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In the lead-up to the Truth Matters conference in October, we will be focusing our attention on the sufficiency, authority, and clarity of Scripture. One of our previous blog series, Looking for Truth in All the Wrong Places, strongly emphasizes those doctrines. The following entry from that series originally appeared on June 21, 2017. -ed.

If God is still speaking to His people today—particularly through mental impressions and premonitions—how can believers exercise discernment when it comes to interpreting and applying these divine messages? Put simply, how is following the private, subjective “leading” of the Lord any more reliable than gazing into a crystal ball?

As we saw last time, biblical discernment runs contrary to the kind of subjective mysticism many promote in the church today. Without any objective criteria, there is no means for determining truth from error. Such blithe subjectivity leaves people at the mercy of whatever mystical “voice” they’re listening to.

Upper Abdominal Distress

It is therefore ironic that advocates of mysticism inevitably treat discernment itself as if it were some kind of subjective, mystical ability. One author speaks of discernment as “a spiritual function,” by which he evidently means that discernment does not involve the intellect. ^[1] William DeArteaga, Quenching the Spirit (Lake Mary, FL: Creation House, 1992), 55. For a further discussion of DeArteaga’s work, see chapter 6 of Reckless Faith. In one of my earlier books I quoted Bill Hamon, one of the leading proponents of modern revelatory prophecy. Hamon’s recipe for discernment is a classic case of mystical anti-intellectualism. He believes prophecies can be properly evaluated only by people willing to set reason and logic aside:

I have sometimes heard people say, “I did not witness with that prophecy.” But after questioning them, I discovered that what they really meant was that the prophecy did not fit their theology, personal desires or goals, or their emotions reacted negatively to it. They failed to understand that we do not bear witness with the soul—the mind, emotions or will.

Our reasoning is in the mind, not the spirit. So our traditions, beliefs and strong opinions are not true witnesses to prophetic truth. The spirit reaction originates deep within our being. Many Christians describe the physical location of its corresponding sensation as the upper abdominal area.

A negative witness—with a message of “no,” “be careful” or “something’s not right”—usually manifests itself with a nervous, jumpy or uneasy feeling. There is a deep, almost unintelligible sensation that something is wrong. This sense can only be trusted when we are more in tune with our spirit than with our thoughts. If our thinking is causing these sensations, then it could be only a soulish reaction.

On the other hand, when God’s Spirit is bearing witness with our spirit that a prophetic word is right, is of God and is according to His will and purpose, then our spirit reacts with the fruit of the Holy Spirit. We have a deep, unexplainable peace and joy, a warm, loving feeling—or even a sense of our spirit jumping up and down with excitement. This sensation lets us know that the Holy Spirit is bearing witness with our spirit that everything is in order, even though we may not understand everything that is being said, or our soul may not be able to adjust immediately to all the thoughts being presented. ^[2] Bill Hamon, “How to Receive a Personal Prophecy,” Charisma (April 1991), 68 (emphasis added).

Notice that Hamon’s emphasis is entirely on feeling, while he derides the intellect, theology, reason, understanding, and by implication, true biblical wisdom. A reaction in the upper abdominal region is supposed to be a more reliable gauge of truth than all those things.

But that is superstition, not discernment. How your upper abdomen feels about a thing is certainly no measure of truth or falsehood. Neither is “a nervous, jumpy, or uneasy feeling” apart from any rational cause. “A deep, unexplainable peace and joy, a warm, loving feeling—or even a sense of [your] spirit jumping up and down with excitement” is no proof that a supposed prophecy is reliable. Those who practice this sort of “discernment” epitomize reckless faith.

And those who seek truth by analyzing inner feelings are likely to wind up with nothing but confusion.

Dueling Prophets

My editor once attended a service at the Anaheim Vineyard where two “prophets” gave contradictory prophecies. It happened in a Sunday morning worship service. When the congregational singing was over, John Wimber stepped to the platform. Before he could say anything, a young man in the congregation stood and began loudly to prophesy judgment against the leaders of the church. “Jerusalem! Jerusalem!” he began, echoing Luke 13:34, “you persecute My prophets and stone My messengers. My displeasure burns hot toward the leadership of this church for the way you have scorned My prophets and ignored My prophecies. . . .” and so on. The man evidently was disgruntled at the treatment he had received at the hands of church leaders, and this “prophecy” seemed to be his way of striking back. He prophesied in that manner for five minutes or more, earnestly calling the elders of the church to repentance. His entire message was in first person as if from God.

Immediately when he finished, before John Wimber could respond, another “prophet” from the other side of the congregation popped up and began to prophesy exactly the opposite message. This prophet began with a loose paraphrase of Jeremiah 29:11: “Oh, pastors and leaders of this church, I know My thoughts toward you—thoughts of mercy, and not of judgment. I have loved you with an everlasting love and have laid up for you a crown in heaven, My beloved. You have done according to all My good pleasure, and henceforth all men will rise up and call you blessed. . . .” etcetera, etcetera.

When the second man finished, a woman stood and sang a song, another person spoke in tongues, and one or two others quoted Bible verses or shared something brief. Then the service continued with Wimber making announcements. No reference was made to the two contradictory prophecies. No attempt was made to explain the dilemma or interpret either prophecy. Members of the congregation were simply left to draw their own conclusions about which, if either, of the two prophecies was correct.

That illustrates the impossible situation that arises when people are encouraged to voice their own subjective impressions as if they were divine prophecy. And it also reveals the predicament we are placed in if we must allow a sensation in our upper abdominal area to determine the truth or falsehood of a prophetic message.

Notice that both prophets’ messages echoed biblical terminology. Both of them were delivered with great conviction. Both of them employed first-person pronouns, as if God Himself were doing the speaking. Yet they flatly contradicted each other. They might both be false prophecies, but there is no way they could both be true. How were the people in the congregation supposed to determine which, if either, was correct? If they followed the gut-feeling approach, all the disgruntled people in the church undoubtedly opted for the first prophecy, believing they now had a word from the Lord to confirm their displeasure with their leaders.

The obvious fact is that once we stray into the realm of subjectivity, we have no way to determine what is really true.

(Adapted from Reckless Faith.)

Even though the central government issues strict regulation policies, it is the local governments’ discretion to adjust and enforce compliance.

Even though the central government issues strict regulation policies, it is the local governments’ discretion to adjust and enforce compliance.

Even though the central government issues strict regulation policies, it is the local governments’ discretion to adjust and enforce compliance.

Data from a large US cohort suggest systemic anticoagulation may confer a survival benefit in hospitalized patients without a spike in bleeding events.

Fatty liver involves ectopic lipid accumulation and dysregulated hepatic oxidative metabolism, which can progress to a state of elevated inflammation and fibrosis referred to as nonalcoholic steatohepatitis (NASH). The factors that control progression from simple steatosis to NASH are not fully known. Here, we tested the hypothesis that dietary vitamin E (VitE) supplementation would prevent NASH progression and associated metabolic alterations induced by a Western diet (WD). Hyperphagic melanocortin-4 receptor-deficient (MC4R^–/–) mice were fed chow, chow+VitE, WD, or WD+VitE starting at 8 or 20 weeks of age. All groups exhibited extensive hepatic steatosis by the end of the study (28 weeks of age). WD feeding exacerbated liver disease severity without inducing proportional changes in liver triglycerides. Eight weeks of WD accelerated liver pyruvate cycling, and 20 weeks of WD extensively upregulated liver glucose and oxidative metabolism assessed by ²H/¹³C flux analysis. VitE supplementation failed to reduce the histological features of NASH. Rather, WD+VitE increased the abundance and saturation of liver ceramides and accelerated metabolic flux dysregulation compared with 8 weeks of WD alone. In summary, VitE did not limit NASH pathogenesis in genetically obese mice, but instead increased some indicators of metabolic dysfunction.

A wide variety of intrinsic and extrinsic cues lead to cell death with unclear mechanisms. The infertility of some death mutants often hurdles the classical suppressor screens for death regulators. We have developed a transient RNA interference (RNAi)-based screen using a virus-induced gene silencing approach to understand diverse cell death pathways in Arabidopsis (Arabidopsis thaliana). One death pathway is due to the depletion of a MAP kinase (MAPK) cascade, consisting of MAPK kinase kinase 1 (MEKK1), MKK1/2, and MPK4, which depends on a nucleotide-binding site Leu-rich repeat (NLR) protein SUMM2. Silencing of MEKK1 by virus-induced gene silencing resembles the mekk1 mutant with autoimmunity and defense activation. The RNAi-based screen toward Arabidopsis T-DNA insertion lines identified SUMM2, MEKK2, and Calmodulin-binding receptor-like cytoplasmic kinase 3 (CRCK3) to be vital regulators of RNAi MEKK1-induced cell death, consistent with the reports of their requirement in the mekk1-mkk1/2-mpk4 death pathway. Similar with MEKK2, overexpression of CRCK3 caused dosage- and SUMM2-dependent cell death, and the transcripts of CRCK3 were up-regulated in mekk1, mkk1/2, and mpk4. MEKK2-induced cell death depends on CRCK3. Interestingly, CRCK3-induced cell death also depends on MEKK2, consistent with the biochemical data that MEKK2 complexes with CRCK3. Furthermore, the kinase activity of CRCK3 is essential, whereas the kinase activity of MEKK2 is dispensable, for triggering cell death. Our studies suggest that MEKK2 and CRCK3 exert concerted functions in the control of NLR SUMM2 activation and MEKK2 may play a structural role, rather than function as a kinase, in regulating CRCK3 protein stability.

Key Points

The TEAD family of transcription factors requires associating cofactors to induce gene expression. TEAD1 is known to activate the early promoter of human papillomavirus (HPV), but the precise mechanisms of TEAD1-mediated transactivation of the HPV promoter, including its relevant cofactors, remain unexplored. Here, we reveal that VGLL1, a TEAD-interacting cofactor, contributes to HPV early gene expression. Knockdown of VGLL1 and/or TEAD1 led to a decrease in viral early gene expression in human cervical keratinocytes and cervical cancer cell lines. We identified 11 TEAD1 target sites in the HPV16 long control region (LCR) by in vitro DNA pulldown assays; 8 of these sites contributed to the transcriptional activation of the early promoter in luciferase reporter assays. VGLL1 bound to the HPV16 LCR via its interaction with TEAD1 both in vitro and in vivo. Furthermore, introducing HPV16 and HPV18 whole genomes into primary human keratinocytes led to increased levels of VGLL1, due in part to the upregulation of TEADs. These results suggest that multiple VGLL1/TEAD1 complexes are recruited to the LCR to support the efficient transcription of HPV early genes.

IMPORTANCE Although a number of transcription factors have been reported to be involved in HPV gene expression, little is known about the cofactors that support HPV transcription. In this study, we demonstrate that the transcriptional cofactor VGLL1 plays a prominent role in HPV early gene expression, dependent on its association with the transcription factor TEAD1. Whereas TEAD1 is ubiquitously expressed in a variety of tissues, VGLL1 displays tissue-specific expression and is implicated in the development and differentiation of epithelial lineage tissues, where HPV gene expression occurs. Our results suggest that VGLL1 may contribute to the epithelial specificity of HPV gene expression, providing new insights into the mechanisms that regulate HPV infection. Further, VGLL1 is also critical for the growth of cervical cancer cells and may represent a novel therapeutic target for HPV-associated cancers.

The Epstein-Barr virus (EBV) causes human cancers, and epidemiological studies have shown that lytic replication is a risk factor for some of these tumors. This fits with the observation that EBV M81, which was isolated from a Chinese patient with nasopharyngeal carcinoma, induces potent virus production and increases the risk of genetic instability in infected B cells. To find out whether this property extends to viruses found in other parts of the world, we investigated 22 viruses isolated from Western patients. While one-third of the viruses hardly replicated, the remaining viruses showed variable levels of replication, with three isolates replicating at levels close to that of M81 in B cells. We cloned one strongly replicating virus into a bacterial artificial chromosome (BAC); the resulting recombinant virus (MSHJ) retained the properties of its nonrecombinant counterpart and showed similarities to M81, undergoing lytic replication in vitro and in vivo after 3 weeks of latency. In contrast, B cells infected with the nonreplicating Western B95-8 virus showed early but abortive replication accompanied by cytoplasmic BZLF1 expression. Sequencing confirmed that rMSHJ is a Western virus, being genetically much closer to B95-8 than to M81. Spontaneous replication in rM81- and rMSHJ-infected B cells was dependent on phosphorylated Btk and was inhibited by exposure to ibrutinib, opening the way to clinical intervention in patients with abnormal EBV replication. As rMSHJ contains the complete EBV genome and induces lytic replication in infected B cells, it is ideal to perform genetic analyses of all viral functions in Western strains and their associated diseases.

IMPORTANCE The Epstein-Barr virus (EBV) infects the majority of the world population but causes different diseases in different countries. Evidence that lytic replication, the process that leads to new virus progeny, is linked to cancer development is accumulating. Indeed, viruses such as M81 that were isolated from Far Eastern nasopharyngeal carcinomas replicate strongly in B cells. We show here that some viruses isolated from Western patients, including the MSHJ strain, share this property. Moreover, replication of both M81 and of MSHJ was sensitive to ibrutinib, a commonly used drug, thereby opening an opportunity for therapeutic intervention. Sequencing of MSHJ showed that this virus is quite distant from M81 and is much closer to nonreplicating Western viruses. We conclude that Western EBV strains are heterogeneous, with some viruses being able to replicate more strongly and therefore being potentially more pathogenic than others, and that the virus sequence information alone cannot predict this property.

Influenza D virus (IDV) was initially isolated in the United States in 2011. IDV is distributed worldwide and is one of the causative agents of the bovine respiratory disease complex (BRDC), which causes high morbidity and mortality in feedlot cattle. The molecular mechanisms of IDV pathogenicity are still unknown. Reverse genetics systems are vital tools not only for studying the biology of viruses, but also for use in applications such as recombinant vaccine viruses. Here, we report the establishment of a plasmid-based reverse genetics system for IDV. We first verified that the 3'-terminal nucleotide of each 7-segmented genomic RNA contained uracil (U), contrary to previous reports, and we were then able to successfully generate recombinant IDV by cotransfecting 7 plasmids containing these genomic RNAs along with 4 plasmids expressing polymerase proteins and nucleoprotein into human rectal tumor 18G (HRT-18G) cells. The recombinant virus had a growth deficit compared to the wild-type virus, and we determined the reason for this growth difference by examining the genomic RNA content of the viral particles. We found that the recombinant virus incorporated an unbalanced ratio of viral RNA segments into particles compared to that of the wild-type virus, and thus we adjusted the amount of each plasmid used in transfection to obtain a recombinant virus with the same replicative capacity as the wild-type virus. Our work here in establishing a reverse genetics system for IDV will have a broad range of applications, including uses in studies focused on better understanding IDV replication and pathogenicity, as well as in those contributing to the development of BRDC countermeasures.

IMPORTANCE The bovine respiratory disease complex (BRDC) causes high mortality and morbidity in cattle, causing economic losses worldwide. Influenza D virus (IDV) is considered to be a causative agent of the BRDC. Here, we developed a reverse genetics system that allows for the generation of IDV from cloned cDNAs and the introduction of mutations into the IDV genome. This reverse genetics system will become a powerful tool for use in studies related to understanding the molecular mechanisms of viral replication and pathogenicity and will also lead to the development of new countermeasures against the BRDC.

Recently, abundant evidence has clarified that long noncoding RNAs (lncRNAs) play an oncogenic or anticancer role in the tumorigenesis and development of diverse human cancers. Described as a crucial regulator in some cancers, MIR22HG has not yet been studied in laryngocarcinoma and therefore the underlying regulatory role of MIR22HG in laryngocarcinoma is worth detecting. In this study, MIR22HG expression in laryngocarcinoma cells was confirmed to be downregulated, and upregulated MIR22HG expression led to suppressive effects on laryngocarcinoma cell proliferation and migration. Molecular mechanism assays revealed that MIR22HG sponges miR-5000-3p in laryngocarcinoma cells. Besides, decreased expression of miR-5000-3p suppressed laryngocarcinoma cell proliferation and migration. Moreover, the FBXW7 gene was reported to be a downstream target gene of miR-5000-3p in laryngocarcinoma cells. More importantly, rescue assays verified that FBXW7 depletion or miR-5000-3p upregulation countervailed the repressive effects of MIR22HG overexpression on laryngocarcinoma progression. In addition, E2F6 was proved to be capable of inhibiting MIR22HG transcription in laryngocarcinoma cells. To sum up, E2F6-induced downregulation of MIR22HG promotes laryngocarcinoma progression through the miR-5000-3p/FBXW7 axis.

Flemming Dahlke, Magnus Lucassen, Ulf Bickmeyer, Sylke Wohlrab, Velmurugu Puvanendran, Atle Mortensen, Melissa Chierici, Hans-Otto Pörtner, and Daniela Storch

The vulnerability of fish embryos and larvae to environmental factors is often attributed to a lack of adult-like organ systems (gills) and thus insufficient homeostatic capacity. However, experimental data supporting this hypothesis are scarce. Here, by using Atlantic cod (Gadus morhua) as a model, the relationship between embryo vulnerability (to projected ocean acidification and warming) and homeostatic capacity was explored through parallel analyses of stage-specific mortality and in vitro activity and expression of major ion pumps (ATP-Synthase, Na⁺/K⁺-ATPase, H⁺-ATPase) and co-transporters (NBC1, NKCC1). Immunolocalization of these transporters was used to study ionocyte morphology in newly-hatched larvae. Treatment-related embryo mortality until hatch (+20% due to acidification and warming) occurred primarily during an early period (gastrulation) characterized by extremely low ion transport capacities. Thereafter, embryo mortality decreased in parallel with an exponential increase in activity and expression of all investigated ion transporters. Significant changes in transporter activity and expression in response to acidification (+15% activity) and warming (-30% expression) indicate some potential for short-term acclimatization, although likely associated with energetic trade-offs. Interestingly, whole-larvae enzyme capacities (supported by abundant epidermal ionocytes) reached levels similar to those previously measured in gill tissue of adult cod, suggesting that early-life stages without functional gills are better equipped in terms of ion homeostasis than previously thought. This study implies that the gastrulation period represents a critical transition from inherited (maternal) defenses to active homeostatic regulation, which facilitates enhanced resilience of later stages to environmental factors.

Interferon-regulated myxovirus resistance protein B (MxB) is an interferon-induced GTPase belonging to the dynamin superfamily. It inhibits infection with a wide range of different viruses, including HIV-1, by impairing viral DNA entry into the nucleus. Unlike the related antiviral GTPase MxA, MxB possesses an N-terminal region that contains a nuclear localization signal and is crucial for inhibiting HIV-1. Because MxB previously has been shown to reside in both the nuclear envelope and the cytoplasm, here we used bioinformatics and biochemical approaches to identify a nuclear export signal (NES) responsible for MxB's cytoplasmic location. Using the online computational tool LocNES (Locating Nuclear Export Signals or NESs), we identified five putative NES candidates in MxB and investigated whether their deletion caused nuclear localization of MxB. Our results revealed that none of the five deletion variants relocates to the nucleus, suggesting that these five predicted NES sequences do not confer NES activity. Interestingly, deletion of one sequence, encompassing amino acids 505–527, abrogated the anti-HIV-1 activity of MxB. Further mutation experiments disclosed that amino acids 515–519, and Pro-515 in particular, regulate MxB oligomerization and its binding to HIV-1 capsid, thereby playing an important role in MxB-mediated restriction of HIV-1 infection. In summary, our results indicate that none of the five predicted NES sequences in MxB appears to be required for its nuclear export. Our findings also reveal several residues in MxB, including Pro-515, critical for its oligomerization and anti-HIV-1 function.

The arrangement of functionally-related genes in operons is a fundamental element of how genetic information is organized in prokaryotes. This organization ensures coordinated gene expression by co-transcription. Often, however, alternative genetic responses to specific stress conditions demand the discoordination of operon expression. During cold temperature stress, accumulation of the gene encoding the sole Asp–Glu–Ala–Asp (DEAD)-box RNA helicase in Synechocystis sp. PCC 6803, crhR (slr0083), increases 15-fold. Here, we show that crhR is expressed from a dicistronic operon with the methylthiotransferase rimO/miaB (slr0082) gene, followed by rapid processing of the operon transcript into two monocistronic mRNAs. This cleavage event is required for and results in destabilization of the rimO transcript. Results from secondary structure modeling and analysis of RNase E cleavage of the rimO–crhR transcript in vitro suggested that CrhR plays a role in enhancing the rate of the processing in an auto-regulatory manner. Moreover, two putative small RNAs are generated from additional processing, degradation, or both of the rimO transcript. These results suggest a role for the bacterial RNA helicase CrhR in RNase E-dependent mRNA processing in Synechocystis and expand the known range of organisms possessing small RNAs derived from processing of mRNA transcripts.