Small Molecular Metabolites Research Articles

Halofuginone reduces the inflammatory responses of DSS-induced colitis through metabolic reprogramming.

Hypoxia and inflammation have been identified as the hallmarks of colitis, intertwined with metabolism. Here, we report that halofuginone (HF), an antiparasitic drug, attenuates dextran sulfate sodium (DSS)-induced colitis in mice, as represented by attenuating the disease activity index, inhibiting colonic shortening, ameliorating colonic lesions and histological signs of damage, reducing colonic myeloperoxidase activity, and suppressing the production of pro-inflammatory cytokines in colon tissue. Intriguingly, the hypoxia-inducible factor 1alpha (HIF-1α) and tumor necrosis factor alpha were also suppressed by HF treatment in colon tissues, exhibiting a tissue-specific effect. To further reveal the metabolic signatures upon HF treatment, mass spectrometry-based metabolomic analysis of the small molecular metabolites in liver, spleen and colon tissues was performed. As a result, we found that HF treatment counteracted the levels of acylcarnitines, including palmitoyl-l-carnitine, isobutyrylcarnitine, vaccenylcarnitine, and myristoylcarnitine, in colon tissues with DSS induction, but no significant change in the levels of acylcarnitines was observed in liver or spleen tissues. The metabolic signatures may indicate that incomplete fatty acid oxidation (FAO) in the colon could be restored upon HF treatment as the tissue-specific metabolic characterization. Taken together, our findings uncovered that the HF potentiated anti-inflammatory effect in DSS-induced colitis in mice and its underlying mechanisms could be associated with the inhibition of HIF-1α and reduced levels of acylcarnitines, suggesting that both the inhibition of HIF-1α and the counteraction of incomplete FAO might be useful in the prevention and treatment of inflammatory bowel disease.

An acetyl-L-carnitine switch on mitochondrial dysfunction and rescue in the metabolomics study on aluminum oxide nanoparticles.

BackgroundDue to the wide application of engineered aluminum oxide nanoparticles and increased aluminum containing particulate matter suspending in air, exposure of human to nano-scale aluminum oxide nanoparticles (Al2O3 NPs) is becoming inevitable.MethodsIn the present study, RNA microarray coupled with metabolomics analysis were used to uncover mechanisms underlying cellular responses to Al2O3 NPs and imply the potential rescue.ResultsWe found that Al2O3 NPs significantly triggered down-regulation of mitochondria-related genes located in complex I, IV and V, which were involved in oxidative phosphorylation and neural degeneration pathways, in human bronchial epithelial (HBE) cells. Subsequent cell- and animal- based assays confirmed that Al2O3 NPs caused mitochondria-dependent apoptosis and oxidative stress either in vitro or in vivo, which were consistent with the trends of gene regulation. To rescue the Al2O3 NPs induced mitochondria dysfunction, disruption of small molecular metabolites of HBE were profiled using metabolomics analysis, which facilitates identification of potential antagonizer or supplement against nanoparticle-involved damages. Supplementation of an antioxidant, acetyl-L-carnitine, completely or partially restored the Al2O3 NPs modulated gene expression levels in mitochondrial complex I, IV and V. It further reduced apoptosis and oxidative damages in both Al2O3 NPs treated HBE cells and animal lung tissues.ConclusionThus, our results demonstrate the potential mechanism of respiratory system damages induced by Al2O3 NPs. Meanwhile, based on the metabolomics profiling, application of acetyl-L-carnitine is suggested to ameliorate mitochondria dysfunction associated with Al2O3 NPs.Electronic supplementary materialThe online version of this article (doi:10.1186/s12989-016-0115-y) contains supplementary material, which is available to authorized users.

Effect of dietary sodium restriction on human urinary metabolomic profiles.

Metabolomics is a relatively new field of "-omics" research, focusing on high-throughput identification of small molecular weight metabolites. Diet has both acute and chronic effects on metabolic profiles; however, alterations in response to dietary sodium restriction (DSR) are completely unknown. The goal of this study was to explore changes in urine metabolites in response to DSR, as well as their association with previously reported improvements in vascular function with DSR. Using stored urine samples from a 10-week randomized placebo-controlled crossover study of DSR in 17 middle-aged/older adults (six men and 11 women; mean age 62±8 years) who had moderately elevated systolic BP (130-159 mmHg) and were otherwise healthy, a liquid chromatography/mass spectrometry-based analysis of 289 metabolites was performed. This study identified metabolites that were significantly altered between the typical (153±29 mmol/d) and low (70±29 mmol/d) sodium conditions, as well as their baseline (typical sodium) association with responsiveness to previously reported improvements in vascular endothelial function (brachial artery flow-mediated dilation) and large elastic artery stiffness (aortic pulse wave velocity). Of the 289 metabolites surveyed, 10 were significantly altered (nine were upregulated and one was downregulated) during the low sodium condition, and eight of these exceeded our prespecified clinically significant threshold of a >40% change. These metabolites were involved in biologic pathways broadly related to cardiovascular risk, nitric oxide production, oxidative stress, osmotic regulation, and metabolism. One metabolite, serine, was independently (positively) associated with previously reported improvements in the primary vascular outcome of brachial artery flow-mediated dilation. This proof-of-concept study provides the first evidence that DSR is a stimulus that induces significant changes in urinary metabolomic profiles. Moreover, serine was independently associated with corresponding changes in vascular endothelial function after DSR. Larger follow-up studies will be required to confirm and further elucidate the metabolic pathways that are altered in response to DSR.

Integrated Datasets Characterize Metabolic Interactions between Mouse's Colonic Mucosa, Colonic‐Cecal Contents and Feces

The pattern of metabolites produced by the gut microbiome comprises a phenotype indicative of the means by which that microbiome affects the gut. We characterized that phenotype by conducting metabolomic analyses of the colonic-cecal contents, comparing that to the metabolite patterns of feces and colonic mucosa from mice fed a standard chow diet. We detected >270 small molecular weight metabolites by gas chromatograph, time-of-flight mass spectrometry (GC-TOF) and ultra-high performance liquid chromatography, quadrapole time- of-flight mass spectrometry (UPLC-Q-TOF). Of that number 58 (21%) varied significantly between all three sample types, while 82 (30%) showed similar patterns in mucosa and feces, and 160 (58%) showed similar patterns in the colonic-cecal contents and either mucosa or feces. In these sample type comparisons, metabolic pathway enrichment analysis revealed that these detected 275-metabolites can be categorized into 28 biochemical pathways as defined by KEGG, and among which 14 pathways are common to all sample types. These results comprise the first characterization of relationships among metabolites in the colonic-cecal contents, feces and colonic mucosa in a healthy mouse model. This information provides the groundwork for investigating metabolic interactions in the gut.

Comparison between moving bed-membrane bioreactor and conventional membrane bioreactor systems. Part I: membrane fouling

The combination of moving bed biofilm reactor and traditional membrane bioreactor (MBR) shows great potential to be widely used in Liaohe River basin for wastewater treatment. The aim of this research is to compare the bioreactor performance and membrane fouling between integrated fixed-film activated sludge membrane bioreactor (IFAS–MBR) and MBR. Two lab-scale reactors were operated in parallel to treat synthetic municipal wastewater with continuous operation. The results indicated that both systems had the same high removal efficiency of ammonium, while the IFAS–MBR showed a higher ability to remove COD. The trans-membrane pressure in the MBR was higher than that in the IFAS–MBR during the entire operation, due to a higher total modified fouling index of the mixed liquor and the resultant higher membrane fouling potential. The concentrations of extracellular polymeric substances and soluble microbial products were higher in MBR, despite the functional groups were almost similar in two reactors. The conventional MBR had more small molecular weight components (<1 kDa) and soluble microbial metabolites. This seems to cause pore blocking, which directly results in membrane flux declining and membrane resistance rising in the MBR system. In addition, the MBR was characterized by a serious membrane fouling as more dense deposits were observed by scanning electron microscopy. On the contrary, the IFAS–MBR showed a lower membrane fouling tendency during the whole operation.

Metabolomic analysis of urine samples of vivax malaria in-patients for biomarker identification

Plasmodium vivax is the most dominating species in the Indian subcontinent among the five Plasmodium species causing malarial infection in humans. This species is generally believed to be benign and classically thought to cause uncomplicated clinical infection that rarely leads to death or other complication. However, recent studies have shown incidences of P. vivax infection leading to severe complications and even death. To understand the biochemical changes at the pathway level and the modality through which the host recovers from the infection, we monitored the small molecular weight metabolites of urine samples of hospital in-patients undergoing treatment for vivax malaria. Metabolomics, involving 1H NMR spectra of urine samples subjected to multivariate statistical analyses, was employed for this purpose. Urine metabolic profiles were used to identify putative biomarkers and establishing correlation with parasite count in the peripheral blood. The untreated malaria patients (day1) exhibited altered excretion of taurine, hippurate, citrate, glycine, 3-methylhistidine and alanine in comparison to the self-control profile on day30. The day1–4 urine metabolite profiles are distinct from day5 and day30 profiles. The latter two profiles are indistinguishable. Temporal trajectories of urinary metabolites indicated an inflection point on day3. The day1 1H NMR profiles, distinct from the profiles of viral fever patients, correlated with the parasitemia levels in the patients and could in fact be used to gauge the anemia status of the patients. This study demonstrates the potential of urine metabolic profiling in understanding the pathophysiology of P. vivax infection and in non-invasive disease diagnosis.

Plasma kinetics and microbial biotransformation of grape seed flavanols in rats

Flavan-3-ols and their oligomeric forms, proanthocyanidins (PAs), are poorly absorbed in the small intestine and reach the colon where gut bacteria enzymes can hydrolyse them to produce small molecular metabolites, which can reach systemic circulation. However, the microbial metabolism of flavanols has been poorly described in vivo. The aim of this study was to determine the colonic biotransformation pathway and the plasma temporal appearance of grape seed flavanols colonic metabolites in rats. Rat plasma colonic metabolites were analysed by HPLC-MS/MS at 2, 7, 24 and 48 h after 1000 mg/kg of a grape seed PA extract (GSPE) administration. Results indicated that non-metabolised flavanols have peak plasma concentrations 2 h after GSPE administration, whereas the colonic metabolites appeared in plasma later, indicating their gradual colonic biotransformation as valerolactone > phenylpropionic acids ≈ phenylacetic acids > benzoic acids. This study shows how flavanols are biotransformed by gut bacteria in rats over time, facilitating potential bioactive compound identification for particular health effects.

Search for biomarkers in critically ill patients: a new approach based on nuclear magnetic resonance spectroscopy of mini-bronchoalveolar lavage fluid

AbstractNo abstract

Flavonoids as important molecules of plant interactions with the environment.

Flavonoids are small molecular secondary metabolites synthesized by plants with various biological activities. Due to their physical and biochemical properties, they are capable of participating in plants’ interactions with other organisms (microorganisms, animals and other plants) and their reactions to environmental stresses. The majority of their functions result from their strong antioxidative properties. Although an increasing number of studies focus on the application of flavonoids in medicine or the food industry, their relevance for the plants themselves also deserves extensive investigations. This review summarizes the current knowledge on the functions of flavonoids in the physiology of plants and their relations with the environment.

A derivatization and validation strategy for determining the spatial localization of endogenous amine metabolites in tissues using MALDI imaging mass spectrometry.

Imaging mass spectrometry (IMS) studies increasingly focus on endogenous small molecular weight metabolites and consequently bring special analytical challenges. Since analytical tissue blanks do not exist for endogenous metabolites, careful consideration must be given to confirm molecular identity. Here, we present approaches for the improvement in detection of endogenous amine metabolites such as amino acids and neurotransmitters in tissues through chemical derivatization and matrix-assisted laser desorption/ionization (MALDI) IMS. Chemical derivatization with 4-hydroxy-3-methoxycinnamaldehyde (CA) was used to improve sensitivity and specificity. CA was applied to the tissue via MALDI sample targets precoated with a mixture of derivatization reagent and ferulic acid as a MALDI matrix. Spatial distributions of chemically derivatized endogenous metabolites in tissue were determined by high-mass resolution and MS(n) IMS. We highlight an analytical strategy for metabolite validation whereby tissue extracts are analyzed by high-performance liquid chromatography (HPLC)-MS/MS to unambiguously identify metabolites and distinguish them from isobaric compounds.

Metabolic profiling of human lung injury by 1H high-resolution nuclear magnetic resonance spectroscopy of blood serum

Acute respiratory distress syndrome (ARDS) contributes substantially to mortality and morbidity in USA and worldwide. Due to limitations in early diagnostics of ARDS by classical methods, there has been need for discovery of novel methods and biomarkers for its characterization. We present here first high-resolution 1H nuclear magnetic resonance (NMR) metabolic profiling of serum from ARDS patients and non ARDS (NARDS) controls to search for novel biomarkers in blood serum for better diagnostics and prognostics. We have carried out study with serum samples from a total of 45 subjects, which included 26 ARDS patients and 19 NARDS controls. Principal component analysis (PCA) and partial least square discriminant analysis (PLS-DA) were performed on 1H NMR spectra of serum for group discrimination between the two. PCA and PLS-DA on 1H NMR spectra of serum clearly discriminated between NARDS control and ARDS diseased samples. In our study, lipid resonances showed major contribution for this separation in loading plots. In order to highlight role of other small molecular weight metabolites, the analysis was also carried out after removing lipids resonances from NMR spectrum. There was significant increase in concentrations of N-acetylglycoproteins (NAC) (p = 0.001), acetoacetate (p = 0.001), lactate (p = 0.001), creatinine (p = 0.003), histidine (p = 0.03), formate (p = 0.04) and aromatic amino acids serum of ARDS patients. Along with small metabolites, lipids play a very important role in this discrimination and can differentiate between two classes. Our study has given new avenue in the metabolic profiling of lung injuries.

Interface solution isoelectric focusing with in situ MALDI-TOF mass spectrometry.

This paper describes a simple and reusable microfluidic device combining solution IEF (sIEF) with MALDI-TOF MS for rapid proteomic and metabolic analysis of microliter samples. The device contains two glass plates with nanoliter microwell arrays, which can be assembled to form a fluidic path for sIEF separation, and reconfigured for dividing separated bands. One microliter samples can be loaded and separated by sIEF into static bands in 10∼30 min. After a slipping operation, the static IEF bands can be divided into nanoliter droplets in microwells without mobilization, and the device can be opened for in situ MALDI-TOF MS detection without loss of separation resolution. The performance of the device is characterized by separating and identifying intact proteins. The applicability in metabolic analysis is demonstrated by preliminary experiments on profiling small molecular metabolites in cerebrospinal fluid microdialysates from rat brain.

The role of 3 external quality assessment programs in assessment of analytical quality on determination of serum creatinine and urea

Objective To describe and compare the roles of 3 external quality assessment programs in assessment of analytical quality of serum creatinine and urea. Methods Research in quality management methods.Sixty-five laboratories those enrolled in the Natonal Center for Clinical Laboratories′ programs of routine chemistry external quality assessment (EQA), trueness verification (TV) for small molecular metabolites and external comparison of internal quality control (IQC) simultaneously in 2013 were selected，the performances of those laboratories of serum creatinine (crea) and urea in terms of total errors(TE), bias and CV were obtained by using the above 3 programs, and these performance were assessed against the criterion listed in the analytical quality specifications for routine analysis in clinical biochemistry (WS/T 403-2012). The failure ratio of 65 laboratories on each performance was calculated, the sensitivity of 3 external quality assessment programs in detection of analytical quality deficiency among clinical laboratories were compared. Results Only 1 laboratory failed in the 1st routine chemistry EQA in terms of TE of creatinine, failure ratio is 1.5%(1/65). Three laboratories failed in the 2nd EQA and caused a failure ratio of 4.6%(3/65). For serum urea, 3 laboratories failed in the 1st routine chemistry EQA with a failure ratio of 4.6%(3/65). Two laboratories failed in the 2nd EQA with a failure ratio of 3.1%(2/65).The failure ratios of creatinine determination in two samples in TV were 41.5%(24/65) and 21.5%(14/65) respectively, and the failure ratio of urea determination were 53.8%(36/65) and 32.3%(21/65) respectively. In the program of external comparison of IQC，the CVs of creatinine and urea determination ranged from 0.7% to 6.2% and from 1.0% to 7.2% respectively, their respective failure ratio range were 15.4%(10/65) and 40.0%(26/65). The failure ratio in routine EQA were much less than those in the other two programs, the laboratories failed in routine EQA program were all failed in trueness verification or/and the comparison of IQC programs, but not vice versa. Conclusions By participating in the programs of routine EQA, TV and comparison of IQC laboratories could assess the performances of inaccuracy, bias and imprecision. Laboratories should participate in different external quality assurance programs to detect their quality issues and get improved.（Chin J Lab Med,2014,37:357-361） Key words: Laboratories, hospital; Quality control; Creatinine; Urea; Uncertainty

Can metabolomics improve tuberculosis diagnostics?

With one third of the global human population suffering from tuberculosis (TB) in 2001, the disease was officially declared a worldwide emergency by the World Health Organization. Since then this disease has grown to epidemic proportions and the recent emergence of drug resistance, the rising incidence of HIV/TB co-infection, and the inability of the currently used vaccination, diagnostic and treatment protocols to control this pandemic, has made TB a research topic inviting urgent attention. The implementation of metabolomics, a relatively new research approach, which is defined as the study of all the small molecular weight compounds or metabolites in a system or sample, using highly sensitive and specific analytical techniques, shows promise in the quest to eradicate this disease. In this context, we describe here the advantages and limitations of the currently available TB diagnostic techniques, and the role that metabolomics has played in the identification of new biomarkers, not only leading to innovative approaches for TB diagnostics, but also to a better understanding of the intra-host changes induced by Mycobacterium tuberculosis infection and active disease.

Potential Applications and Development of Cell Metabolomics in Natural Products

Natural products play a vital role in drug discovery. Metabolomics, the systematic analysis of small molecular metabolites in biological systems, has been increasingly applied to discovering biomarkers, identifying perturbed pathways and discovering new drugs. It enables the examination and identification of endogenous biochemical reaction products occurring within a living cell. Cell metabolomics has many potential applications and advantages to currently used methods at various aspects in the natural product era. This review covers recent developments in the use of metabolomics to characterize cellular metabolome and interpret them in terms of metabolic changes taking place in a wide range of situations, especially, highlights the key roles of the endogenous small-molecule metabolites in the natural product field.

Microscopic imaging mass spectrometry assisted by on‐tissue chemical derivatization for visualizing multiple amino acids in human colon cancer xenografts

Imaging MS combined with CE/MS serves as a method to provide semi-quantitative and spatial information of small molecular metabolites in tissue slices. However, not all metabolites including amino acids have fully been visualized, because of low-ionization efficiency in MALDI MS. This study aimed to acquire semi-quantitative spatial information for multiple amino acids in frozen tissue slices. As a derivatization reagent, p-N,N,N-trimethylammonioanilyl N'-hydroxysuccinimidyl carbamate iodide (TAHS) was applied to increase their ionization efficiency and detection sensitivity. Semi-quantitative MALDI-imaging MS allowed us to visualize and quantify free amino acid pools in human colon cancer xenografts using a model of liver metastases in super-immunodeficient NOD/scid/γ(null) mice (NOG mice). Because the m/z values of several TAHS-derivatized amino acids overlap with those of the 2,5-dihydroxybenzoic acid background and other endogenous compounds, we imaged them with tandem MS. The results indicated that regional contents of glutamate, glutamine, glycine, leucine/isoleucine/hydroxyproline, phenylalanine, and alanine were significantly elevated in metastatic tumors versus parenchyma of tumor-bearing livers. On-tissue TAHS derivatization thus serves as a useful method to detect alterations in many amino acid levels in vivo, thereby enabling understanding of the spatial alterations of these metabolites under varied disease conditions including cancer.

Metabolism Studies of Unformulated Internally [3H]-Labeled Short Interfering RNAs in Mice

Absorption, distribution, metabolism, and excretion properties of two unformulated model short interfering RNA (siRNAs) were determined using a single internal [(3)H]-radiolabeling procedure, in which the full-length oligonucleotides were radiolabeled by Br/(3)H -exchange. Tissue distribution, excretion, and mass balance of radioactivity were investigated in male CD-1 mice after a single intravenous administration of the [(3)H]siRNAs, at a target dose level of 5 mg/kg. Quantitative whole-body autoradiography and liquid scintillation counting techniques were used to determine tissue distribution. Radiochromatogram profiles were determined in plasma, tissue extracts, and urine. Metabolites were separated by liquid chromatography and identified by radiodetection and high-resolution accurate mass spectrometry. In general, there was little difference in the distribution of total radiolabeled components after administration of the two unformulated [(3)H]siRNAs. The radioactivity was rapidly and widely distributed throughout the body and remained detectable in all tissues investigated at later time points (24 and 48 hours for [(3)H]MRP4 (multidrug resistance protein isoform 4) and [(3)H]SSB (Sjögren Syndrome antigen B) siRNA, respectively). After an initial rapid decrease, concentrations of total radiolabeled components in dried blood decreased at a much slower rate. A nearly complete mass balance was obtained for the [(3)H]SSB siRNA, and renal excretion was the main route of elimination (38%). The metabolism of the two model siRNAs was rapid and extensive. Five minutes after administration, no parent compound could be detected in plasma. Instead, radiolabeled nucleosides resulting from nuclease hydrolysis were observed. In the metabolism profiles obtained from various tissues, only radiolabeled nucleosides were found, suggesting that siRNAs are rapidly metabolized and that the distribution pattern of total radiolabeled components can be ascribed to small molecular weight metabolites.

Hippocampal metabolomics using ultrahigh-resolution mass spectrometry reveals neuroinflammation from Alzheimer’s disease in CRND8 mice

In the wake of genomics, metabolomics characterizes the small molecular metabolites revealing the phenotypes induced by gene mutants. To address the metabolic signatures in the hippocampus of the amyloid-beta (Aβ) peptides produced in transgenic (Tg) CRND8 mice, high-field ion cyclotron resonance-Fourier transform mass spectrometry supported by LC-LTQ-Orbitrap was introduced to profile the extracted metabolites. More than 10,000 ions were detected in the mass profile for each sample. Subsequently, peak alignment and the 80% rule followed by feature selection based on T score computation were performed. The putative identification was also conducted using the highly accurate masses with isotopic distribution by interfacing the MassTRIX database as well as MS/MS fragmentation generated in the LTQ-Orbitrap after chromatographic separation. Consequently, 58 differentiating masses were tentatively identified while up to 44 differentiating elemental compositions could not be biologically annotated in the databases. Nonetheless, of the putatively annotated masses, eicosanoids in arachidonic acid metabolism, fatty acid beta-oxidation disorders as well as disturbed glucose metabolism were highlighted as metabolic traits of Aβ toxicity in Tg CRND8 mice. Furthermore, a web-based bioinformatic tool was used for simulation of the metabolic pathways. As a result of the obtained metabolic signatures, the arachidonic acid metabolism dominates the metabolic perturbation in hippocampal tissues of Tg CRND8 mice compared to non-Tg littermates, indicating that Aβ toxicity functions neuroinflammation in hippocampal tissue and new theranostic opportunities might be offered by characterization of altered arachidonic acid metabolism for Alzheimer's disease.

Biomarkers of primary dysmenorrhea and herbal formula intervention: an exploratory metabonomics study of blood plasma and urine

Primary dysmenorrhea (PDM), a common clinical endocrine disorder affecting young women, is associated with endocrinopathy and metabolic abnormalities. Although some physiological and pathological function parameters have been investigated, little information about the changes of small metabolites in biofluids has been reported, which may cause poor diagnosis and treatment for PDM. The Xiang-Fu-Si-Wu Formula (XFSWF) is a Chinese herbal formula used to treat PDM for hundreds of years. The aim of this study was to establish the metabolic profile of PDM and investigate the action mechanism of XFSWF effect. In this cross-sectional study of 25 patients with PDM and 12 healthy controls, contents of small molecular endogenous metabolites in blood plasma and urine samples were measured by ultra performance liquid chromatography (UPLC) coupled with quadrupole-time-of-flight mass spectrometry (QTOF/MS) and triple quadrupole mass spectrometry (QqQ/MS) based techniques and analyzed by multivariate statistical methods. The levels of LPCs including lypso (16 : 1), lysoPC(20 : 4), lysoPC(18 : 2), lysoPC(16 : 0), lysoPC(18 : 1), lysoPC(10 : 1), estrone, 17-hydroxyprogesterone, myristoylglycine and palmitoylglycine increased significantly (p < 0.05) in PDM, while the levels of phytosphingosine, dihydrocortisol and sphingosine decreased significantly (p < 0.05) compared with the healthy controls. These significant perturbations are involved in glycerophospholipid metabolism and sphingolipid metabolism, as well as steroid hormone biosynthesis. The metabolic deviations recovered to the normal level after XFSWF intervention. The results demonstrated that biofluids metabonomics was a powerful tool in clinical diagnosis and treatment of PDM for providing information on changes in metabolites and neural, endocrinal and immune pathways. XFSWF can be used for the treatment of PDM cases, especially for those adolescents who do not desire a contraceptive method, to reduce the risk of secondary dysmenorrhea.

Biotechnology of non-Saccharomyces yeasts—the ascomycetes

Saccharomyces cerevisiae and several other yeast species are among the most important groups of biotechnological organisms. S. cerevisiae and closely related ascomycetous yeasts are the major producer of biotechnology products worldwide, exceeding other groups of industrial microorganisms in productivity and economic revenues. Traditional industrial attributes of the S. cerevisiae group include their primary roles in food fermentations such as beers, cider, wines, sake, distilled spirits, bakery products, cheese, sausages, and other fermented foods. Other long-standing industrial processes involving S. cerevisae yeasts are production of fuel ethanol, single-cell protein (SCP), feeds and fodder, industrial enzymes, and small molecular weight metabolites. More recently, non-Saccharomyces yeasts (non-conventional yeasts) have been utilized as industrial organisms for a variety of biotechnological roles. Non-Saccharomyces yeasts are increasingly being used as hosts for expression of proteins, biocatalysts and multi-enzyme pathways for the synthesis of fine chemicals and small molecular weight compounds of medicinal and nutritional importance. Non-Saccharomyces yeasts also have important roles in agriculture as agents of biocontrol, bioremediation, and as indicators of environmental quality. Several of these products and processes have reached commercial utility, while others are in advanced development. The objective of this mini-review is to describe processes currently used by industry and those in developmental stages and close to commercialization primarily from non-Saccharomyces yeasts with an emphasis on new opportunities. The utility of S. cerevisiae in heterologous production of selected products is also described.