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1.
Reprod Domest Anim ; 59(4): e14566, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38627959

ABSTRACT

Early pregnancy loss is a primary cause of low reproductive rates in dairy cows, posing severe economic losses to dairy farming. The accurate diagnosis of dairy cows with early pregnancy loss allows for oestrus synchronization, shortening day open, and increasing the overall conception rate of the herd. Several techniques are available for detecting early pregnancy loss in dairy cows, including rectal ultrasound, circulating blood progesterone, and pregnancy-associated glycoproteins (PAGs). Yet, there is a need to improve on existing techniques and develop novel strategies to identify cows with early pregnancy loss accurately. This manuscript reviews the applications of rectal ultrasound, circulating blood progesterone concentration, and PAGs in the diagnosis of pregnancy loss in dairy cows. The manuscript also discusses the recent progress of new technologies, including colour Doppler ultrasound (CDUS), interferon tau-induced genes (ISGs), and exosomal miRNA in diagnosing pregnancy loss in dairy cows. This study will provide an option for producers to re-breed cows with pregnancy loss, thereby reducing the calving interval and economic costs. Meanwhile, this manuscript might also act as a reference for exploring more economical and precise diagnostic technologies for early pregnancy loss in dairy cows.


Subject(s)
Cattle Diseases , Progesterone , Pregnancy , Female , Cattle , Animals , Abortion, Veterinary/diagnosis , Reproduction , Fertilization , Glycoproteins , Insemination, Artificial/veterinary , Cattle Diseases/diagnosis
2.
Chin J Integr Med ; 30(5): 468-479, 2024 May.
Article in English | MEDLINE | ID: mdl-38329654

ABSTRACT

Activation of the NOD-like receptor protein 3 (NLRP3) inflammasome signaling pathway is an important mechanism underlying myocardial pyroptosis and plays an important role in inflammatory damage to myocardial tissue in patients with cardiovascular diseases (CVDs), such as diabetic cardiomyopathy, ischemia/reperfusion injury, myocardial infarction, heart failure and hypertension. Noncoding RNAs (ncRNAs) are important regulatory factors. Many Chinese medicine (CM) compounds, including their effective components, can regulate pyroptosis and exert myocardium-protecting effects. The mechanisms underlying this protection include inhibition of inflammasome protein expression, Toll-like receptor 4-NF-κB signal pathway activation, oxidative stress, endoplasmic reticulum stress (ERS), and mixed lineage kinase 3 expression and the regulation of silent information regulator 1. The NLRP3 protein is an important regulatory target for CVD prevention and treatment with CM. Exploring the effects of the interventions mediated by CM and the related mechanisms provides new ideas and perspectives for CVD prevention and treatment.


Subject(s)
Cardiovascular Diseases , Inflammasomes , Medicine, Chinese Traditional , Pyroptosis , Animals , Humans , Cardiovascular Diseases/metabolism , Inflammasomes/metabolism , NLR Family, Pyrin Domain-Containing 3 Protein/metabolism , Pyroptosis/drug effects , Signal Transduction/drug effects
3.
Pest Manag Sci ; 80(3): 996-1007, 2024 Mar.
Article in English | MEDLINE | ID: mdl-37830147

ABSTRACT

BACKGROUND: The sugarcane borer Diatraea saccharalis (Lepidoptera) is a key pest on sugarcane and other grasses in the Americas. Biological control as well as insecticide treatments are used for pest management, but economic losses are still significant. The use of female sex pheromones for mating disruption or mass trapping in pest management could be established for this species, provided that economical production of pheromone is available. RESULTS: Combining in vivo labelling studies, differential expression analysis of transcriptome data and functional characterisation of insect genes in a yeast expression system, we reveal the biosynthetic pathway and identify the desaturase and reductase enzymes involved in the biosynthesis of the main pheromone component (9Z,11E)-hexadecadienal, and minor components hexadecanal, (9Z)-hexadecenal and (11Z)-hexadecenal. We next demonstrate heterologous production of the corresponding alcohols of the pheromone components, by expressing multiple steps of the biosynthetic pathway in yeast. CONCLUSION: Elucidation of the genetic basis of sex pheromone biosynthesis in D. saccharalis, and heterologous expression in yeast, paves the way for biotechnological production of the pheromone compounds needed for pheromone-based pest management of this species. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.


Subject(s)
Moths , Saccharum , Sex Attractants , Female , Animals , Sex Attractants/chemistry , Saccharomyces cerevisiae , Moths/genetics , Pheromones
4.
Minim Invasive Ther Allied Technol ; 31(8): 1123-1130, 2022 Dec.
Article in English | MEDLINE | ID: mdl-36260704

ABSTRACT

PURPOSE: Approximately 20% of patients with lung nodules (LNs) have multiple LNs (MLNs). This meta-analysis was performed to assess the safety and efficacy of computed tomography (CT)-guided localization of MLNs in comparison with those of single LN (SLN) localization. MATERIAL AND METHODS: The PubMed, Embase, and Cochrane Library were searched to collect relevant articles published till February 2022. The meta-analysis was performed using the RevMan v5.3. RESULTS: In total, seven studies met the inclusion criteria for this meta-analysis. No significant difference was observed between patients with MLNs and SLN in terms of pooled successful localization rate based on LNs (p = 0.64) and patients (p = 0.06). The pooled duration of localization was significantly shorter and the pooled pneumothorax and lung hemorrhage rates were significantly lower in the SLN group than in the MLNs group (p < 0.00001 for all). The pooled duration of hospital stay was comparable between the MLNs and SLN groups (p = 0.96). Significant heterogeneity was observed in the endpoints of duration of localization (I2 = 75%) and pneumothorax (I2 = 53%). CONCLUSIONS: CT-guided simultaneous MLN localization is clinically safe and effective, despite requiring a longer procedural time and having higher incidence of pneumothorax and lung hemorrhage than SLN localization.


Subject(s)
Multiple Pulmonary Nodules , Tomography, X-Ray Computed , Humans , Multiple Pulmonary Nodules/diagnostic imaging , Multiple Pulmonary Nodules/surgery , Pneumothorax/epidemiology , Pneumothorax/etiology , Tomography, X-Ray Computed/adverse effects , Tomography, X-Ray Computed/methods
5.
Metab Eng ; 72: 391-402, 2022 07.
Article in English | MEDLINE | ID: mdl-35598886

ABSTRACT

Biologically produced wax esters can fulfil different industrial purposes. These functionalities almost drove the sperm whale to extinction from hunting. After the ban on hunting, there is a niche in the global market for biolubricants with properties similar to spermaceti. Wax esters can also serve as a mechanism for producing insect sex pheromone fatty alcohols. Pheromone-based mating disruption strategies are in high demand to replace the toxic pesticides in agriculture and manage insect plagues threatening our food and fiber reserves. In this study we set out to investigate the possibilities of in planta assembly of wax esters, for specific applications, through transient expression of various mix-and-match combinations of genes in Nicotiana benthamiana leaves. Our synthetic biology designs were outlined in order to pivot plant lipid metabolism into producing wax esters with targeted fatty acyl and fatty alcohols moieties. Through this approach we managed to obtain industrially important spermaceti-like wax esters enriched in medium-chain fatty acyl and/or fatty alcohol moieties of wax esters. Via employment of plant codon-optimized moth acyl-CoA desaturases we also managed to capture unusual, unsaturated fatty alcohol and fatty acyl moieties, structurally similar to moth pheromone compounds, in plant-accumulated wax esters. Comparison between outcomes of different experimental designs identified targets for stable transformation to accumulate specialized wax esters and helped us to recognize possible bottlenecks of such accumulation.


Subject(s)
Esters , Fatty Alcohols , Esters/metabolism , Fatty Alcohols/metabolism , Pheromones/metabolism , Plant Leaves/metabolism , Nicotiana/genetics , Nicotiana/metabolism , Waxes/metabolism
6.
BMC Biol ; 20(1): 80, 2022 03 31.
Article in English | MEDLINE | ID: mdl-35361182

ABSTRACT

BACKGROUND: Using genetically modified plants as natural dispensers of insect pheromones may eventually become part of a novel strategy for integrated pest management. RESULTS: In the present study, we first characterized essential functional genes for sex pheromone biosynthesis in the rice stem borer Chilo suppressalis (Walker) by heterologous expression in Saccharomyces cerevisiae and Nicotiana benthamiana, including two desaturase genes CsupYPAQ and CsupKPSE and a reductase gene CsupFAR2. Subsequently, we co-expressed CsupYPAQ and CsupFAR2 together with the previously characterized moth desaturase Atr∆11 in N. benthamiana. This resulted in the production of (Z)-11-hexadecenol together with (Z)-11-hexadecenal, the major pheromone component of C. suppressalis. Both compounds were collected from the transformed N. benthamiana headspace volatiles using solid-phase microextraction. We finally added the expression of a yeast acetyltransferase gene ATF1 and could then confirm also (Z)-11-hexadecenyl acetate release from the plant. CONCLUSIONS: Our results pave the way for stable transformation of plants to be used as biological pheromone sources in different pest control strategies.


Subject(s)
Moths , Sex Attractants , Animals , Moths/genetics , Pheromones/metabolism , Nicotiana/genetics
7.
J Ultrasound Med ; 41(3): 617-626, 2022 Mar.
Article in English | MEDLINE | ID: mdl-33938029

ABSTRACT

OBJECTIVES: This study aimed to determine ultrasonic image characteristics that enable differentiation between cholesterol and adenomatous polyps and to assess the diagnostic efficacy of combining conventional ultrasound (CUS) with contrast-enhanced ultrasound (CEUS). METHODS: Eighty-nine patients with gallbladder polyps of 1-2 cm in diameter were enrolled and examined by CUS and CEUS before cholecystectomy. The appearances on CUS and CEUS were recorded and analyzed. The receiver operating characteristic (ROC) curve was used to calculate the optimal size threshold for distinguishing cholesterol from adenomatous polyps. A logistic regression analysis was performed to identify diagnostic variables. ROC analysis was performed to evaluate the diagnostic efficacy of the size, the independent variables, and the combined factors. RESULTS: There were differences in size, number, vascularity on CUS and intralesional vascular shape, wash-out, and area under the curve on CEUS between the two groups (P < .05). ROC analysis indicated that a maximum diameter of 1.45 cm was the optimal threshold for the prediction of adenomatous polyps. The logistic regression analysis proved that the single polyp, presence of vascularity, and intralesional linear vessels were associated with adenomatous polyps (P < .05). ROC analysis showed that the area under the ROC curve, sensitivity, and specificity for the combination of the three independent variables were 0.858, 87.3%, and 67.6%. The number combined with intralesional vascular shape had the highest diagnostic sensitivity of 91.2%. CONCLUSIONS: The combination of CUS and CEUS demonstrated great significance in the differential diagnosis of cholesterol and adenomatous polyps.


Subject(s)
Adenomatous Polyps , Gallbladder Neoplasms , Polyps , Adenomatous Polyps/diagnostic imaging , Cholesterol , Contrast Media , Diagnosis, Differential , Gallbladder Neoplasms/diagnosis , Humans , Polyps/diagnostic imaging , Ultrasonography
8.
Pest Manag Sci ; 78(3): 1048-1059, 2022 Mar.
Article in English | MEDLINE | ID: mdl-34773383

ABSTRACT

BACKGROUND: (Z,E)-9,12-tetradecadienyl acetate (ZETA, Z9,E12-14:OAc) is a major sex pheromone component for many stored-product moth species. This pheromone is used worldwide for mating disruption, detection, monitoring, and mass trapping in raw and processed food storage facilities. In this study, we demonstrate the biological production of ZETA pheromone by engineered yeast Saccharomyces cerevisiae. RESULTS: We mined the pheromone gland transcriptome data of the almond moth, Ephestia (Cadra) cautella (Walker), to trace a novel E12 fatty acyl desaturase and expressed candidates heterologously in yeast and Sf9 systems. Furthermore, we demonstrated a tailor-made ZETA pheromone bioproduction in yeast through metabolic engineering using this E12 desaturase, in combination with three genes from various sources coding for a Z9 desaturase, a fatty acyl reductase, and an acetyltransferase, respectively. Electrophysiological assays (gas chromatography coupled to an electroantennographic detector) proved that the transgenic yeast-produced ZETA pheromone component elicits distinct antennal responses. CONCLUSION: The reconstructed biosynthetic pathway in yeast efficiently produces ZETA pheromone, leaves an undetectable level of biosynthetic intermediates, and paves the way for the economically competitive high-demand ZETA pheromone's bioproduction technology for high-value storage pest control.


Subject(s)
Moths , Sex Attractants , Animals , Moths/genetics , Pheromones , Saccharomyces cerevisiae/genetics , Spodoptera
9.
J Chem Ecol ; 47(12): 950-967, 2021 Dec.
Article in English | MEDLINE | ID: mdl-34762210

ABSTRACT

Synthetic pheromones have been used for pest control over several decades. The conventional synthesis of di-unsaturated pheromone compounds is usually complex and costly. Camelina (Camelina sativa) has emerged as an ideal, non-food biotech oilseed platform for production of oils with modified fatty acid compositions. We used Camelina as a plant factory to produce mono- and di-unsaturated C12 chain length moth sex pheromone precursors, (E)-9-dodecenoic acid and (E,E)-8,10-dodecadienoic acid, by introducing a fatty acyl-ACP thioesterase FatB gene UcTE from California bay laurel (Umbellularia californica) and a bifunctional ∆9 desaturase gene Cpo_CPRQ from the codling moth, Cydia pomonella. Different transgene combinations were investigated for increasing pheromone precursor yield. The most productive Camelina line was engineered with a vector that contained one copy of UcTE and the viral suppressor protein encoding P19 transgenes and three copies of Cpo_CPRQ transgene. The T2 generation of this line produced 9.4% of (E)-9-dodecenoic acid and 5.5% of (E,E)-8,10-dodecadienoic acid of the total fatty acids, and seeds were selected to advance top-performing lines to homozygosity. In the T4 generation, production levels of (E)-9-dodecenoic acid and (E,E)-8,10-dodecadienoic acid remained stable. The diene acid together with other seed fatty acids were converted into corresponding alcohols, and the bioactivity of the plant-derived codlemone was confirmed by GC-EAD and a flight tunnel assay. Trapping in orchards and home gardens confirmed significant and specific attraction of C. pomonella males to the plant-derived codlemone.


Subject(s)
Brassicaceae/chemistry , Dodecanol/analogs & derivatives , Metabolic Engineering , Moths/drug effects , Sex Attractants/pharmacology , Animals , Dodecanol/chemistry , Dodecanol/metabolism , Sex Attractants/chemistry
10.
Zhongguo Zhong Yao Za Zhi ; 46(13): 3270-3287, 2021 Jul.
Article in Chinese | MEDLINE | ID: mdl-34396746

ABSTRACT

The multi-component pharmacokinetic study of Chinese herbal extracts elaborates the in vivo processes,including absorption,distribution,metabolism,and excretion,of multiple bioactive components,which is of significance in revealing pharmacodynamic material basis of Chinese herbal medicine. In recent years,with the innovation in ideas,and development of techniques and methods on traditional Chinese medicine( TCM) research,the pharmacokinetic studies of Chinese herbal extracts were extensively performed,and notable progress has been made. This paper reviewed the advancement of multi-component pharmacokinetics of Chinese herbal extracts in recent five years from analysis technology of biological sample,the pharmacokinetic characteristics of Chinese herbal medicine with complex system,and the impacts of processing and pathological state on pharmacokinetics of Chinese herbal extracts,aiming to provide a reference for quality control,product development and rational medication of Chinese herbal extracts.


Subject(s)
Drugs, Chinese Herbal , China , Humans , Medicine, Chinese Traditional , Quality Control
11.
World J Clin Cases ; 9(22): 6485-6492, 2021 Aug 06.
Article in English | MEDLINE | ID: mdl-34435016

ABSTRACT

BACKGROUND: Hydrocephalus following dural tear after spinal surgery is rare. Although a few cases of obstructive hydrocephalus caused by subdural fluid collection and communicating hydrocephalus associated with meningitis have been reported, the mechanism remains uncertain. Herein we describe a patient complicated with hydrocephalus after cervical laminoplasty in whom subdural fluid collection in the cervical spine and posterior cranial fossa rather than chronic meningitis was the main mechanism. CASE SUMMARY: A 45-year-old man underwent cervical laminoplasty for cervical spondylotic myelopathy at a local hospital. Ten days postoperatively, a high fever occurred and magnetic resonance imaging (MRI) showed cerebrospinal fluid (CSF) leakage. Pseudomeningocele liquid test showed high levels of protein and white blood cell (WBC) count with negative bacterial culture. The patient was treated with short-term intravenous antibiotic and discharged with normal body temperature. The patient was uneventful during the first 8 mo follow-up although repeated MRI showed persistent pseudomeningocele. At the 9th mo postoperatively, the patient gradually presented with dizziness and headache accompanied by recurrent weakness of his left arm. Imaging examinations demonstrated hydrocephalus and a cystic lesion around the cervical spinal cord. CSF test from lumbar puncture indicated chronic meningitis. MRI on 1 d after pseudomeningocele drainage showed a significant decrease in the cystic volume, suggesting that the cystic lesion would be subdural fluid collection rather than adhesive arachnoiditis. After dural defect repair, the patient's symptoms completely resolved and hydrocephalus gradually disappeared. CSF analysis at the 21-mo follow-up revealed significantly decreased protein level and WBC count. CONCLUSION: Subdural fluid collection rather than meningitis contributes to the hydrocephalus formation after cervical laminoplasty.

12.
J Clin Pathol ; 74(11): 697-703, 2021 Nov.
Article in English | MEDLINE | ID: mdl-34011618

ABSTRACT

To describe the clincopathological features and evaluate risk factors of post-transplant lymphoproliferative disorder (PTLD) after allogeneic haematopoietic stem cell transplants (allo-HSCT), with comparison between paediatric and adult .Clinicopathological features of 81 cases of PTLD after allo-HSCT were analysed by histopatholgy, immunohistochemistry and in situ hybridisatioin.The cases included 58 males and 23 females with a median age of 26.7 years (range 6-55 years) and the PTLDs developed 1-60 months post-transplant (mean 5.9 months). The histological types indicated 10 cases of non-destructive PTLD, including 4 of plasmacytic hyperplasia, 5 of infectious mononucleosis and 1 of florid follicular hyperplasia. Fifty-six cases were polymorphic PTLD, and 15 were monomorphic PTLD, including thirteen of diffuse large B cell lymphoma, 1 of extranodal nasal type natural killer (NK)/T cell lymphoma and 1 of plasmablastic lymphoma. Foci and sheets of necrosis were observed in 31 cases. The infected ratio of Epstein-Barr virus (EBV) was 91.4%. Some cases were treated by reduction of immunosuppression, antiviral therapy, donor lymphocyte infusion or anti-CD20 monoclonal rituximab. Thirty-three cases died. Compared with that of adult, overall survival of paediatric recipient may be better.The first half year after allo-HSCT is very important for the development of PTLD. Type of PTLD, EBV infection and graft-versus-host disease are risk factors. The prognosis of PTLD is poor, and PTLD after allo-HSCT exhibits some features different from that after solid organ transplantation and some differences existing between adult and paediatric recipients.


Subject(s)
Epstein-Barr Virus Infections/complications , Hematopoietic Stem Cell Transplantation/adverse effects , Lymphoproliferative Disorders , Transplantation, Homologous/adverse effects , Adolescent , Adult , Allogeneic Cells/virology , Antigens, CD20 , Child , Epstein-Barr Virus Infections/physiopathology , Female , Herpesvirus 4, Human/isolation & purification , Herpesvirus 4, Human/pathogenicity , Humans , Lymphoproliferative Disorders/complications , Lymphoproliferative Disorders/drug therapy , Lymphoproliferative Disorders/mortality , Lymphoproliferative Disorders/pathology , Male , Middle Aged , Risk Factors , Rituximab/therapeutic use
13.
Zhongguo Zhong Yao Za Zhi ; 46(7): 1752-1762, 2021 Apr.
Article in Chinese | MEDLINE | ID: mdl-33982479

ABSTRACT

Traditional Chinese medicine(TCM) injections boast a definite efficacy and have been widely used in clinic. However, the problems in medication safety have been attracted increasing attention. Pharmacokinetics is of significance to guiding TCM injection administration regimen design and improving safety and effectiveness in clinical use. In recent years, with the improvement of ideas, technology and methods of TCM studies, the pharmacokinetic studies of TCM injections have been broadly performed, with a notable progress. This paper reviewed the advance in pharmacokinetics studies of TCM injections in recent ten years, which mainly focused on pre-clinical concentration-time course, distribution, metabolism and excretion in vivo based on analysis techniques, pharmacokinetic interactions of constitutes, impact of pathological state, pharmacokinetic interactions between TCM injection and chemical drugs, and clinical pharmacokinetics studies of TCM injections, in the expectation of providing reference for studies on quality control, product development and rational clinical use of TCM injections.


Subject(s)
Drugs, Chinese Herbal , Medicine, Chinese Traditional , Injections , Quality Control
14.
BMC Biol ; 19(1): 83, 2021 04 23.
Article in English | MEDLINE | ID: mdl-33892710

ABSTRACT

BACKGROUND: Defining the origin of genetic novelty is central to our understanding of the evolution of novel traits. Diversification among fatty acid desaturase (FAD) genes has played a fundamental role in the introduction of structural variation in fatty acyl derivatives. Because of its central role in generating diversity in insect semiochemicals, the FAD gene family has become a model to study how gene family expansions can contribute to the evolution of lineage-specific innovations. Here we used the codling moth (Cydia pomonella) as a study system to decipher the proximate mechanism underlying the production of the ∆8∆10 signature structure of olethreutine moths. Biosynthesis of the codling moth sex pheromone, (E8,E10)-dodecadienol (codlemone), involves two consecutive desaturation steps, the first of which is unusual in that it generates an E9 unsaturation. The second step is also atypical: it generates a conjugated diene system from the E9 monoene C12 intermediate via 1,4-desaturation. RESULTS: Here we describe the characterization of the FAD gene acting in codlemone biosynthesis. We identify 27 FAD genes corresponding to the various functional classes identified in insects and Lepidoptera. These genes are distributed across the C. pomonella genome in tandem arrays or isolated genes, indicating that the FAD repertoire consists of both ancient and recent duplications and expansions. Using transcriptomics, we show large divergence in expression domains: some genes appear ubiquitously expressed across tissue and developmental stages; others appear more restricted in their expression pattern. Functional assays using heterologous expression systems reveal that one gene, Cpo_CPRQ, which is prominently and exclusively expressed in the female pheromone gland, encodes an FAD that possesses both E9 and ∆8∆10 desaturation activities. Phylogenetically, Cpo_CPRQ clusters within the Lepidoptera-specific ∆10/∆11 clade of FADs, a classic reservoir of unusual desaturase activities in moths. CONCLUSIONS: Our integrative approach shows that the evolution of the signature pheromone structure of olethreutine moths relied on a gene belonging to an ancient gene expansion. Members of other expanded FAD subfamilies do not appear to play a role in chemical communication. This advises for caution when postulating the consequences of lineage-specific expansions based on genomics alone.


Subject(s)
Moths , Animals , Female , Flavin-Adenine Dinucleotide , Gene Duplication , Moths/genetics , Pheromones/genetics , Phylogeny
15.
J Chem Ecol ; 47(3): 248-264, 2021 Mar.
Article in English | MEDLINE | ID: mdl-33779878

ABSTRACT

The European grapevine moth, Lobesia botrana, uses (E,Z)-7,9-dodecadienyl acetate as its major sex pheromone component. Through in vivo labeling experiments we demonstrated that the doubly unsaturated pheromone component is produced by ∆11 desaturation of tetradecanoic acid, followed by chain shortening of (Z)-11-tetradecenoic acid to (Z)-9-dodecenoic acid, and subsequently introduction of the second double bond by an unknown ∆7 desaturase, before final reduction and acetylation. By sequencing and analyzing the transcriptome of female pheromone glands of L. botrana, we obtained 41 candidate genes that may be involved in sex pheromone production, including the genes encoding 17 fatty acyl desaturases, 13 fatty acyl reductases, 1 fatty acid synthase, 3 acyl-CoA oxidases, 1 acetyl-CoA carboxylase, 4 fatty acid transport proteins and 2 acyl-CoA binding proteins. A functional assay of desaturase and acyl-CoA oxidase gene candidates in yeast and insect cell (Sf9) heterologous expression systems revealed that Lbo_PPTQ encodes a ∆11 desaturase producing (Z)-11-tetradecenoic acid from tetradecanoic acid. Further, Lbo_31670 and Lbo_49602 encode two acyl-CoA oxidases that may produce (Z)-9-dodecenoic acid by chain shortening (Z)-11-tetradecenoic acid. The gene encoding the enzyme introducing the E7 double bond into (Z)-9-dodecenoic acid remains elusive even though we assayed 17 candidate desaturases in the two heterologous systems.


Subject(s)
Dodecanol/analogs & derivatives , Sex Attractants/biosynthesis , Acetyl-CoA Carboxylase/genetics , Acetyl-CoA Carboxylase/metabolism , Amino Acid Sequence , Animals , Fatty Acid Desaturases/genetics , Fatty Acid Desaturases/metabolism , Fatty Acid Synthases/genetics , Fatty Acid Synthases/metabolism , Fatty Acid Transport Proteins/genetics , Fatty Acid Transport Proteins/metabolism , Female , Gas Chromatography-Mass Spectrometry , Moths , Myristic Acid/metabolism , Oxidoreductases/genetics , Oxidoreductases/metabolism , Saccharomyces cerevisiae/metabolism , Sf9 Cells/metabolism , Transcriptome
16.
Biotechnol J ; 16(6): e2100004, 2021 Jun.
Article in English | MEDLINE | ID: mdl-33656777

ABSTRACT

The European corn borer (ECB) Ostrinia nubilalis is a widespread pest of cereals, particularly maize. Mating disruption with the sex pheromone is a potentially attractive method for managing this pest; however, chemical synthesis of pheromones requires expensive starting materials and catalysts and generates hazardous waste. The goal of this study was to develop a biotechnological method for the production of ECB sex pheromone. Our approach was to engineer the oleaginous yeast Yarrowia lipolytica to produce (Z)-11-tetradecenol (Z11-14:OH), which can then be chemically acetylated to (Z)-11-tetradecenyl acetate (Z11-14:OAc), the main pheromone component of the Z-race of O. nubilalis. First, a C14 platform strain with increased biosynthesis of myristoyl-CoA was obtained by introducing a point mutation into the α-subunit of fatty acid synthase, replacing isoleucine 1220 with phenylalanine (Fas2pI1220F ). The intracellular accumulation of myristic acid increased 8.4-fold. Next, fatty acyl-CoA desaturases (FAD) and fatty acyl-CoA reductases (FAR) from nine different species of Lepidoptera were screened in the C14 platform strain, individually and in combinations. A titer of 29.2 ± 1.6 mg L-1  Z11-14:OH was reached in small-scale cultivation with an optimal combination of a FAD (Lbo_PPTQ) from Lobesia botrana and FAR (HarFAR) from Helicoverpa armigera. When the second copies of FAD and FAR genes were introduced, the titer improved 2.1-fold. The native FAS1 gene's overexpression led to a further 1.5-fold titer increase, reaching 93.9 ± 11.7 mg L-1  in small-scale cultivation. When the same engineered strain was cultivated in controlled 1 L bioreactors in fed-batch mode, 188.1 ± 13.4 mg L-1  of Z11-14:OH was obtained. Fatty alcohols were extracted from the biomass and chemically acetylated to obtain Z11-14:OAc. Electroantennogram experiments showed that males of the Z-race of O. nubilalis were responsive to biologically-derived pheromone blend. Behavioral bioassays in a wind tunnel revealed attraction of male O. nubilalis, although full precopulatory behavior was observed less often than for the chemically synthesized pheromone blend. The study paves the way for the production of ECB pheromone by fermentation.


Subject(s)
Moths , Sex Attractants , Yarrowia , Yeast, Dried , Animals , Male , Moths/genetics , Yarrowia/genetics , Zea mays/genetics
17.
Environ Res ; 195: 110752, 2021 04.
Article in English | MEDLINE | ID: mdl-33485908

ABSTRACT

3-Nitro-4-hydroxy-phenylarsonic acid (3-NHPAA), an organic-arsenic compound, as one of widely used antibacterial veterinary drug, has greatly attracted the attention due to its potential threats on ecological environment. A series of the nanocomposites of zirconia nanoparticles with crystal phases (pure monoclinic, pure tetragonal and mixed phase (monoclinic + tetragonal)) anchored on reduced graphene oxide were produced through managing the concentration of triethanolamine solution and the reaction time. The effects of the crystal phases of the zirconia in the structure of the nanocomposites were played a key role in the adsorption performances of the 3-NHPAA. Experiment data identified the nanocomposites with monoclinic phase of zirconia excelled at the adsorption of the 3-NHPAA with a higher adsorption capacity up to 207.2 mg g-1. The uptake of the 3-NHPAA by the three nanocomposites was implemented within 60 min and highly pH-dependent which illustrated electrostatic attraction between them as a main mechanism during the adsorption process. A wider pH range (3.8-8.8) for the uptake of the 3-NHPAA by the nanocomposites with the monoclinic phase of zirconia was obtained compared with the nanocomposites containing tetragonal phase (3.8-5.9) or the mixed phase (3.8-7.1) of zirconia. The adsorption of the 3-NHPAA was well described by the pseudo-second order kinetic and Langmuir equations. The thermodynamic parameters suggested that the adsorption of the 3-NHPAA over the three nanocomposites was endothermic and spontaneous in nature. In summary, the nanocomposites of reduced graphene oxide and monoclinic phase of zirconia nanoparticles as an adsorbent were better to the adsorption of the 3-NHPAA.


Subject(s)
Arsenic , Graphite , Nanocomposites , Pharmaceutical Preparations , Water Pollutants, Chemical , Adsorption , Arsenic/analysis , Hydrogen-Ion Concentration , Kinetics , Water Pollutants, Chemical/analysis , Zirconium
18.
Hepatobiliary Pancreat Dis Int ; 20(4): 337-344, 2021 Aug.
Article in English | MEDLINE | ID: mdl-32933876

ABSTRACT

BACKGROUND: Anillin (ANLN) is required for tumor growth. It has been proven that knockdown of ANLN effectively reduces the occurrence of hepatocellular carcinoma (HCC) in transgenic mice. However, the functional role of ANLN in HCC patients remains to be elucidated. METHODS: Both microarray and TCGA project were used for the analyses of ANLN expression and regulation in HCC. The effect of ANLN on proliferation and cell cycle was detected by CCK-8, colony formation assay and flow cytometry. ANLN expression was measured by immunohistochemistry. Correlation between ANLN expression and clinicopathological features was assessed by Pearson Chi-square test and 5-year overall survival after liver resection was evaluated by Kaplan-Meier method. RESULTS: Increased copy number, decreased methylation levels in the CpG island and upregulated histone hypermethylation of ANLN were found in HCC. Knockdown of ANLN inhibited proliferation and induced G2/M phase arrest in SMMC-7721 cells. ANLN was mainly expressed in the nucleus and showed significantly higher expression levels in cancerous tissues than those in paired adjacent tissues. Moreover, nuclear ANLN expression levels in HCC metastases were significantly higher than those in primary HCC. The results of Cox proportional hazards regression model suggested that ANLN nuclear expression in HCC was an independent risk factor for poor 5-year overall survival of patients after liver resection. CONCLUSIONS: ANLN is a potential therapeutic target for HCC. Patients with nuclear ANLN overexpression in HCC tissue may need adjuvant therapy after liver resection.


Subject(s)
Carcinoma, Hepatocellular , Liver Neoplasms , Animals , Biomarkers, Tumor/genetics , Carcinoma, Hepatocellular/genetics , Carcinoma, Hepatocellular/surgery , Cell Line, Tumor , Cell Proliferation , Contractile Proteins , Gene Expression Regulation, Neoplastic , Humans , Kaplan-Meier Estimate , Liver Neoplasms/genetics , Liver Neoplasms/surgery , Mice , Prognosis
19.
Metab Eng ; 62: 312-321, 2020 11.
Article in English | MEDLINE | ID: mdl-33045365

ABSTRACT

The use of insect sex pheromones is an alternative technology for pest control in agriculture and forestry, which, in contrast to insecticides, does not have adverse effects on human health or environment and is efficient also against insecticide-resistant insect populations. Due to the high cost of chemically synthesized pheromones, mating disruption applications are currently primarily targeting higher value crops, such as fruits. Here we demonstrate a biotechnological method for the production of (Z)-hexadec-11-en-1-ol and (Z)-tetradec-9-en-1-ol, using engineered yeast cell factories. These unsaturated fatty alcohols are pheromone components or the immediate precursors of pheromone components of several economically important moth pests. Biosynthetic pathways towards several pheromones or their precursors were reconstructed in the oleaginous yeast Yarrowia lipolytica, which was further metabolically engineered for improved pheromone biosynthesis by decreasing fatty alcohol degradation and downregulating storage lipid accumulation. The sex pheromone of the cotton bollworm Helicoverpa armigera was produced by oxidation of fermented fatty alcohols into corresponding aldehydes. The resulting yeast-derived pheromone was just as efficient and specific for trapping of H. armigera male moths in cotton fields in Greece as a conventionally produced synthetic pheromone mixture. We further demonstrated the production of (Z)-tetradec-9-en-1-yl acetate, the main pheromone component of the fall armyworm Spodoptera frugiperda. Taken together our work describes a biotech platform for the production of commercially relevant titres of moth pheromones for pest control via yeast fermentation.


Subject(s)
Moths , Sex Attractants , Yarrowia , Animals , Fermentation , Humans , Male , Pest Control
20.
Ann Hematol ; 99(11): 2659-2670, 2020 Nov.
Article in English | MEDLINE | ID: mdl-32734550

ABSTRACT

Cytomegalovirus (CMV) can cause end-organ diseases including pneumonia, gastroenteritis, retinitis, and encephalitis in hematopoietic stem cell transplantation recipients. Potential differences among different CMV diseases remain uncertain. This study aimed to compare the clinical characteristics, risk factors, and mortality among different CMV diseases. A retrospective nested case-control study was performed based on a cohort of 3862 patients who underwent haploidentical hematopoietic stem cell transplantation at a single-center. CMV diseases occurred in 113 (2.92%) of 3862 haplo-HSCT recipients, including probable CMV pneumonia (CMVP, n = 34), proven CMV gastroenteritis (CMVG, n = 34), CMV retinitis (CMVR, n = 31), probable CMV encephalitis (CMVE, n = 7), and disseminated CMV disease (Di-CMVD, n = 7). Most (91.2%) cases of CMVG developed within 100 days, while most (90.3%) cases of CMVR were late onset. Refractory CMV infection and CMV viral load at different levels were associated with an increased risk of CMVP, CMVG, and CMVR. Compared with patients without CMV diseases, significantly higher non-relapse mortality at 1 year after transplantation was observed in patients with CMVP and CMVR, rather than CMVG. Patients with CMVP, Di-CMVD, and CMVE had higher overall mortality after diagnosis than that of patients with CMVG and CMVR (61.7%, 57.1%, 40.0% vs 27.7%, 18.6%, P = 0.001). In conclusion, the onset time, viral dynamics, and mortality differ among different CMV diseases. The mortality of CMV diseases remains high, especially for CMVP, Di-CMVD, and CMVE.


Subject(s)
Cytomegalovirus Infections , Cytomegalovirus , Hematopoietic Stem Cell Transplantation , Adolescent , Adult , Allografts , Child , Child, Preschool , Cytomegalovirus Infections/diagnosis , Cytomegalovirus Infections/etiology , Cytomegalovirus Infections/mortality , Cytomegalovirus Infections/therapy , Disease-Free Survival , Female , Humans , Male , Middle Aged , Retrospective Studies , Risk Factors , Survival Rate , Time Factors
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