Clinical signs, causes, and outcome of central cord syndrome in 22 cats.

OBJECTIVE: To describe the signalment, clinical findings, presumptive or definitive diagnosis, and outcome in cats with central cord syndrome (CCS). ANIMALS: 22 cats. CLINICAL PRESENTATION: Cats evaluated for CCS at 7 referral hospitals between 2017 and 2021 were included. Information retrieved from medical records included signalment, physical and neurological examination findings, diagnostic investigations, definitive or presumptive diagnosis, treatment, and follow-up. RESULTS: Median age at presentation was 9 years. Two neuroanatomical localizations were associated with CCS: C1-C5 spinal cord segments in 17 (77.3%) cats and C6-T2 spinal cord segments in 5 (22.7%) cats. Neuroanatomical localization did not correlate with lesion location on MRI in 8 (36.3%) cats. The most common lesion location within the vertebral column was over the C2 and C4 vertebral bodies in 6 (27.2%) and 5 (22.7%) cats, respectively. Peracute clinical signs were observed in 11 (50%) cats, acute in 1 (4.5%), subacute in 4 (18%), and chronic and progressive signs were seen in 6 (40.9%) cats. The most common peracute condition was ischemic myelopathy in 8 (36.3%) cats, whereas neoplasia was the most frequently identified chronic etiology occurring in 5 (22.7%) cats. Outcome was poor in 13 (59%) cats, consisting of 4 of 11 (36.6%) of the peracute cases, 3 of 4 (75%) of the subacute cases, and 6 of 6 of the chronic cases. CLINICAL RELEVANCE: Central cord syndrome can occur in cats with lesions in the C1-C5 and C6-T2 spinal cord segments. Multiple etiologies can cause CCS, most commonly, ischemic myelopathy and neoplasia. Prognosis depends on the etiology and onset of clinical signs.

Class III Antiarrhythmics and Periprocedural Torsades de Pointes.

This case series describes 2 women on prolonged therapy with class III antiarrhythmics who developed torsades de pointes polymorphic ventricular tachycardia in the setting of catheter ablation for atrial fibrillation as a result of QTc prolonging factors. Clinicians must exercise increased vigilance in the perioperative period in patients on QTc-prolonging medications. (Level of Difficulty: Intermediate.).

Primary Feline Tauopathy: Clinical, Morphological, Immunohistochemical, and Genetic Studies.

Tauopathies are a group of neurodegenerative diseases characterized by the pathological aggregation of hyperphosphorylated tau in neurons and glia. Primary tauopathies are not uncommon in humans but exceptional in other species. We evaluate the clinical, neuropathological, and genetic alterations related to tau pathology in 16 cats aged from 1 to 21 years with different clinical backgrounds. Interestingly, a 10-year-old female cat presented a six-year progressive history of mental status and gait abnormalities. The imaging study revealed generalized cortical atrophy. Due to the poor prognosis, the cat was euthanatized at the age of ten. Neuropathological lesions were characterized by massive neuronal loss with marked spongiosis and associated moderate reactive gliosis in the parietal cortex, being less severe in other areas of the cerebral cortex, and the loss of Purkinje cells of the cerebellum. Immunohistochemical methods revealed a 4R-tauopathy with granular pre-tangles in neurons and coiled bodies in oligodendrocytes. Deposits were recognized with several phospho-site antibodies (4Rtau, tau5, AT8, PFH, tau-P Thr181, tau-P-Ser 262, tau-P Ser 422) and associated with increased granular expression of active tau kinases (p38-P Thr180/Tyr182 and SAPK/JNK-P Thr138/Thr185). The genetic study revealed well-preserved coding regions of MAPT. No similar alterations related to tau pathology were found in the other 15 cats processed in parallel. To our knowledge, this is the first case reporting a primary 4R-tauopathy with severe cerebral and Purkinje cell degeneration in an adult cat with neurological signs starting at a young age.

The Prevalence of Depression Symptoms and Frequency of Risk Factors in Medical and Nursing Students: A Descriptive Cross-sectional Study.

OBJECTIVE: Currently, in Puerto Rico, there is a paucity of data regarding emotional health and depression in health professionals, specifically regarding trainees such as medical students and nursing students. The study intended to shed light on the prevalence of depression symptoms among medical and nursing students at a school of medicine in Puerto Rico. METHODS: In the fall of 2019, a descriptive cross-sectional study that included nursing and medical students in their first, second, and third years was performed. A survey consisting of the Patient Health Questionnaire (PHQ-9) and sociodemographic questions were used for data collection. Logistic regression analyses were used to determine the association of PHQ-9 scores and the risk factors linked to depression symptoms. RESULTS: A total of 173 (83.2%) out of 208 enrolled students participated in the study. Of the participants, 75.7% were medical students and 24.3% were nursing students. Of the risk factors studied, feelings of regret and lack of sleep were associated with a higher frequency of depression symptoms in medical students. For the nursing student population, suffering from a chronic disease was associated with a higher frequency of depression symptoms. CONCLUSION: Due to the increased risk of depression in healthcare professionals, identifying risk factors that can be addressed through early changes in behavior, or in institutional policies, is important in terms of working to mitigate the risk of mental health problems in this vulnerable population.

A Case of Culture-Negative Endocarditis Due to Bartonella henselae and Cat Scratch Disease.

This report describes the case of a 73-year-old female with a history of a prosthetic mitral valve and breast cancer who presented with fever and confusion. Brain imaging revealed multiple ischemic infarcts, and abdominal imaging demonstrated splenic infarcts. Workup with transesophageal echocardiogram revealed a vegetation on the aortic valve, but several blood cultures had no bacterial growth. Further history revealed a recent exposure to a stray cat, and the patient was found to have positive antibodies for Bartonella henselae, the organism implicated in cat scratch disease. She was treated with the appropriate course of antibiotics, and she made a full recovery to her baseline functional status. This report emphasizes the importance of a comprehensive patient history and a broad differential diagnosis.

Acute effects of a session of electroconvulsive therapy on brain-derived neurotrophic factor plasma levels.

Brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) are neurotrophins that play critical roles in brain neuronal function. Previous studies have established the association between BDNF and NGF signaling and severe mental disorders, but changes in BDNF plasma levels and electroconvulsive therapy (ECT) response are controversial. The aim of his study was to explore the acute effects of a single session of ECT on these neurotrophins signaling. Plasma levels of BDNF and NGF and their tyrosine kinase-type receptors expression in peripheral blood mononuclear cells (PBMCs) were determined before and two hours after a single ECT session in 30 subjects with a severe mental disorder. Two hours after an ECT session we found a statistically significant decrease of BDNF plasma levels (p=0.007). We did not find significant acute effects on NGF plasma levels or receptors expression in PBMCs. We found a significant inverse correlation between the time of convulsion and BDNF plasma levels decrease (r=-0.041, p=0.024). We have identified a decrease in BDNF plasma levels after 2h of a single ECT session. These results indicate the interest for future research in the role of neurotrophins in the response and safety of ECT.

Genetic deletion of the nuclear factor of activated T cells 5 in collecting duct principal cells causes nephrogenic diabetes insipidus.

Water homeostasis is tightly regulated by the kidneys via the process of urine concentration. During reduced water intake, the antidiuretic hormone arginine vasopressin (AVP) binds to the vasopressin receptor type II (V2R) in the kidney to enhance countercurrent multiplication and medullary osmolality, and increase water reabsorption via aquaporin-2 (AQP2) water channels. The importance of this AVP, V2R, and AQP2 axis is highlighted by low urine osmolality and polyuria in people with various water balance disorders, including nephrogenic diabetes insipidus (NDI). ELF5 and nuclear factor of activated T cells 5 (NFAT5) are two transcription factors proposed to regulate Aqp2 expression, but their role is poorly defined. Here we generated two novel mouse lines with principal cell (PC)-specific deletion of ELF5 or NFAT5 and phenotyped them in respect to renal water handling. ELF5-deficient mice (ELF5PC-KO ) had a very mild phenotype, with no clear differences in AQP2 abundance, and mild differences in renal water handling and maximal urinary concentrating capacity. In contrast, NFAT5 (NFAT5PC-KO ) mice had significantly higher water intake and their 24 h urine volume was almost 10-fold greater than controls. After challenging with dDAVP or 8 h water restriction, NFAT5PC-KO mice were unable to concentrate their urine, demonstrating that they suffer from NDI. The abundance of AQP2, other AQPs, and the urea transporter UT-A1 were greatly decreased in NFAT5PC-KO mice. In conclusion, NFAT5 is a major regulator of not only Aqp2 gene transcription, but also other genes important for water homeostasis and its absence leads to the development of NDI.

Central cord syndrome: clinical features, etiological diagnosis, and outcome in 74 dogs.

OBJECTIVE: To describe the clinical and neurologic signs, diagnostic investigations, definitive or presumptive diagnosis, treatment, and outcome of dogs presented with acute onset central cord syndrome (CCS). ANIMALS: 74 client-owned dogs evaluated for CCS at 5 referral hospitals between January 2016 and March 2021. PROCEDURES: Data were collected from the medical records of each dog, including patient signalment, physical and neurologic examination results, presence of signs of respiratory failure, diagnostic imaging findings, definitive or presumptive diagnosis, treatment and follow-up information. Descriptive statistics were calculated and bivariable analysis was performed to identify associations between selected variables. RESULTS: 2 neuroanatomic locations for the CCS were identified: C1-C5 spinal cord segments in 65 of 74 (88%) dogs and C6-T2 in 9 (12%) dogs. Neurolocalization did not correlate with the imaging findings in 43 (58%) dogs. Different diseases were associated with CCS. The most common condition was Hansen type I disk herniation in 27 (36%) dogs and hydrated nucleus pulposus extrusion in 16 (22%) dogs. Main lesion locations within the vertebral column associated with CCS were C3-C4 and C4-C5 intervertebral disk spaces in 21 (28%) and 18 (24%) dogs, respectively. Outcome was favorable in 69 (93%) dogs. Patients presenting with hypoventilation were 14.7 times more likely to have a poor outcome. CLINICAL RELEVANCE: CCS in dogs may be seen with lesions in the C1-C5 and C6-T2 spinal cord segments. Etiologies are variable. Total or partial improvement was achieved in most dogs with the appropriate treatment. Hypoventilation was associated with death.

Prevalence, distribution, and clinical associations of suspected postictal changes on brain magnetic resonance imaging in epileptic dogs.

OBJECTIVE: To determine the prevalence of presumed postictal changes (PC) on brain MRI in epileptic dogs, describe their distribution, and recognize possible correlations with different epilepsy features. ANIMALS: 540 client-owned dogs with epilepsy and a complete medical record that underwent brain MRI at 4 veterinary referral hospitals between 2016 and 2019. PROCEDURES: Data were collected regarding signalment, seizure type, seizure severity, time between last seizure and MRI, and etiological classification of epilepsy. Postictal changes were considered when solitary or multiple intraparenchymal hyperintense lesions were observed on T2-weighted and fluid-attenuated inversion recovery images and were hypointense or isointense on T1-weighted sequences, which were not confined to a vascular territory and showed no to mild mass effect and no to mild contrast enhancement. RESULTS: Sixty-seven dogs (12.4%) showed MRI features consistent with PC. The most common brain sites affected were the piriform lobe, hippocampus, temporal neocortex, and cingulate gyrus. Dogs having suffered cluster seizures or status epilepticus were associated with a higher probability of occurrence of PC, compared to dogs with self-limiting seizures (OR 2.39; 95% confidence interval, 1.33 to 4.30). Suspected PC were detected both in dogs with idiopathic epilepsy and in those with structural epilepsy. Dogs with unknown-origin epilepsy were more likely to have presumed PC than were dogs with structural (OR 0.15; 95% confidence interval, 0.06 to 0.33) or idiopathic epilepsy (OR 0.42; 95% confidence interval, 0.20 to 0.87). Time between last seizure and MRI was significantly shorter in dogs with PC. CLINICAL RELEVANCE: MRI lesions consistent with PC were common in epileptic dogs, and the brain distribution of these lesions varied. Occurrence of cluster seizures or status epilepticus, diagnosis of unknown origin epilepsy, and lower time from last seizure to MRI are predictors of suspected PC.

One-Pot Mechanochemical Synthesis of Mono- and Bis-Indolylquinones via Solvent-Free Multiple Bond-Forming Processes.

Bis-indolylquinones are fungal natural products endowed with interesting pharmacological properties. Most of the previously described methodologies in solution for the construction of the bis-indolylquinone framework show disadvantages associated with long reaction times and difficult, waste-generating purifications. A one-pot mechanochemical methodology for the synthesis of indolylquinones was developed, starting from indoles and dihaloquinones in the presence of FeCl3 or p-TsOH as catalysts and Fetizon's reagent as an oxidant. In contrast to solution chemistry, mechanochemical activation allowed the double addition of indole to a quinone substrate in one pot, leading to symmetrical or non-symmetrical bis-indolylquinones via a domino processes comprising up to six steps. In terms of sustainability, the method has several advantages over the solution protocol, including much shorter reaction times, no external heating, one-pot operation, and the absence of chromatography, leading to a drastically better performance in green metrics and demonstrating the application of several principles of green chemistry, in particular principles 2, 3, and 5.

Multidisciplinary approach in the treatment of tendinous foot involvement in rheumatoid arthritis.

INTRODUCTION: Patients with rheumatoid arthritis frequently consult for pain resulting from involvement of the tendons of the foot. This pain negatively affects foot biomechanics and quality of life. The most widely used treatment option for this condition is ultrasound-guided steroid injection, while other treatments were recommended such as heel pad, splints, and footwear. OBJECTIVE: To evaluate a joint intervention (rheumatology and podiatry) comprising an orthotic-podiatric treatment and infiltrations. We evaluated the response using ultrasound monitoring, a pain scale, functional tests, and assessment of patient satisfaction. METHODS: We performed a non-controlled blinded prospective interventional study of 96 patients with foot pain and selected those with ultrasound-confirmed tendon involvement. Patients enrolled started intervention treatment and were followed for 6 months. The outcome of the intervention was compared with the patient's baseline status. The pre-post differences in the secondary variables (pain, disability) were analyzed using the t test and contingency tables or the Mann-Whitney test. RESULTS: Using our protocol, we recorded a rapid and significant reduction in the intensity of pain, in the foot function index, and in the ultrasound parameters (grayscale and Doppler). Structural damage to the tendon improved more slowly, with significant outcomes only at the last visit with respect to baseline. Abnormal foot support was detected in 50% of patients, and 79.5% were using inappropriate footwear. CONCLUSIONS: Our multidisciplinary therapeutic protocol enabled a very significant improvement in tendon involvement. It was well-tolerated, with a high degree of satisfaction, and was easily evaluated using ultrasound. No changes in background medication were necessary. Key Points • Multidisciplinary evaluation of patients with RA is advisable because it improves the treatment management in cases of inflammatory activity and structural abnormalities of the foot. • Comprising orthopedic-podiatric treatment (heel, splints, and suitable footwear) and infiltrations, in terms of clinical, ultrasound, and functional recovery of the foot tendons. • The therapy protocol we propose led to a significant improvement in pain relief and functional recovery.

High dietary potassium causes ubiquitin-dependent degradation of the kidney sodium-chloride cotransporter.

The thiazide-sensitive sodium-chloride cotransporter (NCC) in the renal distal convoluted tubule (DCT) plays a critical role in regulating blood pressure (BP) and K+ homeostasis. During hyperkalemia, reduced NCC phosphorylation and total NCC abundance facilitate downstream electrogenic K+ secretion and BP reduction. However, the mechanism for the K+-dependent reduction in total NCC levels is unknown. Here, we show that NCC levels were reduced in ex vivo renal tubules incubated in a high-K+ medium for 24-48 h. This reduction was independent of NCC transcription, but was prevented using inhibitors of the proteasome (MG132) or lysosome (chloroquine). Ex vivo, high K+ increased NCC ubiquitylation, but inhibition of the ubiquitin conjugation pathway prevented the high K+-mediated reduction in NCC protein. In tubules incubated in high K+ media ex vivo or in the renal cortex of mice fed a high K+ diet for 4 days, the abundance and phosphorylation of heat shock protein 70 (Hsp70), a key regulator of ubiquitin-dependent protein degradation and protein folding, were decreased. Conversely, in similar samples the expression of PP1α, known to dephosphorylate Hsp70, was also increased. NCC coimmunoprecipitated with Hsp70 and PP1α, and inhibiting their actions prevented the high K+-mediated reduction in total NCC levels. In conclusion, we show that hyperkalemia drives NCC ubiquitylation and degradation via a PP1α-dependent process facilitated by Hsp70. This mechanism facilitates K+-dependent reductions in NCC to protect plasma K+ homeostasis and potentially reduces BP.

Doxycycline Significantly Enhances Induction of Induced Pluripotent Stem Cells to Endoderm by Enhancing Survival Through Protein Kinase B Phosphorylation.

BACKGROUND AND AIMS: Induced pluripotent stem cells (iPSCs) provide an important tool for the generation of patient-derived cells, including hepatocyte-like cells, by developmental cues through an endoderm intermediate. However, most iPSC lines fail to differentiate into endoderm, with induction resulting in apoptosis. APPROACH AND RESULTS: To address this issue, we built upon published methods to develop an improved protocol. We discovered that doxycycline dramatically enhances the efficiency of iPSCs to endoderm differentiation by inhibiting apoptosis and promoting proliferation through the protein kinase B pathway. We tested this protocol in >70 iPSC lines, 90% of which consistently formed complete sheets of endoderm. Endoderm generated by our method achieves similar transcriptomic profiles, expression of endoderm protein markers, and the ability to be further differentiated to downstream lineages. CONCLUSIONS: Furthermore, this method achieves a 4-fold increase in endoderm cell number and will accelerate studies of human diseases in vitro and facilitate the expansion of iPSC-derived cells for transplantation studies.

Common and external iliac artery occlusion in Behçet's disease: a case of anchoring bias.

This report describes the case of a 45-year-old woman with a history of Behçet's disease and complex regional pain syndrome of her legs who presented with severe pain and swelling in her left lower extremity. The patient was initially diagnosed with exacerbation of complex regional pain syndrome, which fit the symptom complex of hyperalgesia, oedema and skin temperature changes. However, after unsuccessful attempts at significant pain relief during admission, CT angiography demonstrated occlusion of the left common and external iliac arteries, a limb-threatening emergency. This case describes an example of anchoring bias, a type of cognitive bias in which there is a tendency to rely too heavily on an initial piece of information, the 'anchor', when making decisions. This report emphasises that clinicians should be aware of biases when making decisions and avoid anchoring bias by asking themselves if their diagnosis is influenced by any leading pieces of information.

The Correlation of English Language Proficiency and Indices of Stress and Anxiety in Migrants from Puerto Rico after Hurricane Maria: A Preliminary Study.

OBJECTIVE: Using validated psychological assessment instruments, this study examined the psychological distress associated with potential language barriers experienced by over 135 000 Puerto Rican residents who either temporarily or permanently migrated to the continental United States with the landfall of Hurricane Maria in 2017. METHODS: Participants were Puerto Rican residents (n = 107) who remained in Puerto Rico (control) or left the island for at least 3 months because of Hurricane Maria (migrants). Participants completed an online survey in their preferred language (Spanish or English), which assessed self-reported English language proficiency, Kessler Psychological Distress Scale (K6), Posttraumatic Stress Disorder Checklist for DSM 5, Patient Health Questionnaire 9-item depression scale, and the Generalized Anxiety Disorder 7-item scale. It was hypothesized that migrants with lower self-reported English proficiency would have comparatively higher indices of post-disaster distress than those with a higher proficiency. RESULTS: Dividing the migrant group by preferred language for questionnaire completion, the Fisher's exact test showed significant differences in prevalence of severe mental distress, as defined by K6 scores above 13, between the Spanish-preferring migrants (30.4%), English-preferring migrants (0%), and controls (9.6%). CONCLUSION: Our results support a possible correlation between decreased language proficiency in post-disaster migrants and a higher risk factor for severe mental distress.

Rapid Aldosterone-Mediated Signaling in the DCT Increases Activity of the Thiazide-Sensitive NaCl Cotransporter.

BACKGROUND: The NaCl cotransporter NCC in the kidney distal convoluted tubule (DCT) regulates urinary NaCl excretion and BP. Aldosterone increases NaCl reabsorption via NCC over the long-term by altering gene expression. But the acute effects of aldosterone in the DCT are less well understood. METHODS: Proteomics, bioinformatics, and cell biology approaches were combined with animal models and gene-targeted mice. RESULTS: Aldosterone significantly increases NCC activity within minutes in vivo or ex vivo. These effects were independent of transcription and translation, but were absent in the presence of high potassium. In vitro, aldosterone rapidly increased intracellular cAMP and inositol phosphate accumulation, and altered phosphorylation of various kinases/kinase substrates within the MAPK/ERK, PI3K/AKT, and cAMP/PKA pathways. Inhibiting GPR30, a membrane-associated receptor, limited aldosterone's effects on NCC activity ex vivo, and NCC phosphorylation was reduced in GPR30 knockout mice. Phosphoproteomics, network analysis, and in vitro studies determined that aldosterone activates EGFR-dependent signaling. The EGFR immunolocalized to the DCT and EGFR tyrosine kinase inhibition decreased NCC activity ex vivo and in vivo. CONCLUSIONS: Aldosterone acutely activates NCC to modulate renal NaCl excretion.

Post-Hurricane Distress Scale (PHDS): A Novel Tool for First Responders and Disaster Researchers.

OBJECTIVE: The aim of this study was the construction and validation of a novel research instrument to quantify the degree of post-hurricane trauma and distress in an affected population. The Post-Hurricane Distress Scale (PHDS) has quantitative measures of both acute and prolonged distress, attributable to meteorological and hydrological disasters. METHODS: A careful evaluation of existing questionnaires, as well as extensive canvasing of the post-Maria population of Puerto Rico, availed the construction of the PHDS. The PHDS consists of 20 items, organized into 4 subscales. The PHDS was pre-validated (n=79), revised, and then distributed to a broad sampling of the post-Hurricane Maria Puerto Rican population (n=597). Validation, including factor analysis, analyses of concurrent validity, discriminant validity, and internal reliability, was performed. RESULTS: After comparing various scales, factor loading profiles, concurrent validities, and models of fit, we show that the PHDS is best scored as a single 0-6 distress scale. When compared with the Traumatic Exposure Severity Scale, the PHDS shows superior concurrent validity, more accurately predicting scores for the Peritraumatic Distress Inventory, Impact of Event Scale - Revised, and Generalized Anxiety Disorder 7 Scale. The PHDS shows good internal reliability and discriminant validity. CONCLUSIONS: The PHDS represents a novel, useful instrument for disaster first-responders and researchers. The prompt identification of high-risk populations is possible using this instrument. (Disaster Med Public Health Preparedness. 2019;13:82-89).

Long-term follow-up of surgical resection alone for primary intracranial rostrotentorial tumors in dogs: 29 cases (2002-2013).

Intracranial neoplasia is frequently encountered in dogs. After a presumptive diagnosis of intracranial neoplasia is established based on history, clinical signs and advanced imaging characteristics, the decision to treat and which treatment to choose must be considered. The objective of this study is to report survival times (ST) for dogs with intracranial meningiomas and gliomas treated with surgical resection alone (SRA), to identify potential prognostic factors affecting survival, and to compare the results with the available literature. Medical records of 29 dogs with histopathologic confirmation of intracranial meningiomas and gliomas treated with SRA were retrospectively reviewed. For each dog, signalment, clinical signs, imaging findings, type of surgery, treatment, histological evaluation, and ST were obtained. Twenty-nine dogs with a histological diagnosis who survived >7 days after surgery were included. There were 15 (52%) meningiomas and 14 (48%) gliomas. All tumors had a rostrotentorial location. At the time of the statistical analysis, only two dogs were alive. Median ST for meningiomas was 422 days (mean, 731 days; range, 10-2735 days). Median ST for gliomas was 66 days (mean, 117 days; range, 10-730 days). Kaplan-Meier analysis indicated that ST was significantly longer for meningiomas than for gliomas (P<0.05). A negative correlation between the presence of a midline shift and ST (P=0.037) and ventricular compression and ST (P=0.038) was observed for meningiomas. For gliomas, there were no significant associations between ST and any of the variables evaluated. In conclusion, the results of this study suggest that, for dogs that survived >7 days postoperatively, SRA might be an appropriate treatment, particularly for meningiomas, when radiation therapy is not readily available. Also, the presence of midline shift and ventricular compression might be negative prognostic factors for dogs with meningiomas.

Experimental Evaluation of Proposed Small-Molecule Inhibitors of Water Channel Aquaporin-1.

The aquaporin-1 (AQP1) water channel is a potentially important drug target, as AQP1 inhibition is predicted to have therapeutic action in edema, tumor growth, glaucoma, and other conditions. Here, we measured the AQP1 inhibition efficacy of 12 putative small-molecule AQP1 inhibitors reported in six recent studies, and one AQP1 activator. Osmotic water permeability was measured by stopped-flow light scattering in human and rat erythrocytes that natively express AQP1, in hemoglobin-free membrane vesicles from rat and human erythrocytes, and in plasma membrane vesicles isolated from AQP1-transfected Chinese hamster ovary cell cultures. As a positive control, 0.3 mM HgCl2 inhibited AQP1 water permeability by >95%. We found that none of the tested compounds at 50 µM significantly inhibited or increased AQP1 water permeability in these assays. Identification of AQP1 inhibitors remains an important priority.

Disruption of ku70 involved in non-homologous end-joining facilitates homologous recombination but increases temperature sensitivity in the phytopathogenic fungus Penicillium digitatum.

The dominant mechanism to repair double-stranded DNA breaks in filamentous fungi is the non-homologous end joining (NHEJ) pathway, and not the homologous recombination (HR) pathway that operates in the mutation of genes by replacement of target DNA for selection cassettes. The key to improve HR frequency is the inactivation of the NHEJ pathway by eliminating components of its Ku70/80 heterodimeric complex. We have obtained ku70 mutants of Penicillium digitatum, the main citrus postharvest pathogen. The increased efficiency of HR in Δku70 strains was demonstrated by the generation of mutants in two different chitin synthase genes (PdchsII and PdchsV). P. digitatum Δku70 strains showed no differences from the parental strain in vegetative growth, asexual development or virulence to citrus fruit, when experiments were conducted at the optimal temperature of 24°C. However, growth of Δku70 strains at temperatures higher than 24°C demonstrated a detrimental effect in axenic growth and conidia production. These observations are in agreement with previous studies describing differences between ku70 mutants and their parental strains in some fungal species, and must be taken into account for future applications of the Δku approach to increase HR efficiency in fungi.