Predicting Anaerobic Membrane Bioreactor Performance Using Flow-Cytometry-Derived High and Low Nucleic Acid Content Cells.

Having a tool to monitor the microbial abundances rapidly and to utilize the data to predict the reactor performance would facilitate the operation of an anaerobic membrane bioreactor (AnMBR). This study aims to achieve the aforementioned scenario by developing a linear regression model that incorporates a time-lagging mode. The model uses low nucleic acid (LNA) cell numbers and the ratio of high nucleic acid (HNA) to LNA cells as an input data set. First, the model was trained using data sets obtained from a 35 L pilot-scale AnMBR. The model was able to predict the chemical oxygen demand (COD) removal efficiency and methane production 3.5 days in advance. Subsequent validation of the model using flow cytometry (FCM)-derived data (at time t - 3.5 days) obtained from another biologically independent reactor did not exhibit any substantial difference between predicted and actual measurements of reactor performance at time t. Further cell sorting, 16S rRNA gene sequencing, and correlation analysis partly attributed this accurate prediction to HNA genera (e.g., Anaerovibrio and unclassified Bacteroidales) and LNA genera (e.g., Achromobacter, Ochrobactrum, and unclassified Anaerolineae). In summary, our findings suggest that HNA and LNA cell routine enumeration, along with the trained model, can derive a fast approach to predict the AnMBR performance.

Group theoretic particle swarm optimization for multi-level threshold lung cancer image segmentation.

Background: Image segmentation is an important step during the processing of medical images. For example, for the computer aid diagnostic systems for lung cancer image analysis, the segmented regions of tumors would help doctors in early diagnosis to determine timely and appropriate treatment possibilities and thereby improve the survival rate of the patients. However, general clinical routines of manual segmentation for large number of medical images are very difficult and time consuming, which is the challenge we aim to tackle using our proposed method. Methods: A novel image segmentation method with evolutionary learning technique named Group Theoretic Particle Swarm Optimization is proposed. It can tackle multi-level thresholding optimization problem during the segmentation process and rebuild the search paradigm according to the solid mathematical foundation of symmetric group from four designable aspects, which are particle encoding, solution landscape, neighborhood movement and swarm topology, respectively. The Kapur's entropy of multi-level thresholds is assessed as the objective function. Results: In contrast to those conventional metaheuristics methods for lung cancer image segmentation, this newly presented method generates the best performance result among them. Experimental results show that its Kapur's entropy has the value of 9.07, which is 16% higher than the worst case. Computational time is acceptable at the cost of 173.730 seconds, average level of evaluation metrics [Kappa, Precision, Recall, F1-measure, intersection over union (IoU) and receiver operating characteristic (ROC)] is over 90%, and search process of multi-level threshold combination would finally converge in the later phase of iterations after 700. The ablation study indicates that all components are significant to the contributions of our proposed method. Conclusions: Group Theoretic Particle Swarm Optimization for multi-level threshold segmentation is an efficient way to split a medical image into distinct regions and extract tumor tissues regions from the background. It maintains the balanced relationship between diversification and intensification during the search process and helps clinicians to make the diagnosis more accurately. Our proposed method processes potential medical value and clinical meanings.

Finite Element Analysis and Comparative Study of 4 Kinds of Internal Fixation Systems for Anterior Cervical Discectomy and Fusion in Children.

Background: Spinal injury in children usually occurs in the cervical spine region. Anterior fixation of the lower cervical spine has been applied in treating pediatric cervical spine injury and disease due to its stable and firm mechanical properties. This study performed finite element analysis and comparison of four different anterior cervical internal fixation systems for children to explore more standard methods of anterior cervical internal fixation in children and seek more effective and safe treatment for children's cervical spine diseases. Methods: A finite element model of 6-year-old children with lower cervical spine C4/5 discectomy was established, and the self-designed lower cervical spine anterior locking internal fixation system ACBLP and the children's anterior cervical internal fixation system ACOP, ACVLP, and ACSLP plate screws were fixed and loaded on the model. 27.42 N·m torque load was applied to each internal fixation model under six working conditions of anteflexion, backward flexion, left flexion, right flexion, left rotation, and right rotation, to simulate the movement of the cervical spine. The activity and stress distribution cloud diagram of each finite element model was obtained to explore the optimal method of anterior cervical fixation in children. Results: In the four internal fixation models of ACOP, ACVLP, ACSLP, and ACBLP, the mobility of the C4/5 segment showed a decreasing relationship, and the mobility of adjacent segments increased significantly. In the Mises stress cloud diagram of the cervical spine of the four models, the vertebral body and accessories of the ACBLP model born the least stress, followed by ACSLP. The steel plate and screws in the ACVLP internal fixation model were the most stressed. The stress of the internal fixation system (plate/screw) in all models increased in the order of ACBLP, ACSLP, ACVLP, and ACOP. Conclusions: ACBLP internal fixation system had obvious advantages in anterior internal fixation of the lower cervical spine in children, C4/5 had the smallest degree of movement, relative displacement was minimal, and the stress on the centrum and pedicle was the least, while the stress on the plate screw was relatively the smallest.

Attached-growth configuration outperforms continuously stirred tank anaerobic membrane bioreactors in alleviating membrane biofouling.

Biofouling impedes the performance of anaerobic membrane bioreactors (AnMBR). Two reactors, one as an up-flow attachment-growth AnMBR (UA-AnMBR) configuration, and the other, as a continuously stirred AnMBR (CS-AnMBR) were evaluated for differences in membrane fouling rate. TMP increment in UA-AnMBR was slower than CS-AnMBR, although both reactors had similar COD removal efficiency (ca. > 96%). Slower fouling rate for UA-AnMBR was related to lower total and viable cells, and thereby microbial activity compared to that in CS-AnMBR. Acinetobacter and Methanobacterium that played keystone roles in anaerobic biofilm formation were not consistently prevalent on the membranes connected to UA-AnMBR. This is in contrast to both Acinetobacter and Methanobacterium consistently prevalent on the membranes connected to CS-AnMBR. The findings suggest that UA-AnMBR can alleviate membrane biofouling through changes in microbial activity and profile dynamics, and would be a suitable reactor configuration to adopt to achieve an efficient AnMBR for municipal wastewater treatment.

Estimating the minimum number of SARS-CoV-2 infected cases needed to detect viral RNA in wastewater: To what extent of the outbreak can surveillance of wastewater tell us?

There is increasing interest in wastewater-based epidemiology (WBE) of SARS-CoV-2 RNA to serve as an early warning system for a community. Despite successful detection of SARS-CoV-2 RNA in wastewaters sampled from multiple locations, there is still no clear idea on the minimal number of cases in a community that are associated with a positive detection of the virus in wastewater. To address this knowledge gap, we sampled wastewaters from a septic tank (n = 57) and biological activated sludge tank (n = 52) located on-site of a hospital. The hospital is providing treatment for SARS-CoV-2 infected patients, with the number of hospitalized patients per day known. It was observed that depending on which nucleocapsid gene is targeted by means of RT-qPCR, a range of 253-409 positive cases out of 10,000 persons are required prior to detecting RNA SARS-CoV-2 in wastewater. There was a weak correlation between N1 and N2 gene abundances in wastewater with the number of hospitalized cases. This correlation was however not observed for N3 gene. The frequency of detecting N1 and N2 gene in wastewater was also higher than that for N3 gene. Furthermore, nucleocapsid genes of SARS-CoV-2 were detected at lower frequency in the partially treated wastewater than in the septic tank. In particular, N1 gene abundance was associated with water quality parameters such as total organic carbon and pH. In instances of positive detection, the average abundance of N1 and N3 genes in the activated sludge tank were reduced by 50 and 70% of the levels detected in septic tank, suggesting degradation of the SARS-CoV-2 gene fragments already occurring in the early stages of the wastewater treatment process.

Unraveling intrinsic correlation effects with angle-resolved photoemission spectroscopy.

Interaction effects can change materials properties in intriguing ways, and they have, in general, a huge impact on electronic spectra. In particular, satellites in photoemission spectra are pure many-body effects, and their study is of increasing interest in both experiment and theory. However, the intrinsic spectral function is only a part of a measured spectrum, and it is notoriously difficult to extract this information, even for simple metals. Our joint experimental and theoretical study of the prototypical simple metal aluminum demonstrates how intrinsic satellite spectra can be extracted from measured data using angular resolution in photoemission. A nondispersing satellite is detected and explained by electron-electron interactions and the thermal motion of the atoms. Additional nondispersing intensity comes from the inelastic scattering of the outgoing photoelectron. The ideal intrinsic spectral function, instead, has satellites that disperse both in energy and in shape. Theory and the information extracted from experiment describe these features with very good agreement.

Anharmonicity and Doping Melt the Charge Density Wave in Single-Layer TiSe2.

Low-dimensional systems with a vanishing band gap and a large electron-hole interaction have been proposed to be unstable toward exciton formation. As the exciton binding energy increases in low dimension, conventional wisdom suggests that excitonic insulators should be more stable in 2D than in 3D. Here we study the effects of the electron-hole interaction and anharmonicity in single-layer TiSe2. We find that, contrary to the bulk case and to the generally accepted picture, in single-layer TiSe2, the electron-hole exchange interaction is much smaller in 2D than in 3D and it has weak effects on phonon spectra. By calculating anharmonic phonon spectra within the stochastic self-consistent harmonic approximation, we obtain TCDW ≈ 440 K for an isolated and undoped single layer and TCDW ≈ 364 K for an electron-doping n = 4.6 × 1013 cm-2, close to the experimental result of 200-280 K on supported samples. Our work demonstrates that anharmonicity and doping melt the charge density wave in single-layer TiSe2.

Birefringent Fourier transform imaging spectrometer with a rotating retroreflector.

A birefringent Fourier transform imaging spectrometer with a new lateral shearing interferometer is presented. The interferometer includes a Wollaston prism and a retroreflector. It splits an incident light beam into two shearing parallel parts to obtain interference fringe patterns of an imaging target, which is well established as an aid in reducing problems associated with optical alignment and manufacturing precision. Continuously rotating the retroreflector enables the spectrometer to acquire two-dimensional spectral images without spatial scanning. This technology, with a high work efficiency and low complexity, is inherently compact and robust. The effectiveness of the proposed method is demonstrated by the experimental results.

Dynamical effects in electron spectroscopy.

One of the big challenges of theoretical condensed-matter physics is the description, understanding, and prediction of the effects of the Coulomb interaction on materials properties. In electronic spectra, the Coulomb interaction causes a renormalization of energies and change of spectral weight. Most importantly, it can lead to new structures, often called satellites. These can be linked to the coupling of excitations, also termed dynamical effects. State-of-the-art methods in the framework of many-body perturbation theory, in particular, the widely used GW approximation, often fail to describe satellite spectra. Instead, approaches based on a picture of electron-boson coupling such as the cumulant expansion are promising for the description of plasmon satellites. In this work, we give a unified derivation of the GW approximation and the cumulant expansion for the one-body Green's function. Using the example of bulk sodium, we compare the resulting spectral functions both in the valence and in the core region, and we discuss the dispersion of quasi-particles and satellites. We show that self-consistency is crucial to obtain meaningful results, in particular, at large binding energies. Very good agreement with experiment is obtained when the intrinsic spectral function is corrected for extrinsic and interference effects. Finally, we sketch how one can approach the problem in the case of the two-body Green's function, and we discuss the cancellation of various dynamical effects that occur in that case.

[Analysis of clinical effects of percutaneous vertebroplasty and percutaneous kyphoplasty in treating osteoporotic vertebral compression fracture].

OBJECTIVE: To explore the clinical outcomes of percutaneous vertebroplasty (PVP) and percutaneous kyphoplasty (PKP) in treating osteoporotic vertebral compression fracture (OVCF). METHODS: From January 2007 to February 2010, the data of 40 patients with osteoporotic vertebral compression fracture underwent treatment were retrospectively analyzed. Of them,20 patients were treated with PVP (PVP group), there were 8 males and 12 females with an average age of (66.37 +/- 2.34) years old (54 to 81); 20 patients were treated with PKP (PKP group), there were 11 males and 9 females with an average of (65.12 +/- 3.21) years old (56 to 79). Postoperative at 1 week, 12 weeks, 1 year, pain and daily life function were respectively assessed by visual analogue scale (VAS) and Barthel index (BI); and anterior height of responsibility vertebra, Cobb angle were measured by X-rays. RESULTS: In PVP group, 1 case complicated with bone cement leakage without clinical symptoms and no operation to treat. No postoperative infection and deep vein thrombosis were found between two groups. All patients were followed up more than 1 year, pain and daily life function has obviously improved than preoperative (P < 0.01); and there was no significant difference on 1 week, 12 weeks, 1 year after operation (P > 0.05); there was no significant difference between two groups (P > 0.05). In PVP group, there was no significant difference in anterior height of responsibility vertebra, Cobb angle before and after operation;and in PKP group, postoperative data has obviously improved than preoperative (P < 0.01), but there was no significant difference postoperative at 1 week, 12 weeks, 1 year (P > 0.05); there was no significant difference between two groups at 1 week, 12 weeks, 1 year after operation. CONCLUSION: Both the methods can obviously relieve pain and completely or partly recover daily life function in treating OVCF. But PKP has advantages of recovery of anterior height of responsibility vertebra and correction of Cobb angle, especially for serious compression.

Recombinant influenza H1, H5 and H9 hemagglutinins containing replaced H3 hemagglutinin transmembrane domain showed enhanced heterosubtypic protection in mice.

Influenza A viruses cause annual epidemics and irregular pandemics. A vaccine with heterosubtypic protection (hetero-protection) has been needed. In the present study, various influenza H1, H3, H5, and H9 hemagglutinin (HA) proteins were expressed in insect cells, and then mice were subcutaneously immunized with the expressed HA proteins, and challenged by influenza A viruses (A/Puerto Rico/8/1934 (H1N1) or A/chicken/Guangdong/96 (H9N2)). The results first showed that wild-type H3 hemagglutinin (HA) (H3-WT), but not a transmembrane domain (TM) mutant, had hetero-protection against both H1N1 and H9N2 with survival rates of 17% and 33% respectively, and that wild-type H1 (H1-WT), H5 (H5-WT) and H9 (H9-WT) had no hetero-protection against H1N1 or H9N2 except for H5-WT against H1N1 with a survival rate of 17%. Then the H3-WT TM replaced the TMs of H1-WT, H5-WT and H9-WT to generate recombinant H1-TM, H5-TM and H9-TM respectively, and whether the H3-WT TM-dependent hetero-protection could be transferred to these TM mutants was investigated. The results showed that the H3-WT TM-dependent hetero-protection was transferable. H1-TM against H9N2 and H9-TM against H1N1 were with survival rates of 33% and 17% respectively, and H5-TM against both H1N1 and H9N2 with survival rates of 50% and 17% respectively. Furthermore, higher dosage H5-TM scored 100% hetero-protection against H1N1. These results demonstrated that replacement of the TMs of non-H3 HAs with H3-WT TM could enhance their hetero-protection. These findings would help the development of future influenza vaccines against pandemics such as the recently appeared H7N9 infection.

Recombinant influenza A H3N2 viruses with mutations of HA transmembrane cysteines exhibited altered virological characteristics.

Influenza A H3N2 virus as the cause of 1968 pandemic has since been circulating in human and swine. Our earlier study has shown that mutations of one or two cysteines in the transmembrane domain of H3 hemagglutinin (HA) affected the thermal stability and fusion activity of recombinant HA proteins. Here, we report the successful generation of three recombinant H3N2 mutant viruses (C540S, C544L, and 2C/SL) with mutations of one or two transmembrane cysteines of HA in the background of A/swine/Guangdong/01/98 [H3N2] using reverse genetics, indicating that the mutated cysteines were not essential for virus assembly and growth. Further characterization revealed that recombinant H3N2 mutant viruses exhibited larger plaque sizes, increased growth rate in cells, enhanced fusion activity, reduced thermal and acidic resistances, and increased virulence in embryonated eggs. These results demonstrated that the transmembrane cysteines (C540 and C544) in H3 HA have profound effects on the virological features of H3N2 viruses.

Evidences for the existence of intermolecular disulfide-bonded oligomers in the H3 hemagglutinins expressed in insect cells.

The hemagglutinin (HA) protein as the predominant antigen, executes receptor binding and membrane fusion, which critically influence the virological characteristics of influenza viruses. The literature contained scattered data showing reduction-sensitive HA oligomers when HA proteins were analyzed under non-reducing conditions. However, whether the reduction-sensitive HA oligomers are inter-monomer disulfide-bonded has not been studied. Here, we showed: (1) the detection of ß-mercaptoethanol-sensitive H3 HA oligomers was not affected by the treatment of cells with iodoacetamide prior to cell solubilization; (2) H3 HA oligomers were present on cell surfaces; (3) H3 HA oligomers had higher density than monomers; and (4) mutation of all the five C-terminal cysteines completely abolished the formation of H3 HA oligomers. Furthermore, mutant HAs with mutations of TM cysteines, CT cysteines or all five cysteines had decreased thermal stability but increased fusion activity in comparison with wildtype HA. In conclusion, this study has presented enough evidence for the existence of inter-monomer S-S H3 HA oligomers formed by five C-terminal cysteines, and suggested that all five C-terminal cysteines exerted opposite effects on HA thermal stability and fusion activity.

Impact of the segment-specific region of the 3'-untranslated region of the influenza A virus PB1 segment on protein expression.

The 12 and 13 terminal nucleotides in the 3'- and 5'-untranslated regions (UTRs) of the influenza A virus genome, respectively, are important for the transcription of the viral RNA and the translation of mRNA. However, the functions of the segment-specific regions of the UTRs are not well known. We utilized an enhanced green fluorescent protein (eGFP) flanked at both ends by different UTRs (from the eight segments of H1N1 PR8/34) as a reporter gene to evaluate the effects of these UTRs on protein expression in vitro. The results showed that the protein expression levels of NP-eGFP, NS-eGFP, and HA-eGFP were higher than those of the other reporters and that the protein level of PB1-eGFP remained at a relatively low amount 48-h post-transfection. The results revealed that the UTRs of all segments differently affected the protein expression levels and that the effect of the UTRs of PB1 segment on protein expression was significant. The deletion of "UAAA" and "UAAACU" motifs in the PB1-3'-UTR significantly increased the protein expression level by 49.8 and 142.6%, respectively. This finding suggests that the "UAAACU" motif in the PB1-3'-UTR is at least partly responsible for the low protein expression level. By introducing the "UAAACU" motif into other 3'-UTRs (PA, NS, NP, and HA) at similar locations, the eGFP expression was reduced as expected by 56, 61, 22, and 22%, respectively. This result further confirmed that the "UAAACU" motif of the PB1-3'-UTR can inhibit protein expression. Our findings suggest that the segment-specific regions in the UTRs and not just the conserved regions of the UTRs play an important role in the viral protein expression. Additionally, the reported findings may also shed light on novel regulatory mechanism for the influenza A virus genome.

Mutations of two transmembrane cysteines of hemagglutinin (HA) from influenza A H3N2 virus affect HA thermal stability and fusion activity.

Influenza A H3N2 virus caused 1968 Hong Kong influenza pandemic, and has since been one of the most prevalent seasonal influenza viruses in global populations, representing a credible pandemic candidate in future. Previous studies have established that the hemagglutinin (HA) protein is the predominant antigen and executes receptor binding and membrane fusion. Homologous sequence analysis of all HA subtypes of influenza viruses revealed that two cysteine residues (540 and 544) are uniquely present in the transmembrane domain (TM) of HA proteins from all influenza A H3N2 viruses. However, the functions of these two cysteines have not been fully studied. Here, we generated three mutants (C540S, C544L, and 2C/SL) to investigate the effects of the two TM cysteines on the biological functions of H3 HA. We herein presented evidences that the mutations of one or two of the cysteines did not affect the proper expressions of HA proteins in cells, and more importantly all mutant H3 HAs showed decreased thermal stability but increased fusion activity in comparison with wildtype HA. Our results taken together demonstrated that the two TM cysteines are important for the biological functions of H3 HA proteins.

Analysis of conjugated linoleic acid and trans 18:1 isomers in synthetic and animal products.

The chemistry of conjugated fatty acids, specifically octadecadienoic acids (18:2; commonly referred to as conjugated linoleic acid, or CLA), has provided many challenges to lipid analysts because of their unique physical properties and the many possible positional and geometric isomers. After the acid-labile properties of CLAs during analytic procedures were overcome, it became evident that natural products, specifically dairy fats, contain one dominant (c9,t11-CLA), 3 intermediate (t7,c9-, t9,c11-, and t11,c13-CLA), and up to 20 more minor CLA isomers. The best analytic techniques to date include a combination of gas chromatography that uses 100-m highly polar capillary columns, silver ion-HPLC, and a combination of silver ion-thin-layer chromatography and gas chromatography to analyze the CLA and trans 18:1 isomers, because some of them serve as precursors of CLA in biological systems. These analytic techniques have assisted commercial suppliers to prepare pure CLA isomers and have permitted the evaluation of individual CLA isomers for their nutritional and biological activity in animal and human systems. It is increasingly evident that different CLA isomers have distinctly different physiologic and biochemical properties. These techniques are essential to evaluate dairy fats for their CLA content, to design experimental diets to increase the amount of CLA in dairy fats, and to determine the CLA profile in these CLA-enriched dairy fats. These improved techniques are used to evaluate the CLA profile in pork products from pigs fed different commercial CLA mixtures.

Methods for analysis of conjugated linoleic acids and trans-18:1 isomers in dairy fats by using a combination of gas chromatography, silver-ion thin-layer chromatography/gas chromatography, and silver-ion liquid chromatography.

Conjugated linoleic acids (CLA) are octadecadienoic acids (18:2) that have a conjugated double-bond system. Interest in these compounds has expanded since CLA were found to be associated with a number of physiological and pathological responses such as cancer, metastases, atherosclerosis, diabetes, immunity, and body fat/protein composition. The main sources of these conjugated fatty acids are dairy fats. Rumen bacteria convert polyunsaturated fatty acids, especially linoleic and linolenic acids, to CLA and numerous trans- containing mono- and diunsaturated fatty acids. It has been established that an additional route of CLA synthesis in ruminants and monogastric animals, including humans, occurs via delta9 desaturation of the trans-18:1 isomers. To date, a total of 6 positional CLA isomers have been found in dairy fats, each occurring in 4 geometric forms (cis,trans; trans,cis; cis,cis; and trans,trans) for a total of 24. All of these CLA isomers can be resolved only by a combination of gas chromatography (GC), using 100 m highly polar capillary columns, and silver-ion liquid chromatography, using 3 of these 25 cm columns in series. Complete analysis of all the trans-18:1 isomers requires prior isolation of trans monoenes by silver-ion thin-layer chromatography (TLC), followed by GC analysis using the same 100 m capillary columns operated at low temperatures starting from 120 degrees C. These analytical techniques are required to assess the purity of commercial CLA preparations, because their purity will affect the interpretation of any physiological and/or biochemical response obtained. Prior assessment of CLA preparations by TLC is also recommended to determine the presence of any other impurities. The availability of pure CLA isomers will permit the evaluation and analysis of individual CLA isomers for their nutritional and biological activity in model systems, animals, and humans. These techniques are also essential to evaluate dairy fats for their content of specific CLA isomers and to help design experimental diets to increase the level of the desired CLA isomers in dairy fats. These improved techniques are further required to evaluate the CLA profile in monogastric animals fed commercial CLA preparations for CLA enrichment of animal products. This is particularly important because absorption and metabolism will alter the ingested-CLA profile in the animal fed.

Evaluation of two GC columns (60-m SUPELCOWAX 10 and 100-m CP Sil 88) for analysis of milkfat with emphasis on CLA, 18:1, 18:2 and 18:3 isomers, and short- and long-chain FA.

Milkfat is a complex mixture of many diverse FA, some of which have demonstrated health benefits including anticancer properties. Attempts are under way to enrich milkfats with long-chain n-3 PUFA and CLA. It has been recommended that the analysis of these milkfats requires gas chromatography (GC) equipped with long, highly polar capillary columns. However, many analyses have been reported using CARBOWAX type (polyethylene glycol) capillary columns, such as SUPELCOWAX 10, even though the separation characteristics of many of the FA and their isomers present in milkfats have not been described in detail. This includes the isomers of CLA, cis- and trans-octadecenoic acid (18:1), linoleic acid (18:2n-6), and linolenic acid (18:3n-3), and the long-chain PUFA. On the other hand, the resolution of these FA and their isomers has been more fully described using the highly polar capillary columns, such as CP Sil 88 and SP2560 because of the improved resolution obtained using these polar columns. The present study was undertaken to characterize the separation of these FA present in milkfats using a 60-m SUPELCOWAX 10 column, to compare the results to those from a 100-m CP Sil 88 column, and to determine if these two columns could possibly serve to complement each other for the analysis of total milkfat. The advantages of the SUPELCOWAX 10 column were a better resolution of the short-chain saturated from their monounsaturated FA (MUFA) analogs, and a complete separation of the alpha-linolenic (18:3n-3) and eicosadecenoic acid (20:1) isomers. It also provided an alternative elution order of the linoleic (18:2n-6), 18:3n-3 and gamma-linolenic (18:3n-6) acid isomers. On the other hand, the CP Sil 88 column provided a better resolution of the CLA isomers, MUFA, the isolated cis and trans MUFA fractions, the PUFA, and many the 18:2n-6 and 18:3n-3 isomers. A complete analysis of milk lipids using the CP Sil 88 column required the prior separation of total FAME using silver ion-TLC. The results of the present study confirm that the 100-m highly polar capillary GC columns are mandatory for the analysis of milk lipids, and at best, the 60 m SUPELCOWAX 10 capillary column serves as a complementary GC column to provide different separations in certain regions based on its intermediate polarity.