Conserved Residues R420 and Q428 in a Cytoplasmic Loop of the Citrate/Malate Transporter CimH of Bacillus subtilis Are Accessible from the External Face of the Membrane

CimH of Bacillus subtilis is a secondary transporter for citrate and malate that belongs to the 2-hydroxycarboxylate transporter (2HCT) family. Conserved residues R143, R420, and Q428, located in putative cytoplasmic loops and R432, located at the cytoplasmic end of the C-terminal transmembrane segment XI were mutated to Cys to identify residues involved in binding of the substrates. R143C, R420C, and Q428C revealed kinetics similar to those of the wild-type transporter, while the activity of R432C was reduced by at least 2 orders of magnitude. Conservative replacement of R432 with Lys reduced the activity by 1 order of magnitude, by lowering the affinity for the substrate 10-fold. It is concluded that the arginine residue at position 432 in CimH interacts with one of the carboxylate groups of the substrates. Labeling of the R420C and Q428C mutants with thiol reagents inhibited citrate transport activity. Surprisingly, the cysteine residues in the cytoplasmic loops in both R420C and Q428C were accessible to the small, membrane-impermeable, negatively charged MTSES reagent from the external site of the membrane in a substrate protectable manner. The membrane impermeable reagents MTSET, which is positively charged, and AMdiS, which is negatively charged like MTSES but more bulky, did not inhibit R420C and Q428C. It is suggested that the access pathway is optimized for small, negatively charged substrates. Either the cytoplasmic loop containing residues R420 and Q428 is partly protruding to the outside, possibly in a reentrant loop like structure, or alternatively, a water-filled substrate translocation pathway extents to the cytoplasm-membrane interface.

Continuing education: The 1998 survey of the Royal Australasian College of Dental Surgeons

The document attached has been archived with permission from the Australian Dental Association. An external link to the publisher’s copy is included.Background: Continuing education (CE) is an essential professional activity. In the last decade, CE has been actively pursued by the medical profession in Australia and abroad. However, the uptake of CE in dentistry has been much slower and there is minimal Australian data on dental CE. Methods: To determine the level of CE activity, in 1998, postal questionnaires were sent to all fellows of the Royal Australasian College of Dental Surgeons. The responses were analysed. Results: There was a high reponse rate (90 per cent) but a moderate usable rate (54 per cent). The results show a biphasic distribution between high and low CE activity. The average amount of activity of those involved in CE was 116 hours per year, above the usually accepted minimum of 100 hours/year. Some groups, particularly members of the specialist divisions of oral and maxillofacial surgeons (215 hours) and periodontists (205 hours), have high levels of CE. However, approximately 25 per cent of college fellows reported little or no CE activity. The survey revealed that inactive fellows are more likely to be older and in general practice. Inactive fellows were also tardy in replying to the questionnaire. Conclusion: The high activity CE group needs to be recognised and encouraged to continue. Specific plans to help the low CE activity group should be developed. Although these findings relate directly to the Royal Australasian College of Dental Surgeons, they are presented as they have implications for the dental profession at large.P Sambrook, D Thomson, R Bastiaan and A Gos

Too hot to handle: heat stroke in multiple system atrophy

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Exact Diffusion Inversion via Bi-directional Integration Approximation

Recently, different methods have been proposed to address the inconsistency issue of DDIM inversion to enable image editing, such as EDICT \cite{Wallace23EDICT} and Null-text inversion \cite{Mokady23NullTestInv}. However, the above methods introduce considerable computational overhead. In this paper, we propose a new technique, named \emph{bi-directional integration approximation} (BDIA), to perform exact diffusion inversion with neglible computational overhead. Suppose we would like to estimate the next diffusion state $\boldsymbol{z}_{i-1}$ at timestep $t_i$ with the historical information $(i,\boldsymbol{z}_i)$ and $(i+1,\boldsymbol{z}_{i+1})$. We first obtain the estimated Gaussian noise $\hat{\boldsymbol{\epsilon}}(\boldsymbol{z}_i,i)$, and then apply the DDIM update procedure twice for approximating the ODE integration over the next time-slot $[t_i, t_{i-1}]$ in the forward manner and the previous time-slot $[t_i, t_{t+1}]$ in the backward manner. The DDIM step for the previous time-slot is used to refine the integration approximation made earlier when computing $\boldsymbol{z}_i$. One nice property with BDIA-DDIM is that the update expression for $\boldsymbol{z}_{i-1}$ is a linear combination of $(\boldsymbol{z}_{i+1}, \boldsymbol{z}_i, \hat{\boldsymbol{\epsilon}}(\boldsymbol{z}_i,i))$. This allows for exact backward computation of $\boldsymbol{z}_{i+1}$ given $(\boldsymbol{z}_i, \boldsymbol{z}_{i-1})$, thus leading to exact diffusion inversion. Experiments on both image reconstruction and image editing were conducted, confirming our statement. BDIA can also be applied to improve the performance of other ODE solvers in addition to DDIM. In our work, it is found that applying BDIA to the EDM sampling procedure produces slightly better FID score over CIFAR10.Comment: arXiv admin note: text overlap with arXiv:2304.1132

Oligo pools as an affordable source of synthetic DNA for cost-effective library construction in protein- and metabolic pathway engineering

The construction of custom libraries is critical for rational protein engineering and directed evolution. Array-synthesized oligo pools of thousands of user-defined sequences (up to ~350 bases in length) have emerged as a low-cost commercially available source of DNA. These pools cost ≤10% (depending on error rate and length) of other commercial sources of custom DNA, and this significant cost difference can determine whether an enzyme engineering project can be realized on a given research budget. However, while being cheap, oligo pools do suffer from a low concentration of individual oligos and relatively high error rates. Several powerful techniques that specifically make use of oligo pools have been developed and proven valuable or even essential for next-generation protein and pathway engineering strategies, such as sequence-function mapping, enzyme minimization, or de-novo design. Here we consolidate the knowledge on these techniques and their applications to facilitate the use of oligo pools within the protein engineering community

A modal impedance-angle formalism: Rigorous proofs for optical fiber mode counting and bracketing

In a companion paper, a complex-power-flow variational scheme is applied to analyze mode propagation along open circularly cylindrical graded-index waveguides. It leads to a characteristic equation in terms of impedances rather than fields. The resulting impedance-angle formalism provides the basis for the full-wave generalization for optical fibers of the mode-counting scheme previously developed for a scalar wave propagation problem. The complex-power-flow variational scheme for bent waveguides is based on energy considerations. Hence, in its derivation, it is natural to consider a waveguide section (a volume) rather than a cross section (a surface). In the proof of the mode-counting and mode-bracketing theorems, the key issue is to show that the characteristic roots and the roots of the so-called separation function alternate. For general circularly cylindrical open waveguides, the required proofs are intricate. However, the special limiting cases in which the optical fiber is surrounded by electrically or magnetically perfectly conducting walls are tractable. To account for the general case, it appears to be necessary to regard a class of optical waveguide problems with a continuous transition from perfect electric conductor to perfect magnetic conductor boundary conditions via the situation pertaining to the actual exterior medium. Thus, a half-strip is constructed on which the so-called characteristic and separation graphs are seen to alternate. As spin-off, such a "sweep" might prove useful in the design of a fiber cladding

The clinical heterogeneity of drug-induced myoclonus: an illustrated review

Contains fulltext : 177995.pdf (publisher's version ) (Open Access)A wide variety of drugs can cause myoclonus. To illustrate this, we first discuss two personally observed cases, one presenting with generalized, but facial-predominant, myoclonus that was induced by amantadine; and the other presenting with propriospinal myoclonus triggered by an antibiotic. We then review the literature on drugs that may cause myoclonus, extracting the corresponding clinical phenotype and suggested underlying pathophysiology. The most frequently reported classes of drugs causing myoclonus include opiates, antidepressants, antipsychotics, and antibiotics. The distribution of myoclonus ranges from focal to generalized, even amongst patients using the same drug, which suggests various neuro-anatomical generators. Possible underlying pathophysiological alterations involve serotonin, dopamine, GABA, and glutamate-related processes at various levels of the neuraxis. The high number of cases of drug-induced myoclonus, together with their reported heterogeneous clinical characteristics, underscores the importance of considering drugs as a possible cause of myoclonus, regardless of its clinical characteristics

Transporters involved in uptake of di- and tricarboxylates in Bacillus subtilis

Di- and tricarboxylates found as intermediates in the tricarboxylic acid cycle can be utilized by many bacteria and serve as carbon and energy source under aerobic and anaerobic conditions. A prerequisite for metabolism is that the carboxylates are transported into the cells across the cytoplasmic membrane. Bacillus subtilis is able to metabolize many di- and tricarboxylates and in this overview the available data on all known and putative di- and tricarboxylate transporters in B. subtilis is summarized. The B. subtilis transporters, that are of the secondary type, are discussed in the context of the protein families to which they belong. Available data on biochemical characterization, regulation of gene expression and the physiological function is summarized. It is concluded that in B. subtilis multiple transporters are present for tricarboxylic acid cycle intermediates.</p