High Enzyme Research Articles

Abstract 1807: Progestin enrichment of the cancer stem cell-like pool provides novel therapeutic targets for hormone-dependent human breast cancer

Abstract Clinical trials in post-menopausal women show that combined hormone replacement therapy (HRT), containing both estrogen (E) and a progestin (P) such as medroxyprogesterone acetate (MPA), is associated with an increased risk of breast cancer compared with E alone (JNCI, 2000, 92:328). We previously showed that both natural and synthetic P accelerate tumor development in vivo, a process that is blocked by anti-progestins (Can Res 2007; 67:9929). Furthermore, P also increases lymph node metastasis (Menopause 2010; 17:1040), leading us to hypothesize that P-induced tumor growth and metastasis may be mediated by an enrichment of the pool of cancer stem cell (CSC)-like cells. This increases the proliferative and metastatic potential of already existing tumors. CSCs are a small and highly tumorigenic cell population present in most tumors, which have self-renewal properties. In tumors of the breast, the CSC subpopulation has a phenotypic signature that is CD24 −/low, CD44 high and aldehyde dehydrogenase-positive (ALDH+) (Cell Stem Cell, 2007; 1:555). Using flow cytometry to measure CD44 density under a variety of experimental conditions we sought to determine whether P influences expression of CSC markers in P-responsive human breast tumors that grow in vivo. Following exposure of T47-D cells for 24 h with physiological levels (10 nM) of natural and synthetic P, CD44 protein expression was increased considerably. Compared with controls, induction of CD44 increased 10-fold in response to MPA (Control 5.9%, MPA 60.7%). Alternative steroid hormones failed to induce CD44 in tumor cells. This phenotypic shift was blocked by the anti-progestin RU-486, indicating that the process is mediated via progesterone receptors (PR). Cells treated with P formed mammospheres more easily than their non-treated counterparts, and a subset of the cell population with induced CD44 high also demonstrated high ALDH enzyme activity. RT-PCR analysis showed that MPA specifically induced CD44v3 and CD44v6 transcripts. Importantly, a variety of synthetic P similarly induced CD44 protein expression. Based on our observations we contend that exposure of breast cancer cells to synthetic P, such as MPA, may lead to an enrichment of the CSC-like pool, supporting the development of P-accelerated tumors in vivo. These findings also suggest that a novel means by which to combat P-dependent tumor growth might be through blockade of CD44 functions in the pool of stem cell-like cells. This could be achieved by immunotherapeutic means, using tissue selective antiprogestins, or through a combination approach involving both immunotherapy against CD44 and small molecule targeting of PR. Supported in part by COR award from the College of Veterinary Medicine, and by generous gifts from donors of Ellis Fischel Cancer Center, University of Missouri. Citation Format: Sandy Goyette, Benford Mafuvadze, Matthew T. Cook, Yayun Liang, Salman M. Hyder. Progestin enrichment of the cancer stem cell-like pool provides novel therapeutic targets for hormone-dependent human breast cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1807.

Feeding glycerol-enriched yeast culture improves lactation performance, energy status, and hepatic gluconeogenic enzyme expression of dairy cows during the transition period1

This study aimed to evaluate the effects of feeding glycerol-enriched yeast culture (GY) on feed intake, lactation performance, blood metabolites, and expression of some key hepatic gluconeogenic enzymes in dairy cows during the transition period. Forty-four multiparous transition Holstein cows were blocked by parity, previous 305-d mature equivalent milk yield, and expected calving date and randomly allocated to 4 dietary treatments: Control (no additive), 2 L/d of GY (75.8 g/L glycerol and 15.3 g/L yeast), 150 g/d of glycerol (G; 0.998 g/g glycerol), and 1 L/d of yeast culture (Y; 31.1 g/L yeast). All additives were top-dressed and hand mixed into the upper one-third of the total mixed ration in the morning from -14 to +28 d relative to calving. Results indicated that the DMI, NE intake, change of BCS, and milk yields were not affected by the treatments ( > 0.05). Supplementation of GY or Y increased milk fat percentages, milk protein percentages, and milk protein yields relative to the Control or G group ( < 0.05). Cows fed GY or G had higher glucose levels and lower β-hydroxybutyric acid (BHBA) and NEFA levels in plasma than cows fed the Control ( < 0.05) and had lower NEFA levels than cows fed Y ( < 0.05). On 14 d postpartum, cows fed GY or G had higher enzyme activities, mRNA, and protein expression of cytosolic phosphoenolpyruvate carboxykinase (PEPCK-C; < 0.05); higher enzyme activities ( < 0.05) and a tendency toward higher mRNA expression ( < 0.10) of glycerol kinase (GK); and a tendency toward higher enzyme activities of pyruvate carboxylase (PC) in the liver ( < 0.10) when compared with cows fed Control or Y. The enzyme activities, mRNA, and protein expression of PEPCK-C, PC, and GK did not differ between cows fed GY and G ( > 0.10). In conclusion, dietary GY or Y supplementation increased the milk fat and protein content of the cows in early lactation and GY or G supplementation improved the energy status as indicated by greater plasma glucose and lower plasma BHBA and NEFA concentrations and upregulated the hepatic gluconeogenic enzymes of dairy cows during the transition period. Feeding cows with a GY mixture in the peripartum period combined the effects of yeast on lactation performance and the effects of glycerol on energy status in dairy cows.

Characteristics of the Distribution of Acetylcholinesterase in the Posterolateral Nucleus of the Thalamus in Cats

Studies using histochemical detection of the enzyme acetylcholinesterase (AChE) showed that two-weekold cats (n = 4) had loci with higher enzyme activity in the lateral nucleus of the posterolateral complex of the thalamus and that these were not present in 14-week-old cats (n = 4). These loci were distributed opposite to similar loci in the medial nucleus of the posterolateral complex. A possible relationship between these AChE loci and the organization of thalamocortical and corticothalamic projections is discussed.

Fabrication of Ascorbic Acid Biosensor Based on Coupling Polyethylene Glycol Modified Silk Fibroin Membrane onto Glassy Carbon Electrode

Nowadays biosensors have been extensively used in a wide variety of applications especially in clinical works and food industry. In this work, a specific ascorbic acid (AA) biosensor was developed by immobilizing ascorbate oxidase (ASOD) on a polyethylene glycol (PEG) modified silk fibroin (SF) membrane then coupling to the glassy carbon electrode (GCE). The SF-PEG-ASOD membrane provided the highest enzyme activity in phosphate buffer at pH 5. As being the electrode, the SF-PEG-ASOD modified GCE displayed the highest response when it is operated under the condition of 0.40 mg/L of ASOD in phosphate buffer at pH 5. This biosensor provided both good linearity (r2 = 0.999 in the range of 1.0-10.0 mM) and sensitivity with short response time (26s). It also exhibited good anti-interference ability with the storage time of 5 days without changing its initial response.

Removing of Crude Oil From Polluted Areas Using the Isolated Fungi From Tehran Oil Refinery

ABSTRACTThe pollution of soil and the subsurface environment by crude oil spill and petroleum products spill is a major concern around the world. The aim of this research was to investigate the ability of fungi isolated from Tehran oil refinery area in removing crude oil and to evaluate their enzymatic activities. Plant root samples were collected from the polluted and control areas, and rhizospheral fungi were isolated and determined using the laboratory methods and taxonomic keys. Seven fungal species were isolated and then cultured in potato dextrose agar (PDA) media containing 0–15% (v/v) crude oil. Oil removal was determined after a one-month growth of fungal colonies and then compared with the control media. The results showed that the studied fungi were able to remove crude oil from the media. The highest removal efficiency was observed in Aspergillus sp. Total protein content and enzymatic activity (of peroxidase and catalase) increased with increasing crude oil pollution. The highest enzymatic activity was evaluated in Aspergillus sp. growing in media containing 15% petroleum and the lowest activity was found in non-polluted groups. Results showed that there is a direct correlation between oil-removing potency and enzymatic activity. Aspergillus sp. showed the highest enzyme activity and also the highest petroleum removal efficiency.

Stoichiometry of soil extracellular enzyme activity along a climatic transect in temperate grasslands of northern China

Ratios of specific carbon (C), nitrogen (N) and phosphorus (P) acquisition activities converged on 1:1:1 at a global scale and in tropical ecosystems. It is less clear if this pattern can be applied in temperate grasslands. The questions of whether the pattern remains stable across different soil depths and what the relative contributions are from the influence of climatic, edaphic abiotic and biotic factors on soil extracellular enzyme activity (EEA) stoichiometry remain uncertain. We measured potential activities of one C-acquiring enzyme (β-1, 4-glucosidase), two N-acquiring enzymes (β-N-acetylglucosaminidase and leucine aminopeptidase) and one organic P-acquiring enzyme (acid phosphatase) and major influential factors along a climatic transect in temperate grasslands of northern China during the growing season of 2013. We found lower enzyme C: N (0.47) and C: P (0.18) ratios and a higher enzyme N: P activity ratio (0.40) in 0–20 cm soil depth in temperate grasslands than in tropical soils (C: N, 1.83; C:P, 0.21; N: P, 0.13). The enzyme C: N and C: P ratios decreased with soil depth except for the enzyme C: N ratio in desert steppes. However, there were no significant differences in enzyme N: P ratio with soil depth. Among all the factors, soil total C, N and P contents accounted for the most variation in soil EEA and ecoenzymatic stoichiometry in 0–10 cm surface soil, which implied that soil EEA stoichiometry was largely controlled by soil nutrient stoichiometry. Moreover, edaphic factors had less influence on soil EEA stoichiometry in subsoil than in surface soil and edaphic abiotic factors had a larger effect on soil EEA stoichiometry than climatic and biotic factors. Our results suggest that soil extracellular enzyme activity ratios were not in homeostasis but resource dependent on soil and microbial biomass stoichiometry.

The Contribution of Attached Bacteria to Microcystis Bloom: Evidence From Field Investigation and Microcosm Experiment

ABSTRACTThe research of the relationships among free-living bacteria, attached bacteria and different algal species can provide valuable evidence for understanding the reason of algal bloom development under nutrient limitations. Significantly positive relationships between chlorophyll a and attached bacteria in Microcystis-bloom ponds and between chlorophyll a and free-living bacteria in other algal bloom ponds were observed. Microcosm experiments further confirmed the synchronized growth of Microcystis and attached bacteria as well as Pediastrum and free-living bacteria. Carbon and bacteria stimulation experiments suggested that extracellular organic carbon from Microcystis was more complex, inducing specific attached bacterial communities and markedly high extracellular enzymes, which can strongly fuelled Microcystis development. On the contrary, dissolved organic carbon from Pediastrum was simple and suitable for free-living bacteria, which can made a distinct effect on Pediastrum growth depending on the degree of nutrients uptake by free-living bacteria. Based on the above results, it can be proposed that the reason of Microcystis bloom breakout in Chinese shallow lakes should be attributed to the close nutrient complementary relationship between Microcystis and specific attached bacteria mediated by dissolved organic carbon and extracellular enzymes at the development of bloom, which might provide scientific support for the research of Microcystis bloom control.

A simple and rapid method for separation and isolation of marine algal species from naturally evolved populations

ABSTRACTTo improve the study of mixed microalgal populations, three naturally evolved marine microalgal cultures were subjected to a light crushing mechanical treatment using a silicon spatula coupled with zymolyase treatment at four concentrations: 5, 10, 20 and 25 U/ml, for 15, 30, 45 and 60 min before being observed under a microscope. The enzyme concentration of 20 U/ml after 45 min reduces the size of macroscopic microalgal aggregates and improves the microscopic observation of the different microalgal species comprising the population. There was no improvement using the higher enzyme concentration. This paper proposes a new approach to the study of naturally evolved microalgal populations which is useful for distinguishing the morphology of the different species present in the population and allowing for the identification by classical keys, and also to obtain a pure culture from an inoculum of mixed species by using a micromanipulator for cell counting.

Studies On hydrolysis of skimmed milk using immobilized β-galactosidase in a membrane reactor

In the present investigation, an attempt has been made to investigate the hydrolysis of skimmed milk in a batch membrane bioreactor using β-galactosidase immobilized on a 30 kDa PES membrane. The skimmed milk solutions of different concentrations(30 kg m-3 – 80 kg m-3)were deproteinated through 30 kDa and 5 kDa cross-flow ultrafiltration membrane modules successively and about 95%-97% lactose present in the feed was recovered in the permeate stream. The deproteinated milk permeates thus obtained were subjected to hydrolysis in a batch membrane bioreactor fitted with the enzyme-immobilized PES membrane. Immobilization of enzyme on the ultrafiltration membrane was performed by cross-linking using two different concentrations of glutaraldehyde (3%v/v and 4%v/v). It was observed that high percent retention ofenzyme activity(94.2%) and percent enzyme loading(98%) were attained when the immobilization was performed using 4%v/v glutaraldehyde. For an initial feed concentration of 70 kg m-3,the percentage lactose conversion achieved were 45.2% and 21.4% using β-galactosidase immobilized with 4%v/v and 3%v/v glutaraldehyde respectively. However, a significant percentage reduction in pure water fluxwas observed in case of enzyme-immobilized membranes (27.5% and 67.5% for immobilization carried out using 3% and 4%v/v glutaraldehyde respectively)from that obtained using the fresh membrane. The enzyme-immobilized membrane(cross-linked with 4%v/v glutaraldehyde) could be used for six successive runs with more than 90% retention in activity and less than 5% loss due to leaching. The unsteady state mass balance equation was developed in order to predict the concentration profile above the membrane surface.The model equation could also predict the permeate concentration well within 95% confidence interval

Reappraisal of the Significance of Inducible Nitric Oxide Synthase in Colorectal Cancer

Backgrounds: Inducible nitric oxide synthase (iNOS), which produces high levels of nitric oxide (NO), is overexpressed in activated macrophages and some cancer cells. Although iNOS was thought to be involved in promoting colorectal cancer, contradictory reports supporting its tumoricidal effect exist. Methods: We first examined the iNOS enzyme activity in colorectal cancer tissue and immunohistochemical expression of iNOS in cancer cells and tumor-infiltrating macrophages. Then, association of iNOS activity or its protein expression was analyzed in relation to various clinicopathological covariates. Results: Four groups of patients were classified based on their iNOS expression status. Univariate and multivariate analyses showed that group 1 patients (low iNOS in both types of cells) and group 4 patients (high iNOS in both types of cells) had a shorter disease-free survival. Patients with extremely high or low iNOS enzyme activity tended to have a lower disease-free survival rate (p = 0.059). Conclusion: Macrophage/stroma-derived NO negatively regulates colorectal cancer development when cancer cells express low levels of iNOS, but might synergistically contribute to tumor progression in the presence of high levels of cancer-cell derived NO. The dual effects of NO should be considered in the design of anti-iNOS/NO therapy for colorectal cancer patients.

Isolation of α-Amylase Producing Bacteria from Naini lake and Assessment of their Anti-Oxidant and anti-Fungal Properties

Bacterial enzymes play an important role in the industry. The objective of this study was to isolate and screen αamylase producing bacteria from water samples obtained from various regions of the Naini Lake, Uttarakhand, India. A total of 11 bacterial isolates were obtained on nutrient agar medium, which were then screened for the production of α-amylase. Antioxidant and anti-fungal property of the bacterial isolates which tested positive for α-amylase production was also assessed. Bacterial isolates NLWS 5 and NLWS 7 showed the highest enzyme production with a zone of 14mm and 10mm respectively. These two strains showed anti-oxidant as well as anti fungal activity which were then selected for identification using Bergey's Manual of Determinative Microbiology. It was found that the bacterial isolates which showed the highest amount of α-amylase production, anti-oxidant and anti-fungal activity belonged to Corynebacterium sp.

New Advancement on Cross-Linked Enzyme Aggregates within Magnetically-Separable Mesoporous Silica

Magnetically-separable enzyme system has been developed by adsorption, precipitation and cross-linking of enzymes in superparamagnetic hierarchically ordered mesoporous mesocellular silica (M-HMMS). The immobilization of xylanase within M-HMMS were compared between enzyme adsorption (EA), enzyme adsorption and cross-linking (EAC), and enzyme adsorption, precipitation and cross-linking (EAPC). EAPC includes higher enzyme activity immobilized within the matrix in comparison with the other methods. Furthermore, the immobilized enzyme is predicted to be prevented from leaching out of the matrix when exterior blow is being tested on the structure. Thus, the stability of the EAPC of this invention is anticipated to be maintained even after a long time passed since high enzyme activity compared with known method can be supported and immobilized within the matrix. Consequently, it is possible to improve performance of the enzymes by manipulating the preparation and operation condition.

Characterization of Crude Xylanase Produced by Edible Mushroom Pleurotus eryngii

Xylanase has been increasingly forthcoming in recent years because of their possible involvement in numerous industrial processes including bioconversion of lignocellulose derived sugars in to fuels, processing food and the paper and fiber industries. Edible mushrooms are emerging as important source of xylanolytic enzymes and this study has concentrated to produce and characterize xylanases by Pleurotus eryngii. The crude enzyme was characterized on the basis of various parameters such as incubation time, substrate specificity, substrate concentration, enzyme volume, buffer, pH, pH stability, temperature, temperature stability, and effect of various metal ions or compounds. The xylanase activity was noted maximum at 15 minutes of incubation time, 2.0% xylan and 0.5 ml enzyme volume. The highest enzyme activity was found at pH 4.5, whereas xylanase exhibited maximum stability in the range of pH 4.0 to 10.0. The maximum xylanase activity was noted at 60°C, while enzyme was most active and retains more than 40% activity at 90°C within 10 minutes of incubation. ZnCl2 (10mM) stimulated the xylanase activity as compare to other metal ions or compounds. It is concluded that Pleurotus eryngii is capable to produce pH stable and thermostable xylanase for industrial purposes.

Study the influence of nitrogen on rennin production by fungi Rhizomucor miehei using solid-state fermentation

The effect of different nitrogen resources on the biosynthesis of milk clotting enzyme by Rhizmucor miehei was studied under solid state fermentation using wheat bran as base medium. Urea, peptone, albumin, casein, yeast extract were added with different concentrations (1%-10%). The response parameters were the ratio of milk clotting activity (MC) to proteolytic activity (PA) and protein content. The highest enzyme yield was achieved with casein at a rate of 2% w/w followed by 2% yeast extract, 1% albumin, 1% peptone, and 1% urea with values 5.6, 4.9, 4.2, 4, 3 mg/mL, respectively. Maximum enzyme activity (MCA/PA) was 50.4, 44.1, 37.8, 36, 27 SU for casein, yeast extract, albumin, peptone, and urea, respectively.

Partial Purification and Characterization of Protease from &amp;lt;i&amp;gt;Abrus precatorius&amp;lt;/i&amp;gt; Linn. (Fabaceae) from Cameroon

Crude enzyme extracts were prepared from leaves and stems of Linn. (Fabaceae) from Cameroon under optimized conditions. Proteolytic enzymes were precipitated with ammonium sulfate at 35% (w/v) saturation and assayed for enzyme activity. The effects of temperature, pH, incubation time and substrate specificity were studied. SDS-PAGE was used to determine molecular weight of precipitated protease. Results indicated that proteolytic activity of crude extract was 35.20 U/ml compared to 51.03 U/ml of partial purified extract. The optimum enzyme activity was found to be at 40°C, while 50% of activity was maintained at 60°C after 60 min incubation. Partial purified crude extract exhibited two optimum pH (2.75 and 9.0). The highest enzyme activity towards Bovine Serum Albumine (25.9 U/ml) was noted. SDS-PAGE gels exhibited molecular weight between 40 - 60 KDa. This result confirms that partial purified extract of A. precatorius contains proteases and could be a promising source for proteolytic enzyme extraction.

Comparative study of the cholinesterase activities in tissues of the round goby Neogobius melanostomus (Gobiidae) from different locations of the Black and Azov seas

Cholinesterase activity has been assayed in the liver, muscles, and blood serum of the round goby Neogobius melanostomus caught in the coastal waters of the Black and Azov seas differing in the degree of anthropogenic impact. The highest enzyme activity in the fish individuals from both seas is observed in the liver and the lowest is observed in the blood serum. The fish caught in the most polluted (by municipal and agricultural wastewaters) sites of both seas display a decrease in cholinesterase activity. The utility of cholinesterase activity as a biomarker in monitoring of the marine areas affected by anthropogenic pollution is discussed.

굴과 굴 자숙액을 이용한 가수분해 조건의 최적화

Oyster is a nutritionally good food ingredient. Also, oyster is used to make source for taste and flavor. This study tried to investigate optimal condition of hydrolysis of oyster and oyster cooking drip for better amino acid content to make good taste and flavor. And then this study characterized hydrolysate of oyster and oyster cooking drip. Enzymes are Acalase, Flavourzyme, Neutrase, and Protamax. The optimal condition for the highest enzyme activity is given by the company. Under the best condition of each enzymes, they react with the homogenized oyster and oyster cooking drip for 0.5, 1.0, 1.5, 2, 4, 6 hr. The degree of oysters’ hydrolysis is 13.2±0.1%. But, in the case of using enzyme, the rate of hydrolysis sharply increased as time went on during 2 hr. After 8 hr, the rate is 36.9~40.5%. Protamax showed 27.4±0.4% of hydrolysis rate in 2 hr. And the degree of oyster cooking drop hydrolysis is 42.7±0.1%. The highest of hydrolysate concentration is 72.1±0.1% using protamax. In the case of oyster, it has a similar tendency of all enzymes. Otherwise, the hydrolysate of oyster cooking drip had a difference among the enzymes. Composition of free amino acid of hydrolysate using protamax was investigated how much time showed highest rate of hydrolysis to find best amino acid composition. Hydrolysis using Protamax during 6 hr is selected for best condition.

Highly Potent Saccharification of Arthrospira maxima Glycogen by Fungal Amylolytic Enzyme from Trichoderma Species J113

Hydrolytic enzymes from fungi have been widely used to convert complex plant-based substrates into more suitable fermentation substrates. Recently, photosynthetic microorganisms, particularly cyanobacteria, are garnering interest as an alternative to plant-based biomass for renewable energy production. Our goal was to identify a new source of enzymes with improved amylolytic efficiency in cyanobacterial glycogen hydrolysis. We isolated a new Trichoderma species J113 strain from the coastal terrains of Korea and determined that the fungus has a high amylolytic enzyme activity. We cultured the fungus on wheat bran to stimulate enzyme production, and the crude extract was subsequently purified through filtrations, precipitation, and chromatography. We observed that J113 enzyme consists of two putative major amylases: Ayt40 and Ayt70, that were determined as an α-amylase and a gluco-amylase, respectively. While these two amylases exhibited different pH and temperature requirements for optimum performance, collectively J113 enzyme showed the highest activity at pH 4 and 60 °C. In addition, we were able to drastically enhance the amylolytic capacity of Ayt70 gluco-amylase by 291 % with 5 mM Mn2+ amendment. Significantly, J113 enzyme converted 20 g/L (10 g total carbohydrate) of Arthrospira maxima to 8.3 g/L of reducing sugar with Mn2+ compared to only 5.1 g/L without Mn2+ in 240 min of reaction. Our study demonstrated that the newly isolated amylase extract from Trichoderma species J113 has the potential to be further optimized for efficient large-scale saccharification of algal glycogen for bioethanol production.

TERATOGENIC EFFECT OF HERBICIDE GLIFONOX IN RATTUS NORVEGICUS, WISTAR STRAIN

&lt;p&gt;Teratogenic effect of herbicide glyphosate-Roundup, sold under the name Glifotox on Wistar rats was evaluated. The biological material was treated intraperitoneally with glyphosate at concentrations of 100, 125, and 150 mg/kg from gestation day nine. Hysterectomy was performed on day 18 of gestation, and the uterine horns where the embryos were located, in addition to recording the percentage of malformed embryos by modifying the method of Wilson were observed. The liver was removed and quantified by spectrophotometry with transaminase activity showed higher concentrations malformation rate and higher enzyme activity was 125 mg/kg, below is the average of 100 mg/kg and higher concentrations such as 150 mg/kg a large number of resorptions was obtained. It is concluded that glyphosate is toxic affecting the liver and liver enzymes involved in the formation of amino acids also produce delay in embryonic development.&lt;/p&gt;

Acute Fatty Liver of Pregnancy Complicated by Quadruplets

Poster Presentation Background The diagnosis of acute fatty liver of pregnancy (AFLP) prior to surgical incision is critical. Case A 26‐year‐old G2P1 woman pregnant with quadruplets at 32 weeks gestation presented to triage with general malaise, pain in the right thigh, and feeling unwell. The woman had normal range blood pressures and no complaint of visual changes, epigastric pain, or jaundiced appearance. She was admitted for observation to rule out deep vein thrombosis (DVT). Routine labs were ordered with a comprehensive metabolic panel that indicated high enzymes, low platelets, and low hemoglobin and hematocrit (H&H). The decision to proceed with delivery was made due to diagnosis of HELLP syndrome. Clotting studies and a disseminated intravascular coagulation (DIC) panel were ordered and drawn at this time. The woman was sent to the operating room (OR) for delivery and prepped for an epidural. The diagnosis of AFLP was made in the OR. The procedure was stopped prior to epidural placement, and the woman was moved back to the obstetric intensive care unit for correction of coagulopathy. Consultation was initiated with transfusion medicine and transfusion of 12 units apheresis platelets, two units of packed red blood cells (PRBC), 16 pooled cryoprecipitated anti‐hemophilic factor (cyro), two jumbo fresh frozen plasma (FFP) units, and six liquid plasma units was ordered. We increased fibrinogen from 60 to 347 during the course of 6 hours, and the decision was made to move the woman to the OR and proceed with the cesarean with an intraoperative transfusion of three apheresis platelets, two units of PRBCs, six pooled cyros, and two jumbo FFP units. The delivery of the quadruplets and delivery of placenta were successful with good hemostasis. The woman experienced a total estimated blood loss of 1400 ml. After birth, the quadruplets were transported to the NICU, and the women was ventilated and placed in the obstetric intensive care unit. Careful evaluation for renal failure, hepatic encephalopathy, and/or other sequelae of AFLP was ongoing. The woman was extubated 24 hours postoperatively with stable renal function and urine output. Conclusion On postoperative day 2, the woman began to experience agitation, deterioration in mental status, and decreased ability to communicate. Fibrinogen decreased from 327 to 225, international normalized ratio (INR) increased from 2.2 to 2.5, ammonia increased from 33 to 51, and creatinine was 2.60. A care team including renal specialists, critical care providers, maternal/fetal medicine and obstetric intensive care unit nurses agreed to transfer the woman to a long‐term acute care facility with an in‐house live transplant team for ongoing support and care. The diagnosis of AFLP prior to surgical incision proved critical to this woman's survival. The multidisciplinary care team worked together to provide optimal care and prevented what could have been a dire outcome.