First report of areolate mildew of cotton, caused by Ramulariopsis pseudoglycines in Mississippi.

In August of 2022, cotton (Gossypium hirsutum L.) growing in several north central Mississippi counties was observed to exhibit yellowish lesions on the adaxial leaf surface with white powdery fungal growth on the corresponding abaxial surface. By the end of the 2022 growing season, 19 Mississippi counties were observed to have infected cotton. Symptomatic leaves were collected from affected plants, sealed in plastic freezer bags, stored on ice in a cooler, and transported to the laboratory. Prior to isolation, the pathogen was microscopically examined and determied to be morphologically similar to the description of Ramulariopsis spp. (Ehrlich and Wolf 1932). Using a sterile needle, conidia were transferred to V8 medium amended with chloramphenicol (75 mg/liter) and streptomycin sulfate (125 mg/liter) and incubated in the dark at 25°C. After 14 days, the colony diameter was measured, and morphological characteristics were consistent with previous descriptions (Videira et al. 2016; Volponi et al. 2014). On V8 medium, the 7 mm diameter colonies grew raised, lumpy, and lobed, with iron-grey coloration. The mycelia were hyaline, septate, branched, and 1 to 3 µm in diameter. Conidia ranged from 2.8 to 25.6 µm in length and 1.0 to 4.9 µm in width (x = 12.8 × 3.1 µm; n = 20). Pure cultures were obtained on V8 medium, and DNA was extracted from a 14-day-old-culture. TW098-22, a representative isolate, was amplified, and sequenced targeting the internal transcribed spacer (ITS), translation elongation factor 1-α (TEF 1-α), and actin (ACT) genes as described by Videira et al. (2016). The consensus sequences were deposited in GenBank (accession no. OQ653427, OR157986, OR157987). BLASTn query of the NCBI GenBank showed 100% identity of the 483-bp (ITS) and 706-bp TEF 1-α sequences from TW098-22 with Ramulariopsis pseudoglycines CPC 18242 (type culture; Videira et al. 2016). Koch's postulates were performed after multiplying individual colonies by streaking on V8 medium as above. Culture plates were subsequently incubated at 25°C for 14 days in the dark. Colonies were aseptically transferred to 50 ml centrifuge tubes containing 50 ml of autoclaved reverse osmosis (RO) water amended with Tween 20 (0.01%). The resulting inoculum suspension was adjusted to 13.5 × 105 conidia/ml using a hemocytometer. The foliage of five, 25-day-old cotton plants were sprayed with 10 ml of the suspension and a plastic bag was placed over each plant to maintain humidity for 30 days. Five plants were sprayed with sterilized RO water to serve as controls. Plants were incubated in a growth chamber at 25°C and ~70% relative humidity with 16:8 h of light:dark. Thirty days post-inoculation, foliar symptoms and signs were observed on all inoculated plants including small necrotic lesions and white powdery growth. Control plants remained asymptomatic. The trial was repeated. When reisolated, the colony and conidia morphology and DNA sequence (ITS) were consistent with the description of the original field isolate. Areolate mildew of cotton can be caused by two species of Ramulariopsis: R. gossypii and R. pseudoglycines (Videira et al. 2016). The two species have been reported in Brazil (Mathioni et al. 2021); however, this is the first report of R. pseudoglycines in the United States. In addition, even though areolate mildew has previously been reported from much of the southeastern U.S. (Anonymous 1960), the report herein serves as the first description of R. pseudoglycines in Mississippi and U.S. cotton.

Falciform ligament torsion as a rare aetiology of the acute abdomen.

The falciform ligament is a remnant of the embryonic ventral mesentery containing the obliterated umbilical vein and round ligament. It extends from the umbilicus to the superior aspect of the diaphragm. We report about a 53-year-old fit and well patient who presented with acute upper abdominal pain with tenderness to palpation. Ultrasound scan was unremarkable, but blood tests revealed raised inflammatory markers. Thus, computed tomography was performed. This demonstrated acute torsion and fat necrosis of the falciform ligament, which was the aetiology of the upper abdominal pain. Such pathology is rare with 23 previously reported cases. Conservative management is usually proposed, but on occasion, surgical intervention may be warranted in cases that do not respond to initial supportive measures. We describe this case to demonstrate a rare cause of a common presentation to the surgical service.

Loin Pain Hematuria Syndrome-a case report.

Loin pain hematuria syndrome (LPHS) is a rare idiopathic condition. LPHS can present with both unilateral and bilateral loin pain, microscopic or macroscopic hematuria. It is a diagnosis of exclusion. The management options for this condition include pain management with narcotics or opioids, renal denervation, kidney autotransplantation and neurectomy or nephrectomy. However, these treatment modalities are the last resort.

Meta-analysis and trial sequential analysis of randomized evidence comparing general anesthesia vs regional anesthesia for laparoscopic cholecystectomy.

BACKGROUND: In an effort to further reduce the morbidity and mortality profile of laparoscopic cholecystectomy, the outcomes of such procedure under regional anesthesia (RA) have been evaluated. In the context of cholecystectomy, combining a minimally invasive surgical procedure with a minimally invasive anesthetic technique can potentially be associated with less postoperative pain and earlier ambulation. AIM: To evaluate comparative outcomes of RA and general anesthesia (GA) in patients undergoing laparoscopic cholecystectomy. METHODS: A comprehensive systematic review of randomized controlled trials with subsequent meta-analysis and trial sequential analysis of outcomes were conducted in line with Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement standards. RESULTS: Thirteen randomized controlled trials enrolling 1111 patients were included. The study populations in the RA and GA groups were of comparable age (P = 0.41), gender (P = 0.98) and body mass index (P = 0.24). The conversion rate from RA to GA was 2.3%. RA was associated with significantly less postoperative pain at 4 h [mean difference (MD): - 2.22, P < 0.00001], 8 h (MD: -1.53, P = 0.0006), 12 h (MD: -2.08, P < 0.00001), and 24 h (MD: -0.90, P < 0.00001) compared to GA. Moreover, it was associated with significantly lower rate of nausea and vomiting [risk ratio (RR): 0.40, P < 0.0001]. However, RA significantly increased postoperative headaches (RR: 4.69, P = 0.03), and urinary retention (RR: 2.73, P = 0.03). The trial sequential analysis demonstrated that the meta-analysis was conclusive for most outcomes, with the exception of a risk of type 1 error for headache and urinary retention and a risk of type 2 error for total procedure time. CONCLUSION: Our findings indicate that RA may be an attractive anesthetic modality for day-case laparoscopic cholecystectomy considering its associated lower postoperative pain and nausea and vomiting compared to GA. However, its associated risk of urinary retention and headache and lack of knowledge on its impact on procedure-related outcomes do not justify using RA as the first line anesthetic choice for laparoscopic cholecystectomy.

Acute appendicitis located within Amyand's hernia-a complex case with concurrent acute cholecystitis.

Amyand's hernia is the presence of the vermiform appendix within an inguinal hernia sac. It is rare, and even rarer is the presence of acute appendicitis within the sac. It presents in a variety of different ways and often is only diagnosed intra-operatively. We present the case of a 90 year old male with extensive co-morbidities presenting with right upper quadrant pain, who on computed tomography scan of the abdomen, had acute cholecystitis alongside acute appendicitis within Amyand's hernia. Ultimately given his co-morbidities, a conservative approach with prolonged antibiotic therapy was adopted, with a successful outcome. This case highlights that although classifications for treatment of Amyand's hernia exist, careful clinical assessment is warranted in each case to ensure optimal outcome based upon individual circumstances.

Septic arthritis of the manubriosternal joint presenting as a chest-wall swelling in an immunocompetent patient.

Septic arthritis of the manubriosternal joint is a rare pathology, often mistaken for other disease processes given its location and chest pain symptoms. We present a case of a 58-year-old man presenting with a dull ache in his chest after returning from a holiday. Initially under the care of the physicians locally, he was investigated for respiratory and cardiac causes of chest pain with no findings. Eventually, a lump was noted on examination of the chest prompting further imaging, which confirmed a diagnosis of manubriosternal septic arthritis. He was discussed and referred to tertiary cardiothoracics, who recommended conservative treatment with 6 weeks of antibiotics. To date, there has been a good recovery. We present this case alongside a discussion of the limited literature, in particular highlighting how difficult a diagnosis it is to make but one that surgeons and medics alike should be aware of.

De Garengeot's hernia with appendicitis-a rare cause of an acutely painful groin swelling.

De Garengeot hernia is a rare subtype of femoral hernia whereby the vermiform appendix is located within the hernial sac. Even rarer is the presence of appendicitis within the hernia sac. De Garengeot's hernia is difficult to diagnose pre-operatively and can prove technically difficult at operation particularly with regards to mobilization of the caecum and appendix in order to perform appendicectomy. Laparoscopic, open, with and without mesh repair of de Garengeot hernia have all been described in the literature with varying degrees of success. We present a case of an 82 year old lady presenting with an acutely painful right sided groin lump. CT scan revealed the presence of de Garengeot hernia with acute appendicitis. We describe in text and photo format our approach to the hernia repair, appendicectomy and provide a short review of the literature with regards to the different operative approaches to such a patient.

Unusual presentation of acute pancreatitis following high tibial osteotomy.

We report a case of a 55-year-old man taxi driver admitted electively for high tibial osteotomy for relief of bilateral medial compartment osteoarthritis. He was obese and an ex-smoker but medical history was otherwise unremarkable. Day 1 postoperatively the patient started to suffer vomiting episodes and the abdomen was distended. C Reactive protein and white cell count were both elevated but other bloods were normal. CT abdomen and pelvis was performed which showed acute interstitial pancreatitis. Glasgow score was calculated as 2. He was treated aggressively with analgesia, intravenous fluid and intravenous antibiotics. He has made a good recovery following these conservative measures. To our knowledge, this is the first case in the literature reporting pancreatitis as a complication of high tibial osteotomy. This case highlights the importance of close monitoring for abdominal complications even in the setting of elective orthopaedic surgery where it is a rare phenomenon.

A study of fecal coliform sources at a coastal site using colored dissolved organic matter (CDOM) as a water source tracer.

Optical properties of colored dissolved organic matter (CDOM) were measured as a tracer of polluted waters in a Southern California surf-zone with consistently high levels of fecal indicator bacteria. Salinity, temperature, fecal coliform, absorbance (200-700nm) and fluorescence (lambda(excitation)=350nm; lambda(emission)=360-650nm) were measured in the creek and surf-zone during a dry and rain event. Fluorescence to absorption ratios for CDOM were used to distinguish water masses, with two distinct CDOM end-members identified as creek (flu/abs=8.7+/-0.8x10(4)) and coastal (flu/abs=2.2+/-0.3x10(4)). Waters containing the same CDOM end-member had highly variable bacterial levels during the dry event, suggesting intermittent sources of bacteria added to a uniform water source, consistent with marine birds. During the rain event, increased levels of the creek end-member and bacteria indicated a second bacteria source from runoff.

Insulin capture by an insulin-linked polymorphic region G-quadruplex DNA oligonucleotide.

Insulin capture by a G-quadruplex DNA oligonucleotide containing a two-repeat sequence of the insulin-linked polymorphic region (ILPR) of the human insulin gene promoter region is reported. The immobilized oligonucleotide was demonstrated to capture human insulin from standard solutions and from nuclear extracts of pancreatic cells with high selectivity, using affinity MALDI mass spectrometry and affinity capillary chromatography. Insulin was preferentially captured by the two-repeat ILPR oligonucleotide over another G-quadruplex-forming oligonucleotide, the thrombin-binding aptamer, as well as over a single repeat of the ILPR sequence that is not capable of forming the G-quadruplex architecture. Binding was shown to involve the beta chain of insulin. The discovery raises the possibility that insulin may bind to G-quadruplex DNA formed in the ILPR in vivo and thereby play a role in modulation of insulin gene expression, and it provides a basis for design of insulin analogues to probe this hypothesis. The availability of a DNA ligand to human insulin has analytical importance as well, offering an alternative to antibodies for in vitro or in vivo detection and sensing of insulin as well as its isolation and purification from biological samples.

Aptamer stationary phase for protein capture in affinity capillary chromatography.

The thrombin-binding DNA aptamer was used with thrombin as a model system to investigate protein capture using aptamer stationary phases in affinity capillary chromatography. The aptamer was covalently attached to the inner surface of a bare fused-silica glass capillary to serve as the stationary phase. Proteins were loaded onto the capillary via an applied pressure. The capillary was then washed to remove unbound and non-specifically associated proteins. Finally, the bound protein was released and eluted using 20 mM Tris buffer containing 8 M urea, pH 7.3, at 50 degrees C. Eluate was collected after each step (load, wash and elute) and relative amounts of protein each were compared using fluorescence spectroscopy. The identity of the protein in the collections was confirmed using matrix assisted laser desorption ionization-time-of-flight (MALDI-TOF) mass spectrometry. The experiment was repeated for thrombin on a bare (unmodified) capillary and a capillary coated with a scrambled-sequence, non-G-quartet forming oligonucleotide that does not bind with thrombin. The results show that the aptamer stationary phase captures approximately three times as much thrombin as the control columns. The experiment was also repeated using human serum albumin (HSA) alone and in an equimolar mixture with thrombin. HSA was not retained on the aptamer capillary, nor did it affect the capture of thrombin from the mixture.

Molecular ontogeny of the human antibody repertoire to the Haemophilus influenzae type B polysaccharide: expression of canonical variable regions and their variants in vaccinated infants.

A structurally conserved antibody combining site, encoded by the IGH V3-23 and kappa A2 variable (V) region gene segments, predominates the adult immune response to the Haemophilus influenzae type b (Hib) capsular polysaccharide (PS). This site has been elevated to canonical status based upon its relative molecular uniformity and prevalence in adults. To date, no studies have examined the primary structure of Hib PS-specific antibodies in young infants, who are the primary targets of Hib vaccination. In this study we show that canonical Hib PS-specific heavy (H) and light (L) chain V regions are present in 4-month-old infants following two vaccinations with Hib PS-protein conjugates. The infant V regions contain sequence polymorphisms that resemble those found in adult antibodies, as well as polymorphisms at position 95a of the A2 L chain not previously observed in adults. In vitro studies of Fab fragments and recombinant IgG2 antibodies using these V regions identify sequence polymorphisms that impact Hib PS binding affinity and bactericidal activity. These results demonstrate the establishment of canonical V regions in early ontogeny and provide a structural explanation of how canonical antibodies in the infant can vary in their affinity and protective activity against Hib.