation. Genetic analysis of S- and M-segment sequences of Buryatian hantavirus strains showed that Myodes-associated strains belong to Hokkaido virus (HOKV) type while Microtus-associated strains belong to Vladivostok virus (VLAV) type. On phylogenetic trees Buryatian HOKV strains were clustered together with M. rufocanus- originated strains from Japan, China and Far-East Russia (Primorsky region). Buryatian Microtus- originated strains shared a common recent ancestor with M. fortis- originated VLAV strain from Far-East Russia (Vladivostok area). Our data (i) confirm that M. rufocanus carries a hantavirus which is similar to but distinct from both Puumala virus carried by M. glareolus and Muju virus associated with M. regulus, (ii) confirm that M. fortis is the natural host for VLAV, and (iii) suggest M. oeconomus as an alternative host for VLAV.Hantavirus genome sequences were recovered from tissue samples of  Hantavirus, family Bunyaviridae) are negative-strand RNA viruses with a tripartite genome, each carried by a specific rodent or insectivore host  and also Far-East Russia (Primorsky region) [Myodes- originated sequences showed the host-dependent clustering of PUUV, HOKV and MUJV genetic variants, in good agreement with the idea of co-evolution of hantaviruses with their natural carriers [Similarly to the S segment sequences, the M-sequences recovered from Buryatian  region) . The M scarriers -21.The S segment sequences of two Nesterikha strains were identical; their partial M-sequences differed by one silent substitution only (sequence identity 99.6%). Complete S-sequences of strains Nesterikha503 and Barguzin483 were very close to each other: only nine nucleotide substitutions were found in the coding region (sequence identity 99.3%) and only seven mutations, six substitutions and one deletion, in the 3'-noncoding region that, in this case, could be easily aligned. Seven of nine nucleotide substitutions found in the coding region were silent. Two mutations caused homologous aa substitutions in the deduced sequence of the N protein: Lys96->Arg and Ile326->Val. Partial M segment sequences of these two strains differed by three silent substitutions (sequence identity 99.2%).Two closely related S segment sequences of Buryatian VLAV strains showed the highest level of sequence identity to the prototype wt strain from Vladivostok (Far-East Russia): 87.3–87.4%. The deduced aa sequences of the N protein of two Buryatian VLAV strain were 95.5% identical to the corresponding sequence of Vladivostok strain. The N-sequence identities with closely related KHAV and TOPV were substantially lower: 91.0% and 90.3%, respectively. On the phylogenetic tree constructed for the S segment coding region . Our previous experience tells that, at least in some cases, an addition of one-two "critical" sequences to the dataset could remarkably improve the phylogenetic resolution ,24. SomeM. oeconomus was a bit surprising and thus added a new twist to the already quite intriguing relationships between TOPV, KHAV, and VLAV. KHAV and VLAV, both carried by Microtus voles, do not cluster on the phylogenetic trees with other hantaviruses carried by Microtus  but instead appeared monophyletic with TOPV and the KHAV-VLAV-TOPV trio forms a sister taxon to the Myodes-carried hantaviruses: PUUV, HOKV, and MUJV  in Buryatian population and to evaluate potential threats to human health which might be imposed by these hantaviruses.In this paper, for the first time, we describe HOKV and VLAV strains found in Buryatia, Russian Federation. Although no human cases have been so far attributed to either of these two hantaviruses, further epidemiological studies are needed to estimate a seroprevalence to HOKV and VLAV  declare that they have no competing interests.AngP participated in the screening of rodent samples, performed RT-PCR and sequencing, participated in the genetic analysis and contributed to writing of the manuscript. JL, JN and BP participated in fieldwork and in screening of rodent samples. KN performed genetic analysis of rodents. GM and ÅL participated in the study coordination. AV, HH and OV participated in the study design and coordination, and in drafting the manuscript, the latter two also in fieldwork. AP participated in the study design and coordination, performed (phylo)genetic analyses and contributed to writing of the manuscript. All authors read and approved the final manuscript.


Genetic maps constitute the basis of breeding programs for many agricultural organisms. The creation of these maps is dependent on marker discovery. Melon, among other crops, is still lagging in genomic resources, limiting the ability to discover new markers in a high-throughput fashion. One of the methods used to search for molecular markers is DNA hybridization to microarrays. Microarray hybridization of DNA from different accessions can reveal differences between them--single-feature polymorphisms (SFPs). These SFPs can be used as markers for breeding purposes, or they can be converted to conventional markers by sequencing. This method has been utilized in a few different plants to discover genetic variation, using Affymetrix arrays that exist for only a few organisms. We applied this approach with some modifications for marker discovery in melon.Using a custom-designed oligonucleotide microarray based on a partial EST collection of melon, we discovered 6184 putative SFPs between the parents of our mapping population. Validation by sequencing of 245 SFPs from the two parents showed a sensitivity of around 79%. Most SFPs (81%) contained single-nucleotide polymorphisms. Testing the SFPs on another mapping population of melon confirmed that many of them are conserved.Thousands of new SFPs that can be used for genetic mapping and molecular-assisted breeding in melon were discovered using a custom-designed oligo microarray. A portion of these SFPs are conserved and can be used in different breeding populations. Although improvement of the discovery rate is still needed, this approach is applicable to many agricultural systems with limited genomic resources. Genetic maps based on molecular markers are a key step in marker-assisted selection (MAS) for plant breeding and for gene identification based on positional information . DiscoveIn the last few years, genomic tools have begun to replace the traditional methods of molecular marker discovery. Use of computational tools to search sequence information for molecular markers ,4, or geArabidobsis [Drosophila [One of the technologies that has been used for identification of genetic variation is single-feature polymorphism (SFP) -5. This osophila , and in osophila ,10. To dMelon is among the most important fleshy fruits for fresh consumption. It belongs to the family Cucurbitaceae which includes other important crops, such as watermelon, cucumber and squash. Melon is a diploid species 2n = 24) of African origin with a high level of phenotypic and molecular variation  and a ge of AfricWe chose melon as representative of a large group of vegetables and other crops that are agriculturally important but are still lagging behind in their genomic resources. Although sequencing has become extremely affordable using new technologies, we are still not at a stage at which we can sequence a variety of the same plant again and again. In this paper, we present the ability to use a custom-designed microarray with longer oligos (45-55 mer), based on a partial EST database of melon, to discover thousands of new markers for melon. These markers can be used for genetic mapping, breeding and association studies.Cucumis melo L.: PI414723 (subspecies agrestis) and 'Dulce' (subspecies melo). Two biological replicates from PI414723 and two from 'Dulce' were used. Each biological replicate contained gDNAs pooled from 10 different plants. gDNA samples were labeled and hybridized using standard Agilent procedures for comparative genomic hybridization (CGH) (see Methods).We designed a custom oligo microarray (Agilent) based on the melon ESTs presented in the Cucurbit Genomic database (ICuGI) . The micp-value  and the B statistic, which calculates the log odds that a given SFP will be different between two populations  of 5% was 6184 in 3849 UniGenes. Using the B statistic with B ≥ 1.5, which indicates 82% probability of having differential alleles (SFPs), we found 2503 SFPs in 1771 genes. Due to the fact that the q-value and B statistic rank the SFPs in the same order, and since the B statistic requires prior knowledge of the proportion of SFPs while the moderated-t statistic does not, we used the critical value of moderated-t statistic, i.e., a q-value < 0.05.Predicted SFPs were identified using a linear model for microarray (LIMMA) . The dats Figure . The numArabidopsis and yeast [Since the number of available sequences from 'Dulce' and PI414723 in the database is relatively low, we performed validation by direct sequencing of PCR fragments that were amplified from gDNA of the two parents, PI414723 and 'Dulce'. SFPs were chosen randomly for sequencing (with one limitation applied: the flanking regions of the SFPs were set to 50 bp to be able to identify oligos for PCR). Oligos flanking the SFPs were designed and PCR products were amplified from PI414723 and 'Dulce' gDNA pools: 245 putative SFPs were fully sequenced from both plant cultivars. Identification and validation of a SFP is demonstrated in Figure nd yeast . Analysind yeast . Table 2Arabidopsis [Based on the distribution of SFPs per gene Figure , most ofbidopsis .The ability of SFPs to function as markers in different genetic backgrounds is crucial for their use in MAS, association studies and genetic mapping. As plants host large genetic diversity, we decided to test the use of a set of SFPs across different melon cultivars. We tested this question using parents of another melon mapping population, 'Piel de Sapo' and PI161375 . We perfArabidopsis and yeast [In the present study, we tested the feasibility of identifying genetic polymorphism in melon crops by hybridizing gDNA to custom-designed microarrays based on a partial transcriptome. SFP technology has been previously applied for a variety of genetic applications, such as linkage analysis and association studies, mostly in model organisms such as nd yeast ,32, withnd yeast ,33,34. OThe melon cultivars showed substantial within-population genetic variation  [Arabidopsis studies [With some exception ,32, stud15-20%) , better (15-20%) , and not-20% , be studies . One rea studies . In our  studies , using l studies ). One ca studies . As the  studies .The question of conservation of the SFP markers across melon cultivars was tested using two different mapping populations Figures  and 6. TDeveloping a custom-designed array creates the possibility of using SFP technology in many agricultural systems that do not have an Affymetrix platform and allows much more flexibility in design and use. The low price of the screening and the relatively rapid execution and flexibility make it a useful technology. Although SFP technology is far from perfect, and a reduction in FDRs is needed to make thi approach more efficient, we firmly believe that it can contribute to marker discovery, MAS and WGS approaches in various crop plants. We therefore envision the ability to use WGS (SFP technology and others) as a routine element in the process of MAS in breeding programs as the next challenge in the field of breeding technology.SFPs can be discovered using custom-designed microarrays and partial genomic data. This approach can be used with many agricultural organisms for marker discovery and breeding.m optimization (76.6°C). The Tm, as well as other features of the probes such as cross-hybridization, folding, and low complexity, were summed to total scores with weightings of 0.245, 0.61, 0.025, and 0.12, respectively. The output of this process was 198,500 probes. Probes with a total score lower than 0.3 that included "N" in their sequence were filtered out, leaving a total of 186,600 probes. This design was loaded on the Agilent eArray site for array production https://earray.chem.agilent.com/earray/.A single-feature array was designed based on 16,637 non-redundant cDNA sequences of the Melon Genomics Database from different genotypes and conditions . This seGenomic DNA was extracted using standard procedures  from youAluI and RsaI . The digested DNA was used for labeling and hybridization following Agilent procedures for CGH [gDNA (2.5 μg) was digested with restriction enzymes  for CGH . ScanninProbe signal pre-processing and fitting were performed with the R-package for LIMMA , followi1) A least-square fitting using the following equation:Where the vector of P-D, D-P, D-D, and P-P is the vector of log signal ratios, the matrix of 1, 0, and -1 is the design matrix, and the vector of D2-D1, P1-D1, and P2-D1 is a vector of the estimated coefficients. Next we used these coefficients to estimate three comparisons (contrasts):the inter-variationthe intra-PI variationthe intra-'Dulce' variationp-value was corrected using Benjamini and Hochberg's method [p-value .2) The ordinary t-statistic of each comparison (contrast) for each SFP probe was moderated by shrinking its standard error towards a common value borrowed from the gene ensemble by Bayesian as described in Smyth 2004 . Therefos method  to get ap-value giving the log odds that an SFP is true . The exact formula of B is described in [The B statistic is a Bayesian view of the ribed in .SFPs were selected randomly for validation from the list of putative SFPs. Based on sequence information from the melon database, primers were designed from 50-100 bp of the SFP using Primer3 software . PCRs weSFP: single-feature polymorphism; PPV: positive predictive value; MAS: marker-assisted selection; FDR: false discovery rate; BAC: bacterial artificial chromosome; WGS: whole-genome scanning; gDNA: genomic DNA.AS and RO conceived the idea of exploring melon variation using custom-designed microarrays. RO designed the array and performed all of the statistical analyses. NK helped in integrating the experimental design for melon biology. VP, RB, YB and GT created the biological resources and did some of the wet work for validation of the results. SU was involved in the bioinformatics analysis of the array data and validation of the results. AS and RE contributed to the experimental design and testing of the microarray output. All authors read and approved the final manuscript.List of 245 positive SFPs and their genotyping. The table describes the validation by Sanger sequencing and the difference, where there is one, between the probe and targets. Header descriptions. ProbeName - Probe index. Target - Target resource index as in the melon database v2 http://www.icugi.org/. Sequnce - probe sequence. Position - position of genetic variation within probe sequence. SFP - Either 1 for true positive (TP) or 0 for false positive (FP). NoSNP - number of single-nucleotide polymorphisms (SNPs) in the SFP; 999 indicates many. IndelLength - Indel length if one exists. Pi - PI414723 genotype. Dulce - 'Dulce' genotype. Ref - probe genotype. t - the moderated-t statistic. logFC - log2 of PI-to-'Dulce' signal ratio. adj.P.Val - p-value after multiple-tests correction by Benjamini and Hochberg methodClick here for filemelo L.: PI414723 and 'Dulce' accessionsQualitative estimation of the intra-variation within Cucumis . The document includes description of the experimental design and figure of comparison between two melon accessions, PI414723 and 'Dulce', intra- and inter-variation.Click here for file


Transcriptome variability is due to genetic and environmental causes, much like any other complex phenotype. Ascertaining the transcriptome differences between individuals is an important step to understand how selection and genetic drift may affect gene expression. To that end, extant divergent livestock breeds offer an ideal genetic material.We have analyzed with microarrays five tissues from the endocrine axis  of 16 pigs from both sexes pertaining to four extreme breeds . Using a Bayesian linear model approach, we observed that the largest breed variability corresponded to the male gonads, and was larger than at the remaining tissues, including ovaries. Measurement of sex hormones in peripheral blood at slaughter did not detect any breed-related differences. Not unexpectedly, the gonads were the tissue with the largest number of sex biased genes. There was a strong correlation between sex and breed bias expression, although the most breed biased genes were not the most sex biased genes. A combined analysis of connectivity and differential expression suggested three biological processes as being primarily different between breeds: spermatogenesis, muscle differentiation and several metabolic processes.These results suggest that differences across breeds in gene expression of the male gonads are larger than in other endocrine tissues in the pig. Nevertheless, the strong presence of breed biased genes in the male gonads cannot be explained solely by changes in spermatogenesis nor by differences in the reproductive tract development. It is well known that variability at the transcriptome is in part due to genetic causes, much like any other complex phenotype, e.g., . Thus, tAlthough the differences between tissues and across development stage transcriptomes is a well known fact -4, the vIn order to study the impact of breed differentiation on the pig's transcriptome, we have analyzed the breed and sex differences across different tissues. Among tissues that are of interest, those involved in the different endocrine axes stand out as a promising choice, considering their fundamental biological role and that their transcriptomes have not been widely analyzed. Here we report a detailed microarray analysis of five tissues that make up two main endocrine axes, the HPTA  and HPT  axes, plus fat tissue, in four highly divergent porcine breeds. Both HPTA and HPT axes are highly influential endocrine axes and, we conjectured, must be responsible for at least some of the large phenotypic differences between breeds caused by artificial selection in livestock, e.g., in fat content.The tissues sampled were hypothalamus (HYPO), adenohypophysis (AHYP), thyroid gland (THYG), gonads (GONA) from both sexes, males (GONAM) and females (GONAF), and back fat tissue (FATB). Some of their primary endocrine roles are in Additional file 2), i.e., the variance of the effect divided by the total variance. We and others /SD [(PSg1 - PSg2)|y] where subscripts 1 and 2 refer to males and females, respecively. Similarly, the j-th breed z-score for g-th probeset is defined as zgj = E(PBgj|y)/SD(PBgj|y). A rough measure of how much the expression varies across breeds is computed as the standard deviation of the breed Bayesian z-scores, i.e., for probeset g SDzbreed, g = hclust R function. The distance chosen was one minus the Pearson correlation (r) across variables, this distance is bound between 0 (r = 1) and 2 (r = -1).Bayesian methodology offers several advantages over more traditional least square or maximum likelihood approaches and these will not be discussed here ,31. It satistics . We obtad probesets are either breed or sex biased, and, among them, a subset of nOBS probesets belong to module j. In addition, suppose that nj is the total number of probesets in the module j. The expected number of probesets in the module is simply nEXP = nd × nj/12,000 . This value is corrected when not all nd probesets are among the 12,000 employed for the module analysis. We computed the chi-squared statistics, (nOBS - nEXP)2/nEXP, and the associated P-values. We did this for each module and tissue separately.As genes function in concerted action, which is best described by networks, we studied differential connectivity and checked whether most differentially expressed genes belonged to a specific group of highly co-regulated genes (a module). To do that, we characterized the number of modules using the approach in  employinGene ontologies and GO over representation were analyzed with onto-tools .r2 = 0.97).Hormonal concentrations were measured in duplicate in plasma using commercial testosterone , estradiol  and progesterone EIA kits . All assays were conducted following the manufacturer's protocol. The assays were validated for pig plasma by demonstrating that serial dilutions of plasma were parallel to the displacement curve for the reference standards. Hormone standards spiked with pig plasma produced accurate results of hormone recovery  is also available on request. Contact the author for details.DU: Duroc pig breed; FDR: False Discovery Rate; GO: Gene Ontology; HPT: hypothalamic-pituitary-thyroid axis; HPTA: hypothalamic-pituitary-gonadal axis; IB: Iberian pig breed; GCRMA: GC Robust Multiarray Average; LW: Large White pig breed; QRT-PCR: Real Time Quantitative Reverse Transcription Polymerase Chain reaction; SD: standard deviation; UPGMA: Unweighted Pair-Group Method with Arithmetic Mean; YL: Youli pig breed.All carried out research; MPE conceived research, analyzed data and wrote the manuscript; MLB supervised dissection and co-wrote the manuscript. All authors read and approved the final manuscript.Tissues sampled.Click here for fileComplete list of sex z-scores for every tissue. HYPO: Hypothalamus; AHYP: adenohypophysis; THYG: thyroid gland; GONA: gonads; FATB: backfat. Yellow underlined probesets correspond to the 100 most sex biased in each tissue.Click here for fileOver represented Gene Ontology (GO) categories among the 1700 most sex biased genes in gonads.Click here for fileMeans (SD) of the standard deviations of breed Bayesian z-scores.Click here for fileComplete list of breed z-scores in each tissue. LW: Large White; DU, Duroc; YL, Youli; IB, Iberian; HYPO: Hypothalamus; AHYP: adenohypophysis; THYG: thyroid gland; GONA: gonads; FATB: backfat; SD, standard deviation of breed z-scores for each breed and tissue.Click here for file


Corticotropin-releasing hormone (CRH) is a critical neuropeptide modulating the mammalian stress response. It is involved in many functional activities within various brain regions, among which there is a subset of CRH neurons occupying a considerable proportion of the cortical GABAergic interneurons. Here, we utilized rabies virus-based monosynaptic retrograde tracing system to map the whole-brain afferent presynaptic partners of the CRH neurons in the anterior cingulate cortex (ACC). We find that the ACC CRH neurons integrate information from the cortex, thalamus, hippocampal formation, amygdala, and also several other midbrain and hindbrain nuclei. Furthermore, our results reveal that ACC CRH neurons receive direct inputs from two neuromodulatory systems, the basal forebrain cholinergic neurons and raphe serotoninergic neurons. These findings together expand our knowledge about the connectivity of the cortical GABAergic neurons and also provide a basis for further investigation of the circuit function of cortical CRH neurons. Corticotropin-releasing hormone (CRH) is an important widely expressed neuropeptide with neuroendocrine and neurotransmitter properties, which is essential for brain function . Since iThe recent developed viral tracing system with modified rabies virus, which can map the monosynaptic afferents to a genetically defined neuronal subtype, has been applied to identify the whole-brain presynaptic partners of a specific type of neurons within a complex neural network . Here, wThis study was carried out in accordance with the recommendations of the guidelines issued by the Institutional Animal Care and Use Committees (IACUC) at Wuhan Institute of Physics and Mathematics, the Chinese Academy of Sciences, China. The protocol was approved by IACUC at ShanghaiTech University. Every effort was made to ensure the mice used were treated humanely and any discomfort was kept to a minimum.ad libitum access to rodent food and water freely available in environmentally controlled conditions. The mice used in the study were adult (8–15 weeks) CRH-ires-Cre knock-in mice (Stock No. 012704)  and C57BL/6 mice (N/A) (Shanghai Model Organisms).All mice were housed under a 12/12 day/night cycle at the temperature of 22–25°C, with trans-synaptic retrograde tracing experiments included AAV-CAG-DIO-TVA-GFP , AAV-CAG-DIO-RG , and EnvA-pseudotyped, glycoprotein (RG)-deleted and DsRed-expressing rabies virus  (5.0 × 108 genomic copies per ml), which were packaged and provided by F. Xu . Surgical procedures generally followed previous studies  in PBS. After 2 h of post-fixation in 4% PFA, brain samples were transferred to 30% sucrose (m/v) in 1 × PBS over one night. Then brains prepared with the optimum cutting temperature compound (O.C.T Compound) were sectioned coronally in 50 μm thickness on a freezing microtome (Leica CM1900). One out of every three sections was counterstained with nucleus dye DAPI  and these sections were imaged for all subsequent analyses with an Olympus VS120 microscope. For the quantifications of starter cells and afferent input cells, we divided the boundaries of the subregions, according to the Allen Institute’s reference atlas . FurtherCRH-ires-Cre mice, a genetically engineered mouse line, were used to target CRH neurons specifically. We utilized rabies-based viral strategy to map the whole-brain monosynaptic inputs to the ACC CRH neurons . The rab+ (from the TVA-GFP fusion) and DsRed+ (from rabies virus)  was stereotaxically micro-injected into the bilateral ACC of CRH-ires-Cre mice . The hels virus) , whereasirus) , respectively . Among tThe thalamus has complex functions, generally viewed as a relay station to transfer and integrate sensory signals, including motor signals to the cortical areas, and the modulation of sleep, alertness, learning, and decision-making . The thaOur results demonstrated that the thalamus was the second largest inputs source  of the ACC CRH neurons, showing pervasive and widespread monosynaptic input neurons , 3. AmonPrevious studies have demonstrated that the basal forebrain is a complex nucleus which provides GABAergic, glutamatergic neurons and cholinergic inputs to cortical areas . FunctioThere were a few areas in the midbrain and hindbrain that provided weak but important innervations. The dopamine (DA) neurons originated from either the ventral tegmental area (VTA) or substantia nigra pars compacta (SNc) have been proposed to have complex and multifaceted functions, including modulating appetitive, reward-related behaviors . There atrans-synaptic retrograde tracing experiment, the TH negative neurons maybe GABAergic or glutamatergic.To further provide the new perspectives for the function of the dopaminergic system, we examined the cell type of rabies-labeled neurons in the VTA by immunostaining experiments . The TH,Serotonin , another neuromodulator, has been proposed to have an essential impact on brain functions. Several studies have revealed 5-HT’s involvement in predicting punishment or harm aversion, impulsivity, stress and anxiety, and a wide variety of functions including emotion, sleep, reward, attention, and memory . The rap+TPH2– cells we labeled may be GABAergic neurons. This difference may be quite important for understanding how these two distinct serotonergic systems modulate the limbic system in normal brain and psychiatric disorders. This may give us a more rigorous cue of the circuitry of 5-HT neurons in the brain and help to understand how serotonergic and CRH system interconnected in normal and disease conditions.To further identify the disparity between the monosynaptic inputs to the ACC CRH neurons in the MR and DR, we carried out the immunostaining experiments  and made+ cells in our results, such as the substantia nigra, pars compacta (SNc)  .Compared with conventional retrograde tracing techniques, rabies virus system allows for mapping of monosynaptic inputs to defined neuronal subtypes by combining Cre-loxp system. In this study, we used CRH-ires-Cre mice to target CRH neurons specifically. Through the joint use of CRH-ires-Cre line and rabies virus system, we exclusively distinguished the CRH cell-specific inputs from the general inputs to ACC. As previous studies showed, this method with the combination of these two systems was efficient in labeling the monosynaptic inputs of the cell-type specific neurons . During Pain is a distressing sensory and emotional experience often associated with intense or damaging stimuli alert the individual to withdraw from harmful damage . AnatomiThe function of emotion has been described as to decouple stimulus and response, thus modulating cognition to allow for a suitable adaptation to the environment . EmotionThe raw data supporting the conclusions of this manuscript will be made available by the authors, without undue reservation, to any qualified researcher.This study was carried out in accordance with the recommendations of the guidelines issued by the Institutional Animal Care and Use Committees (IACUC) at Wuhan Institute of Physics and Mathematics, the Chinese Academy of Sciences, China. The protocol was approved by IACUC at ShanghaiTech University. Every effort was made to ensure the mice used were treated humanely and any discomfort was kept to a minimum.JH, JL, WS, YY, and SZ conceptualized the project. SZ and FL performed the majority of experiments. JH, SZ, FL, and CF analyzed the data. JH, SZ, and FL wrote the manuscript with the participation of all other authors.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


N = 101) or in specialized service teams ; and second, to compare IPC associated variables in MH-PCTs vs MH-SSTs.This study has two aims: first, to identify variables associated with interprofessional collaboration (IPC) among a total of 315 Quebec mental health (MH) professionals working in MH primary care teams  was significantly and negatively associated with IPC in SSTs.Improving IPC and making MH teams more successful require the development and implementation of differentiated professional skills in MH-PCTs and MH-SSTs by care managers depending upon the level of care required . Training is also needed for the promotion of interdisciplinary values and improvement of interprofessional knowledge regarding IPC. Interprofessional collaboration (IPC) is defined as a process by which professionals from multiple disciplines share roles and tasks in order to respond in a coherent and integrated way to the needs of patients, their loved ones and the community , 2. IPC Despite abundant evidence for the positive effects of IPC, studies have shown that the uptake of IPC in organizations remains weak; IPC is also inadequately practiced in healthcare teams , 9. Inad. Individual characteristics include demographic variables  and personal attributes  and gender . The mean age of study participants was 43 years for the total sample, 41.7 years in MH-PCTs, and 44.1 years in MH-SSTs. Mean seniority on the teams was 3 years for the total sample, 2.25 years in MH-PCTs, and 3.44 in MH-SSTs. Most participants in both the total sample and the two subsamples were psychosocial professionals (77.2% in MH-PCTs vs. 44.4% in MH-SSTs); while the rest were medical professionals (15.8% in MH-PCTs vs. 43.5% in MH-SSTs) or general professionals (7.9% in MH-PCTs vs. 12.1% in MH-SSTs). IPC had a mean score of 8.022 (SD = 3.751) for PCTs, and 19.931 (SD = 3.747) for SSTs. Participant characteristics are presented in Table Three hundred fifteen (315) MH professionals working in PCTs (SSTs N = 14 partic2), and had acceptable goodness-of-fit. In the MH-SSTs, four variables were independently and positively associated with IPC, of which two related to Interactional characteristics (knowledge integration and team climate), one related to Structural characteristics , and another related to Professional role characteristics . The single variable related to Individual characteristics, age, was significantly and negatively associated with IPC. This model explained 55.0% of total variance (adjusted R2), and had acceptable goodness-of-fit.The multiple linear regression models are presented in Table This study was to our knowledge the first to compare variables associated with IPC in MH-PCTs and MH-SSTs. The findings revealed three variables independently and positively associated with IPC in both PCTs and SSTs ; whereas knowledge sharing correlated with MH-PCTs only, and two other variables  with MH-SSTs only. Therefore, in terms of variables significantly associated with ICP in MH-PCTs and MH-SSTs, there were as many differences as similarities, contrary to our hypothesis. This may be explained by the differences in teams working in primary care as compared with specialized care, described in the introduction, and by the fact that most PCTs were formed following the MH reform with staff from former SSTs. The increased promotion of integrated care among professionals from both PCTs and SSTs aimed at accommodating patients who need service at both levels of care may also explain our results. Moreover, while the proportion of medical professionals was greater among SSTs, and psychosocial professionals more prevalent among MH-PCTs, inter-professional collaboration (IPC) was not associated with type of professional in either MH-PCTs or MH-SSTs.In addition, all four blocks in the conceptual framework were represented for MH-SSTs, whereas only two blocks represented MH-PCTs. No Individual or Structural characteristics variables were associated with IPC in MH-PCTs. This result may be explained by the recent nature of professional transfers to the new multidisciplinary MH-PCTs, and the early stage of team operations in terms of staff acquisition, the transfer of task-related knowledge, skill development as well as team support . The embTwo variables related to Interactional characteristics (knowledge integration and team climate) were associated with IPC in both MH-PCTs and MH-SSTs. A high degree of knowledge integration and positive work environment are particularly important in managing most common MH problems, but also in treating severe and enduring MH problems such as schizophrenia and bipolar disorder. Studies have shown that IPC would be limited without knowledge integration from different disciplines and a positive working environment , 41; whiThe multifocal identification variable under Professional role characteristics was also associated with IPC in both MH-PCTs and MH-SSTs. This underlines the importance of professional identification for improving IPC in MH teams practicing from a multidisciplinary model of care, such as those responsible for expert assessments, case reviews and management of individuals with complex needs. Indeed, professional identification was experienced as a great source of satisfaction and motivation for workers whose professional skills were valued by other team members . MoreoveThe knowledge sharing variable under Interactional characteristics was associated with IPC in MH-PCTs only. This suggests the particular importance of knowledge sharing in the newly formed MH-PCTs dealing with patients mainly affected by common mental disorders and substance use disorders as well as multiple biopsychosocial needs that require sharing of information, skills, and expertise , 82. Con, a climate conducive to the development of good team working relationships [Organizational support has also had positive effects for reducing staff turnover [Organizational support, a Structural characteristics variable, was associated with IPC for MH-SSTs only. Organizational support entails multi-level leadership in overseeing the provision of team resources, adequate space for patient care, clear rules and procedures , 30, a cionships , as wellionships . This suionships , 88. Speturnover  and on jturnover .Regarding Individual characteristics, our study identified a negative association between age and IPC, unlike some previous research (e.g. ), but onFinally, some independent variables identified as significant in other studies were not related to IPC in this research, whether for MH-PCTs or MH-SSTs, which may be partly explained by the study context. However, some variables that were not in collinearity with others, but measured different dimensions, may have been relatively close in meaning . Most astonishing was that type of profession  was not related to IPC in our study. This result may have occurred because teams were too similar in terms of their mix of professionals to be differentially related to IPC, or perhaps due to the way in which each individual team was regrouped in terms of PCTs or SSTs.Despite some important findings, this study had certain limitations. First, the study used a cross-sectional design, which did not permit the formulation of cause and effect inferences about the data. Second, the research did not include a control group, which would have allowed for comparisons. Third, MH-PCTs and MH-SSTs represented a diversified range of teams such as assertive community treatment teams, emergency department teams (SSTs), single access point or psychosocial teams (PCTs), that were not treated as specific team types in the analyses. Treating each team subtype separately may have produced different IPC associated variables. Studying specific teams with a complementary method such as case study analysis may also have brought additional information concerning team processes and dynamics to the results. In addition, there were more SST than PCT teams, and for some, such as emergency department teams, response rates were relatively low. Fourth, some variables identified as key variables in earlier ICP research  were not investigated in the present study , 28. FifThis study was innovative in a number of ways. First, it included a large sample of professionals working in MH-PCTs or MH-SSTs located in four Quebec MH networks. Second, the study tested numerous variables previously identified as strongly related to IPC and organized within a conceptual framework. Moreover, it identified variables associated with IPC in both MH-PCTs and MH-SSTs, comparing IPC associated variables for the two practice settings. The two multivariate regression models identified three independent variables related to Interactional characteristics, and one each for Individual characteristics, Structural characteristics, and Professional role characteristics, respectively. Three independent variables were associated with both MH-PCTs and MH-SSTs, and three were specific to either MH-PCTs or MH-SSTs; as such, they reveal some variation in IPC across levels of care. This suggests the need for managers to promote the development and implementation of differentiated professional skills on teams, depending upon their required level of care provision. Surely, the maintenance of IPC depends upon the contributions of all team members; whereas their effectiveness may be directly influenced by managers themselves. Thus, at the level of specialized services, managers might focus their attention on organizational support without neglecting other variables identified in association with IPC in both MH-PCTs and MH-SSTs . At the primary MH level, managers should focus on the development of knowledge-sharing competencies. Finally, more outreach activities and training of MH professionals are needed to promote interdisciplinary values and skills as well as interprofessional knowledge and IPC. All in all, MH professionals working as members of multidisciplinary teams need preparation and support to know how to work more effectively and collaboratively.


Estrogen receptors promote target gene transcription when they form a dimer, in which two identical (homodimer) or different (heterodimer) proteins are bound to each other. In hormone-dependent cancers, hormone receptor dimerization plays pivotal roles, not only in the pathogenesis or development of the tumors, but also in the development of therapeutic resistance. Protein–protein interactions (PPIs), including dimerization and complex formation, have been also well-known to be required for proteins to exert their functions. The methods which could detect PPIs are genetic engineering  and/or antibody technology  using cultured cells. In addition, visualization of the target proteins in tissues can be performed using antigen–antibody reactions, as in immunohistochemistry. Furthermore, development of microscopic techniques  has made it possible to visualize intracellular and/or intranuclear organelles. We have recently reported the visualization of estrogen receptor dimers in breast cancer tissues by using the in situ proximity ligation assay (PLA). PLA was developed along the lines of antibody technology development, and this assay has made it possible to visualize PPIs in archival tissue specimens. Localization of PPI in organelles has also become possible using super-resolution microscopes exceeding the resolution limit of conventional microscopes. Therefore, in this review, we summarize the methodologies used for studying PPIs in both cells and tissues, and review the recently reported studies on PPIs of hormones. The quaternary structure of proteins, such as their dimerization, is achieved via specific and non-specific non-covalent interactions. Non-specific interactions occur due to hydrophobic effects or van der Waals forces, and specific interactions occur due to electrostatic forces. It is well known that hydrophobicity is the main driving force behind protein folding . Electroco-activators (NCOAs), is required to exert transcriptional activity of ER dimers on target genes [In the nuclear receptor super family, dimerization (homodimerization or heterodimerization) is a crucial process for the activation of nuclear receptors after ligand binding. Activated nuclear receptors then recruit cofactors forming a receptor–cofactor complex. Among all the nuclear receptor super families, the estrogen receptor (ER) is most well studied because it plays an important role in hormone-related growth in hormone-dependent cancers, especially breast and endometrial cancers ,7. ER haet genes . The PPIet genes   has been classically used for detection of PPI in vitro. Co-IP includes using an antigen–antibody complex as a bait and the target interacting protein as prey. Typically, cell lysate is reacted with the bait-specific antibody. Along with the bait protein and any bait-associated proteins, the prey protein (the interaction partner) will be co-precipitated. The bait protein interacts with its specific antibody, which is bound to micro-beads, such as agarose, sepharose, or magnetic beads . Under tIn hormone-dependent cancer research, it is well known that ERα is co-immunoprecipitated with NCOR in MCF-7 cells treated with tamoxifen, but not in untreated or estradiol-treated cells . MohammeResonance energy transfer (RET) technologies such as bioluminescence RET (BRET) and Förster (or Fluorescence) RET (FRET) is also used widely to examine PPI in vitro ,35. FRET2) present in the donor beads to singlet oxygen (1O2) through excitation [1O2 to the thioxene in the acceptor, and radiation (520 to 620 nm) is emitted from the acceptor. Interaction of ER and retinoic acid receptor with their co-activator was screened using the AlphaScreen assay [ER dimerization was reported using both FRET  and BRETcitation . When acen assay . These fSeveral protein detection methods are available for in vitro, ex vivo, and in vivo analysis. In 1960, radio immunoassay (RIA) was established for measurement of insulin in plasma by Yalow, a Nobel Prize laureate . Enzyme Immunohistochemistry is a classical microscopy-based technique for visualizing the target protein in tissues and cells using a specific antibody. Immunohistochemistry was invented in 1941 by Coons and co-investigators  and is aImmunohistochemistry is currently widely used at the global level. It is possible to identify the localization of a target protein in normal tissues and disease in routinely processed tissue specimens by immunohistochemistry. Steroid hormone receptors can also be detected by immunohistochemistry in several hormone-dependent cancers . ImmunocIn addition, protein localization at the organelle level can be detected using transmission and immunoelectron microscopes . In immuKocanova et al.  demonstrAdvanced techniques based on immunohistochemistry have been developed, and in situ proximity ligation assay (PLA) was developed to visualize PPI of two proteins in tissue samples ,66. ThisWe recently demonstrated that ER dimerization was induced by estrogen treatment in breast cancer cell lines . ERα homIn addition, we also reported the visualization of heterodimers of ERβ variants in breast carcinoma cell lines ,68. In aInteraction of androgen receptor with JunD in prostate cancer LNCaP cells was identified by PLA, and this interaction was inhibited by a selected lead chemical . We receOptical microscopy has contributed immensely to the elucidation of the microstructure of cells, but the resolution of an optical microscope is limited because of its diffraction limit, explained by the Abbe’s law ,87. Intrco-investigators [Gonadotrophic hormones such as follicle-stimulating hormone (FSH) and luteinizing hormone (LH) derived from the pituitary gland, bind to their corresponding receptors in ovarian granulosa (FSH-R) and theca (LH-R) cells, respectively. Jones and tigators  demonstrtigators . Dimeriztigators . LH-R istigators . In breatigators ,97. HERstigators ,97. TherIn our previous study, immunoreactivity of CEACAM6 in breast carcinoma tissues was not associated with the efficacy of HER2 inhibitor, trastuzumab, in the breast cancer patients examined . HoweverInhibitors that directly target estrogen receptor/co-activator binding (co-activator binding inhibitors: CBI) have been developed ,101,102.Since scanning tunneling microscopy and atomic force microscopy (AFM) were developed in the 1980s, a wide variety of scanning probe microscopy (SPM) techniques have been developed based on their principles . The priIn recent years, PPI has been considered one of the important concepts in drug discovery, and PPI itself is regarded as a drug target. PPI is a key mechanism in understanding steroid hormone signals, and is an indispensable part of hormone-dependent cancer research. Furthermore, evaluation of PPI is thought to provide useful information, not only as a diagnostic tool for disease, but also as a predictive marker for the efficacy of the molecular-targeted agents.


The MG-RAST API provides search capabilities and delivers organism and function data as well as raw or annotated sequence data via the web interface and its RESTful API. For casual users, however, RESTful APIs are hard to learn and work with.We created the graphical MG-RAST API explorer to help researchers more easily build and export API queries; understand the data abstractions and indices available in MG-RAST; and use the results presented in-browser for exploration, development, and debugging.The API explorer lowers the barrier to entry for occasional or first-time MG-RAST API users. Environmental DNA sequence analysis  is gaining popularity and is frequently used by researchers who are not specialists in genomics or bioinformatics . With exAlthough significant advances have been made  ) 4]) thaHosted services such as MG-RAST , the EBIIn addition, studying larger quantities of datasets is often done best via custom scripts or command line tools. APIs play an essential role by allowing automation. We also note that APIs render the practice of extracting data from web pages (“screen scraping”) obsolete.In recent years RESTful APIs  have becFor example, MG-RAST is designed around a RESTful API . This prUnfortunately, although RESTful APIs offer more flexibility to access computation and data, they are notoriously difficult to learn . Two facThe API explorer is implemented as light-weight JavaScript and HTML overlay on top of the MG-RAST API  and the https://github.com/MG-RAST under a BSD-style license.Like all of MG-RAST, the API explorer is available as open source software on GitHub at https://explorer.mg-rast.org/). It has one page for search and one for all other API functionality. The explorer allows querying capabilities and displays the results of an API query in-browser. Additionally, it constructs working command line invocations that can be copied and pasted.To assist researchers in learning the syntax of the MG-RAST RESTful API, the MG-RAST team has developed an API explorer , and cuExample 2. Query composition. The next example involves a simple query requesting a list of aquatic datasets collected in Chicago. Figure Example 3: Dataset properties. Searches for datasets can also include technical properties such as dataset size or type. This example retrieves all shotgun metagenomic datasets using Illumina technology larger than 1 gigabase pair. The resulting URL and command line are given in Fig. Corynebacterium glutamicum), taxonomic names from the NCBI taxonomy, or protein function labels from the included protein function databases. See [In addition to storing metadata about the datasets, MG-RAST stores the results of the sequence analysis, indexed to allow querying by organism name .The MG-RAST team hopes that the drop-down menus and in-browser troubleshooting environment provided by the MG-RAST RESTful API explorer will help researchers make better use of the metagenomic data and computation already completed and curated at MG-RAST Fig. . The MG-The API explorer is available as part of MG-RAST.Project name: MG-RASTProject home page:https://github.com/MG-RASTOperating system(s): LinuxProgramming language: Perl, Python, Go-Lang, HTML5Other requirements: ElasticSearch, Cassandra, SOLR, MongoDB, SHOCK, AWELicense: BSD type licenseAny restrictions to use by non-academics: none.


In recent years, the demand for natural herbal products (NHP) has increased; however, the quality of these products is difficult to confirm due to the lack of a comprehensive quality control system. Traditional methods are not effective in detecting processed ingredients. DNA barcoding is an established technique that has been used for more than 10 years. This technique uses short standard sequences  to identify species. While a complete DNA barcode is difficult to obtain from NHP due to DNA degradation, mini-barcoding is a complementary tool to identify species in NHP. DNA mini-barcoding uses smaller DNA segments for polymerase chain reaction amplification and can be applied to identify species rapidly. The present review summarizes the development and application of DNA mini-barcodes over recent years and discusses the limitations of this technique. This review also compares mini-barcoding and meta-barcoding, a technique using universal polymerase chain reaction primers to simultaneously amplify multiple DNA barcodes and identify many species in a single environmental sample. Additionally, other detection methods that can be combined with mini-barcodes, such as nucleotide signatures, high-resolution DNA melting analysis, and gold nanoparticles, are discussed. DNA mini-barcoding can fill the gaps left by other methods in the field of herbal molecular identification. Natural herbal products (NHP) use herbs as raw materials and include herbal medicines, dietary supplements, and herbal extracts. In recent years, worldwide demand for NHP has increased, making NHP adulteration and counterfeiting a global problem . ApproxiThe currently available NHP chemical detection methods are not sufficient . These mmatK), photosystem II protein D1-tRNA-His, ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit (rbcL), internal transcribed spacer (ITS), internal transcribed spacer 2 (ITS2), and cytochrome c oxidase I (CO1)  . DNA bar I (CO1) . DNA bar I (CO1) .DNA barcoding has been successful in the efficient and inexpensive identification of herbal materials; however, it is not suitable for identifying processed NHP . DNA degDNA mini-barcoding, using a smaller length of DNA compared with traditional barcoding, may help overcome the difficulties associated with DNA barcoding. . Mini-baThe present review emphasizes the utility of DNA mini-barcoding, summarizes key technologies utilized in this field, and discusses deficiencies associated with this technique. Furthermore, this review outlines technologies that could be developed in combination with mini-barcoding to improve NHP identification.NHP often undergoes complex processing, including pulverization, extraction, leaching, purification, concentration, drying, and granulation. For example, the production of Shuanghuanglian particles involves boiling Lonicerae japonicae Flos for 1.5 h, batch filtering, and concentration . As thistrnL) in chloroplasts, termed P6 loop (10–143 bp), could be amplified from processed food and permafrost samples  Liuwei Dihuang Wan, amplified the  the DNA . Lo et a the DNA . Song an the DNA . Table 1In the cells of medicinal plants, secondary metabolites including fibers, proteins, polysaccharides , parenchLow-quality DNA is difficult to amplify and sequence; thus, DNA quality greatly influences the identification efficiency of the mini-barcoding method . Chen etExtraction of high-quality DNA from NHP is the initial step and is critical to successfully apply molecular diagnostics . ConventContamination during DNA extraction is also a concern. Low-quality DNA can be easily contaminated by fungal, bacterial, or other herbal materials, which can interfere with PCR amplification .rbcL > 500).Typical PCR amplification of DNA barcodes is based on universal primer sets . HoweverrbcL region  should be the initial standard to screen a marker for a mini-barcode. A reliable database of standard sequences should be established. These sequences could be collected from the GenBank, European Molecular Biology Laboratory, or DNA Data Bank of Japan, which have been published or uploaded by authorities. Subsequently, candidate sequences may be aligned to identify conserved and specific regions. Each mini-barcode should be proven to be unique using the Basic Local Alignment Search Tool (BLAST) analysis in standard databases with 100% identity. In addition, specific primer pairs for mini-barcode amplification should be designed for later PCR amplification. For instance, Hofreiter et al. designed primer pairs for a 157 bp amplicon in the L region . Lo et agranules .Multiple sequencings in both the forward and reverse directions can be aligned and assembled into consensus contigs using software , and lowAlthough DNA mini-barcodes can help identify processed products, this technique is limited by its length constraint.The purpose of the development of DNA mini-barcodes is not to achieve universal application for most species, but rather to identify specific targets for medicinal plants. Every PCR amplification and taxonomic result must be tested and applied. DNA mini-barcoding is unable to identify unknown adulterants or contamination, especially in complex NHP that occasionally contain >10 species. Moreover, sequences of different lengths are chosen as a mini-barcode in terms of different cases. As a result, they could not develop into standard biomarkers when dealing with all cases . In addiEphedrae herbs  as the entire ITS2 sequence fails to identify these species  as biomarkers to analyze nine commercial Liuwei Dihuang Wan specimens in three batches using high-throughput sequencing (HTS)  . Differedetected .Certain aspects of DNA mini-barcoding are similar to meta-barcoding. Both require shorter sequencing regions than regular barcodes (200–300 bp), and these shorter regions are amplified and sequenced. Nevertheless, differences exist between the technologies. Several preparation steps, including sequence screening and primer design are critical in mini-barcoding. The purpose of meta-barcoding is to simultaneously obtain sequences from as many species as possible. Consequently, universal primers applicable to multiple species, rather than species-specific primers, are warranted. Moreover, DNA mini-barcoding is frequently used to identify one or several related species in CPMs, while meta-barcoding is able to simultaneously identify a wide range of species or identify bulk biodiversity samples and preparations.In meta-barcoding studies, a large quantity of sequence data must be obtained and analyzed, rendering the Sanger sequencing technique unsuitable. The next-generation Illumina sequencing system solves this problem inexpensively. However, the Illumina system is also characterized by limitations. NGS is generally accurate, especially in genome studies, while it is not the case when dealing with meta-barcoding studies of CPMs. This is because the heavily degraded DNA is unsuitable to establish a sequencing library. More PCR amplification cycles are thus warranted to increase the quality of DNA for sequencing, and the quality of sequencing reads may be substantially lower in subsequent cycles. For instance, Dohm et al. demonstrated that miscalls were distributed in guanine cytosine-rich (GC-rich) regions, and base-specific miscalls C to G or A to C were more often observed in later PCR cycles . MoreoveIn conclusion, NGS can yield large amounts of sequencing data and efficiently identify multiple species. However, sequencing errors during NGS may interfere with accurate species identification. Thus, it is necessary to align sequences along every base, allowing more accurate and reliable alignment results to be obtained. Furthermore, the analysis of unique regions of each broken barcode or mini-barcode is important. The combination of mini-barcoding and meta-barcoding has significant potential for broadening the application of barcoding for NHP identification.It is occasionally necessary to rapidly detect specific ingredients in herbal medicines, and developments in DNA mini-barcoding have increased the efficiency and utility of this technique. Nucleotide signatures developed from DNA barcodes are currently considered the standard for molecular detection of certain herbal materials, avoiding sequencing. Bar-HRM analysis is based on different melting temperatures of single nucleotides. Gold nanoparticle sensors are based on a colorimetric reaction to detect single-nucleotide polymorphisms (SNP) using electrostatic adsorption. These developments are representative of the widely used technologies to detect certain species.A nucleotide signature is an extremely short DNA sequence (20–50 bp), which is unique to a species and highly conserved, with 100% sequence identity through BLAST analysis. Mini-barcoding is appropriate to detect these sequences. Nucleotide signatures were initially proposed by Han’s group for identifying the TCMs Asian ginseng and American ginseng . NucleotDNA mini-barcodes are shorter than a typical DNA barcode. DNA barcodes are not species-specific due to close relationships or frequent variances within a genus. To amplify a target mini-barcode, specific primer pairs replace universal primer pairs and narrow the range of species identification. Nucleotide signatures occasionally match specific primer sets and show amplification specificity. Moreover, they can also enhance the success rate and efficiency of PCR.Panax quinquefolium and Panax ginseng in CPMs. Adulteration of these species can be distinguished using two SNP sites based on the mini-barcode sequences. In addition, the relative heights of the peaks in the sequencing results can be used to deduce the adulteration ratio (Angelicae sinensis radix. The nucleotide signature could be detected in the sequencing results of 19 CPMs labeled with A. sinensis radix. Different adulterants, such as Ligusticum sinense, Notopterygium incisum, Angelica decursiva, and Angelica gigas, were detected in different batches (Lonicera extracts and CPMs. The results revealed the samples contained only 17 and 22% of authentic material, respectively. Two adulterants, namely Eucommiae Folium and Lonicerae Flos, were both detected in 7% of NHP in this study (Nucleotide signatures have been used to authenticate several CPMs. For example, it is difficult to distinguish on ratio . Based oon ratio . Wang et batches . A nucleis study .Panax notoginseng, Cistanche deserticola (Ginkgo biloba (Thus far, nucleotide signatures for erticola , Eucommierticola , and Gino biloba  have beeMelting curve analysis, first introduced in the 1990s , differsSenna alexandrina Mill in crude drugs. They found that Senna species could not be distinguished based on the 700–800-bp PCR amplicons of the ITS region. However, species-specific SNP sites and insertion–deletion mutations were detected at a total of 143 sites in the ITS region of Senna italica subsp. italica and S. alexandrina, which allowed successful simultaneous discrimination (Bar-HRM analysis combines DNA barcodes with HRM analysis and can detect genetic variants in DNA sequences. Bar-HRM analysis allows for high-throughput, rapid, and simultaneous identification of species, and it is suitable for NHP frequently adulterated with related species and genetic variants. Sequences with different variants can be clearly distinguished according to their different melting curves. Bar-HRM has been widely applied to analyze many raw herbal materials and adulterants . Mishra mination .Phyllanthus products were discriminated using Bar-HRM analysis. Two loci from rbcL and trnL were selected to generate suitable primer sets for HRM analysis specificity. Subsequently, 88- and 99-bp amplicons were produced under annealing temperatures of 50–60°C, which were the expected PCR products for the rbcL and trnL loci, respectively (In addition to Bar-HRM, research on herbal products with mini-Bar-HRM analysis has emerged in recent years. In one study, five medicinal ectively .matK regions, coupled with HRM analysis, to authenticate Hypericum perforatum and Hypericum androsaemum in herbal infusions. Species-specific primers were developed for barcoding regions identified through PCR and real-time PCR. One primer was used to produce barcode amplicons for the regular sequencing of Hypericum. Another primer was designed to enable unequivocal identification through HRM analysis. When coupled with HRM, mini-barcoding of matK could differentiate both species. Moreover, mini-barcoding of ITS1 could identify intra-species variability, which was confirmed by HRM analysis (Costa et al. developed mini-barcoding for the ITS1 and analysis .HRM analysis has been used in many studies, indicating its value for sensitive, rapid, and cost-effective identification in combination with DNA barcoding or mini-barcoding. This technique is interchangeable with conventional post-PCR genotyping methods and can be combined with SNP sites or single sequence repeat markers to identify NHP.DNA-gold nanoparticle sensors have been used to rapidly detect diseases, mycotoxins, and pathogenic microorganisms . Gold naThis method involves five items: magnetic particles, AuNPs, a single-stranded probe connected to magnetic particles, a single-stranded probe connected to AuNPs, and a single-stranded target barcode sequence. Magnetic particles coated with a single-stranded probe recognize one end of the target barcode DNA, and AuNPs conjugated to a single-stranded probe combine with the other end of the target barcode DNA. This brings the magnetic particles and AuNPs together through the formation of double-stranded DNA, resulting in a magnetic particle-probe-target barcode-probe-AuNP sandwich structure . The sanAuNPs can be combined with various bioactive molecules, such as biocompatible oligonucleotides . In addiFritillaria thunbergii and Fritillaria cirrhosa (Several molecular detection methods based on nanoparticles have been established to evaluate plant materials. Huang et al. developed an amplified sensor to detect DNA in genetically modified organisms using AuNPs and an electrochemical biosensor, demonstrating good sensitivity . Lei et cirrhosa .Mini-barcoding combined with AuNP sensors could be designed using a similar framework to that of biosensors used to study genetically modified organisms and detect related species. Owing to the high sensitivity of this superior signal amplification platform, mini-barcoding in combination with AuNPs could solve several problems, including unavoidable degradation in NHP, low concentrations of extracted DNA, and low PCR amplification efficiency.The experimental procedures of three key technologies are briefly illustrated in Recent media reports and scientific studies highlight how widespread adulterations and ingredient substitution in NHP have become and underscore the threat to consumer safety.DNA mini-barcoding can overcome NHP identification difficulties caused by DNA degradation after extensive processing. While this method will never supplant typical DNA barcoding in identifying specimens, it can significantly improve the efficiency and accuracy of sample analysis by increasing the PCR amplification rate. New rapid detection methods based on mini-barcoding are expected to be established, particularly for processed products.In general, mini-barcodes are species-specific, allowing their utility in analyzing complex samples. DNA mini-barcodes represent an improved approach to carefully identify plant products and protect public health. This significant tool should be further developed and applied to the quality control of NHP.This review was drafted by ZG and revised by JH and YL. Other data or references were collected by XiW and XuW. All authors have read and approved the final manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.