Detecting envelope stress by monitoring ß-barrel assembly.

The cell envelope protects bacteria from their surroundings. Defects in its integrity or assembly are sensed by signal transduction systems, allowing cells to rapidly adjust. The Rcs phosphorelay responds to outer membrane (OM)- and peptidoglycan-related stress in enterobacteria. We elucidated how the OM lipoprotein RcsF, the upstream Rcs component, senses envelope stress and activates the signaling cascade. RcsF interacts with BamA, the major component of the ß-barrel assembly machinery. In growing cells, BamA continuously funnels RcsF through the ß-barrel OmpA, displaying RcsF on the cell surface. This process spatially separates RcsF from the downstream Rcs component, which we show is the inner membrane protein IgaA. The Rcs system is activated when BamA fails to bind RcsF and funnel it to OmpA. Newly synthesized RcsF then remains periplasmic, interacting with IgaA to activate the cascade. Thus RcsF senses envelope damage by monitoring the activity of the Bam machinery.

Disorder is a critical component of lipoprotein sorting in Gram-negative bacteria.

Gram-negative bacteria express structurally diverse lipoproteins in their cell envelope. Here, we find that approximately half of lipoproteins destined to the Escherichia coli outer membrane display an intrinsically disordered linker at their N terminus. Intrinsically disordered regions are common in proteins, but establishing their importance in vivo has remained challenging. As we sought to unravel how lipoproteins mature, we discovered that unstructured linkers are required for optimal trafficking by the Lol lipoprotein sorting system, whereby linker deletion re-routes three unrelated lipoproteins to the inner membrane. Focusing on the stress sensor RcsF, we found that replacing the linker with an artificial peptide restored normal outer-membrane targeting only when the peptide was of similar length and disordered. Overall, this study reveals the role played by intrinsic disorder in lipoprotein sorting, providing mechanistic insight into the biogenesis of these proteins and suggesting that evolution can select for intrinsic disorder that supports protein function.

Repairing oxidized proteins in the bacterial envelope using respiratory chain electrons.

The reactive species of oxygen and chlorine damage cellular components, potentially leading to cell death. In proteins, the sulfur-containing amino acid methionine is converted to methionine sulfoxide, which can cause a loss of biological activity. To rescue proteins with methionine sulfoxide residues, living cells express methionine sulfoxide reductases (Msrs) in most subcellular compartments, including the cytosol, mitochondria and chloroplasts. Here we report the identification of an enzymatic system, MsrPQ, repairing proteins containing methionine sulfoxide in the bacterial cell envelope, a compartment particularly exposed to the reactive species of oxygen and chlorine generated by the host defence mechanisms. MsrP, a molybdo-enzyme, and MsrQ, a haem-binding membrane protein, are widely conserved throughout Gram-negative bacteria, including major human pathogens. MsrPQ synthesis is induced by hypochlorous acid, a powerful antimicrobial released by neutrophils. Consistently, MsrPQ is essential for the maintenance of envelope integrity under bleach stress, rescuing a wide series of structurally unrelated periplasmic proteins from methionine oxidation, including the primary periplasmic chaperone SurA. For this activity, MsrPQ uses electrons from the respiratory chain, which represents a novel mechanism to import reducing equivalents into the bacterial cell envelope. A remarkable feature of MsrPQ is its capacity to reduce both rectus (R-) and sinister (S-) diastereoisomers of methionine sulfoxide, making this oxidoreductase complex functionally different from previously identified Msrs. The discovery that a large class of bacteria contain a single, non-stereospecific enzymatic complex fully protecting methionine residues from oxidation should prompt a search for similar systems in eukaryotic subcellular oxidizing compartments, including the endoplasmic reticulum.

Modified headspace solid-phase microextraction for the determination of quantitative relationships between components of mixtures consisting of alcohols, esters, and ethers - impact of the vapor pressure difference of the compounds.

The quantitative relationship between analytes established by the headspace solid-phase microextraction procedure for multicomponent mixtures depends not only on the character and strength of interactions of individual components with solid-phase microextraction fiber but also on their vapor pressure in the applied headspace solid-phase microextraction system. This study proves that vapor pressure is of minor importance when the sample is dissolved/suspended in a low-volatility liquid of the same physicochemical character as that of the used solid phase microextraction fiber coating. It is demonstrated for mixtures of alcohols, esters, ethers and their selected representatives by applying a headspace solid-phase microextraction system composed of Carbowax fiber and sample solutions in polyethyleneglycol. The observed differences in quantitative relations between components of the examined mixtures established by their direct analysis and by modified headspace solid-phase microextraction are insignificant (Fexp  < Fcrit ). It is explained by a significant diminution in vapor pressure difference between individual components of the examined mixture in the applied headspace solid phase microextraction system due to low components concentration in polyethyleneglycol suspensions (Raoult's law) and due to strong specific interactions of analyte molecules with polyethyleneglycol molecules.

Unilateral optic disc oedema as a first sign of ocular toxoplasmosis ­ case report. / Jednostronny obrzek tarczy nerwu wzrokowego jako pierwszy objaw toksoplazmozy ocznej ­ analiza przypadku.

Diagnostic procedure for unilateral optic disc oedema needs detailed medical history, careful physical examination and multiple additional tests. We present a very rare case of atypical unilateral anterior optic neuropathy as a sign of ocular toxoplasmosis in a young female. All diagnostic and therapeutic challenges have been discussed.

[Micropulse 577 nm laser trabeculoplasty--a prospective pilot study]. / Trabekuloplastyka laserowa w trybie mikropulsów laserem 577 nm--pilotazowe badanie prospektywne.

PURPOSE: To evaluate the efficacy of micropulse 577 nm laser trabeculoplasty. MATERIAL AND METHODS: Prospective clinical trial designed for 30 patients diagnosed with open angle glaucoma. The micropulse 577 nm laser trabeculoplasty was performed in one eye of each patient in the lower half of trabecular meshwork. The intraocular pressure was measured using two independent methods--Pascal tonometer and iCare tonometer--at baseline (before surgery), at 3 h, 24 h, 2 weeks and 1 month following the procedure. Pressure measurement results were compared between the treated and untreated eyes. Treatment was considered successful if the intraocular pressure dropped by ≥ 3 mmHg or ≥ 20% as compared to the baseline values or ≥ 20% as compared to the fellow eye. RESULTS: The study was conducted on 11 patients. In the treated group, Pascal tonometry showed the mean decrease of the intraocular pressure by 1.7 mmHg (7.5%). The same value for the untreated eyes was 1.8 mmHg (8.1%). The results obtained using iCare tonometer showed the same proportions. The treatment did not lower intraocular pressure as compared to the untreated fellow eyes. There were no adverse effects. Even though it was impossible to perform the proper statistical analysis due to the small number of treated patients the study was eventually interrupted for ethical reasons, because of the failure to decrease the intraocular pressure in the treated group of 11 patients. CONCLUSIONS: The treatment appeared to be safe but ineffective in the treated group. Further research is needed to modify laser parameters for the improvedefficacy and the maintained good safety profile of the procedure.

The Cryptosporidium signaling kinase CDPK5 plays an important role in male gametogenesis and parasite virulence.

The protozoan parasite Cryptosporidium is a leading cause of diarrhea in young children. The parasite's life cycle involves a coordinated and timely progression from asexual to sexual stages, leading to the formation of the transmissible oocyst. Underlying molecular signaling mechanisms orchestrating sexual development are not known. Here, we describe the function of a signaling kinase in Cryptosporidium male gametogenesis. We reveal the expression of Cryptosporidium parvum calcium-dependent protein kinase 5 (CDPK5) during male gamete development and its important role in the egress of mature gametes. Genetic ablation of this kinase results in viable parasites, indicating that this gene is dispensable for parasite survival. Interestingly, cdpk5 deletion decreases parasite virulence and impacts oocyst shedding in immunocompromised mice. Using phosphoproteomics, we identify possible CDPK5 substrates and biological processes regulated by this kinase. Collectively, these findings illuminate parasite cell biology by revealing a mechanism controlling male gamete production and a potential target to block disease transmission.

Overdosage Section in US and EU Labeling.

The Prescribing Information (PI) in the United States (US) and the Summary of Product Characteristics (SmPC) in the European Union (EU) are approved by the US Food & Drug Administration (FDA), and the European Medicines Agency (EMA), respectively. The inclusion of overdosage information in these documents is a regulatory requirement in both regions. This research evaluates the content of the overdosage section of US and EU labeling. The overdosage sections of labels for drugs approved in the US in three time periods were analyzed: 2000-2001, 2010-2011, and 2020-2021. EU labels for these same products were also reviewed if registered through the Centralized Procedure. Data collection and analyses were performed using a predefined questionnaire, focusing on adherence to regulatory requirements and identifying areas where additional regulatory guidance may be beneficial. The findings indicate that the content of the overdosage sections largely comply with the regulatory requirements of their respective regions. Fewer than half of the labels included information on supratherapeutic doses observed from clinical studies, risk factors for overdose or population specific data associated with overdose. Inconsistencies were noted concerning the incorporation of animal data when human data were available, in addition to the referencing of Poison Centers. The overall utility of non-specific treatment recommendations, in addition to gastric lavage is discussed. While the content of the overdosage section generally aligns with regulatory expectations, additional regulatory guidance could enhance consistency in how this section of labeling is presented and clarify expectations to improve its usefulness for health care professionals (HCPs).

The Feasibility of Surface Electromyography in Monitoring Orbicularis Oculi Recovery after Anterior Approach Levator Aponeurosis Advancement.

INTRODUCTION: In this article, we propose a new application for eyelid surface electromyography (sEMG). By placing the electrode in the mid-pretarsal area of the upper eyelid, one can easily perform a fast examination and achieve repeatable results. We believe that this technique may increase the feasibility of eyelid sEMG in clinical practice. METHODS: 126 sEMG examinations of the upper eyelid were performed by using the above-described method. Thirty-nine controls and 29 ptotic patients were enrolled. The controls underwent one measurement while the ptotic patients were employed for four sessions: Before anterior approach levator aponeurosis advancement (LAA), 2 weeks, 3 months, and more than 6 months after surgery. The relaxation and maximal contraction of the orbicularis oculi muscle (OOM) using root mean square (RMS) values were measured. RESULTS: The results showed a statistically significant decrease in RMS values of the maximal contraction of the OOM 2 weeks after surgery (p < 0.05) and 3 months after surgery (p = 0.03). Six months postoperatively, there were no statistically significant differences in OOM activity compared to preoperative values (p = 0.2). CONCLUSIONS: Eyelid sEMG may be a useful diagnostic tool in post-operative OOM recovery monitoring. sEMG parameters of the maximal contraction of the OOM normalize within 6 months after anterior approach LAA. Electrode placement in the mid-pretarsal area of the upper eyelid offers several advantages and therefore may enhance the feasibility of sEMG in clinical practice.

Itraorbital organic foreign body - radiological methods in diagnosis - case report.

Intraorbital foreign bodies, especially non-metallic ones, remain an important diagnostic and therapeutic problem. Organic foreign bodies contain large quantities of bacterial flora contributing to rapid onset of inflammation which may promptly damage the optic nerve. Further progression of inflammation may involve the structures of central nervous system. US examination is a preliminary, commonly available diagnostic method, although it is not always successful in visualizing foreign bodies. CT scan remains a standard investigation used to visualize intraorbital foreign bodies, although with organic foreign bodies, its results may be inconclusive. MRI allows for visualization and precise identification of foreign bodies, which is vital for surgery. The issue of intraorbital foreign bodies is an interdisciplinary problem requiring the co-operation between ophthalmologists and neurosurgeons.

[Nursing care in perioperative period in patients with intenstinal stomia exposure]. / Opieka pielegniarska w okresie okolooperacyjnym nad pacjentem z wyloniona stomia jelitowa.

Despite of enormous advance in minimally invasive surgery which is almost scarless nowadays, there is still very important emotional issue for patients connected with each surgical procedure. One of the most stressful surgical procedures for patients is the one which ends up with stomia exposure. The main objective of this article is to point out very the important factor which leads to decrease the number of complications, speeds up recovery and acceptation of the stomia by patients. This factor is known as a professional nursing care. It consists of physical and psychical preoperative preparation and postoperative care for patients. Special care in early postoperative 24h is crucial for preventing from development of any complications. That is why the nursing personnel is obliged to monitor vital signs very carefully. Complex preparation and postoperative care leads to diminish significantly the number of complications, facilitates cooperation with patients and also influences the increase of sense of safety and trust to medical personnel. Patients with stomia who were under professional nursing staff supervision achieve full recovery and higher quality of life considerably earlier.

Modified application of HS-SPME for quality evaluation of essential oil plant materials.

The main limitation in the standard application of head space analysis employing solid phase microextraction (HS-SPME) for the evaluation of plants as sources of essential oils (EOs) are different quantitative relations of EO components from those obtained by direct analysis of EO which was got in the steam distillation (SD) process from the same plant (EO/SD). The results presented in the paper for thyme, mint, sage, basil, savory, and marjoram prove that the quantitative relations of EO components established by HS-SPME procedure and direct analysis of EO/SD are similar when the plant material in the HS-SPME process is replaced by its suspension in oil of the same physicochemical character as that of SPME fiber coating. The observed differences in the thyme EO composition estimated by both procedures are insignificant (F(exp)

Modified HS-SPME for determination of quantitative relations between low-molecular oxygen compounds in various matrices.

Similar quantitative relations between individual constituents of the liquid sample established by its direct injection can be obtained applying Polydimethylsiloxane (PDMS) fiber in the headspace solid phase microextraction (HS-SPME) system containing the examined sample suspended in methyl silica oil. This paper proves that the analogous system composed of sample suspension/emulsion in polyethylene glycol (PEG) and Carbowax fiber allows to get similar quantitative relations between components of the mixture as those established by its direct analysis, but only for polar constituents. It is demonstrated for essential oil (EO) components of savory, sage, mint and thyme, and of artificial liquid mixture of polar constituents. The observed differences in quantitative relations between polar constituents estimated by both applied procedures are insignificant (Fexp < Fcrit). The presented results indicates that wider applicability of the system composed of a sample suspended in the oil of the same physicochemical character as that of used SPME fiber coating strongly depends on the character of interactions between analytes-suspending liquid and analytes-fiber coating.

Asymmetric Synthesis and Properties of Sulfinimines (Thiooxime S-Oxides).

Enantiomerically pure sulfinimines (thiooxime S-oxides 10), important building blocks in the asymmetric synthesis of amine derivatives, are prepared in good to excellent yields in one step from aromatic, heteroaromatic, and aliphatic aldehydes. This protocol involves treating commercially available (R)- or (S)-menthyl p-toluenesufinate (Andersen reagent 4) with LiHMDS, followed by the aldehyde, affording (E)-10 exclusively. The sulfinimines 10 are formed via a Peterson-type olefination reaction of silylsulfinamide anion 13 with the aldehyde. Anion 13 is generated by reaction of lithium menthoxide (12a) with bis(trimethylsilyl)sulfinamide 11, which is formed in the reaction of 4 with LiHMDS. The other product formed is O-(trimethylsilyl)menthol (12c), which is isolated in >80% yield for recycling. Two other less efficient methods for the asymmetric synthesis of 10 are discussed: (i) the asymmetric oxidation of sulfenimines 6 with chiral nonracemic oxaziridines and (ii) the reaction of metal aldimines, prepared from nitriles, with 4. All of these protocols fail with ketones.

Acetylsalicylic acid (ASA) use among Polish adults with diabetes mellitus.

UNLABELLED: People with diabetes mellitus have higher risk of cardiovascular morbidity and mortality from thrombo-vascular complications than non-diabetics and it is recommended that they should use acetylsalicylic acid (ASA) as anitiplatelet agent regularly. The aim of this study was to examine current physician counseling about ASA therapy and the use of ASA by Polish patients with diabetes. The study involved 464 randomly selected Caucasian diabetic individuals, mainly with type 2 (>98%), who were asked to complete a questionnaire consisting of 16 questions while visiting out-patient medical centres for periodical examination or during hospitalization. We collected data on the prevalence of cardiovascular disease (CVD), frequency of risk factors among diabetic patients and on ASA physician counseling, and on the rate of regular ASA users among the patients. The most self-reported conditions were angina pectoris or prior myocardial infarction (63,6%). Current cigarette smoking was declared by 7,1% of the patients, elevated serum cholesterol levels by 58%, and hypertension by 62,9%. Majority of the patients were overweight, and family history of coronary artery disease (CAD) was reported by 30,2% of the patients. ASA was taken by 67,2% of all the patients and 72,1% of them had taken ASA regularly. The highest percentage (>80%) of regular ASA users was found among diabetic patients with CAD. A minority of the patients who reported stroke (43,9%) or claudication (15,8%) were taking ASA regularly. Number of the patients (32,8%) with CVD risk factor(s) but without diagnosed CVD did not use ASA at all. The majority of the patients (78%) were counseled by their physicians why they should use ASA regularly. IN CONCLUSION: the number of regular users of ASA among diabetic patients in Poland with CVD others than CAD or among patients with at least two CAD risk factors is still unsatisfactory. Therefore, while it seems that there is a remarkable growing of ASA therapy world-wide more effort is needed to make this practice a standard of multifactorial, intensive diabetes treatment. Our results also suggest the need for more aggressive education of both doctors and patients.