Implicit Value Updating Explains Transitive Inference Performance: The Betasort Model

Transitive inference (the ability to infer that B > D given that B > C and C > D) is a widespread characteristic of serial learning, observed in dozens of species. Despite these robust behavioral effects, reinforcement learning models reliant on reward prediction error or associative strength routinely fail to perform these inferences. We propose an algorithm called betasort, inspired by cognitive processes, which performs transitive inference at low computational cost. This is accomplished by (1) representing stimulus positions along a unit span using beta distributions, (2) treating positive and negative feedback asymmetrically, and (3) updating the position of every stimulus during every trial, whether that stimulus was visible or not. Performance was compared for rhesus macaques, humans, and the betasort algorithm, as well as Q-learning, an established reward-prediction error (RPE) model. Of these, only Q-learning failed to respond above chance during critical test trials. Betasort’s success (when compared to RPE models) and its computational efficiency (when compared to full Markov decision process implementations) suggests that the study of reinforcement learning in organisms will be best served by a feature-driven approach to comparing formal models.     

Nonverbal transitive inference: Effects of task and awareness on human performance

We studied human nonverbal transitive inference in two paradigms: with choice stimuli orderable along a physical dimension and with non-orderable choice stimuli. We taught 96 participants to discriminate four overlapping pairs of choice stimuli: A+ B−, B+ C−, C+ D−, and D+ E−. Half of the participants were provided with post-choice visual feedback stimuli which were orderable by size; the other half were not provided with orderable feedback stimuli. In later testing, we presented novel choice pairs: BD, AC, AD, AE, BE, and CE. We found that transitive responding depended on task awareness for all participants. Additionally, participants given ordered feedback showed clearer task awareness and stronger transitive responding than did participants not given ordered feedback. Associative models ( [Wynne, 1995] and [Siemann and Delius, 1998]) failed to predict the increase in transitive responding with increasing awareness. These results suggest that ordered and non-ordered transitive inference tasks support different patterns of performance.     

The use of gel chromatography to study rapid equilibria of the type A + B forms and is formed from C + D

A frontal gel chromatographic procedure is illustrated whereby the equilibrium constant for hybridization equilibria (A + B ? C + D) may be obtained provided the relative elution volume situation V_A = V_C <V_B = V_D may be realized. The electron transfer between reduced cytochrome c and ferricyanide is used as a model interaction, with Sephadex G-25 as the gel medium.     

Transverse patterning in pigeons

The ability of pigeons to form configural stimulus representations was assessed in two operant discrimination experiments. In Experiment 1 the transverse patterning problem of Spence (1952) was trained. In Phase 1, subjects had to choose stimulus A on A + B — trials; B + C — trials were added in Phase 2. In Phase 3, the first two pairs were combined with C + A — trials. The success of the subjects was simulated by a model assuming that elemental and configural stimulus representations coexist in a stable proportion, even in the phases of the experiment which do not require configural stimulus representations for successful solution. Experiment 2 replicated the first two phases of Experiment 1, but trained A + C — in Phase 3. Comparison of the results of this experiment with simulations of the model showed that elemental and configural stimulus representations coexisted in similar proportions as in Experiment 1, even though they were not necessary for successful task solution.     

Synthesis,insecticidal activities and SAR of novel phthalamides targeting calcium channel

In order to find novel and environmental friendly insecticides targeting the ryanodine receptor, three series of novel phthalamides containing heptafluoroisopropyl group, low fluorine atoms group and non-fluorine group were designed and synthesized. 35 novel structures of three series were obtained. Insecticidal activities of title compounds against oriental armyworm (Mythimna separata) and diamondback moth (Plutella xylostella) indicated that most of title compounds showed moderate to high activities at the tested concentration. The structure–activity relationship (SAR) was discussed in detail. During synthesizing title compounds B₈, C₇, D₁, D₉ and D₁₂, their corresponding positional isomers (B₈′, C₇′, D₁′, D₉′ and D₁₂′) were afforded, and their structures were confirmed by 2D NMR. The calcium-imaging technique was also applied to investigate the effects of compounds B₂, B₁₀, C₄ and C₅ on the intracellular calcium ion concentration ([Ca²⁺]_i), which indicated that they released stored calcium ions from endoplasmic reticulum, which denoted that some compounds are potential modulators of the insect ryanodine receptor (RyR).     

Cognitive representation in transitive inference: A comparison of four corvid species

During operant transitive inference experiments, subjects are trained on adjacent stimulus pairs in an implicit linear hierarchy in which responses to higher ranked stimuli are rewarded. Two contrasting forms of cognitive representation are often used to explain resulting choice behavior. Associative representation is based on memory for the reward history of each stimulus. Relational representation depends on memory for the context in which stimuli have been presented. Natural history characteristics that require accurate configural memory, such as social complexity or reliance on cached food, should tend to promote greater use of relational representation. To test this hypothesis, four corvid species with contrasting natural histories were trained on the transitive inference task: pinyon jays, Gymnorhinus cyanocephalus; Clark's nutcrackers, Nucifraga columbiana; azure-winged magpies, Cyanopica cyanus; and western scrub jays, Aphelocoma californica. A simplified computer model of associative representation displayed a characteristic pattern of accuracy as a function of position in the hierarchy. Analysis of the deviation of each subject's performance from this predicted pattern yielded an index of reliance on relational representation. Regression of index scores against rankings of social complexity and caching reliance indicated that both traits were significantly and independently associated with greater use of relational representation.     

Polymorphism of DNA double helices

Native DNA duplexes in fibers exist usually in one of three well-known (A, B and C) forms depending on relative humidity, type of cations and the amount of retained salt. To determine the precise influence of these factors and the effect of base composition, as well as base sequence, on DNA secondary structure, X-ray diffraction methods have been used to study all four synthetic DNA duplexes with repeated dinucleotide sequences, eight of the 12 with repeated trinucleotide sequences and seven analogues in which guanine was replaced with hypoxanthine. The results indicate that there are at least six additional allomorphs denoted by B′, C′, C″, D, E and S.The B′ form (h = 0.329 nm) observed for poly(dA) · poly(dT), poly(dI) · poly(dC) and poly[d(A-I)] · poly[d(C-T)] is a minor variant of the traditional B form (h = 0.338 nm) of native DNA. The two C-like forms C′ for poly[d(A-G-C)] · poly-[d(G-C-T)] and poly[d(G-G-T)] · poly[d(A-C-C)] and C″ for poly[d(A-G)] · poly-[d(C-T)] have, respectively, 9₁ and 9₂ symmetries which reflect repetition of trinucleotide and dinucleotide sequences, respectively. Although isocompositional with poly(dA) · poly(dT), the existence of the rather different D form (8₁) for poly[d(A-T)] · poly[d(A-T)] or for poly[d(A-A-T)] · poly[d(A-T-T)] is a clear demonstration of the sequence effect. The I · C pair generally mimics an A · T pair, but poly[d(I-I-T)] · poly[d(A-C-C)] provides a new (E) form with approximately 15₂ screw symmetry and with 〈h〉 = 0.325 nm and 〈t〉 = 48 dg per nucleotide. The S form (6₅) observed for poly[d(G-C)] · poly[d(G-C)] and poly[d(A-C)] · poly[d(G-T)] is an unusual left-handed polydinucleotide helix and is accessible to any alternating purine-pyrimidine sequence. In it the two nucleotides have quite different conformations and involve syn purine and anti pyrimidine nucleosides.     

Control of Endosperm Proteins in TRITICUM AESTIVUM (Var. Chinese Spring) and AEGILOPS UMBELLULATA by Homoeologous Group 1 Chromosomes

The genetic control of major wheat endosperm proteins by homoeologous group 1 chromosomes has been studied by two-dimensional polyacrylamide gel electrophoresis. The control of at least 15 distinct protein subunits or groups of protein subunits has been allocated to chromosomes 1A, 1B and 1D of Chinese Spring wheat from the analysis of grains of aneuploid genotypes. In addition, six protein subunits have been shown to be controlled by chromosome 1C^u of the related species, Aegilops umbellulata, from studies of wheat lines carrying disomic substitutions of 1C^u chromosomes. On the basis of protein subunit patterns, chromosome 1C^u is more closely related to chromosome 1D of wheat than to chromosomes 1A or 1B.     

DNA synthesis during the division cycle of three substrains of Escherichia coliBr

The relationship between chromosome replication and the bacterial division cycle has been examined in three substrains of Escherichia coli$\text{B}\text{r}$ obtained from different sources and designated $\text{B}\text{r}$ A, $\text{B}\text{r}$ F and $\text{B}\text{r}$ K. At growth rates greater than 1.0 doubling per hour (μ > 1.0), the time for a round of chromosome replication (C) was 42 minutes in all three substrains, but the time between the end of a round and cell division (D) was 22 minutes in $\text{B}\text{r}$ A, 16 minutes in $\text{B}\text{r}$ F and 14 minutes in $\text{B}\text{r}$ K. At slower growth rates C and D increased, but to significantly different extents in the three substrains. When μ = 0.5, C and D were approximately 80 and 40 minutes in $\text{B}\text{r}$ A, 60 and 20 minutes in $\text{B}\text{r}$ F, and 70 and 20 minutes in $\text{B}\text{r}$ K.As a consequence of the lengths of the C and D periods in the three stocks of E. coli$\text{B}\text{r}$, the patterns of chromosome replication during the division cycle differed. The most obvious difference was that E. coli$\text{B}\text{r}$ F and E. coli$\text{B}\text{r}$ K possessed periods devoid of DNA synthesis at both the beginning and the end of the division cycle during slow growth, whereas E. coli$\text{B}\text{r}$ A contained only one period devoid of DNA synthesis at the end of the cycle.     

Concentration equations for the diffusion of a reversibly reacting substance in gel-like media

The concentration of a diffusible substanceA(x, t) in a semi-infinite geometry is studied for the set of reversible reactionsA+B _i ?C _i ;i=1...n, whereB _i andC _i are assumed to be associated with non-diffusible biological structures. Assuming chemical equilibrium prevails throughout for each reaction, it is shown that a single uncoupled partial differential equation is sufficient to specifyA(x, t) and indirectlyB _i (x, t) andC _i (x, t) as well: $$\left[ {1 + \sum\limits_i {\frac{{K_i \beta _i }}{{\left( {1 + K_i A} \right)^2 }}} } \right]\frac{{\partial A}}{{\partial t}} = D_A \frac{{\partial ^2 A}}{{\partial x^2 }}$$ whereK _i is the chemical equilibrium constant of theith reaction, β₁ is concentration of binding sites of theith species (i.e.B _i+C _i) andD _A is the usual diffusion constant forA. Numerical solutions for boundary conditions amenable to the Boltzman transformation are presented and the range of parameters established over which the uniqueness and convergence of the solutions can be proven.     

Photoneutron-induced damage reduction for cardiac implantable electronic devices using neutron-shielding sheets in high-energy X-ray radiotherapy: A phantom study

PurposeTo evaluate damage reduction in cardiac implantable electronic devices (CIEDs) caused by photoneutrons in high-energy X-ray radiotherapy using a neutron-shielding sheet (NSS).MethodsThe NSS consists of a bolus with a thickness of 1 or 2 cm (Bls1 or Bls2) as a moderator and several absorbers (20%, 50%, or 80% B₄C silicone sheet [B₄C20, B₄C50, or B₄C80] or a 40% LiF silicone sheet [LiF40]). First, a linear accelerator (LINAC) with a water-equivalent phantom was modeled in the simulation and measured experimentally. Several NSSs were placed on the phantom, a Eu:LiCaAlF₆ scintillator was placed between the phantom and the NSS, and X-rays were irradiated. The relative counts (Cr = counts when placing the NSS or Bls2) were compared between the experiment and simulation. Second, CIED damage was evaluated in the simulation. The relative damage (Dr = damage when placing or not placing the NSS) was compared among all the NSSs. In addition, the γ-ray and leaking X-ray dose from B₄C was measured using a dosimetric film. After determining the optimal NSS combination, Dr value analysis was performed by changing the length of one side and the thickness.ResultsThe Cr values of the simulation and experiment agreed within a 30% percentage difference, except for Bare or LiF40-only. The Dr value was reduced by 43% when Bls2 + B₄C80 was applied. The photon dose was less than 5 cGy/1500 MU. The Dr values were smaller for the smaller lengths of one side of B₄C80 and decreased as the M-layer thickness increased.ConclusionsThe CIED damage induced by photoneutrons generated by a LINAC was effectively reduced by applying the optimal NSS.     

Vanadyl(IV) conalbumin. II. Mixed metal and anion binding studies

Electron paramagnetic resonance (epr) studies demonstrate that at low levels of conalbumin (CA) saturation with Fe³⁺ or VO²⁺, a ph-dependent preference of the metal exists for different protein binding-site configurations,A, B, and C. The vanadyl ion epr spectra of mixed VO²⁺, Fe³⁺-conalbumin in which Fe³⁺ is preferentially bound to the N- or C-terminal binding site are consistent with all three configurations being formed at both metal sites. At high pH the spectra suggest interaction between binding sites. In the absence of HCO₃^?, VO²⁺ is bound almost exclusively in B configuration; a full binding capacity of 2 VO²⁺ per CA is retained. Stoichiometric amounts of HCO₃^? convert the epr spectrum from B to an A, B, C type. Addition of oxalate to bicarbonate-free preparations converts the B spectrum to an A′, B, C′ type where the B resonances have lost intensity to the A′ and C′ resonances but have not changed position. The data suggest that configuration B is anion independent and that only one equivalent of binding sites at pH 9 responds to the presence of HCO₃^1? or oxalate by changing configuration but not metal binding capability. The form of the bound anion may be HCO₃^? rather than CO₃^2?. The formation rate of the colored ferric conalbumin complex by oxidizing Fe²⁺ to Fe³⁺ in limited HCO₃^? at pH 9 is also consistent with one equivalent of sites having different anion requirements than the remaining sites. Increased NaCl or NaClO₄ concentration or substitution of D₂O for water as solvent affect the environment of bound VO²⁺, but the mechanisms of action are unknown.     

Uptake,incorporation and calcium-ionophore-stimulated mobilization of arachidonic,eicosapentaenoic and docosahexaenoic acids by leucocytes of the rainbow trout,Salmo gairdneri

Rainbow trout leucocytes contain high levels of neutral lipid (about 70% of total lipid on a wt% basis) consisting of mostly triacylglycerol, free sterols and sterol esters (25%, 15% and 52% of neutral lipid, respectively). The phospholipids, separated by thin-layer chromatography, consisted predominantly of phosphatidylcholine, phosphatidylethanolamine and phosphatidylserine, each present at about 30% of the total phospholipid. Radiolabelling of the leucocytes for 1 h with 1 μCi (approx. 6 μM) [1−¹⁴C]20:4(n−6), [1−¹⁴C]20:5(n−3) or [1−¹⁴C]22:6(n−3) each gave similar uptake values (approx. 1 · 10⁵ cpm/10⁷ leucocytes). The incorporation into total phospholipids was highest for 22:6(n−3) and lowest for 20:4(n−6). A higher percentage of radiolabel from [1−¹⁴C]22:6(n − 3) was found incorporated into phosphatidylcholine and phosphatidylethanolamine as compared to that from [1−¹⁴C]20:4(n − 6) and [1−^14C]20:5(n−3), while the reverse situation was found with phosphatidylinositol and phosphatidylserine. The relative rates of incorporation into the different phospholipid classes for all three fatty acids were in the order phosphatidylinositol > sphingomyelin > diphosphatidylglycerol > phosphatidylcholine > phosphatidylethanolamine > phosphatidylserine. Calcium ionophore-challenge did not significantly alter the pattern of phospholipid radiolabel. Ionophore-challenge released large amounts of radiolabel, much of which was recovered after high-performance liquid chromatographic separation as free fatty acid/monohydroxy fatty acids, although only approx. 0.3% was recovered in leukotriene B₄ and leukotriene B₅ for the [1−¹⁴C]20:4(n−6) and [1−¹⁴C]20:5(n−3) labelled leucocytes, respectively. Other lipoxygenase products were also radiolabelled and tentatively identified as 20-carboxy-LTB₄, 20-hydroxy-LTB₄, 6-trans-LTB₄, 6-trans-12-epi-LTB₄, 6-trans-8-cis-12-epi-LTB₄ and the corresponding LTB₅ structures. No ‘6-series’ leukotrienes were produced from [1−¹⁴C]22:6(n−3), nor was there any evidence for the synthesis of ‘5-series’ leukotrienes via retroconversion of 22:6(n−3) to 20:5(n−3). This latter finding shows that, despite the preponderance of 22:6(n−3) in the membranes of trout leucocytes, this fatty acid is not a substrate for leukotriene generation.     

Computer simulations of mass transport for nonidentical interacting molecules.

We have developed a system for simulating the mass transport properties of interacting nonidentical macromolecules where the association is of the form A + B ? C, C + B ? D. Our simulation programs operate in a minicomputer (PDP 1104) with 16K of core and provide results identical to methods previously usable only with large computers. We use a rectangular approximation for the centrifuge cell which greatly simplifies calculation, although it introduces a few percent error into any attempt at quantitative fitting of actual data. The program as written is directly applicable to gel chromatography, simply by substitution of flow for centrifugal field and dispersion for diffusion. Simulations of centrifuge results have been compared with experimental results for two systems which have been proposed to fit the association pattern described—nitrogenase components and an antigen-antibody interaction. In both cases the results of our simulations suggest that the accepted interpretation of the experimental results may need to be modified. For the antigen-antibody interaction, the presence of multivalent higher order complexes apparently is required to explain the centrifuge results. For nitrogenase, one cannot readily distinguish the case of association to form both 1:1 and 1:2 molar complexes from that of formation of only the 1:1 complex on the basis of the published data. Criteria for making such a discrimination are discussed.     

Evolution of the three overlapping gene systems in G4 and phi X174.

Bacteriophage G4 has the same $\text{A}\text{B}$ and $\text{D}\text{E}$ overlapping gene systems as φX174 and together with the A and $\text{C}\text{K}$ overlapping gene system (Shaw et al., 1978), 7 of the 11 G4 and φX174 genes are involved in overlaps. The nucleotide differences between G4 and φX174 in the overlapping portions of the A, C and D genes are 23%, 27% and 21%, respectively, compared with 32%, 36% and 34% in the non-overlapping portions of the same genes. The amino acid differences between the G4 and φX174 overlapping B, K and E proteins, are 44%, 39% and 44%, respectively, compared with 28%, 26% and 16% in the regions of genes A, A and C, and D which contain genes B, K and E. These results suggest that the nucleotide sequences of overlapping genes evolve at almost the same rate as in non-overlapping genes, and that this is made possible by a lower amino acid sequence stringency of one of the pairs of proteins. The overlapping $\text{D}\text{E}$ and A and $\text{C}\text{K}$ gene systems may have originated by taking advantage of a high incidence of T nucleotides in the second codon position to produce a hydrophobic protein and the $\text{A}\text{B}$ gene system may have evolved by read-through of the A gene into the B gene. From the nucleotide sequences, other overlapping genes appear to be possible in these bacteriophages.     

An analysis of the influence of age and school-attendance status on the spread of variola minor.

Definitions are proposed for the independent and joint contributions that the chemical groups A and B make to the free energy of association of the ligand A?B with a receptor. The definitions are independent of the choice of the standard state and are consistent with the basic thermodynamic cycle relating the association of the ligands A?B, A?Y and X?B to the receptor Rappaport 1976. The basic idea is the use of the excess free energy of association of the ligand A?Y over the free energy of association of the reference ligand X?Y as the measure of the “independent” contribution of the group A to the binding. This definition allows the free energy of association of the ligand A?B to be written as the sum of the independent contributions of the groups A and B, their joint contribution, and an invariant free energy of association of the reference ligand with any receptor. With the appropriate definition of the receptor-reference ligand complex, water can be chosen as the reference ligand. Using ΔG(A?OH)?AG(HOH), ΔG(H?B?H)?ΔG(HOH) and ΔG(HO?C)?ΔG(HOH) as the definitions of the “independent” contributions of the chemical groups A, B and C to the binding of the ligand A?B?C, the joint contribution of the groups A and C to the binding is ΔG(A?B?C) ? ΔG(A?B?H) ? ΔG(H-B-C) + ΔG(H?B?H).     

The Leptin,Dopamine and Serotonin Receptors in Hypothalamic POMC-Neurons of Normal and Obese Rodents

The pro-opiomelanocortin (POMC)-expressing neurons of the hypothalamic arcuate nucleus (ARC) are involved in the control of food intake and metabolic processes. It is assumed that, in addition to leptin, the activity of these neurons is regulated by serotonin and dopamine, but only subtype 2C serotonin receptors (5-HT_2CR) was identified earlier on the POMC-neurons. The aim of this work was a comparative study of the localization and number of leptin receptors (LepR), types 1 and 2 dopamine receptors (D₁R, D₂R), 5-HT_1BR and 5-HT_2CR on the POMC-neurons and the expression of the genes encoding them in the ARC of the normal and diet-induced obese (DIO) rodents and the agouti mice (A ^y /a) with the melanocortin obesity. As shown by immunohistochemistry (IHC), all the studied receptors were located on the POMC-immunopositive neurons, and their IHC-content was in agreement with the expression of their genes. In DIO rats the number of D₁R and D₂R in the POMC-neurons and their expression in the ARC were reduced. In DIO mice the number of D₁R and D₂R did not change, while the number of LepR and 5-HT_2CR was increased, although to a small extent. In the POMC-neurons of agouti mice the number of LepR, D₂R, 5-HT_1BR and 5-HT_2CR was increased, and the D₁R number was reduced. Thus, our data demonstrates for the first time the localization of different types of the serotonin and dopamine receptors on the POMC-neurons and a specific pattern of the changes of their number and expression in the DIO and melanocortin obesity.     

Temperature and pressure dependent equilibrium studies of 1, 4, 8, 11-tetramethyl-1, 4, 8, 11-tetraazacyclotetradecane nickel(ii) in coordinating solvents

In coordinating solvents, the complex 1, 4, 8, 11- tetramethyl-1, 4, 8, 11-tetraazacyclotetradecane nickel(II) bisperchlorate exists as an equilibrium mixture involving four coordinate R,S,R,S-[Ni(tmc)]²⁺ and five coordinate R,S,R,S-[Ni(tmc)(solvent)]²⁺ species. Spectrophotometric measurements of this equilibrium in a number of solvents have been conducted over a range of temperatures and pressures. The stability order for the five coordinate complex in the solvents investigated is CH₃CN>DMF>DMSO>C₆H₅CN> H₂O>ClCH₂CN at 25 °C. Differences in stability are considered in terms of the measured thermodynamic parameters ΔH° and ΔS°. Both steric and electronic factors were found to influence solvent coordination with the macrocyclic complex.For the equilibrium in CH₃CN, C₆H₅CN, DMF and H₂O, reaction volumes, ΔV°, of −3.2±0.5, −4.2±0.5, −0.2±0.5 and −0.5±0.5 cm³ mol⁻¹ respectively have been determined. Each is significantly smaller than the corresponding solvent molar volume. The ΔV° for the equilibrium in CH₃CN is comparable with the previously determined activation volume for exchange of this solvent on R, S, R, S- [Ni(tmc)(CH₃CN)]²⁺. The equilibrium and measured volume parameters are discussed in relation to the mechanism for solvent exchange.     

Macronutrient-specific effect of the MTNR1B genotype on lipid levels in response to 2 year weight-loss diets

Compelling evidence indicates that lipid metabolism is in partial control of the circadian system. In this context, it has been reported that the melatonin receptor 1B (MTNR1B) genetic variant influences the dynamics of melatonin secretion, which is involved in the circadian system as a chronobiotic. The objective was to analyze whether the MTNR1B rs10830963 genetic variant was related to changes in lipid levels in response to dietary interventions with different macronutrient distribution in 722 overweight/obese subjects from the POUNDS Lost trial. We did not find a significant association between the MTNR1B genotype and changes in lipid metabolism. However, dietary fat intake significantly modified genetic effects on 2 year changes in total and LDL cholesterol (P interaction = 0.006 and 0.001, respectively). In the low-fat diet group, carriers of the sleep disruption G allele (minor allele) showed a greater reduction of total cholesterol (β ± SE = −5.78 ± 2.88 mg/dl, P = 0.04) and LDL cholesterol (β ± SE = −7.19 ± 2.37 mg/dl, P = 0.003). Conversely, in the high-fat diet group, subjects carrying the G allele evidenced a smaller decrease in total cholesterol (β ± SE = 5.81 ± 2.65 mg/dl, P = 0.03) and LDL cholesterol (β ± SE = 5.23 ± 2.21 mg/dl, P = 0.002). Subjects carrying the G allele of the circadian rhythm-related MTNR1B variant may present a bigger impact on total and LDL cholesterol when undertaking an energy-restricted low-fat diet.     

A putative <Emphasis Type="Italic">NEM1</Emphasis> homologue regulates lipid droplet biogenesis <Emphasis Type="Italic">via PAH1</Emphasis> in <Emphasis Type="Italic">Tetrahymena thermophila</Emphasis>

Nuclear envelope morphology protein 1 (NEM1) along with a phosphatidate phosphatase (PAH1) regulates lipid homeostasis and membrane biogenesis in yeast and mammals. We investigated four putative NEM1 homologues (TtNEM1A, TtNEM1B, TtNEM1C and TtNEM1D) in the Tetrahymena thermophila genome. Disruption of TtNEM1B, TtNEM1C or TtNEM1D did not compromise normal cell growth. In contrast, we were unable to generate knockout strain of TtNEM1A under the same conditions, indicating that TtNEM1A is essential for Tetrahymena growth. Interestingly, loss of TtNEM1B but not TtNEM1C or TtNEM1D caused a reduction in lipid droplet number. Similar to yeast and mammals, TtNem1B of Tetrahymena exerts its function via Pah1, since we found that PAH1 overexpression rescued loss of Nem1 function. However, unlike NEM1 in other organisms, TtNEM1B does not regulate ER/nuclear morphology. Similarly, neither TtNEM1C nor TtNEM1D is required to maintain normal ER morphology. While Tetrahymena PAH1 was shown to functionally replace yeast PAH1 earlier, we observed that Tetrahymena NEM1 homologues did not functionally replace yeast NEM1. Overall, our results suggest the presence of a conserved cascade for regulation of lipid homeostasis and membrane biogenesis in Tetrahymena. Our results also suggest a Nem1-independent function of Pah1 in the regulation of ER morphology in Tetrahymena.