Outcomes of 11 dogs with short radius syndrome treated with acute arthroscopically assisted ulnar shortening.

OBJECTIVE: To describe the short-term outcome of acute arthroscopically assisted ulnar shortening (AUS), to treat short radius syndrome in dogs. STUDY DESIGN: Case series. ANIMALS: Eleven client owned dogs. METHODS: Records of dogs that had undergone AUS for treatment of short radius syndrome were reviewed for inclusion. Reporting data included among others pre- and postoperative radioulnar, humeroradial and humeroulnar distances, lameness scores, surgical times, complications and clinical outcome. RESULTS: Following AUS, radiohumeral articulation was improved in all dogs. Median presurgery radioulnar, humeroradial and humeroulnar values were 4.5, 3.2, and 2.2 mm and were improved with surgery by a median of 3.2, 1.8, and 1.2 mm, respectively. Median surgery time was 140 min. Median time to bone healing was 8 weeks (range: 4-14). Median time to last follow-up was 9 weeks (4-468). Median lameness score (scale 0-4) improved from 2 to 1. No major complications were reported. Short-term clinical outcome was graded by the surgeons as full function in four cases and acceptable function in seven. CONCLUSION AND CLINICAL RELEVANCE: Radiographic and arthroscopic radiohumeral articulation were improved and short-term clinical improvement was documented following AUS in all 11 dogs.

Is pitolisant safe for clinical use? A retrospective pharmacovigilance study focus on the post-marketing safety.

Pitolisant, a novel histamine H3-receptor antagonist, holds significant promise for treating narcolepsy. However, a petition, which highlighted that pitolisant was associated with deaths during clinical trials, has propelled it into the spotlight of widespread societal attention on April 3, 2023. Till now, the clinical safety of pitolisant remains a heatedly debated topic. This study aimed to offer a comprehensive assessment of the safety profile of pitolisant in real-world clinical settings. Adverse event reports where pitolisant was the primary suspect drug were extracted from the FDA Adverse Event Reporting System database. The clinical characteristics and concomitant drugs of the pitolisant-associated adverse events were analyzed. The potential adverse event signals of pitolisant were explored using four disproportionality analysis methods. Furthermore, the difference in pitolisant-associated adverse event signals was investigated concerning sex, age, weight, and dose. A total of 526 reports and 1695 adverse events with pitolisant as the primary suspected drug were identified. The most significant adverse event signals were generally mild and of short duration. The concomitant drugs of pitolisant were highly intricate, mainly included drugs for treating narcolepsy as well as antidepressants. Seven new significant adverse event signals emerged. The safety profile of pitolisant exhibited no significant differences across age and dose groups, although slight variations were observed in relation to sex and weight. The findings from reports of death and life-threatening outcomes underscore the importance of enhanced monitoring for cardiac and respiratory adverse reactions when utilizing pitolisant. This study provided a broader understanding of the safety profile of pitolisant.

Pre-optimization and one-step preparation of cascade enzymes system with broad substrates by model guidance: Application of chiral L-norvaline and L-phenylglycine biosynthesis.

Cascade biocatalyst systems with catalytic promiscuity can be used for synthesis of a class of chiral chemicals but the optimization of these systems by model guidance is poorly explored. In this study, a cascade system with broad substrate spectrum was characterized and simulated by kinetic model with substrates of DL-Norvaline (DL-Nor) and DL-Phenylglycine (DL-Phg) as examples. To evaluate the optimal cascade system, maximum accumulation of intermediate products and conversion rate in the process were investigated by simultaneous solution of the rate equations for varying enzyme quantities. According to the simulation results, the cascade system was optimized by regulating the expression of D-amino acid oxidase and formate dehydrogenase and was prepared by one-step. The conversion efficiency of DL-Nor and DL-Phg have been significantly improved compared with that of before optimization. Moreover, the total of L-Nor and L-Phg were reached 498.2 mM and 79.5 mM through a gradient fed-batch conversion strategy, respectively.

Adrenal tumors treated by adrenalectomy following spontaneous rupture carry an overall favorable prognosis: retrospective evaluation of outcomes in 59 dogs and 3 cats (2000-2021).

OBJECTIVE: To conduct a retrospective multi-institutional study reporting short- and long-term outcomes of adrenalectomy in patients presenting with acute hemorrhage secondary to spontaneous adrenal rupture. ANIMALS: 59 dogs and 3 cats. METHODS: Medical records of dogs and cats undergoing adrenalectomy between 2000 and 2021 for ruptured adrenal masses were reviewed. Data collected included clinical presentation, preoperative diagnostics, surgical report, anesthesia and hospitalization findings, histopathology, adjuvant treatments, and long-term outcome (recurrence, metastasis, and survival). RESULTS: Median time from hospital admission to surgery was 3 days, with 34% of surgeries being performed emergently (within 1 day of presentation). Need for intraoperative blood transfusion was significantly associated with emergent surgery and presence of active intraoperative hemorrhage. The short-term (≤ 14 days) complication and mortality rates were 42% and 21%, respectively. Negative prognostic factors for short-term survival included emergent surgery, intraoperative hypotension, and performing additional surgical procedures. Diagnoses included adrenocortical neoplasia (malignant [41%], benign [12%], and undetermined [5%]), pheochromocytoma (38%), a single case of adrenal fibrosis and hemorrhage (2%), and a single case of hemangiosarcoma (2%). Local recurrence and metastasis of adrenocortical carcinoma were confirmed in 1 and 3 cases, respectively. Overall median survival time was 574 days and 900 days when short-term mortality was censored. No significant relationship was found between histopathological diagnosis and survival. CLINICAL RELEVANCE: Adrenalectomy for ruptured adrenal gland masses was associated with similar short- and long-term outcomes as compared with previously reported nonruptured cases. If hemodynamic stability can be achieved, delaying surgery and limiting additional procedures appear indicated to optimize short-term survival.

Incidence, survival time, and surgical treatment of parathyroid carcinomas in dogs: 100 cases (2010-2019).

OBJECTIVE: To evaluate outcomes of dogs with parathyroid carcinoma (PTC) treated by surgical excision and to describe the incidence of postoperative hypocalcemia, degree of hypocalcemia, duration of hospitalization, duration of calcium supplementation, and survival time. ANIMALS: 100 client-owned dogs with PTC admitted to academic, referral veterinary institutions. PROCEDURES: In a retrospective multi-institutional study, medical records of dogs undergoing surgical excision of PTC between 2010 to 2019 were reviewed. Signalment, relevant medical history, clinical signs, clinicopathologic testing, imaging, surgical findings, intraoperative complications, histologic examination, and survival time were recorded. RESULTS: 100 dogs with PTC were included, and 96 dogs had clinical or incidental hypercalcemia. Common clinical signs included polyuria (44%), polydipsia (43%), hind limb paresis (22%), lethargy (21%), and hyporexia (20%). Cervical ultrasonography detected a parathyroid nodule in 91 of 91 dogs, with a single nodule in 70.3% (64/91), 2 nodules in 25.3% (23/91), and ≥ 3 nodules in 4 (4/91)% of dogs. Hypercalcemia resolved in 89 of 96 dogs within 7 days after surgery. Thirty-four percent of dogs developed hypocalcemia, on the basis of individual analyzer ranges, within 1 week after surgery. One dog had metastatic PTC to the prescapular lymph node, and 3 dogs were euthanized for refractory postoperative hypocalcemia. Estimated 1-, 2-, and 3-year survival rates were 84%, 65%, and 51% respectively, with a median survival time of 2 years. CONCLUSIONS AND CLINICAL RELEVANCE: Excision of PTC results in resolution of hypercalcemia and excellent long-term tumor control. Surgical excision of PTC is recommended because of resolution of hypercalcemia and a good long-term prognosis. Future prospective studies and long-term follow-up are needed to further assess primary tumor recurrence, metastasis, and incidence of postoperative hypocalcemia.

Characterization of Dendrolimus houi Lajonquiere (Lepidoptera: Lasiocampidae) Transcriptome across All Life Stages.

Dendrolimus houi Lajonquiere is a phytophagous caterpillar infesting many economically important coniferous tree species in China, causing serious economic and ecological environment losses. Based on previous research, it has one generation per year in South China and East China in contrast to two generations per year in Yunnan province in southwestern China. The species is potentially resilient to climatic extremes in these regions with the eggs and 1st instar larvae surviving in the winter (5 °C), older instar larvae and pupae surviving high temperatures in the summer (35 °C), suggesting some temperature stress tolerance during different developmental stages. However, little is known in this species at the genetic and genomic level. In this study, we used high throughput sequencing to obtain transcriptome data from different developmental stages (eggs, 1st-3rd instar larvae, 4th-5th instar larvae, 6th-7th instar larvae, pupae, male and female adults), which were collected from Fujian province. In total, we obtained approximately 90 Gb of data, from which 33,720 unigenes were assembled and 17,797 unigenes were annotated. We furtherly analyzed the differentially expressed genes (DGEs) across all stages, the largest number between the eggs and 1st instar larvae stage and gene expression varied significantly in different developmental stages. Furthermore, 4138 SSR genes and 114,977 SNP loci were screened from transcriptome data. This paper will be a foundation for further study towards improved integrated pest management strategies for this species.

Rational Engineering of Bacillus cereus Leucine Dehydrogenase Towards α-keto Acid Reduction for Improving Unnatural Amino Acid Production.

Unnatural amino acids (UAAs) play a key role in modern medicinal chemistry such as small molecules and peptide-based drugs with fast-growing markets. Low efficiency for natural enzymes including leucine dehydrogenase (LeuDH, EC1.4.1.9) are one major challenge for UAA production. Here, rational engineering of LeuDH from Bacillus cereus with a structure-based design approach is studied. The results achieve higher enzymatic activity and stability toward α-keto acid reduction by improving the hydrophobic and rigidity of enzymatic substrate entrance tunnel. High catalytic efficiency for variant E116V is associated with the presence of more hydrophobic tunnels that allows easy substrate diffusion, which is confirmed in absorbance spectroscopy study. For variant T45M/E116V, melting temperature and half-lives of thermal inactivation at 60 °C is 62.8 °C and 29.2 h, respectively, much higher than 48.4 °C and 3.4 h of wild type. Structural analysis indicates that an additional hydrogen bond in ß5 fold is formed in variant T45M, which results in a more rigid ß5 fold leading to better stability. Furthermore, asymmetric synthesis of α-amino acids with coenzyme regeneration reveals higher productivities for variant T45M/E116V. This study indicates the importance of substrate entrance tunnel for enzymatic activities and stability, the engineered LeuDH would better serve UAA production.

[Whole-cell biotransformation for simultaneous synthesis of L-2-aminobutyric acid and D-gluconic acid in recombinant Escherichia coli].

A whole-cell catalyst using Escherichia coli BL21(DE3) as a host, expressing L- threonine dehydratase from Escherichia coli, and co-expressing leucine dehydrogenase from Bacillus cereus and glucose dehydrogenase from Bacillus subtilis for cofactor regeneration, was constructed and used for one-pot production of L-2-aminobutyric acid (L-ABA) and D- gluconic acid from L-threonine and D-glucose. We used shake-flask culture to study the whole-cell catalytic condition including temperature, pH, proper permeabilization of cells and optimal wet cells amount. Moreover, the whole-cell catalyst was cultured in 5-L fermentor by fed-batch fermentation, and 164 g/L L-threonine and 248 g/L D-glucose were converted to 141.6 g/L L-ABA and 269.4 g/L D-gluconic acid. The whole-cell catalyst is promising to fulfill industrial requirements for L-ABA and D-gluconic acid.

Efficient production of d-amino acid oxidase in Escherichia coli by a trade-off between its expression and biomass using N-terminal modification.

Native d-amino acid oxidase (DAAO) that is expressed mostly as inclusion body and its toxicity for E. coli hamper efficient heterologous expression. In this study, the soluble expression of DAAO from Rhodosporidium toruloides (RtDAAO) was improved in E. coli through N-terminal modification, but the cell biomass was decreased. Then a trade-off between DAAO expression and biomass was achieved to obtain the highest volumetric activity of DAAO through regulated the number of N-terminus histidine residues. When variant 2H3G was fused with three N-terminus histidine residues, the volumetric activity was increased by 3.1 times and the biomass was not significant change compared with the wild type. Finally, the N-terminus disordered region of RtDAAO (HSQK) was replaced with HHHG and the variant enzyme activity reached 80.7U/mL (with a 40 percent of inactive DAAO reduced) in a 7.5L fermenter in 24h.

Local melatonin application induces cold tolerance in distant organs of Citrullus lanatus L. via long distance transport.

Melatonin is a ubiquitous chemical substance that regulates plant growth and responses to stress. Several recent studies show that exogenous melatonin confers cold tolerance to plants; however, the underlying mechanisms remain largely unknown. Here, we report that melatonin application at optimal dose, either on the leaves or the roots, not only induced cold stress tolerance in the site of application, but also systemically induced cold tolerance in untreated distant parts. Foliar or rhizospheric treatment with melatonin increased the melatonin levels in untreated roots or leaves, respectively, under both normal and cold stress conditions, whereas rhizospheric melatonin treatment increased the melatonin exudation rates from the xylem. An increased accumulation of melatonin accompanied with an induction in antioxidant enzyme activity in distant untreated tissues alleviated cold-induced oxidative stress. In addition, RNA-seq analysis revealed that an abundance of cold defense-related genes involved in signal sensing and transduction, transcriptional regulation, protection and detoxification, and hormone signaling might mediate melatonin-induced cold tolerance. Taken together, our results suggest that melatonin can induce cold tolerance via long distance signaling, and such induction is associated with an enhanced antioxidant capacity and optimized defense gene expression. Such a mechanism can be greatly exploited to benefit the agricultural production.

Elimination of a Free Cysteine by Creation of a Disulfide Bond Increases the Activity and Stability of Candida boidinii Formate Dehydrogenase.

NAD+-dependent formate dehydrogenase (FDH; EC 1.2.1.2) is an industrial enzyme widely used for NADH regeneration. However, enzyme inactivation caused by the oxidation of cysteine residues is a flaw of native FDH. In this study, we relieved the oxidation of the free cysteine of FDH from Candida boidinii (CboFDH) through the construction of disulfide bonds between A10 and C23 as well as I239 and C262. Variants A10C, I239C, and A10C/I239C were obtained by the site-directed mutagenesis and their properties were studied. Results showed that there were no significant changes in the optimum temperature and pH between variants and wild-type CboFDH. However, the stabilities of all variant enzymes were improved. Specifically, the CboFDH variant A10C (A10Cfdh) showed a significant increase in copper ion resistance and acid resistance, a 6.7-fold increase in half-life at 60°C, and a 1.4-fold increase in catalytic efficiency compared with the wild type. Asymmetric synthesis of l-tert-leucine indicated that the process time was reduced by 40% with variant A10Cfdh, which benefited from the increase in catalytic efficiency. Circular dichroism analysis and molecular dynamics simulation indicated that variants that contained disulfide bonds lowered the overall root mean square deviation (RMSD) and consequently increased the protein rigidity without affecting the secondary structure of enzyme. This work is expected to provide a viable strategy to avoid the microbial enzyme inactivation caused by the oxidation of the free cysteine residues and improving their performances. IMPORTANCE: FDH is widely used for NADH regeneration in dehydrogenase-based synthesis of optically active compounds to decrease the cost of production. This study highlighted a viable strategy that was used to eliminate the oxidation of free cysteine residues of FDH from Candida boidinii by the introduction of disulfide bonds. Using this strategy, we obtained a variant FDH with improved activity and stability. The improvement of activity and stability of FDH is expected to reduce its price and then further to decrease the cost of its application.

Regulation of Plant Growth, Photosynthesis, Antioxidation and Osmosis by an Arbuscular Mycorrhizal Fungus in Watermelon Seedlings under Well-Watered and Drought Conditions.

Drought stress has become an increasingly serious environmental issue that influences the growth and production of watermelon. Previous studies found that arbuscular mycorrhizal (AM) colonization improved the fruit yield and water use efficiency (WUE) of watermelon grown under water stress; however, the exact mechanisms remain unknown. In this study, the effects of Glomus versiforme symbiosis on the growth, physio-biochemical attributes, and stress-responsive gene expressions of watermelon seedlings grown under well-watered and drought conditions were investigated. The results showed that AM colonization did not significantly influence the shoot growth of watermelon seedlings under well-watered conditions but did promote root development irrespective of water treatment. Drought stress decreased the leaf relative water content and chlorophyll concentration, but to a lesser extent in the AM plants. Compared with the non-mycorrhizal seedlings, mycorrhizal plants had higher non-photochemical quenching values, which reduced the chloroplast ultrastructural damage in the mesophyll cells and thus maintained higher photosynthetic efficiency. Moreover, AM inoculation led to significant enhancements in the enzyme activities and gene expressions of superoxide dismutase, catalase, ascorbate peroxidase, glutathione reductase, and monodehydroascorbate reductase in watermelon leaves upon drought imposition. Consequently, AM plants exhibited lower accumulation of MDA, H2O2 and [Formula: see text] compared with non-mycorrhizal plants. Under drought stress, the soluble sugar and proline contents were significantly increased, and further enhancements were observed by pre-treating the drought-stressed plants with AM. Taken together, our findings indicate that mycorrhizal colonization enhances watermelon drought tolerance through a stronger root system, greater protection of photosynthetic apparatus, a more efficient antioxidant system and improved osmoregulation. This study contributes to advances in the knowledge of AM-induced drought tolerance.

Radix Tetrastigma hemsleyani flavone inhibits proliferation, migration, and invasion of human lung carcinoma A549 cells.

Radix Tetrastigma hemsleyani flavone (RTHF) is widely used as a traditional herb and has detoxification and anti-inflammatory effects. In this study, we investigated the potential effects of RTHF on the growth and metastasis of human lung adenocarcinoma A549 cells and evaluated its mechanisms. A549 cells were treated with RTHF at various concentrations for different periods. In vitro Cell Counting Kit-8 assay and colony formation methods showed that RTHF had dose- and time-dependent antiproliferation effects on A549 cells. A cell adhesion assay showed that RTHF decreased A549 cell adhesion in a dose-dependent manner. Cell invasion and migration were investigated using the Transwell assay and observed using an inverted microscope; the results showed that cell metastasis was significantly lower in the treatment group than that in the control group (P<0.01). Expression of metastasis-related matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) was detected by real-time polymerase chain reaction and Western blotting. The results showed that the expression of MMP-2, MMP-9, and TIMP-1 decreased, while that of TIMP-2 increased significantly in the RTHF group when compared with the results of the control group. These results show that RTHF exhibits antigrowth and antimetastasis activity in lung cancer A549 cells by decreasing the expression of MMP-2/-9 and TIMP-1 and increasing that of TIMP-2.

Essential hypertension treated by wuling powder and modified tianma gouteng decoction: a cohort study without controls.

OBJECTIVE: To evaluate the efficacy and safety of wuling powder and modified tianma gouteng decoction as an open add-on therapy for treating essential hypertension (EH). METHODS: 72 cases of EH patients took wuling powder and modified tianma gouteng decoction for 2 weeks to treat EH without changing their former medication. New York Heart Association (NYHA) heart function classification, heart rate, blood pressure and 6-min walking test were evaluated before and after treatment. RESULTS: Wuling powder and modified tianma gouteng decoction decreased the EH patients' heart rate, systolic pressure, diastolic pressure and increased the 6-min walking distance very significantly (P<0.01). The total effective rate on blood pressure was 91.7%, and the total effective rate on NYHA heart function improvement was 95.1%. No serious side effect appeared. CONCLUSION: From this study, treating EH with wuling powder and modified tianma gouteng decoction as an open add-on therapy seems to be effective and safe. Further randomized trials are needed to test the positive findings.

[Effects of soil compaction stress on the growth and nitrogen metabolism of cucumber roots].

Taking cucumber cultivar 'Jinchun 4' as test material, a pot experiment was conducted to study the effects of soil compaction stress on cucumber root growth, respiration rate, activity, and nitrogen metabolism at different development stages. Two treatment were installed, i. e., bulk density 1.25 g x cm(-3) (loose soil, CK) and 1.55 g x cm(-3) (compacted soil). Under the stress of soil compaction, the total root length, root surface area, root forks, and root tips at different development stages decreased significantly, the root elongation growth and lateral root emergence were constrained remarkably, but the root radial growth was stimulated and the root average diameter was significantly increased. Meanwhile, both the root activity and the respiration rate decreased significantly, the root NO3(-), free amino acids, and soluble protein contents had a large decrease, the root nitrate reductase, glutamine synthetase, and glutamate synthase activities decreased significantly, whereas the root NH4(+) content had a significant increase. All the results indicated that under the stress of soil compaction, the uptake of nitrate by cucumber roots decreased, the nitrogen assimilation was inhibited, and the nitrogen metabolism was remarkably constrained.

[Effects of soil compaction stress on respiratory metabolism of cucumber root].

A pot experiment with cucumber cultivar "Jingchun 4" was conducted to study the effects of soil compaction stress on the respiratory metabolism of cucumber root. Two treatments were installed, i.e. , soil bulk densities 1.20 and 1.55 g . cm-3. Under soil compaction stress, the activities of root pyruvate decarboxylase, alcohol dehydrogenase, and lactate dehydrogenase and the contents of root anaerobic respiration products alcohol, acetaldehyde, and lactate increased significantly, while the activities of the key enzymes involved in root aerobic respiration, including malate dehydrogenase, succinate dehydrogenase, and isocitrate dehydrogenase, decreased significantly, root pyruvate and succinate contents had significant increase, whereas root malate content decreased significantly. All the results illustrated that under soil compaction stress, the aerobic respiration of cucumber root was inhibited, while its anaerobic respiration was promoted.

[Study on treatment of iron-deficiency anemia by shengxuening].

OBJECTIVE: To observe the therapeutic effect of shengxuening (SXN) in treating iron-deficiency anemia (IDA) and to explore its molecular mechanism on iron metabolism balance regulation. METHODS: Patients with IDA were randomly divided into the treated group and the control group, 50 in each group. They were treated with SXN (0.1 g, three times per day) and ferrous gluconate (0.1 g, three times per day) respectively, for 30 days. Levels of serum iron (Fe), total iron binding capacity (TIBC), transferrin saturation (TS), serum ferritin (SF), transferrin (Tf), soluble transferrin receptor (sTfR) and blood routine test, as well as scoring of TCM qi-blood deficiency Syndrome were conducted before and after treatment. RESULTS: The total effective rate in the treated group reached 92%, it was shown that SXN could improve the iron metabolism, increase levels of Fe, TS, SF and reduce levels of TIBC, Tf, sTfR, it has obvious effect in promoting erythrocyte generation and could promote formation of leucocytes and platelets. The total effective rate in the control group was 32%, which was significantly lower than that in the treated group (P < 0.01). CONCLUSION: The effect of SXN in treating IDA and qi-blood deficiency Syndrome is evident, it could improve the iron metabolism, increase levels of Fe, TS, SF and lower levels of TIBC, Tf, sTfR.