Pancreatic Myeloid Sarcoma Causing Obstructive Jaundice: A Case Report and Literature Review.

Myeloid sarcoma (MS) is an extramedullary manifestation of acute myeloid leukemia (AML) and commonly occurs in sites such as the lymph nodes, skin, soft tissues, and bone. It more rarely manifests in the pancreas, with less than 20 cases reported in the literature since 1987. Despite its rarity, MS should be considered in the differential diagnosis of a soft tissue mass causing obstructive jaundice, especially if the patient has a known hematologic disease. Isolated cases of pancreatic MS have been known to progress to AML; therefore, it is crucial to differentiate MS from more common diagnoses, such as pancreatic cancer or pancreatitis. This is a case of a 70-year-old male with symptomatic obstructive jaundice secondary to pancreatic MS, ultimately requiring endoscopic ultrasound (EUS) and endoscopic retrograde cholangiopancreatography (ERCP) for diagnosis and management. Also included is a comprehensive review of previous case reports with similar clinical presentations, management, and treatment of pancreatic MS.

Reconstruction of Head and Neck Defects Using Medial Sural Artery Perforator Free Flap.

AIM: To study the use of medial sural artery perforator (MSAP) free flap in head and neck reconstruction. MATERIAL AND METHOD: This was a prospective study. The patients with cancers of head and neck underwent excision of tumor along with neck dissection, and MSAP free flap was used for reconstruction. RESULTS: The free MSAP flaps were used in 30 patients to reconstruct head and neck soft tissue defects. There were sixteen male and six female patients with the median age of 40 years. The most common site of tumor resection was the tongue (14 cases), followed by buccal mucosa (12 cases), neck skin(2 cases), skin over parotid(1 case) and lip(1 case). Average flap size was 56 cm2. Thickness of the flap ranged from 4 to 8 mm. The length of the vascular pedicle ranged from 8 to 14 cm (12 cm mean) which provides sufficient length during vessel anastomosis. Arterial diameter ranged from 1.0 to 1.5 mm(Average - 1.25 mm) and venous diameter of both veins in pedicle ranged from 1.5 to 2.5 mm(Average - 2 mm) in size. Most flaps were based on two perforators. Primary closure was attained in 11 cases whereas 19 patients required split thickness skin graft(STSG). The average flap harvesting time was 45 min. Flap was failed in two cases. CONCLUSION: MSAP is good alternative to FRAFF in the reconstruction of defect after resection of head and neck cancer.

Imaging spectrum, associations and outcomes in acute invasive fungal rhino-ocular-cerebral sinusitis in patients with COVID-19 pneumonia.

Background: Coronavirus Disease 2019 (COVID-19) has been speculated to enhance mucormycosis infection due to its immune-altering pathophysiology. Early identification of high-morbidity conditions is crucial for optimal treatment and improved outcomes. Methods: A retrospective study was conducted on 63 patients with clinical and microbiological evidence of rhino-ocular-cerebral mucormycosis, who had a history of COVID-19 infection. The clinical, demographic, and imaging data were retrieved and analyzed. Descriptive statistics (mean [SD] and frequency [%]) were used to describe important characteristics across audit cycles. Results: Out of 63 patients, 54 (85.71%) patients had associated comorbidities, with diabetes mellitus being common comorbidity and all patients had received injectable and/or oral corticosteroids. Imaging showed nasal and paranasal sinus, perisinus, maxillary alveolar arch, and hard palate involvement in 62 (98.41%), 33 (52.38%), 5 (7.94%) and 5 (7.94%) patients, respectively. Orbital involvement was seen in 24 (38.10%) patients. Skull base involvement was seen in 11 (17.46%) patients, and intracranial extension of disease was present in 11 (17.46%) patients. A total of 16 patients were on mechanical ventilation, of whom 3 succumbed. The mean (standard deviation [SD]) intensive care unit (ICU) stay was 13.2 days (6.8) for 5 patients who succumbed and 6.4 days (4.6) for 30 patients who survived (P value = 0.008). Conclusion: Cross-sectional imaging not only provides the extent of disease spread but also plays a vital role in providing a surgical roadmap to treating surgeons and in predicting prognosis in patients with invasive fungal infections.

Application of Belfast 15/30 dB Rule of Thumb in Assessing Hearing Benefit in Middle Ear Reconstructive Surgery.

The objective of this study is to assess the concordance between Belfast 15/30 dB rule of thumb and subjective hearing benefit in middle ear reconstruction surgery. A total of 105 cases of chronic otitis media (COM) (both mucosal and squamosal type) with conductive hearing loss and who underwent middle ear reconstructive surgery during the study period from January 2019 to January 2022 were included. All cases were followed up at 6 week and at 3 months of postoperative duration for subjective assessment of hearing with questionnaire and pure tone audiometry as per standard of care and correlated with Belfast 15/30 dB rule of thumb. Concordance with Belfast rule of thumb was seen in 73.4% cases of both mucosal and squamosal type of COM. Belfast 15/30 dB rule of thumb is a valuable and useful tool to predict hearing benefit following middle ear reconstructive surgery, but, it suffers from the disadvantages of all or none phenomenon and it does not grade the level of hearing improvement. The prediction of postoperative subjective hearing with this Belfast rule of thumb is more reliable in mucosal type of COM than in squamosal type of COM.

Functional Reconstruction of Through and Through Cheek Defect with Free Composite Bi-pedalled Anterolateral Thigh Flap.

Extensive composite defects involving the lip and cheek present difficult reconstructive challenges in view of functional recovery like oral competence, articulation, speech and mastication. This study presents our results of reconstructing through and through cheek defect with the use of free anterolateral thigh (ALT) flaps. All the patients with carcinoma of buccal mucosa and cheek who underwent through and through cheek resection and reconstructed with free composite pedalled anterolateral thigh flap between March 2019 to March 2020 were included in the study. We have assessed the post-operative functional outcome using University of Washington Quality of Life Questionnaire at the end of 12 months duration. We have excluded lost to follow-up and flap failure patients. Fifty patients were matched our inclusion criteria. Mean age was 43 years. Male to female ratio was 4.6:1. Stage 3 disease was in 8 patients and stage 4 disease was in 42 patients. Average size of the flap required was 15 × 7.5 cm, maximum was 24 × 11 cm. Oral competence was good in 74% patients. 80% patients were able to take semisolid diet, 8% were on liquid diet and 12% were dependent on RT feed. Average physical function (70.47 ± 19.09), social-emotional sub-score (81.72 ± 16.63) and composite scores (76.09 ± 17.86) were good and within acceptable range. Chewing (53 ± 29.29) and swallowing (64 ± 21.57) scored poorly among 12 domains. For extensive through and through cheek defects bipedelled ALT provides good functional outcome. In view of advance disease status extensive resection would be responsible for bone and mucosal loss affecting chewing and swallowing. Supplementary Information: The online version contains supplementary material available at 10.1007/s12070-022-03416-3.

Pancreatic Cancer Screening for At-Risk Individuals (Pancreas Scan Study): Yield, Harms, and Outcomes From a Prospective Multicenter Study.

INTRODUCTION: Guidelines endorse pancreatic cancer screening in genetically susceptible individuals. We conducted a prospective, multicenter study to determine yield, harms, and outcomes of pancreatic cancer screening. METHODS: All high-risk individuals undergoing pancreatic cancer screening at 5 centers from 2020 to 2022 were prospectively enrolled. Pancreas findings were designated as low-risk (fatty or chronic pancreatitis-like changes), intermediate-risk (neuroendocrine tumor [NET] <2 cm or branch-duct intraductal papillary mucinous neoplasm [IPMN]), or high-risk lesions (high-grade pancreatic intraepithelial neoplasia/dysplasia, main-duct IPMN, NET >2 cm, or pancreatic cancer). Harms from screening included adverse events during screening or undergoing low-yield pancreatic surgery. Annual screening was performed using endoscopic ultrasound and or magnetic resonance cholangiopancreatography. Annual screening for new-onset diabetes using fasting blood sugar was also performed ( ClinicalTrials.gov : NCT05006131). RESULTS: During the study period, 252 patients underwent pancreatic cancer screening. Mean age was 59.9 years, 69% were female, and 79.4% were White. Common indications were BRCA 1/2 (36.9%), familial pancreatic cancer syndrome kindred (31.7%), ataxia telangiectasia mutated (3.5%), Lynch syndrome (6.7%), Peutz-Jeghers (4.3%), and familial atypical multiple mole melanoma (3.5%). Low-risk lesions were noted in 23.4% and intermediate-risk lesions in 31.7%, almost all of which were branch-duct IPMN without worrisome features. High-risk lesions were noted in 2 patients (0.8%), who were diagnosed with pancreas cancer at stages T2N1M0 and T2N1M1. Prediabetes was noted in 18.2% and new-onset diabetes in 1.7%. Abnormal fasting blood sugar was not associated with pancreatic lesions. There were no adverse events from screening tests, and no patient underwent low-yield pancreatic surgery. DISCUSSION: Pancreatic cancer screening detected high-risk lesions with lower frequency than previously reported. No harms from screening were noted.

Reconstruction of Complex Oro-Mandibular Defect Using Free Fibula Flap and Lateral Sural Artery Flap: A Case Series.

Through and through complex oro-mandibular defect usually involves buccal mucosa, mandibular segment, lip, and outer cheek skin. Reconstruction of such extensive three-dimensional defects pose a great challenge to reconstructive surgeons which requires use of two flaps. There are diverse options for such types of defects like use two pedicled flaps, one free flap, one pedicled flap or use of two free flaps. Amongst them the use of dual free flaps is ideal for the reconstruction. Commonly used dual free flaps are free fibula osteocutaneous flap for mandible, buccal mucosal defect and free radial artery flap or antero-lateral flap for cheek defect. The major disadvantages of using these two free flaps include two different sites for flap harvest, more time for harvesting and increased overall surgery time. We present our experience of reconstruction of large oro-mandibular defect using free osteo-cutaneous fibula flap and lateral sural artery free flap from single limb in six patients between January 2019 and December 2020. Minimum follow up was 6 months.

Molecular characterization, putative structure and function, and expression profile of OAS1 gene in the endometrium of goats (Capra hircus).

An interferon-inducible gene, 2'-5'-oligoadenylate synthetase-1 (OAS1), plays an essential role in uterine receptivity and conceptus development by controlling cell growth and differentiation in addition to anti-viral activities. As OAS1 gene has not yet been studied in caprine (cp), so present study was designed with the aim to amplify, sequence, characterize and in-silico analyze the coding sequence of the cpOAS1. Further, expression profile of cpOAS1 was performed by quantitative real-time PCR and western blot in the endometrium of pregnant and cyclic does. An 890 bp fragment of the cpOAS1 was amplified and sequenced. Nucleotide and deduced amino acid sequences revealed 99.6-72.3% identities with that of ruminants and non-ruminants. A constructed phylogenetic tree revealed that Ovis aries and Capra hircus differ from large ungulates. Various post-translational modifications (PTMs), 21 phosphorylation, two sumoylation, eight cysteines and 14 immunogenic sites were found in the cpOAS1. The domain, OAS1_C, is found in the cpOAS1 which carries anti-viral enzymatic activity, cell growth, and differentiation. Among the interacted proteins with cpOAS1, Mx1 and ISG17 well-known proteins are found that have anti-viral activity and play an important role during early pregnancy in ruminants. CpOAS1 protein (42/46 kDa and/or 69/71 kDa) was detected in the endometrium of pregnant and cyclic does. Both cpOAS1 mRNA and protein were expressed maximally (P<0.05) in the endometrium during pregnancy as compared to cyclic does. In conclusion, the cpOAS1 sequence is almost similar in structure and probably in function also to other species along with its higher expression during early pregnancy.

Endoscopic management of hemorrhagic pancreatic fluid collections: A propensity-matched analysis.

Objectives: Hemorrhagic pancreatic fluid collections (hPFC) are a complication of pancreatitis with an unknown influence on prognosis. Advancements in endoscopic management of PFC have improved results over their surgical and percutaneous alternatives. We performed a propensity-matched analysis comparing clinical outcomes in hemorrhagic and non-hemorrhagic PFC (nhPFC). Methods: From November 2015 to November 2021, a retrospective comparative cohort analysis was performed comparing clinical outcomes for patients with hPFC and nhPFC managed with lumen-apposing metal stents. Propensity score matching was used to balance the two subgroups. Wilcoxon two-sample tests were used to compare continuous variables and Fisher's exact test was used to compare categorical variables. Kaplan-Meier method was used to estimate overall survival. Results: Fifteen patients with hPFC were matched with 30 nhPFC patients. Technical and clinical success was similar in both groups. The median length of hospitalization was 6 days in the hPFC group and 3 days in the nhPFC group (p = 0.23); however, more hPFC patients required intensive care unit admission post-procedure (33.3% vs. 16.7%, p = 0.26). Patients with hPFC were more likely to be readmitted to the hospital within 30 days (33.3% vs. 6.7%, p = 0.032). Mortality at 3 months (13% vs 3%, p = 0.25) and 6 months (27% vs. 7%, p = 0.09) was higher in the hPFC cohort. The 1-year survival estimate was 73.3% (standard error = 11.4) in the hPFC group and 88.9% (6.1) in the nhPFC group (p = 0.16). Conclusions: Patients with hPFC are more likely to be readmitted to the hospital within 30 days and have worse clinical outcomes.

Advances in Imaging of Inflammation, Fibrosis, and Cancer in the Gastrointestinal Tract.

Gastrointestinal disease is prevalent and broad, manifesting itself in a variety of ways, including inflammation, fibrosis, infection, and cancer. However, historically, diagnostic technologies have exhibited limitations, especially with regard to diagnostic uncertainty. Despite development of newly emerging technologies such as optoacoustic imaging, many recent advancements have focused on improving upon pre-existing modalities such as ultrasound, computed tomography, magnetic resonance imaging, and endoscopy. These advancements include utilization of machine learning models, biomarkers, new technological applications such as diffusion weighted imaging, and new techniques such as transrectal ultrasound. This review discusses assessment of disease processes using imaging strategies for the detection and monitoring of inflammation, fibrosis, and cancer in the context of gastrointestinal disease. Specifically, we include ulcerative colitis, Crohn's disease, diverticulitis, celiac disease, graft vs. host disease, intestinal fibrosis, colorectal stricture, gastric cancer, and colorectal cancer. We address some of the most recent and promising advancements for improvement of gastrointestinal imaging, including unique discussions of such advancements with regard to imaging of fibrosis and differentiation between similar disease processes.

Does Degree of Nasal Septal Deviation Measured with CT PNS have a Role in Management of Deviated Nasal Septum?

Deviated nasal septum is one of the most commonly encountered nasal anatomical abnormalities. Various studies showed that deviated nasal septum results in the osteo-meatal complex disease and compensatory turbinate hypertrophy. But only a few studies described the association between type and degree of angle of deviated nasal septum and measured the angle of septal deviation and explore its effect in causation of nasal symptomatology. Hence, this study was undertaken with the aim to study the relation between type and degree of septal deviation with nasal symptomatology and to measure the degree of septal deviation. This was an observational study and data collection was done from August 2017 to August 2019. A total 52 patients with symptomatic deviated nasal septum were studied. Higher septal deviation caused symptoms in 73% of cases and severe degree of angle of septal deviation caused symptoms in 63% of cases. High and severe deviated nasal are more associated with nasal symptoms. Therefore, we could derive a correlation between the cases with higher septal deviation and incidence of nasal symptoms.

Invasive fungal sinusitis-A neglected but important association with COVID 19 infection and a ticking bomb.

A retrospective analysis of severe acute respiratory syndrome (SARS) and influenza data worldwide has concluded that fungal coinfections associated with global SARS-coronavirus 2 (CoV-2) are likely to be missed or misdiagnosed. The coronavirus disease-2019 (COVID)-19 patients, especially those who are severely ill or immunocompromised, are more likely to suffer from invasive mycoses which require early detection and treatment. We report two such cases, one of which is a case of aspergillosis of unilateral orbit and maxilla and another case is of mucormycosis infection of the paranasal sinuses and bilateral orbits.

Two unusual conjugated fatty acids, parinaric acid and α-eleostearic acid, are present in several Impatiens species, but not in congener Hydrocera triflora.

Parinaric and α-eleostearic acids are unusual conjugated fatty acids. Unusual fatty acids, in general, are known to have roles in defense response; however, the role of parinaric acid in I. balsamina is not known, nor is it known whether it occurs in different species of Impatiens or its closest monotypic relative, Hydrocera triflora (L.) Wight & Arn. The aim of the study was to (a) characterize the fatty acid composition of 21 species of Impatiens and H. triflora and (b) determine whether parinaric and α-eleostearic acids are present in these taxa and, if so, (c) whether there is interspecific and intraspecific variation in parinaric acid content. Fatty acid profiling was done using gas chromatography and mass spectrometry (GC-MS). To uncover taxonomic patterns of variation in fatty acids, principal component analysis and hierarchical cluster analysis were performed. The major fatty acids in Impatiens were found to be palmitic (5.57-20.85%), stearic (2.86-21.61%), oleic (2.79-28.99%), linoleic (C18:2Δ9,12, 2.04-26.64%), α-linolenic (C18:3∆9,12,15; 11.07-53.99%), and four forms of parinaric acid (5.93-70.21%). Genus Impatiens contains two unusual conjugated fatty acids- parinaric and α-eleostearic, however these are absent in closely related H. triflora. This study reports the presence of four different forms of parinaric acid in Impatiens for the first time. Some species (I. mengtszeana, I. racemulosa and I. oppositifolia) were found to contain very high levels (> 50%) of parinaric acid and they might be useful for various biomedical and industrial applications. Apparently, the presence of parinaric acid is a characteristic of Impatiens. Significant variations were found in the amount and forms of parinaric acid. We propose the potential application of parinaric acid and α-eleostearic acid as chemotaxonomic markers for Impatiens. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01194-4.

Phosphatidic acid binds to and stimulates the activity of ARGAH2 from Arabidopsis.

Phosphatidic acid (PA) has emerged as an important lipid signal during abiotic and biotic stress conditions such as drought, salinity, freezing, nutrient starvation, wounding and microbial elicitation. PA acts during stress responses primarily via binding and translocating target proteins or through modulating their activity. Owing to the importance of PA during stress signaling and developmental stages, it is imperative to identify PA interacting proteins and decipher their specific roles. In the present study, we have identified PA binding proteins from the leaves of Arabidopsis thaliana. Mass spectroscopy analysis led to the identification of 21 PA binding proteins with known roles in various cellular processes. One of the PA-binding proteins identified during this study, AtARGAH2, was further studied to unravel the role of PA interaction. Recombinant AtARGAH2 binding with immobilized PA on a solid support validated PA-AtARGAH2 binding invitro. PA binding to AtARGAH2 leads to the enhancement of arginase enzymatic activity in a dose dependent manner. Enzyme kinetics of recombinant AtARGAH2 demonstrated a lower Km value in presence of PA, suggesting role of PA in efficient enzyme-substrate binding. This simple approach could systematically be applied to perform an inclusive study on lipid binding proteins to elucidate their role in physiology of plants.

Long-term transmural drainage of pancreatic fluid collections with double pigtail stents following lumen-apposing metal stent placement improves recurrence-free survival in disconnected pancreatic duct syndrome.

OBJECTIVES: Disconnected pancreatic duct syndrome (DPDS) is the most common cause of pancreatic fluid collection (PFC) recurrence. While long-term transmural drainage with plastic stents is the preferred endoscopic approach, there is a paucity of literature on patients undergoing initial drainage with lumen-apposing metal stents (LAMS). We describe our experience managing patients with DPDS. METHODS: A retrospective review of a prospectively maintained database (November 2015-September 2020) was performed looking at clinical outcomes and overall survival for patients undergoing endoscopic management of PFCs using LAMS. The primary outcome was to assess recurrence-free survival in PFC patients with DPDS managed with or without double pigtail stents (DPS) replacement after LAMS removal. RESULTS: Of 96 patients with PFCs, 48 with DPDS were included in the study. The median follow-up was 20.1 months. LAMS replacement with DPS was successful in 21/48 (43.8%) patients. Recurrence was seen in 1/21 (5%) patients with DPS replacement and 10/27 (37%) without DPS replacement. In multivariable models, a longer duration of LAMS placement was negatively associated with successful DPS replacement (odds ratio 1.33, 95% confidence interval [CI] 1.11, 1.59, P = 0.0019) and successful LAMS replacement with DPS in patients with DPDS improved recurrence-free survival (hazard ratio 0.09, 95% CI 0.01, 0.83, P = 0.033). CONCLUSION: In patients with PFCs and DPDS, early replacement of LAMS with DPS improves the likelihood of successful long-term transmural drainage and decreases recurrences.

Novel red colour emitting Ca0.995 Mg2 (SO4 )3 :0.5Eu2+ phosphor under ultraviolet, blue, and green excitation for plant growth LEDs.

In the recent few years, Eu2+ - and Mn4+ -activated phosphors are widely used as potential colour converters for indoor plant cultivation lighting application due to their marvellous luminescence characteristics as well as low cost. In this investigation, we synthesized novel red colour-emitting Ca(2-x) Mg2 (SO4 )3 :xmol% Eu2+ (x = 0-1.0 mol%) phosphors via a solid-state reaction method in a reducing atmosphere. The photoluminescence (PL) excitation spectra of synthesized phosphors exhibited a broad excitation band with three excitation bands peaking at 349 nm, 494 nm, and 554 nm. Under these excitations, emission spectra exhibited a broad band in the red colour region at ~634 nm. The PL emission intensity was measured for different concentrations of Eu2+ . The maximum Eu2+ doping concentration in the Ca2 Mg2 (SO4 )3 host was observed for 0.5 mol%. According to Dexter theory, it was determined that dipole-dipole interaction was responsible for the concentration quenching. The luminous red colour emission of the sample was confirmed using Commission international de l'eclairage colour coordinates. The results of PL excitation and emission spectra of the prepared phosphors were well matched with excitation and emission wavelengths of phytochrome PR . Therefore, from the entire investigation and obtained results it was concluded that the synthesized Ca0.995 Mg2 (SO4 )3 :0.5mol%Eu2+ phosphor has huge potential for plant cultivation application.

Differences in patient outcomes after outpatient GI endoscopy across settings: a statewide matched cohort study.

BACKGROUND AND AIMS: Outpatient GI endoscopy has been shifting from hospital outpatient departments (HOPDs) to ambulatory surgery centers (ASCs) in recent years. However, evidence on whether patient outcomes after endoscopic procedures are comparable across settings is limited. This study compares the incidence of unplanned hospital visits after GI endoscopy performed in ASCs versus HOPDs. METHODS: We conducted a retrospective cohort study examining unplanned hospital visits after outpatient GI endoscopy performed in Massachusetts during 2014 to 2017 using Massachusetts All-Payer Claims Database and Medicare fee-for-service claims. We identified screening colonoscopy, nonscreening colonoscopy, and esophagogastroduodenoscopies (EGDs) performed in ASCs or HOPDs and estimated unplanned hospital visit rates within 7 and 30 days after these procedures. To compare rates between ASCs and HOPDs, we constructed procedure-specific, propensity score-matched samples and used multilevel logistic regressions adjusting for patient, procedure, and facility characteristics. RESULTS: Seven-day unplanned hospital visit rates were 10.6, 18.3, and 38.9 per 1000 procedures for screening colonoscopy, nonscreening colonoscopy, and EGD, respectively, with significant variation across facilities. ASC patients consistently had fewer postprocedure hospital encounters. The relative risk of having 7-day hospital visits after screening colonoscopy performed in ASCs was .88 (95% confidence interval [CI], .79-.98) compared with HOPDs. The estimates were .84 (95% CI, .75-.94) for nonscreening colonoscopy and .57 (95% CI, .50-.65) for EGD. Thirty-day visits showed similar patterns. CONCLUSIONS: Unplanned hospital visits after outpatient GI endoscopy were not uncommon. However, ASC patients consistently had less frequent hospital-based acute care encounters, indicating that GI endoscopy could be performed safely in ASCs for select patients.

Expression of kisspeptin and its receptor in different functional classes of ovarian follicle in the buffalo (Bubalus bubalis).

Recently, we reported the differential expression of kisspeptinergic system in the bubaline hypothalamus and corpus luteum. Here, we document the expression of kisspeptin (Kp) and its receptor (Kiss1r) in the ovarian follicles of the buffalo with respect to the functional status. Follicles of ≥10 to ≤13 mm diameter (n = 45) were retrospectively categorized into active (n = 18), intermediate (n = 16) and atretic (n = 11) follicles based on the concentrations of intrafollicular progesterone (P4) and estradiol (E2). The P4:E2 ratio was significantly lower in the active follicle (0.43 ± 0.08) than that of the intermediate (3.46 ± 0.53) and atretic (28.4 ± 10.6) follicles (P < 0.05). Relative fold change in the transcripts of kisspeptin (Kiss1), Kiss1r, gonadotrophin receptors, steroid acute regulatory protein (StAR), cytochrome P450 family 11 subfamily A member 1 (CYP11A1), cytochrome P450 Family 19 subfamily A member 1 (CYP19A1), insulin like growth factor -1 (IGF-1), apoptotic factors (caspase 3 and B-cell lymphoma 2, BCL2) was calculated using qPCR in the follicular wall of the three categories of follicle (n = 8/group). In another experiment, histological sections of the ovary (n = 41) were used to group the follicles as described above and immunostaining of Kp, Kiss1r and aromatase was done. A significant upregulation of StAR, CYP11A1 and CYP19A1 in the active follicles supported the endocrine basis of follicular classification. The transcripts of Kiss1 and Kiss1r were upregulated by 19.45 fold and 4.25 fold, respectively in the active follicle as compared to other groups. Immunolocalization studies revealed that Kp and Kiss1r were localized to the basal and antral granulosa cells (GC) of the active and intermediate follicles; however, the staining intensity was stronger in the former group. Strong expression of CYP19A1 in the GC layer of active follicle supported the histological basis of defining the functional status of the follicle. It is concluded that the follicular compartment of the bubaline ovary expressed the constituents of kisspeptinergic system. The expression of Kp and Kiss1r was influenced by the functional status of the follicle with intense localization in the GC layer of the active follicles.

Functional characterization and expression profile of microsomal FAD2 and FAD3 genes involved in linoleic and α-linolenic acid production in Leucas cephalotes.

The genus Leucas belongs to Lamiaceae, and has attained more attention due to the presence of unusual allenic fatty acids called laballenic and phlomic acid in majority of its species. This genus has been known since traditional medicinal times and has numerous economical, nutritional, and industrial properties. So far genetic, molecular and biochemical analyses of lipid metabolism and fatty acid biosynthetic pathway in Leucas has not been reported. The objective of this study is to identify, isolate, analyze expression profiles, and functionally characterize the membrane-associated desaturases responsible for unsaturated fatty acid accumulation in Leucas cephalotes. Full-length LcFAD2 and LcFAD3 cDNAs were isolated and expressed in Saccharomyces cerevisiae BY4741 for functional characterization. Substrate feeding assay using S. cerevisiae confirmed that the LcFAD2 enzyme catalyzes desaturation of both palmitoleic (16:1∆9) and oleic (18:1∆9) acids to form palmitolinoleic (16:2∆9,12) and linoleic (18:2∆9,12) acids respectively. As a contrast, the heterologous activity of LcFAD2 enzyme in S. cerevisiae led to the synthesis of palmitolinoleic (16:2∆9,12) acid, an unusual fatty acid that is not found naturally in Leucas cephalotes. While the LcFAD3 enzyme catalyzed linoleic acid (18:2∆9,12) into α-linolenic acid (18:3∆9,12,15). Furthermore, transcript abundance of LcFAD2 and LcFAD3 cDNAs were estimated from various plant parts such as roots, shoots, leaves, petals and developing seeds. Our results have shown that the differential transcriptional activity of LcFAD2 and LcFAD3 desaturase genes differs significantly in developing seeds, petals, leaves, stems, and roots of L. cephalotes. Furthermore, for the industrial production of these essential fatty acids, namely, linoleic and α-linolenic acid, FAD2 and FAD3 enzyme activity could be exploited from this upcoming significant oil plant, Leucas cephalotes. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12298-021-01016-z.